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TRANSACTIONS

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Microscopical Society

Organized 1878

Incorporated 1891

PUBLISHED QUARTERLY

BY THE SOCIETY

EDITED BY THE SECRETARY

PAUL S. WELCH

ANN ARBOR, MICHIGAN

VOLUME XLI

Number One

Entered as Second-class Matter August 1^, 1918, at the Poit-offiie at Menasha

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special rate of postage provided for in Section 1 103, of the

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CiEOKOE Banta Publishing Company

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1922

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TABLE OF CONTENTS

For Volume XLI, Number 1, January, 1922

The External Morphology of Lachnosterna crassissima Blanch. (Scarabs idae,

Coleop.), with nine plates, by Wm. P. Hayes 1

The Respiratory Mechanism in Certain Aquatic Lepidoptera, with two plates, by Paul

S. Welch 29

Department of Methods, Reviews, Abstracts, and Briefer Articles

Dichromatic Illumination for the Microscope, with two figures, by L. A. Hausman . 51

A Modified Barber Pipette, with one figure, by Bert Cunningham 55

Cleaning Slides and Covers for Dark-field Work, by S. H. Gage 56

Proceedings of the American Microscopical Society 57

TRANSACTIONS

OF

American Microscopical Society

(Published in Quarterly Instalments)

Vol. XLI JANUARY, 1922 No. 1

THE EXTERNAL MORPHOLOGY OF LACHNOSTERNA

CRASSISSIMA BLANCH

(Scarabseidae, Coleop.) '

By

Wm. p. Hayes

Assistant Entomologist, Kansas Agricultural Experiment Station

Introduction

The present discussion of the external features of Lachnosterna craisis-

sima Blanch, is offered to supply the lack of an available treatise in English

on the morphology of the beetles belonging to the family Scarabaeidae.

The nearest approach to the subject is the famous historical French work

of Straus-Durckheim "Considerations Generales sur L'Anatomie Comparee

des Animaux Articules" in which is included a description and many fine

drawings of the anatomy of Melolontha vulgaris, Hanneton. This work

appeared in 1828 and is a masterpiece of its kind. However, much of the

anatomical nomenclature is now antiquated and the work itself hard to

secure, consequently the present paper, dealing with a species of a closely

allied genus of the same family, is here presented.

The nomenclature used by the writer is that which was deemed most

useful. It follows no one system of the modern writers but is adopted from

such writers as Snodgrass, Crampton, and others.

This species of the genus Lachnosterna was chosen for study because of

its abundance in the vicinity of Manhattan, Kansas. Of the 23 species

found in this vicinity, L. crassissima ranks first in relative numbers. A

total of 86945 specimens of this genus have been collected by Mr. J. W.

' Contribution No. 54 from the Entomological Laboratory, Kansas State Agricultural Col-

lege. This paper embodies the results of some of the investigations undertaken by the writer

in the prosecution of project No. 100 of the Kansas Agricultural Experiment Station. The

writer wishes to acknowledge his indebtedness to Dr. P. S. Welch for many helpful criticisms

during the course of preparation of this paper and to the American Microscopical Society for a

grant from the Spencer-Tolles Fund to publish the accompanying drawings.

1

2 \VM. P. HAYES

McColloch and the writer during the years 1916-1920, and of this number

30230 were L. crassissima. The specimens were preserved in 70% alcohol,

boiled in potassium hydroxide when ready to use, and studied under the

binocular. ^ „

General Considerations

Size. — The adults of this species are among the most broadly ovate of

the genus Lachnostcrna. They vary greatly in length, width, and maximum

depth, and, on an average, the females are somewhat longer than the

males, as well as much broader posteriorly. The greatest body depth varies

in individuals of both sexes. Not only in size is this difference noticeable,

but also in respect to the regions of the body in the two sexes. The ma^es

have their region of greatest depth through the thorax, which was found

to average 7.1 mm., while the females are deepest through the posterior

end of the abdomen where they average 7 .6 mm. Some of the depth varia-

tion in the females is due, in part, to the degree of flexibility of the inter-

segmental grooves, especially when the abdomen is distended with eggs.

Because of the deflexity of both the labrum and pygidium, the maximum

length of the males and females was arbitrarily measured from the emargi-

nation of the clypeus to the basal or proximal edge of the pygidium and not

to the extreme ends as is the usual practice. The average length of 25

males, chosen at random, was 18.2 mm. and, for the same number of

females, 18.7 mm. The maximum length of the males was 20 mm. and

of the females 20.9 mm., while the minimum was 16.3 mm. for the males

and 16.5 mm. for the females. Width measurements were made at

seven different regions of the body to get a general notion of the width

variation. These regions were chosen arbitrarily as follows:

1 — At anterior margin of the eyes

2 — At anterior margin of prothorax

3 — At lateral angles of prothorax

4 — At base of prothorax

5 — At base of elytra

6 — At bulge near middle of elytra (widest points)

7 — At declivity of elytra near the distal end.

The table on the opposite page shows the average width, length, and

depth measurements of 25 males and 25 females.

In individual specimens, the length measurement is highly variable,

depending on the character of the specimens at hand. Alcoholic and living

specimens are extensile because of the telescopic nature of the union of

head and thorax and to a lesser degree the thorax and abdomen, thus

causing differences in length. Dried specimens will, on the contrary, i)er-

mit of more constant measurements.

Color. — The general mass color of this species is chestnut-brown or

castaneous (dragon's blood plus a slight admixture of vermilion Smith's

EXTERNAL MORPHOLOGY OF LACHNOSTERNA CRASSISSIMA

color chart, 1906, p. 154). However, some specimens vary to a dark

brown, almost black, in which case a grayish iridescence is vjery apparent.

Dorsally, the head, thorax, and elytra are shining and in certain lights a

gray color exists. This grayish tinge is a structural color caused by fine

striae on the elytra. These striae are also present on the thorax but the

iridescence is not so pronounced. The eyes are black and prominent.

Ventrally the ground color is castaneous. The thorax is covered with dull,

yellow hairs, 1.5-2 mm. long, somewhat sparsely scattered on the pro-

thorax, but several times denser on the meso- and metathorax. The abdo-

men frequently has a grayish super-color imparted by an adhering exuda-

tion which can be scraped off. The legs and antennae are lighter brown,

almost ferruginous in color.

Table I. — Average Size of Various Body Regions

Skxi'al Dimorphism

}falc. — The clul) of the antennae (Plate I, tig. 4) is about equal in

length to the scape (s.) and funicle (fu.) combined. At the middle of the

ventral surface the al:)domen is longitudinally depressed and the penulti-

mate ventral segment bears a faint transverse carina near its distal end.

Frequently, this carina is nothing more than a slightly wrinkled convexity.

Immediately behind this carina the last, or ultimate, ventral segment has

a deep, rounded fovea whose distal margin is obtusely and angulately

emarginate. On the posterior tibia, the inner spur is about one-half the

length of the outer and somewhat more slender spur. The pygidium is

not gibbous and is more noticeably truncate at the distal end than is this

structure in the female. The hind tarsi are longer than those of the female.

Female. — The club of the antenna (Plate I, Fig. 5) is about as long as

the funicle. The ultimate ventral abdominal segment is somewhat broadly

and rather deeply emarginate distally, and the fovea is absent. The pygi-

dium is gibbous, smooth, and shining at the apex of the gibbosity and more

4 WM. p. HAYES

exposed than in the male. Ventrally, the abdomen is more broadly rounded

and shining than that of the male and the longitudinal depression is lacking.

The inner spur of the hind tibia is about equal in length to the outer and

about as wide as the same spur in the males, while the hind tarsi are

shorter than in the male. The tooth of the tarsal claws is somewhat larger

than this tooth in the male.

Contrast of Body Surfaces

Dorsal Surface. — The dorsum of the head, thorax, and abdomen, by a

casual examination, appears smooth and shining, but closer scrutiny reveals

minute punctures over the entire surface. These punctures are more

numerous on the front than on any other part of the body. Here they are

closely placed, and often confluent. The clypeus is less densely punctured

and the punctures are about equal in density to those on the thorax. A

setigerous canthus is found on the eye. On the thorax a faint, smooth,

median, longitudinal line is formed by the absence of punctures. Laterally,

the punctures are less dense. The lateral margin is serrate and hirsute, and

at the base transverse channels extend mesad, failing to reach the median

line. The scutellum is large, somewhat heart-shaped, irregularly and less

densely punctured. The elytra are likewise irregularly punctured, and

five indistinct costse occur on each elytron.

Ventral Surface. — Ventrally, the head is dark-brown, sparsely hairy

and, in part, concealed under the anterior ventral margin of the thorax.

The thorax is thickly covered with pale, yellow liairs about 1 .5 mm. long,

and the legs are sparsely covered with shorter hairs. The punctures of

the ventral abdominal surface are, on the whole, smaller than the dorsal

ones. Each bears a short, recumbent hair and is more widely separated.

There are about the same number of punctures per square millimeter as

on the upper surface where they are larger l)ut more closely placed.

Contour

The transverse contour of the body exhibits four distinct geometrical

figures in three principal regions of the body. Through the head a narrow,

elongated oval is apparent, through the thorax a broad oval, while the

abdomen shows a somewhat different contour for each sex. In the female,

it is broadly oval, almost circular, and in the male much the same, except

for the ventral surface where the oval is .somewhat flattened, due to the

fovea OP the lower surface of the abdomen.

Body Divisions

The three general regions of the body are to be recognized by definite

sutures which separate them. The head is the smallest division, being

less than one-half the width of the thorax. It is greatly depressed and

deflected. From above only the clypeus, front and eyes can be seen, as

EXTERNAL MORPHOLOGY OF LACHNOSTERNA CRASSISSIMA 5

the labrum is deflected on the under side of the head. The head is tele-

scoped within the thorax and connected to it by a sclerite-bearing cervicum

or the so-called microthorax.

The thorax is next in size to the head, somewhat oblong, being nearly

twice as wide as long. The sides are nearly parallel at the base, but con-

verge anteriorly. The oval contour of the three segments of the thorax is

nearly similar in each with perhaps a more flattened aspect of the pro-

thorax. Dorsally, the meso- and metathorax each bear a single pair of

wings, those of the mesothorax being modified to form the elytra which

cover the metathoracic wings. Each segment of the thorax bears a pair

of jointed legs.

The abdomen, which is larger than the other two divisions combined,

is completely covered by the elytra, except at the pygidium. The pygidium

is more exposed from above in the female than in the male. On the lower

surface are found only six abdominal segments, while eight are apparent

en the upper surface after removing the elytra. This condition is appar-

ently constant in both sexes. The sexual differences of the abdomen have

already been described. The abdomen is telescoped into the thorax on the

lower side, and united to the metacoxai. Dorsally, the connection is made

by a membrane to the postscutellum, and the parts do not overlap. The

intersegmental grooves on the dorsum are comparatively wide and the

fusion of the segments is loose and flexible, while ventrally, the segments

are closely fused, forming narrow, curved grooves. Seven pairs of spiracles

are to be found on the abdomen, only one of which can be seen below the

elytra. The others are mostly in the ridges formed in the pleura.

The Structure of the Head

Front and Vertex. — The size and general contour of the head (Plate I,

figs. 1-2-3) have already been noted. It is partly withdrawn into the

prothorax and the mouth parts are wholly on the inferior surface with but

a small part of the labrum visible from above. The front (Plate I, fig. 1,

fr.) is a large somewhat rectangular area lying between the eyes, and

limited anteriorly by the suture separating it from the clypeus (cly.).

Its outer angles are extended to form the canthus of the eye (en.). No

epicranial suture is present and the region is not sharply separated from the

vertex. The front is closely and strongly punctate. Near the vertex^ at

the point where the prothorax overlaps the head, the punctures disappear

rather abruptly, except near the eyes, leaving a strong line of demarkation

between the punctured area and the smoother region of the vertex and

occiput. A few scattered punctures are to be found in these regions. Each

puncture bears a recumbent hair which is inclined anteriorly. The vertex

(v.) merely consists of the upper region of the head having no definite

6 WM. p. HAVES

boundaries but lying between the front and the occiput; the occiput being

the posterior region of the head lying above the opening of the occipital

foramen. No ocelli are present.

The Clypeus and Canthus. — The clypeus (Plate I, fig. 1, cly.) is situated

on the anterior margin of the front, and the suture separating them, which

is strongly sinuate, is known as the clypeo-frontal suture. The clypeus

is somewhat rectangular, being twice as wide as long. Numerous punctures

are present but they are not as dense as on the front. The anterior margin

is slightly emarginate and a strong upturning gives to the whole sclerite

a deeply concave appearance. The postero-lateral corners of the sclerite

are bordered by the eyes and at this point a chitinous process protrudes

upon and partly divides the eyes. This process, known as the canthus

(Plate I, figs. 1 and 2, en.), appears somewhat as an extension of the

clypeus, but in reality is a continuation of the anterior corners of the

front lying partly under the clypeus. Hairs are scattered over the surface

of the canthus.

The Lahrum and Epi pharynx. — The labrum (Plates I and II, hg. 1,

labr.) is attached to the anterior border of the clypeus, being greatly

deflected and nearly hidden by the clypeus. Dorsally it is somewhat semi-

circular in outline and is depressed to form a deep fovea near the center.

It is covered with long thinly placed hairs. On its inner surface are two

convergent rows of mesading point hairs (Plate II, fig. 1).

The epipharynx (Plate II, fig. 1, epi.) is greatly reduced in this species.

It has almost disappeared because of the extension of the labrum over

most of its entire surface. A somewhat triangular elevated clump of

hairs, or spines, is the most conspicuous remnant of the epipharynx.

The Eyes. — The eyes (Plate I, figs. 1-2-3, e.) are the most prominent

part of the head. They are large, somewhat oval bodies on the dorsal,

lateral and ventral regions of the head. They are nearly divided dorsally

by the canthus.

The facets (Plate I, fig. 6, fac.) which are about .021 mm. in diameter,

average about vS80 to the square millimeter. In shape they are somewhat

regularly hexagonal and each hexagon is the cornea of a completely dis-

tinct eye.

Gena and Gula. — The la,teral parts of the cpicranium form the genae

(Plate I, figs. 2 and 3, g.) whose ventral limits are determined by the sutures

separating the genae from the large head sclerite — the gula (Plate I, figs.

2 and 3, gu.). The gula occupies about one-third of the ventral surface

of the head. It is somewhat quadrate in outline, being slightly wider

anteriorly where it is separated from the submentum by a transverse

suture. The lateral margins are limited by the gular sutures and posteriorly

by the cervical membrane.

EXTERNAL MORPHOLOGY OF LACHNOSTERXA CRASSISSIMA 7

The Occipital Foramen and Tentorium. — The occipital foramen, or

foramen magnum, is the large opening in the head, opposed to a like

opening in the thorax.

Through the occipital foramen can be seen, within the head, a chitinous

structure, the tentorium (Plate V, figs. 1-2-3, tent.). A large arch-like

structure represents the body of the tentorium, while a pair of small,

short, posterior arms (post, a.) are present. The anterior arms (ant. a.)

are broad structures extending cephalad from the body of the tentorium,

and the dorsal arms (dor. a.) are represented by a pair of short pointed

processes extending cephalad into the head cavity.

The Cervicum and its Sclerites. — The membranous area between the

head and thorax is known as the cervicum. It contains six cervical scler-

ites. No attempt to homologize these sclerites has been made. The draw-

ing (Plate V, fig. 4) shows the left side of the cervicum with the dorsal edge

to the right and the anterior edge toward the top. It will be seen that near

the dorso lateral margin is a small hair-bearing sclerite and near the ventro

lateral margin are two sclerites, a large anterior one which overlaps a

small posterior one.

The Head Appendages

The Antennce. — The antennas (Plate I, figs. 4 and 5) have been pre-

viously mentioned under the discussion of sexual differences. The male

(Plate I, fig. 4) has a much larger club than the female (Plate I, fig. 5).

Normally both sexes have 10 segments, three of which go to make up the

lamellate club (cl.). The others form the funicle (fu.) and scape (sc).

The Mandibles. — The mandibles (Plate III, figs. 1 and 6) are large

complicated structures bearing on their inner surface a large, oval, grinding,

or molar surface (mo.). Extending over the molar surface are a number

of transverse ridges which are used in the process of grinding food. The

anterior end of the mandible is thought to be the homologue of the galea

of the maxilla. It is modified into a sharp cutting edge with two blunt

teeth (gal.). Near the anterior end of the molar surface is a membrane

(memb.) bearing various shaped spines and setae which are shown in Plate

III, figure 5. Immediately caudad of the molar surface is still another

membrane (memb.) bearing short broad stiff spines. These are shown

enlarged in Plate III, figure 3. A transverse section through the molar at

the junction of the ridges (Plate III, fig. 4) shows a flat surface with small

ridges extended ectad over half the surface.

Two chitinous apodemes (Plate III, fig. 2) are attached to muscles

controlling the movement of the mandible.

The Maxillce. — Each maxilla (Plate II, fig. 3) is divided into five prin-

cipal regions: the cardo, stipes, palpifer, galea and lacinia. There seem

to be no sutures delimiting a subgalea or dividing the galea into two lobes.

The cardo (cd.) is rather short and broadly club shaped, being constricted

8 WM. p. HAYES

somewhat posteriorly. Across the center is an abrupt change in contour,

making the anterior region of the cardo much thicker dorso ventrally. This

change of level is represented in the drawing by the transverse dotted line.

The stipes (st.) is the large median triangular sclerite, alongside of which

is the long narrow palpifer (max. pf.) bearing a four-jointed palpus (p.).

On the margin of the stipes opposite the palpifer is a large somewhat

triangular area with one corner elongated to form a large spine-bearing

lobe. This is the lacinia (lac). On the ectal margin of the lacinia is a

large five-toothed heavily chitinized structure, the galea (gal.).

The Labium and Hypopharynx. — The labium (Plate II, fig. 2) is sep-

arated from the gula by a transverse suture which extends across the

ventral surface of the head in the region where the cardo of the mandible

is articulated. Following Kadic's (1902, pp. 207-228) interpretation of

the labium of Coleoptera, we find the following regions:

The submentum is divided transversely into two regions, the anterior

plate (Ap. Sm.) and the posterior plate (pp. sm.). The posterior plate is

attached to the gula and is much wider at the postero lateral margins,

somewhat constricted at the middle and slightly broader anteriorly. The

anterior plate is more nearly quadrate, broader than long, and with the

lateral edges rounding out to form a bulge near the middle of the sclerite.

The mentum is separated from the anterior plate of the submentum by a

transverse suture which has a distinct emargination near its center. Simi-

larly, it is somewhat broadly quadrate and bears a few mesad pointing

hairs. The anterior marein is strongly biemarginate.

The glossa and paraglossae (Plate IV, fig. 1) are not evident on the

ventral surface but are bent within the buccal cavity. The glossa (gl.)

is a single median sclerite, while the paraglossae (pig.) are found on either

side of it. They bear the three-jointed labial palpi. These structures are

not easily located without having well cleared specimens. Near the base

of the palpus on the inner surface is a diagonal suture limiting an area

termed the squama palpigera (sq. pi.).

The hypopharynx (Plate IV, fig. 1-3 hyp.) is a V-shaped spiny struc-

ture lying on a clump of spines or strong hairs, principally on the inner

surface of the anterior plate of the submentum. Caudad and dorsad to

the hypopharynx are two long, narrow, chitinous structures known as the

fulcrum hypopharyngeum (ful. hyp.). At the dorso-posterior end are two

small transverse sclerites constricted somewhat near their middle. These

are the pharyngeal sclerites (phy. scl.). To these structures are attached

the anterior margins of the pharynx (phar.) and just posterior to the

hypopharyngeum, on each side, is a row of backward pointing hairs. An-

teriorly, the two arms of the fulcrum hypopharyngeum unite under the

pharynx to form a sort of strengthening apparatus for the spiny structure

underlying the hypopharynx which extends forward to form three arms.

EXTERNAL MORPHOLOGY OF LACHNOSTERNA CRASSISSIMA 9

These are shown on their ventral aspect in Plate IV, figure 2. A lateral

view showing the relation of these parts to the pharynx (phar.) is shown

in Plate III, figure 3.

The Prothorax

Protergum. — (Plate VI, fig. 1). The tergum of the prothorax or pro-

notuni is convex, nearly twice as broad as long, with the sides somewhat

narrowing from the base to the apical margin and constricting rather sud-

denly anteriorly. The lateral margins are distinctly crenate and ciliate,

but not so represented in the drawing. A deep emargination occurs on

the anterior margin which overlaps the head, extending to the middle of

the eyes. The posterior margin is broadly angulate and overlaps, as far

as the elytra, the mesothorax to which it is connected by a membrane.

Close, though not very dense, punctures cover the surface, and a smooth

median caudo-cephalad line is faintly evident. On each side near the

posterior margin is an incipient channel extending from the postero-lateral

angle to within a short distance of the middle.

Pleura. — At the lateral margins, the tergum is not inflexed to form

the so-called prothoracic epipleurae which are strongly evident in some

Coleoptera (e.g. PterosUchus calif ornicus Dej.). The prosternum (Plate

VI, fig. 2, pro. ster.) and the pleural sclerites compose the ventral aspect

of the prothorax. The episternum (Plate VI, fig. 2, eps.) and the epimeron

(epm.) have no line of separation. Anteriorly, the episternum elongates

mesally to fuse with the sternum whose anterior margin turns inwardly

to form a phragma. Likewise, the epimeron extends mesally, tapering

towards its extremity. Thus the two extensions form the anterior and

posterior margins of the coxal cavities (cc). The junction of the epimeron

with the posterior region of the sternum creates in other Coleoptera the

closed coxa] cavities. These are partly open. This species has no suture,

as in Melolontha vulgaris, separating the sternum from the episternum.

Prosternum. — The prothoracic sternum (Plate VI, fig. 2, pro. ster.)

occupies the inferior, median region of the prothorax. It is quite irregular

in shape and has, as mentioned before, no distinct line of demarkation

separating it from the episternum. Externally, a noticeable feature is a

caudad-projecting tongue between the cavities of the coxae. At the anterior

end of this tongue an irregular, circular ridge causes the formation of a

somewhat rounded depression of the sternum. This tongue-like projection

after attaining the posterior margins of the coxae is expanded at right

angles and extends laterally to meet the epimeron of each side. Inter-

nally, after the removal of the coxae, the sternum will be found to have

enlarged into a somewhat rectangular piece with rounded postero-lateral

corners. It tends to form a concavity in which a part of each coxa rests.

On the sides of the anterior edge of the internal sternum are two prolonged

10 WM. P. HAYES

entosterna] apophyses (Plate VI, fig 3, es. aph.) which extend dorso-

laterally.

The first pair of spiracles located ventrally are suspended in the mem-

brane which unites the prothorax to the mesothorax.

The Prothoracic Legs

The Trochantin. — (Plate VI, fig. 6, tn.). The trochantin is a small

piece hidden within the interior of the prothorax, which, when viewed

from its caudal aspect, presents a depressed or cup-like structure articulat-

ing with the anterior margin of the coxa. The latero-dorsal margin bears

a small, somewhat sharpened corner that is loosely articulated with a small

apodeme on the inner surface of the prothoracic episternum. The end of

the coxa articulates with the lower end of this same apodeme.

The coxa. — (Plate VI, fig. 6, ex.). The coxa of the anterior leg is

cylindrical in form, and slightly over three time as long as its greatest

diameter. It lies transversely in the coxal cavity of the prosternum and

extends laterally under the edges of the pleura, thereby concealing the

articulation with the trochantin. On the inner surface is a large opening

extending from near its lateral extremity to nearly half its length.

The cephalic edge of this opening articulates with the trochantin and the

caudal edge is connected by a membrane to the arms of the epimeron lying

immediately behind. The opening is partly closed by an overlapping

of its edges which serve as places of attachment for several muscles.

At the distal end the coxa likewise articulates with the sternum near

the mid-ventral line, and is thus fixed at both ends so that it moves in a

rotary manner on its axis. There is a second opening at the distal end which

receives a prolongation of the trochanter and thus permits of articulation

at this point.

yiie Trochanter. — (Plate VI, fig. 6, tr.). The trochanter is a small

triangular piece lying between the co.xa and femur, articulating with both

the coxa and femur, being more firmly attached to the latter.

The Femur. — (Plate VI, fig. 6, f.). The femur is about as long as the

coxa, is somewhat flattened and bears on its inner surface a groove-like

depression in which the tibia may rest when folded back on the femur.

A socket is located in its distal end which receives a condyle from the tibia,

forming a ball and socket articulation ])ctween the femur and tibia.

The Tibia. — (Plate VI, fig. 6, t.). The tibia is remarkal)ly adapted for

burrowing in the soil. It is somewhat ol)lifiuely truncate at ils aj^ex,

about equal in length to the femur, and is strongly compressed, esj)ccially

at its anterior edge wliich bears the three tibial teeth. Of these the ter-

minal tooth is very strong and about as long as the first tarsal joint, while

the other two are broader and not so long. Near the femur the tibia is

rather cylindrical and bears a terminal condyle for articulation with the

EXTERNAL MORPHOLOGY OF LACHNOSTERNA CRASSISSIMA 11

femur. On its external margin, opposite the three teeth, is a strong mova-

able spine.

The Tarsus. — (Plate VI, fig. 6, tar.). The tarsus is composed of five

segments. The first four are about of equal length, the fifth slightly

longer. They are cylindrical, and become enlarged distally. The terminal

segment bears a pair of large claws (t. cl.) each of which bears an intra-

median (male) or median (female) tooth. The tooth is slightly larger in

the female.

The Mesothorax

The Mesotergum. — (Plate VII, figs. 1, 2, 3). The mesothorax is the

smallest of the three divisions of the thorax; it bears the second pair of

legs and the first pair of wings modified to form the elytra (Plate VI, fig. 4).

The dorsal or tergal region of the mesothoracic segment is occupied largely

by the somewhat triangularly shaped scutellum adjoined to which are the

points of attachment of the elytra (Plate VI, fig. 5). Three regions of the

four which comprise the typical thoracic tergum (Snodgrass, 1909, p. 523)

are to be distinguished in this species, namely, the praescutum, scutum and

scutellum. The postscutcllum, or pseudonotum of some writers is absent.

The Praescutum (Plate VII, figs. 1 and 2, praes. ph.) is composed of a

small meso-cephalad projecting phragma between the prothorax and the

metathorax. It is slightly concave and is reinforced at its cephalic margin

by a somewhat heavier deposition of chitin, which forms a rod-like brace

extending from one edge of the praescutum to the other (Plate VII, fig. 3).

The scutellum (Plate VII, figs. 1, 2, 3, scl.) is a large somewhat triangularly

shaped sclerite part of which is covered by the elytra leaving the posterior

half exposed externally. Its anterior or covered portion is somewhat

closely punctate and covered with recumbent hairs while the exposed

part is sparsely punctate, devoid of hairs and rather shining. The lateral

margins each bear, on their anterior halves, two mesad-projecting pieces

which represent the divided regions of the scutum (Plate VII, fig. 2, set.).

These phragma-like pieces are nearly triangular in shape and each ter-

minates in a pointed ventro-cephalad projecting process which rests on

the underlying metathoracic praescutal phragma (Plate VII, fig. 2, prs. ph.),

and articulates with its antero-lateral projecting corners. At the cephalic

margin of the scutum is a small depression or cavity in which the third

axillary of the elytra lies when in the state of rest. The caudal margins are

extended backward and unite to form a semicircular redupUcation on the

inferior surface of the exposed port'on of the scutellum (Plate VII, fig. 3,

scl. red.). To this redupUcation is attached a small membrane which

connects the mesothoracic tergum to the lateral and caudal margins of the

metathoracic praescutal phragma. The cephalic margin of the scutellum

bears a membrane which connects the prothorax and mesothorax. The

12 WM. p. HAYES

cephalo-lateral angles of the scutellum articulate with a small, sharp

process which projects ventrad and unites with the anterior margin of the

mesoepisternum.

The Elytra. — (Plate VI, figs. 4 and 5). The elytra which cover the

meso- and metathorax and the greater portion of the abdomen are large,

somewhat rectangular wing covers extending caudad to the middle of the

penultimate abdominal segment, leaving the pygidium exposed. Their

lateral and posterior margins are somewhat abruptly declivitous. The

upper surface bears faint traces of the nervures and at each humeral angle

there is a slight protuberance. The elytra are inserted on the mesothorax

between the scutellum and the mesopleura. The base of the wing covers

is somewhat truncated and curves ventrad. Near the middle of the basal

margins on each elytra is a strong, bifurcated apophysis (Plate VI, fig. 5),

which articulates with the wing process of the mesothorax, there are three

principal wing axillaries (Plate VI, fig. 5, 1 ax., 2 ax., 3 ax.) in the membrane

which are very irregular in shape and impart a different appearance from

every aspect in which they are viewed.

The interlocking mechanism of the elytra is similar to that described

for Lachnosterna fusca by Breed and Ball (1908, p. 291) who found in

Coleoptera four devices for fastening the elytra in place. These are

described by these writers as follows:

1. By a co-adaptation of the elytra along the dorsal suture.

2. By means of a groove on the dorsal face of the metathorax into

which the swollen inner edges of the elytra fit.

3. By slipping the anterior edges of the elytra under the scutellum

and hooking them (a) on to the scutellum, or (b) on to the metathorax.

Pressure derived from the retracted prothorax may aid in keeping these

edges n position.

4. By hooking the anterior lateral edges of the elytra over ridges or

into grooves on the lateral faces of the metathorax.

In Lachnosterna, the first three methods are used to interlock the elytra

while the fourth is present but not functional.

The Mesopleura. — (Plate VII, figs. 4 and 5). The mesopleuron consists

externally of two sclerites, the episternum (eps.) and the epimeron (epm.).

The episternum is a subrectangular plate with a strongly rounded dorsal

margin, which adjoins the alar membranes. The anterior and posterior

margins are nearly parallel, the former serving as a place of attachment

for the intersegmental membrane, and the latter bordering on the epimeron.

The ventral margin is attached to the mesosternum (ms. ster.). The epi-

meron (epm.) is nearly trapezoidal in shape with the cephalic and caudal

margins nearly parallel. The cephalic margin connects with the epister-

num, the caudal one joins the metathorax, the dorsal margin gives attach-

ment to the alar membrane and the ventral margin tapers to meet and

EXTERNAL MORPHOLOGY OF LACHNOSTERNA CRASSISSIMA 13

connect with the coxa of the mesothoracic leg (ex.). Between the coxa

and the episternum is a small, narrow sclerite not visible externally — the

trochantin (Plate VII, fig. 4, tn.) which articulates by means of a small

condyle with the coxa. This sclerite is not present in the corresponding

region of the metathorax.

An internal view of either of the mesopleura (Plate VII, fig. 5) shows a

strong entopleural structure arising along the suture separating the epis-

ternum and epimeron and forming a pleural ridge which tapers at its

ventro-mesal angle into a pleural arm (pi. a.) extending into the body

cavity and terminating in a cup-shaped disk which serves for the attach-

ment of muscles. The caudal margin of the epimeron presents internally

a strong reduplication which aids in concealing the spiracles of the second

pair of respiratory organs. The spiracles are not visible externally but lie

in the suture between the mesoepimeron and the metaepisternum. The

dorsal margin of the episternum is modified into a strongly chitinized

blunt process, constituting the wing process on which articulates the

bifurcated apophysis of the elytron.

The Mesosternum — (Plate VII, figs. 6 and 7, ms. ster.). The mesoster-

num is a transverse quadrilateral plate whose anterior margin serves as

a place of attachment for the intersegmental membrane lying between the

pro- and meso thorax. Its lateral edges border on the ventral margin of

the mesoepisternum, and the posterior margin presents a biemarginate

appearance with a median, caudal projecting piece which extends between

the coxae. The whole external posterior margin is bordered by the meso-

coxal cavities. With the coxa removed, it can be observed that the pos-

terior margin is extended into a concave process in each cavity in which the

coxa lies at rest. The extension joins the metasternum caudad of the coxae.

The internal surface of the mesosternum (Plate VII, fig. 7, ms. ster.)

shows a furcate process arising from the anterior portion of the concavities

occurring in the coxal cavities. It consists of two anterio-dorsal pointing

arms forming the so-called entosternum, or mesoentosternum, of the

mesothorax. These arms are supported near their middle by a chitinous

of the cephalic margin of the mesosternum.

The Metathorax

Metatergum — (Plate VIII, figs. 1 and 2). The four typical tergal re-

gions are present in the metathorax. The praescutum (prs. ph.) is repre-

sented by three distinct pieces, a large semi-oval median prephragma or

praescutal phragma separated from the scutum by a large membranous

area and two lateral parts (praes.) which support the large praescutal

phragma. The scutum (set.) is composed of two large lateral halves

separated by the notal groove (n. g.) containing the scutellum. The scutal

halves are divided diagonally into an anterior and posterior region. The

14 WM. p. HAYES

anterior region bears laterally the anterior notal wing process (a. n. p.)

and the posterior region (the so-called "scapulaire posterieure^' of Straus

Durckheim) carries laterally the posterior notal wing process (p. n p.)

and the axillary cords (ax. c). The diagonal line of demarkation causing

the division of the scutal halves is the outer evidence of an internal ridge

(Plate VIII, fig. 2, d. rd.) on the interior surface of the scutum. The meta-

scutellum (Plate VIII, fig. 1, scl.) is a two-lobed piece at the posterior

median angles of the scutum. It elongates cephalad to form a tongue-like

process, which lies in the notal groove and is limited anteriorly and dorsaUy

by the membrane separating it from the praescutal phragma and internally

by the entodorsum or V-shaped ridge (ent. d.) on the internal aspect of

the metatergum. The postscutellum (pss.) is a large, irregular piece lying

immediately behind the scutellum and scutum. It bears the post-phragma

(post, ph.), is inflexed mesad to furnish attachment for several muscles and

also bears the membrane which connects the thorax with the abdomen.

The lateral edges are inflexed caudad of the alar membranes and articulate

with the epimera of the metathorax.

The Wings. — (Plate VII, fig. 9). The second or metathoracic pair of

wings, which are membranous, are borne on the metathorax and are

inserted between the metatergum and the metapleura in the alar mem-

branes. In a state of rest the wings are transversely folded under the

elytra and in flight extend nearly at right angles to the body. The wings

are articulated to the body by four axillary sclerites (Plate VII, fig. 10,

1 ax., 2 ax., 3 ax., 4 ax.) similar to those described in Mclolontha (Straus

Durckheim p. 109), one of which (4ax) according to Snodgrass (1909, p.

545) is an accessory plate not corresponding to the fourth axillary in other

forms. The first axillary lies laterad of the scutum and its anterior outer

margin abuts the basal enlargement of the subcostal vein of the wing.

Between the first axillary and the bases of the radius and medius lies the

second axillary which is partly overlapped by the first. The third is larger

than the second, and lies at the bases of the cubital and anal veins, while

the fourth axillary is quite small and lies l)clwcen the first and third axil-

laries.

The McUiplciira.--{V\-AW VHl, figs. ?> and 4). The metapleuron is

composed of two principal sclerites homologus to those of the mesopleuron

— the metacpisternum and metaepimeron — each of which is subdivided

into two regions. The lower division of the e])isternum or katepistcrnum

(keps.) is an irregular semioval j^iece attached to the lateral margin of the

metasternum. Its dorsal and posterior margins are connected to the

lower edge of the epimeron. Dorso-anteriorly the episternum exhibits the

second subdivision or anepisternum (aeps.). This is an irregular sha]ied

piece bordering on the cephalic edge of the epimeron and to which is fused

the lower jmrt of the preparapterum (pptm.) which is likewise fused with

EXTERNAL MORPHOLOGY OF LACHNOSTERNA CRASSISSIMA 15

the base of the wing process (w. p.)- Internally the preparapterum bears

a large muscle disc or pronator disc (pn. d.).

The epimeron is divided into two parts, the katepimeron and the anepim-

eron. The latter (aepm.) lies immediately above the katepisternum and

is elongated anteriorly to form the wing process (w. p.). The katepimeron

(kepm.) is a quadrilateral piece lying caudad of the anepimeron. Just

above the epimeron is the alar membrane in which is located the wing

axillaries. The epimeron is connected posteriorly by an articulation with

the postscutellum on its lateral edge. On its inner surface the suture

between the episternum and epimeron is extended to form the pleural

ridge which elongates into an adf ureal process (pi. a.) that rests on the

lateral arm of the mesoentosternal furca. The ventral end of the pleural

ridge extends to the coxa.

The Metasternum. — (Plate VII, figs. 6 and 7). The metasternum

occupies the lower surface of the metathorax. It is much the same in

shape as that of the mesothorax, but considerably larger and lies between

the meso- and metacoxse. On the internal surface of the sternum (Plate

VII, fig. 7) is the large endosternum projecting dorsally (Plate VII, fig. 8).

It consists of two laterally projecting arms which furnish support for the

adfurcal processes of the entopleura and a large and somewhat pointed

cephalad projecting arm. A manifestation of this structure is discernible

on the outer surface of the metasternum in the form of a faint mid-ventral

line.

The Metathoracic Legs

The metathoracic legs are different in structure, especially in the form

of the tibiae from the prothoracic legs which, as mentioned before, are

remarkably adapted for burrowing. While this modification is not present

in the hind tibiae, broadly speaking the metathoracic legs are quite similar

to those of the mesothorax which for this reason are not treated in this

discussion. In the metathoracic legs a well marked sexual difference, to

which reference has been made, is apparent in the distinctly longer tarsi

of the male. The trochantin of the metathoracic legs is absent, although

it is to be found both in the pro- and mesothoracic pairs.

The Coxa. — (Plate VIII, fig. 5, ex.). The coxa of the metathoracic

leg is attached to the posterior margin of the ventral surface of the meta-

thoracic segment, and likewise serves as a place of attachment for the

intersegmental membrane lying between the thorax and abdomen. Ex-

ternally it presents a flattened surface in the same plane as the metasternum

and like the coxa of the prothoracic legs is more or less immobile, except in

a semi-rotary manner. It lies at right angles to the longitudinal axis of

the body and extends from the elytra at the lateral margin to the mid-

ventral line. Internally it presents a hollow arrangement near the opening

16 WM. p. HAYES

of which is a chitinous ridge or infolding that permits the attachment of the

flexor muscles.

The Trochanter. — (Plate VIII, fig. 5, tr.). The trochanter of the meta-

thoracic leg is similar to that of the two other pairs of legs. It lies between

the coxa and femur and is triangular in shape.

The Femur. — (Plate VIII, fig. 5, f.). The femur is slightly longer than

the coxa. It is somewhat flattened with rounded edges, tapering toward

the distal end where it articulates with the tibia.

The Tibia. — (Plate VIII, fig. 5, t.). The tibia of each meso- and meta-

thoracic leg differs from that of the prothoracic leg in that there is no

flattened modification for digging and burrowing as is present in the front

leg. The proximal end articulates with the femur and the distal end with

the tarsus, where it is slightly broadened and bears two sharp spurs vary-

ing in size in the two sexes. These have been described in the paragraph

on Sexual Dimorphism.

The Tarsus. — (Plate VIII, fig. 5, tar.). The tarsus is similar to that

of the other legs in having five segments. The terminal one has two sharp

claws, each bearing a median tooth.

The Abdomen

The abdomen of Lachnosterna almost equals in volume the remaining

portions of the body, and is directed in the horizontal plane. At its base

it equals the thorax in size, to which it is attached throughout its complete

circumference. Dorsally, it is connected by a membrane to the postscutel-

lum and thereby conceals the postphragma. Ventrally, it is joined to the

posterior edge of the internal opening in the metathoracic coxae.

Concerning the number of evident (not actual) abdominal segments in

this species, there are six ventrally and eight dorsally, while the actual

number is perhaps eight ventrally and nine dorsally (Plate VIII, fig. 8).

The ninth or terminal segment is reduced in size with only the ventral

portion apparent externally, while the dorsal part is modified to form an

infolding within the anal opening and is not visible from the exterior. Each

segment, with the exception of the first, consists of two principal parts, a

dorsal or tergal region, and a ventral or sternal area. The dorsal and

ventral sclerites are united laterally by a membrane which permits dilation

and contraction of the abdomen. Posteriorly, the membrane disappears,

leaving no such separation between the terga and sterna of the terminal

segments.

The Terga. — Nine terga are present. The elytra in the state of rest

cover the first six terga which are only slightly chitinized. There is

evidently not the necessity of heavier chilinizalion which characterizes

the remaining or unprotected parts of the body. These terga are united

by comparatively wide membranous areas which are larger laterally than

EXTERNAL MORPHOLOGY OF LACHNOSTERNA CRASSISSIMA 17

near the median region. The two remaining visible terga (seventh and

eighth) are not protected by the elytra and are consequently densely chitin-

ized as is also the ninth which has disappeared within the anal opening.

The seventh and eighth are the widest tergites. They make up the pygi-

dium and are more closely fused than the anterior terga.

The Sterna. — The first sternite has disappeared and only a rudiment of

the second is present which is covered by the metathoracic coxae. The

remaining segments are more heavily chitinized than the second, are closely

fused to each other, and permit of no movement as in the terga. The male

sternites (Plate VIII, fig. 6) are somewhat flattened and the eighth sternite

has a rounded fovea which is absent in the female (Plate VIII, fig. 7).

These differences, due to sex, have been mentioned in the discussion of

sexual dimorphism.

The Spiracles. — There are nine pairs of spiracles, two thoracic and

seven abdominal. The first pair of prothoracic spiracles is located ven-

trally, and each spiracle is suspended in the membrane which unites the pro-

thorax to the mesothorax. The second pair is not visible externally but is

to be found in the suture between the mesoepimeron and the metaepister-

num. The third pair, or first abdominal pair, lies dorsally in the mem-

brane between the metathorax and the first abdominal segment. The next

five pairs are found on the ridge formed between the tergites and ster-

nites. These six pairs of abdominal spiracles are covered by the elytra.

The last, or posterior pair, is exposed below the elytra on the seventh

abdominal segment and lies in the suture between the sternum and tergum.

The Genitalia. — The genitalia properly speaking are perhaps more con-

cerned with internal anatomy and should be discussed in such a treatise.

However, they possess chitinous structures which are tegumentary in

nature and will, therefore, be briefly discussed here, chiefly because of their

importance as specific characters in taxonomy.

In the male (Plate IX, figs. 2 and 4) a heavily chitinized semicylindrical

sheath or box, termed the telum (Plate IX, fig. 4, te.), surrounds the true

membranous penis. The posterior end is shown in perspective in the

drawings showing its relation to the claspers which surmount the telum and

in this species are rather symmetrical. These claspers (Plate IX, fig. 2,

els.) are of much taxonomic importance.

Underlying the telum is a small Y-shaped chitinous structure (Plate

IX, fig. 3). Posteriorly, the branching arms are bent dorsally and to them

is attached the membrane which constitutes the anterior region of the

cloaca. The membrane is also attached to the inner margin of the last

ventral and abdominal segments. Anteriorly, this structure extends into

the body as far as the sixth ventral abdominal segment.

The female genitalia (Plate IX, fig. 1) are shown in three views. The

organ consists of a pair of broad inferior plates (inf. pi.) which surround a

18 WM. p. HAYES

smaller pair of superior plates (sup. pi.) somewhat cylindrical in shape and

strongly divergent.

Literature Consulted

1908 Breed, R. S.. and Ball, E. F.

The interlocking mechanisms which are found in connection with the elytra of

Coleoptera.

Biol. Bull., 15:289-303. Abstract, Proc. of Seventh Int. Zool. Cong., Cambridge,

Mass. 1912.

1902 CoMSTOCK, J. n., and Kochi, C.

The skeleton of the head of insects.

Amer. Nat. 36:13-45, 29 figs.

1909 Crampton, G. C.

A contribution to the comparative morphology of the thoracic sclerites of insects.

Pros. Acad. Nat. Sci., Phiia., 61:3-54, 4 pis.

1907 Hardenberg, C. B.

A comparative study of the trophi of Scarabaeidae.

Trans. Wis. Acad. Arts and Sci., 15:548-602, 4 pis.

1902 Kadic, O.

Studien iiber das Labium der Coleoptern.

Jena Zeitschrift fiir Nat. Wissenschaft, 36:207-228.

1901 KOLBE, H. J.

Vergleichendmorphologische Untersuchungen an Coleoptern nebst Grundlagen zu

einen System and zur Systematik derselben.

Arch. Naturg. 67:89-150, 2 pis.

1892 Smith, J. B.

The mouth parts of Coprls Carolina; with notes on the homologies of the mandibles.

Trans. Amer. Ent. Soc. 19:83-87, 2 pis.

1906

Explanation of terms used in entomology.

Brooklyn Ent. Soc. Separate, p. 154 and plate IV'.

1906 Snodgrass, R. E.

A comparative study of the thora.x in Orthoptera, Euplexoptera and Coleoptera.

Proc. Ent. Soc. Wash., 9:95-108, 4 pis.

1909

The thorax of insects and the articulation of the wings.

Proc. U. S. Nat. Mus., 36:511-595, 6 figs., 30 pis.

1909

The thoracic tergum of insects.

Ent. News, 20:97-104, 1 pi.

1828 Straus-Durckiieim, H.

Considerations generales sur I'anatomic comparec dcs aiiimaux articules.

Paris, pp. 19 434, 10 pis.

List of .Xbbkkviations

EXTERNAL MORPHOLOGY OF LACHNOSTERXA CRASSISSIMA

19

20

WM. P. HAYES

fac

Pi.ATi: I

Fig. 1. I'ront view of head.

Fig. 2. Lateral view of head.

Fig. 3. Ventral view of head.

F"ig. 4. .Antenna of male.

Fig. 5. Antenna of female.

Fig. 6. F'acets of the compound eye.

EXTERNAL MORPHOLOGY ^ LACHNOSTERNA CRASSISSIMA 21

DaJt.pC

Plate II

Fig. 1. Epipharynx and internal aspect of clypeus and labrum.

Fig. 2. Gula and Labium.

Fig. 3. Left maxilla.

22

WM. P. HAVES

Plate III

Fig. 1. Side view of right mandible.

Fig. 2. Apodeme of mandible.

Fig. 3. Membrane and hairs at base of mandible.

Fig. 4. Portion of cross section thru molar.

Fig. 5. Hairs and setx from upper membrane of mandible.

Fig. 6. Inner surface of mandible.

EXTERNAL MORPHOLOGY OF LACHNOSTERXA CRASSISSIMA

23

^ _ CS^"-'

ful .hyp

nyp

fMl.hyp

Plate IV

Fig. 1. Labium from within showing hypopharynx and fulcrum h)T3ophar>'ngeum.

Fig. 2. Junction of arms of fulcrum hypopharyngeum, ventral aspect.

Fig. 3. Lateral view showing relation of fulcrum hypopharyngeum to the pharynx.

24

\VM. P. HAYES

oo.for

tenti

post .a

Ua

ant. a

3

Plate V

Fig. 1. Tentorium — looking througli occipital foramen.

Fig. 2. Anterior view of tentorium.

Fig. 3. Dorsal view of tentorium.

F'ig. 4. Left side of cervicum (dorsal margin to the right).

EXTERNAL MORPHOLOGY OF LACHNOSTERNA CRASSISSIMA 25

pro.ster

post.ph

e 8 . aph

cx.ph

t.cl

Plate VI

Fig. 1. Dorsal view of prothorax.

Fig. 2. Ventral view of prothorax — coxae removed.

Fig. 3. Anterior aspect of prothorax.

Fig. 4. Left elytron.

Fig. 5. Articulation of elytron.

Fig. 6. Right prothoracic leg.

26

WM. P. HAVES

praes.ph

pfaea.ph

10

Plate VII

Fig. 1. Dorsal view of mesothorax with portion of right elytron.

Fig. 2. Left lateral aspect of mesothoracic tergum.

Fig. 3. Internal aspect of mesothoracic tergum.

Fig. 4. Left mesopleuron, external aspect, coxa ex situ to show trochantin.

Fig. 5. Left mesopleuron, internal aspect.

Fig. 6. Meso — and metastcrna, external aspect.

Fig. 7. Meso — and metastcrna, internal aspect.

Fig. 8. Metathoracic endosternum.

Fig. 9. Metathoracic wing.

Fig. 10. Axillary sclerites of the metathoracic wing.

EXTERNAL MORPHOLOGY OF LACHNOSTERNA CRASSISSIMA

pi- s . ph

P^s.ph

27

ppin

Plate VIII

Fig. 1. Metatergum, external aspect.

Fig. 2. Metatergum, internal aspect.

Fig. 3. Left metapleuron, external aspect.

Fig. 4. Left metapleuron, internal aspect.

Fig. 5. Right metathoracic leg.

Fig. 6. Ventral aspect of abdomen, male.

Fig. 7. Ventral aspect of abdomen, female.

Fig. 8. Left lateral aspect of abdomen, male.

28

WM. P. HAYES

^J— sup.pl —

Fig. 1.

Fig. 2.

Fig. 3.

Fig. 4.

Fig. 5.

Plate IX

Female genitalia, dorsal, ventral and lateral aspects.

Male genitalia, arranged perspectively.

Y-shaped supporting structure of cloaca and telum.

Male genital organ with telum in place.

Dorsal aspect of Lachnosicrna crassissima.

THE RESPIRATORY MECHANISM IN CERTAIN AQUATIC

LEPIDOPTERAi

By

Paul S. Welch

Introduction

According to one of the current theories, insects arose from a terres-

trial ancestry and the aquatic habit, wherever manifested, was secondarily

acquired. The general appUcation of this theory to all aquatic insects is

sometimes questioned but there seems to be an almost universal agreement

that many of the higher orders, including the Lepidoptera, are preeminently

terrestrial in organization and that their aquatic representatives display

an evolution superimposed upon a terrestrial background. These insects,

invading water, had certain vital problems to solve, respiration being

among the first, and the diverse but successful adaptations offer interesting

material for study.

The writer has found the aquatic Lepidoptera favorable for the study

of larval adaptations for the following reasons:

1. While the Lepidoptera constitute a large, well-defined order, the

insignificant number of aquatic forms made it possible to examine most

of the American species.

2. Although inconsequential numerically, they manifest adaptations

to aquatic Kfe as perfect and as diverse as many of the more conspicuous

groups.

3. Heterogeneity in the methods of solving aquatic problems may ap-

pear even among species of the same genus.

4. The abundance of some species in certain environments has provided

ample material for extensive observations and experiments.

5. This group of aquatic insects is practically unstudied from the

morphological, physiological and ecological aspects.

Excluding the semi-aquatic forms, aquatic Lepidoptera fall into two

general classes when considered from the point of view of larval respira-

tion: (1) Those which are devoid of any special respiratory organs, and

secure the requisite oxygen from the atmosphere by some physiological

adaptation, from the dissolved supply in the water, or through a combina-

tion of both; and (2) those with special morphological devices in the form

of gills. In the first class belong such forms as Bellura melanopyga, Nyniph-

' Contribution from the University of Michigan Biological Station, and the Zoological

Laboratory, University of Michigan.

29

30 PAUL S. WELCH

ula icciusalis, and Pyrausta pemtalis, while Nymphula maculalis, N,

obscuralis, and Catadysta fulicalis are typical representatives of the

second.

Acknowledgments

The writer wishes to express his indebtedness to Professor S. A. Forbes,

Professor J. G. Needham and Mr. J. T. Lloyd for certain materials speci-

fied on later pages. Use has been made of data accumulated by two of the

writer's graduate students, Miss Ethelynn Hopkins and Mr. Jennings

Hickman. During the summer of 1919, Miss Hopkins studied briefly

the tracheation of certain instars in Nymphula maculalis and Nymphula

icciusalis, using living material. Later, Mr. Hickman examined the gills

of Nymphula maculalis, using serial sections of preserved specimens. The

work of both of these students has been repeated and extended by the

writer.

Material

The principal objects of the work on which this paper is based were

to determine the structure and mode of function of the respiratory organs

developed in this group; to study the degree of change from the original

terrestrial organization of the larvae; to get some measure on the efficiency

of 'Special organs; to examine the various instars with respect to the respira-

tory problem; to test certain doubtful statements in the literature; and if

possible to determine something as to the course of evolution in such a

hmited group.

In order to accomplish these ends it was necessary to study as many

representatives of the two above mentioned classes of aquatic Lepidoptera

as possible. Owing to the abundance of several species in the vicinity of

the University of Michigan Biological Station, repeated observations and

experiments have been made during the past four summers. Some of the

necessary preliminary work on habits and life-history has been published

already (Welch, 1914, 1915, 1916, 1919). The material used most exten-

sively in the present work is as follows:

Non-gilled Larvae

Nymphula icciusalis Wlk. — This species occurs abundantly at Douglas

Lake, Michigan, and several instars have been studied in both living and

preserved form.

Nymphula oblitcralis Wlk. — Preserved material of the larvae of this

species was secured from the collections of the Illinois Natural History

Survey through the courtesy of Professor S. A. Forbes. The life-history

of this species was described by Hart (1895, pp. 176-180) and the speci-

mens used were from Hart's collections and identified by him. Living

material has not been available.

RESPIRATORY MECI^NISM IN LEPIDOPTERA 31

y ymphula sp. — Larvae of a species of XympJiula occasionally appeared

at Douglas Lake on the leaves of the yellow water-lily but it has not been

bred to the adult stage and its specific identity is not known.

Gilled Larvae

N ymphula mactilalis Clem. — Living material in all stages of the life-

history including larval instars is available in abundance at Douglas Lake,

Michigan, during July and August. Observations were also made on living

specimens at Lake Oneida, N. Y., during the summer of 1916. Liberal use

was made of preserved material in connection with the morphological work

on the spiracles and gills. The biology of this species (Welch, 1916) has

already been described.

Xympliula obscuralis Grt. — Preserved larvae were received from the

Illinois Natural History Survey. Hart (1985, pp. 167-173) reported on the

life-history of this form and the specimens sent to the writer were from

Hart's collections and identified by him. No living material was available.

Cataclysta fulicalis Clem. — Preserved larvae were sent to the author

by Professor J. G. Needham and Mr. J. T. Lloyd, from collections made in

the vicinity of Ithaca, New York. These gilled caterpillars have been

described by Lloyd (1914, 1919) and are conspicuously different from the

non-gilled Cataclysta forms in other parts of the world. Living material

was not available.

The Tracheal System

Since the gilled caterpillars represent the greatest progress in the

direction of special aquatic respiratory adaptation and since the gap

between the gilled and non-gilled forms is without intergrades, it was of

interest to compare the tracheal systems of species representing these two

classes and to seek light on the following questions: (1) Do the non-gilled

forms possess a system of tracheation built on the plan of terrestrial lepi-

dopterous larvae? (2) Do the gilled forms have the same system of tra-

cheation found in the non-gilled aquatic forms? or (3) Has the acquisition

of gills been accompanied by noteworthy changes in the fundamental

tracheation?

These questions are significant in connection with certain facts of gen-

eral habits and life-history. There is little or no doubt at present that the

non-gilled forms once included in the genus Hydrocampa and the gilled

forms once included in the genus Paraponyx really comprise one genus,

N ymphula, and they are usually so treated. In Douglas Lake and other

similar lakes studied by the writer, it is commonplace to find the gilled

caterpillars and non-gilled caterpillars thriving side by side in the same

vegetation beds, subject to identical environmental factors. In N . macula-

lis the first instar is devoid of gills, these appearing only in the second

instar, when forty gill-filaments come into existence. The gill-filaments

32 PAUL S. WELCH

increase in number with each succeeding molt until the final larval instar

has an equipment of over four hundred.

Non-gilled Larvae

A study was made, using living material, of the arrangement and dis-

tribution of the tracheae in the different larval instars of .Y. icciusalis,

giving special attention to the early instars and the full-grown larva.

Taking into account only such tracheae as appear under the higher powers

of the binocular microscope and the medium powers of the compound

microscope, and using only fresh preparations from which none of the

air had been lost, it was possible to diagram the fundamental structure of

the tracheal system and to make comparisons not only in the different

instars but also with other species.

It has thus been found that the ground plan of arrangement and dis-

tribution of tracheae is essentially that which characterizes the terrestrial

larvae. Deviations and minor variations appear but none of them seem

to bear any significant relation to the acquired habits of the larvae. No

important changes of any kind appear as later instars are reached.

Gilled Larvae

Detailed studies of tracheation in the larval instars have also been

carried on with ^Y. maculalis using fresh, living material. In the first

instar the system is almost a dupUcate of that in the first instar of N . icciu-

salis. In the second, the appearance of gills is accompanied by no conse-

quential change, the gills being supplied by short direct branches from the

main longitudinal tracheae. Also in the rapidly increasing gill complexity

of the later instars there is no deviation which the writer can recognize as

having any significance in relation to the aquatic habit or to the acquisition

of gills.

The results of this part of the work seem to show that fundamentally

these aquatic caterpillars have retained the original terrestrial form of body

tracheation and that the gill tracheation is a system superimposed upon

the one already present in the whole group. Gills have therefore been

developed with a minimal change of the original tracheation. It would

appear, if these conclusions are well taken, that the larval type represented

by that of N . icciusalis is the older one phylogenetically and that the gilled

caterpillar has a more recent origin.

Structure of the Gills

A detailed study of the structure of the gills in all of the species avail-

able was made by means of longitudinal and transverse sections of pre-

served material and by the examination of living material, whenever the

latter could be secured. The relative transparency of the living material,

especially when submerged in dilute glycerine, or some of the oils used for

RESPIRATORY MECHANISM IN LEPIDOPTERA

33

clearing made it possible to study certain features more readily than in

sections, particularly the distribution of tracheoles. Sections cut 6 microns

thick and double stained in haemotoxylin and eosin gave satisfactory re-

sults. High magnification was often required for the examination of sec-

tions, particularly for the study of the histological features, and for the

more critical and difficult features a 1 .9 mm. oil immersion fluorite objec-

tive was used in connection with a monobinocular microscope.

The Gill-wall

A study of the gill-wall shows that it is essentially a continuation of

the body- wall, having the same set of layers. In order to determine whether

any differences in thickness appear, a number of measurements were made

of the various layers of the gill and of adjacent portions of the body- wall,

the averages being given in the following table. All of these measurements

were made on specimens in the last larval instar. Measurements in this

and succeeding tables are expressed in fractions of a miUimeter. The

terms epidermis and dermis are used instead of the primary cuticula and

secondary cuticula of some authors.

It thus appears that there is a distinct reduction of thickness in the

walls of the gills as compared with the body- wall, this reduction occurring

mainly in the dermis. The basement membrane is so delicate that it has

been left out of account in all measurements of body-wall and gill-walls.

The Gill-cavity

The interior of each gill is merely a cavity (Pi. X, fig. 6) enclosed

by the walls described above and containing certain structures to be

discussed in another connection. This cavity extends continuously from

base to tip and lacks completely, in the species examined, the alveolar type

of tissue which appears within the gills of some insects. The size of this

34 PAUL S. WELCH

cavity depends entirely upon the dimensions of the gill as a whole and has a

direct connection with the haemocoele, in fact, it is a continuation of the

haemocoele. In the Nymphula group, the gills are relatively large, have a

spacious gill-cavity, and the passage from the haemocoele to the gill-cavity

is broad, while in Catadysta fnlicalis the gill as a whole is small and slender,

the wall thick, the gill-cavity much reduced, and the passage from the

haemocoele to the gill-cavity often smaller in diameter than that of the

gill-cavity. For example, in one series of eighteen measurements the aver-

age diameter of the gill-cavity was 0.0054 mm. while the average diameter

of the opening into the haemocoele was 0.0039 mm.

Contents of the Gill-cavity

Tracheae

Nymphula maciilalis. — As previously mentioned, the gills in this species

are supplied with tracheal branches arising directly from the adjacent

main longitudinal tracheae. Since the gills become branched in later instars,

the supplying tracheae branch correspondingly. Each filament, therefore,

has one main trachea, axial in position, which extends from the base almost

to the tip, gradually decreasing in diameter distad. A very few instances

of two tracheal branches entering a gill-filament were observed, both

extending well towards the tip of the filament and both giving rise to

tracheoles.

The origin and distribution of the tracheoles were best studied in hving

material, although certain data were confirmed by means of serial sections.

At frequent but irregular intervals (PL XI, figs. 9, 10, 11) along the sup-

plying trachea short tracheoles arise singly, extending ectad to the inner

surface of the gill-wall and giving off numerous fine branches, all of which

break up into very minute tracheoles and have a rather definite arrange-

ment as follows. These tiny tubes all extend longitudinally, proximad and

distad, very near or in contact with the ental surface of the gill-wall, and

approximately parallel to each other, so that the periphery of the gill-

cavity is bounded by a thin zone composed of countless, minute, parallel

tracheoles. The terminal tracheoles of each individual tuft intermingle

with those of the adjacent tufts but also in an approximately parallel

fashion. All attempts to determine the character of the terminations of

these tracheole endings, using the best preparations and the highest mag-

nifications, hav/' thus far been futile. In living material and in whole

mounts they appear to unite with the basement membrane and serial

sections confirm this conclusion, but nothing further can be said as to the

exact relation to the gill-wall. No tracheoles were found lying free in the

gill-cavity. The profusion of these tracheoles, intimately related to the

entire inner surface of the gill-wall, points definitely to the principal func-

tion of these body projections.

RESPIRATORY MECHANISM IN LEPIDOPTERA 35

Nymphula obscuralis. — Since it has not been possible to study living

material of this species, the tracheation of the gills cannot be so definitely

described. However, serial sections show a type of structure closely resem-

bling that of N . maculalis. It is probable that both species have systems

which are very similar.

Catadysta fulicalis. — Serial sections including all parts of the body of

the larva show no tracheation (PL X, fig. 5) of the gills. No branches of

the body tracheal system approach the bases of these organs and in no

sense are they to be regarded as tracheal gills.

Body Fluids

In living specimens of N . maculalis, it is easy to observe the movement

of fluid, not only within but also into and out of the gill-cavity, thus giving

added proof of the continuity of this cavity with the haemocoele. Blood

corpuscles can be detected in this fluid. Sections confirm the observations

on living specimens, showing that the gill-cavities invariably contain

haemocoele fluids. Preserved material of .V. obscuralis yielded similar

results.

The small attenuated gill-cavities of Catadysta fulicalis contain only

the remains of fluid originating from the haemocoele.

Discussion

It thus appears, from the point of view of structure alone, that two

distinct gill types occur in aquatic larvae of Lepidoptera: (1) combination

tracheal-blood gills, and (2) blood gills.

As already pointed out, the profusion of tracheoles in each gill-filament

in N. maculalis indicates the respiratory nature of these organs. With

its equipment of over four hundred gill-filaments the mature larva appar-

ently has more than ample provision for respiration, especially since these

larvae live in surface water rich in dissolved oxygen, and often in vegeta-

tion beds which contribute additional oxygen. This gill equipment is also

striking in view of the fact that certain non-gilled Nymphula larvae thrive

in identically the same external conditions. Nymphula obscuralis, accord-

ing to Hart (1895, p. 170), has an average of four hundred and eighty-four

gill-filaments. In Catadysta fulicalis, the number is smaller, the full-

grown larva having about one hundred and twenty unbranched gills.

Both blood gills and tracheal-blood gills are known to occur in limited

numbers in other orders of insects. There is no special difficulty in under-

standing the mode of functioning of the ordinary tracheal gill, but in the

combination described above and in the blood gill of the Catadysta type,

certain problems arise, first of which is the nature of the relation, if any,

of the blood (the body fluid which circulates in the gill-cavities) to the

transportation of oxygen. In the blood gills of certain chironomids, the

36 PAUL S. WELCH

blood contains haemoglobin and with this carrier present the gills have

definite significance. In the larvae of N. maculalis and in certain other

insects having similar gills, some carrier other than haemoglobin seems

necessary to enable these structures to function as gills. The actual dis-

covery of invisible carriers has not yet occurred. Muttkowski (1920,

1921a, 1921b, 1921c) suggests that possibly haemocyanin may constitute

such a carrier. Rose and Bodansky (1920) demonstrated the presence of

copper in a number of marine organismiS and Muttkowski (1921a) found it

in a large number of animals representing six phyla. The last named in-

vestigator holds that "Copper is found in insect blood in quantities com-

parable to that of crayfish blood. Its role is therefore interpreted as being

identical, — namely that it serves as the nucleus of a respiratory protein,—

hemocyanin. Insects, therefore, have two sources of oxygen,— atmospheric

air led directly to the tissues by way of the tracheae, and fixed oxygen ar-

ried by the respiratory protein of the blood." Possibly this is a hint in the

right direction and invisible oxygen carriers in insect blood may soon be

identified.

Since the circumstances seem to demand the presence of some oxygen

carrier, the question arises concerning the mode of functioning of the com-

bination gill. Does such a gill have two separate and distinct methods of

securing and distributing oxygen? The position and distribution of the

tracheoles are such that there seems to be no ground for assuming any rela-

tion to the blood as an intermediary between them and incoming oxygen.

Perhaps the tracheal system might function as completely if the gill-cavity

were filled with alveolar tissue instead of blood. On structural grounds

alone, it appears possible that two distinct methods could exist side by

side. It might be suggested that in N. maculalis and N. obscuralis the

gills are really tracheal gills and that the presence of blood in the gill

cavity is entirely incidental, but such a suggestion loses weight when C.

fulicalis is considered since its gills, if they function at all, must do so

through the intermediation of the blood. It has not been proven absolutely

that these lateral outgrowths in C. fulicalis arc functional gills and as

respiratory organs they might be questioned completely. Such a view

would render similar organs in other orders of insects devoid of respiratory

significance and, pending further investigation, it would seem that circum-

stantial evidence points rather definitely to the conclusion that these organs

do function in res])irati()n

Judging entirely from the structure of these gills, a contrast appears

between the Nymphula group and Cataclysta which may indicate difference

in degree of efficiency. In the former, the larger number of gills, the rather

spacious gill-cavities, the thin gill-walls, and the profuse tracheation all

suggest an efficient equipment. In the latter, however, with only about

one hundred and twenty gills, with the very small gill-cavities connected

RESPIRATORY MECHANISM IN LEPIDOPTERA 37

with the haemocoele by still smaller lumina, with no traces of tracheation,

and with the conspicuously thick gill- walls, the effectiveness of the system

seems much smaller.

The presence and absence of gills within the genera Xymphula and

Cataclysla and the existence of distinctly different gill types in these two

closely related genera give added support to the theory of the independent

origin of the various aquatic insects, emphasizing the fact that in these

animals types of adaptation and genetic relationship may have no close

correlation.

The Spiracles and Connecting Tr.aciieae

The secondarily acquired nature of aquatic habits and structures

naturally directs attention to the character of the spiracles. In the gilled

forms, is the gill system superimposed upon an unmodified holopneustic

tracheation, or have modifications occurred leading towards suppression

of the spiracular equipment? In the non-gilled forms which lead a sub-

merged existence, has the characteristically terrestrial holopneustic trachea-

tion been modified? A common statement appears in the literature to the

effect that many nymphs and larvae living in water have apneustic trachea-

tion, breathing directly through the skin or by means of gills. It is also

pointed out that between the completely apneustic and the typical holop-

neustic tracheation a variety of intermediate stages exists. The gilled

larvae of certain Xymphula species have been described as having apneustic

tracheation in which the spiracles are closed, and the spiracular branches

(stigmatal branches) have become solid cords. The writer has searched

in vain for any thoroughgoing morphological work bearing on this subject.

Among workers who have studied Old World species of Nymphula three

of the most recent might be mentioned. Rebel (1899) made some studies

on Nymphula iParaponyx) stratiotata and states that the tracheation is

apneustic. Portier (1911) in a voluminous paper dealing with several

aquatic insects, carried on some physiological experiments with larvae

of XympJmla stratiotata and claims to have shown that (a) larvae sub-

merged for five minutes in olive oil colored with alcanine showed no oil in

the tracheae; (b) that under the binocular microscope the spiracular

trunks did not have the aspect characteristic of air filled tracheae but

looked like heavy cords; (c) that larvae suffered no effects from submer-

gence in oil, but a small geometrid larva so treated became inert and oil

was found in its tracheae; (d) that larvae were perfectly normal after

fifteen minutes submergence in soapy water, but the geometrid larva so

treated became apparently dead in three minutes; (e) that larvae immersed

in olive oil, ether, and alcanine became aenethetized after one minute, but

microscopic examination showed no penetration of the colored oil, recovery

occurring when returned to water; (f) that larva treated as in (e) for

3S PAUL S. WELCH

twenty hours did not show the tracheal system invaded; (g) that a larva

placed under reduced air pressure showed bubbles of gas gradually form

on the surface of the body where the integument is thinnest but none

formed about any of the spiracles. From these experiments and without

morphological confirmation Portier concluded that the spiracles of the

larva of N. stratiotata are closed and functionless. Wesenberg-Lund (1913,

p. 126) states that in N . ("Paraponyx'^) stratiotata the spiracles are func-

tionless but no evidence is given in support of this conclusion.

Observations and Experiments of Nympiiula Maculalis

Attention was first directed to the spiracles by evidence that these

larvae can exist out of water for considerable periods of time. While it is

common for the pupa to be formed on the lower, submerged surface of a

water-lily leaf, the full-grown larva sometimes crawls out of the water,

onto the upper leaf-surface and there forms the pupa. In order to con-

struct the silken covering, tie down the case to the leaf, and transform into

a pupa, a considerable period of time must be spent out of water. Mature

larvae placed in containers without food and with little or no moisture often

lived from four to eight days. Larvae, in containers with food material

and just enough water to keep the surrounding air moist, were allowed to

gradually dry up. In such cases the gills became dried and black, but the

larvae lived about fourteen days. It is thus evident that these caterpillars

can exist for days apart from water and respire by means other than gills.

Furthermore, it seems unlikely that this can be accounted for on the basis

of cutaneous respiration. These results led to a critical morphological

examination of the spiracles and their connecting tracheae in A', maculalis,

a study which was later extended to include all of the strictly aquatic

caterpillars available for examination.

Morphology of the Spiracles and Connecting Tracheae

A detailed morphological study of the spiracles and their connecting

tracheae was made on serial sections, cut six to seven microns thick, and

double stained. Both transverse and longitudinal sections were used

and all critical points determined with a modern monobinocular micro-

scope equipped with a 1.9 mm. fluorite oil immersion objective. Since

the spiracular aperture may be narrower in one dimension than the other,

the long dimension being transverse to the long axis of the body, measure-

ments on transverse sections might lead to error if not checked on longi-

tudinal sections. In all such cases the measurements were made from the

edges of the opening, not including accessory structures, and in such a way

as to give an average of the two principal diameters of the aperture. On

the other hand, the connecting tracheae arc practically cylindrical, thus

making it possible to record measurements from any section passing

RESPIRATORY MECHANISM IN LEPIDOPTERA

39

through the center of the lumina. Mature larvae were usually used, al-

though sections of earlier instars were examined from time to time.

In this connection it should be pointed out that, as usual in lepidopter-

ous larvae, the spiracles on the meso- and metathorax are absent in all of

the species examined. In the following tables percentage of decrease,

wherever expressed, is calculated by using as the standard of comparison

the dimensions of the largest spiracle and connecting trachea (usually

those of the second abdominal segment) of the series. It must also be

understood that this is merely a convenient way of comparing the degree

of reduction of the other spiracles and tracheae and is not intended to imply

that even the largest may not have undergone some reduction themselves.

Since there is no way of determining the diminution of the largest spiracle,

if it has been reduced, it would not be possible to express the true amount

of reduction of the other spiracles on the same individual.

Nymphula maculalis

Examined externally, under magnification, nine pairs of spiracles are

observable on segments 1, and 4-11. All are minute and inconspicuous

except those on 5, 6, and 7 which are distinctly larger. The following

table comprises one set of diameter measurements which is representative

of all others made in this work:

The Larger Spiracles and Connecting Tracheae

In the larger spiracles the outer margin of the aperture bears a com-

plete set of elongated, closely set, chitinous spines (PI. X, fig. 1) which

have a rich yellow color when viewed under magnification. These spines

form an almost continuous marginal guarding device, the free ends con-

verging so that the form of the whole is that of a truncated cone. The

free ends of the spines mark the periphery of an aperture which is much

40 PAUL S. WELCH

smaller than the spiracle itself. The external cuticula extends into the

lumen of the spiracular trachea, Uning it for the entire length. However,

certain modifications appear chief among which are the distinct reduction

in thickness and the numerous filiform chitinous projections which extend

into the lumen. A continuation of the hypodermis of the body-wall con-

stitutes the major part of the wall of the spiracular trachea and shows

no significant changes in structure.

The short connecting trunk is terminated at its ental end by a well-

developed closing apparatus (PI. X, fig. 4), composed essentially of a

closing bow, a closing band, a closing lever, and an occlusor muscle. The

closing bow is a chitinous, crescentic band (PL XI, fig. 7) lying in the lining

of the lumen and extending through one-half of the total circumference.

From one end of the closing bow a similar band continues around to a point

about opposite the middle of the closing bow where it meets the end of

the closing lever. The closing lever is located at right angles to the lumen

and projects radially for its entire length, covered by an extension of the

hypodermal wall. A short, broad occlusor muscle extends from the end

of the closing lever diagonally to the free end of the closing bow. It thus

appears that the chitinous band formed in this way is absent for about

one fourth of the circumference of the lumen, thus failing to form a com-

plete ring. Thus far it has been impossible to determine the exact relation

of the chitinous parts of the closing apparatus to each other. Studies were

made using thin serial sections in all the principal planes; also by dissecting

out a portion of the body of the larva containing a large spiracle and its

related tracheal parts, placing them on a slide in strong potassium hydrox-

ide solution and boiling by holding the slide over a small flame. By the

use of this last named method the soft parts were all removed leaving only

the chitinous portions. Difficulties in tracing out certain minute portions

of this closing apparatus have not been entirely overcome either by the

kind of preparation or by high magnification. It appears, however, that

the chitinous band, forming the closing lever and the closing bow, is one

continuous structure.

From the outermost end of the closing lever a long muscle band extends

diagonally to the body-wall. On the opposite side a similar muscle ex-

tends from a point near the origin of the occlusor muscle diagonally to the

body- wall. Beyond the closing apparatus the lumen oi)ens directly into

that of the main longitudinal tracheal trunk.

The Smaller Spiracles and Connecting Tracheae. — The spiracles on seg-

ments 1, 4, 8, 9, 10, and 11 show a marked reduction in size, in fact, they

are so small that magnification is necessary to locate them definitely.

Structurally, these spiracles and their connecting tracheae difTer markedly

from those on 5, 6, and 7. At the margin of each spiracle there appears,

instead of a thick set crown of chitinous spines, a solid, continuous, chiti-

RESPIRATORY MECHANISM IN LEPIDOPTERA

41

nous rim (PI. X, fig. 3) which projects distad from the body-surface. It

would appear that in the process of reduction the spiny crown of the origi-

nal large spiracles became fused into one continuous margin. At the base

of the tiny cup thus formed the lumen becomes reduced to an extremely

fine canal which extends without change in diameter to the closing appar-

atus located well within the haemocoele. This canal is lined throughout by

a uniform, thin extension of the external cuticula, but the hair-like projec-

tions characteristic of the large spiracular trunks are here entirely wanting.

As will appear in the table, the lumen is very minute, but by means of

thin, serial sections and high magnification it has been possible to demon-

strate that it is open throughout its course. The bulk of the wall of the

spiracular trachea is composed of an extension of the hypodermis of the

body-wall, but it also has become reduced, being about one-half the

thickness of the same layer in the larger spiracular tracheae. The connect-

ing trachea has not changed in length and terminates in a closing apparatus

similar to that described for the larger spiracular tracheae, except that it

also has become considerably reduced in size. All of the parts are repre-

sented, however, and the whole closing contrivance has every appearance

of being completely functional.

It also appears that reduction in the small spiracles is not uniform, but

gradually increases posteriorad. This, however, does not seem to hold for

the connecting tracheae.

Xy mph ula obscuralis

An examination of Xymp/iula obscuralis showed a condition very simi-

lar to that in X. maculalis. While the following diameter measurements

were taken from a single mature larva, they are representative of those

for other larvae. The structural features of the spiracles, the connecting

42

PAUL S. WELCH

tracheae, and the closing apparatus, are also so similar that no description

is necessary here.

Cataclysta fulicalis

In Cataclysta fulicalis the spiracles and the connecting tracheae differ

from those of the gilled Nymphula caterpillars in that no reduction of

any kind appears, all being of practically uniform size and structure.

Some variation occurs but it is slight and apparently insignificant. The

circle of guarding spines at the outer periphery of the spiracle is less con-

vergent (PI. X, fig. 2) than in the gilled Nymphula larvae, thus forming

a wider aperture. Structurally, the spiracles, connecting tracheae, and

the closing apparatus are similar to those of the gilled Nymphula group,

slight but inconsequential deviations being present. The following table

includes a typical set of diameter measurements made on one specimen:

Segment

Spiracle .

Lumen of

trachea

11

0.031

0.038

When all measurements were averaged, the spiracles and their connect-

ing trunks were found virtually uniform in size. The larvae of Cataclysta

fulicalis have thus acquired a system of gills without accompanying

changes in the spiracular system, the latter being as completely open

morphologically as any terrestrial caterpillar.

Nymphula ohliteralis

The spiracles and connecting tracheae are distinctly open and show no

evidence of definite reduction. A certain variation appears, as will be

noted in the following table, but even the smallest found is far above

the corresponding reduced structures in N . maculalis and N. ohscuralis.

In the following representative diameter measurements taken from the

record of one mature specimen, nothing is especially noteworthy except

the large spiracle and connecting trachea on the fourth segment. Whether

the difference in size is an evidence of a slight reduction of spiracles and

connecting tracheae in the posterior segments is uncertain. The principal

features of the closing apparatus are shown in figure 8.

Segment.

Spiracle. .

Lumen of

trachea

11

0.04

0.0.^6

RESPIRATORY MECHANISM IN LEPIDOPTERA 43

Xympliula sp.

A non-gilled form, distinctly different from any other used in this

work but whose specific identity is unknown^ was examined in this connec-

tion. Sections showed all of the spiracles to be distinct, well-developed,

open, and approximately uniform in structure and size, the same being

true of the connecting tracheae. Average measurements for the whole

series are as follows, the extreme variations deviating very little from the

average: Diameter of the spiracles, 0.05569; diameter of the lumina of

the tracheae, 0.0527.

Expcrifuoits

In order to demonstrate experimentally the open condition of the

spiracles and connecting tracheae and check the morphological results,

certain experiments were made on the caterpillars N. maculalis. Larvae

dropped into hot water invariably give off one or more bubbles of gas

from the large spiracles on segments 5-7, thus showing definitely the

open condition of these spiracles and their connecting tubes. No gas was

given off from the reduced spiracles, but it does not follow that such

failure is due to complete closure since the amount of gas in the tiny

lumina is extremely slight and the expansion of the heated gas in the longi

tudinal tracheal trunks would be more likely to be released at the larger

and more open spiracles on segments 5-7.

It was found that the larvae of N. maculalis could live in commercial

kerosene for 6-7 hours. They were then submerged in kerosene colored

with Sudan III. Since the translucency of the body-wall made it possible

to trace much of tracheal system, it was easy to examine the spiracular

connections at any time and to follow the entrance of the colored liquid

into the larger spiracles. Positive evidence that these spiracles are mor-

phologically open was thus repeatedly secured. This penetration, into the

large spiracles, occurred within an hour, but entrance into the smaller

ones was much slower although ultimately the colored liquid was observed

in some of the connecting tracheae.

DlSCUSSION

It thus appears, at least in Xyviphula maculalis, X. obscuralis, and

Cataclysta fiilicalis, that in spite of the gill development, the tracheal sys-

tem of the larvae is morphologically open. The reduction of certain

spiracles and their connecting tracheae in the first two is definite and

striking but has not progressed to the place where the lumina and apertures

are completely closed. Since the previous work on this subject has been

done on unavailable foreign species it is not possible, on the basis of the

present work, to absolutely refute the statements made in the literature,

but the writer is inclined to suspect strongly that what has been found

44 PAUL S. WELCH

in N . niaculalis and .Y. obscuralis is likewise Irue of N . stratiotata and other

foreign gilled representatives of that genus. Portier's results (1911),

secured as they were without any attempt at critical morphological work,

cannot be regarded as conclusive. However, it is not inconceivable or

impossible that certain species might have progressed to the point of closing

the spiracular system but if such a condition does exist it should be demon-

strated more convincingly than has heretofore been done.

In regard to the functioning of these reduced spiracles and tracheae,

no serious question can be raised concerning the larger ones on segments

5-7 in N . maculaUs and lY. obscuralis since their size, open character of the

connecting tracheae, and structure of the closing apparatus all indicate

the possibility of normal activity. In spite of the small diameter of the

more reduced spiracles and tracheae, the writer has thus far found nothing

which would prohibit at least a limited functioning of these organs. That

air will pass through pores and tubes of smaller diameters than those of

the organs under discussion is now known, and many of the very minute

insects known to have a typical holopneustic type of tracheation have

openings and tracheae no larger than the reduced ones of the gilled Nympli-

ula caterpillars. Likewise if the minute tracheoles of the tracheal system

which are less than one micron in diameter can transport atmospheric

gases, failure of the reduced spiracles and connecting tracheae to function

would have to be due to some feature other than the structure of the tubes

themselves. That there would no difficulty in the ventilation of such a

system has recently been shown by Krogh (1920a; 1920b) since in small

forms diffusion alone will provide the necessary oxygen transportation,

although it may be assisted by respiratory movements of the animal, if

the latter are manifested.

In certain insects having apneustic tracheation the spiracles and con-

necting tracheae are said to be temporarily open at the time of molting.

This, however, does not account for the open character of the gilled larvae

of Nymphula since sections of specimens in various parts of the stadia

involved were studied and all yielded the same result.

There seem to be no reasons for assuming that open spiracles and open

connecting tracheae are necessarily inimical to larvae existing in water

since certain well known forms, as for example, Bellura melanopyga, have

complete sets of open spiracles, yet are related to the water in such a way

that most or all of these organs are submerged for long periods of time.

It is not unlikely that still other aquatic larvae, thought to have true

apneustic tracheation, will be found to possess morphologically open

spiracles.

What part these oj)en spiracles ])lay in the life ol the forms involved

is difficult, at present, to specify. As has been pointed out, the gilled

Nymphula larvae can pass extended periods of time out of water, at least

RESPIRATORY MfXHANISM IX LEPIDOPTERA 45

during the last larval stadium. This indicates functioning of the spiracles,

since it is unlikely that the requisite amount of oxygen could be secured

by cutaneous means alone, especially after the surface of the body became

dry. In regard to submergence, it is possible that a provision against

penetration of water into the tracheal system is afforded in the combina-

tion of structures present. The marginal crown of guarding spines or their

derivatives, if hydrofuge in character, may constitute an efficient protec-

tion. It is also possible that the well developed closing apparatus plays

some part in this connection.

General Considerations

From the point of view of the respiratory mechanisms involved, the

true aquatic Lepidoptera comprise a heterogeneous assemblage, including

those, on the one hand, which have made no morphological advance to-

wards the aquatic life, and those on the other hand, which manifest highly

developed morphological adaptations of an aquatic sort. These adapta-

tions involve, in the most complex type, the addition of structurally com-

plex gills, and the marked reduction in size of spiracles and connecting

tracheae. It should be noted that apparently no advantage has accrued

to the possessors of the complex adaptation since in all of the situations

examined by the writer the non-gilled larvae, having the unmodified

tracheal system of a terrestrial type, have had every appearance of being

as successful in the aquatic medium as their more specialized relatives,

often existing in identically the same environment and offering an interest-

ing parallel in the solution of the same problems by very different means.

There would seem to be a considerable advantage in the possession of over

400 gill-filaments and a set of reduced spiracles, particularly in those cases

where the size of the body proper is virtually that of the associated non-

gilled forms. As previously mentioned, the structure of the gills in Nymph-

ula is such that it seems almost inconceivable that they do not function

as true respiratory organs. In a recent paper. Fox (1921) claims to have

demonstrated that in a certain chironomid larva, oxygen is not taken up

by the ventral blood gills; that the anal gills take up less than the corre-

sponding area of the body-surface; and that most of the oxygen is received

through the body-wall in general. These surprising results require con-

firmation and at present need not be regarded as serious ground for ques-

tioning the function of the gills in other insects.

On the basis of structure alone it might appear that the small blood

gills of Cataclysta represent a more primitive stage in the development of

aquatic respiratory adaptation than that represented in the gilled larvae

of Nymphula. However, there is no indication that any of the Nymphula

species have ever had blood gills only. The evolution of these larval

46 PAUL S. WELCH

adaptations has apparently been a sporadic phenomenon with the extremes

sometimes occurring within the confines of a single genus.

Summary

1. Fundamentally, aquatic larvae of the genus Nymphula have retained

the original terrestrial type of body tracheation in practically unmodified

form. The tracheation of the gills has been superimposed upon the terres-

trial type with minimal change to the latter, and the non-gilled larval type

is doubtless the older one phylogenetically.

2. Gills in the aquatic Lepidoptera are all hollow outgrowths of the

body-wall, the cavity being in direct communication with the haemocoele.

All of the layers of the body-wall are represented but in reduced thickness,

maximum diminution appearing in the dermis.

3. In all of the gilled larvae of Nymphula examined, the gill-cavity con-

tains both an elaborate set of tracheae and tracheoles and a considerable

quantity of body fluid, thus constituting a combination tracheal-blood

gill. In the larvae of Cataclysta fulicalis the gills have no traces of tracheae

and are thus blood gills only.

4. In the non-gilled larvae of Nymphula and the gilled larvae of

Cataclysta fulicalis the tracheation is typically holopneustic, no reduction

of any significance appearing either in the spiracles or their connecting

tracheae.

5. In gilled larvae of Nymphula, a distinct reduction appears in the

spiracles and their connecting tracheae on segments 1, 4, 8, 9, 10, and 11,

those on segments 5, 6, and 7 being much larger and having undergone less

reduction.

6. Morphological and experimental studies on gilled Nymphula larvae

have shown that in spite of the striking reduction of some of the spiracles

and connecting tracheae, the tracheation is still holopneustic, all spiracles

and tracheae being morphologically open with nothing to indicate that

they are functionless. While gilled representatives of foreign species of

this genus have not been available, it is very probable that the statements

in the literature to the effect that they have a closed tracheal and spiracu-

lar system are in error, due to insufficient study.

7. The gilled larvae of Nymphula macidalis may live for extended

periods of time outside of water, even after the outer surface becomes dry

and the gill-filaments shriveled, indicating that respiration through the

spiracles is being accomplished.

8. Reduction of spiracles and possession of gills do not seem to be

necessarily correlated or coexistent since in Cataclysta fulicalis both gills

and an unreduced tracheal system are present.

9. In spite of the contrast between the gilled and the non-gilled species,

the former seem to have no advantage over the latter, at least in those

cases where both forms exist side by side in the same habitat.

RESPIRATORY MECHANISM IN LEPIDOPTERA 47

Literature Cited

Fox, H. M.

1921 Methods of Studying the Respiratory Exchange in Small Aquatic Organisms, with

Particular Reference to the Use of Flagellates as an Indicator for Oxygen Con-

sumption. Journ. Gen. Phj's., 3:565-573. 5 fig.

Hart, C. A.

1895 On the Entomology of the lUinios River and Adjacent Waters.

Bull. 111. State Lab. Nat. Hist., 4:149-273. 15 pi.

Krogh, A.

1920a Studien iiber Tracheenrespiration. II. liber Gasdiffusion in den Tracheen.

Pfliiger's Archiv ges. Phys. d. Menschen u. d. Tiere, 179:95-112. 5 fig.

1920b Studien iiber Tracheenrespiration. III. Die Kombination von mechanischer

Ventilation mit Gasdiffusion nach Versuchen an Dytiscuslar\'en.

Pfliiger's Archiv. ges. Phys. d. Menschen u. d. Tiere, 179:113-120. 2 fig.

Lloyd, J. T.

1914 Lepidopterous Larvae from Rapid Streams.

Journ. N. Y. Ent. Soc, 22:145-152. 2 pi.

1919 An Aquatic Dipterous Parasite, Ginglymyia acrirostris Towns., and .Additional

Notes on its Lepidopterous Host, Elophila fulicalis.

Journ. N. Y. Ent. Soc, 27:263-265. 1 pi.

Muttkowski, R. a.

1920. The Respiration of Aquatic Insects. A Collective Review.

Bull. Brook. Ent. Soc, 15:89-96, 131-141.

1921a Copper in Animals and Plants.

Science (N.S.), 53:453-454.

1921b Studies on the Respiration of Insects. I. The Gases and Respiratory Proteins of

Insect Blood.

Ann. Ent. Soc. Am., 14:150-156.

1921c Copper: Its Occurrence and Role in Insects and Other Animals.

Trans. Am. Micr. Soc, 40:144-157.

Portier, p.

191 1 Recherches Physiologiques sur les Insectes Aquatiques.

Arch. d. Zool. Exp., (5), 8:89-379. 4 pi. 67 fig.

Rebel, H.

1899 Zur Kenntnis der Respirationsorgane Wasserbewohnender Lepidopteren Larven.

Zool. Jahrb., abt. f. Syst., 18:1-26. 1 pi.

Rose, W. C. and Bodansky, M.

1920 Biochemical Studies on Marine Organisms. I. The Occurrence of Copper.

Journ. Biol. Chem., 44:99-112.

Welch, P. S.

1914 Habits of the Larva of Bellura melanopyga Grote (Lepidoptera)

Biol. Bull., 27:97-114. 1 pi.

1915 The Lepidoptera of the Douglas Lake Region, Northern Michigan.

Ent. News, 26:115-119.

1916 Contribution to the Biology of Certain Aquatic Lepidoptera.

Ann. Ant. Soc Am., 9:159-187. 3 pi.

1919. The Aquatic Adaptations of Pyrausta penitalis Grt. (Lepidoptera.)

Ann. Ent. Soc Am., 12:213-226.

Wesenberg-Lund, C.

1913 Wohnungen und Gehausebau der Siisswasserinsekten.

Fortschr. d. Naturwissensch. Forsch., 9:55-132. 59 fig.

48 PAUL S. WELCH

RESPIRATORY MECHANISM IN LEPIDOPTERA

49

4 (t 6

Plate X

Fig. 1. Longitudinal section through large spiracle and connecting trachea in larva of

Nymphula maculalis.

Fig. 2. Longitudinal section through spiracle and connecting trachea in larva of Cata-

clysta fulicalis.

Fig. 3. Longitudinal section through reduced spiracle and connecting trachea in larva

of Nymphula maculalis.

Fig. 4. Drawing of caustic potash preparation of large spiracle and connecting trachea

in larva of Nymphula maculalis.

Fig. 5. Transverse section of gill-filament in larva of Cataclysta fulicalis.

Fig. 6. Transverse section of gill-filament in larva of Nymphula maculalis.

50

PAUL S. WELCH

Plate XI

Fig. 7. Transverse section at ental end of connecting trachea for large spiracle in larva

of Nymphula maadalis, showing structure of closing apparatus.

Fig. 8. Transverse section at ental end of connecting trachea for spiracle in larva of

Nymphula oblikralis, showing structure of closing apparatus.

Fig. 9. Camera lucida drawing from living larva of N. maadalis showing tracheation of

gill-filament as it appears under low magnification. _

Fig. 10-11. Camera lucida drawings from living larva of N. macidalis showing distribu-

tion and arrangement of finer tracheoles of gill-filament as they appear under high magnifica-

tion.

DEPARTMENT OF METHODS, REVIEWS, ABSTRACTS,

AND BRIEFER ARTICLES

DICHROMATIC ILLUMINATION FOR THE MICROSCOPE

By

Leon Augustus Hausman, Ph.D.

The practise of using monochromatic light in photomicrographic work

is widespread. The advantages in the use of such light in conjunction with

ocular microscopic examination seem to be less appreciated; while the use

of dichromatic illumination, such as that described in this paper, the

writer believes to be new.

In using monochromatic light, i.e., light of a given color or wave-

length for microscopic examination, the purpose is three-fold: (1) to

increase the resolving power of the objective, (2) to secure greater con-

trast between different parts of the specimen, and (3) to afford relaxation

for the eyes. Abbe's formula for ascertaining the resolving power of an

objective is to multiply the numerical aperture of the objective by twice

the number per inch of the waves of the light employed. Hence it follows

that the shorter the wave length of the light, the greater the resolution

of the objective. This has an application of appreciable value in connec-

tion with the visibility and sharpness of focus of minute objects which

seem to lie just upon the border-line of vision. Moreover after working

long with white hght, green light affords a grateful relaxation to the eyes.

The microscope screen (S, Fig. 1) devised by the writer, has been

used with success both for monochromatic and dichromatic illumination.

With the former type of illumination the color filter is inserted in the slide

(CI, Figs. 1 and 2) and the light obtained from the arc-lamp (Al, Figs. 1

and 2).

For the examination of minute structures in the protozoan cell, and of

the pigment granules in the cortex of mammalian hairs, green or blue

illumination was found to be excellent. The former is more restful to the

eyes, especially when making protracted examinations. It is, moreover, of

greater luminosity, and hence permits of greater ease in focussing the

specimen. In the examination of protozoa the writer's practise is to focus

the object by the green light, and then exchange this for the blue. Violet

light was found unsatisfactory, because of its lack of luminosity. Certain

51

52

LEON AUGUSTUS HAUSMAN, PH.D.

Fig. 1

Assemblage of apparatus for dichromatic illumination. Microscope (M) in position

behind the screen (S), which bears slides (CiandC2) for supporting the color filters. Illumina-

tion of the object above and below is secured from the two arc lamps (Ai and A2).

Fig. 2

Diagrammatic view of apparatus for dichromatic illumination, from above. Ai and

A2 arc lamps, for transmitted and reflected light respectively; Bi and B2, plano-convex

condensers; the former to deliver practically parallel rays to mirror, the latter to illumine

object from above; Ci and C2, apertures for color filters; M, microscope. The screen (S)

is stippled.

DEPARTMENT OF METHODS 53

stained structures show up well with monochromatic light, particularly if

the color used be complementary to the one used as stain.

By employing a new type of illumination, i.e., dichromatic, it is possible

by means of the illumination alone to invest certain portions of the speci-

men with one color, and other portions with another, for the purpose of

bringing out thus by contrast the forms or relationships of the two. The

principle of dichromatic illumination is to illuminate the microscope field

and the transparent portions of the specimen with one color, by means

of transmitted light, and the more dense, or opaque portions of the speci-

men with another color, by means of reflected light, and to secure the

maximum contrast between the structures so illuminated by using comple-

mentary colors.

Such a result has been secured by means of the apparatus shewn in

Figs. 1 and 2 (whose letterings are similar). The microscope (M) is suffi-

ciently protected from all rays of light save those desired, by the screen

(S), which bear two slides (CI and C2). In these slides are apertures

wherein can be placed color filters. Filter CI delivers to the microscope

mirror light of one color for illuminating the field and the transparent

portions of the specimen, and filter C2 delivers light of another color which

is focussed upon the specimen from above. The transparent portions

of the specimen are therefore viewed by transmitted light of one color,

the opaque portions by reflected light of another color, and the translucent

portions by a combination of these two. Slide CI bearing its filter can

be moved laterally, and slide C2 vertically. The positions which the filters

can be made to occupy, together with the various positions of the micro-

scope within the screen, make it possible to secure light from any of the

angles useful in microscopic work. Illumination is furnished by two mov-

able arc-lamps, Al for delivering light to the mirror, and A2 for lighting

the specimen from above.

Within the microscope screen two movable plano-convex condensers

are used; one of long focus (Bl), for delivering to the mirror virtually

parallel rays; the other (B2) of short focus for condensing the light upon

the specimen from above.

With such an apparatus the writer has secured good results in the

examination of mammalian hairs, for the detection of delicate scalation.

With further experimentation the use of such lighting may possibly be

widened.

Color filters for use in photomicrographic work can be purchased and

used in the screen. Good filters can be made, however, in the laboratory

by developing out unexposed lantern slides, or dry plates, and then staining

the clear gelatin film with the desired colors. Care must be taken to secure

color filters that will allow only one color of light to pass through them,

i.e., they must furnish, as nearly as possible, monochromatic light. Some

54 LEON AUGUSTUS HAUSiMAK, PH.D.

of the stains, in aqueous solution, which have been recommended for this

purpose are: (1) Yellow — a saturated solution of picric acid. This

absorbs almost completely the blue and violet end of the spectrum.

(2) Green — methyl green. The absorption spectra varies with the depth

of the color. (3) Copper sulphate. This absorbs the red almost entirely.

Methyl blue is fairly good. (4) Red — Safranin.

The procedure in making color filters from lantern slide plates is to

develop and fix the unexposed plates and then to allow the gelatin film

to soften by placing Ihe plates in a bath of slightly warm water, say 80

degrees F. for a few minutes. They may then be removed to baths of the

different stains, and allowed to soak for an hour or so, or until the gelatin

is evenlv stained. It was found best to make up a series of saturated

aqueous solutions of the stains, and from these gradually to increase the

depth of color of the various baths until the desired depth was secured

in the gelatin film. The optimum for color filters is the greatest depth of

color which one can use and still secure sufficient luminosity for good focus.

The depth of the color of the filter giving the best results will depend upon

the brightness of the illuminant. This latter should give a strong, white

light. Such a light, with a spectrum very much like that of sunlight, can

be obtained from the electric arc.

A MODIFIED BARBER PIPETTE

The writer has been much inlerested in the various moditications of

the Barber pipette. One, which seems to be more simple than any here-

to-fore described, has been in use in these laboratories since 1916. It

seems worth while to describe it. The device consists of a bar A (side

view — figure 1, top view — figure 2) fashioned to fit partly around the

objective and fastened to it by the thumb screw B. The rod C passes

thru this bar and is held in position by the thumb-screw D. The bar E

is fastened to the rod C by a screw which also passes thru the spring F.

The part G is composed of a ring thru which a glass rod passes (the latter

held in position by the thumb screw H) and an angular piece with pivots

at the angle. There is a thumb screw I which controls the upward and

downward movement of the point of the pipette J. The camera bulb K

completes the apparatus. This device, minus the hollow tube and the

camera bulb, was devised l^y the late Dr. J. J. Wolfe for use in the picking

up of diatoms by adhesion. The hollow tube and camera bulb were added

by the writer for picking up copepods. This device later became very

useful in the segregation of living diatoms. A mechanical stage may be

used with the pipette. Altho the laboratory possesses a real Barber

pipette, this simpler apparatus is preferable when diatoms or larger or-

ganisms are to be segregated.

Bert Cunningilxm.

Trinity College,

Durham, N. C.

y::)

CLEANING SLIDES AND COVERS FOR DARK-FIELD

WORK

As particles of dirt, show with the same brilliancy as the bodies one

wishes to study, the slides and covers must be made very clean.

One of the most satisfactory ways of cleaning the slides is that suggested

by Stitt in his book on the blood etc., p. 235, 5th Ed. near the top. I have

found the modification here given very satisfactory:

(1) 5 grams of powdered Bon Ami are mixed with 100 cc of water and

thoroughly shaken up.

(2) New slides and covers are put into this mixture and well wetted,

then they are taken out and stood up on blotting paper to drain and dry.

The method answers well also for the final polishing of cover-glasses that

have been cleaned in acid-dichromate mixture, and for used slides that

have been cleaned in any approved method.

(3) Whenever one wants a slide or a cover one of those on which is

the dried Bon Ami is taken and wiped with a clean piece of gauze. It is

astonishing how quickly and well the cleaning can be done in this way.

Very few of them show any particles of dirt with the dark-field miscroscope.

(4) Cleaning the used slides and covers. — Hot water is allowed to flow

on the slide to wash off the oil, then the cover is removed and put into

cleaning mixture (Sulfuric acid and dichromate). The slide can usually be

well cleaned with the Bon Ami.

S. H. G.-^GE.

56

PROCEEDINGS OF THE AMERICAN MICROSCOPICAL

SOCIETY

Minutes of the Toronto Meeting

The 40th annual meeting of the American ^Microscopical Society was held in affiliation

with the American Association for the Advancement of Science at Toronto, Canada, Decem-

ber 29, 1921.

In the absence of the President, Frank Smith, and both Vice-Presidents, Professor

Albert M. Reese acted as Chairman.

The report of the Treasurer for the year 1921 was read by the Secretary and referred to

an Auditing Committee composed of Profs. R. J. Pool and R. H. Wolcott.

The report of the Custodian was read by the Secretary and referred to an .Auditing Com-

mittee composed of Messrs. Edw. Pennock and F. E. Ives.

The meeting voted to send congratulations to the Custodian, Mr. Magnus Pflaum on

the growth of the Spencer- Tolles Fund.

The Secretary presented a general report on the affairs of his office.

The following officers were nominated and elected for the constitutional periods: Presi-

dent, Dr. N. A. Cobb, Bureau of Plant Industry, Washington, D. C; Urst Vice-President,

Professor E. M. Gilbert, University of Wisconsin; Second Vice-President, Professor Z. P.

Metcalf, North Carolina State College of Agriculture and Engineering; Custodian, Mr.

Magnus Pflaum. Philadelphia, Pa.

Dr. B. H. Ransom, Bureau of Animal Industry, Professor Chancey Juday, University

of Wisconsin, and Professor George R. La Rue, University of Michigan, were chosen as the

elective members of the Executive Committee for 1922.

Adjourned.

P. S. Wfi.ch, Secretary.

Reports of the Treasurer and Custodian

Because of unavoidable delays, the reports of the Treasurer and the Custodian can not

appear until the April number.

57

>-^

TRANSACTIONS

OF THE

American

Microscopical Society

Organized 1878 Incorporated 1891

PUBLISHED QUARTERLY

BY THE SOCIETY

EDITED BY THE SECRETARY

PAUL S. WELCH

ANN ARBOR, MICHIGAN

VOLUME XLI

Number Two

Entered as Second-class Matter August 1?, 1918, at the Post-office at Menasha

Wisconsin, under Act of March 3, 1879. Acceptance for mailing at the

special rate of postage provided for in Section 11 03, of the

Act of October 3, 1917, authorized Oct. 21, 1918

ullip dnllpgiatp l^reaa

Geokge Banta Publishing Company

Menasha, Wisconsin

1922

TABLE OF CONTENTS

For Volume XLI, Number 2, April, 1922

On the Protozoa Parasitic in Frogs, with thirty-six figures, by R. Kudo 59

A New Suctorian from Woods Hole, with one plate, by F. M. Root 77

Department of Summaries

Ten Years of Heredity, with eight figures, by A. Franklin Shull 82

Department of Methods, Reviews, Abstracts, and Briefer Articles

A New Micro-slip, with one figure, by F. J. Myers 101

Killing, Staining and Mounting Parasitic Nematodes, by H. G. May 103

A New Locality for Spongilla wagneri Potts, by Frank Smith 106

Some Interesting Studies on Spider Anatomy, with one plate, bj' E. W. Roberts. . 107

Annual Report of the Treasurer 110

Custodian's Report for the Year 1921 Ill

^'1

TRANSACTIONS

OF

American Microscopical Society

(Published in Quarterly Instalments)

Vol. XLI APRIL, 1922 No. 2

ON THE PROTOZOA PARASITIC IN FROGS*

By

R. Kudo

Universitj' of Illinois

Probably no other animals have been for many year.s more favorite

objects of studies by zoologists than the frogs. The amphibians have been

examined by several Protozoologists and we know at present a consider-

able number of Protozoa of a great variety parasitic in frogs from various

parts of the world.

Numerous publications dealing with the protozoan parasites of frogs

have been issued by authors of several nationalities. Aside from the

papers by North American workers such as Ohlmacher (1893), Whinery

(1893), Gurley (1894), Stebbins (1904, 1905), Lewis and Williams (1905),

Metcalf (1909) and Swezy (1915, 1915a), a large majority are widely

scattered in various periodicals, and are not always easily referred to.

Undoubtedly this hardship concerning literature prevented the students in

Zoology from taking advantage of the material. If one possesses there-

fore brief accounts of the Protozoa commonly found in frogs, hundreds of

which are sacrificed yearly by students in Zoology and by special investi-

gators, one can utilize both material and time in carrying out observations

upon these interesting Protozoa.

The present paper is an attempt to meet this need. It deals with my

observations on the Protozoa parasitic in North American frogs which I

have examined during the last two years, together with the description of

methods of observation, and with brief review of and reference to the

works of the previous investigators on the subject.

The following six species are described in order:

1. Entamoeba ranarum from the intestine

2. Leptotheca ohlmacheri from the kidney

3. Haemogregarina sp. from the blood

*Contributions from the Zoological Laboratory of University of Illinois. No. 199.

59

60 R. KUDO

4. Trypanosoma rotatorium from the blood

5. Trypanosoma parvum nov. spec, from the blood

6. Opalina sp. from the intestine

I Entamoeba ranarum (Grassi) Dobell 1908

Habitat. — In the large intestine of Rana temporaria, R. esculent a, R.

clamitans and Bufo vulgaris. Dobell (1909) saw that about 23% of Rana

temporaria in Cambridge and Munich, were infected. I have seen a

number of amoebae whose characters agree on the whole with those de-

scribed by Dobell for Entamoeba ranarum m one out of 14 individuals of

Rana clamitans from New York in August of 1920. In Rana pipiens

which I have studied in 1920 and 1921 at Urbana, Illinois, I did not

observe any host individual that harbored the organism. This of course

does not mean its absence in a frog of this species, since I have not ex-

amined them as thoroughly as I did in the case of Rana clamitans.

Historical. — Lieberkiihn (1854) probably noticed the Amoeba in the

intestine of the frog which he studied. Grassi (1879) examined and

named it Amoeba ranarum. Dobell (1909) found an Amoeba in the frogs

of England and Germany, and studied them in detail. Quite recently,

the same author (Dobell, 1918) states that although the Amoeba resembles

closely morphologically to Entamoeba histolytica of human intestine, they

are distinct species. I have met with apparently the same Protozoon

but once, and could not carr}^ observation concerning its development.

Distribution. — Europe and North America.

Methods of observation. — A portion of the large intestine of a frog is

cut into small pieces in physiological salt solution on a cover-glass, made

an ordinary fresh preparation and observed. The organism may live for

several hours. The general appearance, changes in form of the body

through the formation of pseudopodia and the structure of the protoplasm

can be studied. To make permanent preparations, make smears on

slides or cover-glasses and fix them with hot sublimate-alcohol-acetic

mixture (2 parts of saturated aqueous solution of corrosive sublimate,

1 part of absolute alcohol and 5% of glacial acetic acid) for about 20

minutes. The smears are then immersed for about 15 minutes in a weak

iodine alcohol (50%) and then transferred into a plain alcohol to remove

the iodine. Staining with Delafield's haematoxylin, Heidenhain's iron

haematoxylin or Dobell's alcoholic haematcin, brings out satisfactory

results.

Morphology. — Amoeba of moderate size. When alive, the cytoplasin is

poorly differentiated into ectoplasm and endoplasm. Lobose pseudopodia

are actively formed at one time from any part of the body. The peripheral

portion of the cytoplasm is somewhat hyaline, while the main part of the

body is granulated and contains bacteria, yeasts and other particles

ON THE PROTOZOA IN FROGS

61

present in the host intestine. The nucleus is spherical and faintly visible in

living condition with an oil immersion objective. No contractile vacuole

is present. Dimensions vary from 15 to 40/i in the largest diameter.

When stained, the cytoplasm becomes highly vacuolated or reticulated.

The nucleus is spherical and usually contains a distinct karyosome.

Figs. 1 and 2. Entamocha ranarum. Fig. 1, a living individual. Fig. 2, an individual

stained with Delafield. .\ 1500.

Development. — According to Dobell, the cysts are found in the host

intestine in winter months. They are spherical, and measure 10 to 16/i

in diameter with a large nucleus. The nucleus divides twice producing four

daughter nuclei. Further changes are not known. Dobell suggests that

the cysts serve for the dissemination of the organism. The same author

(Dobell, 1918) recently found that although Entamoeba ranarum and E.

histolytica can hardly be distinguished morphologically from each other,

the cysts of the latter species when introduced into the intestine of tad-

poles did not undergo changes which take place in their proper habitat,

and concluded that these two forms should be held as different species.

II Leptotheca ohlmacheri (Gurley) Labbe 1899

Synonyms. — Chloromyxum (Sphaerospora) ohlmacheri Gurley 1893,

Leptotheca ranae Thelohan 1895 and Wardia ohlmacheri Kudo 1920.

Habitat. — In the kidney of Rana clamitans, R. pipiens and Bufo

lentiginosus. Out of 14 individuals of Rana clafnitans examined in New

York from July to September, 1920, six were infected by the parasite.

Out of 24 Rana pipiens bought from a Chicago biological supply store

and examined between November and December, 1920, ten were found

to be infected by the Myxosporidian. Thelohan (1895) named a Myxo-

sporidian which he saw in the kidneys of Rana esculenta and R. temporaria,

Leptotheca ranae. He has not given description nor figure for it, but I am

inclined to think this is probably identical with the American species.

62 R. KUDO

Historical. — The spores of the Myxosporidian were first found by

Ohlmacher (1893). Whinery (1893) also worked on them. Gurley

(1894) summarized the observations of the two authors. Thelohan

(1895) found Leptotheca ranae in France (?). No body seemed to have

worked on the organism until 1920 when I found the vegetative stages

and spores of a species what appeared to be identical with the present

form. I have studied its morphology and development, the result of

which will be stated elsewhere (Kudo, 1922).

Distribution. — North America and Europe (?).

Methods of observation. — When the infection is heavy, isolated spores

may be found in the cloaca of the host, but the kidney must be examined

for both spores and trophozoites. A small part of the kidney is cut into

small pieces on a slide in a drop of physiological salt solution and made

fresh preparation. In order to remove the fat globules that are usually

present in smears of kidneys, one drop of weak potassium hydrate solution

may be added to it. If any spores are present, they will be easily recog-

nized under a low power due to their peculiar appearance. If the infection

of the kidney is detected, hanging drop preparations or fresh preparations

with physiological solution should be made immediately and sealed with

melted parafilin. By using oil immersion objective, one can distinctly see

the detailed structures of the spores and trophozoites of various develop-

mental phases. To make permanent preparations, smears of variable

thickness should be made. In thinly made portion, one can see the

number and structure of the nuclei in well stretched trophozoites, while in

thickly made part, the shape, general appearance and arrangement of

nuclei and cytoplasm around them may be studied. Smears are well

fixed with sublimate-alcohol-acetic mixture. For staining, besides the

three methods stated for Entamoeba ranarum, Giemsa's method brings

out beautiful results. Section preparations should also be made in order

to observe the location of various developmental stages of parasites in the

host organ and the relation between the parasite and host body.

Morphology. — Fully grown trophozoites are usually rounded or oval

in form. Long conical pseudopodia are actively formed. Frequently

the trophozoites are completely rounded without any pseudopodia.

The body is colorless, granulated and extremely h}'aline. The cyto]>lasm

is indistinctly differentiated into endoplasm and ectoplasm. The cndo-

plasm is finely granualted and contains a vegetative nucleus, two develop-

ing spores and fat globules of variable size and number. The ectoplasm is

only distinctly visible where the pseudopodia are formed, the latter

being usually entirely composed of ecto])lasm. Before starting spore

formation the trophozoites multi])ly by active gemmation. In almost

all cases, disporous, rarely monosporous and more rarely trisporous.

ON THE PROTOZOA IN FROGS

63

Figs. 3 to 6. Trophozoites of Leplotheca ohlmacheri. Fig. 3, a trinucleate trophozoite

with a vegetative nucleus and two generative nuclei. A thin smear stained with Delafield.

Fig. 4, a young trophozoite in which four polar capsules are beine formed: fresh preparation.

Fig. 5, a thinly spread trophozoite with a vegetative nucleus and two developing spores.

Giemsa. Fig. 6, a rounded trophozoite with two mature spores: fresh preparation. All

X 2350.

Dimensions of fully grown disporous trophozoites are 30 by 20^t, 38 by

25/i, 40 by 20/z, etc. Each spore develops independently.

Development. — With regard to the interesting development of the

Myxosporidian the reader is referred to one of my papers (Kudo, 1922).

Morphology of the spore. — Oblong with its largest diameter standing

at right angles to the sutural plane. Anterior end is conspicuously

attenuated due to the thickening of the spore membrance at this point

while the posterior end is rounded. In lateral view, it is nearly circular

with a pointed anterior end. In an anterior end view, it is regularly

oblong. The spore membrane is moderately thick. Sutural ridge is well

marked, especially at the anterior end. The membrane is somewhat

irregularly striated. Three to seven fine striae run parallel to the sutural

64

R. KUDO

line on each valve and the remaining striae make somewhat similar angles

with the sutural line. The striae in lateral view run parallel to one

another except those on the posterior margin where a few make angles with

the former. The striae on each valve vary from 25 to 35 in number.

Figs. 7 to 13. Spores of Leptotheca ohlmacheri. Figs. 7, 8 and 9, the upper surface,

optical section and lower surface views of a normal fresh spore. Fig. 10, anterior end view of

a fresh spore. Fig. 11, an optical section of a fresh spore with two large sporoplasms. Fig.

12, a fresh abnormal spore showing two sporoplasms and two capsulogenous cells, each con-

taining a polar capsule. Fig. 13, a section through a spore showing the deepi)' stained polar

capsules and two uninucleate sporoplasms. Section: Delafield. Figs. 7 to 11, x 1500; Figs.

12 and 13, x 2350.

Two polar capsules usually equal in size, occupy the anterior portion of the

spore. The polar filament is coiled four to six times and is distinctly

visible in fresh condition. It can be extruded under the action of potas-

sium hydrate or mechanical pressure as I stated elsewhere (Kudo, 1918,

1921). Without staining the filament can be seen under a low magnifica-

tion. Two independent sporoplasms occupy the extracapsular cavity of

the spore, which condition is very rarely seen in Myxosporidia. They

appear homogeneous in fresh state. Staining reveals that each sporo-

plasm contains a single nucleus. Dimensions of fresh spores: sutural

diameter and thickness, 9.5 to 12/i, breadth, 13 to 14.5^i, diameter of polar

capsules 3.5 to 4.5 fx, length of extruded polar filament, 42 to 62 (x. Those

of stained spores: sutural diameter and thickness, 8.5 to 10 ju, l)readth,

9 to 12 /z, diameter of polar capsules 3 to 4 ^t.

Ill Ilaemogregarina .sp.

Our knowledge of haemogregarines is still in great confusion because

their development has not been studied except species occurring in reptiles.

The haemogregarine described here seems to agree with the following

ON THE PROTOZOA IN FROGS 65

species: Drepanidium magnum Grassi et Feletti 1891, Drepanidium krusei

Labbe 1892 and Karyolysus clamatae Stebbins 1905.

Habitat. — In the blood cell and plasma of Rana clamitans and Rana

pipiens. I have observed it quite frequently in the last named host

species. Quite frequently the frogs were found to harbor trypanpsomes

at the same time.

Historical. — While Lankesterella minima seems to have repeatedly been

studied by European authors (see for instance, Hintze, 1902), the present

form has been seen rarely. It seems to be this form that attracted the

attention of some North American investigators such as Langmann (1898-

1899) and Stebbins (1905). I have seen quite frequently the haemogre-

garine in frogs of New York and Illinois, but so far have not seen Lankes-

terella minima, the common European form.

Methods of observation. — Same as those stated for trypanosomes.

Morphology. — The haemogregarines found in the blood of frogs may

be spoken of under two types: intracorpuscular and extracellular. The

intracellular stage is cylindrical in shape, usually lying on one side of the

erythrocyte and displacing the nucleus to the other side. The posterior

end is usually folded up. In fresh preparation, the oblong nucleus with a

distinct membrane and usually a single karyosome is seen to occupy the

central portion of the body. The cytoplasm is homogeneous and contains

refractive granules of variable number scattered both in the anterior and

posterior regions of the body. Ordinarily there is no recognizable move-

ments of the body except at the time prior to the emergence from the

host cell in which the parasite is lodged. Around the body there is a thin

but distinct membrane. When stained, the oblong nucleus assumes two

kinds in appearance: one with eccentrically located karyosome and the

other with chromatin granules scattered evenly on the linin network. The

cytoplasm is highly reticulated and is always denser at the rounded end

than at the other end. The nucleus of the host cell seems to degenerate by

breaking down into a number of smaller irregular masses, and becomes

faintly stained, which condition indicates that the infection probably

causes some changes in the chemical nature of the nucleus of the host cell.

The host cell containing the haemogregarine becomes stretched and exhibits

variable shape and size. The number of parasites present in one host cell is

usually one, but frequently two are present, in which case the host cell

becomes greatly enlarged and deformed. The intracorpuscular forms

while under observation start to turn around in the host cell, and finally

breaks through the host cells. W^hether this is due to the pressure caused

by the cover-glass and by immersion objective or natural phenomenon

cannot be determined.

The extracorpuscular stage is gregarine-like in its appearance and

movements. The forms found in Rana pipiens and R. clamitans differ

66

R. KUDO

somewhat. The former is shorter and thicker and its anterior end is less

truncate with a nucleus situated near the anterior extremity, while the

latter is longer and thinner and its anterior end is more truncate with a

centrally located nucleus. I have not noticed the difference in living

condition, as their length and shape changed from time to time due to the

movements. The difference noted in the stained preparations may indi-

cate different circumstances in preparing them, although practically same

methods were used in both cases.

P'igs. 14 to 21. Ilaemogrcgar'nni sp. Kig. 14, a normal red blood corpuscle. Fig. 15, a

triniicleate erythrocyte containing an intracellular stage (the structure of one of the nuclei is

omitted). Figs. 16 to 18, three infected erythrocytes. Fig. 19, an erythrocyte containing

two parasites. Fig. 20, the parasite is just leaving the host cell. Fig. 21, an extracellular

giant form still covered with an envelope. Scluiudinn: Delafield. x 2100.

The free haemogregarines may be seen usually soon after the prepara-

tion was made, Jjut they increase in number in a few minutes. I have

often noticed the fact that when a fresh preparation of a small portion of

the lung of an infected frog was made, the number of e.xtracorpuscular forms

ON THE PROTOZOA IN FROGS

67

increased from 1 to 2 to from 12 to 20 in each field (compensation ocular 4

and apochromatic objective 8 mm.) in five minutes while under observation.

The body is rounded or truncate at the anterior extremity and tapers to an

attenuated posterior end. As was noted by many authors in several

species of haemogregarines, the posterior end of the animal is seen con-

nected with a thread-like structure sometimes measuring twice as long as

the body. It seems to me that it is a portion of the cytoplasm of the host

cell which was in direct contact with the parasite before the latter left it,

Figs. 22 to 29. ifaemogregarina sp. Figs. 22 to 24, e.xtracellular stages from the lung

capillaries of Rana pipiens. Fig. 25, an extracellular form from Rana clamitans. Figs. 26 and

27, erythrocytes of Rana pipiens with small forms. Figs. 28 and 29, erythrocytes with an

individual of Hacmogregarina sp. and a smaller form. Fig. 24, Giemsa; Fig. 27, Heidenhain;

the rest Delafield. x 2100.

and which became left behind as the animal moved forward. This view

also seems to be reasonable if one considers the fact that the thread-

like connection is most conspicuous soon after the parasite leaves the host

cell and disappears in a few minutes. The structure of the body in

68 R. KUDO

stained state is similar to that of the intracellular stage. The nucleus

assumes sometimes ring form. The average dimensions of forms found in

Rana pipiens: length 23.8 /z, largest breadth 3.6 /x: and of those found in

Rana damitans: length 27.6ju, largest breadth 2.4/i.

Development. — Ordinarily the dimensions of the parasites of both

phases described above are somewhat uniform. No stages of division

were noted either in the host cell or in free state. Smaller intracellular

stages such as shown in Figs. 26 to 29, are often observed. They seem to

occur always in the host cells. Oval with a flat or concave and a convex

side, they show at each end of the body one to three vacuole-like structures

in both fresh and stained conditions. These forms are always present with-

in the same host animal with the large forms described above. But no

intermediate stages between them have been seen, although a few forms

such as shown in Figs. 28 and 29 are noticed. Without infection experi-

ment, I cannot say whether they are only different stages in the develop-

ment of one and the same parasite or entirely different forms. Stebbins

(1904) considered the smaller form as a distinct species and named it

Haemogregarina cateshianae. The development of haemogregarines of

frogs has not yet been worked out, although Haemogregarina stepanowi

found an earnest worker in Reichenow (1910) who described interesting

observations.

IV Trypanosoma rotatorium (Mayer)

Synonyms. — Paramoecium loricalum Mayer 1843, Paramoccium cos-

tatum Mayer 1843, Amoeba rotatoria Mayer 1843, Trypanosoma sanguinis

Gruby 1843, Monas rotatoria Lieberkiihn 1870, Undulina ranarum Lan-

kester 1871, Paramoecioides costatus Grassi 1882.

Habitat. — In the blood of Rana esculenta, R. temporaria, R. damitans,

R. pipiens, R. castebiana, R. galamensis, ^. oxyrhynchus, R. mascarensis,

Rappia marmorata, Bufo vulgaris, Bufo- regular ia, Letodadylus ocellatus,

Hyla viridis and H. arborea. A number of host frogs whose specific names

were not determined by the original authors are excluded.

The trypanosomes are more numerous in the blood vessels of organs

such as liver and especially kidney than in the peripheral or heart blood.

Historical. — Since Gluge (1842) found the organism, several workers no-

ted and studied the flagellate, the chronological review of which is found in

Laveran and Mesnil (translated by Nabarro, 1907). Doflein (1910),

Lebedefif (1910) and Machado (1911) are more recent conributors to our

knowledge concerning this blood parasite of frogs.

Distribution. — Europe, Africa, Asia, South and North America.

Methods of observations. — The blood should be examined as soon as

it is taken from the frog. With a capillary pipette, draw the blood from the

frog heart. If it is taken aseptically, the Ijlood can be kc]U in sterile

ON THE PROTOZOA IN FROGS

69

condition in test tube with physiological solution and the trypanosomes

will live for several days. If one cannot observe the preparation soon

after the blood was taken, make temporary hanging drop preparations

which may be examined in two to six hours. The trypanosomes are the

largest ones known up to date, and its presence can be detected under a

low power, although one may have to examine several preparations of

the frog blood before finding one individual. When alive one can distinct-

ly see the undulating membrane and the characteristic wriggling move-

ment of the trypanosomes.

To make permanent smears, make ordinary blood smears and fix

with either sublimate-alcohol-acetic mixture or with absolute alcohol

(for 10 to 20 minutes). Staining with Delafield's haematoxylin, Heiden-

hain's iron haematoxylin or Giemsa's stain, will bring out morphological

details. The nucleus is sometimes hard to stain and prolonged staining

is needed to demonstrate its structure.

Morphology. — Polymorphic. Earlier observers held that the difference

in size and form among different individuals showed that of the specific

characters which view has however been abandoned by modern investiga-

Figs. 30 to 33. Trypanosoma rotaforium. Fig. 32, Delafield; rest Giemsa. x 1500.

tors. When the blood is examined soon after its removal from the frog

heart, one will see broad forms as well as slender ones mingled with inter-

mediate forms. After some time, some individuals become rounded.

The majority have more or less attenuated extremities. The form of the

70 R. KUDO

body changes constantly with the striking movements of the undulating

membrane. The flagellum which runs along the outer margin of the

undulating membrane is very frequently seen to extend beyond the

anterior end of the body. Its length varies, and may not be seen at all.

The blepharoplast and nucleus can hardly be seen in actively motile

individuals. The cytoplasm is granulated, may contain rounded spaces

especially near the posterior margin, and shows in many individuals

longitudinal striae.

When stained, the small oval or oblong blepharoplast is seen located

some distance from the posterior tip of the body. The flagellum seems to

take its origin a short distance from the blepharoplast. The nucleus is

located near the blepharoplast and on the same side of the body where the

latter is situated. It is rounded and shows its structure poorly with any

stain. It contains chromatin granules collected along the periphery. A

small karysome may sometimes be seen in the central region. The cyto-

plasm shows numerous small vacuoles in the posterior half of the body.

Dimensions vary considerably. Length, 44 to 70^i and breadth 10 to 35 ix.

Development. — Although artificial cultivation in vitro of Trypanosoma

rotatorium has successfully been carried out by Bouet, Doflein, Lebedeff

and Machado, we know practically nothing concerning its development in

the frog body. Some authors such as Laveran and Mesnil, Doflein, etc.,

have not seen trypanosomes undergoing division in the blood of host frog.

My own examination of numerous preparations also leads me to agree with

them on this point. On the other hand, Franca and Athias (1906), Dutton,

Todd and Tobey (1907) and Machado (1911) observed stages in division

in the smears of frog blood or in preparations of fresh blood of infected

frogs "kept aseptically at 72° to 89° F. for two or three days" (Dutton,

Todd and Tobey).

According to the observations of the last named three investigators,

the trypanosome after losing its flagellum, became rounded and underwent

active division, producing numerous small rounded organisms — "forty-one

cells were counted, though they were probably more." Each body became

ovoid, then pear-shaped, and from the more rounded end a flagellum

was produced. These young forms became active and free from the

outer covering of the original trypanosome. They divided rapidly by

splitting longitudinally increasing in number. They were herpetomonas-

like and remained in this condition until the preparations were discarded.

The authors studied further stages in stained smears, and stated that the

herpetomonas-like forms developed into inopinatum-Yike forms which were

also "found in fresh blood, in contradiction to the forms just described,

which were found in kept blood alone."

Machado describes stages in division of the trypanosomes in frog.

From his statement, it is not clear whether he found these stages in the

ON THE PROTOZOA IN FROGS 71

blood or kidney. Figs. 14 to 19 and 36 to 40 given by Machado are

rather isolated from others and hard to be reasonably connected with the

other stages which he figured. I have not had chance of examining

Leptodactylus ocellatus myself, but comparison of the above mentioned

Machado's figures with the vegetative stages of a Myxosporidian, Leptolhe-

ca ohhnacheri which is described very briefly in this paper and in details in

the other paper (Kudo, 1922) and which is not uncommon parasite of the

kidneys of Rana clamitans, R. pipiens and Bujo lentiginosus of the United

States, leads me to think that the frogs studied by Machado were probably

infected by the Myxosporidian or an allied form besides the trypanosomes.

Machado states that trypanosomes were abundantly seen in the kidney

of the host which fact I also noted. He seems to have mixed the stages

of development of a Myxosporidian with those of the trypanosomes.

Judging from the trypanosomes of fishes and reptiles, and Trypan-

osoma inopinatum, another member of the genus parasitic in Rana escidenta

of Algeria, the present species seems to undergo changes in the body

of blood sucking invertebrates. Fuller accounts of the life history of the

trypanosome awaits future investigations.

V Trypanosoma parvum nov. spec.

Habitat. — In the blood of Rana clamitans. Fourteen specimens were

examined between July and September, 1920. In one of them a fairly

heavy infection of a trypanosome was noticed. Five to eight active indi-

viduals were recognized in every field (compensation ocular 4 and apo-

chromatic objective 8 mm.). The frog also harbored Trypanosoma rota-

torium in small number (one individual in every two other field under the

same combination as noted above), but no haemogregarine was found.

I have not seen it since that time, although I have examined about four

dozens of Rana pipens which were purchased from a Chicago biological

supply store.

Methods of observations. — Same as Trypanosoma rotatorium. For

demonstration the unusually long flagellum, Fontana's staining was used

with satisfactory result.

Morphology. — When alive, the movements can be distinguished into

two types: travelling and wriggling movements, of which the first is

prominent. The active wriggling movements remind one of those of Try-

panosoma lewisi. The undulating membrane is fairly well developed.

The nucleus and belpharoplast are faintly visible, while the relatively

long flagellum can distinctly be seen with an oil immersion objective.

The cytoplasm contains frequently small rounded clear spaces, and is

more or less vacuolated at the posterior portion.

When stained, one finds in them structures typical to a trypanosome.

The body is spindle-shaped usually being curved in an arch or S. The

•72

E. KUDO

posterior end is ordinarily attenuated and ends in a blunt point, while the

anterior extremity is more sharply pointed. The cytoplasm is usually

dense from the anterior part to the middle region of the body, while a

clear area is frequently seen ju^t posterior to the nucleus, either close to or

Figs. 34 and 35. Trypanosoma parvum nov. spec. Fig. 34, Giemsa; Fig. 35, Delafield.

X 3300.

somewhat separated from it. The posterior portion is more or less

vacuolated as was noted in living specimens. The blepharoplast is located

very close to the posterior tip of the body. It is relatively large, and

rounded or oblong in shape. The flagellum that borders the outer margin

of the undulating membrane does not seem to take its origin directly in the

blepharoplast, but arises from a point inconspicuously marked at some

distance from the latter. The free portion of the flagellum reaches \S n

in length, though its length varies most widely. The nucleus is rather

large, and is located between the middle and anterior third of the body.

It is spherical or oval. In Giemsa stained smears, the peripheral portion

stains very deeply, while the central portion is occupied by a few linin

threads. A karyosome may sometimes be seen eccentrically located.

Dividing forms were not seen. The trypanosomes are strikingly

uniform in size, showing little variation in size and general shape, except the

length of the flagellum. Measurements of two hundred specimens in

smears fixed with sublimate-alcohol-acetic mixture and stained with

Giemsa's solution are as follows: length of body, exclusive the flagellum,

11 to 14 n, largest breadth including the undulating membrane, 1.2 to

1.9ju, length of free portion of the flagellum 5 to 15 /i.

ON THE PROTOZOA IN FROGS 73

Of all the trypanosomes of amphibians known up to date Trypanosoma

inopinatum Sergent et Sergent, 1904, resembles closely to the form just

stated. These two forms resemble each other in the dimensions and

general resemblance to Trypanosoma lewisi. There are however some

differences in the location of blepharoplast, the structure of cytoplasm and

the general form of the stained individuals which shows the activity of the

two forms is not same. The blepharoplast is located more closely to the

posterior tip in this form than in Algerian form. The breadth of the

American form is 1.2 to 1.9 n, while that of Trypanosoma inopinatum is

3 II. The cytoplasm of the present form is vacuolated at the posterior por-

tion of the body, while the Algerian form, according to Sergent and Ser-

gent's figures, is uniformly granulated. Furthermore the activity of the

two forms appears to be quite different. In the forms I have studied

the body shows an arch or S shape in stained smears, while Sergent and

Sergent figure more or less straight form, thus indicating probable differ-

ence in their activity when alive. Consequently these two forms should

better be separated from each other by different specific names, until I am

able to compare the preparations of them.

Since the cultivation of Trypanosoma rotatorium in vitro has been

attempted by Lewis and Williams (1905), the fact that the trypapnosomes

undergo division in the culture media resulting in the formation of small

spindle-shaped bodies resembling in appearance to Herpetomonas or

Crithidia, became known. But in no case, a structure typical to a try-

panosome was noted among these small forms. At our present state of

knowledge concerning trypanosomes, it is proper for us to consider the

extremely small trypanosome described above as independent from Try-

panosoma rotatorium. As it is morphologically distinguishable from a

closely allied form, Trypanosoma inopinatum, I propose to name it provi-

sionally Trypanosoma parvum nov. spec.

Parasitic flagellates in the intestine

Number of parasitic flagellates have been described in the intestine of

frogs. The reader is referred to Dobell (1909) and Swezy (1915, 1915a)

concerning them,

VI Opalina sp.

The Opalinas described here seem to be identical with Opalina ranarum

Purkinje et Valentin 1835.

Habitat. — In the rectum of Rana clamitans and R. pipiens.

Historical. — A complete chronological review of works on Opalinas

will be found in Metcalf (1909).

Methods of observations. — The rectum of frog is placed in a small

watch glass and opened in physiological salt solution under dissecting

microscope. When the preparation is made, the Protozoon will be seen

actively moving. In order to retard the active movements, a drop of

74

R. KUDO

two of cherry gum solution may be added. The ciliary movements and

the structure of the body can easily be studied. For permanent prepara-

tions, follow the methods stated for Entamoeba ranarum.

Morphology. — The body is broadly oval, with blunt anterior and more

rounded posterior extremities. One side is convex, while the opposite side

exhibits a shallow depression at the middle part. The body is highly

flattened. Parallel rows of cilia run obliquely. The body is covered with

cilia of uniform length. The protoplasm is sharply differentiated into

Fig. 36. Opalina sp. Delaficld. .\ portion of the body is shown in detail. .\ 400,

ectoplasm and endoplasm. The ectoplasm is hyaline near the pellicle,

but is alveolated near the endoplasm. The latter is granulated in living

individuals, l^ut when stained with Delafield's haematoxylin, it shows a

vacuolation. The endoplasm contains a large number of nuclei of uniform

size. Cytostome or cytpygc is not observed. Dimensions: length 130

ON THE PROTOZOA IN FROGS 75

to 200 /i, breadth 50 to 120 fi. Occasionally large form reaches 500 /x in

length.

Development. — The Protozoon divides in the intestine of the frog,

stages of which are commonly seen in the rectum in the summer. I have

not studied a new infection of a host frog. According to Neresheimer

(1907), Opalina rafiartim divides successively in the rectum of host frog in

the spring, and produces numerous small individuals, each containing a few

nuclei. They encyst by producing a hyaline and resistant membrane

around them. The cysts come out of the host body with fecal matters

and remain on the bottom of the water. When the young tadpoles

swallow the cysts, the contents of the latter leave the membrane in the

rectum of the new host. The free young opalinas become differentiated

into gametes by division and after fusion form zygotes. The zygotes

grow into adult ones as the tadpoles metamorphose themselves into adult

frogs.

Summary

1) The main object of the present paper is to furnish a brief account

of Protozoa parasitic in common North American frogs for general students

in Zoology.

2) The occurrence of Entamoeba ranarum in Rana clamitans is stated.

3) A Myxosporidian, Lcptotheca olilmachcri is studied in the kidneys

of Rana clamitans and R. pipiens.

4) Trypanosoma rotatormm of Rana clamitans and R. pipiens is studied.

5) Haemogregarina sp. in Rana clamitans and R. pipiens is studied.

6) A new trypanosome, Trypanosoma porvum is described from Rana

clamitans.

7) Opalina sp. from Rana clamitans and R. pipiens is studied.

8) Methods of observation and brief review of previous works for

each of these forms are given.

Bibliography

The papers marked with an asterisk contain the summary of and reference to the works

of previous investigators on the subject.

^ Enlamoeha ranarum

DOBELL, C. C.

*1909 Researches on the intestinal Protozoa of frogs and toads. Quart. Jour. Micros.

Sc, 5.1:201-276, 4 pi. and 1 textfig.

1918 Are Entamoeba htslolytica and Entamoeba ranarum the same species? An experi-

mental study. Parasit., 10:294-310.

„ Leplothcca ohlmacherl

Kudo, R. ^

*1920 Studies on Myxosporidia. A Synopsis of Genera and Species of Myxosporidia.

111. Biol. Monogr., 5:243-503, 25 pi. and 2 textfig.

1921 On the nature of structures characteristic of Cnidosporidian spores. Trans.

Micro. Soc, 40:60-74.

1922 On the morphology and life history of a Myxosporidian, Lcptotheca ohlmacheri,

parasitic in Rana clamitans and Rana pipiens. Parasitology, 14, no. 2.

76 R. KUDO

Ha emogrega rhics

DuTTOx, J. E., J. L. Todd and E. X. Tobey.

1907 Concerning certain Protozoa observed in Africa. II. Ann. Trop. Med. Paras.,

1:287-370, 13 pi. and 34 textfig.

Franca, C.

1917 Sur la classification des hemosporidies. Journ. Sc. Mate. Fis. Nat. Acad. Sci.

Lisboa, 3 ser. 41 pp.

HlXTZE, R.

1902 Lebensweise und Entwicklung von Lankeslcrella minima (Chaussat). Zool. Jahrb.

Abt. Anat., 15:693-730, 1 pi.

Stebbixs, Jr., J. H.

1904 Upon the occurrence of Haemosporidia in the blood of Rana catesbiana, with an

account of their probable life history. Trans. Amer. Micr. Soc, 25:55-61, 2 pi.

1905 On the occurrence of a large sized parasite of the Karysolysus order in the blood of

Rana cJamata. Centr. Bakter. I Abt. (Orig.) 38:315-318, 2 pi.

Billet .\. Trypanosomcs

1904 Sur le Trypanosoma inopinatum de la grenouille verte d'Algerie et sa relation

possible avec les Drepanidium. C. R. soc. bioL, 57:161-164, 16 textfig.

Brumpt, E.

1906 Role pathogene et mode de transmission du Trypanosoma inopinatum Ed. et Et.

Sergent. Mode d'inoculation d'autres trj^anosomes. C. R. soc. biol.,

61:167-169.

DOFLEIX, F.

1910 Studien zur Xaturgeschichte der Protozoen. VI. Experimentelle Studien iiber

die Trypanosomen des Frosches. Arch. Protist., 19:207-231, 3 pi. and 1

textfig.

DuTTOx, J. E., J. L. Todd and E. N. Tobey (see haemogregarines).

Laverax, a. and E. ;Mesxil (translated and revised by Nabarro).

*1907 Tr}-panosomes and trj'panosomiases. Chicago. 538 pp., 1 pi. and 8 textfig.

Lebedeff, a.

1910 Ueber Trypanosoma rotatorium Gruby. Festschr. 60sten Geburts. Richard

Hertwigs, 1:397-436, 2 pi., 9 textfig.

]Machado, a.

1911 Zytologische Untersuchungen iiber Trypanosoma rolalorium Gruby. Mem. Inst.

Oswaldo Cruz, 3:108-135, 2 pi.

Sergext, Ed. et Et.

1904 Sur un trypanosome nouveau, parasite de la grenouille verte. C. R. soc. biol.,

56:123-124, 1 textfig.

Dobell, C. C. Intestinal flagellates

*1909 (sec Entamoeba ranarum).

SWEZY, O.

1915 Binary and multiple fission in Hexamitus. Uni. Calif. Publi. in Zoology, 16:

71-88, 3 pi.

1915a On a new trichomonad flagellate, Trichomitus parvus, from the intestine of .Amphi-

bians. Uni. Calif. Publi. in ZoologA-, 16:89-94, 1 pi.

Metcalf, :M. M. Opalinac

*1909 Opalina. Its anatomy and reproduction, with a description of infection experi-

ments and a chronological review of the literature. Arch. Protist., 13. 181

pp., 15 pi. and 15 textfig.

Nereshelmer, E.

1907 Die Fortpflanzung der Opalinen. Arch. Protist., Suppl. 1:1-42, 3 pi. and 2 textfig.

A NEW SUCTORIAN FROM WOODS HOLE

By

Francis Metcalf Root, Ph.D.

Department of Medical Zoology, School of Hygiene and Public Health,

The Johns Hopkins University

The aberrant group of the Infusoria known as the Suctoria have been

but little studied in the United States. In Europe, however, the mono-

graphic work of Sand and, more recently, of Collin have made them well

known.

In 1914 I published some notes on the reproduction and food reactions

of a fresh- water species of the genus Podophrya which appeared in hay-

infusions at Baltimore. Through this work I became interested in this

peculiar group of the protozoa and during the summers of 1916 and 1917,

while at Woods Hole, Mass., I collected and studied five species of Suctoria

which attached themselves to the stalks of the common hydroids, Obelia

commissuralis and Obelia geniculata.

The four previously described species found were as follows:

Ephelota coronata (Wright).

Acineta tuberosa Ehrenberg.

Paracineta livadiana (Mereschowsky) — This is probably the species

referred to by Calkins (1901) as "Acineta divisa Fraipont."

Ophryodendron abe itinum Claparede et Lachmann.

Besides these four species another was present which was obviously new

and extremely remarkable, showing decided resemblances to Acinetopsis

rara Robin.

The genus Acinetopsis was established by Robin (1879) to contain a sin-

gle species, Acinetopsis rara, characterized by the presence of a stalked

theca with free apical margin (a "coque" in the terminology of Collin

[1912 page 117] and by the presence of a single extensile, flexible tentacle in

the center of the apical surface of the body. This species has never been

reported by any other observer. Martin (1909) attempted to identify

Robin's Acinetopsis rara with one stage in the life history of his new

species, Tachyblaston ephelotensis. This identification has not been

accepted by Collin in his monograph of the Suctoria, and will hardly be

agreed to by anyone who will take the trouble to compare Martin's figures

with the original figure of Robin, as reprinted in the Journal of the Royal

Microscopical Society (1880). More recently Collin (1909, 1912) has

described a new species under the name of Acinetopsis campanuliformis.

This species is described as having a bell-shaped "loge" or closed theca

77

78 FRANCIS METCALF ROOT, PH.D.

and bears six flexible tentacles. Collin seems to have overlooked the

statement of Robin that Acinetopsis rara has a theca with free margin,

and has classified Acinetopsis as a genus with closed theca. If this differ-

ence be really of generic importance, Collin's Acinetopsis campanuliformis

must be placed in a new genus.

The species which I found at Woods Hole so closely resembles Acine-

topsis rara at one stage of its life-history that I am not willing to erect a

new genus for it, in spite of the fact that in this new species I find that the

flexible extensile organ proves to be a seizing organ, analogous to the

pointed tentacles of Ephelota and without any suctorial function. To

this new species I have given the name of "tentaculata," because in its

adult condition it is provided with true sucking tentacles as well as with

the elongated "probosces" which I consider characteristic of the genus.

Description of Acinetopsis tentaculata n. sp.

Protoplasmic body enclosed in a flattened, cup-shaped theca with a free

apical margin, borne on a slender stalk a little longer than the theca itself.

(See Figure 1). Body irregularly flattened-ovoid in shape, bearing on the

apical face one or two probosces. Each proboscis is a very mobile organ,

which can be bent in any direction, extended until it is a mere thread twice

the length of all the rest of the organism, or retracted until it is less than

half the length of the protoplasmic body alone. Structurally, the proboscis

consists of a homogeneous central strand about whose outer surface is

wound spirally a ribbon of protoplasm evidently contractile in nature.

Each proboscis is tipped with a large, highly refractive globule of adhesive

and viscous character. That the probosces are firmly anchored to the

body is self-evident from the observations given later regarding their use

in feeding. As is shown in Figure 2, this attachment is well below the

surface of the body, but I have not been able to make out the details.

Besides these probosces the apical face of the body also bears about

twenty or thirty short stout tentacles of the familiar capitate type, these

being distributed in two groups, one on either side of the insertion of the

probosces.

The macronucleus has irregular outlines, but is in general roughly

ovoid. One or more small spherical micronuclei are present. A single

contractile vacuole is located near the base of the body.

The measurements of a typical mature specimen were as follows:

Protoplasmic body — 138 microns long, 100 microns wide, 73 microns thick.

Theca — 187 microns long, 105 microns wide.

Stalk — 287 microns long.

Proboscis when fully extended — about 500 microns long.

A NEW SUCTORIAN FROM WOODS HOLE 79

Feeding Habits

As far as I have been able to observe, Acinetopsis tentaculata feeds only

on Ephelota coronata, but it attacks this common species with voracity.

The probosces are extended and moved about until the globule at the

tip of one of them comes in contact with the body of an Ephelota. It

seems to secure a firm attachment at once and in a few cases where the

Ephelota was very firmly attached to its stalk I have seen the globule

elongate and finally pull in two rather than release its victim. As soon

as the attachment is secured the proboscis contracts strongly, drawing the

body of the Ephelota within reach of the short sucking tentacles of its

captor. Sometimes the stem of the Ephelota is long enough so that this

can be accomplished by merely bending it. More often the attachment

between the body of the Ephelota and its stem must be broken by a rapidly

repeated series of jerks due to sudden contractions of the proboscis. When

the Ephelota finally comes within reach it is firmly seized by the sucking

tentacles and its internal protoplasm sucked out at leisure. One of the

difficulties in the study of this species was the long search which was always

necessary to find an individual whose body was not almost entirely con-

cealed by several Ephelotas in process of being sucked dry.

Reproduction

The actual escape of the embryo was not observed, but Figure 3 shows

plainly that reproduction occurs by the same process of simple (i. e. not mul-

tiple) internal budding which is characteristic of all the genera of the

family Acinetidae. Nor have I observed the entire series of stages in the

attachment and growth of the free-swimming ciliated embryo. From the

young individuals I have found in nature, it seems probable that after

attachment and the formation of a stalk and theca, a single proboscis is

first formed (Figure 4). This condition is structurally a perfect duplicate

of Robin's Acinetopsis rara. Slightly larger individuals still show only a

single proboscis but have also formed a number of true tentacles (Figure 5).

And finally, in mature individuals, we find the two probosces characteristic

of the species.

There is a great temptation here to suggest that these growth stages may

reflect something of the course of evolution in this genus. It seems quite

probable, for example, that the second proboscis, the last organ acquired

by the individual, is a recent evolutionary acquisition of the species. It

would probably be going too far to intimate that the stage with a single

proboscis and no tentacles harks back to some ancestor in which the

proboscis was a true sucking tentacle and no other organs for capturing

prey were necessary. However, this must be the condition which obtained

in Robin's Acinetopsis rara, unless he overlooked the inconspicuous

tentacles or was dealing only with a growth stage.

80 francis metcalf root, ph.d.

Summary

Acinetopsis tentaculata n. sp. is described and figured. It is charac-

terized by having a stalked "coque" or theca with free apical margin,

and by the presence of one or two extensile seizing organs or probosces as

well as suctorial tentacles. It feeds on Ephelota, seizing them and drawing

them within reach of its tentacles by means of the probosces. It repro-

duces by internal budding, forming free-swimming ciliated embryos which

settle down and gradually grow into the adult form.

Literature Cited

Calkins, G. N.

1901 Marine Protozoa from Woods Hole. U. S. Fish Comm. Bull. Vol. 21, pp. 413-468.

Collin, B.

1909 Diagnoses preliminaires d'Acinetiens nouveaux ou mal connus. C. R. Acad.

sc. Paris. May 24, 1909.

1912 Etude monographique sur les Acinetiens. II. Morphologie, Physiologic, Syste-

matique. Arch. Zool. Exp. et Gen. Vol. 51, pp. 1-457.

Martin, C. H.

1909 Some observations on Acinetaria. II. The life cycle of Tachyblaston epheloten-

sis. Quart. Journ. Micr. Sci. Vol. 53.

Robin, C.

1879 Memoire sur la structure et la reproduction de quelques Infusoires. Journ.

Anat. et Physiologie. Vol. 15. (Reviewed with illustrations in Journ. Roy.

Micr. Soc. 1880.)

Root, F. M.

1914 Reproduction and reactions to food in the Suctorian, Podophrya coUini n. sp.

Arch, fiir Protistenkunde. Vol. 35, pp. 164-196.

Description of Plate

All figures are camera drawings from specimens killed in Dubosque's alcoholic modifica-

tion of Bouin's Fluid and stained with picric acid haematoxylin.

Figure 1. Acinetopsis tentaculata. Side view of entire mature specimen with one

proboscis partly extended and the other nearly retracted, x 185.

Figure 2. Acinetopsis tentaculata. Front view of body and theca of mature specimen.

Note deep attachment of probosces and arrangement of tentacles in two groups, x 350.

Figure 3. Acinetopsis tentaculata. Front view of an individual in which the macronu-

cleus is just dividing to form the macronucleus of an internal bud. x 350.

Figure 4. Acinetopsis tentaculata. Young form with a single proboscis and no ten-

tacles. X 350.

Figure 5. Acinetopsis tentaculata. Young form with a single proboscis and a few

tentacles, x 350.

A NEW SUCTORIAN FROM WOODS HOLE

81

DEPARTMENT OF SUMMARIES

DEVOTED TO DIGESTS OF PROGRESS IN BIOLOGY

TEN YEARS OF HEREDITY^

By

A. Franklin Shull

University of Michigan

Though no one is likely to be misled by my subject into supposing that

the laws of heredity have been in operation only a decade, it may not be

universally appreciated that heredity is one of the oldest of biological

phenomena. It is at least as old as, probably older than, organic evolution

of which we have long been accustomed to speak in terms of millions of

years. For, when the first living thing, if ever there was such a being,

gave rise to a second, by reproduction, this second Hving thing was either

like its parent, or different from it. If like its parent, heredity had begun.

If different from its parent, it was almost certainly different in only one

or a few respects, but Hke the parent in the rest, in which case both

heredity and evolution were in operation.

The stipulated ten years to which my title refers are not, however,

an arbitrary limit set for the purpose of relieving me of the necessity of

covering the whole of a very large subject. They are a period which,

in the development of knowledge of heredity, is naturally marked off from

the numerous decades that precede. Those of you who possess a little

knowledge of heredity, to whom the name of Gregor Mendel has a fasci-

natingly familiar sound, and in whose memory lingers the date of 1900 in

which year the famous Austrian monk's long hidden experiments were

again brought to light, may wonder why I should wish to describe the

developments of but the latter half of the period since that rediscovery.

It is true that the great interest aroused by the verification of Mendel's

Law, with the multiplication of experimental work which was induced

by it, was a necessary precursor of the events with which I propose to deal.

But about 1910 there began a chain of discoveries, which have followed one

another in unbroken series to the present time, and which have led to a

conception of the operations of heredity of a degree of complexity, and

withal of harmony, which even the most sanguine twenty years ago would

not have ventured to predict.

' This lecture, delivered before the Graduate Club of the University of Michigan, was

designed to present to persons without biological training, not a r^sum^ of all important

work in heredity in the period referred to, but the point of farthest advance and the principal

work leading to it.

82

department of summaries 83

Former Lack of Analysis

Heredity had long been discussed in terms of averages. In popular

discourse it was always so, children were replicas of their mothers, or were

chips out of the old block, or the son of a Cholmondeley was the image of

a Jones. The ensemble of characters was considered. Even those who

were professionally engaged in the study of heredity lumped together many

things now known to be partially or wholly distinct from one another,

regarding them as a single trait. Stature, obviously made up of many

elements, was treated as a single characteristic. Intelligence, likewise

compound, was studied as if simple.

There was not wanting, it is true, even among the laity, an analytic

tendency. A youth would have his father's mouth, his mother's eyes, his

grandfather's complexion. But it was not until the emergence of Men-

del's work in 1900, and the multiplication of investigations consequent

upon that event, that it was realized to what extent inherited traits may be

separated from one another as distinct and independent units. Eyes were

inherited independently of hair, hair color independently of hair form,

color independently of distribution of color, whether uniform or in patches.

Unit characters became distinctly vogue. Anyone who could utter the

magic expression "unit characters" and speak the name of Mendel with

his first name and title, had thereby established his right to be regarded

as a thoroughly modern geneticist.

Difficulties of the New Conception

All this development raised in the biological mind certain difficulties.

When it was inquired how all these unit characters were manipulated

independently of one another, there were obstacles — that is, when the

offered explanation passed a certain point. To speak intelligibly of these

difficulties it will be necessary to refer briefly to a few elementary facts of

biology.

All organisms are composed of units of structure called cells. These

cells regularly contain, as part of their structure, a rounded body, the

nucleus, which stains deeply in most dyes and which is therefore conspicu-

ous in most common microscopic preparations. The size of an organism is

increased usually by the multiplication of cells, which is accomplished by

the division of the cells already present. In the process of division, the

cells develop a complicated figure in which the highly staining material of

the nucleus is resolved into a number of distinct bodies called chromo-

somes. As the division is completed, the chromosomes lose their distinct

form, producing a nucleus in which separate bodies are no longer visible;

but at the next division the chromosomes appear again, in the same number

as in the previous division. This number is in general constant in all

cells of the same individual, and, barring some differences between the

34 A. FRANKLIN SHULL

sexes, is constant for all members of the same species. Moreover, in

animals in which the chromosomes are not all of the same size or shape,

each dividing cell reveals the same number of chromosomes of each shape or

size.

Since all organisms are composed of cells, the phenomena of heredity

must in some way be traceable to cells. The constancy of occurrence of the

nucleus, and of a given number of chromosomes in the nucleus, early gave

rise to a suspicion which later, on a foundation of fact, ripened into a

conviction, that in these structures is the mechanism through which

heredity is governed. If it were assumed that the factors of heredity were

contained in the chromosomes, many things would be explained. Refer-

ence will be made now to only one of these things.

One of the new features of discussion of heredity was the attention

devoted to unit characters. How were these characters operated as units,

independently of one another? Chromosomes provided the answer. It

must be understood that in all the higher animals and plants, no parent

contributes all of its chromosomes to any one offspring, hut only half of these

chromosomes. In the development of the germ cells a peculiar cell division

called the reduction division takes place in which the chromosomes separate

into two groups, one group being enclosed in the one daughter cell, the other

group in the other cell.

Chromosomes and Recombination of Characters

In the composition of these groups of chromosomes, there is a wide

range of different possibilities. In some cases, the chromosomes may be

Figure 1. Diagram of a cell in which the chromosomes a.rc capable of arrangement in

pairs, the two chromosomes of each pair being precisely alike in all respects. Such a cell,

in maturation, divides into two cells, each with half the number of chromosomes, and each

exactly like the other in all hereditary factors. The sliapes of the chromosomes are not

actual, but only a diagrammatic representation of the likenesses and differences in their

hereditary composition.

DEPARTMENT OF SUMMARIES 85

capable of assortment in pairs, as in figure 1 ; that is, there are two chromo-

somes in each cell that are exactly alike, two other chromosomes that are

exactly alike but different from the first pair, and so on. In such a case,

by separating the members of each pair, two groups of chromosomes may

be made up which are identical. In such a case, therefore, tivo germ cells

with exactly the same hereditary possibilities are produced, and the parent may

contribute precisely the same hereditary traits to every one of its progeny.

Moreover, it can transmit to each of its offspring every hereditary trait of

which it is possessed.

In other individuals, on the contrary, every chromosome of a cell may

differ in one or more respects from every other chromosome, as in figure 2,

Figure 2. Diagram of a cell in which the chromosomes may be arranged in pairs, but

the chromosomes of one pair are not exactly alike. The chromosomes of a pair may be

alike in most respects, but different in one or more features. Such a cell, in maturation,

divides into two cells which are alike in the main but differ in certain hereditary factors.

The precise hereditary composition of each cell therefore depends on how the chromosomes

are distributed to the two cells.

In such a case, in the reduction division at which only half the chromo-

somes are conveyed to each daughter cell, it is not possible to produce two

cells that are identical. Each chromosome in each of these two cells is

different from every chromosome in the other cell. Moreover, the paiy^nt is

here contributing, with respect to certain characteristics, only half of its

hereditary potentialities to any one of its offspring.

Between these extremes, in which, on the one hand, the parent hands

on all its hereditary traits to all of its offspring, and, on the other hand,

transmits only half of its possibilities with respect to certain features to

any one offspring, there are all intermediate grades. The result depends

86 A. FRANKLIN SHULL

on how the chromosomes are separated into two groups. In that cell

division in which each daughter cell receives only half the total number of

chromosomes, it appears to be a matter of chance, subject to certain

restrictions, how the half number shall be made up. If I have not made

this procedure clear, the following analogy will be useful. If it is pro-

posed to divide by chance a group of buttons, or poker chips, if that be a

more familiar figure, of a variety of colors, into two groups of six each, it is

obvious that the groups of six may be very unlike; also, that if the same

dozen buttons be divided into two groups again, the second division may

b e very unlike the first. If these buttons represent chromosomes, and

their colors stand for hereditary traits, it is clear that these traits may be

distributed in very difTerent ways to different offspring.

The chromosomes, then, because they act to some extent indepen-

dently of one another, offer an explanation of the independence of unit

characters — provided only that the things which produce these characters

are in the chromosomes. There are other reasons, equally good, perhaps

better, for believing that the hereditary factors, as they are called, are in

the chromosomes, but this additional evidence may pass.

Individuality of the Chromosomes

All this conception of the operations of heredity, in relation to the

chromosomes, was arrived at before the ten year period of which I am

eventually to speak. But certain difficulties are inherent in the concep-

tion. The number of chromosomes in the cells of an animal is strictly

limited. In man, one author fixes the number at 48, another at 24. In

other animals there is better agreement, and the number is as low as four,

or even two. In man, even the largest number suggested, 48, must be

much smaller than the number of traits which he inherits. If this be true,

the representatives of several traits must reside in the same chromosome.

The difficulty involved in this situation was that the chromosomes were

believed to he individuals. That is, the chromosomes which become dis-

tinguishable at one cell division were held to be the same identical chrom-

osomes, part for part, as were observable at the preceding cell division; and

chromosomes occurring in one individual were believed to be identical with

those of its parents. There were many facts concerning the shapes of the

chromosomes, and their behavior at various times, which lent support to

the view that they are persistent individual objects.

W.Te this regularly true, two hereditary traits represented by something

in the same chromosome would necessarily behave as a single characteristic.

They could not be independently assorted, when the chromosomes were

separated into two groups in the reduction division in the production of

germ cells, but would go together. Traits represented in different chromo-

somes would be independently assorted, but those in a single chromosome

would act as a unit.

department of summaries 87

Requirements of Proof of Linkage

Whether this condition actually existed in any animal or plant was for

a long time not known. To determine whether inherited traits were ever

bound together in groups required an animal in which differences in a con-

siderable number of characteristics existed in different individuals. It

required also a careful study of such an animal or plant to determine

whether the traits were wholly independent, or were grouped. Early in

the revival of Mendelism, an association of certain hereditary traits with

sex was demonstrated, but indications of an association of hereditary

traits with one another were long delayed. The number of traits whose

inheritance was understood, in any one species, was too small.

Then came the year 1910. In that year a fly was born — or hatched.

It belonged to the small brownish gray species which is seen every summer

day hovering about fruit stands or garbage pails. This species had

been bred for years in a number of laboratories, notably those of Columbia

University by Professor T. H. Morgan and his students. Then one

afternoon, in a bottle, appeared the fly of which I speak, which differed

from all others in the bottle, and from all of its ancestors for many genera-

tions, in having white eyes. Flies of this species regularly have red eyes.

Since 1910, other eye colors have appeared, vermilion, cherry, eosin, buff,

tinged, blood and purple being the names applied to some of them. Other

flies were produced which had unusual wings — short, blunt, crumpled, or

missing, curled up, curved down like an inverted bowl of a spoon, or spread

at an angle. Other parts of the body likewise presented variations. The

spines became forked, or reduced in number. Extra legs were produced.

The color of the body became yellow or black in certain individuals.

Physiological changes not producing any observable structural diff'erences

have also been detected. All told, over two hundred such modifications

have been discovered in this one species of fly since 1910. Most of these

characteristics were found to be definitely inherited. Fortunately, many

of these altered flies were quite healthy, were easily reared, and have been

carefully studied. The fruitfly was obviously the organism by which the

individuality of the chromosomes and their relation to heredity could be tested.

Early Demonstrations of Linkage

This test came gradually. It was found that these characteristics

were not wholly independent of one another. Thus, white eye and yellow

body-color were very closely associated with one another. When a

white-eyed and yellow-bodied fly was crossed with a normal red-eyed and

gray-bodied fly, their offspring in certain subsequent generations, in

certain cases should have shown all combinations of the two eye colors

and the two body colors with equal frequency. That is, if the four traits

were inherited independently of one another, white eye and gray body

88 A. FRANKLIN SHULL ^

should have been combined in one individual as often as white eye and

yellow body. But they were not; white eyes and gray bodies were found

together in only about one-fiftieth as many cases as would have been

expected. White eye was nearly always associated with yellow body in

these crosses.

It was discovered, also, that white eye color was associated in the same

way, though less closely, with sable body color, with club shaped wings,

and a number of other characteristics. Moreover, if white eye color was

thus bound up with a certain characteristic, yellow body color was also

associated with the same characteristic. And all characteristics that were

thus associated with white eye and yellow body were found to be linked —

that is the word Morgan uses — with one another. All these traits behave,

to some extent, as a unit. They are not absolutely bound together, but they

hang together more frequently than the chance assortment of chromosomes, or

colored buttons, or poker chips, ivould lead one to expect.

Independent Linkage Groups

Approximately forty of the more than two hundred new character-

istics that have arisen in this fly in the past ten years may safely be said

to belong to the group that is linked with white eye color and yellow body.

Long before all of these had been discovered — indeed, when only a few of

them were known — certain other new traits had come into existence which

were definitely not linked with white eye. One of these was a short

crumpled wing which has been called vestigial. In crosses which involve ves-

tigial wing and white eye at the same time, the occurrence of vestigial wing in

the individuals of subsequent generations bears no relation to the occur-

rence of white eyes in the same individuals. The chances are even, in such

crosses, that a fly with a vestigial wing will have white eyes in as large a

proportion of cases as will a fly with normal wings. Likewise, there is no

relation between vestigial wings and yellow body color. Nor is there

any association between vestigial wings and any other characteristic in

the entire group that is linked with white eyes and yellow body. Clearly,

vestigial wing is not a member of that group.

Another character that is independent of white eye color is black body

color. In crosses which should test any such su]iposed relation, the

distribution of black color of the body among the individuals is wholly

unrelated to the white color of the eye. Black body occurs with equal

relative frequency in, individuals with red eyes and white eyes. Black

body color is also independent of any other characters of the group linked

with white eye color. But black body color is associated with vestigial

wing. If a cross is made involving an individual with both vestigial

wing and black body, then in generations jiroducod by a]ipro]-)riatc crosses

among the descendants, black body and vestigial wing will occur together

DEPARTMENT OF SUMMARIES 89

much oftener than apart. That is, flies having both black body and

vestigial wing will be relatively much more numerous than flies havin^

black body and normal long wing; and much more numerous than flies with

vestigial wing and normal gray body. Vestigial wing and black body color

are clearly linked with one another.

With these two traits are also linked a number of others that concern

the wings, the body, the eyes, etc. All characteristics linked with black

body or with vestigial wing are, when tested in appropriate crosses, found

to be linked with one another. They form a distinct second group,

every member of which is linked to some extent with every other member.

This group contains about as many characters as does the first group

linked with white eyes. It must be made entirely clear that, while all the

traits of this second group are linked with one another, none of them is in

any way linked with any character of the first group to which white eye

and yellow body belong.

There is still a third group of characters, and a fourth group quite small

in numbers, which are made up, as are the first two, of characters that tend

to hang together, once they start together. All members of the third group

hang together more than mere chance would permit; and all members

of the fourth group are in like manner associated with one another more

frequently than can be attributed to accident. But no trait of the third

group is in any way bound with any trait of the first, second or fourth

groups. And no trait of the fourth group is linked to any extent with any

member of any of the first three groups. Three of these groups are rather

large, that is, include numerous characters, one is quite small.

Chromosomes and the Linkage Groups

You will have guessed long since that the reason assigned for the

linkage of these various traits is that the hereditary factors responsible

for them are located in the same chromosomes. All characteristics of the

first group are produced by something in the same chromosome. All

characteristics of the second group are likewise represented by something

in one chromosome. But that chromosome is a different one from the

chromosome that produces the characteristics of the first group. Each of

these groups owes its existence as a group to one chromosome, which is a

different chromosome for each group.

In this connection you will care to know something about the chromo-

somes of this fly. Fortunately they are well known. Each cell has eight

of them (figure 3), but when, in the formation of germ cells, the reduction

division divides these into two groups, there are only four in each germ

cell. Three of these chromosomes are large, and one quite small, and three of

the linkage groups of characters are large, and one small.

Assuming that the hereditary factors for one group are all in one

chromosome, and that that is the cause of their linkage with one another,

90

A. FRANKLIN SHULL

Figure 3. The chromosomes of the female fruitfly Drosophila melanogaskr. In the

body cells and immature germ cells there are eight chromosomes, two each of four kinds.

In maturation, at the reduction division, the number is reduced to four, one of each of the

four kinds. Three of the chromosomes are large, and one is small.

what becomes of the idea of individuality of the chromosomes? It must

be modified, of course. As previously pointed out, the characteristics of

each group are not absolutely bound together, they merely occur together

more frequently than chance would permit in the case of independent

characteristics. That is, once they are transmitted from parent to off-

spring in conjunction with one another, they separate from one another

thereafter less frequently than would be expected. But if their factors

are in the same chromosome, how can they separate at all?

Breaking the Linkage Grouts

To make the proposed answer to this question clear it must be stated

that when this separation does occur, there is a fairly even exchange.

That is, when white eye is separated from yellow body with which it had

been associated, some other eye color takes its place and is thereafter as

closely linked with yellow body color as was the white eye color before.

This is always the case. Whenever a trait is removed from assoeiatiou with

another trait, its place is taken by a trait related to the same part of the body.

Eye color is exchanged for eye color; one form of wing is replaced by an-

other form of wing.

This exchange is made possible, presumably, because of the approximate

duplication of the chromosomes in each cell. It has already been pointed

out that the chromosomes of a cell may be such that they are exactly

alike, two by two (figure 1). But even where the chromosomes arc all

DEPARTMENT OF SUMMARIES 91

diflferent from one another (figure 2), nevertheless they can be arranged in

pairs of twins such that the members of one pair differ from one another

in only one or a few features, but are alike in a host of others. One of them

may, for example, include a representative of vestigial wing, the other of the

normal long wing, but be alike in everything else. Or they may differ with

respect to color of body and color of eye, and be alike in all other respects.

The two chromosomes have to do with the same parts of the body, and no

other chromosomes of the cell are concerned with those traits in the same

way. The chromosomes are truly capable of arrangement in pairs of twins.

Mechanism of Crossing-over

This arrangement in pairs is not purely a figurative one, it is at certain

times an actual bodily one. At a certain time in the formation of the

germ cells, these chromosomes come together side by side. What they

look like in this operation, is known in relatively few forms. In one of these

the chromosomes are long slender threads, and the two twins twist about

one another in loose spirals (figure 4). This is not an isolated case. It is

iSS^P^^

Figure 4. Some of the chromosomes of Batracoseps twisting about one another in

spiral form prior to the reduction division. The chromosomes thus twisting together contain

factors for the same hereditary characters. Whether they untwist in the reduction division,

or separate in some other fashion, is not known from observation. {M odified from Janssens.)

not impossible that, in many or most animals, the twin chromosomes twist

about one another at this stage of development. Later they separate

from one another in some fashion at what we have called the reduction

division. How this separation takes place is not known from direct

observation, but several possibilities exist. The chromosomes may

unwrap completely and be the same chromosomes as before they twisted.

Or they may adhere at points, and the two sides of the spiral in that region

exchange places. This is a very important conception, put forward by

Professor Morgan and his students, but before it can be developed certain

other considerations must be presented.

92

A. FRANKLIN SHULL

The hereditary factors contained in a chromosome are, Professor

Morgan believes, arranged in linear order, like beads on a string (figure 5).

Every cell in an animal's body has a chromosome in which these "beads"

are the same as those of one chromosome in each of the other cells of the

Vellou/ Bodt/

Nhite £ye

I Graif Body

Red E(/e.

No/^mall^inq

yNorwall^inq

Venmili on

£-ye

Mmia/ure

k/ino

Red Eye

I Mmiafurc

Rudirner\-

Forked

Br/sf/es

Comp/e^eEye

Rijdimen-

fory U/ina

Forked

Bristles

Bar Eye

Figure 5. Diagram of a pair of chromosomes of the female fruilfly Drosophila melano-

gasler. The hereditary factors are arranged in a single row in each chromosome. Both

chromosomes have hereditary factors for the same characters. The factors for one character

are placed at the same level in both chromosomes, so that when the chromosomes meet in a

pair the two homologous genes are side by side. Not all the known factors are represented.

{From Principles of Animal Biology, by Shiill, La Rue and Ruthven. McGraw-Hill Book Co.)

body. In the same cell with it is another chromosome in which the

hereditary factors are precisely the same, or at least they concern the same

parts of the body. The hereditary factors in these two chromosomes are

held to be arranged in the same order, and to lie at the same level. So that,

if the chromosomes are placed side by side, or twisted about one another,

the two hereditary factors Jor the same part of the body are side by side.

If two such chromosomes twist about one another, as has been de-

scribed, and then in separating are not unwrapped carefully, they may

exchange hereditary factors. If in one of these chromosomes were a factor

for white eye and one for sable body color (figure 6), some distance apart so

that the breaking point occurred between them, the linkage that formerly

existed between these two characteristics would be broken. Where they

DEPARTMENT OF SUMMARIES

S

93

Figure 6. Crossing-over between two chromosomes containing factors for eye color and

body color. One chromosome has factors for white eye (w) and sable body {s); the other

has factors for red eye (W) and gray body (S). If these chromosomes adhere and break at

some point between the pairs of factors, after the reduction division one chromosome con-

tains factors for white eye (w) and gray body (S), the other has factors for red eye (W)

and sable body (s).

had formerly necessarily passed to the same individual, they would now

necessarily pass to different individuals. There is also much to prove that

the chromosomes may break twice, or at three places instead of only one.

If one of the two chromosomes that twist about one another (figure 7) has

MB

Figure 7. Double crossing-over in a pair of chromosomes. The chromosomes adhere

and break at two points in their length, so that after the reduction division each chromosome

is made up of three fragments, two from one of the original chromosomes, one from the

other. In the original chromosomes the factors w, m and B were linked, as were also W,

M and h. After the crossing-over, w, M and B form one linked group, IF, m and b the other.

See text for explanation of the s>'mbols.

in it factors for white eye {w), miniature wing {m), and bar eye {B), the

other the contrasted normal characteristics which are red eye {W),

94 A. FRANKLIN SHULL

long wing (-M), and round eye (b), and these chromosomes break at two

points as they separate from one another, two of the three linked characters

may be still linked together, but the third separated from them. The

separation of a here4itary factor from another with which it was linked is

called crossing-over.

Whether a pair of twisted chromosomes shall break at one point or

another is, with certain restrictions, held to be a matter of chance. If

they break between the factors for black body and vestigial wing, those

factors will be released from linkage with one another; that is, crossing-over

between these two characteristics occurs. These chromosomes may

break at any number of places not between the factors for black body and

vestigial wing, and the two traits will remain linked as before.

Mapping Chromosomes

Inasmuch as breakage presumably occurs at different places in hap-

hazard fashion, crossing-over between two traits is likely to occur often if their

factors are far apart on the chromosome. Conversely, if the factors are very

near together they are seldom separated. It is very easy, by making appro-

priate crosses, to pick out immediately those individuals in which certain

characteristics, usually associated, have been separated, and hence in

whose chromosomes breaking has occurred in a given region. By counting

these individuals one may ascertain whether crossing-over between vermil-

ion eye and club wing, for example, is frequent or rare; and can judge,

therefore, whether the factors for these characteristics are far apart or near

one another. By means of such experiments, it has been shown that white

eye and yellow body seldom separate; they do so in only one out of a

hundred chances, whereas they should cross over fifty times out of a

hundred if they were independent of one another. Black body and

vestigial wing, on the contrary, separate much more frequently, that is.

about seventeen times out of a possible hundred. White eye and yellow

body must therefore be very close together, black body and vestigial wing

must be rather far apart. On the basis of such computations entire

chromosome maps have been prepared. Such maps have been in existence

for years, having been gradually developed, and altered as new evidence

is procured. An abridged map of one of the chromosomes of the fruitfly

is given in figure 8.

As new characters appear in this species, experiments are performed to

determine in which chromosome their factors are, by determining with which

other characters they are linked. And when that chromosome has been

discovered, the place in it occupied by the new factor is next to be found.

In locating the new factor, it may be necessary to alter the supposed place

of certain other factors. That has happened lime and again, for at first the

location can be only tentative.

DEPARTMENT OF SUMMARIES 95

0

Wiitt eye

No croi-ii/om

Vermilioneyc

tiiiya

^obl« bod

6oir CyG

VcllOvv body

BfJd vcr,

Cut ^,n.^

T<xr\ b&dy

Garnet g yc:

Forked bnsfic;

Figure 8. Abridged map of one of the chromosomes of the fruitfly Drosophila melanogas-

ter, showing approximately the distances between the factors. The distances are determined

by the number of times crossing-over occurs between the hereditary factors. Attention is

especially called in the te.xt to the distance between the factors for vermilion e^-e and sable

body, and that between garnet eye and forked bristles.

Linear Order of Factors

While crossing-over can be easity demonstrated, it is admittedly pure

hypothesis that it is accomplished by twisting of the chromosomes. It is

also pure hypothesis that the factors are arranged in linear order, and that

the amount of crossing-over between any two depends on the distance

between them. However, any hypothesis is valuable in proportion to

the number of things it explains, and a hypothesis that explains many

things and is contradicted by nothing, has traveled a long way toward

proof. Judged by these criteria, let us examine the situation of the

hypothesis of the linear order of the hereditary factors and the twisting of

the chromosomes.

First, assume that the amount of crossing-over between two characters,

A and B, has been determined, and that from this amount the supposed dis-

tance between the factors for A and B, in terms of some arbitrary unit of

measurement, has been computed. Suppose also that the distance be-

tween B and a third factor C has been similarly determined. If these

determinations are actually computations of distances, then these two

calculations fix the distance also between A and C, and it should be possible

96 A. FRANKLIN SHULL

to predict how great a proportion of crossing-over should occur between A

and C, before the experiments to test it have been performed.

These predictions, when applied to short distances, have proven

remarkably accurate. For example, as has been stated, in the first

chromosome, breaking of the chromosome between the factors for yellow

body and white eye occurs in about one per cent of possible cases. Cross-

ing-over between white eye and bifid wing vein has been computed to occur

in about five per cent of possible cases. If the map of this chromosome

has been properly constructed (figure 8), crossing-over between yellow

body and bifid wing vein should occur in about six per cent of cases.

Within a small fraction of a per cent, this prediction is verified by experi-

ments directly testing it. Similar verification has been obtained in

numerous other groups of factors, and in no case where only small dis-

tances are involved have there been any serious discrepancies.

Errors in Long Distances

The fact that only in cases where small distances are computed is the

correspondence between prediction and fact very close may at first seem

to weaken the evidence in favor of the theory. On the contrary, lack of

close correspondence between prediction and discovery ' where long distances

are concerned, is an important part of the confirmation of the theory. I have

already pointed out that the same two chromosomes, when they meet,

may break at several points. Suppose that, in this case, the two chromo-

somes differed from one another in only two factors, white eye and bar eye

(see figure 7, w and B), and that there were no identifying factor between

them. If, in these premises, crossing-over should occur at two points be-

tween the factors for white eye and bar eye, the two factors would never-

theless be in the same chromosome after the chromosomes separated, and

the experiment would not reveal the fact that crossing over had occurred

at all. Crossing-over might occur four times, or six times, or any even

number of times, and the two factors would still be left in the same chrom-

mosome. When the two factors are separated by long distances, multiple

crossing-over between them is likely to occur, and only a part of it is

delected. Such factors would appear, by the usual computations, to be

much nearer one another than they actually are. When a new factor is

discovered, its location may be tentatively determined with reference to

some other well known factor. If it turns out to be rather far from the

factor first chosen as a zero point, the test may be repeated with another

known factor which is probably near it. It has invariably happened that

these later computations increase the distance as determined from the

first long-distance test. That is simply because, in the second computa-

tion which concerns only a short distance, the investigator is discovering all

the cases of crossing over, instead of only a fraction of them, and the dis-

DEPARTMENT OF SUMMARIES 97

tance is necessarily made to appear greater. So regularly has it happened

that first computations of long distances have had to be increased as

intermediate short distances are determined that the investigator is now

able to guess with considerable accuracy how much a long distance diverges

from the truth, and to fix with a good deal of precision the probable loca-

tion of a new factor before the second computation is made. Lack of

precision in the calculation of long distances is not, therefore, a weak point

in the argument. Indeed, there would be something the matter with the

theory if long distances could be determined as accurately as short ones.

Points of Cross-Over Never Near Together

The theory of linear arrangement of the factors, and of the twisting of

the chromosomes, carries with it certain corollaries. It is scarcely pos-

sible that crossing over at one point of a chromosome is entirely inde-

pendent of crossing over at other points in the same chromosome. Thus,

if two chromosomes break at two points, a chunk is removed from the

middle of each chromosome and transferred to the other. The size of this

chunk is probably not entirely free of limitations. It extends from one

point of crossing over to the other. How near or how far apart these

breaks can be may depend on how tightly the chromosomes are twisted, or

at how frequent intervals they adhere intimately to one another. No

upper limit is set to the distance between cross-overs, but a lower limit

is certainly to be expected. The size of a piece of chromosome that

can be removed, by this method, to another chromosome presumably

can not fall below a certain minimum. That means that if breaking

occurs at one point, it is not likely to occur at a nearby point at the same time.

Fortunately, owing to the large number of factors now known in

some of the chromosomes, this interference of crossing over at one point

with crossing over at a nearby point can be tested. Thus, in one of

the chromosomes (figure 8) the factors for vermilion eye and sable body

color are at such a distance from one another that crossing over between

them occurs once in ten times — in ten percent of cases. In a nearby

region of this chromosome are two other factors, one for garnet eye,

the other for forked bristles, so placed that crossing over occurs between

them in twelve per cent of possible cases. If the breaking of the chrom-

osome in one point has no bearing upon its breaking at another point, any

chromosome that is severed between vermilion and sable should have

twelve chances in a hundred of breaking also between garnet and forked.

But experiments on a large scale show that, if crossing over occurs in the

vermilion-sable region, it occurs also in the garnet-forked region in only a

little over one case (1.2 to be more precise) out of a hundred. In like

manner, if the various breaks of the chromosome are wholly independent

of one another, out of a hundred chromosomes that have broken between

98 A. FRANKLIN SHULL

garnet and forked, ten should also be found to have broken between

vermilion and sable. As a matter of fact, only one of the hundred breaks

in the latter region. Crossing over in one of these regions obviously inter-

feres with crossing over in the other region.

These two regions are near one another. The reason for the inter-

ference may be attributed, whether correctly or not, to the closeness or

looseness of the spiral winding of the chromosomes about each other.

When the two chromosomes adhere at one po'nt, unless they are very

tightly wound, they are not likely to adhere at another point except at

some distance. If this explanation is the correct one, interference of cross-

ing over should be most marked when the two regions are near one another,

less marked when the regions are farther apart. That has indeed proven

to be the result. Regions farther apart than these have been tested for

simultaneous crossing over, and the degree of interference has decreased

as the distance between the regions tested increased, up to a certain limit.

This limit was reached when the two regions studied were far enough

apart to account for about 46 per cent of crossing over between them.

When regions farther apart than this were studied, then interference

became greater again.

Variation in Frequency of Crossing-over

The idea that the frequency of crossing-over within a certain region

depends upon the tightness with which the chromosomes twist has found

expression in the explanation of another phenomenon. When this fre-

quency of crossing over is determined by experiments, the results usually

have a considerable degree of uniformity. The amount of crossing

between vermilion and sable, for example, has never diverged very much,

in an experiment involving large numbers of individuals, from ten per

cent. However, certain strains of these flies have been found in which

the ratio of crossing over differs very considerably from that found in

other stocks. It is fairly uniform within the aberrant slock, but is differ-

ent from other strains. This capacity for, let us say, a smaller amount of

crossing over is, furthermore, transmitted to the offspring as a permanent

family character. One inter])rctation put upon this phenomenon has been

that some inherited feature, doubtless of a physiological nature, causes

the chromosomes, when they meet in pairs, to wrap more loosely, about

one another; or perhaps Tnerely to adhere less frcciucntly.

Further elaborations of this hypothesis of linear order of the hereditary

factors, and of the twisting of the chromosomes, would be available if

desired. Perhaps, however, the pur]H)se which this lecture is designed to

serve has been fulfdled. If it has shown that the study of heredity has under-

gone a very considerable change in the past decade, it has accomplished one

of its aims. If instead of com])osing you to slumber it has convinced you

DEPARTMENT OF SUMMARIES 99

that the modern study of heredity is no longer a subject with which to

lull oneself into an afternoon nap, it has attained another of its objects.

I have not conducted you quite to the limits of present knowledge. Cer-

tainly we have not been anywhere near the confines of speculation. The

discussion should have shown that in recent years the complexity of fact

and theory in heredity has enormously increased. The progress made in

this period is, I believe, unquestionably proportionately greater than has

been made in the same period in either physics or chemistry. Biology, in at

least this one division of it, has taken a long stride toward destroying

the significance of that relative term by which the sciences of physics,

chemistry, astronomy, etc., are so often fondly designated by their follow-

ers, namely, the "exact sciences." So complex now are the known phenom-

ena of heredity, and yet in such close agreement are the multitude of

facts of experiment and observation with the chain of hypotheses devel-

oped to explain them, that one who would contemplate the harmony

of the universe may now reasonably strain his ear to catch, not the music

of the spheres, but the concert of the chromosomes.

Bibliography

The following publications include some of the original work along each of the principal

lines of development referred to in this article. Most of them contain references to other

papers, so that fuller bibliographies may be prepared by those who require them.

Bridges, C. B.

1913 Non-disjunction of the sex chromosomes of Drosophila. Jour. Exp. Zool.,

15:587-606.

1916 Non-disjunction as proof of the chromosome theory of heredity. Genetics, 1:1-52,

107-163.

1917 An intrinsic difficulty for the variable force hypothesis of crossing over. Amer.

Nat., 51:370-373.

1917 Deficiency. Genetics, 2:445-465.

1919 Vermilion-deficiency. Jour. Gen. Physiol, 1:645-656.

Bridges, C. B., and T. H. Morgan.

1919 Contributions to the genetics of Drosophila melanogaster. II. The second

chromosome group of mutant characters. Carnegie Inst, of Wash., Pub. 278.

GOLDSCHinOT, R.

1917 Crossing over ohne Chiasmatypie. Genetics, 2:82-95.

HoGE, M. A.

1915 Another gene in the fourth chromosome of Drosophila. .\mer. Nat., 49:47-49.

Metz, C. W.

1914 Chromosome studies in the Diptera. I. A preliminary survey of five different

types of chromosome groups in the genus Drosophila. Jour. Exp. Zool.,

17:45-59.

1916 Chromosome studies in the Diptera. II. The paired association of chromosomes

in the Diptera, and its significance. Jour. Exp. Zool., 21:213-279.

1916 Chromosome sttidies on the Diptera. III. Additional types of chromosome

groups in the Drosophilidae. Amer. Nat., 50:587-599.

100 A. FRANKLIN SHULL

Morgan, T. H.

1912 The linkage of two factors in Drosophila that are not sex-linked. Biol. Bull.,

23:174-182.

1914 No crossing over in the male of Drosophila of genes in the second and third pairs

of chromosomes. Biol. Bull., 26:195-204.

1915 Localization of the hereditary material in the germ cells. Proc. Nat. Acad. Sci.,

1 :420-429.

1915 The constitution of the herditary material. Proc. Amer. Phil. Soc, 54:143-153.

Morgan, T. H., and C. B. Bridges.

1916 Sex-linked inheritance in Drosophila. Carnegie Inst, of Wash., Pub. 237.

MULLER, H. J.

1914 A gene for the fourth chromosome of Drosophila. Jour. Exp. Zool., 17:325-336.

1916 The mechanism of crossing-over. Amer. Nat., 50:193-221, 284-305, 350-366,

421-434.

Plough, H. H.

1917 The effect of temperature on crossing-over in Drosophila. Jour. Exp. Zool.,

24:147-209.

Sturtevant, A. H.

1913 A third group of linked genes in Drosophila ampelophila. Science, N. S., 37:990-

992.

1913 The linear arrangement of six sex-linked factors in Drosophila, as shown by their

mode of association. Jour. Exp. Zool., 14:43-59.

1915 The behavior of the chromosomes as stucHed through linkage. Zeit. f. induk.

Abst. Vererb., 13:234-287.

1919 Contributions to the genetics of Drosophila melanogaster. III. Inherited

linkage variations in the second chromosome. Carnegie Inst, of Wash., Pub.

278.

Weinstein, a.

1918 Coincidence of crossing-over in Drosophila melanogaster (ampelophila). Genetics,

3:135-172.

DEPARTMENT OF METHODS, REVIEWS, ABSTRACTS,

AND BRIEFER ARTICLES

A NEW MICRO-SLIP

It is hardly necessary to add anything to the illustration of this con-

venient, strong and economical micro-slip, as it almost explains itself.

The body of the slip is made of sheet tin cut into parallelograms

three inches long by one and one-quarter inches wide. A circle about

thirteen-sixteenths in diameter is then punched out of the exact center

and one-eighth inch on each long side is bent over forming two flanges into

which the three finished units are slipped.

Object is mounted on a 25 mm square cover-glass of medium thickness

in the regular manner. As the 25 mm squares are not very strong when

handled by themselves, it is better to fasten them to a square piece of

clear glass, before mounting, by squeezing down while wet, and then

drying the upper side. This will give the necessary strength and support

and they, will be held firmly by capillary attraction.

When the mounting is completed, the 25 mm square is slipped into

place at center of tin holder and a square piece of bristol board or card

board slid into each end. The corners of the flanges are then squeezed

tight with pincers and slide labeled.

A drop of transparent cement drawn under the flanges by capillary

attraction, after the mount is completed, holds everything firmly in

position and prevents any slight future movement of the units.

These slips can be easily made by any tin-smith.

The finished product is much stronger than the ordinary glass slip

and the mount will seldom break on falling to the floor.

A ventral view of the mounted object can be had, just as easily as the

dorsal view, by simply reversing the slide.

I am indebted to my friend Mr. H. K. Harring of Washington, D. C.

for this slip originally, and have been using them for some years with

every success.

101

102 FRANK J. MYERS

The collection of Rotifera, about 500 species, in the American Museum

of Natural History, New York City, are mostly mounted on these slips

and have proven successful in all particulars.

Frank J. Myers.

Research Associate in Rotifera,

American Museum of Natural History

Since the above was written I learn the slip described is the invention of Dr. N. A.

Cobb, Washington, D. C. It may be purchased from the Spencer Lens Co., Buffalo, N. Y.

I

KILLING, STAINING AND MOUNTING PARASITIC

NEMATODES*

By

H. G. May

In some work on Heterakis papulosa of the domestic fowl I found that

the ease with which fluids will penetrate the cuticula depends much on

the method by which the worms have been killed.

Parasitologists are mostly following Looss in killing nematodes by

means of hot solutions, usually of alcohol. This has the advantage of

straightening the worms out as they die. The straightening seems to be

due to the heat, as hot water will have the same effect. Cold concentrated

killing solutions will usually cause the worms to die in very much contorted

shapes.

When nematodes are killed by either of the above methods staining is

rendered very difficult and mounting in balsam usually is impossible unless

the cuticula is broken in one or more places. The final mounts are usually

of a very inferior nature even tho all possible precautions are taken in

changing from one fluid to another.

This resistance to penetration is not a normal function of the nematode

cuticula as in the living worm fluids seem to penetrate the cuticula very

readily. The resistance is due to a change in the character of the cuticula.

This change is also indicated by the curling up or rolling up of free parts of

cuticula such as wings or a pointed tail. These parts behave as tho the

cuticula becomes semifluid during the killing process and then sets in an

impervious condition.

This change can be prevented to a great extent by employing the

method used by Dr. Cobb in killing free living nematodes. The essential

point seems to be to start with a very weak solution. The nature of the

chemical substance used is undoubtedly also of great importance. I have

obtained very good results only with ethyl alcohol. Methyl alcohol and a

solution of mercuric chloride both gave very inferior results; but this may

have been due to the use of too strong solutions. Not only does this

method prevent the change in the cuticula, but it also usually gives just as

straight specimens as those killed in hot solutions.

In my work I still find it more convenient to use the string siphon

differentiator as first described by Magath (Trans. Amer. Micr. Soc,

35: 245-256) and with the modifications described by me (111. Biol. Mon-

*Contribution No. 289 of the Agricultural Experiment Station of the Rhode Island State

College.

103

104 H. G. MAY

ogr., Vol. 5, No. 2, p. 13) than to use Dr. Cobb's apparatus. An attempt

to use his differentiator almost invariably results in the loss of valuable

material because it cannot be found after the process has been com-

pleted. If the material is abundant so that the loss of a few specimens is

of no consequence his method has the advantage of using less fluid and

occupying less space, I find difficulty also in regulating the flow of liquid in

his differentiator as that depends not only on the size of the capillary outlet

but also on the weight of the column of liquid and the concentration of

the alcohol, and the capillary tube may become clogged at any time.

In the string siphon method the specimens may be placed in a dish

the bottom of which can be examined everywhere by means of a magnify-

ing glass, binoculars or even a very low power of a compound microscope.

The specimens need not be disturbed during the entire process of dehy-

dration and clearing. By the method I am now using a single nematode

of microscopic dimensions may be placed in a deep embryological watch

glass while alive, and killed, stained, dehydrated, cleared and infiltrated

with balsam without being disturbed. The only transfer necessary

is that from the watch glass to the final mount on the slide.

By this method the specimens are placed in a watch glass or small

stender dish in eight to nine tenths per cent salt solution. By means of a

string siphon an amount of 5-7 per cent ethyl alcohol equal to four or five

times the volume of salt solution is passed over the specimens. This is

followed by an equal amount of 10 or 15 per cent alcohol and so on. Each

time the concentration of the alcohol is increased by 5 to 10 per cent. The

size of the string should be so regulated that it requires from 6 to 12 hours

for one change of fluid to take place.

The staining may be done anywhere in the course of the dehydration.

I usually stain with Delafield's or Boehmer's hematoxylin in 70 per cent

alcohol with the addition of a small amount of acetic acid to aid penetra-

tion and to avoid the necessity of destaining. This is followed by a weak

solution of sodium or potassium acetate in 75 or 80 per cent alcohol. The

dehydration is then continued.

Absolute alcohol is followed by 1-3 xylene in alcohol, 2-3 xylene in

alcohol and pure xylene. If specimens are to be sectioned they are now

imbedded in paraffin by the method described in my paper quoted above.

If they are to be studied as cleared specimens or mounted in balsam they

are next passed from xylene to synthetic oil of wintergreen just as they were

passed from alcohol to xylene. This gives the specimens a maximum

amount of clearing and makes excellent preparations for study at that

time. Mounting in balsam renders the specimens more opaque and makes

them appear more like they did in xylene.

To mount in balsam it is usually best to infiltrate the specimens

first by removing all but a thin layer of the oil and then adding a small

DEPARTMENT OF METHODS 105

lump of dry Canada balsam every day or so until the solution has acquired

about the consistency of the balsam to be used for the mount. For

permanent mounts it is best to use balsam dissolved in xylene even tho

this does not retain the transparency of the object because balsam in oil

of wintergreen requires months to harden. Gum dammar may be used

for mounting but not for the process of infiltration as it does not dissolve

completely in the methyl salicylate.

A NEW LOCALITY FOR S POX GILL A WAGNERI POTTS

The description of this species in 1889 was based on material collected

in a creek on the southwestern coast of Florida. The sponges were

found encrusting barnacles and tubes of Serpulae. The presence of these

marine forms in a fresh-water creek was assumed to be due to occasional

backing up of salt water from the Gulf under the influence of strong

southwest winds. I have found no reference to the occurrence of the

species elsewhere in North America, but it has been recorded as occurring

in Brazil.

Fresh-water sponge material recently received from the Southern

Biological Supply Co. and collected in Lake Pontchartrain, near New

Orleans, includes specimens of S. wagneri, and again the colonies are

associated with barnacles. P. Viosca Jr. has informed me that under

certain conditions salt water gains entrance, and varying degrees of

salinity occur at times in different parts of the lake. It seems probable

that representatives of the species may be found in suitable localities in

other states along the Gulf Coast.

Frank Smith.

University of Illinois.

106

SOME INTERESTING STUDIES ON SPIDER ANATOMY

By

E. W. Roberts

On studying the spider Anglena I was struck by the lack of conformity

of the material with descriptions of the class given in prominent text

books. For instance, in Parker and Haswell's Zoology, vol. 1, page 615,

the pedipalpi of spiders is described thus: "The pedipalpi (Fig. 506, B) are

elongated, and end in simple extremities; in the male (Fig. 507) the terminal

joint is modified to serve for the reception and transference of the sperms."

In the pedipalpi of the male Anglena, we find the third from the end, not

the terminal joint, serving as a very highly modified intromittent organ.

The pedipalpi (Fig. 1) are composed of at least 9 separate segments as I

have numbered them from the extremities inward. Segment 1 is modi-

fied into a terminal clasper, or claw. Segment 2 is a much enlarged gland

and olfactory joint; there are two large areas of perforated openings, one

of which gives off sexual odors, while the smaller perforated area consists of

olfactory cells. This joint during sexual contact is placed directly over the

stigmati or air chambers which open into the book-lungs. Thus in this

case the book lungs function in an entirely new light as olfactory organs.

Segment 3 is modified into a very complicated intromittent organ, which

is shown greatly enlarged in photo No. 2.

In preparation for mating, the male spins a mat of web which is held

under the sexual vents and the sperms emitted thereon. This web with

its contents is then brought forward and the end of the joint marked A is

inserted in the fluid mass which is sucked through slit apertures into the

large coiled storage glands on which I have indicated the direction of the

flow by arrows. The large gland makes about one complete turn and

ends in a minute tube which terminates outward in the true intromittent

organ marked B. During copulation the male, watching his chance, rushes

behind the female and grabs her around the neck of the abdomen and

inserts the terminal B in the small vaginal vents which are shown as a

pair in photo No. 3 and marked A-A.

The stigmati c-c are situated external to the vaginas and are marked

B-B. In this position the large gland organs of joint 2 come into position

over the stigmati of the further side, while joint 1 clasps its claw further up

the side.

Segments 4-8 are well represented in the illustration. Segment 9 is

enlarged (Fig. 4) forming two lips which contain the most highly developed

nervous system of any organ in the body.

107

108 E- ^V. ROBERTS

The nerve cord spreads out fan-wise, divides into thousands of termin-

als which end in the minute cuticular hairs or spines. These hairs dip

into the blood of the victims and are probably gustatory in function.

In Parker and Haswell's Zoology, vol. 1, page 612, order 6, Araneida.

"Arachnida in which the body is composed of an undivided cephalothorax

and an unsegmented abdomen." The claims of most textbooks on the

"undivided head and thorax" of this class, may suit the men who pro-

posed the term of cephalathorax, but to students of anatomy it is more or

less unfounded as I will try to show. It is doubtful if the head structures

and the thorax structures have ever coalesced or fused as some casual

outer views would indicate. The neck of spiders is as large as the thorax

and is doubtless made so by the fact that in bulk with its appendages it is

as large as the true thorax. I have no doubt that the full sized neck of

spiders more nearly approaches the ancestral types of Arthropoda than

that of Diptera which are reduced in some cases to a mere stalk. But

this would not keep a spider from having as true and anatomically com-

plete head as a fly or bee. Fig. 5 is a long vertical section of the head

and thorax of a female Anglena spider. The line marked E is the true

division of the head and thorax structures, there is no sign of structural

coalescence. In the thorax, figures 1-4 indicate the four legs and the

corresponding muscles which move them. A glance will show the head

and thorax structures nearly balanced in bulk, but entirely distinct region-

ally. The pedipalpi (A) form the lower lip, the lower neck fold being

between this and the fourth thoracic leg. B is the relatively enormous

chelicerae, the eye and the poison gland of the chelicerae are represented

at B, C, and D. Fig. 6 shows the head of a male Anglena spider and several

additional features. On the side of the head prominent folds of neck skin

project forward above the eye C and is designated E. This may be the

same neck fold which bending front or back is called a carapace. A is

the pedipalp base and B the chelicerae. The brain ganglia of the head

(D) lie forward of and in close contact with the thoracic ganglia.

In spiders the nerve ganglia of the head and thorax form one long

brain body, but there is no coalescence or fusing of the ganglia. The nerve

trunks of all the head appendages still run to their proper brain lobes and

are easily followed out in sectioned material. The only thing that has

happened to the ganglia is the shortening of the trunks between ganglia and

their approachment to each other.

In Parker and Haswell's Zoology, page 612, order 7, Acarida we find

the following "Arachnida in which the body exhibits no division into

regions." And on page 616 "In the Acarida or Mites and Ticks (Figs.

508 and 509 the distinction into regions is no longer recognizable." In

my specimens of Trombidium fuliginosum there is a small but distinct

fold on the ventral side between the abdomen and thorax, while the

6^

DEPARTMENT OF METHODS 109

Stigma lie in their proper place just to the rear of the fold, the same position

as they take in spiders. In the fig. 508 of the itch mite Sarcoptes scabiei

which they give the abdominal fold and the stigma just to the rear are

plainly in view. These points are of minor importance, but, if they are

correct anatomy why would they not be correct for teaching purposes.

Explanation of Plate

Fig. 1, Anglena — male spider — organs pedipalpi; 1, terminal claw; 2, scent gland. 3, intro-

mittent organ. 4, large oval clasping segment; 5, 6, 7, 8, small segments; 9, highly devel-

oped segments which form lips for the mouth.

Fig. 2. Enlarged view of intromittent segment. A is the intake of the storage tube,

the arrows indicating the direction of the flow. B is the outlet of the storage tube which is

furnished with a shoulder to prevent too deep insertion.

Fig. 3. Section of anterior abdomen of female spider, cut through stigmas and vaginas.

A-A, vaginal openings, B-B, stigmas; C-C, booklungs; D, liver; E, heart.

Fig. 4. Section through head of male spider. A-A, lips; B-B, chelicerae; C-C, poison

glands; D, mouth.

Fig. 5. Long vertical section of head and thorax of female spider. A, lower lip. B,

chelicerae; C, eye; D, poison gland; E, neck; 1, 2, 3, 4, legs and the muscles which move

them.

Fig. 6. Long vertical section of head and thora.x of male spider. .4, lip or pedipalpi;

B, chelicerae; C, eye; D, brain; E, neck fold or carapace.

ANNUAL REPORT OF THE TREASURER OF THE

AMERICAN MICROSCOPICAL SOCIETY

December 17, 1920 to December 24, 1921

Receipts

Balance on hand December 17, 1920 $444.19

Dues received for Volume 39 or before 75 .00

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Sales of Transactions, duplicates, back numbers 121 .01

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EXPENDITUKES

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Balance on hand 592 .71

Total 1722 . 20

December 24, 1921.

Examined and found correct.

March 27, 1922.

W. F. Henderson, Treasurer.

Raymond J. Pool

ROBT. H. WOLCOTT

Auditing CommiUee.

110

CUSTODIAN'S REPORT FOR THE YEAR 1921

Spencer-Tolles Fund

Balance reported for the year $6617 .66

Contribution 1.76

Interest and dividends 450 . 14

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Total 9104.59

Increase during the year ' $2486.90

Totals

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All contributions 802 .03

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Investments

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Item "Interest and dividends" (1921) shows net receipts and accruals after payment of

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ing, respectively.

Having examined the above account, in connection with cash account covering the

same period, and having compared the receipts and expenditures shown therein, with the

vouchers and with report for previous year, we find it correct.

F. E. Ives

Edward Pennock

Auditing Committee

111

I y-^

'1^

TRANSACTIONS

OF THE

American

Microscopical Society

Organized 1878 Incorporated 1891

PUBLISHED QUARTERLY

BY THE SOCIETY

EDITED BY THE SECRETARY

PAUL S. WELCH

ANN ARBOR, MICHIGAN

VOLUME XLI

Number Three

Entered as Second-class Matter August H, 1918, at the Post-office at Menasha

Wisconsin, under Act of March 3, 1879. Acceptance for mailing at the

special rate of postage provided for in Section 1 10?, of the

Act of October 1, 1917, authorized Oct. 21, 1918

iSife dallrgiate Prraa

George Banta Pdblishing Company

Menasha, Wisconsin

1922

TABLE OF CONTENTS

For Volume XLI, Number 3, July, 1922

Notes on the Excretory System in Aspidogaster conchicola, with two plates, by E. C.

Faust 113

A New Cestode from Liparis liparis, with one plate, by Edwin Linton 118

A List of the New Gregarines described from 191 1 to 1920, by Minnie Watson Kamm ... 122

Department of Methods, Reviews, Abstracts, and Briefer Articles

Abnormal Earthworm Specimens, Ildodrilus subrubicundus and H. tenuis, by

Frank Smith 153

Substitutes for Absolute Ethyl Alcohol, by L. E. Griffin 155

/ 1

TRANSACTIONS

OF

American Microscopical Society

(Published in Quarterly Instalments)

Vol. XLI

JULY, 1922

No. 3

NOTES ON THE EXCRETORY SYSTEM IN ASPIDOGASTER

CONCH ICO LA""

By

Ernest Carroll Faust

Peking, China

Aspidogaster conchicola is a cosmopolitan parasite. It is commonly

found in Anodon in Central Europe and is the most common parasite of the

Unionidae in North America. It has at times been described from gastero-

pods and infrequently from fishes. Examinations from several distinct

centers in China reveals the presence of the fluke in the following host

species:

In the case of the lamellibranch the worms were found parasitic on the

renal organ; in the gasteropods they were dissected out of the lymph

sinuses of the liver or taken from the spermary. In both the fish and the

turtle the worms occurred in the intestine.

The finding of Asipidogaster conchicola in several regions in China adds

new records of distribution for the species, while the presence of the parasite

in the intestine of Amyda sinensis allows one to record a new host.

*Contribution from the Parasitolog)' Laboratoty, Department of Pathology, Peking

Union Medical College.

113

114 ERNEST C. FAUST

The external features of the worm and the digestive and reproductive

organs have been adequately studied, particularly by Stafford (1896).

From the material at my disposal I was able to develop certain facts re-

garding the excretor}'^ system which have previously been but poorly

understood.

The Excretory System in the Adult Worm

This was studied by StaflFord (1896: 506-516), who traced correctly

the main portions of the system, and followed "a few branches out to the

end organs." He states that "with this as a skeleton one can fill in mentally

the rest of the branches with their funnel organs and get a fair conception

of the semi-excretory system" (p. 511), From this study Stafford was able

to hypothecate the "tri-radiate order of branching" and a "tolerably clear

symmetry of arrangement." Although he was able to find a more exact

symmetry than Huxley (1856) and Voeltzkow (1888), he still found an

asymmetrical relation in the anterior branches on right and left sides. He

states further that "if all the branchings are regular and the process fs

repeated six times, we can, by a simple calculation, estimate the number of

funnel organs in a single Aspidogaster. Thus: 4iy^2)y,3'X?>y^3y.?) will

give the number for one side and doubling this we get a total of 1944."

The apparent asymmetry which Stafford found in the anterior branches

is not substantiated by my studies. Still more improbable is Stafford's

calculation of the number of ultimate capillaries based on the study of

"a few branches followed out to the end organs." Moreover, the irregu-

larities which he notes in other parts of the system can only be interpreted

as inadequate analysis. This is particularly true in the light of my studies

which indicate complete symmetry and regularity in the system of the larva

as well as in that of the adult.

While Stafford predicates four main branches, each of which proceeds to

trifurcate successively five times, I have found only three such main stems

and each of these in turn branches trichotomously only four times. Each

of these branches has been traced to its distal termini, with the result that

complete regularity is found to obtain (Fig. 1). In other words, the system

for each side of the body actually consists of 3 stems, each branching four

times, with 81 terminal units, giving a total of 243 capillaries and flame-cells

for each side of the body. This, as against Stafford's mental calculation

of 972 units which is theoretical and obviously based on inadequate analysis.

The system may be expressed as (3X3X3X3)-f(3X3X3X3X) +

(3X3X3X3) or (3)*+(3)''+(3)^ and reduces to the formula a"+i(3" + 7".

These data are supported further by the elemental excretory system

which is found in the larva (figs. 5, 6), Here three primitive flame-cells

and capillaries are seen which are the basal units of the system, (a-\-0-\-y).

These by successive trichotomies give rise to the adult system.

NOTES ON THE EXCRETORY SYSTEM 115

It seems desirable, in passing, to note that the larva (fig. 7) has paired

cluster of cephalic glands (eg), emptying through a cord of cephalic ducts

(cgd) just anterior to the oral sucker. This is analagous to the salivary

glands described for the redia of Cercaria equitator (Ssinitzin 1911: 52,

fig. 50) and C. flabellijormis (Faust 1918: 34, fig. 4v3) and might lend sup-

port to the view that the aspidobothrid is a redia with germinal epithelium

highly differentiated.

Summary

1. Record is made of the presence in several centers in China of the

cosmopolitan worm, Aspidogaster conchicola. In addition to the usual

hosts, Amyda sinensis has been found to harbor it.

2. The excretory system of the worm is regular and bilaterally symme-

trical. It has three main stems on each side of the body, each stem having

a 4-fold trichotomy.

3. This is expressed as (3X3X3X3) + (3X3X3X3) + (3X3X3X3)

or (3)^+ (3)^+ (3)^ and may be reduced to the formula a"+i8"+7".

4. The fundamental a-\-^-\-y pattern obtains in the larva where each

element is represented by a single flame-cell.

5. The presence of cephalic glands in the larva may support the view

that the worm is homologous to a highly differentiated redia.

Literature Cited

Faust E. C. 1918. Life History Studies on Montana Trematodes. 111. Biol. Monogr.,

4:1-21, 9 pi.

Huxley, T. 1856. Lectures on General Natural History. Med. Times and Gaz. London,

13:131-134, figs. 1-7.

Ssinitizin, D. Th. 1911. La generation parthenogenetique des Trematodes et sa descendence

dans les mollusque de la Mer Noire. Mem. Acad. Sci. St. Petersbourg, (8) 30:1-127,

6 pi. Russian.

Stafford, J. 1896. Anatomical Structure of Aspidogaster conchicola. Zool. Jahrb., Anat.,

9:476-542, 4 pi.

VoELTZKOW, A. 1888. Aspidogaster limacoides. Arb. Zool-Zoot. Inst. Wiirtzb. Wiesb.

8:290-292.

Key to Figures

o, anterior fundamental of excretory system.

8, median fundament of excretory system.

7, posterior fundament of excretory system.

b, bi, bii, bladder.

eg, cephalid gland.

cgd, cephalic gland duct.

ct, primary collecting tube.

ep, epi, epii, excretor>' pore.

rt, reflexed (secondary') collecting tubule.

116

ERNEST C. FAUST

Fig. 1

PLATE Xlir

Description of Figures

Fig. 1 . Lateral view of adult A spidogaslcr conchicola, showing complete excretory system

for right side of body, X 75.

NOTES ON THE EXCRETORY SYSTEM

117

Figs. 2 to 4. Stages in the escape of the larva from the egg shell, X 360.

Figs. 5, 6. Lateral and ventral views of the free larva, showing fundaments of the ex-

cretory system. The two sides are seen to develop separately, even to the e.xcretory pores.

X J(JO.

Fig. 7. View of larva showing cephalic glands and ducts, X 360.

A NEW CESTODE FROM LIPARIS LIPARIS*

By

Edwin Linton

University of Missotiri

The name Spathebothrium simplex gen. et sp. nov. is proposed for a

cestode collected at Woods Hole, Mass., by the late A'^inal N. Edwards

from the sea snail, Liparis liparis.

Mr. Edwards's record is as follows:

March 25, 1904, 2 fish examined, stomachs filled with small sand fleas,

2 tape worms in intestine of one.

April 14, 1904, 15 fish examined, stomachs filled with sand fleas; full of

spawn, nearly ripe; 7 tape worms from 4 fish.

January 14, 1905, 2 fish examined, one tape worm in each.

The cestodes in these three lots belong to the same species. Their

lengths, in alcohol, were 12, 14, 15, 16, 18, 18, 20, 20, 21, 26 and ic> milli-

meters respectively. The maximum breadth was about 2.25 millimeters;

ova 0.036 by 0.021 mm. in the two principal diameters.

The strobile is flattened, nearly linear, bluntly and smoothly rounded

at the two extremities. About" the only difference noted between the

anterior and posterior ends, as seen in whole mounts, is that genitalia are

wanting for a short distance at the anterior end while the vitellaria continue

to the extreme tip of the posterior end. In a specimen 11 mm in length

the first cirrus is 0. 44 mm. from the anterior end. The scolex therefore is

represented by the short portion which precedes the genitalia and is

probably transparent in life. In this specimen the vitellaria began a little

posterior to the level of the cirrus.

The strobile is not divided into distinct proglottides, the only indication

of strobilation being the successive sets of genital apertures, and, in cleared

and mounted specimens, the ovaries which are conspicuous, lobed, and

lie between the genital apertures at what would be the posterior end of a

proglottis, if proglottides were present.

The reproductive apertures are situated along the median line and are

not restricted to one of the flat surfaces of the strobile. For example, in a

specimen which had 20 s^ets of reproductive organs twelve of these o])ened

on one of the flat surfaces of the strobile and eight on the other. In another

specimen sixteen were counted on one side and nine on the other. In a

specimen 16 mm. in length the distance between adjacent sets of reproduc-

tive pores was about 0.67 mm., the first set lying about the same distance

from the anterior end. The apertures of the cirrus, vagina and uterus are

*Contribution from the U. S. Biological Station, Wtxwls Hole, Mass., and the Zoological

Laborator>- of the University of Missouri.

118

A NEW CESTODE FROM LIPARIS 119

4

near together, that of the cirrus being a little anterior to those of the

uterus and vagina, which are very near together and about at the same level.

The vitellaria continue without interruption from near the anterior

end to the posterior end, so that the strobile superficially resembles an

elongated trematode. The testes lie for the most part in front of the ovary

and are medially placed with respect to the vitellaria. In transverse sec-

tions through regions where the uterus is filled with ova the testes are

lateral and near the vitellaria (Fig. 6). The cirrus pouch is short but with

relatively thick muscular walls. The vagina has a strong muscular sphinc-

ter near its external opening (Figures 2, 4, 5). In ripe strobiles each set of

reproductive apertures is preceded by a mass of ova.

The musculature, so far as it is shown in sections, is poorly developed.

A few longitudinal fibers were noted in the subcuticula, but no trace of a

layer of longitudinal fibers between the subcuticula and central parenchyma

was seen, nor was there any indication of a circular layer. The cuticle

(Fig. 3) consists of two layers, an outer made up of short rod-like structures,

and an inner structureless layer. The outer layer constitutes about two-

thirds of the thickness of the cuticle but it may be more or less abraded.

The subcuticula in my sections appears as a loose mesh of fine fibers with

scattering cells. The thickness of the cuticle in the section from which

figure 3 was sketched was 0:01 mm., of the subcuticula 0:07, and of the

smaller diameter of the section, representing the thickness of the strobile, 0.5.

Sections of the anterior end of a strobile show numerous anastomosing

vessels of the excretory system. These vessels were difficult to interpret in

transverse sections in regions of the strobile where the reproductive organs

had appeared. Nowhere were they satisfactorily seen to be definitely

established as dorsal and ventral lateral vessel. In cases where two prin-

cipal lateral vessels could be distinguished they lay in about the same hori-

zontal plane with reference to the axis of the strobile. From a study of a

series of sagittal sections the lateral vessels were interpreted to be two,

with thin walls, somewhat tortuous, and giving off transverse branches.

This cestode is peculiar in the absence of bothria, and in certain char-

acteristics of the genital pores. The three genital apertures, cirrus,

uterus, and vagina, are, as a rule, near together on the median line, and

irregularly alternate with respect to the so-called dorsal and ventral surfaces

of the strobile. This feature stands in the way of referring it to the Pseudo-

phyllidae, which group is characterized by having the opening of the uterus

always on one of the faces, although the openings of the cirrus and vagina

may stand on opposite faces, or on a lateral margin.

// is thus seen that the species with which we are dealing is unique in that

it is not possible to speak of a dorsal and ventral surface of the strobile. For it

will be observed that, not only are the reproductive apertures irregularly alter-

nate on the flat surfaces of the strobile, but the reproductive organs themselves

are also irregularly alternate with respect to those surfaces (Fig. 2).

120 EDWIN LINTON

While examining a large number of transverse sections a single excep-

tional disposition of the reproductive apertures was noted. Figures 7 and

8 are sketches of this exceptional condition. Here the aperture of the cirrus

is seen to be on one of the flat surfaces of the strobile while the openings of

the vagina and uterus are on the opposite side. Since the apertures of the

uterus and vagina do not lie in the same horizontal plane it was necessary

to make two sketches. In the series of sections in which this anomalous

condition was noted two sections intervened between the sections shown in

figures 7 and 8.

An interesting feature with respect to the relative position of ovary and

vaginal aperture is shown in figure 2. In the upper part of the figure the

ovary is seen to be on the opposite side of the strobile from the vaginal

aperture, in the lower part of the figure it is on the same side. In the

former case the vagina crosses from one side of the strobile to the other, in

the latter it turns abruptly posteriad near the aperture.

Synopsis of genus Spathehothrium

No distinct scolex; strobile taenaeiform, bluntly rounded at the extremi-

ties, proglottides not distinct, reproductive apertures on median line and

irregularly alternate.

Explanation of Plate

c. cirrus sr. seminal receptacle

cp. cirrus pouch /. testes

cu. cuticle u. uterus

m. sphincter muscle of vagina v. vagina

0. ovary vd. vas deferens

sc. subcuticula vg. vitelline glands.

sg. shell gland

Fig. 1. Sketch of specimen mounted in balsam, somewhat diagrammatic. In this

specimen there were 12 sets of reproductive apertures on one side and 8 on the other. Length

16 mm.

Fig. 2. Sagittal section near median line showing reproductive apertures on opposite

sides of the strobile. The succeeding section to this in the series shows the uterus in about the

same relative position as that of the vagina in the lower left of the sketch. Thickness of

strobile at this point 0.30 mm.

Fig. 3. Cuticle and subcuticula highly magnified. Thickness of cuticle 0.01 mm.

Fig. 4. Reproductive apertures as seen in horizontal section. Sketch made from section

showing first appearance of the uterus. The vagina had appeared first in the preceding section,

and the cirrus in the si.xth preceding section. Diameter of cirrus bulb 0.24 mm.

Fig. 5. External apertures of vagina and uterus, transverse section. Long diameter of

ovum 0.035 mm.

Fig. 6. Transverse section showing uterus with ova, etc. Breadth of strobile at this

point. 1.5 mm.

Fig. 7. Transverse section showing e.xccptional arrangement of genital pores, the cirrus

opening on one side and the vagina on the other. Longer diameter of section 0.98 mm.

Fig. 8. From same series of sections as Fig. 7, two sections intervening between 7 and

8. The cirrus bulb still shows and the vagina is replaced by the uterus.

A NEW CESTODE FROM LIPARIS

121

PLATE XV

A LIST OF THE NEW GREGARINES DESCRIBED FROM

1911 TO 1920*

By

MiNNTE Watson Kamm

Although the gregarines are among the oldest known of the Protozoa

(Redi, 1684), they still remained a practically unknown group for a hundred

and fifty years, researches on more recently described groups far out-

numbering those on these parasites. This may have been due to the fact

that gregarines are of little or no economic importance. The hosts as a

rule are not animals of such import that the elimination of their parasites is

desirable and, moreover, the parasites themselves are generally harmless,

living commensal rather than actual parasitic lives within their hosts.

Because this is practically a new field, much of the work on the group

has been up to the present chiefly systematic; it is often easier to find an

entirely new species than to obtain a species already known. Considerable

has been done on Life Histories, Effect of the Parasite Upon the Host, and

Chromosome Behavior in the Complete Life-Cycle during the last decade

and much more is to be expected in these fields.

Labbe described the gregarines known up to the year 1899^ and reclass-

ified many of the wrongly designated and aberrent species. His paper

was probably the incentive for much of the subsequent work on the group.

The new forms described from the time of Labbe's Summary up to the year

1911 were listed by Sokolow^ and those described in this decade compared

favorably with the complete summary of Labbe.

Because the number of new species has been rapidly increasing subse-

quent to Sokolow's List, the writer has prepared a list of the species de-

scribed in the literature ivpm 1911 to the beginning of the year 1920. During

this decade were named many new genera and the genus Gregarina re-

ceived many new species.

Perhaps the most important researches of the decade were those in the

Suborder Schizogregarinae which includes many aberrent and apparently

unrelated species- and consequently the classification is considerably

confused.

The classification which follows is that of Minchin, at present the best

known.

* Contribution from the Zoological Laboratory of the University of Illinois, No. 204.

'Das Tierreich, Pt. 5, Sporozoa.

'Zool. Anz. 38:277-95; 304-14.

122

NEW GREGARINES DESCRIBED FROM 1911-1920 123

Class Sporozoa Leuckart

Subclass 1. Telosporidia Schaudinn 1900. Sporulation at end of

vegetative period.

Order 1. Gregarinoidea Minchin 1912. Trophozoite parasitic in

epithelial cells. Sporont free in a cavity. Spore

forms a single zygote.

Suborder 1. Eugregarinae Leger 1900. Reproduction by

sporogony only.

Tribe 1. Cephalina Delage and Herouard 1896. With

epimerite in trophozoite stage. Septate in all

but one family. Generally parasitic in digestive

tract of insects.

Tribe 2. Acephalina Delage and Herouard 1896. Without

epimerite, non-septate. Generally coelomic.

Suborder 2. Schizogregarinae Leger 1900. Reproduction by

both sporogony and schizogony.

Tribe 1. Monospora Leger and Duboscq 1908. Single

spore in sporogonic cycle.

Tribe 2. Polyspora Leger and Duboscq 1908. Many spores

in sporogonic cycle.

An Annotated List of Species in the Tribe Cephalina of the

Suborder Eugregarinae'

Family LECUNIDAE Kamm (1922).

Epimerite simple, symmetrical, gregarines non-septate, spores ovoidal,

thickened at one pole. Digestive tract of marine annelids.

Genus Lecudina Mingazzini 1891

Characters of the family

Lecudina sp.

Faria, Cunha, and Fonseca (1918) Mem. Inst. Osw. Cruz, 10:17-19.

Body spindle-shaped, nucleus spherical.

Host: Polydora social-is (Polych.) Taken at Rio de Janeiro, Brazil.

Family POLYRHABDINIDAE Kamm 1922.

Septate gregarines inhabiting the digestive tract of marine annelids.

Epimerites varied.

Genus Polyrhabdina Mingazzini 1891

(See Caullery and Mesnil 1914 C. R. Soc. Biol, 77:516-20.) Dicystid,

sporonts flattened, ovoidal, epimerite a corona of hooks. Intestine of

polychaetes of the family Spionidae.

Polyrhabdina spionis (Kolliker) (New name for Gregarina spionis Koll.)

^ All parasites described are intestinal forms unless otherwise stated.

124 MINNIE W. KAMM

Type species. Caullery and Mesnil (1914) C. R. Soc. Biol., 77. 516-20.

Host: Scololepsis fnliginosa. (Polych.)

Polyrhabdina polydorae (Leger) (New name for Doliocystis p. Leger.)

Caullery and Mesnil (1914), C. R. Soc, Biol., 77:516-20.

Host: Polydora ciliata. (Polych.)

Polyrhabdina brasili

Caullery and Mesnil (1914) C. R. Soc. Biol., 77:516-20.

Spor. ovoidal, 1. 200;u. Epim. like type, spines shorter.

Host: Spio martinensis. (Polych.)

Polyrhabdina pygospionis

Caullery and Mesnil (1914) C. R. Soc. Biol., 77:516-20.

Host: Pygospionis seticornis. (Polych.)

Family CEPHALOIDOPHORIDAE Kamm 1922 (this paper)

Characters of the type genus

Genus Cephaloidophora Mawrodiadi 1908

{ = Frenzelina Leger and Duboscq 1907, preocc. See Arch. zool.

exper., 46:lix-lxx.) Sporonts biassociative, no epimerite, cyst dehiscence by

simple rupture, spores ovoidal with equatorial line. Development intra-

cellular. Parasites of Crustacea.

Cephaloidophora maculata

Leger and Duboscq (1911) Arch. zool. exper., 46:lix-lxx.

Spor. ovoidal, max. 1. 80/i. Nucl. spher. cysts spher. 100/z, spores spher. 4/1.

Host: Gammarus marinus. (Crust.) Taken at Roscoff, France.

Cephaloidophora talitri

Mercier (1912) C. R. Soc. Biol., 72:38-9.

Spor. ovoidal, average 1. 40//, nucl. spher.

Host: Talitrus saltator. (Crust.) Taken at Roscoff, France.

Cephaloidophora ( = Frenzelina) delphinia^

Watson (1916) Jour. Parasit., 2:129-35.

Spor. ovoidal, largest spor. 115juX64ai. LP:TL::1:4; WP:WD::1:1.5.*

Nucl. spher. Cysts spher. SOfx.

Host: Talorchestia longicornis. (Crust.) Taken at Cold Spring Harbor,

L. I.

Cephaloidophora {^Frenzelina) olivia

Watson (1916) Jour. Parasit., 2:129-35.

Spor. ellipsoidal, largest 118^X36^. LP:TL::1:5; WP:WD::1 :1.3.

♦ The ratios of length of protomcrite to total Icnj^lh of sporont and width of protomerite

to width of deutomerite are given for average individuals. These ratios will be abbreviated

as above subsequently. Many recurring words will also be abbreviated from now on.

NEW GREGARINES DESCRIBED FROM 1911-1920 125

Cysts spher., 60/i.

Host: Libinia dubia. (Crust.) Taken at Cold Spring Harbor, L. I.

CephaloidopJwra ( = Frenzelina) nigrofusca

Watson (1916) Jour. Parasit., 2:129-35.

Spor. ovoidal, largest 125/iX75m. LP:TL::1:4; WP:WD::1:1.5. Nucl.

spher.

Hosts: Uca pugnax, U. pugilator. (Crust.) Taken at Cold Spring Har-

bor, L. I.

Cephaloidophora ( = Frenzelina) ampelisca

Nowlin and Smith (1917) Jour Parasit., 4:83-88.

Spor. 62/xX15m. Chromidial body in protomerite.

Host: Ampelisca s pint pes. (Crust.) Taken at Woods Hole, Mass.

Family STENOPHORIDAE Leger and Duboscq 1904. Spor. solitary,

Intracellular development. Dehiscence by simple rupture, spores ovoidal

with equatorial line. Epimente absent or rudimentary. Parasites of Diplo-

poda.

Genus Stenophora Labbe 1899

With the characters of the family

Stenophora elongata

Ellis (1912) Zool. Anz., 39:685-6.

Spor. elongate-cylindr., max. 1. 390/x. LP:TL::1:20; WP:WD::1:1 to

1:1.6. Prot. pentagonal.

Host: Orthomorpha coarctata. (Dipl.) Taken at Quirigua, Guatemala.

Stenophora cockerellae

Ellis (1912) Zool. Anz., 39:681-5.

Spor. elongate-cylindr., max. 1. 850/1. LP:TL::1:15; WP:WD::1:1.7.

Host: Parajulus sp. (Dipl.) Taken at Quirigua, Guatemala.

Stenophora robusta

Ellis (1912) Zool. Anz., 40:8-11.

Spor. short, avg. 1, 140-180//, w. 67^. LP:TL::1:8; WP:WD::1:2.5.

Hosts: Parajulus venustus; Orthomorpha gracilis; O. sp. (Dilp.) Taken

at Boulder, Col.

Stenophora impressa

Watson (1915) Jour. Parasit., 2:29; (1916) 111. Biol. Monogr., 2:280.

Spor. ellipsoidal, largest 375/iX48/i. LP:TL::1:12; WP:WD::1:2.3.

Cysts spher. 160/t.

Host: Parajulus impressus. (Dipl.) Taken at Urbana, 111.

Stenophora diplocorpa

Watson (1915) Jour. Parasit., 2:29; (1916) 111. Biol. Monogr., 2:284.

126 MINNIE W. KAMM

Spor. elongate-cylindr., constricted in mid-deut. LP:TL::1:20; WP:WD::

1:2.

Host: Eiiryurus crythropygus. (Dipl.) Taken at Urbana, 111.

Stenophora lactarla

Watson (1915) Jour. Parasit., 2:29; (1916) 111. Biol. Monogr., 2:282.

Spor. elongate-ellips , largest 480juX39yu LP:TL::1:12; WP:WD::1 :1 2.

Cysts spher. 170^1.

Host: Calliptis lactarius (Dipl.) Taken at Urbana, 111.

Stenophora caudata { = Spirosoma caud.)

Ishii (1915) Ann. zool. japon., 9:7-9.

Spor. tadpole-like in shape, posterior half reduced to cylindrical 'tail'

knobbed at end and spirally striated. Max. 1. 400;/, max. w. 100^.

LP:TL::1:12. Prot. papillate at apex.

Host: Fontaneria coarctata Pocock. (Dipl.) Taken in Gifu, Japan.

(The new genus Spirosoma Ishii is named from the spiral deutomerite,

none of the generic characters — epimerite, cystdehiscence, spores — being

found. From the few positive characters — shape of sporont, protomerite,

and diplopod host — it appears to belong to Stenophora. The author's

specimens were described from alcoholic specimens only.)

Stenophora cunhai

Pinto (1918) Brazil-Medico; (1919) Contribuifao ao estudo das

Gregarinas, Rio de Janeiro, 116 pp., 6 pi.

Spor. elongate-cylindr., prot. sub-spher., largest spor. 250/iX40/i. LP:TL::

1:5; WP:WD::1:1. Nucl. spher., in post, part of deut.

Host: Rhinocricus pugio. (Dipl.) Taken at Rio de Janeiro, Brazil.

Stenophora lutzi

Pinto (1918) Brazil-Medico; (1919) Contribuifao ao estudo das

Gregarinas, Rio de Janeiro, 116 pp., 6 pi.

Spor. elongate-cylindr. prot. cylindr. with constriction below middle.

Largest spor. 210;uX15a(. LP:TL::1 :7.5; WP:WD::1:1.2. Nucl. small,

spher.

Host: Rhinocricus sp. (Dipl.) Taken at Rio de Janeiro, Brazil

Stenophora cruzi

Pinto (1918) Brazil-Medico; (1919) Contribuijao ao estudo das Gre-

garinas, Rio de Janeiro, 116 pp., 6 pi.

Spor. elongate-cylindr. conical posteriorly, largest ^spor. [400^1 X 30m.

LP:TL::1:13; WP:WD::1:2. Prot. a truncate cone. Nucl. unknown.

Host: Rhinocricus sp (Dii)l.) Taken at Rio dc Janeiro, Brazil.

NEW GREGARINES DESCRIBED FROM 1911-1920 127

Stephora viannai

Pinto (1918) Brazil-Medico; (1919) Contribuiyao ao estudo das Gre-

garinas, Rio de Janiero, 116 pp., 6 pi.

Spor. stout-cylindr., bluntly conical posteriorly, largest spor. 1000//X150^(.

LP:TL::1:16; WP:WD::1:2. Nucl. elongate-cylindr.

Host: Rhinocricus sp. (Dipl.) Taken at Rio de Janeiro, Brazil.

Stenophora umbilicata

Pinto (1918) Brazil-Medico; (1919) Contribui(jao ao estudo das

Gregarinas, Rio de Janeiro, 116 pp., 6 pi.

Spor. stout-bodied, ovoidal, prot. small, broad. Hat. 320yuX 130//. LP:TL::

1:6; WP:WD::1:3.7. Nucl. spher.

Host: Rhinocricus sp. (Dipl.) Taken at Rio de Janeiro, Brazil.

Stenophora tenuicolUs

Pinto (1918) Brazil-Medico; (1919) Contribui^ao ao estudo das

Gregarinas, Rio de Janeiro, 116 pp., 6pl.

Spor. elongated globe-shaped in ant. fourth of deul. constricted to a 'wasp-

waist' and widening gradually toward post, end, end broadly-rounded,

prot. elongate-conical. Nucl. small, spher. Sporont 400;uX50/x.

Host: Rhinocricus sp. (Dipl.) Taken at Manguinhos, Rio de Janeiro,

Brazil.

Genus Fonsecaia Pinto (1918)

Brazil-Medico; (1919) Contribuijao ao estudo das Gregarinas, Rio de

Janeiro, 116 pp., 6 pi. Like Stenophora except spores elongate-ellipsoidal,

no endospore. Epimerite simple, without protoplasm. (The differentiation

of this genus from Stenophora is not convincing.)

Fonsecaia polymorpha. Type species

Pinto (1918) Brazil-Medico; (1919) Contribuigao ao estudo das

Gregarinas, Rio de Janeiro, 116 pp., 6 pi.

Spor. 170/iX80/i. Broadly ovoidal, prot. small, conical. LP:TL::1:11.3;

WP:WD::1:4.4. Nucl. spher. Spores ovoidal 18^^X8//.

Host: Orthomorpha gracilis. (Dipl.) Taken at Rio de Janeiro, Brazil.

Family GREGARINIDAE Labbe 1899

Epimerite symmetrical. Sporonts associative or solitary. Cysts with

or without spore-ducts.

Genus Gregarina Dufour 1828

Biassociative in sporont stage. Epimerite globular or cylindrical.

Spores regular. Cysts with spore-ducts.

Gregarina ctenocephalus { = G. ctenocephalus canis)

Ross (1909) Ann. Trop. Med. Par., 2:359-63.

Spor. spherical, no dimensions given. Epimerite pyriform, spores barrel-

shaped.

128 MINNIE W. KAMM

Host: Ctenoccphalus serraticeps (Acarinidae.) Taken at Port Said, Egypt.

Omitted from Sokolow's List.)

Gregarina creda

Wellmer (1911) Schr. Physik. Ges. Konigsbg., 52:115-6.

Spor. elongate-cylindrical, largest spor. 730yuX60/x. LP:TL::1:5; WP:WD

::1:1. Nucl. spher., cysts spher., SOOfx, spores typical, 6.4/iX3.2/i.

Host: Broscus cephalotes. (Col.) Taken in East Prussia.

Gregarina ovoid ea

Wellmer (1911) Schr. Physik. Ges. Konigsbg., 52:117.

Spor. obese, max. 1. 200ju. LP:TL::1:5; WP:WD::1:1.8. Nucl. spher.

Cyst spher. ISO/i.

Host: Crypticus quisquilins. (Col.) Taken in East Prussia.

Gregarina polyaulia

Wellmer (1911) Schr. Physik. Ges. Konigsbg., 52:118-9.

Spor. cylindr., largest spor. 470^X250^. LP:TL::1:6; WP:WD::1:1.8.

Cysts spher., 450/i, spores typical, 8.2^tX3.8;u.

Hosts: Harpalus aeneus and H. ruficornis. (Col.) Taken in East Prussia.

Gregarina rostrata

Wellmer (1911) Schr. Physik. Ges. Konigsbg., 52:120-1.

Spor: elongate-ovoidal, largest spor. 200/i long. LP:TL::1:7; WP:WD::

1:2. Nucl. spher., epimerite elongate-cylindrical. Cysts spher., 205/x,

spores ovoidal, 5.6juX3.2/i.

Host: Lagria hirta. (Col.) Taken in East Prussia.

Gregarina (Gigaductus) exiguus

Wellmer (1911) Schr. Physik. Ges. Konigsbg., 52:121-2.

Spor. obese, max. length 75m. LP:TL::1:4; WP:WD::1:2. Cysts spher.,

35m, one long spore-duct. Spores cylindr., 11.3mX4.8m.

Host: Pterostichus niger. (Col.) Taken in East Prussia. The genus

Gigaductus has been dropped. See Watson (1916) 111. Biol. Monogr., 2:

317, 389.

Gregarina guatemalensis

Ellis (1912) Zool. Anz., 39:687-8.

Spor. somewhat rectangular, max. 1. 276/x. LP:TL::1:3; WP:WD::1:1.5

Nucl. spher.

Host: Ninus inter stitialis. (Col.) Taken at Quirigua, Guatemala.

Gregarina consohrina

Ellis (1913) Trans. Amer. Micr. Soc, 32:267.

Spor. obese, average sporont 600m 1., 450m, w. LP:TL::1:5; WP:WD::

1:1.5. Cysts spher., 300m. Sporeducts up to 1200m in 1. Spores i-lnX^n.

Host: Ceuthophilus valgus. (Orth.) Taken near Boulder, Colo.

NEW GREGARINES DESCRIBED FROM 1911-1920 129

Gregarina grisea

Ellis (1913) Zool. Anz., 42:200.

Spor. ellipsoidal, max. 1. 540/z. LP:TL::1:4; WP:WD::1:1.1. Nucl. spher.

Host: Tcnehrio castaneus. (Col.) Taken at New Orleans, La.

Gregarina longiducta

Ellis (1913) Zool. Anz., 43:78-82.

Spor. obese, associations avg. 800-900^1 in 1. LP:TL::1:3; WP:WD::1:1.

Cysts spher., 560/x. Spores 3aiX6.5^(.

Hosts: Ceuthopilus latcns,C. maculatus. (Orth.) Taken at Douglas Lake,

Mich.

Gregarina typographi

Fuchs (1915) Zool. Jahrb., Syst., 38:109-222.

Spor. stout-bodied, bluntly ovoidal. No measurements given. LP:TL::

about 1:3; WP:WD::1:1. Nucl. small, spher. Cysts spher., one large

spore-duct. Spores 34X22jU.

Host: Ips typographus. (Col.) Taken in Southern Germany.

Gregarina { = Clepsidrina) hylohii

Fuchs (1915) Zool. Jahrb., Syst., 38:109-222.

Spor. long-ellipsoidal, largest spor. 847/iX304/z. Nucl. elongate-ellip-

soidal, one elongate karyosome. Cysts ovoidal 420 X370/x, without hyalin

envelope, spore-ducts numerous, spores rectangular with spine at each

corner, 6X4^.

Host: Hylobius ahieies. (Col.) Taken in Southern Germany.

Gregarina niinnta

Ishii (1914) Ann. Zool. japon, 8:436-8; Watson (1916) 111. Biol. Monogr.

2:343, 392, 409.

Spor. elongate-cylindr., assn. 1. 118/x. LP:TL::1:9; WP:WD::1:1.7.

Nucl. spher., cysts spher., 48/^.

Host: Tribolium ferrugineum. (Col.) Taken in Prov. of Izu, Japan.

Gregarina globosa

Watson (1915) Jour. Parasit. 2:31; (1916) 111. Biol. Monogr., 2:401.

Spor. subglobose, 260^X180^. LP:TL::1:8.6; WP:WD::1:2.4. Nucl. spher.

Host: Coptotomus interrogatus. (Col.) Taken at Urbana, 111.

Gregarina monarchia

Watson (1915) Jour. Parasit., 2:31; (1916) 111. Biol. Monogr., 2:400.

Spor. elongate-cylindr., largest spor. 570/xXl30ju. LP:TL::1:7; WP:WD::

1:1.3.

Host: Pterostichus stygicus. (Col.) Taken at Urbana, 111.

130 MINNIE W. KAMM

Cregarina barbarara

Watson (1915) Jour. Parasit., 2:31; (1916) 111. Biol. Monogr., 2:394.

Spor. ovoidal, largest spor. 145^X90^. LP:TL::1:6; WP:WD::1:2. Nucl.

small, spher.

Host: Coccinella sp. (Col.) Taken at Oyster Bay, L. I.

Gregarina katherina

Watson (1915) Jour. Parasit., 2:31; (1916) 111. Biol. Monogr., 2:392.

Spor. ellipsoidal, largest spor. 78/iX35/x. LP:TL::1:7; WP:WD::1:1.7.

Nucl. spher.

Host: Coccinella novemnotata. (Col.) Taken at Oyster Bay, L. I.

Gregarina intestinalis

Watson (1915) Jour. Parasit., 2:32; (1916) 111 Biol. Monogr., 2:399.

Spor. broadly ellipsoidal, largest spor. 160/xX80/.t. LP:TL::1:5; WP:WD::

1:2.

Host: Pterostichus stygicus. (Col.) Taken at Urbana, 111.

Gregarina gracilis

Watson (1915) Jour. Parasit., 2:32; (1916) 111. Biol. Monogr., 2:398.

Spor. elongate-ellipsoidal, largest spor. 190aiX80/i. LP:TL::1:8; WP:WD::

1:2. Nucl. spher. cysts spher. 90/i.

Host: Larva of Elateridae. (Col.) Taken at Urbana, III.

Gregarina tenebrionella

Watson (1915) Jour. Parasit., 2:32; (1916) 111. Biol. Monogr., 2:397.

Spor. sub-globose, largest spor. 70/iX42M. LP:TL::1:4; WP:WD::1:1.7.

Nucl. spher.

Host: Larva of Tenebrionidae. (Col.) Taken at Urbana, 111.

Gregarina fragilis

Watson (1915) Jour. Parasit., 2:32; (1916) 111. Biol. Monogr., 2:395.

Spor. ellipsoidal, largest spor. 110^X60^:. LP:TL::1:5; WP:WD::1:2.

Nucl. spher.

Host: Coccinella sp. (Col.) Taken at Urbana, 111.

Gregarina nigra

Watson (1915) Jour. Parasit., 2:33; (1916) 111. Biol. Monogr., 2:326.

Spor. cylindrical, largest spor. 530aiX180m. LP:TL::1:4; WP:WD::

1:1.4. Nucl. spher.

Hosts: M elanoplus femur-rubrum , M. diferentialis, Encoptolophus sordidus.

(Orth.) Taken at Urbana, 111.

Gregarina galliveri

Watson (1915) Jour. Parasit., 2:33; (1916) 111. Biol. Monogr., 2:321.

NEW GREGARINES DESCRIBED FROM 1911-1920 131

Spor. 300aiX180//. LP:TL::1:5; WP:WD::1:1. Prot. flat, broad, deut.

widest in post. half. Nucl. spher. Cysts spher., 350//.

Host: Gryllus abbreviatus. (Orth.) Taken at Oyster Bay, L. I.

Gregarina stygia

Watson (1915) Jour. Parasit., 2:33; (1916) 111. Biol. Monogr., 2:324.

Spor. obese, largest 180/xXlOO/x. LP:TL::1:6; WP:WD::1:1.6. Nucl.

spher., cysts spher. 150/z.

Host: Ceuthophilus stygius. (Orth.) Taken at Cold Spring Harbor, L. I.

Gregarina illinensis

Watson (1915) Jour. Parasit., 2:34; (1916) 111. Biol. Monogr., 2:318.

Spor. elongate-cylindr., largest spor. 550yuXl80yu. LP:TL::1:5; WP:WD::

1:1.5. Nucl. small, spher.

Host: Ischnoptera pennsylvanica. (Orth.) Taken at Urbana, 111.

Gregarina platyni

Watson (1916) 111. Biol. Monogr., 2:402.

Spor. elongate-cylindr., max. 1. 610ai. LP:TL::1:4; WP:WD::1:1. Prot.

constricted in middle. Nucl. spher.

Host: Platynus ruficollis. (Col.) Taken at Oyster Bay, L. I.

Gregarina udeopsyllae

Watson (1916) 111. Biol. Monogr., 2:327.

Spor. obese, largest 310/xX200/i. LP:TL::1:5; WP:WD::1:1.5.

Host: Udeopsylla nigra. (Orth.) Taken at Urbana, 111.

Gregarina neglecia

Watson (1916) Jour. Parasit., 3:65-75.

Spor. ovoidal, largest spor. 500yuX230ju. LP:TL::1:6; WP:WD::1:1.5.

Cysts spher., 300/z.

Host: Ceuthophilus neglectus (Orth.) Taken at Oyster Bay, L. I.

Gregarina platydenia

Kamm (1918) Jour. Parasit., 4:159-63.

Spor. cylindr, slender, largest spor. 1210^X150^. LP:TL::1:12; WP:WD::

1:1.5. Nucl. spher. Epim. a simple cone.

Host: Platydenia excavatum. (Col.) Taken at Urbana, 111.

Gregarina diabrotica

Kamm (1918) Jour. Parasit., 4:159-63.

Spor. elongate-cylindr. largest spor. 270/xXl05/z. LP:TL::1:3.5; WP:WD::

1:1.6. Nucl. spher. Epim. a sessile knob.

Host: Diabrotica vittata. (Col.) Taken at Urbana, 111.

132 MINNIE W. KAMM

Gregarina watsoni

Pinto (1918) Brazil-Medico; (1919) Contribuifao ao estudo das Gre-

garinas, Rio de Janeiro, 116, pp.; 6 pi.

Spor. elongate-ovoidal, largest spor. 350mX152m. LP:TL::1:7; WP:WD::

1:1.5. Nucl. spher. Epim. globular.

Host: Omoplata nor?nalis. (Col.) Taken at Nictheroy, Brazil.

Gregarina chagasi

Pinto (1918) Brazil-Medico; (1919) Contribuifao ao estudo das Gre-

garinas, Rio de Janeiro, 116 pp., 6 pi.

Spor. sub-globular to cylindrical. Largest spor. ISO^iXSO^u. LP:TL::

1:3.6; WP:WD::1:1.5. Nucl. spher. Cysts ovoidal.

Host: Conocephalus f rater. (Orth.) Taken at Manguinhos, Brazil.

Gregarina aragaoi

Pinto (1918) Brazil-Medico; (1919) Contribuifao ao estudo das Gre-

garinas, Rio de Janeiro, 116 pp., 6 pi.

Spor. elongate-ovoidal, max. 1. 170/z, max. w. 70/i. LP:TL::1:5.7; WP:

WD::1:1.7. Nucl. spher. Epim. a small papilla. Cysts subspherical.

Host: Systena sp. (Col.) Taken at Manguinhos, Brazil.

Gregarina sp.

Wellmer (1911) Schr. Physik. Ges. Konigsbg., 52:146.

Host: Sviinthurns fuscus. (Thysan.) Taken in East Prussia.

Gregarina sp.

Wellmer (1911) Schr. Physik. Ges. Konigsbg., 52:148. '

Host: Oribata geniculata. (Arachn.) Taken in East Prussia.

Genus Hirmocystis Labbe 1899

Associations of two to twelve or more. Epimerite a small papilla.

Cysts dehisce by simple rupture. Spores ovoidal.

Hirmocystis harpali

Watson (1916) 111. Biol. Monogr., 2:378.

Spor. elongate, largest 500^X80/1. LP:TL::1:7; WP:WD::1:1.2. Max-

imum of four in a chain. Nucl. spher. Epim. large and spherical.

Host: Harpalus pennsylvanicus erythropus. (Col.) Taken at Urbana, 111.

Genus Uradiophora Mercicr 1912

Arch. zool. exper., (5) 10:198. Sporonts associative, cysts without

spore-ducts. Spores spherical or sub-spherical with equatorial line,

extruded in chains. Epimerite an elongated papilla. Deut. with small

appendix.

Uradiophora cuenoti { = Cephaloidophora cucnoti) Type species.

Mercier (1911) C. R. Soc. Biol., 71:51-3; (1912) Arch. zool. exper.,

(5) 10:177-202.

NEW GREGARINES DESCRIBED FROM 1911-1920 133

Spor. associated in chains of from 2 to 4 individuals, very elongate, max. 1.

spor. 700fx. Nucl. spher. Epim. an elongated papilla. Deut. with small

atrophied appendix. Cysts ovoidal, 44/i in 1., spores 4/i.

Host: Atyaephyra Desmaresti. (Crust.) Taken at Nancy, France.

Genus Pyxinoides Tregouboff 1912

Arch. zool. exper., (5) 10:liii-lxi. Sporonts in twos, development

extracellular, epimerite a slightly stalked globular papilla with 16 longi-

tudinal furrows, with small conical papilla at apex.

Pyxinoides balani. Type species

Tregouboff (1912) Arch. zool. exper., (5) 10:liii-lxi.

Max. I. primite 130;u, satellite 60^. Nucl. spher.

Hosts: Balaims amphitrite, B. eburneus. (Crust.) Taken at Cette, France.

Genus Leidyana Watson 1915

Jour. Parasit., 2:35. Sporonts solitary, epimerite a small sessile knob,

dehiscence by spore-ducts, spores in chains, dolioform.

Leidyana { = Stenophora) erralica. Type species.

Crawley (1903) Proc. Acad. Nat. Sci., Phila., 55:45.

Watson (1916) III. Biol. Monogr., 2:328-30.

Leidyana tinei

Keilin (1918) Parasit., 10:406-10.

Spor. long-ellipsoidal, max. I. 300At, w. 85^. LP:TL::1:1.5; WP:WD::1:

1.7. Cysts spher. llO/x, spores barrel-sh., 7/x long.

Host: Endrosis fenestrella. (Lepid.) Taken at Cambridge, Eng.

Genus Protomagalhdensia Pinto 1918, Brazil-Medico

Spores barrel-shaped with spine at each corner, sporonts attenuated,

several individuals in an association, often attached laterally. Myonemes

prominent.

Protomagalhaensia { = Gregarina) serpentula. Type species.

Magalhaes (1900) Arch, parasit., 3:34-69; Pinto (1918) Brazil-Medico.

Family DIDYMOPHYIDAE Leger 1892

Associations of two or three individuals. None-septate in satellites.

Genus Didymophyes Stein 1848

Epimerite a small pointed papilla. Cyst dehiscence by simple rupture.

Spores ellipsoidal.

Didymophyes { = Gregarina) minuta

Ishii (1914) Ann. Zool. japon., 8:435-41.

Watson (1916) 111. Biol. Monogr., 2:343.

Sporonts elongate, 188/^X26/^. Ratio LP:TL::1:23; WP:WD::1:1.5.

Nucleus spherical. Cyst and spores unknown.

Host: Tribolium ferrugineum. (Col.) Taken in Prov. of Izu, Japan.

134 MINNIE W. KAMM

Family ACTINOCEPHALIDAE Leger 1892

Sporonts solitary, epimerites varied, simple rupture of cysts.

Genus Actinocephalus Stein 1848

Epimerite with many upwardly-directed spines, spores biconical.

Actinocephalus permagnus (?.4. sp. Pfeiflfer 189vS; A. steUiformis Wasielew-

ski 1896)

Wellmer (1911) Schr. Physik. Ges. Konigsbg., 52:

Spor. elongate, max. 1. 1.3 mm. LP:TL::1:17; WP:WD::1.1 :1. Nucl.

ellipsoidal, cysts nearly spher., 750^t. Spores diamond-shaped, 7.6./iX5)U.

Host: Procrustes coriaceus. (Col.) Taken in East Prussia.

A ctinoccphalus parvus

Wellmer (1911) Schr. Physik. Ges. Konigsbg., 52:?.

Spor. ovoidal, largest 140^X75/^. LP:TL::1:5; WP:WD::1:1.3. Nucl.

ovoidal. Epim. a corona of digitiforn processes upon a short neck.

Hosts: CeraiophyUiis fringiUae, C. galUnae larv. (Dipt.) Taken in East

Prussia.

A ctinoccphalus echinatus

Wellmer (1911) Schr. Physik. Ges. Konigsbg., 52:?

Spor. cylindro-conical, largest 400m in 1. LP:TL::1:5; WP:WD::1.1 :1.

Cysts spher., 330^, spores biconical, 8^X4.8^.

Hosts: Pterostichus niger, P. vulgaris. (Col.) Taken in East Prussia.

Actinocephalus zophus { = Stephanophora zopJia Ellis (1913) Zool. Anz.,

42:200-2).

Ellis (1913) Trans. Amer. Micr. Soc, 32:278.

Spor. elongate-cylindr., max. 1. 1600^. LP:TL::1:12; WP:WD::1:1.7.

Epim. persistent, stout-necked, constricted at base, and terminating in

corona of 9 or more small digitiform processes.

Hosts: Xyctotheres barbarata {X. barbata), Alobates pennsylvanicus. (Col.)

Taken at New Orleans, La.

Actinocephalus brachydactylus

Ellis (1913) Trans. Amer. Micr. Soc, 32:279.

Spor. elongate-ovoidal, 1. 500^. LP:TL::1:4; WP:WD::1:1.

Host: Nymphs of Aeschna sp. (Neur.) Taken at Douglas Lake, Mich.

Actinocephalus crassus (^ = Stephana phora crassa Ellis (1912) Zool. Anz.,

39:688-9).

Ellis (1913) Trans. Amer. Micr. Soc, U:!!?^.

Avg. spor. 50m-60m in 1. LP:TL::1:3.5; WP:WD::1 :1.5. Nucl. spher.

Host: Leptochirus edax. (Col.) Taken at Quirigua, Guatemala.

Actinocephalus gimbeli { = Stenophora gimbeli Ellis (1913) Zool. Anz.,

41:464.) Watson (1916) III. Biol. Monogr., 2:353.

NEW GREGARINES DESCRIBED FROM 1911-1920 135

Spor. obese, 1. 500/x. LP:TL::1:5; WP:WD::1:1.2.

Host: Harpalus pennsylvanicus. (Col.) Taken at Vincennes, Ind.

Genus Pyxinia Hammerschmidt 1838

Epimerite a flat crenulate crateriform disc with central style. Spores

l)iconical.

Pyxinia bulhifera

Watson (1916) Jour. Parasit., 3:65-75.

Spor. long, slender, longest spor. 850/iXl60/i. LP:TL::1:5. WP:WD::

1:1.3. Epim. typical, 60m— lOO/i 1. Nucl. spher.

Host: Dermestes lardarius. (Col.) Taken at Oyster Bay, L. I.

Genus Amphorocephalus Ellis 1913

Zool. Anz., 41:462. Epim. dilated in middle, terminating in a concave

disc peripherally fluted at ant. end. Prot. constricted across middle. Spores

not known.

Amphorocephalus amphorellus. Type species.

Ellis (1913) Zool. Anz., 41:463-4; Trans. Amer. Micr. Soc, 32:276-7.

Spor. elongate, 1. 500m -970m. LP:TL::1:17; WP:WD::1:2.

Host: Scolopendra heros. (Chil.) Taken at Boulder, Col.

Genus Boihriopsis Schneider 1875

Epimerite with long slender filaments. Prot. very large. Spores

biconical.

Boihriopsis ( = Legeria) terpsichorella

Ellis (1913) Trans. Amer. Micr. Soc, 32:276; Watson (1916) 111. Biol.

Monogr., 2:356.

Prot. of spor. equal to or longer than deut. Avg. 1. 720m, w. 145m. LP:TL::

1.5:1; WP:WD::1.3:1.

Host: Hydrophilus sp. (Col.) Taken at Douglas Lake, Mich.

Boihriopsis claviformis

Pinto (1918) Brazil Medico; (1919) Contribuijao ao estudo das Gre-

garinas, Rio de Janeiro, 116 pp., 6 pi.

Spor. elongate-triangular, widest at ant. end, bluntly acuminate. LP:TL::

1:7; WP:WD::1.4 : 1.

Host: Aeschnida larva. (Odon.) Taken at Manguinhos, Brazil.

Boihriopsis osivaldocruzi

Hasselmann (1918) Brazil-Medico, Nov. 2, 1918.

Genus Siylocysiis Leger 1899

Epimerite a sharp recurved cone. Spores biconical.

Siylocysiis ensiferiis { = Siylocephalus en. Ellis 1912 Zool. Anz., 39:686)

Eliis (1913) Trans. Amer. Micr. Soc, 32:274.

Avg. 1. spor. 40-65m. LP:TL::1:2.5; WP:WD::1:1.2.

Host: Lepiochirus edax. (Col.) Taken at Quirigua, Guatemala.

136 MINNIE W. KAMM

Genus Steinina Leger and Duboscq 1904

Epimerite a short digitiform process changing into a flat button. Spores

biconical.

Steinina rotundata

Ashworth and Rettie (1912) Proc. Roy. Soc. Lond., B 86:31.

Spor. 180/i long, 80/i wide. Epim. sometimes a blunt cone with central

st3^1e, again a saucer-shaped disc with crenulate periphery. Spor. ovoidal,

nucl. spher. Cysts spher. 185/x, dehiscing in int. of host, spores ovoidal,

12aiX7/x.

Hosts: Ceratophylkis styx, C. farreui, C. galUnae. (Dipt.) Taken near

Edinburgh, Scotland.

Steinina obconica

Ishii (1914) Ann. zool. Japon., 8:439.

Spor. ovoidal, largest 148^X80^. LP:TL::1:5; WP:WD::1:1. Epim. a

minuted style. Prot. compressed ant. -post. Nuck spher. Cysts ovoidal.

Host: Tribolium ferrugineum. (Col.) Taken in Prov. of Izu, Japan.

Steinina rotunda

Watson (1915) Jour. Parasit., 2:32; (1916) 111. Biol. Monogr., 2:364.

Spor. globose, largest 250/^X130^. LP:TL::1:2.3; WP:WD::1:1.1. Epim.

spher.

Host: Amara angustata. (Col.) Taken at St. Joseph, 111.

Steinina harpali

Watson (1916) 111. Biol. Monogr., 2:365.

Coelomic. Spor. small, obese, largest spor. 200/iXlOOM. LP:TL::1:4;

WP:WD::1:1 .3. Epim. a short cone changing into a sphere then cup-

shaped. Nucl. small, spher. Cysts spher. 12/i.

Host: Harpalus pennsylvanicus longior. (Col.) Taken at Urbana, HI.

Family ACANTHOSPORIDAE Leger 1892

Spor. solitary, epim. varied. Dehiscence by simple rupture, spores with

equatorial and polar spines.

Genus Corycella Leger 1892

Epim. globular with 8 large recurved hooks, spores biconical, 4 spines at

each pole.

Corycella orthomorpha

Hasselmann (1918) Brazil-Medico, Oct. 5, 1918.

Genus Prismatospora Ellis 1914

Trans. Amer. Micr. Soc, 33:215. Spores hexagonal, truncate at ends

with one row of long spines at each pole. Epim. subglobose with lateral

recurved hooks.

NEW GREGARINES DESCRIBED FROM 1911-1920 137

Prismatospora evansi. Type species

Ellis (1914) Trans. Amer. Mic. Soc, 33:215.

Spor. 400/x in avg. 1., broadly conical, LP:TL::1:3; WP:WD::1:1; Prot.

broad, blunt, deut. tapering. Nucl. small, spher. Cysts subspher., 370yu,

dehiscence by simple rupture, spores ll/iX5.8/z

Hosts: Nymphs of Tramea lacerata and Sympetrum rubicunduliim. (Neur.)

Taken at Douglas Lake, Mich.

Genus Cometoides Labbe 1899

Epim. a sphere with long slender filaments. Spores biconical with one

polar and two equatorial rows of spines.

Cometoides sp.

Wellmer (1911) Schr. Physik. Ges. Konigsbg., 52:138.

Spor. cylindro-conical. Max. 1. 360/x. LP:TL::1:5; WP:WD::1:1. Nucl.

ellipsoidal. Epim. a flattened papilla with long filaments. Cysts spher.

160^.

.Host: Carabus sp. larva. (Col.) Taken in East Prussia.

Family STYLOCEPHALIDAE Ellis

New name for Stylorhynchidae Schneider 1886 preocc. Ellis (1912)

Zool. Anz., 39:25. Spor. solitary, epim. varied, nucl. ovoidal, dehiscence

by pseudocyst, spores irregular, in chains.

Genus Stylocephalus Ellis

New name for Stylorhynchus Stein 1848 preocc. Ellis (1912) Zool. Anz.,

39:25. Epim. a papilla at end of a long slender neck. Cysts papillate,

spores hat-shaped.

Stylocephalus giganteus

Ellis (1912) Zool. Anz., 39:25-7.

Spor. elongate, 1200-1800/1 in 1. LP:TL::1:15; WP:WD::1:1. Cysts

spher., 450/x. Spores 7 X 11m-

Hosts: Eleodes sp.; Asida opaca; Asida sp.; Eusattus sp. (Col.) Taken at

Boulder and Denver, Col.

Genus Bulbocephalus Watson 1916

Jour Parasit., 3:66. Epim. a dilated papilla in middle of rather long

slender neck. Nucl. ellipsoidal.

Bulbocephalus wardi. Type species

Watson (1916) Jour. Parasit., 3:66.

Spor. stout, widest at shoulder, largest spor. 290/zX45/i. LP:TL::1:5;

WP:WD::1:1. Epim. as above. Nucl. placed diagonally. Cysts and

spores unknown.

Host: Clerid larva. (Col.) Taken at Oyster Bay, L. I.

138 MINNIE W. KAMM

Bulbocephalus elongatus

Watson (1916) Jour. Parasit., 3:66.

Spor. very long and slender, max. 1. 600/z, w. 50/i. LP:TL::1:11; WP:WD::

1:1. Epim. as above. Nucl. diagonally placed.

Host: Cucujus larva. (Col.) Taken at Oyster Bay, L. I.

Family DACTYLOPHORIDAE Leger 1892

Epimerite complex, sporonts solitary, cysts dehisce by lateral pseudo-

cyst or simple rupture, spores elongate.

Genus Nina Grebnecki 1873

Protomerite two long lobes fused at one end, peripherally set with teeth

and long slender filaments. Spores in chains.

Nina indicia

Merton (1911) Abh. Seneck. nat. Ges. Frankf., 34:119-26.

Spor. elongate, max. 1. ISOO/x. LP:TL::1:20; WP:WD::4:1. Prot. low,

very broad, two long narrow parallel plates attached laterally, free at

one end, each plate armed with a ridge of short sharp teeth. Nucl. spher.

Host: Scolopendra subspinipcs. (Chil.) Taken at Heidelberg, Germ.

Nina ( = Pterocephalns) leitdodacimhai

Hasselmann (1918) Brazil-Medico, Sept. 21, 1918.

Genus Echinomera Labbe 1899

Epimerite an eccentric cone with short digitiform processes. Dehiscence

by simple rupture. Spores cyllindrical, in chains.

Echinomera ( = Gregarina) magalhaesi^

Pinto (1918) Brazil-Medico; (1919) Contribuifao ao estudo das Gre-

garinas, Rio de Janeiro, 116 pp., 6 pi.

Spor. elongate-conoidal, widest at shoulder. Largest spor. 300/zX70/i.

LP:TL::1:4.3; WP:WD::1:1 . 1. Epim. a polymorphic eccentric cone.

Nucl. ellipsoidal.

Host: Scolopendra sp. (Chil.) Taken at Rio de Janeiro, Brazil.

Genus Seticephaliis Kamm 1922 (this paper)

A dense tuft of short, upwardly-directed brush-like bristles super-

imposed upon the broad, flat-topped protomerite, persistent. A chromi-

dial body in protomerite. Parasitic in Chilopoda.

Seticephalus { = Gregarina) elegans. Type species.

Pinto (1918) Brazil-Medico; (1919) Contribuiffio ao estudo das Gre-

garinas, Rio de Janeiro, 116 pj)., 6 pi.

Spor elongate-conoidal, acuminate, largest spor. 75//X32"'. LP:TL::

1:7.5; WP:WD::1 :1 .2. Prot. broad, flat, nucl, small ellipsoidal. Epim.

* This species was placed by the author in the Genus Gregarina.

NEW GREGARINES DESCRIBED FROM 1911-1920 139

short bristle-like filaments across whole ant. end of prot. Cyst and spores

unknown.

Host: Scolopendra sp. (Chil.) Taken at Rio de Janeiro, Brazil.

(This species was placed by the author in the genus Gregarina but it is

unlike any member of that genus or any hitherto described genus and is

therefore made the type species of a new genus.)

Genera of Uncertain Position

Genus Agrippina Strickland 1912 Parasit., 5:108

Sporonts solitary, epim. a circular disc armed with peripheral digitiform

processes, with short neck. Spores long-ovoidal.

Agrippina bona. Type species

Strickland (1912) Parasit., 5:108.

Spor. elongate, conical, avg. 1. 175/i. Nucl. ellipsoidal. Epim. as stated.

Cysts spher. dehiscing by simple rupture. Spores 6.6/xX7)U-

Host: Ceratophyllus fasciatns. (Dipt.) Taken at Cambridge, England.

Genus Metamera Duke 1910

Q. J. Mic. Sci., 55:261-86. Epimerite subconical, apex eccentric, with

corona of numerous branched digitiform appendages. Cysts dehisce by

simple rupture. Sporonts solitary.

Metamera schubergi. Type species

Duke (1910) Q. J. Mic. Sci., 55:261-86.

Spor. 150/1 X45;u. Deut. with one to three septa posterior to nucleus.

Cysts spher., spores ovoidal, 9/^X7 /jl.

Hosts: Glossophonia complanata, Hemiclepsis marginata. (Annelida.)

Taken at Heidelberg, Germ, and Cambridge, Eng.

(This species was left out of Sokolow's Synopsis (1911).)

Genus Ganymedes Huxley 1910

Q. J. Mic. Sci., 55:155-75. Non-septate, with motile extensile fixation-

organ, cupped posterior end for association, nucleus large, spherical.

Inhabit intestine and liver of host.

Ganymedes anasidis

Huxley (1910) Q. J. Mic. Sci., 55:155-75.

Characters of the genus. Avg. 1. 250-300/1, w. 17-2b/t. Spor. elongate-

cylindrical.

Host: Anaspides tasmaniae. (Crust.) Taken in Tasmania.

(This species was omitted from Sokolow's List — 1911.)

Species of Uncertain Position

Gregarina crassa

Ishii (1915) Ann. zool. japon., 8:438-9.

Spor. ovoidal, max. 1. 242/1, w. 64/i. Nucl. spher. LP:TL::1:19; WP:WD::

1 :4.

140 MINNIE W. KAMM

Host: Triholium jerrugmeum. (Col.) Taken in Prov. of Izu, Japan.

Prot. lacking in satellite. See Watson (1916) 111. Biol. Monogr., 2:409.

Gregarina coptotomi

Watson (1916) Jour. Parasit., 2:406.

Spor. solitary, epim. and cysts unknown. Spor. 210/i 1. LP:TL::1:7;

WP:WD::1:2.3. Nucl. ellipsoidal.

Host: Co ptotomus interrogatus. (Col.) Taken at Urbana, 111.

Gregarina brasUiensis

Pinto (1918) Brazil-Medico; (1919). Contribuicao ao estudo dos

Gregarinos Rio de Janeiro, 116 pp., 6 pi.

Spor. not associative, pyriform, acutely acuminate, largest spor. 92/xX35/i.

LP:TL::1:2.4; WP:WD::1:1.1. Prot. ovo-cylindrical, nucl. ovoidal.

Host: Scolopendra sp. (Chil.) Taken at Rio de Janeiro, Brazil.

Gregarina legeri

Pinto (1918) Brazil-Medico; (1919). Contribuicao ao estudo dos

Gregarinos Rio de Janeiro, 116 pp., 6 pi.

Spor. not associative, rectangular with bulbous post, extremity, largest

spor. 290/zX80/i (at post, end of deut.) LP:TL::1:4.8; WP:WD::1:1.

Prot. square, nucl. ellipsoidal, in dilated post, portion of deut.

Host: Stylopyga americana. (Orth.) Taken at Rio de Janeiro, Brazil.

Taeniocystis legeri

Cognetti de Martiis (1911) Arch. Protistenk., 23:247.

Spor. segmented in both prot. and deut., max. 1. 1600/x. Up to 19 segments.

Nucl. ovoidal. Epim., cysts, and spores unknown.

Host: Kynotus PittarelUi (Oligoch.) Taken at Moramanga, Madagascar.

This species is placed among the 'Uncertain Species' because the

'protomerite' is divided into three segments and the parasite is coelomic.

Miscellaneous

Unnamed gregarines:

Wellmer (1911) Schr. Physik. Ges. Konigsbg., 52:46-7. From the fol-

lowing hosts: Heledona agricola (Col.), Polyporus sulphureiis (Plathy.),

Tritoma quadripitstulata (Col.), Cychrus rostratiis (Col.), Scolopcndrclla

sp. (Chil.).

Unnamed Cometoides-like form:

Wellmer (1911) Schr. Physik. Ges. Konigsbg., 52:46-7. Host: Ilydro-

philus aterrimus. (Col.).

Unnamed gregarines of several species:

Pantel (1913) La Cellule, 29 (1): 142-4. Host: Forficula auricularia.

(Orth.)

NEW GREGARINES DESCRIBED FROM 1911-1920 141

Unnamed gregarine:

Buddington (1910) Science, 31:470. Host: Balanus ebiirneus. (Crust.)

Unnamed gregarines similar to Leidyana tinei:

Keilin (1918) Parasit., 10:406. Hosts: Oecophora pseudospretella,

Tinea pallescentella. (Lepidopt.)

An Annotated List of Species in the Tribe Acephalina of the

Suborder Eugregarinae

Genus Monocystis Stein 1848

Non-septate, irregular, motile sporonts, cysts with incomplete sporu-

lation, spores navicular, octozoic. (All herein described are coelomic or

inhabit seminal vesicles unless stated otherwise.)

Monocystis pareudrili

Cognetti de Martiis (1911) Arch. Protistenk., 23:216-40.

Spor. subspherical, max. diam. 60;u. Spores ovoidal, 10X5/z.

Host: Pareiidrilus pallidus. (Polych.) Taken in 'Equatorial Africa.'

Monocystis thamnodrili { = M. sp. Cognetti 1906)

Cognetti de Martiis (1911) Mem. R. Accad. Sci. Torino, 46: (2) 147-262.

Host: Rhinpdrilus {=21iamnodrilus) incertus. (Polych.) Taken in

Ecuador.

Monocystis rostrata

Muslow (1911) Arch. Protistenk., 22:20-55.

Sem. ves. Spor. spindle-shaped, cysts spher., spores spindle-shaped.

Host: Lumhriciis lerrestris. (Oligoch.) Taken in Munich.

Monocystis catenata ( = partim. M. herculea Hesse 1909)

Muslow (1911) Arch. Protistenk., 22:51.

Spor. spher. 425/i, in chains. Cysts nearly spher. SOO^u. Spores 14X6;u.

Host: Lumbricus terrestris. (Oligoch.)

Monocystis minima

Konsulofif (1916) Arch. Protistenk., 36:353-61.

Spor. ovoidal, 42/^, spores ellipsoidal. In long. Intestinal par.

Hosts: Euchlanis dilatata. (Rotif.); Salpina mucronata Ehrbg. (Rotif.)

Taken at Sofia.

Monocystis perforans

Pinto (1918) Brazil-Medico; (1919) Contribuijao ao estudo das Gre-

garines, Rio de Janeiro, 113 pp., 6 pi.

Sem. ves. Spor. ovoidal to cylindr. in chains. 1200/iX800At. Nucl. ellip-

soidal, cysts spher. spores 24X7.5;u.

Host: Glossoscolex wiengreeni. (Ann.) Taken at Rio de Janeiro, Brazil.

142 MINNIE W. KAMM

Monocystis niichaelseni

Hesse (1916) Tolosani Monit. Zool. ital , 27:217-22.

Monocystis sp.

Wellmer (1911) Schr. Physik. Ges. Konigsb., 52:147. Coleomic.

Host: Helophorus aquaticus. (Col.) Taken in East Prussia.

Genus Lithocystis Giard 1876

Emend. Pixell-Goodrich (1915) Q. J. Mic. Sci., 61:81-104. Spor.

elongate, very motile. Spores in rosettes, long ovoidal, truncate. Epispore

a funnel at one end through which 8 sporozoites escape, other end a tubular

tail.

Lithocystis foliacea

Pixell-Goodrich (1915) Q. J. Mic. Sci., 61:81-104.

Coelomic. Max. 1. sporont 1.3 mm. Cysts spher. 600yu. Spores long-

ovoidal, 24X9/X, tail three times as long as spore, with leaf-like

expansion, funnel at other end.

Host: Echinocardium cordatum. (Echinodermata.) Taken at Naples and

Plymouth, Eng.

Lithocystis microspora

Pixell-Goodrich (1915) Q. J. Mic. Sci., 61:81-104.

Coelomic. Max. 1. 1mm. Cysts spher. 300^t. Spores 13X7/x, with tail

two or three times as long, narrow, tapering.

Host: Spatangus purpureus. (Echinod.) Taken off Plymouth.

Genus Urospora Schneider 1875

Emend. Pixell-Goodrich (1915) Q. J. Mic. Sci., 61:81-104. Tropho-

zoites coarsely granular, elongate. Female gamete with long flagellum,

male non-motile. Cysts spher. 8 sporozoites which escape from one end of

funnel-shaped epispore other end a filamentous tail.

Urospora synaptae { = Syncystis syn. Cuenot (1891) Rev. Biol. nord. Fr.,

3:295.)

Cuenot (1912) Bull. sta. biol. Arcachon, Bord., 14:85.

One form spor. rotund, 300;u max. diam, other vermiform SOO/jl long, very

motile. Cysts spher. 150/x, spores ovoidal, 20/x, one end cupped other a

long filament. Coelomic and intestinal.

Host: Synapta gaUicnnci. (Echinod.) Taken at Arcachon and Roscoff, Fr.

Urospora travisiae

Urospora ovalis

Mawrodiadi (1914) Varsava Univ. izv., No. 8, 1-164.

NEW GREGARINES DESCRIBED FROM 1911-1920 143

Urosporz neapolitana

Pixell-Goodrich (1915) Q. J. Mic. Sci., 61:81-104. See also Q. J. Mic.

Sci., 60:159-74.

Spor. 200-300^ 1., 40^ w. Cysts spher. 100-200^. Spores 12X7^, ovoidal,

cupped at one end, tail twenty times as long as spore and tightly coiled

at other end.

Host: Echinocardiuni cordatum. (Echinod.) Taken at Naples.

Urospora echinocardii

Pixell-Goodrich (1915) Q. J. Mic. Sci., 61:81-104.

Troph. and cysts same as U. neapolitana. Spores \9n long, tails 6 or 7

times as long as spore, not tightly coiled.

Hosts: Echinocardium sp. and Spatangus sp. (Echinod.) Taken at Ply-

mouth, Eng.

Genus Gonospora Schneider 1875. Emend

Pixell-Goodrich (1916) Q. J. Mic. Sci., 61:205-16. Polymorphic,

nematoid, pyriform or ovoidal. Cysts spher., spores with funnel at one

end refringent endo-spore which gives oflf processes supporting thick trans-

parent ectospore and funnel at other.

Gonospora mercieri { = Lithocystis miilleri Giard 1886.)

Cuenot (1912) Bull. sta. biol. Arcachon, Bord., 14:88-90.

Spor. spher., max. diam. 160/x. Cysts 180/i in diam., spores 23/z long,

ovoidal, no caudal filament. Intestinal par.

Kost: Synapta digiiaia. (Echinod.) Taken at Arcachon, France.

Gonospora glycerae

Pixell-Goodrich (1916) Q. J. Mic. Sci., 61:205-16.

Coelom par., generally surrounded by host epithelium. Spor. 1 to 5 mm.

long, widest near ant. end and tapering to blunt point. Nucl. spher. cysts

spher. spores 10X8^t. Refringent endospore with many supporting proc-

esses. Associations of spor. by 'ball-and-socket' dovetailing of ant. ends.

Host: Glycera siphonostoma. (Polych.) Taken at Naples.

Gonospora testiculi ( = Cysiobia test.)

Tregouboff (1916) C. R. Soc. Biol, 79:652-5; (1918) Arch. zool. exper.,

57:471-509.

L. 250//, elongate-ovoidal rounded at ant. end, pointed at post. end.

Cysts 60-IOOfjL in diam. Spores 8 to 10/x in 1. Testicle par.

Host: Cerithium vulgatum. (Moll.) Taken at Villefranche-sur-Mer,

France.

Gonospora intestinalis { = Cystobia int.)

Sokoloff (1914) Arch. Protistenk., 32:221-8.; Tregouboff, (1918) C. R.

Soc. Biol., 79:652-55, Pixell-Goodrich (1916) Q. J. Mic. Sci., 61-205-16.

144 MINNIE W. KAMM

Intestinal par. Spor. elongate, max. 1. 300/1. Cysts nearly sphcr. SOO^t.

Spores ovoidal, IO/jl 1.

Host: Glycera siphonstoma. (Polych.) Taken at Naples.

Genus Rhynchocystis Hesse 1909

Spor. ovoidal to cylindr. ant. end conical. Spores biconical with like

poles.

Rhynchyocystis hessei

Cognetti de Martiis (1911) Mem. R. Accad. Sci. Torino, 46:207-16.

^ax. 1. spor. 116/i, w. 88/x. Coelomic par. Spores 13X2.5/i.

Host: Pareudrilus palUdus. (Polych.) Taken in 'Equatorial Africa.'

Rhynchocystis geoplanae

Fuhrmann (1916) Centrallbl. Bakt. Parasit., 77:482-5.

Parenchymatous and intestinal par. Largest spor. 280X80/x. Nucl.

large, spher. Cysts spher. 180/^. 'Pseudoepimerite' a rosette.

Hosts: Geoplana backi, G. amagensis. (Furh.) Taken in Columbia, S. A.

Genus Diplocystis Kiinstler 1887

Coelomic, associating early to form spherical masses. Spores spherical

or oblong. Eight sporozoites.

Diplocystis phryganeae

Berg-von-Emme (1913) Arch. Protistenk., 28:43-51.

Spor. subspher. nucl. spher.

Host: Pkryganea grandis (Neur.) Taken at Petrograd.

Genus Lankesteria Mingazzini 1891

Trophozoites spatulate, cysts spher., spores ovoidal.

Lankesteria sp.

Swarczewsky (1910) Festschr. Geburtst. R. Hertwig 1: 635-74; (1911)

Arch. Protistenk., 22:236.

Intestinal par., encysted in parenchyme. Cysts spher. lOO/i. Cyst walls

dissolve and spores are carried to organs, set at liberty at death of host.

Hosts: Planaria sp. and Sorocoelis sp. (Plath.)

Lankesteria culicis

Stevenson and Wenyon Jour. Trop. Med. and Hyg., 18:196; Macfie

(1917) Report of the Accra Lab. for 1916, London, pp. 67-75.

Host: Stegomyia fasciata larv. (Dipt.) Taken at Accra, Gold Coast,

Africa.

. Genus Ancora Labbe 1899

Anchor-shaped spor. two long lateral backwardly-directed prolonga-

tions from ant. end, body tapering to sharp point.

Ancora lutzi {?A. sagittata Lcuckart Arch. Naturg., 26 (2):263)

Hasselmann (1918) Brazil-Medico, Aug. 10, 1918.

Host: Capitella capitata Fabr. (Ann.) Taken at Manguinhos, Rio de

Janeiro, Brazil.

new gregarines described from 1911-1920 145

Uncertain Genus in the Acephalinae

Genus RhytidocysHs Henneguy 1908

Trophozoite stage intracellular, encystment solitary, two sporozoites

in spore.

Rhytidocystis henneguyi

deBeauchamp (1912) C. R. Acad. Sci. Par., 154:1384; (1913) Arch.

Protistenk., 31:138.

Spor. ellipsoidal, encystment solitary. Nucl. spher. Spores 12X7yu,

ovoidal, symmetrical. Epithelium and lumen of intest.

Host: Ophelia negleda. (Polych.) Taken off Roscoff, France.

Uncertain Species in the Acephalinae

Unnamed sp.

Guenther (1914) Zool. Anz., 44:264-7.

Host: Ficalbia dofleini. larv. (Dipt.) Taken on Island of Ceylon. Habi-

tat: Tracheae and coelom.

Unnamed sp.

Pixell-Goodrich (1916) Q. J. Micr. Sci., 61:205-16.

Max. 1. 1.6 mm., w. 1 mm. Coelomic and attached to body or intest. walls.

Nucl. large, ovoidal.

Host: Glycera siphonostoma. (Echinod.) Taken at Naples.

Two other unnamed sp. found by same author, and in same host as last.

An Annotated List of the Species in the Suborder Schizogregarinae

Tribe 1. MONOSPORA Leger and Duboscq 1908

Family 1. OPHRYOCYSTIDAE Leger and Duboscq 1908

Tribe 2. POLYSPORA Leger and Duboscq 1908

Family 2. SCHIZOCYSTIDAE Leger and Duboscq 1908

Genus Schizocystis Leger 1900

Schizonts extracellular, vermiform, multinucleate. Gametes ovoidal,

pointed at one end. Cysts subspher. or ovoidal, spores octozoic, biconical.

Schizocystis spinigeri

Machado (1913) Mem. Inst. Oswaldo Cruz, Rio de Jan., 5:5-13.

Spor. slender, striated longitudinally, cysts ovoidal, spores ovoidal, pointed.

Both sporogony and schizogony noted.

Host: Spiniger sp. (Hemipt.) Taken near Manguinhos, Rio de Janeiro,

Brazil.

Family 3. SELENIDIIDAE Brazil 1907

Schizonts intracellular, multinucleate, at close of development. Game-

tocystes mobile, longitudinal myonemes. Parasitic in Polychaetes.

146 MINNIE W. KAMM

Genus Sclenidium Giard 1884

Schizogony in the intracellular stage.

Selenidium cruzi

Faria, Cunha and Fonseca (1917) Brazil-Medico, 31:243; (1918) Mem.

Osw. Cruz, 10:17.

Largest trophoz. 160mX25;u, vermiform, slightly flattened, ant. end blunt

with small pointed epimerite. Nucl. ellipsoidal.

Host: Polydora socialis. (Polych.) Taken at Rio de Janeiro, Brazil.

Selenidium mechnikovi

Leger and Duboscq (1917) Ann. Inst. Past., 31:69.

Intestinal par. Intra- and extra-cellular, schizozoites pyriform, 5/i.

Sporonts cucumber-shaped, 30-34^t, longit. striated, nucl. sub-spher.

Host: Glossobalanus minutus. (Enteropneusta.) Taken at Sainte-Jean-

de-Luz, France.

Family 4. MEROGREGARINIDAE Porter 1908

Family 5. SPIROCYSTIDAE Leger and Duboscq 1915

Arch. Protistenk., 35:199-211. Mono-sporic and monozoic, schizogony

and sporogony in same host.

Genus Spirocystis Leger and Duboscq

1911 Bull. Soc. zool. Fr., June, 1911.

Spirocystis nidula. Type species

Leger and Duboscq (1911) Bull. Soc. zool. Fr., June 1911; C. R. Soc.

Biol., 76:296; Arch. Protistenk., 35:199.

Sporocyst ovoidal, 35n long, rejected with excrement. Releases in its next

host through a micropyle a single folded sporozoite 40^i long. Sporozoite

gives rise to helix-shaped schizont found in somatic or visceralperitoneum.

This becomes multinucleate of max. 1. 35ju and gives rise to macro- and

micro-gametes, the copulation of which produces the spore, found in the

chloragogue cells.

Host: Lumbricus variegatus. (Oligoch.) Taken near Grenoble, France.

Tribe 3. OCTOSPOREA Keilin (1914) C. R. Soc. Biol., 76:768

With eight spores in the sporogonic cycle.

Family 6. CAULLERYELLIDAE Keilin (1914) C. R. Soc. Biol., 76:768.

Genus Catdleryella Keilin (1914) C. R. Soc. Biol., 76:768

Intestinal par. Schizogony extracellular, veg. nucleus gives rise to 16

merozoites. Each of the two sporonts in a cyst produces 8 gametes. These

16 conjugate by twos to form 8 spores which produce 8 sporozoites.

Caulleryclla aphiochaetae. Type species

Keilin (1914) C. R. Soc. Biol., 76:768.

Veg. stage 22^ long, ovoidal, pointed at end, embedded in epithelium.

NEW GREGARINES DESCRIBED FROM 1911-1920 147

Nucl. divides four times, giving rise to 16 merozoites liberated and affix

themselves to epithelium. Sporulation by twos with 16 gametes produced.

Cysts and gametes spherical.

Host: Aphiochaeta rnfipes larv. (Dipt.) Taken at Paris.

Caulleryella anophelis

Hesse (19?) C. R. Acad. Sci. Par., 166:569.

Spor. 35X30^, syzygy in twos, cysts spher. 24/x to 3>2ix. Spores sub-spher.

12.5X11m- Dehiscence of cyst in host intest.

Host: Anopheles bifurcatus. larv. (Dipt.) Taken in the Dauphine,

France.

Family of Uncertain Position

Family 7. POROSPORIDAE Leger and Duboscq 1908

1915 C. R. Soc. Biol., 75:95-8. Many sporozoites from a sporoblast.

No sporocyst.

Genus Porospora Schneider 1875

Epim. minute, button-like, spor. septate, usually solitary.

Porospora legeri

deBeauchamp (1910) C. R. Acad. Sci. Par., 151:997-9.

Spor. associative prot. of primite depressed at apex, satellite longer with no

prot. 750/iX75iU. Cysts spher. from two sporonts. Intestinal par.

Host: Eriphia spinijrons. (Crust.) Taken at Saint- Jean-de-Luz, Fr.

(This species was omitted from Sokolow's List-1911.)

Porospora portunidarum Leger and Duboscq 1911 ( = Aggregata p. Frenzel)

(1911) Arch. zool. exper., (5) 6:lix-lxx; (1913) C. R. Acad. Sci. Par.,

156:1932; (1913) C. R. Soc. Biol., 75:95.

Porospora pisae

Leger and Duboscq (1911) Ann. Univ. Grenoble, 23:403; Tregouboff

(1916) Arch. zool. exper., 55:xxxv-xlvii.

1 mm. long, eel-shaped. Encystment from one or two spor.

Host: Pisa gibosii. (Crust.) Taken at Cette and Villefranche-sur-AIer, Fr.

Porospora maraisi

Leger and Duboscq (1912) Ann. Univ. Grenoble, 23:399.

Host: Portunns depuraior. (Crust.)

Porospora nephropsis

Leger and Duboscq (1915) C. R. Soc. Biol, 75:368-71.

Spor. elongate, ellipsoidal, blunt at ends, max. 1. 240;Lf, max. w. 44/i. Nucl.

spher. Solitary vermiform enigmatic individuals 1300X36 also present.

Cysts 160/i in diam. Schizogonic spores 5 in diam.

Host: Nephrops norvegicus. (Crust.)

148 MINNIE W. KAMM

The classification of this family is uncertain because the sporonts are

apparently typical cephaline Eugregarinae yet a schizogonic cycle exists.

Minchin (1912) says 'The classification of the future will probably be one

which divides all gregarines into Cephalina and Acephalina and distributes

the schizog^egarines amongst these two divisions.'

Genus of Uncertain Position

Genus Selysina Duboscq 1917 C. R. Acad. Sci., 164:?

Selysina perforans. Type species

Duboscq (1917) C. R. Acad. Sci., 164:? (1918) Arch. zool. exper.,

?:l-53.

Host: Stolonica socialis. (Ascid.) Taken off Roscoff, France.

Unnamed gregarine

Strickland (1913) Jour. Morphol., 24:84.

Schizogonous. Pathogenic effect upon host. Inhabits various tissues.

Cysts spher. 250^t.

Host: Simulium bracteatum larv. (Dipt.) Taken near Boston, Mass.

A species described as Microtaeniclla clymenellae n.g., n. sp. from

Clymenella torqtiata (Ann.) by Calkins (1915) Biol. Bull., 29:46 is regarded

by the author as a colonial gregarine resembling the scolex and proglottids

of the cestodes, each segment being nucleated. This polynucleate condition

makes its inclusion in this group doubtful. Poche (Arch. Protistenk.,

37:6) considers it identical with the genus Haplozoon.

List of Hosts With Their Gregarine Parasites

Host Parasite

Platyhelminthes

Geo plana hacki Rhynchocystis geoplanae Fuhrman

G. amagensis Rhynchocyslis geoplanae

Planaria sp. Lankcstcria sp. Swarczewsky

Polyporus sulphureus Gregarine form, Wellmer

Sorocoelis sp. Lankestcria sp. Swarczewsky

Annelida: Polychaeta

Capitella capHala Aticora lutzi Hasselmann

Clymenella torquata Microtaeniella clymenellae Calkins

Glycera siphotrostoma Gonospora glyccrae Pi.xell-Goodrich

Glycera siphonosloma Gonospora inlcstinalis Pixcll-Goodrich

Glycera siphonosloma Three unnamed parasites Pixell-Goodrich

Ophelia neglecla Rhytidocyslis henneguyi dcBeauchamp

Parendrilus pallidus Monocyslis pareiidrili Cognetti de Martiis

Pareiidrilus pallidus Rhynchocyslis hessci Cognelti de Martiis

Polydora ciliala Polyrhabdina polydorae Caullerj'^ and Mesnil

Polydora socialis DoUocyslis sp. Faria, Cunha and Fonseca

Polydora socialis Selcnidium cruzi Faria, Cunha and Fonseca

Pygospionis scticornis Polyrhabdina pygospionis Caullcr>' and Mesnil

i

NEW GREGARINES DESCRIBED FROM 1911-1920

149

Host

Rhinodrilus inccrlus

Scolclepsis fuliginosa

Spio marlinensis

ANNELroA: Oligochaeta

Kynotus Pittarcllii

Lumbricus terrestris

Lnmhricus terrestris

Lumbricus variegatus

Glossoscolex mengreeni

Annelida: Hirudinea

Glossophonia complanata

Hemiclepsis marginala

ROTIFERA

Euchlanis dilatata

Salpina mucronata

ECHINODERMATA

Eckinocardium cordatum

Echinocardium cordatum

Echinocardium sp.

Spatangus sp.

Synapla pitrpurcus

Synapta galliennei

Synapta digitata

MOLLUSCA

Cerithium vulgalum

Crustacea

Ampelisca spinipes

Anaspides tasmaniae

Atyaephyra Dcsmaresti

Balanus amphilrite

Balanus eburneus

Eriphia spinifrons

Gammarus marimis

Libinia dubia

Nephrops norvegicus

Portumis depurator

Pisa gibosii

Talitrus saltator

Talorchestia longicornis

Uca pugnax

Uca pugilator

Chilopoda

Scolopendra heros

Scolopendra subspinipes

Scolopeiidra sp.

Scolopendra sp.

Scolopendrella sp.

Parasite

Monocystis Ihamnodrili Cogn. dc Marliis

Polyrhabdina spiofiis Caullery and Mesnil

Polyrhabdina brasili Caullery and Mcsnil

Taeniocystis legcri Cogn. de Martiis

Monocystis roslrata AIuslow

Monocystis catenala Muslow

Spirocystis nidula Leger and Duboscq

Monocystis perforans Pinto

Metamera schubergi Duke

Monocystis minima Konsuloff

Monocystis minima Konsulofif

Lithocystis foliacea Pixell-Goodrich

Urospora neapolitana Pixell-Goodrich

Urospora echinocardii Pixell-Goodrich

Lithocystis microspora Pixell-Goodrich

Urospora synaptae Cu6not

Gonospora mercieri Cuenot

Gonospora tesliculi Tregouboff

Ceplmloidoplwra ampelisca Kamm

Ganymedes anaspides Huxley

Uradiophora cuenoti Mercier

Pyxinoides balani Tregouboff

Unnamed parasite, Buddington

Porospora legeri deBeauchamp

Cephaloidophora macidata Leger and Duboscq

Cephaloidophora olivia Kamm

Porospora nephropsis Leger and Duboscq

Porospora maraisi Leger and Duboscq

Porospora pisae Leger and Duboscq

Cephaloidophora talilri Mercier

Cephaloidophora delphinia Kamm

Cephaloidophora nigrofusca Kamm

I

Amphorocephalus amphorellus Ellis

Nina indicia Merton

Echinomera magalhaesil Kamm

Seticephalus elegans Kamm

Gregarina brasilietisis Pinto

Gregarine form, WeUmer

150

MINNIE W. KAMM

Host

DiPLOPODA

Callipits laclarius

Euryiirus crythropygiis

Fontaneria coarctata

Orthomorpha coarctata

Orthomorpha gracilis

Orthomorpha sp.

Parajulns impressus

Parajulus venustus

Parajulns sp.

Rhinocricus pugio

Rhinocricus sp.

Thysanura

Sminthurus jiiscus

Orthoptera

Ceuthophilus latcns

Ceuthophilus maculatus

Ceuthophilus ncglcctus

Ceuthophilus stygius

Ceuthophilus valgus

Conocephalus f rater

Encoptolophus sordidus

Forficularia auricular ia

Gryllus ahbrcviatus

Ischnoptera pennsylvauicus

Mclanoplus differential is

M elano plus femur-ruhr urn

Udeopsyllae nigra

Hemiptera

Spiniger sp.

Neuroptera

Aeschnidae Iv.

Acschna sp.

Phryganea grandis

Sympelrum rubicundulum

Tramea lacerata

DiPTERA

Anopheles bijurcalus Iv. ,

Aphiochacta rufipes Iv.

Ceratophyll us fasciatus

CcratophylUis farreni

Ccratophyllus Jringillac Iv.

Ccralophyllus gallinae Iv.

Ceralophyllus gallinae ad.

Ccratophyllus slyx

Stenophora

Fonsecaia j.

Stenophora

Parasite

lactaria Watson

diplocorpa Watson

Cauda ta Watson

clongata Ellis

robusta Ellis

wlymorpha Pinto

imprcssa Watson

robusta Ellis

cockcrellae Ellis

cunhai Pinto

lutzi Pinto

cruzi Pinto

viannai Pinto

umbilicata Pinto

tenuicoUis Pinto

Gregarine form, Wellmer

Grcgarina

Gregarine

Grcgarina

longiducta Ellis

neglecta Watson

stygia Watson

consobrina Ellis

chagasi Pinto

nigra Watson

form, Pantel

galliveri Watson

illinensis Watson

nigra Watson

udeopsyllae Watson

Schizocystis spiniger Machado

Bolhriopsis claviformis Pinto

Aclinocephalus brachydactylus Ellis

Diplocyslis phrygancac Berg-von-Emme

Prismatospora cvansi Ellis

Caullcryclla anophelis Hesse

Caulleryclla aphiochaelae Keilin

A grip pi na bona Strickland

Steinina rotundata Ashworth and Rettie

Aclinocephalus parvus Wellmer

Stein itui rotundata Ashworth and Reitie

Steinina rotundata Ashworth and Rettie

NEW GREGARIXES DESCRIBED FROM 1911-1920

151

Host

Ficalbia dofleini Iv.

Simidiiitn hractealum Iv.

Stegomyia fasciaca Iv.

COLEOPTERA

Alobaks poinsyliHuticus

A viara anguslota

Asida opaca

Asida sp.

Broscus ccphaloks

Carabus sp.

Clerid Iv,

Coccinella sp.

Coccinella novcnniolala

Coptotomus inkrrogatus

Coploiomus intcrrogalus

Crypticiis quisquilius

Ciicujus Iv.

Cyclirus roslralus

Dermesles lardariiis

Diabrotica vitkila

Elakridae Iv.

Ekodes sp.

Eusattus sp.

Harpalus aeneus

Har pains pennsylvankus

Harpalus pennsylvanicus erythropiis

Harpalus pennsylvankus longior

Harpalus ruficornis

Heledona agricola

Helophorus aquaticus

Hydrophilus akrrimus Iv.

Hydrophilus sp.

Hylobius abktis

Ips typographus

Lagria hirla

Lepkchirus edax

Leptochirus edax

Ninus inkrstitialis

N ycktheres barbaraia

Omoplaia normalis

Platydema excavatum

Platynusruficollis

Procrusks coriaceus

Pkrostkhus niger

PkrosHchus niger

Syskna sp.

PkrosHchus slygkus

PkrosHchus stygicus

Pkrostkhus vulgaris

Par.\site

Unnamed par. Guenther

Unnamed par. Strickland

Lankeskria cidicis Stevenson and Wenj'on

Actinocephalus zophus Ellis

Skinina rotunda Watson

Stylocephalus giganteus Ellis

Stylocephulus giganteus Ellis

Gregarina erecta Wellmer

Cometoides sp. Wellmer

Bulbocephalus wardi Watson

Gregarina Jragilis Watson

Gregarina katherina Watson

Gregarina globosa Watson

Gregarina coptolomi Watson

Gregarina ovoidea Wellmer

Bulbocephalus elongatus Watson

Gregarine form, Wellmer

Pyxinia bulbifera Watson

Gregarina diabrotica Kamm

Gregarina gracilis Watson

Stylocephalus giganteus Ellis

Gregarina polyaulia Wellmer

Actinocephalus gimbeli Watson

Hirmocustis harpali Watson

Steinina harpali Watson

Gregarina polyaulia Wellmer

Gregarine form, Wellmer

Monocystis sp. Wellmer

Cometoides-like form, Wellmer

Bothriopsis tcrpsichorella Ellis

Gregarina hylobii Kamm

Gregarina typographi Fuchs

Gregarina rostrata Wellmer

Actinocephalus crassus Ellis

Stylocystis ensiferus Ellis

Gregarina guatemalensis Ellis

Actinocephalus zophus Ellis

Gregarina watsoni Pinto

Gregarina platydema Kamm

Gregarina platyni Watson

Actinocephalus permagnus Wellmer

Gregarina exiguus Kamm

Actinocephalus echinatus W^ellmer

Gregarina aragaoi Pinto

Gregarina monarchia Watson

Gregarina intestinalis Watson

Actinocephalus echinatus Wellmer

152

MINNIE W. KAMM

Host

Tenehrio castaneus

Tenebrionidae Iv.

Tribolium ferrtigineum

Tribolium ferrugineum

Tribolium ferrugitieum

Tribolium ferrugineum

Tritoma quadripustulata

Lepidoptera

Endrosis fenestrella Iv.

Oecophora pseudospretella Stain

Tinea pallescentella Stain

Arachnida

Ctenocephalus scrraticeps

Oribata geniculala

TUNICATA

Stolonica socialis

Enteropneusta

Glossobalanus minutus

Parasite

Gregarina grisea Ellis

Grcgarina tenebrionella Watson

Gregarina minuta Ishii

Gregarina crassa Watson

Disymophyes minuta Kamm

Steinina obconica Jshii

Gregarine form, Wellmer

Leidyana tinei Keilin

Unnamed greg.

Grcgarina ctenocaphalus Ross

Gregarina sp. Wellmer

Selysina perforans Duboscq

Selenidium metchnikotd Leger and Duboscq;

DEPARTMENT OF METHODS, REVIEWS, ABSTRACTS, AND

BRIEFER ARTICLES

ABNORMAL EARTHWORM SPECIMENS, HELODRILUS SUBRU-

BICUNDUS AND H. TENUIS*

By

Frank Smith

University of Illinois

Comparatively little attention has thus far been given to abnormalities

in the relations of the reproductive organs of earthworms. Variations from

the normal positions and number are sometimes found, and asymmetri-

cally placed gonads and openings of efferent ducts are not infrequently

encountered. Since further investigation of such abnormalities may lead

to at least a partial understanding of their relation to disturbances in the

normal developmental activities of the animals concerned, it seems advisable

to record the more important details in the structure of specimens repre-

sentative of some of the more common types of such abnormalities, if

indeed it be found that there are such types.

A specimen of Hclodrilus siibriibicimdus (Eisen) recently collected at

Urbana, Illinois, in the banks of a stream heavily contaminated with

sewage was found to have the spermiducal pores on somite 14 instead of in

the usual position on the fifteenth somite. Sagittal sections of the left half

of the anterior part were made and unexpected irregularities were found.

Spermaries and spermiducal funnels are present in the usual positions in 10

and 11. An ovary and oviducal funnel are present in the usual positions in

13; but an additional one of each, equally well developed, have similar

positions in the twelfth somite which normally has no gonads. An oviducal

pore is present in the usual position on 14, and in addition there is a super-

numerary one on 13, related to the oviducal funnel of 12. The spermiducal

pores on 14 are slightly laterad of the oviducal pores of the same somite.

Sperm sacs in 9, 11, 12, and an ovisac in 14 have the usual location and

relations. The calciferous gland, crop, and gizzard also have the usual

location and relations; but the most posterior heart is in 10 instead of in

the usual position in 11; and the lateral longitudinal vessel branches off

from the dorsal vessel in 11 instead of in the usual place in 12. The ventral

setae of 9 are modified to genital setae which are of about twice the length

of ordinary setae and relatively more slender.

*Contribution from the Zoological Laboratory of the University of Illinois, No. 205.

153

154 FRANK SMITH

The presence of extra gonads in 12 is not infrequently met with, but the

presence of spermiducal and oviducal pores on the same somite (14) is

decidedly unusual, in the experience of the writer, but has been found also

in another specimen, described below.

A specimen of Helodrilus tenuis (Eisen) collected near Urbana, Illinois

in a fallen and decaying tree, attracted attention because the spermiducal

pores were asymmetrically placed, the one on the right side being normally

situated on 15, while that of the other side opened on the somite next ante-

rior. Sections were made and the asymmetrical relations were found to

extend to internal organs. Reproductive organs of the right side were

found in normal positions and relations, as follows: spermaries and spermi-

ducal funnels in 10 and 11; an ovary and oviducal funnel in 13; oviducal

pore on 14; and the spermiducal pore on 15. In the left half of the worm,

there are spermaries and spermiducal funnels in 9, 10, 11; ovaries and

oviducal funnels in 12 and 13; oviducal pores on 13 and 14; and a spermidu-

cal pore on 14, laterad of the oviducal pore of that somite. The extra

gonads and associated funnels are as large and well developed as the normal

ones. Paired sperm sacs in 11 and 12 are in the locations normal for this

species. No irregularities in the location of hearts and lateral longitudinal

vessels have been noticed; and the alimentary tract has normal relations,

except that the anterior evagination of the calciferous gland in the left

half of the worm is found anterior to the septum 9/10, and the one in the

right half is anterior to 10/11 which is the more normal position.

Asymmetry in the number and position of various organs in the right

and left halves of specimens is of fairly frequent occurrence and often

involves circulatory and alimentary systems as well as the reproductive

organs. It will be noticed that the presence of both spermiducal and ovi-

ducal spores on 14 is associated, in the two specimens described above, with

the presence of ovaries and oviducal funnels in both 12 and 13; but such

association may be a mere coincidence rather than an actual correlation.

SUBSTITUTES FOR ABSOLUTE ETHYL ALCOHOL

By

Lawrence E. Griffin

Reed College

During the past year the writer has had his interest attracted to the

question of whether other alcohols can be successfully substituted for

anhydrous ("absolute'') ethyl alcohol in histological work. For a consider-

able part of the year the air of Portland, Oregon, is nearly saturated with

water vapor, and the tendency of absolute ethyl alcohol to absorb water

constitutes one of the difficulties in its use. More potent reasons for seeking

substitutes were, however, the high cost of the absolute ethyl alcohol and

annoyances resulting from tax and prohibition laws. Even when alcohol

can be secured by institutions free of tax there are regulations to be ob-

served which entail a certain amount of delay in securing it, as well as much

supervision of its use. So it would be advantageous if other alcohols can

be used which are not subject to the regulations of the Bureau of Internal

Revenue, and which are not sought as beverages. Fortunately, there are

at least three such alcohols which have proved to possess merit.

METHYL ALCOHOL. The ordinary commercial quality of methyl

alcohol contains about 95% of alcohol, the remainder consisting mostly of

acetone, with traces of a large variety of other impurities. Purified methyl

alcohol, anhydrous, and nearly free from acetone and other impurities, is

put on the market under various trade names. The brand which we have

tested is known as Diamond Methyl alcohol. Being practically free from

acetone it not only lacks the strong disagreeable smell of ordinary wood

alcohol but, in fact, has a pleasant odor much like that of refined grain

alcohol. This alcohol dehydrates sections and tissues as well as the absolute

ethyl alcohol, and is a little more reliable because it will dehydrate more

sections than an equal amount of absolute grain alcohol. We have found it

to be a good solvent, and have used it with success in the compounding of a

number of reagents. As regards cost, it was not only far cheaper than anhy-

drous ethyl alcohol, but was considerably cheaper than the tax-free 95%

grain alcohol which we bought a short time before we secured the methyl

alcohol. We are now using this alcohol as our standard reagent in dehydra-

tion. In the course of eight months our stock, kept in a large glass stop-

pered bottle, has not absorbed enough water vapor to be noticeable. We

purposely made no particular effort to seal the stock bottles tightly from the

atmosphere. Anhydrous ethyl alcohol kept under the same conditions

would have been useless for the dehydration of tissues.

155

156 LAWRENCE E. GRIFFIN

BUTYL ALCOHOL. Professor George W. Martin has called attention

(Science, April 21, 1922), to the use of butyl alcohol in dehydration and

infiltration with parafin. This alcohol has been under test in our laboratory

as a dehydrating reagent for several months and has given excellent results.

We have used it, however, only in the last stage of dehydration, passing

slides from 90% methyl or 95% ethyl alcohols to the butyl alcohol. It

appears to us to be superior to either ethyl or methyl alcohols for dehydra-

tion, but its use is slightly disagreeable on account of the pungent, char-

acteristic odor. Inhalation of its fumes causes a slight, temporary irritation

of the throat. In reply to our inquiry as to whether this property of butyl

alcohol might be removed, the Commerical Solvents Corporation, which

made our sample, replied:

"The irritation of the throat, caused by the use of Butanol, is quite

characteristic of this compound and is a property which it would be difficult

to obliterate. However, we do not believe it has any harmful effect, as

some of the men who work on the distillation end of the process have been

subjected to this for years and have experienced no ill effects. They are

much less susceptible to the irritation after having worked with this com-

pound for some time."

Butyl alcohol is, at any rate, a valuable reagent for dehydration, our

observation being that the sections dehydrated with it are slightly more

brilliant than those cleared with the previously mentioned alcohols. As

Professor Martin also states, butyl alcohol is a solvent of paraffin, and can

be used for infiltration of tissues likely to shrink or harden in the usual

infiltrants. We believe, however, that for this purpose it is surpassed by

Terpeneol.

TERPENEOL. The use of terpeneol (terpineol) in place of absolute

ethyl alcohol was suggested a number of years ago, but it is only lately that

I have been able to test it thoroughly. Terpeneol is a pleasant smelling,

aromatic liquid, of about the consistency of thin cedar oil. It is tolerant of

large amounts of water in dehydration, and also dissolves paraffin and

resins. On account of its consistency we have found it advisable to use

first a mixture of terpeneol and methyl alcohol before placing tissues into

pure terpeneol. Terpeneol may be used as a dehydrating agent for sections,

but does not have any advantages over methyl or butyl alcohol when used in

that way. As it dissolves parafin readily it is more useful as a dehydrant

and infiltrant of tissues to be embedded. Terpeneol dissolves parafin better

than butyl alcohol. Our experience has been that tissues which had been

dehydrated and infiltrated with terpeneol were less shrunk and hardened

than when embedded by the ordinary methods. Terpeneol is of rather high

refractive index, so that it serves as a clearing agent also. Sections may be

transferred directly from terpeneol to Xylol-damar. As the terpeneol

SUBSTITUTES FOR ETHYL ALCOHOL 157

does not make tissues so brittle as does Xylol it can be used advantageously

in the preparation of whole mounts.

We have also found that damar dissolved in terpeneol makes a mounting

medium which, on account of the refractive index being lower than that of

xylol-damar or xylol-balsam, shows some details of cell structure which

are obscured in these commonly used mounting media.

/yt

I

TRANSACTIONS

OF THE

American

Microscopical Society

Organized 187S Incorporated 1891

PUBLISHED QUARTERLY

BY THE SOCIETY

EDITED BY THE SECRETARY

PAUL S. WELCH

ANN ARBOR, MICHIGAN

VOLUME XLI

Number Four

Entered as Second-class Matter August 1.?, 1918, at the Post-office at Menasha.

Wisconsin, under Act of March 3, 1879. Acceptance for mailing at the

special rate of postage provided for in Section 1 103, of the

Act of October 3, 1917. authorized Oct. 21, 1918

iSltr dollrgiate Prraa

Geopoe Banta Publishing Compan-v

Menasha. Wisconsin

1922

TABLE OF CONTENTS

For Volume XLI, Number 4, October, 1922

The Anatomy of Some Sexually Mature Specimens of Dero limosa Leidy, with one

plate, by R. L. Mayhew 159

Studies on American Naid Oligochaetes, by H. E. Hayden, Jr 167

Excessive Sexual Development in Hydra oligactis with Spermary on Tentacle, with one

figure, by A. W. Schmidt 172

Some Suggestions for Teaching Mycology, with two figures, by F. D. Heald 175

List of Members 179

Index to Volume XLI 189

TRANSACTIONS

OF

American Microscopical Society

(Published in Quarterly Instalments)

Vol. XLI OCTOBER, 1922 No. 4

THE ANATOMY OF SOME SEXUALLY MATURE SPECIMENS

OF DERO LIMOSA LEIDY.^

By

Roy L. Mayhew

University of Illinois

Introduction

Ecology. The material on which this paper is based was collected

during the summer of 1921 at the Biological Station of the University of

Michigan, on Douglas Lake, Michigan. The particular locality was a

small bog draining into Burt Lake, about four and one half miles from the

Station. A thick mat of algae taken from a plank at the surface of the

water proved to be especially rich in the following genera of Naididae:

Nais, Chaetogaster, Pristina, and Dero. The families Lumbriculidae and

Enchytraeidae were represented by a number of specimens. Plankton

taken from the water near by contained specimens of the genera Slavina,

Stylaria, Nais, Dero, and Pristina.

During the examination of the algae it was frequently noticed that a

number of worms of the same species would be found close together.

Notes made at the time showed that four sexually mature specimens of

Dero limosa were thus found in close proximity. At other times Pristina

would be represented in great abundance. Five specimens belonging to

the genus Nais were taken from an area about two inches square and were

the only specimens of the genus found in the algae. Only one or two

specimens belonging to the genus Nais were taken from the plankton.

So commonly was the grouping of worms of the same kind observed, that

when one of a particularly desirable type was located, the algae of the

immediate vicinity was examined with special care.

Technique. Specimens of Dero were found to be the most difficult of

any of the worms encountered, to kill and fix for histological study.

Anesthesis with chloretone often resulted in the death and maceration of

1 Contribution from the Zoological Laboratory of the University of Illinois, No. 212

and from the University of Michigan Biological Station.

159

160 ROY L. MAYHEW

the posterior part of the worm before the anterior ceased to move or came

sufficiently under the influence of the drug to prevent bending and distor-

tion when subjected to the killing fluid. A concentrated aqueous solution

of corrosive sublimate was used as a fixing fluid. The specimens sectioned

were cut transversely, 6 ^ in thickness, and stained with Delafield's haema-

toxlin and eosin. Mounts, in toto, were stained in borax carmine and

cleared in oil of wintergreen.

Identification. Seven sexually mature and numerous immature speci-

mens of Dero limosa Leidy were taken from the algae referred to above

during the first two weeks of August, 1921. The determination of the

species proved a little difficult since living specimens not anesthetized

were constantly in motion, and, when anesthetized or kijled for preserva-

tion, were usually found to have completely closed the branchial pavilion.

The gill arrangement seems, however, identical with that of the above

named species as described by Michaelsen ('00, '03, & '09) and others.

Examination of sections of the closed pavilion reveals the fact that the

lateral portions of the lip, which are carried medially in the process of

closing, may easily be mistaken for a third pair of gills on the ventral

portion of the structure, since they are intimately associated with the

true gills in position and have similar cell arrangement (fig. 16).

Since the only descriptions found of sexually mature specimens be-

longing to the genus are those given by Michaelsen ('00) for D. perrieri

and D. imiltibranchiata, and by Walton ('06) in his general description of

the genus, it seems desirable to give a rather extended account of the new

material mentioned above. The descriptions of the sex organs as given

by Michaelsen are brief and are here quoted for comparison.

D. perrieri, "1 Paar Samentrichter im 5. Segm.; Samenleiter allmahlich

in die driisenlosen Atrien iibergehend. 1 unpaariger Eiersack. Samentas-

chen im 5. Segm., mit scharf abgesetztem, ziemlich kurzem, tonnenformi-

gem Ausfiihrungsgang."

D. multibranchiata, "Unpaariger Samensack vom 5. Segm. nach hinten

gehend. Ovarien (Eiersacke?) im 7. Segm."

Length and Number of Somites

The length of preserved sexually mature specimens varied from 5 to

10 mm. and the total number of segments from 40 to 55. Immature

specimens showed an equal range of variation in length and contained

from 40 to 62 somites; those with one or two budding zones containing

from 50 to 59.

Setae

In respect to the setae, the sexually mature specimens differ from the

immature only in the entire absence of ventral bundles in the 6th somite

\

ANATOMY OF DERO LIMOSA LEIDY 161

in contrast with the occurence of special or genital setae in Nais obtusa

and a number of other species described by Piguet ('06 & '09). The

dorsal bundles, which begin on the 6th somite in both mature and immature

specimens, have been found to contain one long capilliform and one

short, slightly cleft, biuncinate seta in each of the bundles examined

(fig. 13). The long capilliform setae are about equal in length to the

diameter of the body, and are without serrations. The biuncinate setae

extend but little beyond the surface of the body, and are more or less

curved and somewhat tapered distally. Their distal portions are much

stouter than figured by Michaelsen ('09) and by Bousfield ('87), and the

tips 6f the teeth are not sharp pointed but slightly rounded. One biunci-

nate seta of a sexually mature specimen was found to be somewhat smaller

in diameter and to have a slight nodulus. Piguet ('09) says that mature

specimens of the genus A^ais lose the setae of the dorsal bundles in the

clitellar somites, but such is not the case in D. limosa.

The ventral bundles (fig. 9-12) contain from 2 to 5, in the majority of

instances 3 or 4, biuncinate setae. The distal tooth, or the one farthest

from the nodulus, is about equal in length to the proximal, or but slightly

longer. The teeth are usually sharp pointed, though occasionally the

proximal one is thickened and round pointed. They often appear to have

round tips due to their being turned so that an exact lateral view is not

obtained. Each of the ventral setae is provided with a well defined nodu-

lus.

The ventral setae of somites 2-5 are longer than those from the re-

mainder of the body, in both mature and immature specimens. Setae

from each of the somites 2-5 were measured and averaged for each speci-

men with the result that the averages varied from 120/i to 160/x. Averages

obtained in like manner from a number of somites posterior to 6 varied

from 80 to 100;u. The extreme difference between the averages so obtained

for any specimen was 65 /x, the measurements being 95^1 to 160/u. Definite

ratio relations have been noted between the portions of the setae proximal

and distal to the noduli in particular parts of the worm. The portions

of the setae distal to the noduli in somites 2-5 are usually about equal in

length to the proximal parts but may be as much as 1.5 times as long, while

the distal portions of setae in somites posterior to 6 are only .6 to .8 as

long as the proximal.

The. Reproductive Organs

Clitellum. This organ extends from the anterior part of the 5th

somite almost to the setae of the 8th (fig. 1, cL). On the ventral side it

begins just back of the openings of the spermathecae, but dorsally farther

forward. In life the clitellum is yellowish or cream colored, and, when

sectioned, is found to be made up of a single layer of large columnar cells

162 ROY L. MAYHEW

containing large globules of a substance, which is probably the secretion

that later forms the cocoon, and relatively small nuclei irregularly dis-

tributed in the cells.

Spermathecae. These are conspicuous paired organs almost filling the

ventral two thirds of the 5th somite (fig. 1-3, spm.) The upper part is

a relatively thick walled sac with deeply staining, irregularly arranged

nuclei, and well filled with a compact mass of sperm cells indicating that

copulation has taken place. The duct arises a little laterad of the most

ventral portion of the sac and extends directly to the body wall where it

turns directly posteriad, a very short distance, and opens on the surface

through the pore slightly anteriad and laterad of the ventral seta bundle

of the same side of the somite (fig. 2 & 3). In one specimen the duct

passes almost directly to the pore. The walls of the duct are thickly set

with nuclei of about the same size as those of the sac. Dorsal to the

spermathecae there is a quantity of sperm cells in the body cavity.

Sperm ducts. A pair of sperm ducts lie in the 6th somite and have

their funnels in the posterior part of the 5th. Each of the pair consists

of four portions, (1) the spermiducal funnel, (2) a duct joining the funnel

to the atrium, (3) the atrium, and (4) the duct connecting the atrium with

the hypodermal invagination. The spermiducal funnels are found in the

ventral posterior part of the 5th somite anterior to septum 5/6. They are

shaped as if the distal end of the duct had been split on the dorsal side

and the lateral portions flattened out in the process of formation. The

part of the duct between the funnel and the atrium tapers posteriorly,

bends abruptly upon itself (figs. 1 & 7), and extends anteriorly to its point

of union with the atrium, on the median anterior ventral surface of the

latter. The atrium is a cylindrical sac occupying much of the anterior

ventral portion of the corresponding half of the 6th somite. The walls

are relatively thick and are made up of a single layer of large irregular

cells containing large vacuoles. The duct leading from its posterior end

is short and opens into a conspicuous invagination of the hypodermis on

the ventral wall of the body. In one specimen one atrium was displaced

so that it lay dorsad of the nerve cord in the median line of the body.

Spcrmaries. Nothing was found which could be identified as such.

This fact is probably due to the advanced stage of sexual maturity of the

specimens, as the sperm sac and spermathecae were filled with sperm

cells. When developed, they should be present on the posterior side of

septum 4/5, since sperm cells were found above the spermathecae, and

the sperm sac extended posteriad from the 5th somite.

Sperm sac. The sperm sac is a posterior evagination of septum 5/6,

and extends a short distance posteriad of the setae of the 7th somite in

one specimen.

ANATOMY OF DERO LIMOSA LEIDY

163

Oviducal pores and funnels. The pores are paired and are in the

posterior part of the 6th somite about midway between a line joining the

ventral seta bundles and one joining the dorsal seta bundles of the 6th

and 7th somites (figs. 1 & 14). No distinct funnels could be identified

comparable with those figured and described by Piguet ('09) for Nais

obiusa, although there appeared to be several cells on each side which

might properly be interpreted as belonging to a funnel since they are

sharply dififerentiated from the clitellum (fig. 15). The cells of the

clitellum are so graded in length as to form a distinct depression with the

funnel cells at the base. The paired funnels are in the same relative

position as those of A^ais obiusa. The lumen of the pore could not be

located, probably because of the thickness of the transverse sections, and,

for the same reason, the opening in the muscular layers was not observed.

The muscular layers were found separated from the pore cells in that

region. The funnel and pore would no doubt be much more conspicuous

at the time of emission of the eggs.

5 6 7 ^'- R

Fig. 1. Diagram showing the general plan of arrangement of the sex organs in somites

5 to 8. Reconstructed from camera lucida outlines of serial transverse sections. Lettering

as for other figures. The numbers indicate the position of the setae bundles. 120X.

Ovisac, (fig. 1). The ovisac is formed by septum 6/7 and extends

posteriad, in one specimen through the 12th somite, in another, just be-

yond the seta bundles of the 9th. Its extent is no doubt dependent upon

the quantity of ova present. It almost fills the body cavity for the major

part of its length, and is distended with a granular appearing material

which stains pink with eosrn, but contains no ova. The sperm sac lies

within its anterior portion.

Ovaries. No ovaries could be found. They should be located on the

posterior side of septum 5/6 since the ovisac extends posteriad from 6

and the oviducal pore is in the posterior part of this somite (fig. 1). Their

absence is no doubt due to the advanced stage of development of the speci-

mens. The absence of ova and the presence of abundant sperm cells

suggests the possibility that an interval of time elapses between the

164 ROY L. MAYHEW

functioning of the spermaries and ovaries. However this does not seem

probable because of the very extensive development of the ovisac. It

seems more probable that ova have occupied the latter and have been

discharged.

Piguet ('06) refers to the appearance of gonads as follows: "Michael-

sen suppose que, chez les Na'ididees, les gonades disparaissent entierement

avant le developpement des autres organes genitaux; cela est sans doute

vrai en general, mais il pourrait y avoir la une exception. Frank Smith

(1896, PI. 35, fig. 4, t.) figure un reste de testicule chez Pristina Leidyi."

In several mature specimens of Paranais uncinata Piguet has observed

vestiges of ovaries. It seems, therefore, that there are individual excep-

tions, but the few observations that have been possible upon Dero limosa

indicate that gonads are developed only during the period of production

of the germ cells.

LITERATURE CITED

BousFiELD, Edward C, 1887. The Natural History of the Genus Dero. Jour. Linn. Soc.

Zool. London, 20:91-106. 3 pi.

MiCHAELSEN, W., 1900. Oligochaeta. Das Tierreich. 10. Lief. Berlin.

1903. Oligochaeten (Hamburgische Elb-Unlersuchung 4). Jahrbuch der Hamburgis-

chen Anstalten, 19:169-209. 1 pi.

1909. Oligochaeta. Siisswasserfauna Deutschlands. Heft 13.

Piguet, Emile, 1906. Observations sur les Naididees et revision systematique de quelques

especes de cette famille. Revue Suisse de Zoologie, 14:185-315. 4 pi.

1909. Nouvelles observations sur les Naididees. Revue Suisse Zoologie, 17:171-216.

Ipl.

Walton, L. B., 1906. Naididae of Cedar Point, Ohio. Am. Nat., 40:683-706. 12 fig.

ABBREVIATIONS

DESCRIPTION OF FIGURES

Fig. 2. Transverse section in the 5th somite showing spermathecae. 120X.

Fig. 3. Portion of left spermatheca, its duct and adjacent body wall in transverse

section. 270X.

Fig. 4. Transverse section of si)crmi(lucal funnel with sperm cells. 270X.

Fig. 5. Transverse section of spermiducal funnel posteriad of section represented in

fig. 4. 270X.

ANATOMY OF DERO LIMOSA LEIDY 165

Fig. 6. Transverse section of sperm duct and anterior end of| atrium at the point of

entrance of the duct. 270X.

Fig. 7. Transverse section of the left ventral portion of a specimen at the point where

the sperm duct bends anteriad. 270X.

Fig. 8. Transverse section of the left ventral portion of the same specimen, as

represented in the preceding figures, at the point where the hypodermal invagination receives

the sperm duct. 270X.

Left ventral setae bundle of the 7th somite. 270X.

Left ventral setae bundle of the 30th somite. 270X.

Left ventral seta of the 2nd somite. 270X.

Left ventral seta of the 3rd somite. 270X.

Right dorsal setae bundle on the posterior half of a specimen. 340X.

Diagram showing the position of the oviducal funnels. The position of the

setae of the 7th somite is shown, as obtained by superimposing the sections containing them

(21 sections posteriad) upon the outline of the sections containing the funnels, by means of a

camera lucida. 120X.

Fig. 15. Diagram showing the differentiation of the oviducal funnel from the clitellum

270X.

Fig. 16. Transverse section of the closed branchial pavilion. 150X.

166

ROY L. MAYHEW

spm

PLATE XVI

STUDIES ON AMERICAN NAID OLIGOCHAETES

1. Preliminary Note on Naids of Douglas Lake, Michigan*

By

H. E. Hayden, Jr.

University of Richmond

During July and August, 1921, I was engaged in a study of the Naididae

of the region around the University of Michigan Biological Station on

Douglas Lake in the upper end of the southern peninsula. This locality

afiForded a wealth of material for such a study, including a number of

sexually mature forms. A full discussion of the systematic aspects of this

work is in course of preparation, to be followed by papers dealing with the

morphology and histology of the various naids, especially of the mature

individuals.

As a preliminary report on this work, I wish to place on record here,

for the benefit of other students of this family of the Oligochaeta, a list

of the species noted, together with brief diagnoses of two new species and

notes on two new varieties. The following established species were

represented by forms which did not differ appreciably from the published

descriptions: »

Aulophorus furcatus (Oken)

Chaetogaster diaphanus (Gruithuisen)

Chaetogaster langi Bretscher

Chaetogaster limnaei K. von Baer

Dero liniosa Leidy

Dero perrieri Bousfield

Nais communis Piguet

Nais pseudoobtusa Piguet

Nais simplex Piguet

Nais variabilis Piguet

Pristina longiseta Ehrenberg

Slavina appendiculata (d'Udekem)

Stylaria fossularis Leidy

Stylaria lacustris (L.)

Vejdovskyella comata (Vejdovsky)

'Contribution from the University of Michigan Biological Station, and contributions from

the Biological Laboratory of the University of Richmond, No. 1.

167

168 H. E. HAYDEN, JR.

With the exception of Stylaria fossularis and Vejdovskyella comata, all

these species were found in consideFable numbers. Of these two species

only one individual of each was found. In each case, however, the species

is unique and its characteristics sufficiently pronounced to prevent any

error in the identification.

The following species was represented by forms which showed only one

marked difference from the type.

Pristina acquiseta Bourne

The individuals of this species agreed closely with the description of

Naidium tentaculatum given by Piguet (1906), which species was later

united by the same writer (1909) with Pristina aeqmseta Bourne (1891).

In the forms described by Piguet and Bourne, however, there are pecu-

liarly enlarged setae in the ventral bundles of segment 4; while in the

forms that came under my observation these setae were, with one excep-

tion, on segment 5. Whether this amounts to a varietal difference or

whether the position of these setae is a matter of no importance, remains

to be seen.

The following is, in my opinion, entitled to rank as a variety:

Chaetogaster diaphanns, var. cy clops, var. no v.

This is in most respects similar to the type form of the species, but

differs from it in the presence of a very definite median pigmented body

intimately associated with the brain and strikingly like an eyespot.

The following species have not hitherto been described:

Dero polycardia sp. nov.

Worms quite large, 7-10 mm. in length, about 300 microns in diameter.

Color reddish. Swimming actively. Ventral setae of segments 2-5, four

to six in number, about 135 microns long, nodulus proximal, distal tooth

longer than proximal and with a slight swelling at base. Ventral setae

of other segments, four to six in number, about 95 microns long, nodulus

a trifle distal, teeth about equal, distal tooth half as thick as proximal,

and with a slight swelling at base. Dorsal setae beginning on segment 6,

with one or two capilliform setae, somewhat longer than the diameter of

the body, and one or two needle-like setae, about 87.5 microns long, slender,

bifid, nodulus distal, di!?tal tooth longer than proximal, proximal part of

the seta almost straight, distal part strongly curved. Contractile trans-

verse vessels ("hearts") up to eight pair, in segments 6-13 inclusive,

though one or more of the last few pair may be lacking. Blood quite

red. Intestinal dilation in segments 9 and 10. First nephridia in segment

7. Respiratory bursa with dorsal lip, consisting of a median portion and

two lateral ciliated processes. Gills, two pair, of the pyramidal type.

Budding takes place between segments 25 and 36. Sexually mature forms

STUDIES ON AMERICAN NAID OLIGOCHAETES 169

not yet observed. Habitat, in felted masses of blue-green algae attached

to slightly submerged logs in a marshy pond near Burt Lake, Michigan.

Haemonais ciliata sp. nov.

Worms large, as much as 16 mm. in the case of double chains, but able

to contract to about one third of their length. Diameter, about 500

microns. Very active and, because of their rapid contractions and ex-

pansions, rather leech-like in their movements. Color, light reddish.

Number of segments up to 55 in individual worms, and up to 100 in double

chains. Prostomium rather acuminate; when expanded, slightly longer

than broad at the base. Eyes absent. Prostomium covered with fine,

straight tactile processes; a zone of similar processes around each segment.

Remainder of body surface bears frequent smaller processes which are

sharply reflexed and terminate in a bulbous swelling. As far back as the

first segment bearing dorsal setae, body surface ciliated. Setae about

middle of segment. Ventral setae usually three in number, about 90

microns long, sigmoid, nodulus about middle, teeth equal in length, distal

tooth half as thick as proximal, and with a slight swelling at base. In all

the individuals observed, nine in number, the first four or five segments

were very short, and the setae of these segments, while having the same

form as those following, v/ere relatively smaller. Dorsal setae beginning

on any segment from 14 to 22 inclusive: with one capilliform seta, about

160 microns long, slightly sigmoid, distal half more curved than proximal;

and one biuncinate seta, about 110 microns long, slightly sigmoid, nodulus

barely distal, teeth long, distal tooth longer and a trifle thinner and with

a very slight swelling at base. Pharynx short, pigmented at both ends.

Remainder of canal not highly specialized. Contractile transverse vessels

("hearts") in most of segments 4-20 inclusive. Circulatory system more

like the usual naid type than that of H. waldvogeli Bretscher (1900).

Budding takes place after segment 40. Mature forms not yet observed.

Habitat, in felted masses of blue-green algae attached to slightly sub-

merged logs, and in water-macerated wood, from a marshy pool near

Burt Lake, Michigan. In Bretscher's (1900) description of H. waldvogeli,

no mention is made of the presence of cilia on the body surface, and as

this is such a noticeable feature of H. ciliata, I have ventured to indicate

this fact in the specific name.

The attention of systematic zoologists is called to the existence in

this country of representatives of two genera not given in the key to the

Naididae on pages 638-640 of Ward and Whipple's "Fresh- Water Biology":

namely, Haemonais and Vejdovskyella. The latter will be found in

Michaelsen (1909), as well as, under the older name Bohemilla, in Michael-

sen (1900). Haemonais, hitherto known only through the single species,

H. waldvogeli, is described in Bretscher (1900). These two genera may be

170 H. E. HAYDEN, JR.

added to the key in Ward and Whipple by altering the text of page 639

as follows:

10 (11) Setae of dorsal bundles all uncinate

Paranais Czerniavsky 18S0.

11 (10) Dorsal setae nearly straight, slightly toothed or simple-

pointed OpJiidonais Gervais 1838.

12 (9) Capilliform setae present in dorsal bundles 13.

13 (13J/2, 21) First anterior dorsal setae on XII to XXII

Haemonais Bretscher 1900.

133^ (13, 21) First anterior dorsal setae on V or VI 14.

14 (18) Posterior end not modified into a gill-bearing respiratory

organ 15.

15 (I53/2) Capilliform setae of dorsal bundle with a series of very

prominent teeth; first anterior dorsal setae on V.

VcjdovskyeUa Michaelsen 1903

1534 (16, 17) Capilliform setae without teeth; one or more capilliform

setae of VI much longer than those of other somites

and equal to three or four times the diameter of the

body Slavina Vejdovsky 1883.

16 (153/^, 17) Prostomium elongated to form a proboscis; dorsal setae

of VI similar in length to those of other somites

Siylaria Lamarck 1816.

17 (153/2. 16) Without proboscis; dorsal setae of VI similar in length to

those of other somites Nais Miiller 1774.

18 (14) Posterior end modified into a gill-bearing respiratory organ,

the branchial area 19.

19 (20) Ventral margin of the branchial area with a pair of long

processes Aulophorus Schmarda 1861.

Two ecological notes may be made very briefly here. The observation

of Mrazek (1917) as to the ingestion of trematode larvae by Chaciogaster

limnaei is similar in all respects to observations made in the course of my

study of this form in Michigan. Chactogaslcr is in general carnivorous,

especially Ch. diaphanns. This latter species, particularly the cyclops

variety, is actively predaceous and even cannibalistic, and those who are

just beginning the study of these worms are warned to keep their chaeto-

gasters away from vessels containing other genera, as they will depopulate

a culture of naids in a very little time.

BIBLIOGRAPHY

Bourne, A. G.

1891. Notes on the Naidiform Oligochacta, etc., Quart. Jour. Micro. Sci. (n. s.),

32:335-356.

STUDIES ON AMERICAN NAID OLIGOCHAETES 171

Bretscher, K.

1900. Mittheilungen iiber die Oligochaetenfauna der Schweiz. Rev. Suisse de Zool.,

8:1-44.

MiCHAELSEN, W.

1900. Oligochaeta, Das Tierreich, 10. Berlin.

1909. Oligochaeta, Die Sussvvasserfauna Deutschlands, 13. Jena.

Mrazek, a.

1917. The feeding habits of Chaetogaster limnaei. Sbornik Zoologicky, 1 :22-23. Prague.

PiGUET, E.

1906. Revision des Naididae. Rev. Suisse de Zool., 14:185-315.

1909. Nouvelles observations sur les Naididees. Rev. Suisse de Zool., 17:171-218.

Ward, H. B. and Whipple, G. C.

1918. Fresh-Water Biology. New York.

EXCESSIVE SEXUAL DEVELOPMENT IN HYDRA OLIGACTIS

WITH SPERMARY ON TENTACLE

By

Arthur W. Schmidt

Department of Zoology, University oj Nebraska^

This specimen of Hydra oligactis was found in an aquarium with other

normal individuals of the same species in February of 1918. The culture

was abundantly supplied with food, such as Daphnia, Cyclops, and various

other Crustaceans. The specimen was killed with a solution of corrosive

sublimate and acetic acid, stained in a special preparation of borax car-

mine, and mounted in Canada balsam. It measures 11 mm. in length,

the body 83^ mm. and the tentacles 2}/^ mm. as it is miounted on the

slide. Its extreme length when living was 13 mm. Fifteen spermaries

and two ovaries occur on the body and one spermary on one of the

tentacles of the specimen (See figure). The spermary on the tentacle

appears to be perfectly normal in all respects except location. The

question now arises as to the cause of its occurrence on the tentacle.

Parke cites the following instances in the establishment of normal

tentacles from abnormal ones. "A nine-tentacled Hydra fusca with one

branching tentacle was isolated on February 27th. On February 28th one

branch had revolved about 45° till it was in line with the longitudinal axis

of the tentacle, while the other one appeared somewhat shorter than it

did at first. On March 1st the small branch was almost entirely resorbed.

It was much nearer the end of the tentacle than before and appeared as a

small outgrowth from the tentacle. This apparent shifting of the small

branch, by a migration of the short branch, may have taken place in three

ways: by a shortening of the long branch, by a migration of the short

branch towards the end of the tentacle, or by a fusion of the two branches

along the median line. The first or last explanation seems_ the most

plausible since similar instances were seen in which there could be no doubt

that these were the processes involved. On March 3rd the small branch

was entirely resorbed, leaving the Hydra with nine normal tentacles. Two

other instances of regulation of forked tentacles in the same manner were

observed." He states further that his observations show that branching

tentacles may arise by the fusion of two tentacles and may regulate them-

selves by a complete fusion along their median sides so as to form a single

tentacle.

* Studies from the Zoological Laboratory, The University of Nebraska, No. 130.

172

SEXUAL DEVELOPMENT IN HYDRA OLIGACTIS

173

In further observations he found that by regulative processes two

distinct tentacles could be reformed out of two fused tentacles. He

cites several instances in which two of the tentacles were fused near their

ends and not near their bases along the median line. "This fusion,"

he states, "may have been caused by an injury to one of the tentacles,

the other tentacle having become attached to it. This seems probable

from the fact that alternate tentacles as well as adjacent tentacles were

found fused in this manner, One eight-tentacled PI\dra was found

Hydra oligac is: A, showing 18 gonads; s, spermary; o, ovary; d, basal disc. B, showing

position of spermary on tentacle.

in which two alternate tentacles had stuck together at a point about three

fourths of the distance from the base to the tips of the two tentacles at

the point of fusion. There was no connection between the cavities of the

two tentacles at the point of fusion. The next day after the Hydra had

been isolated, one of the tentacles had constricted off from the other

tentacle just below the point of fusion of the two, leaving the tip of its

tentacle attached to the other branch. The cavities of the two branches

174 ARTHUR W.'^SCHMIDT

were in direct communication. The process of regulation that now took

place was exactly as in the Hydra described above. This is a good ex-

ample of how branching tentacles may originate." Both tentacles became

normal. He summarizes his conclusions with the statement: "It appears

that three regulative processes may take place in the establishment of

normal tentacles; viz. (1) fusion, (2) resorption, (3) constriction."

The theories of fusion and constriction suggest the idea that by a

contact of the tentacle with the spermary in its original position on the

body of the Hydra the two were fused; and then by a process of constric-

tion the spermary was severed from the body, leaving it in its present

position on the tentacle. Parke's reference to the migration of a short

branch of a tentacle, mentioned above, also suggests the idea that the

spermary migrated from an original position on the body to its present

position on the tentacle. Probably the most plausible suggestion, however,

is that the position of the spermary is due to an unusual local stimulation

of a group of interstitial cells in the ectoderm of the tentacle, causing ab-

normal development in that location. Due, however, to the absence of

facts regarding previous conditions in the aquarium, it is impossible to

throw any light upon the real cause of this abnormal condition, except by

way of suppositions from previous investigations.

Whitney, in his observations on Hydra viridissima, found that when

they are subjected to a low temperature and starvation they develop testes

and eggs. Hertwig, however, found that if Hydra oligactis is kept at a

temperature^ of 8°-10° C. it will develop testes irrespective of the food con-

ditions. At the time of the discovery of this specimen of Hydra oligactis

that was about the temperature of the room in which the aquarium that

contained this specimen was kept.

Extensive experimental work on sexually reproducing Hydras would

undoubtedly reveal the generality or abnormality of this occurrence.

The writer desires to express his thanks to Drs. Robt. H. Wolcott

and David D. Whitney for access to materials and laboratory facilities,

also for their suggestions and generous interest.

BIBLIOGRAPHY

Hertwig, Richard. 1906 tjber Knospung und Geschlechtsentwickelung von H\c]ra fusca.

Biol. Centralbl., Bd. XXVI, No. 16, pp. 489-507.

Parke, H. H. 1900 Variation and Regulation of Abnormalities in Hydra. .Arch. f. Entw.-

Mech., Bd. X, Heft 4, pp. 692-710.

Peebles, Florence. 1900 Experiments in Regeneration and Grafting of Hydrozoa. Arch. f.

Entw.-Mech., Bd. X, Heft 2 & ?>, pp. 435-487.

Rand, Herbert W. 1899 Regeneration and Regulation in Hydra viridis. Arch. f. Entw.-

Mech., Bd. VHl, Heft 1, pp. 1-34.

Whitney, David Day. 1907 The Influence of External Factors in Causing the Development

of Sexual Organs in Hydra viridis. Arch. f. Entw.-Mech., Bd XXIV, Heft 3,

pp. 524-537.

SOME SUGGESTIONS FOR TEACHING MYCOLOGY

By

F. D. Heald

The study of any group of plants as to its taxonomy may proceed

along two widely divergent lines: First, the student may be taught to

use artificial keys and determine species, which are put in their respective

species pigeon-holes and properly labeled, the prime object being to deter-

mine the binomial, but little concern being given to the relationship of the

various forms studied; or Second, the student may be taught to construct

diagrammatic keys to the various groups, which will express natural rela-

tionship. These natural keys, which give a graphic representation of

relationship, are clearer than the obscurely worded artificial keys crowded

full of technical terms. The writer has followed the latter method with

marked success in presenting the taxonomy of seed plants with successive

classes through a period of years and more recently has used the same

plan with classes in mycology. The method has a number of features to

recommend it, some of which are: (1) The creation of a greater interest

on the part of the student in his work; (2) The development of the stu-

dents' ability to reason and weigh evidences; (3) The cultivation of the

scientific imagination; (4) A better understanding of evolution and what

it means; and (5) The possibility of emphasizing natural descent of the

various groups and bringing out the fact that classification is in reality

but a means to an end,^ — an expression of relationships.

The work in mycology in our laboratory is offered to students who

have had general elementary botany and also to those who have had in

addition a semester in general pathology, in both of which they gain some

familiarity with fungi. The minimum time which suffices for anything

like a satisfactory presentation of the subject is six hours of laboratory

work throughout the year. The general method of procedure may be

briefly presented.

Very early in the beginning of the work the class is given a skeleton

outline of the great groups, somewhat as shown in the accompanying

diagram (Fig. 1), except that the diagrams are omitted and the student is

required to select diagrams and make any adjustments that may seem

necessary to present concepts of the great groups by the visual channel,

or to bring out more clearly the natural relationships. Various mycological

works, such as Engler & Prantl, Rabenhorst's Kryptogamen Flora,

special monographs, etc. must be available for reference. Suggestions are

175

176

F. D. HEALD

given to the student, but they are encouraged to use their own originality

and independent thought as well as to consult authorities. They are also

,.-~^

16.HYMENJ0MYCETALES *"

^^ /'' 14.EX0BAS1D1ALES '

15.GASTER0MYCETALES

^*'- ALLIES

13. AURICU L ARIALES ^^''ALLl E S

12.USTILAGINALES

MYCELIUM SEPTATE

I I

ll.UREOINALES

10. TUBERALES •'^ALLIES

9. pezizalesAllies

7.sphae:riales»*''allies

6. PERISPORIALES /ALLIES

MONILIkLCS

8.FUNGI IMPERFECTI

5. EXOASCALES ^'^'ALLIES

MYCELIUM NON-SEPTATE

yio %

lOWNV MILOCWS'

|(? tiOo#e.

I.SCHIZOMYCETES

BlCrcHiA

/5>V

.4. OOMYCETES.

TT 7 ^vnnMvr-irTrc

3. ZYGOMYCETES

-^.PRIMITIVE FORMS --'

i,, MYXOMYCETES

si-'xi Ht^n

Fig. 1. Chart ShowinR the (Jrcat Group of Fungi.

SOME SUGGESTIONS FOR TEACHING MYCOLOGY 177

given the understanding that schemes of natural relationship can be

nothing more than the expression of individual opinion, which should be

arrived at by weighing all the evidence that can be brought to bear in any

specific case. No two students will choose the same illustrations and the

more intimate details of relationship as expressed in the charts are certain

to show variations. These variations afTord excellent material for class

discussions which can frequently be held with much profit.

The logical order for the study of the great groups would be to begin

at the bottom of the family tree with the most primitive forms and proceed

to the more complex and higher forms later. In actual practice, however,

it seems better to sacrifice logic and begin with some group which more

readily lends itself to the method in question. The Erysiphaceae, or

powdery mildews of the order Perisporiales, is a family well suited to

introduce the plan of study: (1) Because species determination is relatively

easy; (2) Because representatives of all the genera can very readily be

obtained in most environments. Our plan would call for a careful and

detailed study of some type of each genus, accompanied by drawings.

Following this the student is asked to construct a diagrammatic key to the

genera of powdery mildews, which will express relationship and afford

generic concepts, mainly through the visual channel. Before this key is

made, a general class discussion is held and the more important characters

which may indicate relationship are briefly reviewed, with emphasis on

those which are primitive and those which are more advanced. These

keys are then presented for comparison and discussion (Fig. 2). After the

completion of the keys, the class is asked to determine the species of all

the powdery mildews which they have collected on some of their special

field trips.

T^OtXBPMACRA

Fig. 2. Chart Showing the Genera of Powdery Mildews

Essentially the same plan is followed with all the great groups or

alliances, or with representative families of these groups. It will be at

once evident that all groups can not be treated as fully as the Erysipha-

178 F. D. HEALD

ceae, which we have used for our introduction to the method. For

example, in the study of the Sphaeropsidales of the Imperfect Fungi,

attention is given to the genera furnishing parasites and only these are

included in the graphic key which the students are required to construct.

In other cases, as in the Sphaeriales and allies with numerous families, the

graphic keys may be limited to a representation of the families.

As previously stated, the minimum time for a course in mycology

according to the plan outlined is six hours of laboratory work per v/eek

throughout one school year. With this minimum time there must of

necessity be many omissions and consequently much of the success of the

course depends on the judgment of the instructor in making wise selec-

tions. There is no doubt that the same plan could be followed with much

profit throughout an additional year of work.

This brief note has been prepared at the suggestion of several of my

former students who have been stimulated to further mycological study

by the use of the method outlined. It is hoped that it may offer some

suggestions to some of our younger mycologists who have received their

instruction by the pigeon-hole method.

Department of Plant Pathology,

Washington Stale College,

Pullman, Washington.

LIST OF MEMBERS

Honorary Members

Crisp, Frank, LL.B., B.A., F.R.M.S 5 Landsdowne Road, Netting Hill, London, Eng.

Pflaum, Magnus 2334 S. 21st St., Philadelphia, Pa.

Life Members

Brown, J. Stanford, Ph.B., A.M P.O. Box 38, Far View, Black Hall, Conn.

Capp, Seth Bunker P.O. Box 2054, Philadelphia, Pa.

Duncanson, Prof. Henry B., A.M R.F.D. 3, Box 212, Seattle, Wash.

Elliott, Prof. Arthur H 52 E. 41st. St., New York City.

Hately, John C Chicago Beach Hotel, Chicago, 111.

Members

The figures denote the year of the member's election, except '78 which marks an original

member. The TRANSACTIONS are not sent to members in arrears, and two years'

arrearage forfeits membership. (See Article IV of By-Laws.)

Members .\dmitted Since the Last Published List

BucHHOLz, J. T. Linton, Edwin

Challis, Frank E. McCauley, David V.

Cheavin, William S. Noland, Lowell E.

Faust, E. C. Plunkett, Orda A.

Hartman, Ernest Ryan, Ruth

Harris, D. F. Titus, C. P.

Jewell, Mina E. Young, Paul A.

Kamm, Minnie Watson Zimmerman, Naomi B.

List of Members

Ackert, James Edward, Ph.D., '11 Kas. State Agr. Col., Manhattan, Kans.

Adams, Frederick, C. E., '19 Apartado 560, Mexico, D.F., Mexico.

Allen, Harrison Sanborn, M.A., '15 442 Farmington Ave., Waterbury, Conn.

Allen, Wm. Ray, M.A., '15 Dept. Zoology, Univ. of Kentucky, Lexington, Ky.

Allen, Wynfred E., A.M., '04 Scripps Inst., La Jolla, Calif.

Anderson, Emma N., '16 Station A, Lincoln, Nebraska.

Andras, J. C, B.A., '12 540 S. Main St., Manchester, 111.

Arnold, L. P., O.D., '20 Vulcans Temple, Carlisle, Ark.

Arnold, Wm. T., '17 21 Park Rd., Wyomissing, Pa.

Ashley, Frank M., M.E., '20 Tribune Building, New York City, N. Y.

Atchison, Mrs. W. S., A.M., '16 263 Walnut Ave., Elgin, 111.

Atherton, Prof. L. G., A.B., M.S., '12 State Normal School, Madison, S. D.

Atwood, H. F., '78 16 Seneca Parkway, Rochester, N. Y.

Baldwin, Herbert B., '13 927 Broad St., Newark, N. J.

Barker, Fr.anklin D., PH.D. '03 Univ. of Nebraska, Lincoln, Nebr.

Barre, H. W., B.Sc., M.A., ^2 Clemson College, S. C.

Bausch, Edward, '78 179 N. St. Paul St., Rochester, N. Y.

179

180 LIST OF MEMBERS

Bausch, William, '88 St. Paul St., Rochester, N. Y.

Bean, A.M., M.A., '15 1501 Palm Ave., Fresno, Calif.

Beck, William A., M.Sc, '16 Univ. of Dayton, Dajton, Ohio.

BiCKNELL, Anna (Miss) B.S., '21 1713 Lamont St., X.W., Washington, D. C.

BiERBAUM, C. H., '21 Mutual Life Bldg., BufTalo, N. Y.

BiRGE, Prof. E. A., Sc.D., LL.D., '99 772 Langdon St., Madison, Wis.

Black, J. H. M.D., '12 530 Wilson Bldg., Dallas, Texas.

Bo\T)EN, Alan Arthi^r, '21 1421 Oakridge Ave., Madison, Wis.

Boyer, C. S., A.m., '92 6140 Columbia Ave., Philadelphia, Pa.

Brown, Alice L., '19 Kans. St. Ag. Col., Manhattan, Kans.

Brunn, Ch.arles a., LL.B., '16 314 Reliance Bldg., Kansas City, Mo.

Bryant, Prof. Earl R., A.M., '10 Muskingum College, New Concord, O.

BucHHOLz, Prof. John T., Ph.D., '22 Dept. of Botany, Univ. of Arkansas, Fayetteville, Ark.

Bltfalo Society of Natural Sciences Library Building, BufTalo, N. Y.

Bull, James Edgar, Esq., '92 141 Broadway, New York City.

Bullitt, Prof. J. B., M.A., M.D., '12 Chapel Hill, N. C.

BuswELL, A. M., M.A., '16 Univ. of Illinois, Urbana, III.

Caballero, Prof. Gust.w A., '16 Fordham Univ., New York City.

Carlson, C. O., A.B., '13 Doane College, Crete, Nebr.

C.A.RTER, Prof. Charles, 'U Parsons College, Fairfield, la.

Carter, John E., '86 5356 Knox St., Germantown, Philadelphia, Pa.

Challis, Frank E., '22 252 Central Ave., Aurora, 111.

CHEA^^N, William Squier, F.R.M.S., F.E.S. (Lond.), F.C.S. (Lond.), '22

^liddlesex Hosp. ^led. School, London, W. England.

Chester, Waylant) Morgan, M.A., '15 Colgate University, Hamilton, N. Y.

Chickering, A. M., A.M., '16 Albion, Michigan.

Clark, George Edw., M.D., '96. . Sheppard & Enoch Pratt Hospital, Towson, Maryland.

Cl.^rk, Howard W., A.M., '12 Fairport, Iowa.

Cleveland, L. R., B.S., '21 310 W. Monument St., Baltimore, Md.

Cobb, N. A., Ph.D., '14 Falls Church, Va.

Coghill, Prof. George E., Ph.D., '11 R.F.D. 9, Lawrence, Kans.

Colton, Harold S., Ph.D., '11 Zoological Lab., Univ. of Pa., Philadelphia, Pa.

Cone, Albert, '12 . .Associate Editor, Lumber World Review, 608 So. D arborn St., Chicago,

lU.

Conger, Allen C, M.A., '15 527 Forest St., East Lansing, Mich.

Conlon, James J., Ph.D., '14 717 Hyde St., San Francisco, Cal.

Cornell Univ. Library (Prof. S. H. Gage) Ithaca, N. Y.

CoRT, W. W., Ph.D., '11 . . J. H. U. School of Hygiene, 310 W. Monument St., Baltimore, Md.

Covey, George W., '11 2017 South 26th St., Lincoln, Nebr.

Danheim, Miss Bertha L., B.S., '21 Blue Rapids, Kansas.

Darbaker, Leasxjre Kline, Ph.D., M.D., '11

7025 Hamilton Ave., Homewood Sta., Pittsburgh, Pa.

Davis, Prof. H. S., Ph.D., '12 Bureau of Fisheries, Washington, D. C.

Dayton, Miss Edna B., M:D., '21 1512 N. Gratz St., Philadelphia, Pa.

Deere, Emil Olaf, A.M., S.M., '13 Bethany College, Lindsborg, Kans.

Depew, Ganson, '21 167 Summer St., BufTalo, N. Y.

De puy, Percy Leroy, B. S., '19 Federal Bldg., El Reno, Okla.

Diago, Dr. Joaquin, '21 Aguila 72, Havana, Cuba.

Disbrow, William S., M.D., Ph.G., '01 151 Orchard St., Newark, N. J.

Dodge, Carroll W., Ph.D., '14. .Dept. Cr>'ptogamic Botany, Harvard Univ., Cambridge,

Mass.

LIST OF MEMBERS 181

DoLBEY, Edward P., '06 3613 Woodland Ave., Philadelphia, Pa.

DouBLEDAY, ARTHUR W., M.D., '16 5 Marlborough St., Boston, Mass.

Drescher, W. E., '87 Care Bausch & Lomb Opt. Co., Rochester, N. Y.

Duncan, Prof. F. N., Ph.D., '16 So. Methodist Univ., Dallas, Tex.

Edmon'dson, Charles H., Ph.D., '15 College of Hawaii, Honolulu.

Eggleston, H. R., M.A., '13 Marietta College, Marietta, Ohio.

Eigenmann, Prof. C. H., '95 630 Atwater Ave., Bloomington, Ind.

Elliott, Frank R., M.A., '15 324 Kinsey St., Richmond, Ind.

Ellis, Prof. M. M., Ph.D., '12 Dept. Physiol., University of Mo., Columbia, Mo.

Elmore, Prof. C. J., '19 706 West St., Emporia, Kan.

Elrod, Prof. Morton J., M.A., M.S., '98 University of Montana, Missoula, Mont.

Enburg, J. M., '20 5207 Baltimore St., Philadelphia, Pa.

Essenberg, Mrs. Christine, M.S., '16 Scripps Institute, La Jolla, Cal.

Esterly, Calvin O., '15 Occidental College, Los Angeles, Cal.

Eyre, John W. H., M.D., M.S., F.R.M.S., '99. .Guy's Hospital, London, S. E., England.

Fattig, Prof. P. W., B.S., M.S., '12 207 Pine St., Farmville, Va.

Faust, Ernest C, Ph.D., '22 Peking Union Medical College, Peking, China.

Fellows, Chas. S., F.R.M.S., '83 107 Cham, of Comm., Minneapolis, Minn.

Fellows, Harriette L., '21 220 S. Prairie Ave., Siou.x Falls, South Dakota.

Fernandez, Fr. M.anuel, B.S., '16 San Juan de Latran College, Manilla, P. I.

Findlay, Merlin C, A.M., '15 Park College, Parkville, Mo.

FooTE, J. S., M.D., '01 Creighton Dental College, Omaha, Nebraska.

Furniss, H. W., M.D., Ph.D., '05 56 Brazos St., West Hartford, Conn.

Gabriele, H. J., '16 2659 California St., San Francisco, Cal.

Gage, Prof. Simon H., B.S., '82 Stimson Hall, Ithaca, N. Y.

Galloway, Prof. T. W., A.M., Ph.D., '01

Penn Terminal Bldg., 370 Seventh Ave., New York, N. Y.

Gilbert, E. M., Ph.D., '19 Biology Building, U. of Wis., Maidson, Wis.

Goldsmith, G. W., B.A., '13 123 E. Washington, Colorado Springs, Colorado.

Gowen, Francis H., '14 R. D. 1, Box 14, Exeter, N.H.

Graff, John H., '19 Research Dept., Brown Company, Berlin, N. H.

Graham, Charles W., M.E., '11 1033 Mills Bldg., San Francisco, Cal.

Graham, John Young, Ph.D., '14 University, Alabama.

Gravelle, p. O., '19 114 Prospect St., South Orange, N. J.

Griffin, Lawrence, E., '13 Reed College, Portland, Ore.

Gross, F. O., M.D., '19 1816 Erie Ave., Philadelphia, Pa.

GuBERLET, John E., Ph.D., '11 A. & M. College, Stillwater, Okla.

GuNNS, Cecil Aguila, '21. .Dept. Zoology, Kansas State Agr. College, Manhattan, Kansas.

GuYER, Michael F., Ph.D., '11 University of Wisconsin, Madison, Wisconsin.

Hagelstein, Robert, '16 165 Cleveland Ave., Mineola, Nassau Co., N. Y.

Hague, Florence, A.M., Ph.D., '16 . .Dept. Zoology, Oregon Agr. Coll., Corvallis, Oregon.

Hall, F. Gregory, B.A., '17 Biology Building, Univ. of Wis., Madison, Wis.

Hallinen, J. E., B.S., '21 Cooperton, Okla.

Hance, Robert T., B.A., '13 Zool. Lab., N. Dakota Agr., College, North Dakota.

Hankinson, T. L., B.S., '03 St. Normal School, Ypsilanti, Michigan.

Hansen, James, '15 St. Johns Univ., Collegeville, Minn.

Hardy, Eugene H 1230 S. Keystone Ave., Indianapolis, Ind.

Harman, Mary T., Ph.D., '13 Kansas State Agr. College, Manhattan, Kansas.

Hartman, Ernest, B.S., '22 Kans. St. Agr. College, Manhattan, Kansas.

Harris, David Eraser, M.D., D.Sc, F.R.S.S., '22 Dalhousie University, Halifax, N. S.

Hayden, H. E., A.m., '21 Department of Biology, Univ. of Richmond, Va.

182 LIST OF MEMBERS

Hayes, W. P., M.S., '19 319 N. 18th St., Manhattan, Kans.

Heald, F. D., Ph.D., '06 Wash. State College, Pullman, Wash.

Heath, Roy Franklin, M.Sc, '18 P.O. Box 270, Billings, Montana.

Henderson, William, '11 Mellon Inst., Univ. of Pittsburgh, Pittsburgh, Pa.

Herrick, Chester A., B.S., '21 Kansas State Agri. College, Manhattan, Kansas.

Hickman, J. R., A.B., '19 Bristol, West Virginia.

Hilton, William A., Ph.D., '15 Claremont, Cal.

Hisaw, F. L., M.S., '19 Kans. State Agr. College, Manhattan, Kans.

Holy Cross College, Professor of Biology Worcester, Mass.

Hopkinson, D., M.D., '20 1008 Third St., Milwaukee, Wis.

Hoskins, Wm., '79 49 6th St., LaGiange, 111.

Hottes, C. F., Ph.D., '20 Nat. Hist. Bldg., Univ. of 111., Urbana, 111.

Hubert, H. E., B.S., '20 3615 Melpomene St., New Orleans, La.

Hltdson, D. v., B.S., '20 Johns Hopkins Medical School, Baltimore, Md.

Ives, Frederic E., '02 1327 Spruce St., Philadelphia, Pa.

Jackson, F. S., M.D., '19 Mc Gill University, Montreal, Canada.

Jacot, A. P., A.B., '19 Biology Dept., Shantung Univ., Tsinanfie, Shantung, China.

Jeffs, Prof. R. E., '11 624 N. Johnson St., Iowa City, Iowa.

Jewell, Mina E., Ph.D., '22.. Dept. Zoolog>', Kans. St. Agr. College, Manhattan, Kansas.

Jordan, Prof. H. E., '12 34 University Place, Charlottesville, Va.

Juday, Chancey, '00 Biology Bldg., U. of Wis., Madison, Wis.

JuDD, H. D,, Opt.D., '19 460 W. Philadelphia Ave., Detroit, Michigan.

Kamal, Mohammed, B.S., '21.. . .Box 223, Kansas State Agr., College, Manhattan, Kansas.

Kamm, Minnie Watson, Ph.D., '22. . .263 Windermere Rd., Walkerville, Ontario, Canada.

Kincaid, Trevor, A.M., '12 University of Washington, Seattle, Wash.

KiRSCH, Prof. Alexander M., M.G., '16 Notre Dame (Univ.), Ind.

Knight, F. P. H., '11 1015 Blondeau St., Keokuk, Iowa.

Kofoid, Charles A., Ph.D., '99. . . .University of California, 2616 Etna St., Berkeley, Cal.

Kostir, W. M., M.A., Ph.D., '20 Dept. Zoology, Ohio State Univ., Columbus, Ohio.

KoTZ, A. L., M.D., '91 302 High St., Easton, Pa.

Krecker, Frederic H., Ph.D., '15 Ohio State University, Columbus, Ohio.

Kudo, R., Ph.D., '20 Dept. Zoology, Univ. of 111., Urbana, 111.

Lambert, C. A., '12 Bank of New South Wales, Warwick, Queensland, Australia.

LaRue, George R., Ph.D., '11 University of Michigan, Ann Arbor, Michigan.

Latham, Miss V. A., M.D., D.D.S., F.R.M.S., '88

1644 Morse Ave., Rogers Park, Chicago, 111.

Latimer, Homer B., Ph.D., 'U 1226 So. 26th St., Lincoln. Nebr.

Lewis, Ivey Foreman, Ph.D., '18 University, Va.

Lewis, Mrs. Katherine B., '89 656 Seventh St., Buffalo, N. Y.

Linton, Edwin, A.B., A.M. (Wash. Jeff) Ph.D. (Yale) '22

1 104 Milledge Road, Augusta, Ga.

Litterer, Wm., A.M., M.D., '06 Nashville, Tenn.

Lofton, Robert Elwood, A.B., '21 Bureau of Standards, Washington, D.C.

Lome, Adolph, '92 289 Westminster Road, Rochester, N. Y.

Longfiliow, Robert Caples, M.S., M.D., '11 1611 22nd St., Toledo, Ohio.

LowDEN, Hugh B., '16 1312 York St., Denver, Colo.

Lowrey, Eleanor C, '19 1826 D. St., Lincoln, Nebr.

Lyon, Howard N , M.D., '84 828 N. Wheaton Ave., Whcaton, 111.

MacGillivray, Alexander D., '12 603 W. Michigan .\ venue, Urbana, III.

MacKay, Alexander H., B.A., B.Sc, L.L.D., F.R.S. Canada, '21

61 Queen Street, Dartmouth, Nova Scotia, Canada

LIST OF MEMBERS 183

Magath, T. B., M.S., Ph.D., M.D., '13 Mayo Clinic, Rochester, Minn.

Manchee, E. D., '19 200 Glen Cairn Ave., Toronto, Can.

Mannhardt, L. a., Ph.B., '21 ... .N. Y. University, Washington Square, New York, N. Y.

Mark, George Henry, M.E., '11 94 Silver St., Waterville, Maine.

Marshall, Collins, M.D., '96 2507 Penn Ave., Washington, D. C.

Marshall, Ruth, Ph.D., '07 Rockford College, Rockford, 111.

Marshall, W. S., Ph.D., '12 139 E. Gilman St., Madison, Wis.

Martland, Harrison S., A.B., M.D., '14 1138 Broad St., Newark, N. J.

Mather, E., M.D., Ph.D., '02 228 Gratiot Ave., Mt. Clemens, Mich.

May, Henry Gustav, Ph.D., '15

Agr. Exp. Sta., Rhode Island State College, Kingston, R. I.

Maywald, Frederick J., '02 222 Grand Ave., Nutley, N. J.

McCauley, David V., S.J., M.A., '22 Fordham University, New York, N. Y.

McCoLLOCH, J. W., B.S.j '19 Kans. Agr. Exp. Sta., Manhattan, Kans.

McGreery, Geo. L., '13 110 Nevada St., Carson City, Nev.

McCuLLOCH, Irene, Ph.D., '20

Dept. Biology, Sophie Newcomb Memorial College, Tulane Univ., New Orleans, La.

McEwAN, A., '15 Fifth Ave., Guarantee Building, 522 Fifth Ave., New York, N. Y.

McKay, Joseph, '84 259 Eighth St., Troy, N. Y.

McKeever, Fred L., F.R.M.S., '06 P.O. Box 210, Penticton, B. C.

McLaughlin, Alvah R., M.A., '15 108 S. 6th St., Columbia, Mo.

McWilliams, John, '14 Lock Box 91, Greenwich, Conn.

Mercer, A. Clifford, M.D., F.R.M.S., '82 324 Montgomery St., Syracuse, N. Y.

Mercer, W. F., Ph.D., '99 200 E. State St., Athens, Ohio.

Metcalf, Prof. Zeno P., B.A., '12 St. College Station, Raleigh, N. C.

Miller, Charles H., '11 Med. School, Johns Hopkins U., Baltimore, Md.

Miller, John A., Ph.D., F.R.M.S., '89 44 Lewis Block, Buffalo, N. Y.

MocKETT, J. H., Sr., '01 2302 Sumner St., Lincoln, Nebr.

Moeller, H., M.D., '07 341 West 57th St., New York, N. Y.

Moody, Robert P., M.D., '07 Hearst Anat. Lab., U. of Cal., Berkeley, Cal.

Morgan, Anna Haven, Ph.D., '16 Mt. Holyoke Coll., So. Hadley, Mass.

MuTTKOWSKi, R. a., Ph.D., '19 Univ. of Idaho, Moscow, Idaho.

Myers, Frank J., '13 15 S. Cornwall Place, Ventnor City, N. J.

NoLAND, Lowell E., M.A., '23 Biology Building, Univ. of Wis., Madison, Wis.

NoRRis, Prof. Harry Waldo, '11 Grinnell, Iowa.

Norton, Charles E., M.D., '11 118 Lisbon St., Lewiston, Me.

OsBORN, Prof. Herbert, M.S., '05 Ohio State University, Columbus, Ohio.

Ott, Harvey N., A.M., '03 Spencer Lens Co., Buffalo, N. Y.

Patrick, Frank, Ph.D., '91 822 West 58th Street, Kansas City, Mo.

Payne, Miss Nellie M., B.S., '21 Division of Entomology, University Farm, St. Paul,

Minn.

Pease, Fred N., '8/ P.O. Box 503, Altoona, Pa.

Pennock, Edward, '79 3609 Woodland Ave., Philadelphia, Pa.

Peterson, Niels Fr3:derick, '11 Plainview, Nebr.

Pickett, F. L., Ph.D., '20 Dept. Botany, State College of Washington, Pullman, Wash.

Piatt, H. S., Ph.D., '19 561 W. 141st St., New York, N. Y.

Pitt, Edward, '11 Brandhock, Gerrard's Cross, Bucks, England.

Plough, Harold H., A.M., Ph.D., '16 Dept. Biology, Amherst Coll., Amherst, Mass.

Plunkett, Orda a., am., '22 Dept. Botany, Univ. of Illinois, Urbana, 111.

POHL, John C, Jr., '17 204 N. 10th St., Easton, Pa.

Pool, Raymond J., Ph.D:, '15 Station A, Lincoln, Nebr.

184 LIST OF MEMBERS

Pound, Roscoe, A.M., Ph.D., '98 Harvard Law School, Cambridge, Mass.

Powers, E. B., A.B., Ph.D., '12

Univ. of Tenn., College of Medicine, 718 Union Ave., Memphis, Tenn.

Praeger, Wm. E., M.S., '14 421 Douglas Ave., Kalamazoo, Mich.

Procter, William, Ph.D., '19 Suite 423, 30 East 42nd St., New York, N. Y.

PuRDY, William C, M.Sc, '16 1311 Burdette Ave., Cincinnati, Ohio.

PuTZ, Alfred, '21 5117 Locust St., Philadelphia, Pa.

Qulllian, Marvin C, A.M., '13 .• Wesleyan Col., Macon, Ga.

Rankin, Walter M., '13 Princeton University, Princeton, N. J.

Ransom, Brayton H., '99 U. S. Bureau of Animal Industry, Washington, D. C.

Reese, Prof. Albert M., Ph.D., (Hop.) '05 W. Va. Univ. ,Morgantown, W. Va.

Richards, Aute, Ph.D., '12 Dept. Zoology, Univ. of Oklahoma, Norman, Oklahoma.

Riley, C. F. Curtis, M.S., '15 Univ. of Manitoba, Winnipeg, Can.

Roberts, E. Willis, '11 65 Rose St., Battle Creek, Mich.

Roberts, J. M., '11 460 E. Ohio St., Chicago, 111.

Robinson, J. E., M.D., '15 Box 405, Temple, Texas.

Roe, G. C, A.B., '17 113 R. Street, N.W., Washington, D. C.

Rogers, Walter E., '11 Lawrence College, Appleton, Wis.

Root, Francis Metcalf, Ph.D., '21 310 W, Monument St., Baltimore, Md.

Ross, Luther Sherman, S.M., '11 1308 27th St., Des Moines, Iowa.

Rush, R. C, M.D., '12 Hudson, Ohio.

Ryan, Ruth, A.B., '22 Dept. Botany, Univ. of Illinois, Urbana, 111.

Rye, L. E., A.B., '20 36 Lincoln Rd., Brockton, Mass.

ScHEAR, E. W., E., '19 107 West Park, W^estervUle, Ohio.

Scott, Helen M., A.M., '19 1002 West St., Grinnell, Iowa.

Scott, J. W., Ph.D., '12 Univ. of Wyo., Laramie, Wyo.

Shearer, J. B., '88 809 Adams St., Bay City, Mich.

Sheerar, Leonard F., '19 158 W. State St., Wellsville, N. Y.

Sheldon, John Lewis, Ph.D., '15 Morgantown, W. Va.

Simon, C. L., M.D., '20 1734 Linden Ave., Baltimore, Md.

Smith, Prof. Frank, A.M., '12 1005 W. California Ave., Urbana, 111.

Smith, Gilbert Morgan, Ph.D., '15 113 Spooner St., Madison, Wis.

Soar, C. D., F.R.M.S., '07 10 West Moreland Rd., Barnes, London, S.W. 13., Eng.

Spaulding, M. H., A.m., '13 507 S. 8th Ave., Bozeman, Mont.

Sperry, Arthur B., B.S., '21 Kansas State Agr. College, Manhattan, Kansas.

Spurgeon, Charles H., A.M., '13 Sheridan, Ind.

Stewart, Thomas S., M.D., '17 18th and Spruce Sts., Philadelphia, Pa.

Stone, G. E Holly Drive, Los Angeles, Calif.

Stone, Grace A., A.M., '16 Teacher's College, New York City.

Stunkard, Horace W., Ph.D., '13 New York Univ., Univ. Heights, New York, N. Y.

Summers, Prof. H. E., '86 Ames, Iowa.

Swezy, Olive, Ph.D., '15 East Hall, University of Cahfornia, Berkeley, Cal.

Swingle, Prof. Leroy D., '06 Univ. of Utah, Salt Lake City, Utah.

Taylor, Sister Monica, M.S., '21 Notre Dame Training College, Glasgow, Scotland.

Taylor, W. R., Ph.D., '20 2223 W. Tioga St., Philadelphia, Pa.

Terrell, Truman C, M.D., '16 1301 Eighth St., Fort Worth, Tex.

Thomas, Arthur H., '99 12th and Walnut Sts., Philadelphia, Pa.

Timmins, George, '96 1410 E. Genesee St., Syracuse, N. Y.

TiNSLEY, Randolph Word, B.S., '15 Georgetown, Texas.

Titus, C. P., '22 32 New St., East Orange, New Jersey.

Todd, James C, B.A., M.D., '11 Boulder, Colo.

LIST OF MEMBERS 185

Tucker, O. C, Ph.G., '19 * 898 S. Clarkson St., Denver, Colo.

TuERS, R. v., '20 Dept. Biology, N. Y. Univ., New York, N. Y.

Van Cleave, Harley J., Ph.D., '11 300 N. H. Bldg., Urbana, 111.

Waite, Frederick C, Ph.D., '11

Medical Department, Western Reserve University, Cleveland, Ohio.

Walker, Elda R., Ph.D., '07 University of Nebraska, Lincoln, Neb.

Walker, Leva Belle, '13 Station A, Lincoln, Nebr.

Walton, A. S., '19 Island Heights, New Jersey.

Warbrick, J. C, M.D., '12 306 E. 43rd St., Chicago, 111.

Ward, Henry B., A.M., Ph.D., '87 University of Illinois, Urbana, 111.

Warren, S., A.B., '19 28 Hawthorne Rd., Brookline, Mass.

Waterworth, W. A., '15 286 Lambton Quay, W^ellington, N. Zealand,

Welch, paul S., Ph.D., '11 Univ. of Michigan, Ann Arbor, Mich.

Welsh, Major B. C, '14 24 Upper Mountain Ave., Montclair, N. J.

Weston, William H., Jr. Ph.D., '16

Herbarium & Lab. of Cryptogamic Botany, Harvard Univ., Cambridge, Mass.

Wheeler, E. J., Ph.D., '00 79 Chapel St., Albany, N. Y.

Whelpley, H. M., M.D., Ph.G., '09 2342 Albion PI., St. Louis, Mo.

Whiting, William J., '15 Optical Shop, Navy Yard, Washington, D. C.

Williamson, Wm., F.R.S.E., '07 79 Momingside Drive, Edinburg, Scotland.

Wilson, Charles Earl, A.M., '15

Dept. of Biology, Box 263, Wake Forest College, Wake Forest, N. Car.

Wilson, Ray W., '18 2051 Seneca, Buffalo, N. Y.

WiSMER, Nettie M., '19 Box 157, Erie, Kansas.

WoDSEDALEK, Jerry Edward, Ph.D., '15 Moscow, Idaho.

WoLCOTT, Robert Henry, A.M., M.D., '98 Univ. of Nebraska, Lincoln, Neb.

YosHiDA, Sadao, '20 Pathological Department, Osaka Medical College, Osaka, Japan.

Young, Paul A., A.B. '22 Dept. Botany, Univ. of Illinois, Urbana, 111.

Zeiss, Carl, c/o Dr. Boegehold, Dept. Biol Jena, Germany.

Zimmerman, Naomi B., M.Sc, '22 Kans. St. Agr. College, Manhattan, Kans.

Subscribers

Academy of Natural Sciences Logan Square, Philadelphia, Pa.

American Museum of Natural History. . .77th St. and Central Park, New York, N. Y.

Amherst College Library Amherst, Mass.

Babcock Scientific Library Plainfield, N. J.

Ball, Miss F. D Junior College, Grand Rapids, Mich.

BiBLioTHECA De La Facultad De Medicine Montevideo, Uruguay

Boston Public Library Boston, Mass.

Boston Society of Natural History Berkeley St., Boston, Mass.

Brown University Biological Library Prividence, R. I.

Bureau of Science Library Manila, P. I.

Carnegie Free Library Allegheny, Pa.

Carnegie Library Periodical Div., Schenley Park, Pittsburgh, Pa.

Chemists' Club Library, A. H. Elliott 52 East 41st St., New York, N. Y.

Chicago University Library Chicago, III.

Clarke, T. J 417 Third Ave., Brooklyn, N. Y.

Cleveland Public Library Cleveland, Ohio.

Coburn Library of Colorado College Colorado Springs, Colo.

Colby College Library Waterville, Me.

186 LIST OF MEMBERS

College of Physicians Library 19 S. 22nd St., Philadelphia, Pa.

College of William and Mary Library Williamsburg, Va.

Colorado Agricultitral College Library Fort Collins, Colo.

Colorado State Normal Library Greeley, Colo.

Columbia University Library , New York, N. Y.

Cornish Brothers 39 New Street, Birmingham, Eng.

De Pauw Univ., Alfred Dickey Biol. Library Greencastle, Ind.

Detroit Public Library Detroit, Mich.

Doane College Library Crete, Nebraska.

Drake University Library Des Moines, Iowa.

Dxjlau & Co 34-36 Margaret St., Cavendish Square, London, England.

Earlham College Library Earlham P.O., Richmond, Indiana.

Fargo College Library Fargo, N. D.

Fordham University Library Fordham, N. Y.

Franklin & Marshall College Library Lancaster, Pa.

George Washington University Library Washington, D. C.

Grosvernor Library Buffalo, N. Y.

Hamaker, J. J Randolph-Macon Woman's College, Lynchburg, Va.

Illinois Nat. Hist. Survey Library Urbana, 111.

Illinois Woman's College Library Jacksonville, 111.

Indiana State Library Indianapolis, Ind.

Indiana University Library Bloomington, Ind.

Institiito Oswaldo Cruz (chez M. Albanel) 11 Rue Sanbuier, Paris, France.

Iowa State Teachers' College Library Cedar Falls, Iowa.

James Millikin University Library Decatur, 111.

John Crerar Library Chicago, 111.

Johns Hopkins Univ. Library Baltimore, Md.

Kansas City Public Library Kansas City, Mo.

Kansas State Agr'l. College Library Manhattan, Kans.

Knox College Library Galesburg, 111.

Leland Stanford Jr., Univ. Library Stanford, California.

Library, Office of Surgeon General, U. S. Army Washington, D. C.

Los Angeles Public Library Los Angeles, Cal.

Martin, Mrs. Norman A 314 S. D. & T. Bldg., New Castle, Pa.

Mass. Agricultural College Library Amherst, Mass.

McGill University Library Montreal, Can.

Mendel Club Spring Hill College, Mobile, Ala.

Michigan State Normal College Library Ypsilanti, Mich.

Milwaukee Public Library Milwaukee, Wis.

Minnesota Univ., Farm Library St. Paul, Minn.

Missouri Botanical Garden St. Louis, Mo.

Missouri Valley College Library Marshall, Mo.

Montana State College of Agriculture Library Bozeman, Mont.

Mount Holyoke College Library South Hadlcy, Mass.

Museum Comparative Zoology (Harvard) Cambridge, Mass.

Museum of History, Science and .Art Exposition Park, Los Angeles, Calif.

Muskingum College Library New Concord, Ohio.

National Southeastern University Library Nanking, China.

New Hampshire State Library Concord, N. H.

New York Academy of Medicine 17 W. Forty-third St., New York City.

New York Microscopical Society 463 West St., New York City.

LIST OF MEMBERS 187

New York Public Library 476 Fifth Ave., New York City.

New York State Library Serial Section, Albany, N. Y.

Northwestern College Library Naperville, 111.

Oberlin College Library Oberlin, Ohio.

Ohio State University Library Columbus, Ohio.

Ohio Wesleyan University Library Delaware, Ohio.

Oregon Agricultural College Library Corvallis, Oregon.

Otteroom, Dr. Andrew Box 1126, Fargo Clinic, Fargo, N. Dak.

Pammel, Prof. L. H Department of Botany, Iowa State College, Ames, Iowa.

Peking Union Medical College Library Peking, China

Preside della Facolta Medica R. Universita di Pavia, Pavia, Italy.

Princeton University Library Princeton, N. J.

Puget Sound Biol. Station Library University of Washington, Seattle, Wash.

Purdue University Library Lafayette, Ind.

Queen's University Library Kingston, Ontario.

Rice Institute Library Houston, Texas.

Rutgers College Library New Brunswick, N. J.

ScRipps Institution Library La JoUa, Cal.

Smith College Library Northampton, Mass.

St. Mary's College Library St. Marys, Kansas.

Tennessee Agr. Experiment Station Knoxville, Tenn.

Texas Christian University Library Fort Worth, Tex.

Trinity College Library Durham, N. C.

Trinity College Library Washington, D. C.

U. S. Dept. "OF Agriculture Library Washington, D. C.

University of Arizona Library Tucson, Ariz.

University of Ark. Medical Dept. Library Little Rock, Ark.

University of California Library Berkeley, Calif.

Unr'ersity of Iowa Library Iowa City, Iowa.

University of Kansas Library Lawrence, Kans.

University of Michigan Library Ann Arbor, Mich.

University of Minnesota Library .Minneapolis, Minn.

University of Missouri Library Columbia, Mo.

University of Montana Library Missoula, Mont.

University of Nebraska Library Lincoln, Nebr.

University of Oklahoma Library Norman, Okla.

University of Oregon Library Eugene, Oregon.

University of Pennsylvania Library Philadelphia, Pa.

University of Southern California Library Los Angeles, Cal.

University of Texas Library Austin, Texas.

University of Toronto Library Toronto, Canada.

University of Utah Library Salt Lake City, Utah.

University of Virginia Library Charlottesville, Virginia.

University of Washington Library Seattle, Wash.

University of Wisconsin Library Madison, Wis.

University of Wyoming Library Laramie, Wyo.

Vassar College Library Poughkeepsie, N. Y.

Washington and Lee Biological Dept. Library Lexington, Va.

Washington State College Library Pullman, Wash.

Western College for Women Library Oxford, Ohio.

Yale College Library New Haven, Conn.

I

i

INDEX TO VOLUME XLI

Abnormal Earthworm Specimens, Helod-

rilus subrubicundus and H. tenuis, 153.

Absolute Ethyl Alcohol, Substitutes for, 155.

Alcohol, Substitutes for Absolute Ethyl, 155.

American Microscopical Society, Proceed-

ings of, 57.

Anatomy of Some Sexually Mature Speci-

mens of Dero limosa Leidy, 159.

Annual Report of the Treasurer, 110.

Aquatic Lepidoptera, Respiratory- Mechan-

ism in, 29.

Aspidogaster conchicola, Notes on the

Excretory System in, 113.

B

Barber Pipette, A Modified, 55.

C

Cestode from Liparis liparis, A New, 118.

Cleaning Slides and Covers for Dark-field

Work, 56.

Covers for Dark-field Work, Cleaning Slides,

56.

Cunningham, Bert. ]\Iodified Barber Pi-

pette, 55.

Custodian's Report for the Year 1921, HI.

D

Dark-field Work, Cleaning Slides and

Covers for, 56.

Dero limosa Leidy, The Anatomy of Some

Se.xually Mature Specimens of, 159.

Dichromatic Illumination for the IMicro-

scope, 51.

Douglas Lake, Michigan, Preliminary' Note

on Naids of, 167.

Earthworm Specimens, Helodrilus subrubi-

cundus and H. tenuis. Abnormal, 153.

Ethyl Alcohol, Substitutes for .\bsolute, 155.

Excessive Sexual Development in Hydra

oligactis with spermar>- on Tentacle, 172.

Excretory System in Aspidogaster conchi-

cola, Notes on, 113.

Faust, E. C, Notes on the Excretory System

in Aspidogaster conchicola, 113.

Frogs, On the Protozoa Parasitic in, 59.

Gage, S. H., Cleaning Slides and Covers for

Dark-field Work, 56.

Gregarines described from 1911 to 1920, A

List of the New, 122.

Griffin, L. E., Substitutes for Absolute

Ethyl .-Vlcohol, 155.

H

Hausman, L. A., Dichromatic Illumination

for the Microscope, 51.

Hayden, H. E., Preliminary Note on Naids

of Douglas Lake, Michigan, 167.

Heald, F. D., Some Suggestions for Teaching

Mycolog>s 175.

Helodrilus subrubicundus and H. tenuis.

Abnormal Earthworm Specimens, 153.

Helodrilus tenuis, .Abnormal Earthworm

Specimens, Helodrilus subrubicundus and,

153.

Heredity, Ten Years of, 82.

Hydra oligactis with Spermary on Tentacle,

Excessive Sexual Development in, 172.

I

Illumination for the Microscope, Dichro-

matic, 51.

Index, 189.

K

Kamm, Minnie Watson, List of the New

Gregarines described from 1911 to 1920,

122.

Killing, Staining and Mounting Parasitic

Nematodes, 103.

Kudo, R., On the Protozoa Parasitic in

Frogs, 59.

L

Lepidoptera, Respiratory Mechanism in

Certain Aquatic, 29.

Linton, Edwin, A New Cestode from Liparis

liparis, 118.

189

190

INDEX TO VOLUME XLI

Liparis liparis, A New Cestode from, 118.

List of members and subscribers, 179.

List of the New Gregarihes described from

1911 to 1920, .\, 122.

IM

Mature Specimens of Dero limosa Leidy,

The Anatomy of Some Sexually, 159.

May, H. G., Killing, Staining and Mounting

Parasitic Nematodes, 103.

Mayhew, R. L., The Anatomy of Some Sexu-

ally Mature Specimens of Dero limosa

Leidy, 159.

Members, List of, 179.

Michigan, Preliminary Note on Naids of

Douglas Lake, 167.

Microscope, Dichromatic Illumination, 51.

Micro-slip, A New, 101.

Mounting Parasitic Nematodes, Killing,

Staining and, 103.

Mycology, Some Suggestions for Teaching,

175.

Myers, F. L., A New Micro-slip, 101.

N

Naids of Douglas Lake, Michigan, Prelimi-

nary Note on, 167.

Nematodes, Killing, Staining and Mounting

Parasitic, 103.

New Cestode from Liparis liparis. A, 118.

New Gregarines described from 1911 to 1920,

1920, A, List, 122.

New Locality for Spongilla wagneri Potts,

A, 106.

Notes on the Excretory System in Aspido-

gaster conchicola, 113.

P

Parasitic in Frogs, On the Protozoa, 59.

Pipette, A Modified Barber, 55.

Preliminary Note on Naids of Douglas Lake,

Michigan, 167.

Proceedings of the .\mcrican IMicroscopical

Society, 57.

Protozoa Parasitic in Frogs, On the, 59.

R

Report for the year 1921, Custodian's, 111.

Report of the Treasurer, Annual, 110.

Respirator>' Mechanism in Certain .\f|ualic

Lepidoptera, 29.

Roberts, E. W., Some Interesting Studies on

Spider Anatomy, 107.

Root, F. M., A New Suctorian from Woods

Hole, 77.

S

Schmidt, A. W., Excessive Sexual Develop-

ment in Hydra oligactis with Spermary on

Tentacle, 172.

Sexual Development in Hydra oligactis with

Spermary on Tentacle, Excessive, 172.

Sexually Mature Specimens of Dero limosa

Leidy, The Anatomy of Some, 159.

Shull, A. Franklin, Ten Years of Heredity,

82.

Slides and Covers for Dark-field Work,

Cleaning, 56.

Specimens of Dero limosa Leidy, The Anat-

omy of Some Se.xually Mature, 159.

Smith, Frank, Abnormal Earthworm Speci-

mens, Helodrilus subrubicundus and H.

tenuis, 153.

Smith, Frank, a New Locality for Spongilla

wagneri Potts, 106.

Specimens, Helodrilus subrubicundus and

H. tenuis. Abnormal Earthworm, 153.

Spermary on Tentacle, Excessive Sexual

Development in Hydra oligactis with, 172.

Spider Anatomy, Some interesting Studies

on, 107.

Spongilla wagneri Potts, A New Locality for,

106.

Staining and Mounting Parasitic Nematodes,

KUlmg, 103.

Studies on Spider Anatomy, Some Interest-

ing, 107.

Subscribers, List of, 185.

Substitutes for Absolute Ethyl Alcohol, 155.

Suctorian from Woods Hole, A New, 77.

Suggestions for Teaching Mycology, 175.

Teaching Mycology, Some Suggestions for,

175.

Tentacle, Excessive Sexual Development in

Hydra oligactis with Spermary on, 172.

Treasurer, .Annual Report of the, 110.

W

Wagneri Potts, a New Locality for Spongilla,

106.

Welch, P.S., Respiratory Mechanism in

Certain .Aquatic Lepidoptera, 29.

Woods Hole, .A New Suctorian from, 77.

A,

TRANSACTIONS

OF THE

American

Miscroscopical Society

Organized 1878 Incorporated 1891

PUBLISHED QUARTERLY

BY THE SOCIETY

EDITED BY THE SECRETARY

PAUL S. WELCH

ANN ARBOR, MICHIGAN

VOLUME XLII

Number One

Entered as Second-class Matter August 13, 1918, at the Post-office at Menasha,

Wisconsin, under Act of March 3, 1879. Acceptance for mailing at the

special rate of postage provided for in Section 1103, of the

Act of October 3, 1917, authorized Oct. 21, 1918

(ilje (Tnllrginte 'J^rtsB

}L'CRGE BANTA I'UBLISHING COMPANY

MENASHA, WIS.

1923

TABLE OF CONTENTS

For Volume XLII, Number 1 , January, 1923

Studies on Sparganophilus eiseni Smith, with four plates, by Florence S. Hague 1

A Systematic Presentation of New Genera of Fungi, by O. A. Plunkett, P. A. Young,

and Ruth W. Ryan 43

Department of Methods, Reviews, Abstracts, and Briefer Articles

Hemistomum Confusum, a Homonym, by John E. Guberlet 68

The Use of Sodiumi Silicate as a [Mounting Medium, by Charles W. Crcaser and

William J. Clench .' 69

New Pocket Dissecting Microscope 71

The Physiology of Reproduction, a review b)- T. W. Galloway 72

Mar>' AUard Booth, by Bessie Perrault Titus 73

Proceedings of the American IMicroscopical Society 76

STUDIES ON SPARGANOPUILUS EISENI SMITH*

By Florence S. Hague

TABLE OF CONTENTS

I. Introduction 1

II. Specific Characters 3

III. Material and its Preparation 3

IV. Development 4

1 . Observ^ations on the Cocoons and Living Embryos 4

2. Embrj'olog}^ 6

V. ]Morpholog3' 7

L External Characters 7

2. Pharyngeal Glands 8

3. Digestive System 8

4. Vascular System 9

5. Excretory System 9

. (a) Structure of a Nephridium 9

(b) History of the Anterior Nephridia 12

Table 1 13

Table II 15

Table III 16

6. Reproductive Sj'stem 17

(a) Time of Development 17

(b) Genital Funnels and Ducts 18

(c) Sperm Sacs 20

(d) Spermathecae 21

(e) Accessory Reproductive Glands 21

Table IV 24

VI. Systematic Relations 32

VII. Summary 34

VIII. Literature Cited 34

I. Introduction

The genus Sparganophilus was established by Benham (1893) when

he described the first species, 5. tamesis. He found the specimens of this

species in one restricted area along the Thames River. Since they had not

been reported from other places in England, and since he was unable to

*Contributions from the Zoological Laboratory of the University of Ilhnois, No. 215.

1

I FLORENCE S. HAGUE

find them in other places along the River, he concluded that they had

been introduced there from some other part of the world. He thought it

possible that they had been brought with timber or plants from North

America, for America was considered the home of the most closely related

forms. In 1895, H. F. Moore reported the presence of this same species

in the banks of streams in the vicinity of Philadelphia. However, some of

the Philadelphia worms were examined by Michaelsen (1917) and, al-

though they were not adequately preserved for dissection, he found from

external characters that they were not S. tamesis. In response to a request

for material Doctor J. P. Moore, also of Philadelphia, wrote that he was

unable to find any of the worms in the spring of 1920, in places where they

had previously been found.

The next species reported was 5. eiseni. The specimens were found

in the Illinois River at Havana, Illinois, by Professor Frank Smith, and

described by him in 1895. .5'. eiseni has since been reported from Ohio,

Michigan, Indiana, and Florida. Eisen in 1896 described 5'. henhami from

Mexico; S. guatemalensis and S. carneus from Guatemala and Iowa,

respectively; 5. smithi and 5. sonomae both from California. The genera

which, according to Michaelsen (1917), are most closely related to Spar-

ganophilus are found in Central and South America.

This paper deals with certain organs and with variations in specimens

of S. eiseni from Florida, Iowa, Illinois, and Michigan, from which states

the genus has previously been reported, and also with specimens from

Louisiana and Wisconsin. Although certain points in embryology and

in the anatomy of several systems are given some attention, the sub-

sequent discussion is concerned chiefly with the excretory and repro-

ductive systems. The structure of a nephridium of S. eiseni is compared

with that of the nephridia of other genera which have been studied.

Although the development and subsequent loss of the nephridia of the

anterior somites has been known for some time among limicoline Oligo-

chaeta, a similar process has been reported in an earthworm only once.

The detailed developmental history of the nephridia of the anterior

somites of an earthworm is here presented for the first time. It has been

found that they develop and then disintegrate while the worms are still

quite young. The disintegration of the nephridia in the somites which

contain the genital ducts throws additional light on the relation of genital

ducts and nephridia. While accessory reproductive glands have been

reported in numerous species of earthworms and their structure described

in some of these species, the published accounts have been based on com-

paratively few specimens, and have not dealt with their development. The

data in the present paper have been taken from over 150 specimens which

were difTerent in age and which were collected at various seasons of the

year.

STUDIES ON SPARGANOPHILUS EISENI SMITH 6

The writer is indebted to Doctor C. P. Alexander of the State Labora-

tory of Natural History for the material collected at Havana, Illinois in

1920. Material collected in earlier years at Havana, and at Douglas Lake,

Michigan, as well as that from various other places, is in the collection of

Professor Frank Smith. The writer wishes to express her appreciation for

the use of this material and for other material which he has recently

collected. She is further indebted to Professor Smith for his suggestions

and interest during the progress of this study.

II. Specific Characters

Sparganophilus eiseni is a rather slender earthworm, which varies in

length from 80 mm. to 200 mm. Setae c and d are in the dorsal half of

the worm. The clitellum extends approximately from somites 15-25; the

tubercula pubertatis, from 17-22. The spermiducal pores are on somite

19, and the oviducal pores on 14, but both are inconspicuous; the sperma-

thecal pores are just ventrad of the seta line c, in the intersegmental

grooves 6/7, 7/8 and 8/9. There is no trace of gizzard or of calciferous

glands in the digestive tract. Moniliform hearts are present in somites

7-11. The spermathecae are in somites 7, 8 and 9; the ovaries, in 13; the

ovisacs, in 14; the spermaries and spermiducal funnels, in 10 and 11. The

sperm ducts pass through the longitudinal muscular layer of the body wall

in somites 11 and 12, and, from there to the pores, lie in the circular

muscular layer, or between it and the epidermis. The lobed sperm sacs

are paired and are in somites 11 and 12. The first typical nephridia are

in somite 13 or 15. Accessory reproductive glands may be present in some

one or more of somites 3 to 10, and are regularly present in 23-26 or adja-

cent somites. Both worms and cocoons are figured by Smith (1915).

III. Material and Its Preparation

Material for this study was collected between July 1919 and February

1921, in the region of Douglas Lake, Michigan, and at Homer Park and

Havana, Illinois. The collecting in the former locality was done while the

writer was in attendance at the University of Michigan Biological Station,

which is on the shore of Douglas Lake. Although search was made for

Sparganophilus at many points along the shores of Douglas Lake and of

its tributaries, specimens were found only in six places. These places are

near, and perhaps have been a part of the shore line at some past time, or

are now connected with the lake shore. They are supplied with decaying

organic matter but are separated from each other by stretches of shore,

parts of which are bare sand. Two mature worms were collected in one

spot on the northwest shore of the Lake, but since they were the only ones

in the vicinity, they may have been carried there from some other place.

Large numbers of worms were found in Diogenes Pond, at Hook Point, in

4 FLORENCE S. HAGUE

Sedge Pond, in the banks of Bessey Creek, and in the banks of Maple

River. Maple River flows out from Douglas Lake, and, after a long and

tortuous course, empties into Burt Lake which at the nearest point is less

than two miles from Douglas Lake. One specimen was brought in with

some plants collected by Doctor F. C. Gates at the mouth of this River

(Burt Lake). At the entrance of Carp Creek into Burt Lake, specimens of

Sparganophilus were abundant. Two unidentified lumbricids' were also

found here. This was the only place at which the writer collected another

species of earthworm with Sparganophilus eiseni. At Homer Park, Illi-

nois, specimens of S. eiseni have been collected in May, June, August,

September, October, December and February. They live along the bank

and in the bed of a small stream, Salt Fork, sometimes among gravel and

small stones, sometimes in soft, mucky soil.

In all these places in which worms were collected, decaying organic

matter was present. They were never more than 12 to 18 inches from the

edge of the water, and as the water gradually receded in Sedge Pond, the

fresh castings were always close to its edge. The worms were usually

found at a depth of one to four inches. Frequently both worms and cocoons

were found among the roots of grasses growing in or near the water.

The first step in preparing the worms for study was to clean out the

digestive tract. This was accomplished by keeping the worms in a vessel

with wet filter paper or cloth for 36 to 48 hours. They were then anes-

thetized with chloretone solution, straightened out between sticks and

killed. A solution of alcohol and formalin; a saturated solution of corro-

sive sublimate; and a solution of corrosive sublimate and acetic acid were

used as killing fluids. The latter was used exclusively for the smaller

embryos. Sections were stained with Ehrlich's haematoxylin, and a few

were counterstained with eosin or orange G.

IV. Development

(1) Observations on the Cocoons and Living Embryos

The cocoons of S. eiseni are elongated, but not all of them are as slender

and long as those figured by Smith (1915, figs. 8 and 9). The formation of

cocoons evidently occurs chiefly during July and August, and to a limited

extent during the last half of June and the first two weeks of September.

When first formed the cocoons are soft, colorless and transparent or

semi-transparent. They are enclosed in a slime tube which may be two

or three times as long as the cocoon. The cocoons gradually assume a

straw color, become less flexible, and lose the slime tube, probably within

six hours. They usually remain suflficicntly transparent so that the eggs

within them are visible. The cocoons which were formed after the worms

had been in the laboratory (in dishes with wet filter paper) for three or

STUDIES ON SPARGANOPHILUS EISENI SMITH 5

four days were misshapen and opaque in comparison with those which

were formed during the first day or two in the laboratory, or those which

were found in their natural environment. Although it is not uncommon to

find eight or ten eggs in a cocoon, the number of fairly well developed

embryos is usually one to four. Seven such embryos have been found in

one cocoon, and five or six in each of several cocoons. From a large number

of cocoons, perhaps 25% of those which were brought into the laboratory

and were apparently normal, worms did not emerge. Of those cocoons

from which worms failed to hatch, some remained perfectly transparent,

and some became more or less opaque. Many of the latter, when opened,

contained disintegrated embryos. In some of both transparent and

opaque cocoons, but chiefly the latter, ciliates were found. Some of these

cocoons were not tightly sealed, for a slight pressure caused them to emit

their contents. Ciliates from the surrounding water might have invaded

such cocoons. However, other cocoons were so tightly sealed that they

had to be cut open, and the ciliates were seen in the albumen as it was

forced out of the cocoon. In a few instances rhabdocoels, rotifers and

nematodes were apparently in the cocoons. Beddard (1892) reported

finding nematodes in the cocoons of Octockaetus (originally Acanthodrilus)

multiporus.

From two cocoons which were brought into the laboratory when the

eggs were in early cleavage or the blastula stage, worms hatched out in

24 and 26 days, respectively. Other cocoons were kept for 2>3 days before

the worms emerged. Empty cocoons are open only at one end. Some

of the worms, at hatching, are twice the size of others. Such a difi^erence

in size is found frequently, when there are two to six embryos in one cocoon.

Again one cocoon may contain eggs in cleavage stages and also well formed

gastrulae. It scarcely seems that the difference in the time of fertilization

would be great enough to cause such a difference in size. Possibly there

are periods of cessation of development, or variations in the rate of metab-

olism.

Four bifid embryos have been taken from cocoons. Each has a single

anterior part and two posterior ends. Two of these embryos were the

complete contents of one cocoon and were removed before they had

attained the degree of development which usually occurs within the cocoon.

The third was noticed in a cocoon which had been in the laboratory 34

days. It was moving actively, but, although the opening at the end of

the cocoon was enlarged, it did not escape. After eight more days the

posterior ends began to disintegrate and it was removed and fixed. A

study of these embryos shows a type of dorsal union. A single anterior

digestive tract bifurcates into the two digestive tracts of the posterior

ends. On opposite sides of the single digestive tract and continuing directly

posteriad into each branch, is a nerve cord with four pairs of setae in the

6 FLORENCE S. HAGUE

normal relative position. The anterior portions contain from 7 to 20 or 25

somites, and equal from one-seventh to three-fifths of the total lengths.

One bifurcated worm which had probably been out of the cocoon only

a few days was found in debris which had been brought into the laboratory.

An unsuccessful attempt to keep it alive resulted in the disintegration and

loss of the bifurcated portion. Figure 1 shows the manner of bifurcation,

which was posterior and which extended through about one-fifth of the

total length. The branch was somewhat shorter than the end which was

in line with the main axis, but, like it, had an anus, a growing region, a

ventral vessel and a pulsating dorsal vessel.

(2) Embryology

The study of the embryology of Sparganophilus eiseni is incomplete,

but because of the difiference of opinion in regard to several points in the

embryology of earthworms, it seems best to mention the more important

facts noted. Wilson described the following parts in the germ bands of

Lumbricus: a thin outer layer of ectoderm; a middle layer formed from

four pairs of large cells which are known as teloblasts and which are ecto-

dermal in origin; and an inner layer of mesoderm formed from large cells

which are known as mesoblasts. Two large mesoblasts with the meso-

dermal bands extending forward from them are present in embryos of

Sparganophilus eiseni. The teloblasts are somewhat anterior to the

mesoblasts and are less readily identified. In sections of 0.8 mm. and

1 mm. embryos they form a part of the surface ectoderm, but in 2 mm.

embryos they are more or less sunken beneath the ectoderm.

Continuous with and anterior to each teloblast is a row of cells to which

it has given rise. Because of the structures which develop from these

rows the median or first pair of rows are known as the neural rows, and

the second as the nephridial. Staff attributed the formation of the circu-

lar muscular layer of the body wall to the third and fourth rows. Earlier

investigators (Wilson, Bourne 1894a) had not reached the same conclusion.

In longitudinal sections of the 0.8 mm. embryo of 5. eiseni, the neural rows

are covered by a thin cellular layer and can be traced to the sides of the

mouth, but the second and third rows are at the surface and can be traced

forward through only a comparatively few sections. Even in sections

of a 2 mm. embryo the third and fourth rows are lost at times. In whole

mounts of 2 mm. to 3 mm. embryos the four pairs of rows show definitely.

The first pair are closely approximated in the midventral line; the second

row shows the heavy masses or coils of the early nephridia; the third row

is a straight line in which no structural differentiation shows; and the

fourth row is a series of oval bodies, one in each segment. The development

was not followed further, but the difference in the appearance of the third

STUDIES ON SPARGANOPHILUS EISENI SMITH 7

and fourth rows would scarcely indicate that they form the same adult

structure, namely, circular muscles.

V. Morphology

(1) External Characters

There is a wide variation in the size of the mature specimens of S.

eiseni collected. The largest are over 200 mm. in length and 2.6 mm. in

diameter, and the smallest are only 80 mm. to 100 mm. in length and

about 1 mm. in diameter. The former are from certain places in the

vicinity of Douglas Lake, Michigan, and the latter from Havana, Illinois.

In these places there are also worms of intermediate size. No regular

difference other than size has been noted among these worms.

The clitellum begins dorsally on somite 15 but may not include all of

that somite. The posterior end of the clitellum is somewhat variable.

There is usually a gradual decrease in thickness, beginning on 24 and

sometimes extending onto 26. Ventrally the clitellum is thin, and scarcely

distinguishable except in sections. This ventral part of the clitellum

extends from 14-26 or 27, and is usually somewhat thicker in the "region

of 22 and 26. On worms, which were killed in the manner described, the

clitellar region is definitely enlarged; but in some specimens which were

apparently put directly into formalin or alcohol, it is no greater in diameter

than the somites adjacent to it. It is, however, distinct because of a

slightly darker color and because of the absence of intersegmental grooves.

The tubercula pubertatis seem to be typically on somites 17-22, but

may extend onto 16 and 23. A specimen from Wisconsin has the tubercula

pubertatis on 18-22, and two from Homer Park, Illinois, in which the

clitella and tubercula pubertatis are developing, have the latter on somites

18-22.

Ventrally the clitellar somites are flattened and the tubercula puber-

tatis form longitudinal ridges along the lateral edges of part of the flattened

area. The flattened area usually extends posteriad onto 26 or 27, which

extent is one or two somites posteriad of the dorsal clitellar thickening.

Sometimes a pair of narrow ridges extend posteriad from the tubercula

pubertatis, along the lateral edge of this flattened area and border its

rounded posterior end. The posterior end is always rounded and the

whole flattened area definitely outlined, when the clitellum is well devel-

oped, even if the ridges are not present. Narrow ridges also extend

anteriad from the tubercula pubertatis in some specimens. The tubercula

pubertatis do infringe on the width of the flattened area between them, but

in neither the complete ventral flattening, nor the anterior part of it,

has the writer noted in any specimen the hour-glass shape which was

mentioned by Eisen (1896) as occurring in S. eiseni.

8 FLORENCE S. HAGUE

(2) Pharyngeal glands

The term, pharyngeal glands, is used in this discussion for those masses

of deeply staining cells, which are associated with the muscles extending

from the pharynx to the body wall of 5. eiseni. In somites 5, 6 and 7

the cells are aggregated in masses which are attached to the large pharyn-

geal muscles extending through those somites. The septum 3/4 is incom-

plete and the cells of this region are scattered singly and in small groups

between the muscles, near their attachment to the pharynx. Eisen (1896)

called the latter salivary or suprapharyngeal glands, and the former

septal or intestinal glands. He described ducts, which open into the

pharynx, from both kinds of glands. The writer has not found such ducts

in S. eiseni. Stephenson (1917) was unable to find any ducts from similar

glands in several species of Pheretima and of Helodrilus. He concluded,

f ro;n his studies, that both pharyngeal and septal glands were of peritoneal

origin, and were not related to the pharynx in the manner originally

supposed. The name, chromophil cells, was suggested by him for these

structures. It seems to the writer that it would be best to retain the

name, pharyngeal, until the origin of these cells from some source other

than the pharyngeal wall is definitely established. The term gland is used

for convenience.

Eisen stated that the glands of somite 6 were as large as those of 5 in

some species, but smaller in others. Table IV (p. 24) shows that the

relative size of the glands of 6 is variable, and that glands are sometimes

present in somites 7 and 8. Except in the immature specimens, the glands

of 6 are usually somewhat smaller than those of 5, in worms which were

collected in Michigan. In some of the specimens collected at Homer

Park, Illinois, the glands in 6 are as large as those in 5, and in others,

they are smaller. The writer could find no fixed relation between the

presence of the anterior accessory reproductive glands and the size of the

pharyngeal glands of 6.

It has been suggested that such pharyngeal glands are homologues of

the nephridia in certain worms in which the latter are absent in the anterior

somites. The subsequent discussion of the development and disintegration

of the anterior nephridia (p. 12) and the theory (Hesse) of the origin of the

pharyngeal glands from (he pharyngeal wall would both lend to contradict

the suggestion.

(3) Digestive System

The digestive tract has neither gizzard, esophageal glands, nor typhlo-

sole. A dorsal sac opens into the pharynx and the inner surface of the

esophagus usually has numerous irregular, but cliiclly transverse folds

in somites 4 and 5. In the succeeding somites it is folded longitudinally.

In 9 or 10 and one or more succeeding somites the diameter of the digestive

STUDIES ON SPARGANOPHILUS EISENI SMITH 9

canal is frequently greater, and the thickness of the walls is less than in

the somites immediately anteriad or posteriad. These facts indicate that

the enlargement is a temporary expansion. Since the perienteric blood

sinus begins in somite 9, the intestine may be said to begin in 9, although

no other structural difference has been found by which to distinguish

esophagus and intestine.

(4) Vascular System

The main vessels of the vascular system of S. tamesis and S. eiseni are

similar. They have been described by Benham (1893) and Smith (1895),

respectively. Eisen (1896) added the description of the blood glands,

but he was in error in stating that the hearts are in somites 8 to 11 in 6*.

eiseni. In all specimens studied, including one identified by Eisen as

.S". henhami, the hearts are in somites 7-11. The hearts decrease in size

from posterior to anterior, but all five pairs can be seen pulsating in living

worms. The hearts do not contract simultaneously, but quickly and in

rapid succession beginning with the most posterior one. It is evident that

the condition of the hearts at the time when the worms were killed (whether

they had just contracted, or were fully expanded) would make a difference

in their size. Among the worms studied, the hearts are all moniliform,

but in some they are more contorted and of relatively greater diameter.

(5) Excretory System

(a) Structure of a Nephridium

The excretory organs of S. eiseni are paired meganephridia which

open to the exterior through pores placed anteriad of the ventral setae.

These nephridia are large, compact organs, consisting of a lobed mass of

coelomic epithelium in which the nephric tubule is embedded. Similar

nephridia have been described in other species of Sparganophilus and in

other genera. They present an appearance quite different from that of

the nephridia of Lumbricus (Benham 1891), of M aoridrihis rosae (Camer-

on) or of the widely distributed Helodrilus caliginosus trapezoides. In

the latter a nephridium does not appear as a mass of tissue but as a series

of loops or a group of convoluted tubules. The tubule, however, is the

essential part in both types of nephridia.

The nephridia of Sparganophilus eiseni are so compact that tracing

the complete course of the tubule in the entire nephridium is impracti-

cable, if not impossible. Tracing the tubule in sectioned nephridia is

equally difhcult. Of several nephridia which were dissected out, stained

in Delafield's haematoxylin and cleared in glycerine, one was sufficiently

spread out to trace the greater part of the course of the tubule. The

nephridium was then sectioned, approximately in the plane outlined.

10 FLORENCE S. HAGUE

The single line in figure 2 represents the general course of the tubule as it

was worked out from the study of the whole mount and the sections.

The septum, nephrostome and duct connecting the latter with the nephrid-

ial mass were torn off, and are consequently shown by broken lines.

The break (br) was caused by displacing the lobe from its position over

the body of the nephridium. Aside from these interruptions the diagram

shows an unbranched and continuous tubule from the nephrostome to

the nephridiopore. Most of it is in two long and somewhat convoluted

loops.

Each nephridium, as is generally true, consists of a pre-septal and a

post-septal part. The former includes a nephrostome or funnel and a

short duct. The funnel is really a broad tube with two extensions, a large

lip and, opposite it, a very small lip. In the large lip the large marginal

cells and, outside of them, a row of extra-marginal cells are distinctly

visible from the front or inner side of the lip. When this lip is sectioned,

there is a noticeable thickening of the edge produced by one or two addi-

tional rows of extra-marginal cells, which curve over the edge onto the

outside of the lip. The smaller lip is merely a slight extension of the cells

of the tube and presents no special structures. Flattened coelomic epi-

thelium covers the outside of the entire funnel and continues over the duct.

The broad tube which forms the body of the funnel gradually narrows

into the duct which connects the nephrostome with the nephridial mass.

The cell walls are not visible, but judging by the number and position of

the nuclei, the tubular portion of the funnel is made up of two series of

cells. Just anterior to the septum the number of nuclei indicates that a

single row of perforated cells forms the duct. There is probably a gradual

change from the intercellular to the intracellular condition, as the funnel

narrows into the duct.

There is nothing to indicate that the post-septal part of the nephridium

is not intracellular throughout its length. It may be distinguished, accord-

ing to its structure, into three parts which are indicated in figure 2. They

are the narrow tube (nar) which forms the first loop; the middle tube

(mid) which extends to the apex of the second loop; and the wide tube

(wi) which extends from the apex of the second loop to the nephridiopore

(ne). The narrow tube is slender and thin-wallcd. Parts of it are ciliated.

The middle tube has a thicker wall and is ciliated throughout its length.

The first part of this tube is coarsely granular. Somewhat beyond the

middle the coarse granules gradually disappear and scattered groups of

fine granules appear. The fine granules increase in number and become

aggregated so that in the last of the middle tube they form a layer near the

middle of the wall of the tube (fig. 9). The granules are arranged rather

regularly in closely packed radial rows, with occasional strings of granules

extending out into the peripheral portion of the wall. In some ncphridia

STUDIES ON SPARGANOPHILUS EISENI SMITH 11

these strings are sufficiently numerous and branched to form a network.

This layer of granules stains differently from other parts of the nephridium

and is distinct in all nephridia.

A constriction separates the middle and wide tubes (fig. 9). The

wide tube is without cilia in all parts but the structure of the wall is not

the same throughout. A short portion, the ampullar region, at the begin-

ning of the wide tube is made up of rather large deeply stained and defi-

nitely outlined cells. Both cross and longitudinal sections of this portion

show very distinctly the intracellular nature of the nephric tubule. The

first third or half of this region is an enlargement, the ampulla (am),

which contains deeply staining masses in its lumen. The sectioned walls

show a darker inner and a lighter peripheral cytoplasm. In the peripheral

cytoplasm there is a network of granules which is similar to that in the

peripheral cytoplasm of the middle tube, and which seems to condense

into the darker inner cytoplasm. The inner cytoplasm differs from the

layer in the middle tube in that it lacks the radial striations and stains

less intensely. The distinction between inner and peripheral cytoplasm

gradually disappears in the large cells beyond the ampulla. These large

cells abruptly give place to a thin-walled tube in which the cell boundaries

are indistinguishable. The last part of the wide tube (fig. 5) begins with

cells similar to those of the ampullar region, but in its distal part the wall

becomes thinner and the cell boundaries gradually disappear. Figure 7

represents the distal part of the wide tube, including that which passes

through the body wall to the epidermal^ invagination, in a nephridium

which had been dissected out and sectioned. The number of nuclei in

this and adjacent sections of the tube indicates that it is intracellular.

Figure 8 shows a section of the complete epidermal invagination of the

same nephridium. Since adjacent sections show no cellular structure

other than that which is shown in these two figures, it is evident that the

wide tube opens directly into the nephridiopore, and that there is no

muscular duct. Serial sections of nephridia in situ substantiate this

conclusion.

The shape of the nephridium and position of the different parts of

the tubule within it are variable. However, the two long loops are regu-

larly approximately parallel, and for the greater part of their length, are

embedded in large lightly staining cells (fig. 6). These large cells are

usually found about the periphery rather than in the central part of the

nephridium, and they have not been noted except in relation to the nephric

tubule. These facts suggest that the large cells may have a direct relation

to the process of excretion.

^ The outer cellular layer covering the earthworm has been called epidermis, and also

hypodermis. Neither name is satisfactory, but because of its moie general usage the former

is used here.

12 FLORENCE S. HAGUE

The smaller, evenly granular cells, which form the nephridial mass,

are probably epithelial. The septa are covered by delicately stained

projections, some large, some small and closely crowded; some with nuclei,

others without nuclei. These projections are parts of vesicular epithelial

cells. Similar projections, without nuclei, cover the surface of the nephrid-

ium. These projections are frequently found to be parts of the evenly

granular cells of the nephridial mass. Evidently, then, these smaller

cells in the nephridial mass are epithelial cells, but only those at the surface

are vesicular.

The structure of the nephridial tubule of Lumbricus has been de-

scribed by Gegenbauer, Benham (1891) and Maziarski (1905). The last

paper is available only in the form of a review by Meisenheimer. K. C.

Schneider studied the nephridium of Eisenia rosea, one of the Lumbricidae,

and Cameron, the nephridium of Maoridrihis rosae, one of the Megas-

colecidae. The nephridium of each of these has three parts similar to

those of SparganophUus eiseni, and in addition a muscular duct which is

located between the wide tube and the pore. Intermittent ciliation of the

narrow tube, complete ciliation of the middle tube and lack of ciliation

in the wide tube are characters common to all. Histologically there are

differences in the structure of corresponding parts of the tubule. None,

except S. eiseni, have the more deeply staining layer in the wall of the

middle tube. The figure by Maziarski (Meisenheimer) shows some vari-

ation in the wide tube, but it is not the same as that found in S. eiseni.

The nephridia of Moniligastcr grandis (Bourne 1894b), and of Perieodrilus

ricardi and P. montanus (Benham and Cameron) are distinctly different

in structure from those of the above mentioned earthworms.

From the foregoing discussion it appears that although there is a

general agreement between the nephric tubule of SparganophUus eiseni

and that of Lumbricus, from the nephrostome to the muscular duct, the

former has a higher degree of specialization in the middle and wide tubes.

(b) History oj the A nterior Nephridia

Sparganophilus is one of several genera of earthworms which resemble

certain limicoline forms in lacking nephridia in the anterior somites.

Smith (1895) and Eisen (1896) both listed somite 13 as the first one contain-

ing nephridia in S. eiseni. In other species the first nephridia have been

recorded in 12, 13 and 16. However, a study of embryos shows that nephri-

dia are present in some of the somites anterior to 13 (Table I). The first

distinct ne])hridia are in somite 3 of seven embryos; in somite 4 of five em-

bryos; in somite 6 of four embryos and in somite 12 of two young worms.

Unfortunately no first nephridia have been found in somites 9, 10 and 11,

and only one in each of somites 7 and 8. In addition to the fact that,

generally in the larger embryos, the nci)hridia arc absent in the more

STUDIES ON SPARGANOPIIILTJS EISKNI SMITH

13

anterior somites, there are signs of disintegration of the anterior nephridia.

The first nephridia are called distinct rather than well developed or typi-

cal because, while there can be no doubt that they are nephridia, the canal

in some is indistinct if visible at all. This must be due to disintegration

for the canal develops very early as is shown by its presence in figure 11,

which is typical of the nephridia of three successive somites, probably 3, 4

and 5, of a 0.5mm. embryo. Figures 10 and 12, from sections through

nephridia of somite 3 of embryo No. 2 and of somite 5 of embryo No. 7,

respectively, show very distinct canals; but in figure 13 from a section

through one of the nephridia of somite 3 of embryo No. 7, only one indis-

tinct section of the canal is recognizable. The canal shows more definitely

in this section than in any other sections of the nephridia of this somite.

Table I

First Nephridia in Embryos and Very Young Worms

Nos. 20 and 21 are the same as 99a and 100a of Table IT, respectively.

Other first or first and second nephridia show similar conditions and in

somites anterior to the first nephridia there are frequently small masses of

tissue which are located and stained similarly to nephridia. These are

evidently rudiments of nephridia. They are in somite 3 of Nos. 9, 10, 11

and 12, and in one to three somites anterior to the first nephridia of Nos. 13

to 20. In somite 2 of several embryos there are small masses of tissue which

may be nephridial rudiments, but no nephridia, as distinct as the smallest

of those of somite 3, have been definitely located in somite 2. All these

facts indicate that the nephridia develop in the anterior somites and then

disintegrate in an antero-posterior order. If the number of somites rather

than the length of the embryo had been used as a measure of relative

development, there might be less variation among those worms which have

the first nephridia in 3, 4 or 6. Part of the variation in size is doubtless due

to the degree of contraction or relaxation of the embryos. However, such

14 FLORENCE S. HAGUE

a marked divergence as is shown by Nos. 7, 12 and 19 seems to indicate

variations in the rate of disintegration of the nephridia. Possibly such

variations are correlated with differences in the rate of metabolism, which

differences, as was suggested above, may be related to the distinct differ-

ences in size frequently found among embryos in the same cocoon.

The development of the nephridia takes place in an antero-posterior

direction. Each nephridium is consequently visibly farther advanced than

those from eight to ten somites posteriad of it. But, since the rate of devel-

opment is more rapid than that of disintegration, the nephridia of somites

10-12 reach a more advanced stage of development than do those of somites

3-6. In No. 14 the nephridia of somite 7 have more convolutions and a

canal of proportionately greater diameter than do the nephridia of somite

3. In somites 7 and 8 of No. 18, large cells, similar to those in which part

of the nephric tubule of the adult is embedded, can be recognized. In

No. 20 the epithelial masses have developed about the nephridia of 12.

The development of the nephrostome has not been followed, but there are

funnels in 9 and some of the succeeding somites of No. 19, and in several

somites of No. 18. The nephridiopores are readily recognizable in somite 8

of No. 19 and in somites 7-10 of No. 18, but not in other worms in which the

nephridia extend as far forward as somites 7 and 8. In specimens in which

typical nephridia are present in 12 the pores are distinct in that somite.

A study of mature and immature worms discloses a variation in the

position of the first typical nephridia. Table II shows the condition of

the nephridia in those worms in which they were studied. Table III is a

summary of Table II. A typical nephridium is one, such as has been de-

scribed, with its tubule embedded in a mass of cells, and having a definite

nephrostome and nephridiopore. The reduced nephridia vary from a small

mass of epithehal cells without a tubule to a mass about the size of a normal

nephridium, but with reduced tubule and without a nephrostome or a

nephridiopore. The typical and reduced nephridia, especially of 13,

grade into each other so that it is sometimes difficult to distinguish them.

This is because the disintegration of the nephrostomes is variable, and the

nephridiopores may persist and retain an indistinct connection with a

nephridium in which the tubule is reduced. Table III indicates that

reduced and typical nephridia are present in increasing numbers in somites

11, 12, 13 and 15, and that all nephridia in 15 are typical. All nephridia in

somites posteriad of 15 are also typical.

The increasing number of nephridia in somites 11, 12, 13 and 15 is

doubtless due to the antero-posterior order of disintegration. The dif-

ferent conditions of the nephridia of 12, of 13 or of 14 in different individ-

uals of S. eiseni are probably due to individual variations. It is possible

that the nephridia of these somites are reduced increasingly wilh age

or successive breeding seasons; Init, since no method has been found for

STUDIES ON SPARGANOPHILUS EISENI SMITH

15

distinguishing worms of different ages, there is no evidence for such a

progressive reduction. On the other hand, individual variations have

been noted in the nephridial condition of embryos. Specimen No. 101a

Table II

Nephrtdia of Young and Adult Worms

I indicates a typical organ: s, a reduced organ: o and a blank, absence of the organ.

Two symbols are used when the nephridia of the two sides are dissimilar. Identical specimens

have the same number as in Table IV.

Nephridia in somites

11

12

so

s

13

14

15

shows a marked difference from others of its size. Adult specimens

from different localities show greater or lesser tendencies toward the dis-

appearance of nephridia in certain somites.

16

FLORENCE S. HAGUE

Other worms in which the most anterior nephridia are in some one of

somites 10 to 16 are Sparganophilus tamesis, Criodrilus lacuum, AUuroides

pordageijGlflssoscolex giganteus (originally Titanus hrasiliensis), Haplotaxis

heterogyne and several species of Pontodrilus. The embryonic condi-

tion of the anterior nephridia of these is not known. Vejdovsky (1888-

1892) described the development and subsequent disintegration, during

embryonic life, of the nephridia of 1-6 of Rhynchelmis, and mentioned a

similar process in Chaetogaster, Aeolosoma and Nais. Bourne (1894a)

found that the nephridia of Diporochaeta (originally Perichaeta) pellucida

attained a well developed condition in somites 2-6 and then degenerated,

while those of 7-11 became complex. All of the worms mentioned, which

lack nephridia in ten or more anterior somites, live in or at the edge of the

water.

Bage and Stephenson (1915) found that, in certain Megascolecidae,

micronephridia are present in all somites of the body, and that in the pos-

terior somites, sometimes as far forward as 12, there are also meganephridia.

Table III

Summary of Table II

All with nephridia Is or to are counted with /.

All with nephridia so are counted with s.

Still other species of earthworms have a thick ccelomic covering on the

posterior but not on the anterior nephridia, or the muscular duct may be

more highly developed in one part of the body than in another. In a few

species the nephridia arc larger in a few anterior somites than in the poste-

rior ones. Such modifications are similar to the absence of nephridia in

the anterior somites.

1

STUDIES ON SPARGANOPHILUS EISENI SMITH 17

It has been thought that the absence of anterior nephridia in earth-

worms is correlated in some way with the reproductive system. However,

disintegration of the nephridia of Sparganophilus eiseni begins before there

is any trace of the reproductive organs. The gonads, which are the first

parts of the reproductive system to develop (p. 17), are not recognizable

until the nephridia of somite 8 are beginning to disintegrate (No. 19).

Furthermore, if the nephridia disintegrated as the reproductive organs

developed, they would disappear not in an antero-posterior sequence, but

first in somites 10, 11, 13; then in 11, 12, 14; lastly in 7, 8, 9; and probably

not at all in the somites anteriad of 7, since there are no reproductive

organs in those somites. The probable influence of the genital ducts on

the nephridia of 11, 12 and 14 will receive attention in the discussion of

the genital ducts. The time and order of the disintegration of the nephridia

of 3-10 indicate that this process is not directly related to the development

of the reproductive organs. The adaptation to the aquatic habitat or

some other factor may have produced a physiological condition which is

different from that in most earthworms and which has resulted in the loss

of the anterior nephridia.

(6) Reproductive System

(a) Time of Development

The development of the reproductive organs begins about the time

that the worms emerge from the cocoons, and is not closely related to the

size of the worms. The gonads appear first; then the spermiducal funnels,

sperm ducts and sperm sacs develop in quick succession. The oviducal

funnels, oviducts and ovisacs develop later, only a short time before the

accessory glands and spermathecae which develop as the worm approaches

adult size. Finally, with the approach of the breeding season, the clitellum

appears. Specimen No. 19 of Table I is the youngest specimen in which

the gonads can be identified, and in that only on one side. Nos. 20 and 21

are the same as Nos. 99a and 100a of Table II, respectively. These two

and the other immature worms, Nos. 99 to 102, inclusive, (Table II) have

distinct spermaries and ovaries. Nos. 99, 101a, 101 and 102 have anlagen

of the spermiducal funnels, and the latter three, of the sperm sacs and

sperm ducts also. Tiny anlagen probably of the oviducal funnels, are

present in Nos. 101 and 102, and definite anlagen of these funnels are

present in a 55mm. worm which is not included in the tables. In Nos. 121

and 122, which have reached adult size but probably not sexual maturity,

the oviducal funnels are still small and the anlagen of the oviducts are

recognizable. Accessory reproductive glands and spermathecae are not

developing in any of these specimens except Nos. 121 and 122.

18 FLORENCE S. HAGUE

Bergh (1886) found that the spermaries and ovaries were the only

parts of the reproductive system of Lumbricus, which developed during

embryonic life. In Octochaetus (originally Acanthodrilus) nmltiporus,

Beddard (1892) found, in addition to the gonads, prominent genital funnels

in somites 10-13 of embryos that were not yet ready to em.erge from the

cocoon.

(b) Genital Funnels and Ducts

The anlagen of the genital funnels first appear as deeply staining areas

on the anterior faces of septa 10/11, 11/12 and 13/14, just laterad of the

nerve cord and near the point at which the nephridial tube penetrates the

septum. This area thickens into a mass with deep indentations, from

which a deeply staining strand of tissue can be traced through the septum,

and toward or into the body wall of the following somite at the point where

the nephridium (if present) penetrates to the exterior. This strand is the

anlage of the genital duct. It enlarges and as sexual maturity approaches,

acquires a lumen. The oviduct opens directly to the exterior in 14, but the

sperm ducts pass into the body wall in 11 and 12 and then turn posteriad;

those of each side unite and extend, in the body wall, to the spermi-

ducal pores on somite 19. An early stage in the development of the longi-

tudinal ducts has been found in a worm in which the genital funnels are not

of maximum size, and the spermathecae and accessory glands are very

small. In longitudinal sections the duct appears as two narrow parallel

bands in the longitudinal muscular layer. In each band are many nuclei,

while numerous similar nuclei are located close to the duct and scattered

through the longitudinal muscular layer. Since the normal position of the

longitudinal part of the sperm duct in the adult is between the epidermis

and the circular muscular layer or in the latter, there must be a shifting of

this duct as it develops.

No difference has been noted in the development of the spermiducal

and oviducal funnels or in the ducts from them to the body wall. The

mature spermiducal funnel is much larger than the oviducal funnel, but

sections show that the difference is one of size rather than structure. Be-

tween periods of breeding activity the funnels revert to small solid masses

and the ducts to soHd strands of tissue.

Although the nephridia degenerate more or less completely before the

genital funnels develop, there is a limited amount of evidence in regard to

the morphological relations of excretory and reproductive systems. The

genital funnel anlagen first appear as parts of the epithelium of the septa,

while the nephridial funnels, at all stages seen, arc separated from the

septa by the short prc-septal portions of the ducts. In Nos. 101 and 102,

the nephridial funnels are still typical in somite 13, and there are also small

deeply stained areas which are pr()bal)ly the anlagen of the oviducal fun-

STUDIES ON SPARGANOPHILUS EISENI SMITH 19

nels. On one side of somite 13 of No. 122 there is a small mass of tissue,

which has the shape of a nephridial funnel, and which is in line with the

nephridial funnels of 12 and 14. It is probably the remnant of the nephrid-

ial funnel, and laterad of it is the very definite anlage of the oviducal

funnel. From the latter, the anlage of the oviduct can be traced through

the septum and toward the body wall. These facts indicate that the geni-

tal funnels are not derived from the nephridial funnels. The development

of the genital ducts from the genital funnels has been described above.

The oviduct passes into the body wall close beside the nephridium and

opens to the exterior through an epidermal invagination, which, if one

may judge by its position, is the nephridiopore. The sperm ducts likewise

pass into the body wall close beside the nephridia, but they open to the

exterior on somite 19, and entirely independent of the nephridia.

Vejdovsky (1884) described the anlagen of the genital funnels as

thickenings of the septa in Naididae, Enchytraeidae, Tubificidae and

Lumbriculidae. Bergh (1886) found the same type of development in

Lumbricidae. In Tubifex riviilorum, Gatenby found a difference in the

development of spermiducal and oviducal funnels, but both developed

from coelomic epithelium. All these investigators found that the genital

ducts developed as outgrowths from the genital funnels.

Stole (Beddard 1892) reported that in Aeolosoma the spermatozoa

passed out by way of the nephridia, especially those of somite 6, and that

during the sexual period the nephridia of certain somites disappeared

wholly or in part. In Marionina sp. (originally Enchytraeoides marioni)

Roule found that the nephridia in 1-8, and in 12 were lacking in early

stages of embryos; that the sperm ducts subsequently developed in 12,

and that with the attainment of sexual maturity the remaining nephridia

of 9-14 disappeared more or less completely. He regarded the sperm ducts

as nephridia whose development had been delayed. It seems to the

writer more probably that the nephridia of 1-8 and of 12 of Marionina had

developed and disintegrated in earlier stages, and that the sperm duct

probably originated as Vejdovsky described it in Enchytraeidae.

Beddard (1892) concluded that the genital funnels developed from the

nephridial funnels, and the genital ducts, from parts of the nephridial

ducts in Octochaetus multiporiis. The conditions in this species are similar

to those in Sparganophilus eiseni, except that in the latter there is no

apparent continuity between the genital and nephridial ducts. Benham

(1904) found, in both mature and immature specimens of Haplotaxis

heterogyne, that the most anterior nephridia were in somite 10 and were

somewhat degenerate, and that there were no nephridia in somites 11-13.

Because of the marked structural similarity between sperm ducts (in 11 and

12) and nephridia, he concluded that the sperm ducts were nephridia onto

which genital funnels had been grafted. Since the oviducts had no resem-

20 FLORENCE S. HAGUE

blancc to the nephridia, they were considered different in origin. No

explanation of the absence of nephridia in 13 was offered.

Reduction of the nephridia in the genital somites alone was reported

in Moniligaster (Benham 1886) and in Gordiodrilus (Beddard 1894).

In Libyodrilus violaceous Beddard (1891) found nephridia in all somites

posterior to 3 in a young specimen; but in the adult all that remained of

the nephridia of 13-17 was the integumental network. Beddard attributed

the loss of nephridia to the presence of very large spermathecae which

extended through these somites.

It has been concluded (p. 17) that the disintegration of the nephridia

of somites 3-10 of Sparganophilus etseni is not directly related to the devel-

opment of the reproductive organs; and (p. 19) that the genital funnels

and ducts develop independently of the nephridia, although the same

pore may serve first for a nephridium and subsequently for an oviduct.

With the exception of definitely immature specimens, none have typical

nephridia in 11, 12 and 14, in which somites the genital ducts are located.

The nephridia of 11 are merely rudiments or are lacking in those specimens

in which the anlagen of the spermiducal funnels are found. Of the imma-

ture specimens, Nos. 99a, 100a, and 100 have no spermiducal funnels but

do have typical nephridia in 12; Nos. 99, 101, 101a and 102 have the

anlagen of spermiducal funnels but have reduced or no nephridia in 12.

Nos. 101 and 102 have tiny anlagen of the oviducal funnels and typical

nephridia in 14; Nos. 121 and 122 have distinct anlagen of the oviducal

funnels and ducts, but the nephridia are reduced. Finally, 45 mature

worms have typical nephridia in 13 but reduced or no nephridia in 14

(Table III). Because of these facts, it seems that the disintegration of

anterior nephridia which is produced in the embryo by the physiological

condition, is augmented in 11, 12 and 14 by the development of the geni-

tal funnels and ducts of the respective somites. Since the nephridia are

not transformed into genital organs, probably this effect is produced by

introducing some different physiological condition, possibly absorption of

the food supply or a hormone secretion.

(c) Sperm Sacs

The sperm sacs extend into somites 11 and 12 from septa 10/11 and

11/12. During the breeding season, the sacs of 12 are often so large that

they push the septum 12/13 back against septum 13/14. Small out-

growths which somewhat resemble the sperm sacs are frequently found on

septum 12/13. The sperm sacs have been described as lobulate. Eisen

(1896) distinguished between a minutely lobulate condition in .S". eiseni

and a less minutely lobulate condition in S. bctihami. There is a great

variation in the size of the k)bules in S. eiseni at different seasons. When

free from spermatozoa the lobules are quite small, but at the height of

STUDIES ON SPARGANOPHILUS EISENI SMITH 21

the breeding season, they may be so distended and the whole sperm sac

so compact that the lobulation is not readily recognizable. Figures 99 and

119D (Eisen 1896) of S. benhami represent conditions frequently found

m S. eiseni.

(d) Spermathecae

Between periods of breeding activity a spermatheca is represented by

a small mass of undifferentiated cells. The mass is without a definite

lumen, and is placed at the inner end of a more deeply staining strand of

tissue which extends through the body wall. The strand is the rudiment

of the spermathecal duct.

The spermathecae of the sexually mature worm are variable in shape.

Often they are cylindrical with a spherical enlargement at the free end;

but the enlargement may be elongated at the expense of the cylindrical

portion. Sometimes the contour is smooth, again it is irregular as if the

spermatheca had been crowded into a space shorter than the spermatheca

itself. Eisen (1896) figured spermathecae of S. benhami (figs. 118a and

118b), of S. carneus (figs. 140a and 140b), and of S. guatemalensis (figs.

141a, 141b and 141c). A specimen, identified by Eisen as .S. benhami,

is in the collection of Professor Smith. It has spermathecae which are

slightly irregular in outline, but not so irregular as those figured by Eisen.

Some specimens of S. eiseni, which have the anterior accessory glands,

have spermathecae of the same shape as those of the above mentioned

specimen of S. benhami. Figures 140a, 140b and 141b (Eisen) are quite

typical of the shape of the spermathecae of S. eiseni. A sac, similar to the

ones which he mentions and figures at the end of the spermathecae of

S. guatemalensis, has been noted in some specimens of S. eiseni.

(e) Accessory Reproductive Glands

These are glands which are evidently related to the reproductive

activities but are not connected with the sperm ducts or other reproductive

organs. There are two kinds, of which one is found in some of somites

3 to 10, and the other in some of somites 15 to 17 and 22 to 27. For the

sake of brevity, the first mentioned glands will be called the anterior glands

and the last mentioned, the posterior glands, in the following discussion.

The posterior glands were called prostates in the original description of the

species by Smith (1895). Eisen (1896) followed the nomenclature of

Beddard (1895) and applied the term spermiducal to these same glands.

Michaelsen preferred to limit the term prostates to the glands which were

directly associated with sperm ducts and described the glands of S. eiseni

as prostate-like. The presence of the anterior glands was first noted by

Eisen (1896). He called them parietal glands. This name is in no way

suggestive of the function, neither are the posterior glands true prostates.

22 FLORENCE S. HAGUE

Consequently, since the anterior and posterior glands are similar in origin

and structure, and are related to the reproductive activities, which facts

will be shown subsequently, the writer has chosen to use the term, acces-

sory reproductive glands for both kinds, and to distinguish them from each

other by the adjectives, anterior and posterior. The term prostates will

be used for those glands which are connected with the sperm duct.

Each anterior gland (fig. 4) is a spherical or somewhat elongated mass

from which a duct passes through the body wall close beside seta a and

opens into the follicle of that seta. The glandular part (fig. 16) consists

of a tubular lumen surrounded by epithelial, muscular and glandular

layers; the duct lacks the glandular layer. The epithelial layer is a con-

tinuation of the body epidermis (fig. 18); the muscular, of the circular

muscular layer of the body wall. The proximal ends of the glandular

cells are attenuated and can be traced into the muscular layer. They

probably extend into the epithelial layer.

A posterior gland (fig. 3) consists of a tubular and somewhat convoluted

glandular part (fig. 14), and a duct (fig. 15) which opens to the exterior

close beside and usually through the follicle of seta b. The duct consists of

an epithelial layer which is continuous with the epidermis, and a muscular

layer. The peritoneum covering the duct is prominent. The glandular

part consists of epithelial cells with attenuated processes which extend

toward or to the lumen. Because of the regularity of cell walls and of

the position of the nuclei about the lumen there appears to be an epithe-

lial lining surrounded by a glandular layer.

The earliest stage in the development of an anterior gland was found

in specimen No. 68, which was collected in May and was without a trace

of a clitellum. In this gland (fig. 18) the epidermal invagination is distinct

and cells from the circular muscular layer can be seen extending part way

around it. Figure 19 is a section through the deepest part of the invagina-

tion, which is as deep as the thickness of the body wall. In later stages

mitotic figures are present in the epithehum. The epithehal nuclei are

very numerous and are arranged in two or three irregular series. The

muscular layer appears as a thin strand of cells which are not typical

muscle cells but which are connected with the circular muscular layer. At

this stage the glandular layer first appears. It consists of numerous nuclei

and a comparatively small amount of cytoplasm, but is without cell walls.

The nuclei are stained like those of the epithelium, are found close to the

muscular layer and occasionally are seen in sections directly over the

thin muscular strands. Because of these conditions and the attenuated

bases of the glandular cells in the fully developed gland, it seems that the

glandular cells are epithelial in origin. Nothing has been found which

would indicate their development from the body wall or from the coelomic

epithelium. In later stages the epithelial cells become definitely columnar

STUDIES ON SPARGANOPHILUS EISENI SMITH 23

and arranged in a single layer. The muscular and glandular layers gradu-

ally increase in thickness and cell walls appear in the latter. In the devel-

opment of a posterior gland, which is similar to that of an anterior one, the

continuity of the epithelial and glandular cells shows as soon as cell walls

appear.

Variations in size and position have been found in both kinds of glands.

Table IV gives the condition and location of these glands, in so far as they

were ascertained, in the various worms studied. Worms numbered from

one to 36 are in the collection of Professor Smith and the remainder have

been collected during the course of this study. The specimens from a

given locality are listed together in each of the two series. This table

shows that, so far as is known, the anterior and posterior glands in each

worm are in similar stages of development. Of the worms collected at

Homer Park in June and July, all have typical glands; in October, Decem-

ber and February, all have small glands. Furthermore, a distinct clitellum

is, without exception, accompanied by typical glands; an imperfectly

developed clitellum is most frequently, and an absence of clitellum, in

mature worms, is always accompanied by small glands. There is then a

decrease in the size of the glands between breeding seasons. This decrease

in size is not merely a shrinkage but an actual disintegration and loss of

certain layers. Muscular and glandular layers degenerate into a mass

(fig. 17, deg), in which cell outlines and nuclei are indistinct or wanting,

and at the surface of which are numerous bloodvessels. This mass dis-

appears leaving only the epithelial layer, which apparently does not degen-

erate, for mitosis occurs in its cells both before and after the loss of the

other layers. Mitotic figures have been found in the epithelium of worms

collected in December, February and May. The subsecjuent development

is similar to that already described. In the posterior glands there is a

similar process of degeneration and regeneration.

The original development of these glands is probably not closely re-

lated to the breeding season, for in Nos. 94 and 95, which were collected

in September, the glands were apparently developing. They were also

developing in No. 68 which was collected in May.

The posterior glands were found so regularly in three or four of somites

23-26 in the worms studied first, that, later, only somites 1-15 of most of

the worms were sectioned. Sometimes a part of somites 23-26 was sec-

tioned in order to ascertain the condition of the glands. Of all the worms

in which the location of the posterior glands has been investigated, only

specimens from Havana, Illinois have the glands in somites other than

23-26. Several of these worms have glands in three or four of somites 22-

27, and one or more of 15, 16 and 17.

Aside from the seasonal changes, there are in the anterior glands vari-

ations in size due chiefly to the thickness of the glandular layer; and vari-

24

FLORENCE S. HAGUE

Taule IV

Accessory Refroductive and Pharyngeal Glands

Some of the specimens were dissected: from one to 27 somites of the others were

sectioned. A blank means that the condition of the organ is unknown, t indicates a tj'pical

organ: 5, a small organ : 5^, a very small organ : 0, absence of the organ. The numbers which

follow these letters indicate the somites.

STUDIES ON SrARGANOPlIILUS EISENI SMITH

m

Table IV, continued

25

26

FLORENCE S. HAGUE

Table IV, continued

I

STUDIES ON SPARGANOPHILUS EISENI SMITH

27

Table IV, concluded

28 FLORENCE S. HAGUE

ations in shape, due to the configuration of the coelome and possibly to the

contraction of the muscles of the gland. In several worms there is a slight,

and in No. 151a very definite epidermal thickening surrounding the gland

pore. Typically, the anterior glands open into the follicles of setae a of the

somites in which the glands are located. These setae are not ornamented

or otherwise different from the usual type, but they are lacking in two

worms. No. 36 lacks setae a and has glands which are larger than usual

and which have a heavier muscular layer. No. 151 lacks setae a and b.

Since No. 36 is the only mature worm available from its locality, and No.

151 is the only one from its locality which was studied and which has

glands, it is not known whether their conditions are exceptional or usual

for their respective localities.

Variations in number and position of the anterior glands are numerous.

These glands occur most frequently in somites 3, 4 and 6. They are in

somite 7 of three worms; in somite 8 of two worms, and in somite 10 of one

worm. Most of the specimens which have the glands in 6 are from Havana,

Illinois (1920); most of those which have them in 4, from Homer Park,

Illinois and the earlier collections at Havana; most of those which have

them in 3, from Michigan and the earlier collections at Havana. There

may be a pair of glands symmetrically placed in the somite, or there may

be a gland on one side only. Each of about half of the worms from the

earlier collections at Havana has three or four glands in somites 3 and 4.

None of the recently collected worms have more than two glands, although

one of them (No. 142) has its two glands in two successive somites.

A summary of the condition of the anterior glands in the worms col-

lected in different localities shows that the glands are not always present.

Of a total of 46 worms from Homer Park, 39 have anterior glands in somite

4, and seven have no anterior glands; of 16 worms from Havana (1920)

eight have glands in 6; two, in 4; two, in 7; and four lack glands. Among

the Michigan specimens, the 22 from Sedge Pond have no anterior glands;

the same is true of five from Bessy Creek, two from the northwest shore

of the Lake, and six from Maple River. Of four worms from Hook Point,

two have glands in somite 3, and two lack glands; of eleven from Diogenes

Pond, seven have glands in 3, and four lack glands. Three worms from

Carp Creek lack glands and one has glands in 10. Of eleven worms from

the earlier collection at Havana, seven have glands in somites 3 and 4;

one, in 4; one, in 6; one, in 7; and one lacks glands. Of nine worms pre-

viously collected at Douglas Lake, one has an anterior gland in 4, and all

the others have glands in 3.

In all these groups mentioned, only certain ones from the Douglas

Lake region have a uniform condition in respect to the anterior glands.

These are the Sedge Pond, Bessey Creek, and Maple Ri\er groups, in all

of which such glands arc lacking. It is possible that a stud)- of more speci-

STUDIES ON SPARGANOPHILUS EISENI SMITH 29

mens from Bessey Creek and Maple River would result in finding some

worms with glands. Of the groups of four or more worms which were col-

lected in 1919 and 1920, and in some of which glands are present, the per-

centage having glands varies from 25 to 85. The significance of these

percentages lies in the fact that they show a wide variation. The position

of the glands is least variable in the worms from .Homer Park. All of these

have the glands in somite 4. The worms with the greatest variation in

the position of the glands are those from Havana. Those of the 1920 col-

lection have glands in somites 4, 6 and 7, with most of them in 6; those of

the earlier collections have glands in somites 3, 4, 6 and 7, with most of

them in 3 and 4.

The fact that glands, probably similar in function and differing only

slightly in structure, are found ventrally in different worms in several of

somites 3, 4, 6, 7, 8, 10, 15, 16, 17, 22, 23, 24, 25, 26 and 27 suggests that

these glands are remnants of a once continuous series of ventral glands.

Eisen (1896) wrote: "Of the same nature (as the spermiducal glands) I

consider the forward parietal glands in somite 3 of S. eiseni, and it seems

not unlikely that originally this genus possessed many more pairs of sper-

miducal glands, perhaps one in every somite." In order to account for the

two kinds of accessory glands, it would have to be assumed that there had

been two series of glands, or that one series had become differentiated into

two kinds of glands. The facts that all of the earlier (1913) collection

of worms from Douglas Lake, probably Diogenes Pond, have anterior

glands, but that over 2)Z 1/3% of those of the 1920 collection from Diogenes

Pond, which were studied, lack these glands; and that over half of the

earlier (1895) collection of worms from Havana had three or four glands,

while none of those from the recent collections have more than two glands,

suggest the possibility that these changes may be continuing even now.

Because of the variable condition of these glands, it scarcely seems that

their presence or absence can be regarded as a specific character.

The first mention in the literature of anything that seems comparable

to the accessory glands described above, is the description (Perrier 1874)

of 40 pairs of pyriform bodies located in the posterior somites of Ponto-

scolex (originally Urochaeta) corethrurns. Dissection and sections of a few

somites of a specimen of P. corethrurus, which is in the collection of Pro-

fessor Smith, have established the fact that each pyriform body is part of

a nephridium.

In the glossoscolecid genera, Andiorrhinus, Criodrilus, Kynotus, Micro-

chaeta, Rhinodrilus and Tritogenia, glands have been described which

are associated with the ventral setae. In Criodrilus these are conspicuous

glandular areas about each pair of ventral setae on the clitellum. Some of

the setae are modified as genital setae. In the genus Microchaeta, the

glands are located in some one or all of somites 9 to 35. There may be a

30 FLORENCE S. HAGUE

single gland or there may be several glands opening into a setal follicle.

Some of the setae are modified as genital setae, and sometimes external

papillae mark the location of the glands. The glands of M. papillata

(Benham 1892) and of a specimen of M. algoensis in the collection of Pro-

fessor Smith, are structurally like those of Sparganophilus eiseni, except

that those of Microchaeta algoensis have both circular and longitudinal

bands in the muscular layer.

Such structures are not unknown among Lumbricidae. The so-called

papillae, on which setae ah of 9, 10 and 11 of Helodrilus caliginosus trape-

zoides are located, are thickenings of the epidermis into which irregular

projections of the cavity of the setal follicle extend. There are glandular

structures accompanying the copulatory setae of Lumhricus terrestris

(Hering), and the ventral setae of 13-16 of Bimastns palustris (H. F.

Moore).

Glands are associated with ventral setae, which in some are modified

as genital setae, in the megascolecid genera, Acanthodrilus, Octochaetus,

Dichogaster, Diplocardia and Pheretima. Diplocardia (Eisen 1900) has

glandular areas consisting of cells which lie between the epidermal and

muscular layers and which have long ducts opening into the setal follicles.

In other genera the relation of the glands near the genital and spermathecal

orifices to the ventral setae is not known, but from brief accounts of their

structure it seems probable that they are similar to the accessory repro-

ductive glands described. The accessory glands described by Sweet are

not connected with the setae but with the sperm duct.

In one of the Moniligastridae, Syngenodrilus lamuensis (Smith and

Green), "prostates" have been described in somites 11-13. They are not

connected with the sperm ducts which open on the anterior margin of 13.

Each gland opens to the exterior slightly laterad of seta b, and has a tubu-

lar lumen surrounded by epithelial, glandular and muscular layers. Bed-

dard (1887) described a similar position for the muscular layer in the

prostates of Eudrilus eugeniae (originally sylvicola), but in all other glands

of the accessory type of which descriptions have been found, the muscular

layer is between the epithelial and glandular layers.

The glandular structures which have just been mentioned are so vari-

able and the data on their development and structure are so meager that

it is difficult to interpret their relations to each other. Lankester sug-

gested that the capsulogenous glands of Lumbricus were excessive develop-

ments of the setigerous glands. Beddard (1895) concluded that the

accessory, or copulatory glands as he called them, had developed as a

result of the invagination of a glandular area, and that the accessory and

prostate glands were serially homologous. It was suggested, also by

Beddard, that the glands may have developed differently in difTerent

families of earthworms. Stephenson and Haru concluded that the prostate

glands of Pheretima hawayana were mesodermal in origin, whereas it had

STUDIES ON SPARGANOPHILUS EISENI SMITH 31

previously been assumed that all prostates and accessory glands were

ectodermal. It has been shown, (p. 21) that the glands of Sparganophilus

eiseni are ectodermal in origin.

The accessory glands have been thought by different investigators to

produce slime, egg capsule (cocoon), albumen, a poisonous secretion, or an

irritating secretion. Rosa thought that the glands of Microchaeta benhami

had assumed a prostate function. Eisen (1896) held a similar view in

regard to the glands of Sparganophilus eiseni. Benham (1892) stated that

the copulating individuals of Lumbricus were held together by a sucking

action of the clitellum and not by a slime tube. He, accgrdingly, attributed

a sucking action to the accessory glands about the setae of Microchaeta

papillata. Buchanan suggested that the secretion of the accessory glands

which are near the spermiducal and oviducal pores of Notoscolex (origi-

nally Cryptodrilus) saccarius might aid in the passage of the body of the

worm through the cocoon without friction. Since these last mentioned

glands are not related to the setae, they may not belong to the same cate-

gory as those under discussion, and yet they seem to be similar to those

of Pheretima which Beddard (1895) considered capsulogenous.

No evidence for a definite conclusion in regard to the function of these

glands has been obtained from the present study. The writer did not see

any living specimens of Sparganophilus etseni in copulo, but did study

sections of a pair of worms tn copulo which had been collected by Professor

Smith at Havana, Illinois in 1895. As is generally the case in copulating

earthworms, the anterior ends are turned in opposite directions and are

closely approximated along their ventral surfaces. The ventral side of

somites 18-27 is concave and somites 1-9 of the opposite worm lie in this

concavity. These parts of the two worms are held closely together by a

slime tube which is constricted at both ends. There is such a slime tube

about one anterior end of each of two copulating pairs, but the other an-

terior ends have evidently pulled away from the opposed somites. It

is probable that the slime tube was continuous from somites 1-27 of both

worms.

The anterior end of the worm is pushed into the concavity of somites

18-27 so far that setae cd of somites 1-9 are just outside the ventro-lateral

edges of somites 18-27. Setae cd of somites 26 and 27 are on the convex

surface of those somites but close to their ventro-lateral edges. The

closest contact of the worms is in and immediately below the seta line, c,

of somites 1-9. In sagittal sections the epidermal cells of the two worms

are so intimately associated in places that the dividing line between them

is not readily recognizable. Medially there is an irregular slit-like space

between the worms.

The slime tube apparently is not formed as a single tube enclosing the

worms, but by the fusion of two tubes, one around each worm. A thicken-

ing in the slime tube, just laterad of the line of contact of the worms, is

32 FLORENCE S. HAGUE

interpreted as the line of fusion of the two tubes. While the slime, which

forms the tube enclosing the worms, is thin and deeply stained, that which

is between the worms is thicker and only faintly stained. Dorsally the

slime tube does not extend deep into all the narrow crevices of the inter-

segmental grooves, and does form a smooth, continuous outer surface,

which obliterates the outlines of segments. Ventrally the slime covering

extends into the various grooves, although not always closely, and forms

a small mass filling the anterior end of the slit-like space between the

worms. The cuticula is recognizable in different places, both dorsally

and ventrally, between the slime and the epidermis.

The accessory reproductive glands of somites 7, 25, 26 and 27 open

into the irregular slit-like space between the worms. The pores of the

accessory glands are not in contact with the body of the opposed worm,

therefore, these glands cannot be sucking organs as was suggested for

similar structures in Microchaeta papillata. The spermathecal pores open

very close to, but slightly laterad of the line of closest contact. The

spermiducal pore could not be found. The intersegmental grooves 19/20

and 8/9 are opposite each other. Since the accessory glands open into

the median space, from which the spermathecal pores are apparently

separated, it hardly seems that the secretion of the former can facilitate

the transfer of spermatozoa, if this transfer is accomplished as it is in

Lumbricus (Andrews).

Different investigators have thought that these glands helped in the for-

mation of the slime tube which is present on worms in copulo and at the

time of cocoon formation. Since the posterior glands are in somites at the

posterior end of the clitellum, and the anterior glands in somites opposite

the posterior end of the clitellum of worms in copulo, and since these glands

open on the ventral side where the clitellum is thin, it is possible that they

do produce a part of the slime tube, or the slime which plugs up the opening

at the anterior end of the worm. The glands of 15, 16 and 17, when present,

are between the clitella of the copulating worms and might help in the

completion of the slime tube. Or, the secretion of the accessory glands

might help to fasten the slime tubes together.

This study of the accessory reproductive glands shows that the pres-

ence and position, especially of the anterior glands, is variable; and that

the function of both anterior and posterior glands is related to the repro-

ductive activities but is not definitely known.

VI. Systematic Relations

Eisen (1896) first reported the presence of the anterior accessory

reproductive glands in SparganophUus eiseni. Since he did not find such

glands in other specimens of Sparganophilus from Central and North

America, he concluded that these glands distinguished 5. eiseni from all

other species. Because of the presence of such glands and of some minor

STUDIES ON SPARGANOPHILUS EISENI SMITH 33

diflferences, he separated S. eiseni from S. benhami and the less distinct

species, S. giiatemalensis and S. carneus. He also stated that he had in-

sufficient well-preserved material of the two latter species, and that both

might prove to be varieties of S. benhami.

The most important difference between S. benhami and S. eiseni, as

defined by Eisen, was the presence in the latter of the anterior accessory

reproductive (parietal) glands. In the present discussion of these glands

it has been concluded (p. 29) that they are too variable in presence and

absence to be properly regarded as a basis for separating species. These

species were also said to differ in size; in position of the tubercula puber-

tatis, and of the anterior nephridia; in the shape of the spermathecae; in

the lobulation of the sperm sacs and in the relative sizes of the pharyngeal

(septal) glands. Again facts have been presented to show that there is

quite a variation in each of these characters among the specimens of S.

eiseni studied, and that there are conditions among some of these specimens

which are similar to the conditions described in S. benhami. A specimen,

identified by Eisen as S. benhami, is in the collection of Professor Smith.

It does not have the clitellum limited ventrally to somites 17-26, as de-

scribed for the species, but on 15-27, with a thicker portion on 22-27.

It also has the spermiducal pores on somite 19, not on 20. It has no dorsal

pores. Sections reveal nothing which is different from conditions found

in S. eiseni. Therefore it seems that S. benhami should be united with

S. eiseni.

Since the presence of anterior glands is insufficient to distinguish

species, the only significant difference between S. guatemalensis and ^S.

eiseni is the fact that the clitellum in the former is on somites 16-26. In a

specimen of S. eiseni collected in October, the clitellum appears to begin

on somite 16, probably because the clitellum was degenerating. Since the

material, on which the description of S. guatemalensis was based, was not

in good condition; since the difference is small, and especially since, in other

points, it agrees with the conditions found in S. eiseni, there seems to be

insufficient basis for making this a separate species. The differences

between S. carneus and S. eiseni are also such as may be accounted for in

the variations of the latter. Eisen suggested that S. carneus might be a

northern form of S. benhami, and mentioned that the former resembled

S. eiseni in the shape of the spermathecae.

In the original description of the genus Sparganophilus, Benham placed

it in the family Rhinodrilidae, which he had defined (1890), but which has

since been made a part of the family Glossoscolecidae. Michaelsen (1917)

united the Glossoscolecidae and the Lumbricidae into the Lumbricidae

s. I., and subdivided the former Glossoscolecidae into five subfamilies:

Glossoscolecinae, . Sparganophilinae, Microchaetinae, Criodrilinae and

Hormogastrinae. These were given equal rank with the Lumbricinae,

formerly Lumbricidae, in the new family Lumbricidae s. I. This was done

34 FLORENCE S. HAGUE

because additional studies had made a separation of the Glossoscolecidae

and Lumbricidae, as two families, seem to him impracticable. According

to this classification, Sparganophilus in the only genus of the subfamily

Sparganophilinae. Whereas, it was formerly considered the ancestral

form of the Glossoscolecidae, Michaelsen now considers it a degenerate

descendant of the Glossoscolecinae. In a more recent paper (1921),

Michaelsen has created a Familienreihe Lumbricina, in which he has placed

the families Glossoscolecidae, Sparganophilidae, Microchaetidae, Hormo-

gastridae, Criodrilidae and Lumbricidae.

VII. Summary

1. The embryology, as far as it was followed, presents no marked

differences from that of other earthworms.

2. Aside from the somewhat greater histological differentiation, the

nephridium of Sparganophilus eiseni is similar to that part of the nephrid-

ium of Lumbricus, which extends from the nephrostome to the muscular

duct. There is no muscular duct in the nephridium of Sparganophilus

eiseni.

3. The nephridia begin to develop in somite 3 and the somites poste-

rior thereto in embryos of 5. eiseni. Disintegration soon sets in at the

anterior end, and causes the complete loss of the nephridia of somites 3-11

and the loss or degeneration of the nephridia of 12 and 14, and sometimes

of 13.

4. The genital ducts and funnels, although closely associated with the

nephridia, develop independently of them.

5. The accessory reproductive glands, of the various specimens ex-

amined, are found in three or more of 15 different somites, and probably

are remnants of a once continuous series of glands. The anterior accessory

reproductive glands are too variable to be of value in distinguishing spe-

cies.

6. Sparganophilus eiseni is a variable species in several respects.

Since each of the combinations of structural characters which are included

in the descriptions of the various species: S. hcnhami, S. guatemalcnsis

and S. carneus are found represented among various individuals of S.

eiseni, it seems necessary to unite them into the one species, .S". eiseni.

VIII. Literature Cited

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STUDIES ON SPARGANOPHILUS EISENI SMITH 35

Beddard, F. E.

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Benham, W. B.

1886 Studies on Earthworms. Quart. Journ. INIicros. Sci.,' 26:213-301.

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1912 The Nephridia of Perieodrilus ricardi and of P. montanus. Trans. N. Zealand

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Bergh, R. S.

1886- Untersuchung liber den Bau und die Entwickelung der Geschlectsorgane der

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1899 Nochmals iiber die Entwicklung der Segmentalorgane. Zeit. wiss. Zool., 66:435-

449.

Bourne, A. G.

1894a On Certain Points in the Development and Anatomy of some Earthworms.

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1894b On JMoniligaster grandis, A. G. B. from the Nilgiris, S. India: together with

Descriptions of other Species of the Genus Moniligaster. Quart. Journ. Micros.

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Buchanan, G.

1911 Note on the Accessory Glands of Cryptodrilus saccarius (Fletcher). Proc. Roy.

.Soc. Victoria, 22:221-223.

36 FLORENCE S. HAGUE

Cameron, Gladys M.

1912 The Minute Structure of the Nephridium of the Earthworm Maoridrilus rosae

Bcddc-rd. Trans. N. Zealand Inst., 45:172-190.

Cole, F. J.

1893 Notes on the Clitellum of the Earthworm. Zool. Anz., 16: 440-446.

Collin, Ant.

1888 Criodrihis lacuum Hoffrn. Zeit. wiss. Zool., 46:471-497.

ElSEN, G.

1895 Pacific Coast Oligochaeta. I. Mem. California Acad. Sci., 2:63-122.

1896 Pacific Coast Oligochaeta. II. Mem. California Acad. Sci., 2:123-198.

1900 Researches in American Oligochaeta, with Especial Reference to those of the

Pacific Coast and Adjacent Islands. Proc. California Acad. Sci., (3) 2:85-274.

Foot, K. and Strobell, E. C.

1902 Further Notes on the Cocoons of AUolobophora foetida. Biol. Bull., 3:206-213.

Gatentby, J. B.

1916 The Development of the Sperm Duct, Oviduct and Spermatheca in Tubifex

rivulorum. Quart. Journ. Micros. Sci., 61:317-336.

Gegenbauer, C.

1853 Ueber die sogenannten Respirationsorgane des Regenwurms. Zeit. wiss. Zool.,

4:221-232.

GOEHLICH, G.

1890 Uber die Genital- und Segmental-Organe von Lumbricus terrestris. Zool.

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Goodrich, E. S.

1895 On the Coelom, Genital Ducts and Nephridia. Quart. Journ. Micros. Sci.,

37:477-510.

Heimberger, H. V.

1915 Notes on Indiana Earthworms. Proc. Indiana .\cad. Sci., 1914:281-285.

Hering, E.

1857 Zur Anatomic und Physiologie der Generationsorgane des Regenwurms. Zeit.

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Hesse, R.

1894 tjber die Septaldriisen der Oligochaetcn. Zool. Anz., 17:317-321.

HORST, R.

1888 Descriptions of Earthworms. Notes Leyden Mus., 10:123-128.

1895 Descriptions of Earthworms. Notes Leyden Mus., 17:21-27.

Lankester, E. R.

1864 The Anatomy of the Earthworm. Quart. Journ. Micros. Sci., 4:258-268; 5:7-18,

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Lehmann, O.

1887 Beitrage zur Frage von der Homolgie der Scgmentalorgane und Ausfuhrgange der

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1910 Die Excretionsorgane der wirbellosen Ticrc. Ergebn. und Fortsch. Zool., 2:275-

366.

MiCHAELSEN, W.

1891a Tcrricolcn der Berliner Zoologischen Sammlung. I. Afrika. Arch. Naturg., 57,

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1891b Bcschreibung der von Ilcrrn Dr. Fr. Stuhlmann auf Sansibar und dem gogcniiber-

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9:1-72.

STUDIES ON SPARGANOPHILUS EISENI SMITH 37

1892 Terricolen der Berliner Zoologischen Sammlung. II. Arch. Naturg., 58, 1 :209-261.

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1921 Zur Stammesgeschichte und Sj^stematik der Oligochaeten, insbesondere der

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1911 Der Wimpertrichter der Lumbriciden. Zeit. wiss. Zool., 98:135-178.

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1889 Etudes sur le developpement des Annelides. Ann. sci. nat. [Zool.], (7) 7:107-442.

ScHNEroER, Karl C.

1902 Lehrbuch der vergleichenden Histologic der Tiere. XIV+988 pp. Jena.

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1895 A preliminary Account of two New Oligochaeta from Illinois. Bull. Illinois

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1919 Descriptions of New African Earthworms, including a New Genus of Moniliga-

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1910 Organogenetische Untersuchungen ueber Criodrilus lacuum Hoffmstr. Arb.

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38

FLORENCE S. HAGUE

1917 On the So-called Pharyngeal Gland-cells of Earthworms. Quart. Journ. Micros.

Sci., 62:253-286.

Stephenson, J. and Haru, Ram.

1919 The prostate glands of the Earthworms of the Family Megascolecidae. Trans.

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Sweet, Georgina.

1900 On the Structure of the Spermiducal Glands and Associated parts in Australian

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Vejdovsky, F.

1884 System und Morphologie der Oligochaeten. 166 pp. Prag.

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Wilson, E. B.

1889 The Embrj-olog}- of the Earthworm. Journ. Morph., 3:387-462.

EXPLANATION OF PLATES

All drawings except figures 1 and 2 were made with the aid of a camera lucida.

''/

dor

X45.

Plate I

Fig. 1. Outline of tlie bifurcation in a young worm. Dorsal view.

Fig. 2. Outline of a nephridium and its canal.

Fig. 3. A somite with a posterior accessory reproductive gland. Longitudinal section.

Fig. 4. Somite 4 with an anterior accessory reproductive gland. Cross section. X 45.

epn

[0^ cm

wi '^W-^. ©pn

Wl^ — c

ne

Plate II

l-'ig. 5. Section through the wide tube at i (fig. 2). X 46*).

Fig. 6. Part of a section through a ne[)hridium. X 469.

Fig. 7. Section through the wide tube at its entrance into,the body wall. X 46A

Fig. 8. Section through the ncphridioporc. X 409.

10

13

Plate III

Fig. 9. Section through the junction of middle and wide tubes. X 469.

Fig. 10. Section through a developing nephridium. X 1173.

Fig. 11. Section through a developing nephridium of a 0.5 mm. embryo: an earlier

stage than figure 10. X 1173.

Fig. 12. Section through a ty-pical young nephridium. X 1173.

Fig. 13. Part of a section through a disintegrating nephridium. X 1173.

IS

Im— ^.

cm

Plate IV

Fig. 14. Posterior accessory reproductive gland ; part of a section througli the glandular

portion. X 300.

Fig. 15. Posterior accessor}- reproductive gland; section through the transition of duct

into glandular portion. X 300.

Fig. 16. Anterior accessory reproductive gland; part of a section tlirough the glandular

portion. X 300.

Fig. 17. Part of a section through a degenerate anterior accessor}- reproductive gland.

X514. - ^ ■ y A

Figs. 18 and 19. Sections through a developing anterior accessory reproductive glana.

Fig. 18. The invagination. X 514.

Fig. 19. The inner part. X 514.

A SYSTEMATIC PRESENTATION OF NEW GENERA

OF FUNGI

By

O. A. Plunkett, p. a. Young, and Ruth W. Ryan

University of Illinois

The new families and genera of fungi described since volume 22 of

Saccardo's "Sylloge Fungorum" was compiled are here assembled from

all available literature and presented in a concise, classified form with the

reference accompanying each new name. This paper is necessarily incom-

plete because some publications and parts of others were unavailable.

Only small parts of Broteria, The Botanical Magazine of Tokyo, and

Osterreichische Botanische Zeitschrift were found.

As far as is known, there has been no previous compilation of the new

genera of fungi described since 1910. Mycologists have been compelled to

search through an extensive, scattered literature for the special types in

which they were interested. This paper will be of value to mycologists in

that it contains, with necessary references, most of the new genera of

fungi published since the last volume of the "Sylloge Fungorum" was

compiled.

The authors wish to thank Dr. F. L. Stevens for numerous helpful

suggestions. To him is due the credit for planning the work of assembling

all the new families, genera, and species of fungi described since 1910.

They also wish to acknowledge the assistance of Messrs H. L. Dixon,

J. M. Mendoza, P. J. Byrd, H. C. Abbott, and Miss Ruth Dowell who

aided in the search through the literature.

Approximately 7,000 new species of fungi were listed on cards and

catalogued in taxonomic order. The cards bear the citation, classification,

the name of the genus, species, and generally the host of the fungus. Of

these about 800 belong in the Sphaerioidaceae, 700 in the Agaricaceae,

300 in the Pucciniaceae, 200 in the Dematiaceae, 200 in the Microthy-

riaceae, 200 in the Pleosporaceae, 150 in the Mycosphaerellaceae, and

100 in each of the following families: Dothideaceae, Hypocreaceae, Melan-

coniaceae, Moniliaceae, Polyporaceae, Sphaericaeae, Thelephoraceae,

Tuberculariaceae, and Valsaceae. This list is too voluminous to print

at the present time.

Explanation of Tables

The name of the new genus is given in the first column. The name in

the second column is that of a genus nearly related or similar to the new

43

44 O. A. PLUNKETT, P. A. YOUNG, AND RUTH W. RYAN

genus named in column 1 or sometimes some other significant name.

A number has been given to each publication. This number is given in

the 3rd column and refers to the bibliography. The number in column 4

refers to the volume, that in column 5 to the page, and the one in column 6

to the date of publication.

At the end of the list is a group of new genera the positions of which

were not definitely stated by the authors and the afiinities of which were

not sufficiently clear to warrant us in assuming their relationships.

The classification system used in this paper is mainly that of Engler

and Prantl as given in "Die Naturlichen Pflanzenfamilien" except in the

Dothideales and Hemisphaeriales in which the keys of Theissen and Sydow

are followed as closely as possible.

For the most part, the genera are classified where the authors placed

them even though the positions of many were subsequently changed.

See the "Synoptische Tafeln" of F. Theissen and H. Sydow in Annales

Mycologici 15:389-491. 1917.

M\^OMYCETES

Plasmodiophorales

Plasmodiophoraceae

Anisomyxa 23

Clathrosorus 9

IMolliardia 7

Ostenfeldiella 9

Sorodiscus 11

Sorolpidium 23

Trematophlyctis 54

PHYCOMYCETES

Chytridiales

Synchytriaceae

Mitochytridium 54 27 202 191 1

Monochytrium Synchytrium 45 10 3, 50 1910

Saprolegniales

' Saprolegniaceae

Isoachlya . .Achlya 3

Jaraia 23

Pythiomorpha 68

Rheosporangiuni 82

Leptomilaccae

AUomyces Blastocksia 9 25 1023 1911

A PRESENTATION OF NEW GENERA OF FUNGI 45

Peronosporales

Peronosporaceae

Bremiella Brcmia 41 6 195 1914

Nozemia 52 13 566

Stigeosporium Phytophthora 9 30 357 1916

MUCORALES

Mucoraceae

Blaskestca Choanophora 18

Dissophora Mortierella 18

Haplosporangium Mortierella 18

PHYCOMYCETES STERILIA

Zoophagus 46 61 368 1911

ASCOMYCETES

Protomycetales

Hemiascaceae

Dipodascus 85 5 1 1921

Taphridium 85 5 1 1921

Saccharomycetales

Endogonaceae

Protascus 42 3 155 1913

Saccharomycetaceae

Aleurodomyces 13

Blastocystis Dermatocystis 28

Coccidiascus 28

Cycadomyces 73

Guillermondia 65

Medusomyces Mycoderma 16

Nectaromyces 22

Psillidomyces 13

Helvellales

Helvellaceae

Geomorum 89 23 1921

Pezizales

Pezizaceae

Aleurina Aleuria 41

Aparyphysaria 89

Manilaea 7

Mollisina 16

Pezizellaster Pezizella 7

46 O. A. PLUNKETT, P. A. YOUNG, AND RUTH W. RYAN

Helotiaceae

Belonioscyphella Belonium 53 127 590 1918

Calycellina Helotium 53 127 601 1918

Helotiopsis Mollisiella 53 119 623 1910

Lachnobelonium Belonium 16 37 109 1919

Lambertella Belonioscypha 53 127 375 1918

Leptobelonium Belonidium 16 37 108 1919

Microscypha Dasyscypha 7 17 38 1919

Neobulgaria Ombrophila 7 19 44 1921

Psilachnum 16 37 109 1919

Stereolachnea Lachnea 7 15 353 1917

Tanglella 53 127 606 1919

Torrendiella Dasyscypha 54 27 133 1911

Patelliariaceae

Pleoscutula Scutula 54 29 434 1913

Rodwaya Woodiella 86 13 425 1917

Siscocera Nesolechia 59 6 48 1917

Cenangiaceae

(Including Bulgariaceae)

Asterocalyx 53 121 402 1912

Bulgariastrum 47 8 497 1913

Caloriopsis 7 15 254 1917

Caloriella Cenangina .53 127 345 1918

Discomycella Ascosorus. . 53 121 401 1912

Encoeliella 53 119 619 1910

Stegopeziza Dermataceae 53 126 308 1917

Phacidiales

Stictidiaceae

Eupropolella 7

Hysteropezizella Stegia? 53

Hystciostegiella Sarcotrochila 53

Phacidiella Char, emend 53

Propoliopsis 36

Sarcotrochila Trochila 53

Tryblidiaceae

Odontoschizon Helerosphaeria . 7 12 568 1914

Phacidiaceae

Leptophacidium 53

Myxophacidiella 53

Myxophacidium '. 53

Phacidiella 67

Hysteriales

Hypodermataceae

Bifusella Hypoderma 7

Haplophyse 7

Hypodermellina Hypoderma 7

A PRESENTATION OF NEW GENERA OF FUNGI 47

Hysteriaceae

Hysterostomina Hysterostomella 7

Parmulina Parmularia 7

Periaster Erikssonia 7

Polhysterium Hysterographium 8

TUBERALES

Tuberaceae

Hydnotryopsis Geopora 62 6 336 1916

Agyriaceae

Ramosidla 7 15 254 1915

ASPERGILLALES

Asptrgillaceae

Acaulium 64

Aspergillopsis 64

Crollium 64

Rhopalocystis 35

Periosporales

Erysiphaceae

Chilemyces

]•';

Leucoconis 7

Schistodes nov. nom 7

Typhulochaeta 19

Perisporaceae

Acanthostoma 15

Chaetostigme Dimeriella 7

Chaetostigmella Phaeodimeriella 7

Cleistosphaeria 7

Diblastospermella Dimeriorum 77

Dichothrix 15

Dimeriopsis Dimerium 60

Eosphaeria 7

Euantennaria 77

Eudimeriolum 8

Guttularia Orbicula 42

Haraea 7

Jaffuela 77

Meliolina 7

Nematothecium 36

ParodielJa Char, emend 7

Parodiopsis Parodiella 55

Perisporiopsis Perisporium 60

Phaeodimeriella 15

Phaeostigme Dimerium 7

Phycopsis Seuratia 27

Pilene 7

48 O. A. PLUNKETT, P. A. YOUNG, AND RUTH \V. RYAN

Plenophysa 7 17 142 1919

Pseudoparodia Dimerosporina 7 15 138 1917

Rizalia ; . 7 12 546 1914

Rhizogene Lasiobotrys 7 18 181 1920

Rhizosphaerella Perisporium 32 59 254 1917

Setella 7 14 359 1916

Stigme Dimerina 7 15 199 1917

Stomatogene 7 14 404 1916

Teratonema 7 15 180 1917

Trichospermella 8 23 38 1912

Winteromyces Parodiclla 8 23 37 1912

Englerulaceae 81 66 296 1916

Diathrypton 47 21 137 1922

Euthrypton 70 66 296 1916

Linotexis Parcnglerula 7 15 198 1917

Oothecium 77 23 519 1918

Ophiolexis 70 66 296 1916

PhaeoschifiFnerula 20 12 21 1914

Rhizotexis 7 15 141 1917

Syntexis 70 66 296 1916

Theissenula Schiffnerula 7 12 198 1914

Thrauste 70 66 296 1916

Capnodiaceae

Adelopus nov. nom 7 15 482 1917

Aithaloderma 7 11 257 1913

Antenella 7 15 473 1917

Balladynopsis Balladyna 7 15 435 1917

Balladynella 7 15 478 1917

Calyptra 7 15 478 1917

Ceratochaete Sitella 7 15 179 1917

Chaetothyrina 7 11 495 1913

Chrysomyces 7 15 475 1917

Crytopus 7 12 72 1914

Microtyle .77 23 458 1918

Neohoehnelia 7 15 476 1917

Phragmicapnis 7 15 480 1917

Schizocapnodiiim Capnodium 49 6 91 1921

Parodiellaceae

Acantharia 7 16 15 1918

Epiphyma 70 306 1916

HjTJophlegma 7 15 135 1917

Parodiellinaccae N. Earn . . . . 4 16 21 1918

HEMISPHAERIALKS

I Ifmisphaeriaccac

Chaetoplaca 7 15 232 1917

Dictothyriella 20 12 92 1914

Dictyothyrium Dictyopeltis 46 62 277 1912

Epipeltis 1 7 3,26 1913

Eremotheca Eremothecella 7 15 235 1917

416 1911

15 146 1917

A PRESENTATION OF NEW GENERA OF FUNGI 49

Eremothecella Phragmothyriella 7

Haplopeltis 20

Hormopeltis o

Myiocoprella ^

Plochmopeltis 20

Polyclypeolum Microthyriella 7

Stephanotheca 47

Stomiopeltis 20

Stomiopeltella 20

Trichopeltaceae N. Fam 26

Pycnocarpon j

Pycnoderma ^

Trichopeltina 2 c

Trichopeltula 26

Trichosphaeriaceae

Melanopsomella 7 J7^ J21 1919

Microthyriaceae

Actinocymbe 15 ^ .20

Actinomyxa 7

f'^'^^p^^^'^ •■■■■••'•^^■'^^■■^■^■::':::::::::::: 7 n 315 1913

^TZ^- 7 11 499 1913

Asterolibertia Dimerosporium 4 16 166 1918

Asteromyxa 7 j5 ^^^ ^^^^

Asterostemula Asterinella. ... 7 14 270 1916

^T'^^'^tu 20 10 101 1912

Auographiella 7 ^5 3^^ ^^^^

^"""^'^"•; •. 4 16 123 1918

Calothyriella Calothyrium 7 15 371 1917

Calothyrium / 7 10 160 1912

r Au 120 82 1914

?"^^"^: 7 14 90 1916

Caenothyrium 7 j^ ^j^ ^^j^

Lamposa 77

Chaetothyrium 77

^^^^°^i",; ■•■:■■:;;::;■;::::::::::::;::::: 4 i6 ^ 1918

^r^^'ft 4 16 129 1918

^^yP'^^'^^^ 53 119 403 1910

Clypeolina /26 34 234 1912

^ . . 1 7 15 419 1917

Locconiopsis .

Dictyothyrium

Echiclnodes Lembosia. '. ^' .'.'.''.' .' 7 15 422 1917

Echidnodella Morenoella 7 15 422 1917

l-f'f^'^^ 53 119 454 1910

^"°Pf^ 53 119 420 1910

!!t^"!^"^ 7 14 430 1916

2"^?^"T Halbania 4 16 163 1918

?^"^^"^^ Microthyrium 27 164 890 1917

^'^""^ ■ Meliola 7 15 194 1917

25 90 1921

23 522 1918

16 113 1918

46 62 277 1912

50 O. A. PLUNKETT, P. A. YOUNG, AND RUTH \V. RYAN

Lembosia Char, emend 7 11 427 1913

Lembosina 7 11 437 1913

Lembosiopsis 7 11 435 1913

Kriegeriella 7 16 39 1918

Maublancia Asterina 4 16 159 1918

Maurodothella 4 16 124 1918

Melanochlamys 39 438 1913

Meliolaster 61 8 123 1919

Micropeltella 7 11 405 1913

Morenoina 7 11 434 1913

Mycolangloisia Lembosia 4 16 157 1918

Niesslella 16 36 468 1918

Parengkrula 53 119 465 1910

Patouillardina 27 164 890 1917

Peltella Myiocopron 7 15 237 1917

Phaeopeltis 83 7 1 1919

Prillieuxina Asternella 4 16 161 1918

Protothyrium 27 164 575 1917

Pycnoderma 7 12 563 1914

Pycnopeltis 7 14 365 1916

Questeria Dimerosporium 4 16 186 1918

Seynesiella Myiocopron 4 16 196 1918

Sirothyriella 53 119 451 1910

Stegothyrium 53 127 382 1918

Symphaeophj-ma Microphaeophyma 8 23 97 1912

Symphaster 7 13 217 1915

Thallochaete 7 11 501 1913

Thyrosoma 7 19 307 1921

Trichasterina Asterina 4 16 172 1918

Wardina Asterina 4 16 165 1918

Yatesula Stephanotheca 7 15 237 1917

Microthyriopsidaceae N. Fam. 4 16 99 1918

Leprieurina 4 16 210 1918

Manginula : 4 16 99 1918

Trichothyriaceae 15 32 3 1914

Trichothyriella 15 32 4 1914

Trichothyriopsis 15 32 4 1914

Polystomellaceae 7 13 158 1915

Armalella Polyrhizon 7 13 235 1915

Asterodothis 7 10 179 1912

Chaetaspis Parmulina 7 15 279 1917

Cyclotheca 7 12 7 1914

DothithjTcUa 7 16 171 1918

Ellisiodothis , 7 12 73 1914

Hysterostomina Ilystcrostomella 7 13 228 1915

Inocyclus Polycyclus 7 13 211 1915

Marchalia Char. Emend 7 13 251 1915

Mclanoplaca Marchalia 7 15 29S 1917

Monorhiza uleopcltis 7 13 318 1915

Monorhizina Monorhiza 7 13 320 1915

Pleiostomella Uleopcltis 7 15 221 1917

A PRESENTATION OF NEW GENERA OF FUNGI 51

Poiycjclina Polycyclus 7

Rhipidocarpon { -

Scoleoncma IJothidastromclla 7

Synpeltis 7

Stigmataceae N. Fam 7

Aphysa 7

Isomunkia Coccinopeltis 7

Vizdla 20

Hypocreales

Hj'pocreaceae

Balansiopsis Balansia 53

Borinquenia 60

Bronectria 77

Chromocrca 41

Chromocreophis 41

Cylindrocarpon Nectria 69

Dextria Caloncctria 60

Dialhypocrea 77

Epinectria 7

Epispora 54

Hyalocrea 7

Hyalosphaera 60

Hypocreophis 77

Leptocrea 7

Linearis troma 53

Mastigocladium 27

Nectriopsis Nectria 7

Neonectria Nectria 7

Orcadia 59

Patellonectria 77

Phy llocrea 7

Podonectria Ophinectria 59

Sterocrea 7

Trailia 59

Uropolystigma Polystigma 54

Dothideales

Plectodiscellaceae N. Fam. . .42

Plectodiscella 42

Myriangiaeceae 7

Ascostratum 7

Butleria 7

Symphaeophyma '. 8

Myxomyriangiaceae

AT • • / 7

Myxomyriangium < „

Saccardiaceae

Byssogene 47

Calopeziza .47

52 O. A. PLUNKETT, P. A. YOUNG, AND RUTH W. RYAN

Dothideaceae Char, emend.. . 7

Achorella Sj'stremma 7

Actinodothis 47

Amerodothis 7

Angatia 7

Auerswaldiella 7

Aulacostroma 47

Bagnisiopsis ; 7

Benguetia 7

Botry ostroma 53

Castagnella 54

Catabotrys Amerodothis 7

Catacauma Sphaerodothis 7

Clypeostroma 7

Coccochora Char, emend 53

Coccochorella 53

Coccodothis Phyllachora 7

CoccodotheUa Coccodothis 7

Coccostroma Yoshinagella 7

Cyclodothis 7

Cyclotheca 7

Dictyochora Curreya 7

Dictyodothis Phragmodothis 7

Diplochora 7

'53

Dothidastromella , „

Dothideopsella 53

Dothidina 7

Pvllisiodothis Asterula 7

Ehnerococcum Coccidella 7

Englerodothis Leveillella 7

Halstedia 18

Haplodothis , 53

Haplotheciella 53

Heterodothis 47

Hysterostoma 7

Ixptodothis 7

LeveilHna 7

Leveillella 7

Melanopsamnopsis Dothidella 76

Metachora 7

IVIetameris Achorella 7

Microdothella 47

IMicrocylella 7

Microphrodothis Ophiodothis 77

Ophiodothiella Ophiodothis 53

Palawania 47

Parmulina Parmularia 7

Perischizon Dothidea 7

Phaeodothiopsis Phaeodothis 7

Phragmodothella Achorella 7

A PRESENTATION OF NEW GENERA OF FUNGI 53

Phragmodothis 7

Phragmosperma 7

Phyllachorella 7

Placostroma 7

Polyrhizon 7

Psalidosperma 7

Pseudophyllachora 77

Pseudosphaerella Montagnella 53

Puiggarina 77

Pyrenobotrys 7

Rhabdostroma Scirrhiella 7

S6

Schizochora , „

7

Septomazzantia Mazzantia 7

Stalagmites 7

Stigma todothis 47

Stigmochora 7

Trabutiella 7

Trichochora Discodothis 7

Trichodothis 7

Uleodothis Auerswaldia 7

Uleodothella 7

Yoshinagella 53

Yoshinagamyces Yoshingaia 19

Phj'llachoraceae

Anisochora Catacauma 7

Apiospora Char.-emend 7

Camarotella Trabutia 7

Catacaumella Catacauma 7

7

Derma todothis , ^

Dictyochorella Dictyochora 7

Endothiella Phyllachora 7

Municidothis Trabutia 7

Omphalospora Munkidothis 7

Phaeochorella Catacauma 7

Phaeotrabutiella Trabutiella 7

Phragmocarpella Phyllachora 7

Phragmocauma Catacauma 7

Podoplaconema Omphalospora 7

■ 7

Rhemiodothis Trabutia i -

Rhopographina Rhopographus 7

Scirrhodothis Scirrhia 7

Scirrophragma Scirrhia 7

7

Scolecodothis .

7

Trabutia • Char, emend 7

Thyriopsis 7

54 O. A. PLUNKETT, P. A. YOUNG, AND RUTH W. RYAN

Montagnellaceae

Crotone 7

Epibotrj^s 7

Haplothecium 7

Hyalocurreya 7

Monopus 7

Montagnellina Montagnella 53

Ophiocarpella 7

Rosenscheldiella 7

Scirrhiachora Diplochorella 7

Syncarpella Montagnella 7

Pseudosphaeriaceae

Epiphyma 70

Lasiostemma 7

Monascostroma 7

Pseudoplea 7

Pyreniella 70

Scleropolella 7

Sphaeriales

Chaetomiaceae

Peristomium 27 55 178 1912

Sordariaceae

Fimetaria 43 3 65 1910

Coronophoraceae

Eucanthe Coronophora 7

Heteropera 7

Sphaeriaceae

Apiosporina 53

Asterosphaeriella Didymosphaeria 7

Bakeromyces 7

Bolosphaera 7

Boydia 59

Coccidophthora 7

Dasysphaeria 8

Dimerinopsis 7

Entopeltis 53

Gibsonia Spermatoria 9

Griposphaeria 7

Haplovalsaria Valsaria 53

Herpotrichiella . . Acanthostigma 7

Henningsomyces Char, emend 53

Linobolus 7

Meringosphaeria 44

Nematostigma 7

Nematostoma 7

Neokcissleria 7

NeotroUeria 74

A PRESENTATION OF NEW GENERA OF FUNGI 55

Plactogene 7 14 423 1916

Plagiostromella 53 126 372 1917

Porostigme Dimerinopsis 7 15 202 1917

Pseudopleospora 7 17 84 1919

Wageria Acanthostigma 41 11 8 1919

Xenothecium 53 128 589 1919

PhysosporellaceaeN. Fam...53 128 557 1919

Lejosphaerella Physosporella 53 128 577 1919

Ceratostomataceae

Chaetoceralostoma 14 2 144 1912

Cyanospora 41 2 209 1910

Cucurbitariaceae

Cucurbidortris Cucurbitaria 7 19 201 1921

Cucurbitariella 7 14 440 1916

Keissleriella Otthiella 53 128 582 1919

'53 119 417 1910

Montagnina ,^3 ^,1 350 1912

Rostronischkia 41 11 166 1919

Amphisphaeriaceae

StarbackieUa 7 17 37 1919

Titanella 7 17 36 1919

Mycosphaerellaceae

Discosphaerina Guignardia 53 126 353 1917

Lulworthia 59 5 259 1916

Pleosporaceae

Acantharia Hypolegma 7 16 15 1918

Acanthotheciella Ophiochaeta 53 120 450 1911

Crisserosphaeria Ophiobolus 8 23 72 1912

Didymellina Did>Tnella 7 16 66 1918

Epipolaeum Hypolyma 7 16 7 1918

Phanerococcus Hypoplegma 7 16 9 1918

Physalosporina Physalospora 7 9 288 1911

Plectosphaeria Physalospora 7 14 413 1916

Scleropliella Leptosphaeria 7 16 158 1918

Massariaceae

Leptomassaria Phorcys 7 12 474 1914

Myelosperma Pseudomassaria 7 13 38 1915

Trematosphaeria Massaria 53 123 99 1914

Gnomoniaceae

Desmotascus Phomatospora 18 68 476 1919

Stegasphaeriaceae N. Fam. . . 7 14 364 1916

Stegasphaeria 7 14 362 1916

Clypeosphaeriaceae

Amerostegi 70 66 297 1915

Euacanthe Clj^osphaeria 7 15 272 1917

Linocarpon 7 15 210 1917

Schizostege 7 14 415 1916

56 O. A. PLUNKETT, P. A. YOUNG, AND RUTH W. RYAN

Stigastroma 7 14 81 1916

Teratosphaeria 7 10 39 1912

Trabutiella Trabutia 18 70 401 1920

Valsaceae

Allanthoporthe Diaporthe 32 62 289 1920

Clypeoporthe Diaporthe 53 128 584 1919

Discoidaporthe AUantoporthe 32 62 293 1920

Macrodiaporthe Diaporthe 7 17 94 1919

Phaeodiaporthe Diaporthe 7 17 99 1919

Valseutypella Valsa 7 18 72 1920

JMelanconidiaceae

Amphicytostroma Crytosporella 7 19 63 1921

Cryptoceuthospora Crytosporella . 7 19 56 1921

Diatrypaceae

Apioporthe Diatrype 53 126 381 1917

Ectosphaeria Diatrype 77 25 48 1921

Phaeotrj-pe Diatrype 41 12 201 1920

Melogranimataceae

Anisomyces 7 12 270 1914

Causalis 7 16 184 1918

Pseudothis 7 12 274 1914

Phaeobotryon Botryosphaeria 7 13 664 1915

Xj'lariaceae

Theissenia 54 30 52 1914

Laboulbeniales

Laboulbeniaceae

Amphoropsis Platysthethus 10

Aposporella 18

Autophagomyces 48

Cantharosphaeria 18

CochHomyces 8

Coreomycetopsis 18

Crytandromyces 48

Cucujomyces Monicomyces 8

Diandromyces 48

Eudimeromj'ces 48

Endosporella 18

Entomocosma Anformortidea 10

Helodiomyces ^/ 54

Ilytheomyces Ji 48

Laboulbeniella '. 8

Laboulbeniopsis 18

Mimeromyces Sphaleromyces 48

Myriapodophila llcrpomyccs 10

Nyctcroinyces 48

Pselaphidomyces Stichomyces 8

Scaphidiomyces 48

A PRESENTATION OF NEW GENERA OF FUNGI 5/

Scelophoromyces 48

Stephanomyces Monicomyccs 8

Sj-nandromyces 48

S>'naptomyces 48

Tengandromyces 48

Termitaria 18

Tetrandromyces 48

Thaxteriola 10

Trenomyces Dimeromyces 54

BASIDIOMYCETKS

USTILAGINALES

Ustilaginaceae

Anthracocystis Ustilago 63 15 53 1912

Mycocoscoma 63 15 50 1912

Uredinales

Melampsoraceae

Bottyorhiza '. . . Endophyllum 3 4 47 1917

Crossopsora 7 16 243 1918

Endophylloides Endophyllum 3 4 50 1917

Oliveo 41 9 61 1917

Pucciniaceae

Alevomyces Uromyces 5 28 190 1914

Anthomycetella '. 7 14 353 1916

Calidion 7 16 42 1918

Caronotelium 7 19 174 1921

Cephalotelium 7 19 165 1921

Chrysocelis 39 5 542

Clenoderma 4 17 103 1919

Cleptomyces Calliospora 18 65 464 1919

Ctenoderma 7 17 102 1919

Cystopsora 7 8 448 1910

Cystotelium Longia 7 19 165 1921

Desmella 7 16 241 1918

Dichlamys 7 19 105 1919

Graveola 7 19 173 1921

G>Tnnotelium 7 19 170 1921

Haplaravenelia 7 19 165 1921

Haplopyxis 7 17 105 1919

Kunkelia 18 63 504 1917

Linkiella 7 19 173 1921

Longia 7 19 165 1921

Miyagia 7 11 107 1913

Nielsenia 7 19 171 1921

Nothoravenelia Ravenelia 7 8 310 1910

NyssopsoreDa 7 19 169 1921

Ontotelium '. 7 19 174 1921

Oplophora 7 19 170 1921

58 O, A. PLUNKETT, P. A. YOUNG, AND RUTH W. RYAN

Peristemma 7

Phragmotelium 7

Pleomeris Nielsenia 7

Sclerotelium 7

Teloconia 7

Trachizsporella 7

Triactella 7

Tricella Calliospora 41

Trocochodium 7

Triphragmiopsis 54

Uredinales Imperfecti

Argom^'ces 43

Xenostele 7

Auriculariales

Auriculariaceae

Hoehnelomyces Pilacrella 16 37 514 1919

Tremellales

Tremellaceae

Gloeosoma 7 18 51 1920

Phaeotremella 59 5 376 1911

Dacryomycetales

Dacr\-omycetaceae

Dacrj'opsella ■ 53 124 49 1915

Agaricales

Thelephoraceae

Duportella Stereum 47

Jaapia 7

Peniophorina Peniophora .53

Hydnaceae

Gloiothele 7

H3-dnodon Hydnum 41

Polj^oraceae

Echidnodia 54

Ermeria Daedalca 32

Pseudopolj'porus 41

Xanthoporia 41

Agaricaccac

Amanitella \manita 7

Catathelasma 18

Chloroph}-llum 43

Chlorosperma 41

Copelandia 32

Lentodicllum 41

Micropsalliota Agaricus 53

A PRESENTATION OF NEW GENERA OF FUNGI 59

Plicaturella 43

Pol}'ozellus 33

Rhodopaxillus Tricholoma 7

Phallales

Phallaceae

Protophallus Phallogaster 41 2 25 1910

Clathraceae

Pharus Lysurus 83 7 60 1919

Hymenogastrales

Hysterangiaceae

Jaczerwskia 37

Phaeocryptopus Cryptopus 54

Hymenogastraceae

Stephanospora Hydnangium 54

Sclerodermataceae

Neosaccardia Scleroderma 75 56 6 1921

Lycoperdales

Lycoperdaceae

Geasteroides Geasteropsis 41 9 271 1917

Lycoperdellon Lycoperdon 20 11 92 1912

FUNGI IMPERFECTI

Sphaeropsidales

Sphaerioidaceae

Amphiciliella 32

Amphorula Kellermania 43

Amylirosa Ephclidium . 10

Angiopomopsis 53

Bakerophoma 7

Botryella 7

Botryogene 7

Botrj'osphaerostroma Diplodia 32

Calopactis 7

Camarographiuni 16

Caudosporella Harknessia 53

Ceratophoma Sphaeronema 32

Ceratopycnis Hendersonia 53

Chaetocytostroma Fusicoccum 7

Chondropodiella 32

Cladochaete 7

Collonaemella 53

Columnothyrium 16

Cornucopiella 53

Cr>'ptorhynchella Sphaerographium 53

Crytosphaerella ' 53

60 O. A. PLUNKETT, P. A. YOUNG, AND RUTH W. RYAN

Cytonaema Cytospora 53

Cytophoma Cytospora 53

Cytoplacosphaeria 7

Cytosphaera 7

Cytostagonospora Stagonospora 7

Cytotriplospora Cytospora 59

Dasypyrena 8

Dasysticta 8

Dearnessia Stagonospora 32

Diplodothiorella Dothiorella 7

Diploplacosphaeria Thoracella 32

Dothorellina Dothiorella 16

Dothisphaeropsis 53

Ectosticta 8

Endogleoea Sirostomella 66

Fumagospora 4

Gamonaemella Gamospora 49

Haplosporidium Pyrenochaete? 8

Hemidothis 7

Hendersonina 38

Herpotrichopsis 53

Jahniella 7

Lasiostroma 54

Leptophoma 53

Lichenophoma Dendrophoma 32

Linochora 53

Linchorella Linchora 7

Malachodermis 32

Macrophomella Macrophoma 7

Mastigosporella Harknissia 53

Myriellina 53

Myxofusicoccum Fusicoccum 7

Neohendersonia Hendersonia 7

Neoplacosphaeria Placosphaeria 7

Neosphaeropsis Sphaeropsis 7

Phacidiopycnis 67

Phaeocytostroma Cytospora 7

Phellostroma 47

Phyllostictina 7

Placodiplodia 16

Placonaemina 7

Placophomopsis Phomopsis 34

Placothyrium Cytosporina 16

PlectonaemcUa 53

Pleocouturea ^^^ 4

Pleosi)haeropsis ' Sphaeropsis 7

Pleuronaema Sphacronema M

Pleurophoma Dendrophoma 53

Plcurophomella Dothiorella 53

Polyopeus Phoma 43

Pseudodiplodia Diplodia 10

Pseudohaplosporeila 10

A PRESENTATION OF NEW GENERA OF FUNGI 61

Pycnis Sclcrophoma 53

Pycnosporium Cicinnobolus 26

Pyrenochaetina 7

Rhodoseptoria 54

Sarcophoma Sclerophoma 53

Sclerochaetella Plendomus 32

Sclerophomella Phoma 32

Sclerophomina Plioma 32

Scleropycnis 7

Sclerosphaeropsis Sphaeropsis 5

Sclerostagonospora Stagonospora 32

Sclerotheca Camarosporium 57

Scleroth>'rium Sclerophoma 32

Scolecosporella Hendersonia 7

Septoriopsis Septoria 41

Sirophoma Phoma 32

Sirospenna 72

Sirosphaera 47

Sirostomella 53

Sphaeriostromella Phomopsis 16

Sphaeronaemina Sphaeronema 32

Sphaerothyrium Sclerophoma 16

Steganopycnis 7

Stenocarpella 7

Subulariella Sphaerographium 53

Traversoa Sphaeropsis 7

Trotteria 87

\"ermiculariopsis Vermicularia 20

Verrucaster 2

Nectrioidaceae

Blennoriopsis Sirothyrella 7

Cyanophomella 32

Cyanochyta 53

Dothiorina 53

Gyrostroma 54

Leptodermella 66

Mycorhyuchella 32

Pycnidiella 53

PIenoz3lhia ' 7

Scleropycnium 58

Sirocyphella 53

Stylonectria (Fam. doubtful) 53

Stylonectriella 53

Leptostromiitaceae

Chaetopeltiopsis Chactothyrium 19

Didymochora 32

Diedickea Pycnothyriaceae 7

Discosiella Discosia 36

Discothecium 7

Ichnostroma ■ 47

62 O. A. PLUNKETT, P. A. YOUNG, AND RUTH W. RYAN

Khekia 32

Lasiothyrium 47

Leptothyrina Leptothyrium 53

Massalongina Leptostroma 16

Peltaster Asterostomula 7

Phaeolabrella Labrella 8

Pirostomella 7

Pleurothyrium Leptostromella 16

Pycnothyrium 7

Rhabdothyrella 53

Rhabdothyrium 53

Rhizothyrium Septothyrella 54

Sirothyriella 53

Sirothyrium 7

Sphaerothyrium Leptostroma? 16

Thyriostroma 7

Trachythyriolum 77

Pachystromaceae

Hypodermina 53

Microdiscula 53

Pachydiscula Scleropycnis 66

Rhabdostromella 53

Tryblidiopycnis 53

Xenostroma 53

Excipulaceae

Acleistia 59

Acrosporium 16

Bacterexcipula 32

Chaetodiscula 32

Desmopatella 16

Dinemasporiella. . . , Dinemasporium 32

Excipuella Excipula 53

Exotrichum 7

Falcispora 32

Phaeopolynema Excipula 8

Psalidosperma 7

Pseudolachnea Pseudopatella 7

Ramulariospora 5

Stauronema 7

Stictepatella 32

Melanconiales

Melanconiaceae

Basilocula 7

Cheiroconium 53

Colletotrichella Labrella 53

Cryptosporiopsis 32

Didmosporina Chalara 53

Discosporium 66

Elaedemia 7

A PRESP.XTATION OF NEW GENERA OF FUNGI 63

Gloesporidiella Gloeosporium 32

Gloeosporidina Gloeosporidiiim 7

Heteroceras 7

Marssoniella Marssonina 53

Myrioconium 7

Stegosphaeria Marsonia 7

Titaespora 7

Titaesporina 7

MOXILIALES

Moniliaceae

Amblyosporiopsis Amblyosporium 49

Beauveria Spicaria 21

Cristulariella Cristularia 53

Gemmophora 16

Helicodendron 44

Hormactinia 32

Oosporoidea Oospora 41

Pachybasidiella .' 7

Polymorphomyces 71

Ramulispora Ramularia 88

Sporoclenia 12

Triposporina . . . 53

V'erticilliastrum Verticilliopsis 22

Dematiaceae

Casaresia 80

Ceratosporella 16

Chalaropsis 55

Cheiropodium Clasterosporium 7

Columnophora Chalara 7

Cystodendron 7

Dendryphiella 7

Dichotomella 7

Didymolrichum 53

Endophragmia 54

Eriomenella 44

Harziella Cladosporium. 84

Hormisciopsis Hormiscium 41

Lacellina 7

]\Ielanopsamella Conytrichium 16

Microbasidium Haplobasidium 7

Muiaria Macrosporium 18

Muiogone Sporodesmium 18

Phialophora 41

Piricauda Stigmella 7

Pleurothecium Acrothecium 16

Septoidium 6

Sirosporium Macrosporium 32

Stigmopsis Stigmella 7

Toruloidea Torula 41

64 O. A. PLUNKETT, P. A. YOUNG, AND RUTH W. RYAN

Slilbaceae

Calostilbella 16

Cladographium 44

Coeleographium 24

Coremiella 7

Melanographium Sporocybe 7

Phaeostilbella Stilbella 16

Sporostachys 87

Stilbodendron 7

Synnematium Hirsutella 41

Tuberculariaceae

Amphichaete Chaetospermum 53

Anomomyces 16

Cheiromycella 55

Clathrococcum Epicoccum 53

Discofusarium 59

Exosporella 53

Leucodochium 7

Marcosia 7

Periolopsis 9

Petrakia 7

Phanerocorynella 16

Sigmatomyces 7

Sirodochiella 7

Thyrostroma Epicoccum 53

Tuberculariella 66

Verticilliodochium Tubercularia 7

Xiphomyces 7

The following fungi are of UNKNOWN AFFINITY.

Graphiolaceae

Stylina 7 18 192 1920

Coccoidaceae

Coccodiella 19 25 222 191 1

ASCOMYCETES

Haplostroma 7

Konenia 19

Melanomyces Pseudoparodia 7

Miyakeamyces 19

Solanclia 40

FUNGI IMPERFECTI

Chlamydosporium ii

Gloeodes 60

Menezesia 20

Nothospora ' 33

Saprophorum (Hyphomycete) 72

Sirostomella 53

Spirospora 54

A PRESENTATION OF NEW GENERA OF FUNGI 65

StenocarpcUa 7

Trichodiscula 51

No knowfedge as to position

Alichora 16

Amphoromorpha 18

Candelospora 50

Canthransiopsis 18

Cryptoacsus 56

Diploospora 34

Ephelidium .10

Fusicladiella Carlia 16

Heptasporium 63

Mitopeltis 77

Neofabraea Mollisiaceae 17

Parendomyces 25

Peltomyces 27

Pericystis 9

Phaeocryptopus Cryptopus 79

Plenophysa 7

Rachisia 30

Rachodiella 55

Xenopeltis 7

Mycelia Sterilia

Graillomyces 18

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65. Wochenschr. fiir Brauerie.

66. Zeitschrift fiir Garungsphysiologie.

67. Zeitschrift fiir Pflanzenkrankheilen.

68. Botanisk Tidsskrift.

69. Phytopaihology.

70. Vcr. Zool.-Bol. Gcsellschaft Wein.

71. Revue Generalc Botanique.

72. Englcr's Botanisches Jahresbericht.

73. Sitzenberichte Kais. Bohn. Ges. Wiss. Math, Classe.

74. BuUetino dell'otro Botanico. di Napoli.

A PRESENTATION OF NEW GENERA OF FUNGI 67

75. Atti della Reale Accademia delle Scienze di Torino.

76. Bui. Dep't. Landb. Suriname.

77. Boletin de la Accademia Nac. de Ciencias en Cordoba.

80. Bol. Real. Soc. Esp. Hist. Nat.

82. Journal of Agricultural Research.

83. Ann. Roy. Bot. Gar. Perademja.

84. Atti Reale .\ccad. Lincei.

85. Nova -Acta Reg. Soc. Sci. Upsal.

86. South African Journal of Science.

87. .\tti dell'Acad. Veneto-Trentino-Istriana.

88. Bui. S. Manchuria R. R. Co., .Agr. E.xp. Sta. (Kunchu-ling, Manchuria.)

89. Resultados de la Primera E.xpedicion a Tierra del Fuego (1921) (Univ. Nac. Buenos

Aires.)

DEPARTMENT OF METHODS, REVIEWS, ABSTRACTS,

AND BRIEFER ARTICLES

HEMISTOMUM CONFUSUM, A HOMONYM

By

John E. Guberlet

Parasitologist, Oklahoma Agricultural Experiment Station

Dr. Maurice C. Hall recently called the attention of the writer to the

use of a homonym in connection with the description and naming of a

new species of holostome, Hemistomum confiisum Guberlet, 1922 (Jour.

Parasit., 9:6-14). The specific name conjusum is a homonym of a species

described by Krause, 1914, (Ztschr. f. Wissensch. Zool., Leipz. & Berl.,

112:93-238), whose work on holostomes was unknown to me during my

investigations. Therefore, a new specific name must be submitted for

my species, for which I propose indistincta in place of confusum.

Hall and Wigdor, 1918 (Jour. Am. Vet. Med. Assoc, 53:616-626),

whose work was overlooked by me, were also apparently unaware of

Krause's work but arrived at the same conclusion as Krause relative to

the nomenclature of the genus Hemistomum Diesing, 1850. In both works

it is shown that the name Hemistomum is not in good standing and that

the name Alaria Schrank, 1788, is the correct generic term with alala

Goeze, 1782, as the specific name for the type species. Therefore, according

to priority, the combination should be Alaria aJata (Goeze, 1782) Krause,

1914.

In view of the foregoing and following the suggestion of Hall by com-

munication, I wish to substitute the generic name Alaria, in place of

Hemistomum, for the holostomes recently described by me (1922), i.e.

H. gavium and H. indistincta. Hence the substitution will change the

names to Alaria gavia and .4. indistincta.

68

THE USE OF SODIUM SILICATE AS A MOUNTING

MEDIUM

By

Charles W. Creaser and William J. Clench

Museum of Zoology, Universily of Michigan

In preparing slides for use in two widely different fields the writers

have had occasion to use a solution of sodium silicate (water-glass) as a

mounting medium. Several investigators have used this substance but

there seems to be no literature on the subject. Mr. W. F. Clapp of Cam-

bridge, Massachusetts, has used it in mounting the radulae of certain

mollusks. Through a knowledge of his use of sodium silicate as a mounting

medium, the present development had its inception. Several investigators

have inquired about the methods best adapted for working with it, and

its availability for their problems. Our experience with this substance is

made known here.

Sodium silicate is soluble in water, and basic in reaction, therefore, it

can only be used with basic stains. Fish scales may be mounted as total

objects from water without staining. In mounting radulae of certain

fresh water mollusks, basic fuchsin has given very good results.

For the stock of sodium silicate, we have used that sold by the ordinary

drug store for preservation of eggs. We find it as good as any of the more

carefully prepared solutions. Care must be used in the selection of the

stock since this substance not uncommonly contains a fine white precipitate

in suspension. This kind of material produces a mount with a milky or

clouded appearance. Sodium silicate may be kept in the ordinary balsam

bottle, but it should not be stored in glass-stoppered bottles.

The ordinary water solution of sodium silicate, in its commercial

form, may be used as described here. It has, however, certain defects

that render the slides unfit for use after approximately three months.

We have used it to prepare mounts that are to be studied in a vertical

position. By the use of this material, solid, durable mounts may be

produced which do not melt or run when heated and in a very short time

may be used in a vertical position. It is for this specific purpose that

we have found sodium silicate very useful. Objects mounted in glycerine-

jelly will not stay in position under the influence of heat and gravity

until after they have been prepared for some time. Glycerine jelly and

similar media have a tendency to clear, but it is necessary to avoid this

effect in fish scales and moUusk radulae. In field work where it is difficult

69

70 CHARLES W. CREASER AND WILLIAM J. CLENCH

to make mounts with a medium that must be heated the use of a cold

solution of sodium silicate has much in its favor. Optically, these slides

are excellent in revealing the characters of fish scales.

It has been found that mounts made, of the commercial solution of

sodium silicate are not permanent since this medium will crystallize in

three or four months. However, this difficulty seems to have been over-

come by the use of glycerine in connection with the sodium silicate. Our

slides, made according to this revised formula, have not been kept long

enough to show what the ultimate outcome will be, but after several

months they show no tendency to crystallize.

The fish scales are placed in water, cleaned, and allowed to soak for

a time. They are then transferred to clean water or to dilute solutions

of glycerine or of sodium silicate. Mounting from either water or glycerine

into sodium silicate has been found very satisfactory. Care must be used

that the object is free from all alcohol as a trace of this will cause a white

precipitate to form over the object when it is placed in the sodium silicate.

It has been our practice to put the sodium silicate solution on the slide

by the use of a solid glass rod or to pour it direct from a small bottle. The

medium should be spread over most of the area to be covered by the

cover glass. The objects are then transferred from the water or the dilute

solutions of water-glass or glycerine before they dry and placed in the

desired position on the slide. Objects may be placed on the slide lirst in a

small drop of water, the water-glass and the cover glass being put on

afterwards.

Since the water solution of sodium silicate quickly forms a tough

film over the exposed surface much care and speed must be used in placing

on the cover glass, in order to prevent air bubbles and to insure the spread-

ing of the medium to all parts of the area under the cover glass.

The slides are allowed to set. Air bubbles will work their way out if

the slide is at a very slight angle during this time. These slides will set

in two or three days and can then be used in a vertical position and the

objects will not be displaced by heat or gravity.

Cleaned radulae are transferred directly from water to a water soluble,

basic stain and then returned to water where the excess stain is removed.

The" are then mounted directly in a drop of water-glass in the same

manner as are the fish scales. Ringing the slides in the ordinary manner

will insure greater permanence in these mounts.

M Glycerine sodium silicate may be prepared as follows: To a solution

of commercial sodium silicate, slightly diluted glycerine is added and the

liquids mixed by shaking. The proportions may vary but the best results

are obtained by using twelve parts of water-glass to one part of glycerine.

If the mixture does not go into a homogeneous solution a little water

should be added. This medium does not set as rapidly as the sodium sili-

SODIUM SILICATE AS A iMOUNTING MEDIUM 71

cate solution and is therefore easier to handle. Objects may be mounted

in it in the manner described above. An excess of medium may be removed

with warm water and discarded slides placed in water may be easily

cleaned after a few hours. Slides prepared from this medium are excellent

for the study of fish scales and mollusk radulae. They seem to be quite

permanent.

NEW POCKET DISSECTING MICROSCOPE

E. G. Campbell of Purdue University has described and illustrated

(Science, 192v3, 57:179-180) a simple, efficient instrument for the examina-

tion of small objects in the field. Focusing, rotation of the object, and

dissection may all be performed simultaneously and with ease as the

microscope is held in the hands of the observer. The pocket adjustment

of the instrument provides not only for concealment and protection of

the working parts but also storage space for dissecting instruments.

Change from pocket to working adjustment is simple and quickly accom-

plished. Two figures show clearly the details of construction of this

instrument.

THE PHYSIOLOGY OF REPRODUCTION, by F. H. A. Marshall.

Second revised edition; 770 pages, 189 illustrations. Longmans,

Green and Co., London and New York, 1922. Price $12.00.

Since Dr. Marshall's "Physiology of Reproduction" first appeared it

has been the standard and very useful text and reference book in its

field. The excellent arrangement of its materials, its brevity of statement,

and the clearness and cogency of its summaries have made it one of the

most satisfying and dependable reviews of knowledge in any field of

comparative biology. The manner of handling the references to original

papers is adequate, and gives a historical view without being overwhelming.

The new edition loses none of the admirable qualities of the first and

brings the survey up to date by recording the progress of the last twelve

years. Aside from such detailed revision certain large topics have been

rewritten and elaborated. Some of these are: The Biochemistry of th^

sexual organs; Changes in the maternal organism during pregnancy;

Fertilization; Internal secretory functions of the reproductive glands;

Sex-determination, and the causes of birth. A special discussion is included

of Child's theory of rejuvenescence and senescence as these bear upon

certain types of reproductive and life cycles not so well explained by the

theory of the continuity of germplasm and the segregation of germ cells.

Aside from its indispensable use to all teachers and students of biology,

zoology, and physiology, the book is of high value to gynecologists, veterina-

rians, and animal breeders. Subject and author's indices are excellent; the

printing, illustrating, and mechanical appearance are worthy of the matter.

T. W. Galloway

72

MARY ALLARD ROOTH

September, 8, 1843 September, 15, 1922

Ne^^r has a family motto been typified so truly as in the life of Mary

Allard Booth. "What I hope to accomplish I shall accomplish" seems to

have been the daily inspiration which aided her, in spite of manv obstacles,

to the highest achievement in the labors undertaken.

Born at Longmeadow, Massachusetts, on September 8th, 1843 the

years of her childhood and young womanhood were a record of ill health

and affliction, and during her early education in the local schools, much of

tiie work and play of ordinary youth were denied her.

In speaking of her family, Miss Booth said, "My mother's people were

a literary family, the Bartons of Vermont, of whom Clara Barton, Presi-

dent of the Red Cross, was one. From my father I inherited mv love of

science.

During a visit to the shores of Long Island Miss Booth's attention was

first directed to the interesting forms of aquatic life. While seated in her

wheel chair she watched a woman near bv who was intent upon gathering

sea weeds. The woman was Miss Mary Halliday of Brooklyn, and during

the chance acquaintance which the occasion offered. Miss Halliday took

pleasure m explaining to the invalid girl the wonderful structure and life

history of some of the marine algae.

This incident seemed to be just what was needful to awaken the interest

which her father had long sought to arouse, and with his help and knowl-

edge at her constant disposal she began the work which was to fill her long

life with an unfailing enthusiasm and which bv painstaking application

placed her at last in the foremost rank of scientific workers.

In 1877 she purchased her first compound microscope and step bv step

advanced into the wonderland which it disclosed. The delicate manipu-

lation necessary in the work, the skill in preparation and mounting of

specimens were the result of long and patient endeavor due to love of the

work. No object in the field of nature was too minute for careful study

VA/hen the wonders of this new world were thus revealed to her. Miss Booth

sought some method by which she could make this microcosmos intelli-

gible to others and there the camera seemed to fill the required need.

With microscope and camera, then, she was able to make of vast impor-

tance to humanity the line of study so assiduouslv pursued. When the

invasion of bubonic plague seemed imminent in this country Surgeon

73

74 BESSIE PERRAULT TITUS

General Blue turned to Miss Booth with a request for photomicrographs

of the plague fleas infesting rats in San Francisco, and these photographs

were used in a nation-wide lecture campaign for the extermination of the

plague. Not only this Government but those of France and Fngland as

M AKY A. HocJTII

well sought her aid in the tight against such subtle enemies. Many infini-

tesimal creatures were sent on the long journey to her laboratory in Spring-

field, Massachusetts, there to be photographed and studied scientifically.

MARY ALLARD BOOTH 75

All the steps in the painstaking process from the preparation of the

object, proper staining to emphasize structural detail, delicate mounting

on microscopical slide, photographing, developing the negative and making

the prints, were performed by this patient worker, and each picture was a

masterpiece of its kind. And in the midst of all this patient endeavor and

accomplishment, Miss Booth found time to make for herself an honored

place in the lecture field and editorship of several scientific journals.

The great esteem in which she was held at home and abroad is evidenced

by the honors conferred upon her. Few women have shared with her the

distinction of fellowship in the American Association for the Advancement

of Science, and she was likewise one of the few women elected a fellow of

the Royal Microscopical Society of London. As a higher reward for her

humanitarian services the privilege was given her to continue her activity

in full strength and vigor of mind up to the very close of her life.

Her latest photographs were those of a family of busy little squirrels

who made their home in a neighboring tree and who had learned to know

and love the good friend who cared for them so faithfully. Indeed, as was

a fitting close for such a lover of nature her last act on earth was the carry-

ing of the evening meal to these little creatures; and in the great out-of-

doors, at the close of a September day, surrounded by these appreciative

friends, she passed into the great beyond.

Bessie Perrault Titus

PROCEEDINGS OF THE AMERICAN MICROSCOPICAL

SOCIETY

Minutes of the Boston Meeting

The 41st annual meeting of the American Microscopical Society was held in affiliation

with the American Association for the Advancement of Science at Boston, Mass., December

28, 1922.

President N. A. Cobb presided at this meeting.

The report of the Treasurer for the year 1922 was read by the Secretary and was referred

to an Auditing Committee composed of Professors F. H. Krecker and J. W. Kostir.

The report of the Custodian was read by the Secretary and was referred to an auditing

committee composed of Messrs. Edw. Pennock and F. E. Ives.

The meeting voted to send hearty congratulations to the Custodian on the growth of

the Spencer-Tolles Fund.

The Secretary presented a report on the affairs of his office, this report covering the

previous three years.

The following officers were nominated and elected: President, Professor Chancey

Juday, University of Wisconsin, Madison, Wis.; First Vice-President, Dr. B. H. Ransom,

Bureau of Animal Industry, Washington, D. C; Second Vice-President, Dr. W. W. Cort,

Johns Hopkins University, Baltimore, Md.; Secretary, Professor Paul S. Welch, University

of Michigan, Ann Arbor, Mich.; Treasurer, Dr. Wm. F. Henderson (for two years), University

of Pittsburgh, Pittsburgh, Pa.

Professor Geo. R. La Rue, University of Michigan; Professor Z. P. Metcalf, North

Carolina State College of Agriculture and Engineering, and Professor E. M. Gilbert, Univer-

sity of Wisconsin, were chosen as the elective members of the Executive Committee for 1923.

Dr. B. H. Ransom, Bureau of Animal Industry, was chosen as a representative of the

Society on the Council of the American Association for the Advancement of Science.

Dr. N. A. Cobb, Bureau of Plant Industry, was appointed as a member of the Spencer-

Tolles Fund Committee.

Adjourned.

Paul S. Welch, Secretary.

CUSTODIAN'S REPORT FOR THE YEAR 1922

Spencer-Tolles Fund

Balance reported for the yeaj 1921 S9104 . 56

Interest on Bonds 250.00

Dividends Penna. R. R. Co 96.75

*Build'g. & Loan Ass'n 144. 16 490.91

9595.47

Less Grant to Wm. P. Hayes 60 00

95,>5 47

Net Increase during the year S4.?0.91

*Estimates, proved correct by letter of B. & L. Ass'n. of Dec. 19-"22. M.P .

76

PROCEEDINGS OF THE AMERICAN MICROSCOPICAL SOCIETY / /

Totals

Receipts

All contributions 802 .03

All sales 1193.38 •

All life memberships 300.00

All interest, dividends, profits 7590 . 06

9885.47

Disbursements

All Grants 310.00

All life membership dues 40.00 350.00 9535.47

Investments

Stock in Keystone State Bldg. & L. Ass'n 2385 .47

Bonds, Rio Grande Junction R'y 5000.00

Stock, 43 shares Penna. R. R. Co 2150.00 9535.47

Life members: (Robert Brown, dec'd.); J. Stanford Brown, Seth Bunker Capp, Harry B.

Duncanson, A. H. Elliott (dec'd.) and John Hately (dec'd.).

Contributions of $50 and over: John Aspinwall, Iron City Microscopical Society, Magnus

Pflaum and Troy Scientific Society.

(signed) Magnus Pflaum

Custodian

Philadelphia, Pa.

Dec. 30, 1922

Philadelphia, Feb. 1st. 1923

Having examined the above account, and the securities on hand as shown therein, we find

jj^em correct.

F. E. Ives

Edward Pennock

ANNUAL REPORT OF THE TREASURER OF THE

AMERICAN MICROSCOPICAL SOCIETY

December 24, 1921 to December 13, 1922

Receipts

Balance on hand, December 24, 1921 $ 592 . 71

Dues received for Volume 40 or before 45 . 00

Dues received for Volume 41 170 . 10

Dues received for Volume 42 252 . 00

Dues received for Volume 43 2 . 00

Initiation Fees 24 . 00

Subscriptions for Volume 40 or before 3 . 00

Subscriptions for Volume 41 315.02

Subscriptions for V' olume 42 54 . 00

Sales of Transactions, duplicates, back numbers 26.21

Advertising, Volume 39 10 . 00

Advertising, Volume 40 250.00

Authors, for preparation of plates 34 . 30

Grant, from Spencer-Tolles Fund 60 . 00

Sundries , -50

Total S1838.84

78 PROCEEDINGS OF THE AxMERICAN MICROSCOPICAL SOCIETY

Expenditures

Printing Transactions, Volume 40, No. 4 $ 224.02

Printing Transactions, Volume 41, No. 1 257 . 16

Printing Transactions, Volume 41, No. 2 256. 77

Printing Transactions, Volume 41, No. 3 200.05

Postage and Express for Secretary 21 . 00

Postage and Express for Treasurer 13 .00

Office expenses of Secretary 47 . 82

Ofifice expenses of Treasurer 28 . 65

Secretary, trip to Toronto 39 . 89

Balance on hand 750 . 48

Total $1838.84

December 13, 1922.

\V. V. Henderson', Treasurer.

Report of the Auditing Committee of the American Microscopical Society

The accounts of W. F. Henderson, Treasurer of the American Microscopical Society,

for the period beginning December 24, 1921, and ending Dec. 13, 1922, have been examined

by the Auditing Committee and have been found to be correct.

Respectfully submitted,

W. J. KOSTIR

Feb. 28, 1923. F. H. Krecker

TRANSACTIONS

OF THE

American

Microscopical Society

Organized 1878 Incorporated 1891

PUBLISHED QUARTERLY

KY THE SOCIKTV

EDITED BY THE SECRETARY

PAUL S. WELCH

ANN ARMOR, MICHIC.AN

VOLUME XLIl

Number Three

Entered as Second-class Matter August 13, 1918, at the Post-office at ^lenasha,

Wisconsin, under Act of March 3, 1879. Acceptance for mailing at the

special rate of postage provided for in Section 1103, of the

Act of October 3, 1917, authorized Oct. 21, 1918

W^i OlolUsiatc Prcaa

GEORGE BANTA PUBLISHING COMPANY

MENASHA, WISCONSIN

1923

TABLE OF CONTEXTS

For Volume XLTI, Number 2, April, 1923

The Distribution of Frog Parasites of the Douglas Lake Region, Michigan, by Harry C.

Fortner 79

New Records of North American Enchytraeidae, by Paul S. Welch 91

Primitive Microscopes and Some Early Observations, by William A. Locy 95

An Illuminating Device for Microscopes, by William A. Beck 108

Abnormal Specimens of Helodrilus caliginosus, trapezoides (Duges) and Hclodrilus

roseus (Savigny), by Bess R. Green 122

Department of Methods, Reviews, Abstracts and Briefer Articles

A Study of the Stability of Staining Solutions, by F. L. Pickett 129

Modern Microscopy, a Handbook for Beginners and Students, a review by George

R. La Rue 133

TRANSACTIONS

OF

American Microscopical Society

(Published in Quarterly Instalments)

Vol. XLII APRIL, 1923 No. 2

THE DISTRIBUTION OF FROG PARASITES OF THE

DOUGLAS LAKE REGION, MICHIGAN^

By

Harry C. Fortner

University of Vermont

Introduction

During the summers of 1917 and 1919 the writer made a study of the

parasites of frogs and their distribution in the Douglas Lake Region,

Cheboygan County, Michigan. It was thought that a study of the distri-

bution of frog parasites, including detailed statistical data, might present

some interesting as well as valuable information. Three species of frogs

were examined within a limited area, thus furnishing data contributing

to the geographical distribution of frog parasites.

In all, two hundred eight hosts were examined from eleven local

stations (See Map Fig. 1.). The collections were made during the months

of July and August of the two years, 1917 and 1919. Some differences

occurred in the two seasons which are worth noting and these will be

stressed in the discussion. The particular habitats from which hosts

were taken are described below.

Bryant's Bog is a typical bog association with plant life slowly encroach-

ing upon the water. As a matter of fact, very few frogs inhabit this

place. So few frogs were taken there that a lengthy description of the

habitat is not warranted.

Carp Creek is a short, rapid, spring-fed trout stream. The majority

of specimens taken from this habitat came from the roadside among the

grass which grew around two large moss-covered logs. A few were taken

from an abandoned road along the stream where mosses and liverworts

were abundant. On account of the swift current, breeding could take

place here only in the side pools. In 1919, but two specimens were taken

in this locality from the roadside. The vegetation of the abandoned road

was dense and practically no open places existed as in 1917.

Fairy Island furnished but one specimen each season. Here the shore

and bottom consisted mostly of gravel. Sedges were the only vegetation

' Contribution from the University of Michigan Biological Station.

79

80

HARRY C. FORTNER

along shore, but they afiforded resting places for several species of Cole-

optera, Hemiptera, and Hymenoptera, which served as food for the frogs.

Lancaster Lake is a small body of water near Douglas Lake, having

an area of several acres. A small stream, Bessey Creek, connects Lancaster

Lake with Douglas Lake, thus presenting a very easy path of migration

for frogs from one lake to the other. The shore was thickly matted with

Fig. 1. Collecting Stations

sedges. Small bushy willows provided a shaded shelter between the

sedges and the edge of the wooded area. It was here that most specimens

were taken. The soil between the water and the wooded area consisted

mostly of muck. Small depressions were present in which numerous

aquatic insects thrived. There was practically no visible change in this

habitat since 1917.

Maple River, the outlet of Douglas Lake, runs tiirough a comparatively

level stretch of country. Scattered along its course are numerous bayous

FROG PARASITES OF DOUGLAS LAKE 81

affording excellent breeding places for frogs. Sedges and grasses were

very dense and stumps and roots of trees lined the banks. Cat-tail societies

were also present. The bottom was muddy or sandy, and at places stones

and gravel were prevalent. Deep holes and shallows were common.

North Fish Tail Bay (N. F. T. Bay in tables), an arm of Douglas

Lake, presents a habitat consisting of a level sandy shore with a thick

growth of sedges. The bottom consists of sandy marl and muck. A

few potomogetons and some chara are present in the water. Here the

frogs have access to many species of insects and some few snails.

Trout Creek is a small trout stream rising in bogs to the north and

east of Douglas Lake. The vegetation along this stream is varied. Typical

roadside plants and trees are present along the banks of the stream.

Small areas of water lilies occur in quiet stretches of water. Algae are

also present. Grasses overhang the water in many places. At some places

the bottom consists of muck; at others gravel forms the main constituent

of the bed of the stream. The stream is swift in some few places; at

others it is slowly running or even stagnant. Some places are in dense

shade and others in bright sunlight. The depth of the water varies from

a few inches to a foot and a half.

Reese^s Bog is a large Thuja bog on the north shore of Burt Lake.

Conditions in this bog had undergone considerable change between the

years 1917 and 1919. At one place where many specimens were taken

in 1917 moisture was lacking in 1919 and very few frogs were taken.

At another place the grass had become very dense and it was too dry for

a frog habitat. In still another area certain mosses and grasses had

become denser, thus holding back more of the surface drainage which

made the habitat too moist for certain kinds of insects. On slightly

higher ground conditions were more favorable but frogs were not as

abundant as in 1917. The Rana clamitans which were taken here were

found near the water and in secluded and well sheltered spots. Rana

pipiens wanders a considerable distance from the stream in contrast to

Rana clamitans, which is always found near the main body of water.

Sedge Point is situated on the north shore of Douglas Lake, not far

from the western margin of North Fish Tail Bay. Here several pools

are cut off from the lake as a result of wave action. Around these pools

there was a zone of water plants. One pool usually containing water

was devoid of water in 1919, but the soil was somewhat moist. Here

insect life was very abundant. Snails, too, were very numerous, eight

species being present; these no doubt figuring largely in the life histories

of many of the parasites.

South Fish Tail Bay (S. F. T. Bay in tables) is the site of the Biological

Station. Here the shore is sandy. There is a very scant growth of plants

in the water, and but few sedges along the shore. Very few frogs make

this a permanent habitat.

82

HARRY C. rORTNER

Sunny Strand is a sedge-covered beach situated at the north-western

part of Douglas Lake. The water along this shore is very shallow for

fifteen or twenty yards out. On account of the direction of the prevailing

winds this is a sheltered shore. No hosts were taken from this place in

1917, but in 1919 four were taken.

Discussion

Three species of hosts were examined, one hundred seventy-seven

specimens of Rana pipiens, twenty-nine Rana clamitans, and two Rana

cantabrigensis, making a total of two hundred eight. No Rana catesbiana

were seen at any time during the two summers, altho they are recorded

for the region. Of the number collected, seven individuals, all of small

size, seemed to be entirely free from parasites. A total of ten species,

excluding the nematodes, parasitic in frogs were found. Six, however,

was the highest number of species found in any individual host. The

nematodes are in the hands of an expert and their distribution will have

to be reported at a later date.

It might be interesting to note that when frogs were fed daily in

captivity, then examined, no loss of intestinal and urinary bladder para-

sites were noted. Loss occurred, however, where no feeding was done

within 12-24 hours, and the parasites were recovered from the bottom

of the aquarium.

Table No. I shows the number of hosts taken from each habitat in

each of the two seasons.

Table No. I

Number of Hosts Taken from Each Habitat

FROG PARASITES OF DOUGLAS LAKE

83

Table No. II shows the parasites found and their local distribution.

Table No. II

Parasites Found and Habitat Distribution

Octomitus intestinalis Prowazek

Opalina obtrigonoidea Metcalf (An as yet un-

published species)

Nyctotherus cordiformis Stein

Diplodiscus temperatus Stafford

Gorgoderina attenuata Stafford

Pneumoneces medioplexus Stafford

Cephalogonimus americanus Stafford

Clinostomum attenuatum Cort

Proteocephalidae

Pneumoneces similiplexus Stafford

M

a;

►5 pa

n

Pi

(X,

a

3

C/2

The identification of trematodes was confirmed by Dr. W. W. Cort;

the Opalina by Dr. M. M. Metcalf; and the other forms by Dr. G. R.

LaRue.

Octomitus was present in the Rana cantabrigensis from Maple River,

and Opalina and a lung nematode were found in the host from Sedge

Point.

Octomitus and Opalina were present in all localities. Nyctotherus was

not secured in frogs from three habitats, probably due to the small number

of hosts taken. Specimens of Diplodiscus were not taken from five of the

eleven localities. Their absence from collections made at Lancaster

Lake and Sedge Point is not to be explained on the basis of small numbers

of hosts examined since large numbers of hosts were taken from those

localities. Gorgoderina seems to be evenly distributed. The heaviest

infection of this parasite noted was fourteen from a single specimen.

Specimens of Pneumoneces medioplexus were taken from all localities

except North Fish Tail Bay. There is a possibility of its being present

there also, as but eight hosts were examined from that region. The

heaviest infection with this species in any one frog was from the Maple

River habitat, one lung containing thirty-eight, the other thirty-four.

Another host from Sedge Point had thirty-six in one lung and thirty-four

in the other. Pneumoneces similiplexus was taken only in the one locality,

Maple River. Six specimens of Cephalogonimus were taken during the

two summers from, three hosts and two localities. Since these two localities

84 HARRY C. rORTNER

are not far apart it would be possible for hosts to migrate readily from

one of these habitats to the other in a short time, and thus account for

its presence in both places. Clinostomuni attenualum was taken in the same

year, 1917, from but two hosts, which were very heavily infested. Proteo-

cephalidae were obtained from three stations from both Rana pipiens and

Rana clamitans.

Protozoan parasites alone infested the very small frogs which leads

one to believe that the frogs become infested with the metazoan parasites

when they feed on animal life. An exception to this fact, however, is

that Dr. L. R. Cary found Diplodiscus temperatus in tadpoles. Any one

of the protozoans, Opalina, Nyctotherus, and Octomitus, does not seem

to be inconvenienced by the presence of the other forms, as nine hosts

examined contained hundreds of all three species. One of the striking

things about the Protozoa is the fact that Opalinae were found in only

one specimen of Rana clamitans of the twenty-nine examined, while

the percentage of infestation of Rana pipiens with this organism is 61

and 80 for the two summers respectively. Why the infestation differs

in the two species is a question that cannot be answered until more exami-

nations have been made paying particular attention to the other inhabitants

of the habitat which may serve as food and act as carriers. Dr. R. W.

Hegner, in an article published since these notes were written, has pointed

out that while the tadpoles of Rana clamitans are infected with Opalinae

the adults very rarely are infected.

A sufficient number of frogs were not collected at the Sedge Point

locality in 1917 to make a fair comparison with those collected during

1919. Collections made at this habitat might shed some light on the

adult forms of the cercariae present there. Extensive studies have been

and are being made on the cercariae of this habitat. For instance no

D. temperatus were found there, and we might conclude if larger numbers

of hosts were examined and none found that the cercariae of that form

do not exist there.

In all the examinations no Acanthocephali were found. This corrobo-

rates statements of other investigators who have found a very limited

number upon examination of similar hosts in North America.

A comparison of the species of the parasites found in Rana pipiens

and Rana clamitans is made in Table III.

As mentioned before, it is a remarkable fact that Rana clamitans

is so lightly infested with Opalina. Diplodiscus, Pneiimoneces medio plexus,

Cephalogonimus, and at least three species of nematodes were present

in Rana pipiens and not found at all in Rana clamitans. Another striking

comparison is the relatively low infestation of Rana pipiens with Pneiimo-

neces similiplexus and Proteocephalidae as compared with that of Rana

clamitans.

FROG PARASITES OF DOUGLAS LAKE

85

Table No. Ill

Comparison of Infestation of Two Species of Frogs Expressed in Percentages of Infested

Individuals to Entire Number Examined

Rana Rana

pipicus damitans

1917

Rana Rana

pipiens damitans

1919

Octomitus

Opalina

Nyctotherus

Diplodiscus

Gorgoderina

Pneumoneces medioplexus

Pneumoneces similiplexus.

Cephalogonimus

Clinostomum

Proteocephaiidae

30

61

2

30

51

5

1

3

2

0

18

5

15

0

50

0

20

0

10

48

80

22

1

38

30

0.9

0

1

0

22

0

66

0

11

0

As is shown in Table IV there is not enough variation in infection

between the sexes to justify the consideration of each sex separately.

The males and females live under the same conditions and in the same

habitat, and, as would be expected, the percentage of infection between

the two sexes does not differ to any great extent.

Table No. IV

Percentage of Infection of Total Number of Frogs (112) Collected During the Year 1919

57

Females

55

Males

112

Males and Females

Opalina

Nyctotherus

Octomitus

Bladder flukes. . .

Lung flukes

Diplodiscus

Cephalogonimus .

Proteocephaiidae .

70

19

50

36

28

3

1

74

25

52

43

27

0

72

22

51

39.5

27.5

1.5

.5

Comparisons of the percentage of infestation of both sexes of the

same locality also give similar results.

The comparison of the percentage of infection of the hosts of the two

summers in the various collecting places shows but slight variation except

where a habitat was affected by drought or other factors. Variation is

slight particularly where a large number of hosts were taken and can

readily be seen in Table V.

86

HARRY C. FORTNER

Table V

Comparing Percentage of Infestation of Two Summers in the Various Localities

An abundance or lack of food appears to be a factor of considerable

importance. The more a host eats the greater opportunity it has to become

infected. Abundance of food for frogs depends upon abundance of rainfall

and vegetation. Among the stomach contents of the hosts examined were

noted adults of the following orders of Insects: Hemiptera, Orthoptera,

Coleoptera, Lepidoptera, and Hymenoptera. Representatives of the

following groups of Coleoptera were recognized: Chrysomelidae, Scoly-

tidae, Prioninae, and Cicindelidae. Larval forms of Hemiptera, Coleop-

tera, Lepidoptera, Hymenoptera, and Diptera were also found.

FROG PARASITES OF DOUGLAS LAKE

87

Conditions of the habitats at Lancaster Lake and Maple River re-

mained practically the same from 1917 to 1919, and no great differences

were found in the percentage of infection by comparing the two summer's

finds. Pneumoneces medioplexus seems to be the exception here. At

Carp Creek the frogs were not so abundant in 1919 as in 1917, and the

vegetation was very different. With two exceptions, the infestations

of Opalinae and Gorgoderinae in frogs from the Carp Creek habitat, were

markedly different in the two seasons. Many species found in 1917 were

not present in 1919.

Table VI gives for each parasitic species the percentage of the entire

number of frogs infested. As can be seen, the heaviest infestations were

with Octomitus, Opalina, and Gorgoderina.

Table No. VI

Percentage of Infestation of Entire Number of Frogs (208) Examined, 1917 and 1919

Octomitus intestinalis

Opalina obtrigonoidea

Nyctotherus cordiformis ....

Diplodiscus temperatus

Gorgoderina attenuata

Pneumoneces medioplexus . .

Pneumoneces similiplexus . . .

Cephalogonimus americanus.

Clinostomum attenuatum. . .

Proteocephalidae

112

Females

37

58

15

13

47

13

2

0

3

96

Males

44

66

17

10

42

20

2

0

2

0

208

Males and Females

40.5

62

16

11. S

44.5

16.5

2

1

1.5

Seasonal differences probably are due to a variety of factors. The

amount and time of rainfall certainly affects the presence of parasites in a

given locality in any one year.

Table VII gives the amount of rainfall and the temperature previous

to and during the collecting periods of both years.

Table VII

Rainfall and Temperature

88 HARRY C. FORTNER

The amount of rainfall previous to the collecting seasons totals approxi-

mately the same for both years. However, in May 1917 there was an

interval of 16 days of dry weather as compared to a shower every two

or three days in 1919. During the time collections were being made in

1917, 3.47 inches of rainfall was recorded as compared to 1.25 inches

during the collecting period of 1919. Most of the rain that fell during

the 1919 collecting season came near the close of the season in one or two

heavy showers. The small amount of rainfall during the collecting season

of 1919 might account for the low percentage of infection with some forms

as compared with the season of 1917. For instance, hosts taken at Reese's

Bog during 1919 contained no Diplodiscus, and no Cephalogonimus were

found in any hosts examined that year. The percentage in the majority

of instances runs lower for 1919 than for 1917.

Drought seriously affected the food plants of many insects in some

habitats during 1919, and if insects enter into the life history of some

parasitic forms, then this might account for the absence of adult parasites.

The temperature of this region does not vary enough from season to

season to be reckoned as an important factor. Erosion, weathering, and

winds affecting the habitats are, no doubt, slight factors but some that

cannot be entirely ignored.

The writer takes this opportunity to express his appreciation to Dr.

George R. LaRue, under whose supervision this work was done, for his

encouragement and helpful criticism; and to Dr. W. W. Cort and Dr.

M. M. Metcalf for their aid in the identification of certain species.

Summary

1. A study was made of the distribution of the parasites of frogs in

the Douglas Lake Region, Michigan.

2. Two hundred eight hosts from eleven habitats near Douglas Lake

were examined during the months of July and August of the years 1917

and 1919.

3. Three species of hosts were examined, namely, Ratia pipiens,

Rana clamitans, and Rana cantabrigensls.

4. Ten species parasitic in frogs (excluding the nematodes) were

found. Six species was the highest number found in any individual host.

5. Octomitiis, Opalina, and Gorgoderina were present at all stations.

Fourteen individuals of the genus Gorgoderina were taken from one host.

Diplodiscus was not taken from all habitats. Pneiimoncccs medioplexus

seems to be evenly distributed in habitats studied. As a rule infestations

by this species run high, instances of 70 and 72 from one frog being recorded.

But very few hosts contained Pneumoncccs simiUplexns, Cephalogonimus

americanus, Clinostomum attenuatum, and Proteocephalidae.

6. The light infestation of Rana clamitans with Opalina is remarkable

compared to the heavy infestations of Rana pipiens.

FROG PARASITES OF DOUGLAS LAKE 89

7. All stages in the life histories of many of the flukes are not known.

Many cercariae are present in a given locality. By a study of both adult

forms and cercariae of a given station these stages in life histories may be

worked out. If an adult does not happen to be present, then the cercariae

found there, in all probability, are not of that species. Such information

may prove valuable to any one making a study of the cercariae of any given

area or locality. Doubtless the cercariae found at Sedge Point are not a

stage in the life history of Diplodisciis temperatiis as no adults seemed to be

present at that station.

8. No Acanthocephali were found in any hosts examined.

9. No Diplodiscus, Pneumoneces medioplexus, and Cephalogonimus

were found in Rana clamitans, and the infestation of this species with

Pneumoneces similiplexus and Proteocephalidae is comparatively low.

10. Males and females seem to be infested to about the same degree.

11. The food of the hosts, consisting chiefly of insects, the abundance

of which is largely dependent upon rainfall and vegetation, appears to be

a factor affecting the presence or absence of certain parasites. This might

account for the absence of certain species at the Carp Creek habitat in

1919.

12. The amount and time of rainfall appears to affect the presence

of parasites at a given station. The striking examples were that Diplodis-

cus was not found to be present at Reese's Bog and that Cephalogonimus

was not found at all in the vear 1919.

BIBLIOGR.\PHY

Adams, C. C.

1913 Guide to the Study of Animal Ecology. New York, 183 pp.

1919 Migration as a Factor in Evolution: Its Ecological Dynamics, .\merican Natura-

list, 53:55-78.

Gary, L. R.

1909 The Life-History of Diplodiscus temperatus Stafford. Zool. Jahrb., Abt. f. Anat.

u. Ont., 28:595-659.

CORT, W. W.

1912 North American Frog Bladder Flukes. Trans. Amer. Micr. Soc, 31:151-166.

1913 Notes on the Trematode Genus Clinostomum. Trans. Amer. Micr. See., 32:169-

182.

1914 Larval Trematodes from North .American Fresh Water Snails. Jour. Parasitology,

1 :65-84.

1915 Egg Variation in a Trematode Species. Jour. Parasitology, 2:25-26.

1915 North American Frog Lung Flukes. Trans. Amer. Micr. Soc, 34:203-240.

Hegner, Robert W.

1922 The Effects of Changes in Diet on the Incidence, Distribution, and Numbers of

Certain Intestinal Protozoa of Frog and Toad Tadpoles. Jour. Parasitology,

9:51-67.

Holmes, S. J.

1906 Biology of the Frog. New York. 370 pp.

90 HARRY C. FORTNER

Johnston, S. J.

1912 On Some Trematode Parasites of Australian Frogs. Proc. Linn. Soc, N. S. W.,

37:285-362.

Kent, W. Saville,

1880-1881 A Manual of the Infusoria. 3 vols. London.

LaRue, Geo. R.

1914 A Revision of the Cestode Family Proteocephalidae. 111. Biol. Monogr., 1:1-350,

16 pi.

Leidy, J.

1851 Contributions to Helminthology. Proc. Acad. Nat. Sc. Phila., 5:205-209.

Looss, A.

1894 Die Distomen unserer Fische und Frosche. Biblioth. Zool., No. 16, 226 pp.

MiNCHIN, E. A.

1912 An Introduction to the Study of the Protozoa. London.

Pratt, H. S.

1903 Descriptions of Four Distomes. Mark Anniv. Vol., pp. 25-38.

1916 Manual of the Common Invertebrate Animals. Chicago. 737 pp.

Seeley, L. B.

1906 Two Distomes. Biol. Bull., 10:249-254.

Stafford, J.

1905 Trematodes from Canadian Vertebrates. Zool. Anz., 28:681-694.

Stiles, Ch. W. and Hassall, A.

1902 Eleven Miscellaneous Papers on Animal Parasites. Bur. An. Industry, Bull. 35.

Ward, H. B.

1911 The Distribution and Frequence of Animal Parasites and Parasitic Diseases in

N. A. Fresh Water Fish. Trans. Amer. Fish. Soc. pp. 207-241.

Ward, Henry B. and Whipple, Geo. C.

1918 Fresh Water Biology. New York, 1111 pp.

NEW RECORDS OF NORTH AMERICAN

ENCHYTRAEIDAE*

By

Paul S. Welch

Our knowledge of the North American enchytraeid fauna is so meager

that at the present time any definite record is of considerable importance.

Various collections sent to the writer for identification have yielded

specimens representing species previously known only from remote locali-

ties and it has, therefore, seemed desirable to make available those records

which modify so strikingly present impressions as to the range of the

species involved.

Marionina forbesae Smith and Welch

During the month of August, 1921, Mr. R. L. Mayhew collected

sexually mature aquatic enchytraeids from a small pond near the shore

of Burt Lake, Michigan. Preparations resulting from a preliminary

examination were subsequently transmitted to the writer for identification.

Three specimens in the form of serial sections and three as whole mounts,

all prepared by Mr. Mayhew, have been the basis of the work, although

alcoholic material was also available.

Identity. — These enchytraeids exhibit characters which agree exactly

with those described for Marionina forbesae Smith and Welch (1913), except

that (a) in the Michigan specimens the dorsal blood vessel arises in the

posterior part of XIV or the anterior part of XV, instead of in XIII as

in the type, and (b) that small scattering unicellular glands occur at the

ectal opening and along the surface of the spermathecal duct, such glands

being absent in the specimens from Illinois. The writer has not felt

justified in regarding these differences as representing more than variations

within the species.

Mr. Mayhew's records contain no mention of the definitely arranged

superficial spots reported in the original description of the species. Since

alcohol causes these spots to disappear their absence in the Michigan

material may be due to the preservative.

Delphy (1919; 1921) holds that the distinction between Marionina

and ^^Pachydrilus" is not valid. Smith and Welch (1913) had already

noted the close similarity between the two genera. However, pending

more extensive study, the writer has followed the older practice.

Previous record. — M arionina forbesae was originally described from

five sexually mature specimens found in the bottom mud and settlings

of the waterworks reservoir at Urbana, Illinois, in October and November,

1895. This constitutes the only previous record.

* Contribution from the University of Michigan Biological Station, and from the

Zoological Laboratory of the University of Michigan.

91

92 PAUL S. WELCH

Habitat. — The specimens on which this identification is based were

collected from masses of algae growing upon partly submerged boards in

a pond, known as Fontinalis Run, on the northeast shore of Burt Lake,

Michigan, about three miles from the University of Michigan Biological

Station. This pond is merely an expanded end of a stream surrounded

by swamp conditions and opening into Burt Lake through a narrow,

shallow passage. A profusion of invertebrate animals and aquatic plants,

large quantities of decaying organic matter on the bottom, floating wood

bearing masses of algae, very slow current, protection from surface distur-

bances and complete absence of artificial influences are outstanding features

of this habitat.

Fridericia bulbosa (Rosa)

\. A collection made at Mound City, Kansas, July 9, 1914, contained

sexually mature enchytraeids six of which were studied in detail and

found to be typical forms of Fridericia bulbosa. These worms were col-

lected from decaying roots of alfalfa, under conditions which indicated

an indigenous- species.

2. A collection made at Emporia, Kansas, on June 22, 1921, and

sent to the writer by Professor R. C. Smith, Kansas State Agricultural

College, contained sexually mature specimens of Fridericia bulbosa. As in

the previous case, these worms were found in connection with the dead

or dying roots of alfalfa and apparently represent an indigenous species.

3. Sexually mature material of an enchytraeid was found by an in-

spector of the Federal Horticultural Board, on March 17, 1917, in the

soil around the roots of citrus plants growing in the plant quarantine

greenhouse at Washington, D.C. The specimens are clearly Fridericia

bulbosa. The original source of these worms is uncertain. Mr. E. R. Sass-

cer of the Federal Horticultural Board, who sent the specimens, stated

that the plants were originally received from the plant introduction

garden of the Office of Foreign Seed and Plant Introduction at Yarrow,

Maryland and "in all probability, the soil used comes from that locality."

Considering the ease with which these enchytraeids are transported in

soil about the roots of plants the original stock may have been imported

from a foreign locality. However, the finding of representatives of the

same species in central United States under conditions not indicative of

foreign importation suggests the possibility that the Maryland material

may also be indigenous,

Penial bulb. — Specimens from the three above mentioned collections

all agree in the structure of the penial bulb. This organ is of the lumbri-

cillid type in all respects. The body of the bulb is composed of cells of

one kind only, their nucleated portions being near the periphery. A very

few nuclei appear irregularly in the central region. Stephenson (1911,

NEW RECORDS OF ENCHYTRAEIDAE 93

p. 63; PL II, fig. 17) mentions and figures the penial bulb in specimens

from the littoral region of the Clyde and as nearly as can be judged from

the very brief description and the small figure there is complete agreement

with the American specimens.

Chylus cells. — In the Maryland material the chylus cells occur

in XIV-XVI, while in specimens from both of the Kansas collections the

chylus cells begin in XIII and appear to end in XV.

Previous North American Records. — Moore (1895, pp. 343-344) described

a new species under the name Fridericia parva from material collected

in the vicinity of Philadelphia, Pa. Michaelsen (1900, p. 96) regarded

F. parva as a synomym of F. bulbosa and more recent studies indicate the

correctness of this view. If, then, the Philadelphia material be regarded

as F. bulbosa, it constitutes the first and only North American record in

the literature.

Fridericia agilis Smith

Through the courtesy of the Illinois Natural History Survey the

writer had the opportunity to study some enchytraeids collected in the

Sangamon River bottoms near Kilburn, Illinois. These worms were

found about the roots of winter killed wheat in dark soil having a rather

high moisture content and were reported as occurring in considerable

abundance in region where the collections were made. About forty speci-

mens were collected on April 5, 1912, of which thirty were sexually mature.

The color of the living specimens was, in many cases, nearly white

with a slight tinge of flesh color. Some individuals were, however, dis-

tinctly yellowish throughout their entire length. They were very active

and when disturbed showed vigorous writhing movements involving

strong side to side motions. Serial sections and dissections showed that

the specimens repffesent Fridericia agilis Smith. Aside from certain

variations in size they agree with the original description of the species

in every respect. The original description gives the variation in length of

well-extended living specimens as 25-30 mm. The Kilburn material

showed a range of from 20 to 29 mm., with an average of 24 mm. However,

these measurements were made on alcoholic material and possibly show

a lower range because of a certain amount of contraction during the

killing process. The original description gives the number of somites as

57-66, the average being 62, while the Kilburn material shows a range

of 52-69, with an average of 57. The diameter of the body in the region

of the clitellum is 0.61 mm. -0.75 mm., average 0.68 mm.

This is the first time that F. agilis has been taken since the original

material was collected by Professor Frank Smith (1895) in the vicinity

of Havana, Illinois. .

94 paul s. welch

Enchytraeus albidus Henle

On June 5, 1914, enchytraeids were found in connection with the roots

of house plants at Houghton, Michigan. Professor R. H. Pettit, Michigan

Agricultural College, sent to the writer sexually mature specimens which

proved to be typical Enchytraeus albidus Henle, the first record of its

occurrence in North America west of the Atlantic Coast. Previous North

American records have been discussed by the writer (1917, p. 120-121)

in an earlier paper. The meager data accompanying the specimens give no

information as to their original source. Their occurrence in connection

with the roots of house plants makes it uncertain whether they have

been transported thither with potted plants or are present in the native

soil.

LITERATURE CITED

Delphy, M. J.

1919 Recherches sur les Oligochetes Limicoles. Bull. Mus. d'Hist. nat., No. 7, 7 pp.

1921 Etudes sur I'Organization et le Developpement des Lombriciens Limicoles Thalas-

sophiles. 137 pp. 65 fig. Imprime pour I'Auteur. Paris.

MiCHAELSEN, W.

1900 Oligochaeta. Das Tierreich. 10 Lieferung, I-XXIX, 575 pp. 13 fig. Berlin.

Moore, J. P.

1895 Notes on American Enchytraeidae. I — New Species of Fridericia from the

Vicinity of Philadelphia. Proc. Acad. Nat. Sci. Phil., pp. 341-345. 1 pi.

Smith, F.

1895 Notes on Species of North American Oligochaeta. Bull. 111. State Lab. Nat.

Hist., 4:285-297.

Smith, F. and Welch, P.S.

1913 Some New Illinois Enchytraeidae. Bull. 111. State Lab. Nat. Hist., 9:615-636. 5 pi.

Stephenson, J.

1911 On some Littoral Oligochaeta of the Clyde. Trans. Royal Soc. Edinburgh,

48:31-65. 2 pi.

Welch, P. S.

1917 The Enchytraeidae (Oligochaeta) of the Woods Hole legion. Trans. Am. Micr.

Soc, 36:119-138.

PRIMITIVE MICROSCOPES AND SOME EARLY

OBSERVATIONS^

By

William A. Locy

Northwestern University

The question "Who first constructed the microscope?" is not one of

major importance. The story is somewhat involved. However, the

period in which magnifying glasses were brought into general use for the

study of nature is quite well established. This was near the close of the

sixteenth and in the first part of the seventeenth century. Primitive

microscopes and pioneer observations with these instruments are of

unusual interest, because they represent the tools employed and the

beginnings of a new kind of scientific knowledge. Nothing of this kind

comes down to us from antiquity. We should like to believe that Aristotle,

the Alexandrines, and Galen had means of increasing their natural vision,

but no such evidence exists. The unexpected discovery of so many

appliances of antiquity has placed the modern mind in a receptive condition

to all sorts of suggestions regarding the equipment of the ancients.

A lens-shaped rock crystal, discovered by Layard in the ruins of the

palace at Nineveh, has been heralded as a quartz lens of great antiquity.

This antique ornament or jewel, dating from 721-705 B.C., is now in the

British Museum, and, as Myall, Charles Singer, and others have pointed

out, its surface is not ground smooth but is cut into small facets, which

disperse the light, so that it cannot act as a lens. Moreover, this piece of

quartz is not clear but is clouded by dark bands. "From a number of sites

of classical antiquity crystal balls have been recovered and these may or

may not have been used as burning-glasses. The point is doubtful, but

it is certain that they are not lenses in the usual sense of the word." (Singer.)

The fragmentary and usually dubious references to magnifications by

ancient writers are not satisfying. The most often quoted statement is

from Seneca's Natural Questions (63 A.D.), in which he says: "I may now

add that every object much exceeds its natural size when seen through

water. Letters however small and dim are comparatively large when

seen through a glass globe filled with water." In this connection Seneca

is attempting to explain why the rainbow appears so large, and the rest of

the text shows that he is merely sustaining his hypothesis that objects

seen through water appear enlarged; his mind is not directly concerned with

the magnifying properties of transparent curved objects.

Passing over the story of the use of lenses by Alhazen in the eleventh,

and Roger Bacon in the thirteenth century, we come to the last part of

^ Address of the Retiring Cha!irman of the Section of the History of Science, American

Association for the Advancement of Science, Boston, Dec. 27, 1922.

95

96 WILLIAM A. LOCY

the sixteenth century where we can trace more directly the manufacture

and the use of magnifying lenses. There are various claimants for priority,

but it is not clear to whom the credit belongs. There were a number of

spectacle makers at that time in the Netherlands, Italy, Germany, etc.,

and it would seem that combinations of lenses inserted in the ends of

tubes were happened upon independently by different parties. In these

early days the development of telescopes and of compound microscopes

runs a parallel course. The simple microscope, consisting of a single lens,

appears to have been used before lenses in combination, but both kinds

were often employed by the same observer. After recognizing the English-

man, Digges, (in 1571), and the Hollander, Zacharias (called Jensen),

about 1590, as prominent among the earliest inventors, we venture to

say that to determine who actually was first is a small matter compared

with who first made the instrument the common property of science.

For this honor, perhaps, Galileo has the best claim. He was, says Charles

Singer, the "effective" inventor of the telescope and the compound micro-

scope. About 1608 he made his first telescope (soon followed by enlarged

and improved forms); and with this combination of lenses he not only

made observations on the celestial bodies, but, also, in 1609, published

microscopical observations on minute objects.

We know, as a matter of fact, that single lenses (and lenses in com-

bination) had been used earlier and that the use of magnifying glasses

for scientific purposes came about gradually. A considerable number

of early works exist of insects, spiders, worms, etc., some of them showing

enlargements. For illustration, George Hoefnagel published in 1592 a

set of fifty plates of insects engraved on copper. The pictures had been

exquisitely drawn by his son, Jacob, at the age of seventeen, and some

of them unmistakably indicate the use of magnifying glasses. So far as

known the pictures of Hoefnagel are the earliest printed figures of magnified

objects. There is reason to believe, however, that the naturalist, Mouffet,

had made an earlier use of magnifying lenses. His "Theater of Insects"

("Insectorum sive Animalium Minimorum Theatrum") was prepared

in manuscript as early as 1590 but was not published until 1634. Some

of the illustrations in this book show magnifications.

In the complicated question regarding the invention of microscopes,

involving conflicting accounts, Charles Singer offers some deductions as

follows: 1. The invention of the microscope probably preceded that

of the telescope. 2. The invention of the microscope was the work of

Zacharias Jensen, after 1591 and before 1608. It was perhaps formed of

two convex lenses. 3. This invention was followed by that of the telescope,

about 1608, by Lippershey and Metius. Its military application drew

attention to it. 4. The first telescope was of the Galilean type concave

eye-piece and convex o1)jective. Galileo, however, made both the telescope

PRIMITIVE MICROSCOPES

97

and the microscope the property of science and was the efedive discoverer

of both. His instrument was improved by Kepler in 1611. The priority

of effective demonstration of the telescope rests with Galileo and of the

publication of a mathematical analysis with Kepler.

There is plenty of documentary evidence from writings in English,

French, German, Dutch, and Italian to establish the fact that the use of

the simple microscope was common in the first half of the seventeenth

century. By the time of Harvey evidently magnifying glasses were no

novelty. In his "De Motu Cordis et Sanguinis" (published 1628), he

speaks in a matter of fact way in two places of his use of magnifying

glasses.

A few years later we have the earliest printed pictures of microscopes,

when, in 1637, Descartes published his "Dioptrique" as an appendix to his

well-known "Discourse on Method" and supplied two pictures with

descriptions of microscopes. Fig. 1 shows Descartes' picture of a simple

Fig. 1. Earliest known printed picture of the simple microscope. Descartes, 1637.

(After Petri.)

lens provided with a means of illuminating the object to be examined. H

represents the eye, in front of which, at A, is a plano-convex lens inserted

in a blackened frame; behind the lens is a parabolic mirror with a trans-

parent central area, through which the object can be viewed; the parallel

rays of light from the mirror coming to a focus at the point, E. The

object to be examined is attached to an object-holder, G, at the point

of greatest illumination.

In addition to the foregoing, Descartes published a sketch of a huge

clumsy apparatus designated an "ideal microscope." As shown in Fig. 2,

98

WILLIAM A, LOCY

this has a sliding tube carrying a combination of lenses; the lens near the

eye being plano-concave, and that at the far end of the tube (R) plano-

convex. For illuminating the object, there was a concave mirror, similar

to that of his simple microscope, and also a plano-convex lens placed in

the pathway of light and giving a strong illumination at the point Z.

Descartes says that the single lens may be replaced with one having two

lenses combined. It is evident from these pictures and descriptions of

Descartes that, in 1637, he had represented both the simple and the

compound microscope. The large, unwieldy apparatus was later called

perhaps in derision, a "megaloscope," but so far as known it remained as a

theoretical representation and was never manufactured.

Fig. 2. Descartes' representation of an "ideal microscope," 1637. (Petri.)

The pictures of Hoefnagel and Mouffet, referred to a moment ago,

were merely enlargements of objects visible to the unaided eye, but in the

writings of Athanasius Kircher we have the first authenticated notices

of microscopically minute living organisms. In his "Ars Magna Lucis et

Umbrae," published in 1646, he describes a sphero-hyperbolic lens with

which he made his first observations. Later he used an improved com-

pound apparatus. Speaking of the different kinds of microscopes known

in his time, Kircher says that some use two convex lenses; others use large

glass globes filled with water and still others use a new and clever dis-

covery of the smallest glass globules not larger than the smallest pearl.

With the aid of lenses Kircher saw minute "worms" in all decaying sub-

stances, in milk, and in the blood of [)ersons stricken with fever.

In 1658, in his "Scrutinium Pestis," Kircher gave a notable anticipation

of the germ theory of disease. He described living "corpuscula" as occur-

PRIMITIVE MICROSCOPES

99

ring in great numbers in the blood of plague stricken persons and stated

that these micro-organisms were the source of contagion. Kircher did

not see the organisms that produce bubonic plague — which were discovered

a long time afterward — the structures which he saw were probably pus-cells

and rouleaux of blood corpuscles, but he did ascribe contagion to living

organisms (contagium animatum). More than one hundred years earlier

"with remarkable clairvoyance," Fracastorius had attributed diseases

to mihute bodies or spores but he did not regard them as living organisms.

Kircher's opinion was fortified by his actual observation of minute "vermi-

cula" occurring in all putrifying substances and in the blood of the sick;

his conclusion had some observational basis and his idea that infection is

due to living organisms was a remarkable anticipation which has received

merited attention in recent times. In following this idea of infection

from living organisms, we note that a hundred years later, in 1762, Plenciz

believed that there was a particular organism (seminarium) for each

disease with a definite incubation period, but this noteworth example of

prevision (together with others of similar import) was forgotten and

the matter subject was revived only in the nineteenth century.

We now look with interest on the picture of Kircher's early microscopes.

Fig. 3 from his "Ars Lucis et Umbrae" shows a short tube with a lens

Fig. 3. Kircher's microscope, 1646. (Petri.)

at one end and a plain glass at the other. Another picture, Fig. 4, shows

ornamentation of the tube. The object to be examined was placed against

the flat glass and the lens near the eye was the magnifier. This is the

prototype of the simple microscope. Because they were first used for

100 WILLIAM A. LOCY

magnifying insects, these instruments came to be known as flea-glasses,

and fly-glasses (vitrea pulicaria, vitrea muscaria, etc.). They were small

tubes not thicker and longer than the thumb. In the last part of the

seventeenth century they had quite a vogue as instruments of diversion,

and documentary evidence shows that in 1679 microscopes with spherical

lenses (microscopia globularia) were on sale in Paris.

Fig. 4. An early "flea-glass" with ornamentation of the tube. Zahm, 1685.

In connection with Kircher, we should mention Schott, his colleague

and fellow member of the society of Jesus. Kircher being occupied with

another work besought his friend, Schott, to finish for him and publish a

work on natural magic; this was done, and, in 1657, a year before Kircher's

"Scrutinium Pestis" appeared, Schott published a sort of preliminary

volume designated "Magia Optica" and giving credit to Kircher. The

work was translated and printed in German, in 1671. I have had the

use of this German edition through the courtesy of its owner. Dr. A. B.

Luckhardt, of Chicago.

Fig. 5 is a photograph of the plate of microscopes in Schott's book.

The size of these microscopes has been misconceived on account of the

full-length human figure represented in connection with them and it has

been generally overlooked' that the dimensions of the instruments are

mentioned in the text. Schott says of the picture marked 1 in the cut

that the microscope is a small tube of wood or bone scarcely longer and

thicker than a finger ("das kaum lenger und dicker ist als ein finger Glaich") .

At the end near the eye it is provided with a small spherical glass not

larger than the smallest pearl. The others also are described as relatively

small. The dimensions of picture 4, the largest one represented, is given

as having a tube a foot long and thicker than the thumb mounted perpen-

dicularly on a small block three feet high. These instruments were not

huge "megaloscopes" as represented in Descartes' "ideal miscroscope."

PRIMITIVE MICROSCOPES

101

The presumption is that the artist inserted an entire human figure in

place of the single eye commonly shown in many similar pictures.

In other sources of the nearby period we have an occasional mention

of the size of the instruments employed. For illustration, Hooke's com-

pound microscope (Fig. 11, about 1660), had a tube six or seven inches

long, and a picture supposed to represent the microscope of the Italian,

Divini, shows an instrument provided with five lenses, the length of which,

by different writers, has been estimated from one foot to sixteen and

one-half inches. In connection with the introduction of the microscope

as a tool of science there naturally comes the discovery of micro-organisms,

both animals and plants, and also the minute structure of tissues, of

organic and of mineral substances.

Fig. 5. Microscopes from Schott's Magia Optica, 1657. (Petri.)

The first to devote a long life to studies with the microscope, and to

make a large number of observations — sometimes illustrated with sketches

— was the Dutch observer, Antony van Leeuwenhoek of Delft. Through

his multitudinous observations, published chiefly in the Transactions

of the Royal Society of London and extending over a period of forty years,

he made the microscopical world known to a wide circle. We may cluster

about the name of Leeuwenhoek the story of early microscopical obser-

vations— remembering that there were other men who took part in the

development of this kind of knowledge. In particular, Malpighi, the

Italian, earlier in the field than Leeuwenhoek, extended his observations

to the embryology of animals, to the minute structure of plants, to circu-

lation of the blood in the transparent lungs of the frog (1660), etc., and

Swammerdam, who used lenses extensively in investigating the structure

of insects.

102 WILLIAM A. LOCY

Leeuwenhoek made his observations with small microscopes of his

own contrivance. Although he made several hundred of these instruments

for his own use, he was not, as represented in Dr. Carpenter's article in

the ninth edition of the Encyclopaedia Britannica, an optician, nor a

manufacturer of lenses for the market. Time does not permit now to

demonstrate this point.

Twenty-two years before his death, Leeuwenhoek designated twenty-six

of his microscopes to go to the Royal Society after his death. His com-

munication to the Royal Society was dated Aug. 2, 1701, and since it

throws light on the extent to which he prepared his own instruments,

it is worth quoting: "I have (says Leeuwenhoek) a small black cabinet,

lacker'd and gilded, which has five little drawers in it, wherein are contained

thirteen long and square tin boxes, covered with black leather. In each

of these boxes are two ground microscopes, in all six and twenty; which

I did grind myself, and set in silver; and most of the silver was what I had

extracted from minerals, and separated from the gold that was mixed with

it; and an account of each glass goes along with them.

"This cabinet, with the aforesaid microscopes, (which I shall make

use of as long as I live), I have directed my only daughter to send to your

Honors, as soon as I am dead, as a mark of my gratitude, and acknowledg-

ment of the great honor which I have received from the Royal Society."

Baker, in his work 'The Microscope Made Easy' (1742), mentions

having had these instruments away from the rooms of the Society for ex-

amination. He described them and figured some of them, but soon after

they were lost sight of, and, unfortunately, these hierlooms to science

have never been recovered.

Inasmuch as Baker had these microscopes under observation his

testimony as to the shape of the lenses is important. He says: "Several

writers represent the glasses Mr. Leeuwenhoek made use of in his Micro-

scopes to be little globules, or spheres of glass; which mistake most probably

arises from their undertaking to describe what they had never seen; for,

at the time I am writing this, the cabinet of Microscopes left by that

famous man, at his death, to the Royal Society as a Legacy is standing

upon my table; and I can assure the world that every one of the twenty-six

Microscopes, contained therein, is a double convex lens, and not a sphere

or globule."

Leeuwenhoek gave descriptions and some drawings of his microscopes,

and those in existence have been described and figured by different writers,

so that we have a very good idea of his working equipment. He preferred

the single lens, with a small glass of marked curvature, giving a small

field but clearer definition than the compound microscope of Hooke. He

made different microscopes to suit his purposes, having a range of magnifi-

cation from 40 to 270 diameters.

PRIMITIVE MICROSCOPES

103

One of Leeuwenhoek's originals exists at the University of Utrecht,

and at my request Professor H. F. Nierstrasz photographed this instru-

ment natural size. Three views of his photographs are shown in Fig. 6.

Fig. 6. The Leeuwenhoek microscope in the University of Utrecht.

Professor Nierstrasz.

Photographed by

The instrument has two small copper plates, perforated by an orifice in

which the small, nearly spherical lens is inserted. In the original, the

copper plates measure one inch broad and a little short of two inches

long. The object-holder is represented in the lower right-hand figure as

thrown to one side. By a vertical screw the object could be elevated or

lowered, and by a transverse screw it could be brought near or removed

farther from the lens and thus be brought into focus.

In use, the instrument was held close before the eye (Fig. 7) against

the light, and the object was viewed by transmitted light.

Fig. 7. To show how the Leeuwenhoek microscope was held. (Petri.)

104

WILLIAM A. LOCY

In some instances, however, the microscope was provided with a

concave reflector (Fig. 8) similar to that used by Descartes, to illuminate

the object by reflected light.

Fig. 8. A Leeuwenhoek microscope provided with a concave reflector. (Petri.)

Fig. 9 shows the way in which the microscope was arranged by Leeu-

wenhoek to examine the circulation of blood in the transparent tail of a

small fish or tadpole. The animal was placed in water in a slender glass

Fig. 9. Leeuwenhoek 's arrangement for e.xamining circulation of the blood.

tube, and the latter was held in a metallic frame to which a plate (marked

D) was joined, carrying the magnifying glass. The latter is indicated in

the circle above the letter D, near the tail-fin of the animal. The eye of

the observer was applied close to the lens which was brought into position

and adjusted by means of screws.

Of the many discoveries of Leeuwenhoek, we can give only one example.

This will be his observation of the bacteria; since it is the earliest account

of bacteria accompanied with sketches, it is of especial interest. The

PRIMITIVE MICROSCOPES

105

discovery of these minute forms was a feat of trained observation, and it

is remarkable that Leeuwenhoek, with his primitive equipment, was able

to see them and to describe them so clearly. One of his letters of 1681

indicates that he had seen bacteria at that date, but his formal description

of them came in 1683. There can be no doubt from his sketches and

descriptions that he saw the chief forms of bacteria — round, rod-shaped and

spiral forms.

His first observations on bacteria were communicated to The Royal

Society of London, in a letter dated Sept. 17 (not 14), 1683, and published

in the Philosophical Transactions for the year, 1684.

A photograph of the cut published with his observations is shown

in Fig. 10. It may be remarked in passing that the reproduction of the

cut by Loffler, Petri, and others, is not quite facsimile and their quotations

A ^

J)

-Jt.

'6f-3-

M

'"S:J'"JiL

Fig. 10. Photograph of the original plate of bacteria as seen by Leeuwenhoek in 1683.

(After Charles Singer, from the Philosophical Transactions, 1684.)

do not correspond verbally with the text in the Philosophical Transactions.

A few lines from the original publication in the Philosophical Transactions

shows the objective quality of Leeuwenhoek's descriptions:

"Tho my teeth are kept usually very clean, nevertheless when I

view them with a Magnifying Glass, I find growing between them a little

white matter as thick as wetted flour: in this substance tho I could

not perceive any motion, I judged there might probably be living creatures.

"l therefore took some of this flour and mixt it either with pure rain

water wherein were no animals; or else with some of my Spittle (having

no Air bubbles to cause a motion in it) and then to my great surprize

perceived that the aforesaid matter contained very many small living

106 WILLIAM A. LOCY

Animals, which moved themselves very extravagantly. The biggest sort

had the shape of A (see the cut). Their motion was strong and nimble,

and they darted themselves thro the water or spittle, as a Jack or Pike

does thro the water. These were generally not many in number. The

2d. sort had the shape of B. These spun about like a top, and took a

course sometimes on one side, as shown at C and D. They were more in

number than the first. In the 3d. sort I could not well distinguish the

Figure, for sometimes it seem'd to be an Oval, and other times a Circle.

These were so small that they seem'd no bigger than E. and therewithal

so swift, that I can compare them to nothing better than a swarm of

Flies or Gnats, flying and turning among one another in a small space.

Of this sort I believe there might be many thousands in a quantity of

water no bigger than a sand tho the flower were but the 9th. part of the

water or spittle containing them."

"Besides these Animals there were a great quantity of streaks or threads

of different lengths, but like thickness, lying confusedly together, some

bent, and some straight as at F. These had no motion or life in them, for

I well observed them, having formerly seen live-Animals in water of the

same figure."

Leeuwenhoek extended his observations to others: two women; a

child of 8 years old; the spittle of "an old man that had lived soberly;

and another old man who was a good fellow." The "meal" between

the teeth of the old men "had a great many living Creatures, swimming

nimbler than I had hitherto seen. The biggest sort were numerous, and

as they moved, bent themselves like G. The other sorts of Animals were

in great numbers insomuch that tho the meal were little, yet the water

that it was mixt with seem'd to be all alive, there were also the long

threads above mentioned."

The figure marked "H" has very generally perplexed writers, and has

been designated by some as a representation of those round bacteria which

occur in packets of cubes (sarcinae), but later in the same paper, Leeuwen-

hoek says that "H" represents scales of the outer skin (cuticula).

It is worthy of note that bacteria were pictured before protozoa (which

had been discovered by Leeuwenhoek in 1675), and if we except the poor

picture of a shelled-protozoan, (Rotalia), by Robert Hooke, in 1665,

they were, I believe, the first micro-organisms to be illustrated in printed

pictures.

Fig. 11 is a picture of Robert Hooke's compound microscope, made

about 1660, and constituting the frontispiece of his Micrographia, which,

in 1665, was published as the first book devoted expressly to microscopical

observations. This shows the form to which the compound microscope had

attained in the last part of the seventeenth century. Space does not

permit us to follow its further development through the eighteenth and

PRIMITIVE MICROSCOPES

107

nineteenth centuries. Hooke's Micrographia gave a real impetus to obser-

vations with the microscope, especially in England. Among others, Neimiah

Grew, the fellow countryman of Hooke, was stimulated by its publication

to carry on his extensive observations on the microscopic structure of

plants.

The psychological influence of the use of the microscope was very

great. By sharpening attention and directing it towards definite points,

the powers of mental application were improved and impressions received

through the sense of sight were made more exact. Now, perception through

U-

Fig. 11. Hooke's compound microscope (about 1660). From his Micrographia, after

Carpenter.

trained senses is the foundation of all scientific knowledge, and, as a

matter of fact, we find the early workers with the microscope, Robert

Hooke, Malpighi, Grew, and Leeuwenhoek, seeing nature more scientifi-

cally and exactly than their predecessors. As Sachs remarks in his History

of Botany: "Preception by the use of the optic nerve had to be accom-

panied by conscious and intensive reflection, in order to make the object,

which is observed only in part by the magnifying glass, clear to the mental

eye in all the relation of the parts to one another and to the whole. Thus

the eye armed with the microscope became itself a scientific instrument,

which no longer hurried lightly over the object, but was subjected to

severe discipline by the mind of the observer and kept to methodical work."

Although there was started a period of more incisive observation, the

early microscopes were very imperfect and it was not until their improve-

ment in the first third of the nineteenth century that the full effect of

their use was realized.

AN ILLUMINATING DEVICE FOR MICROSCOPES

By

William A. Beck

University of Dayton

Introduction

Much attention has been given by the builders of microscopes to the

selection of the correct combination of objectives and oculars for the

study of various preparations. The importance of proper illumination has

also been appreciated and much thought and care has been expended

upon this phase of microscopic methods. There are difficulties presented,

however, because of the very nature of things, so that much remains to be

desired in modes of illumination for certain fields of investigation. The

device which I am bringing before the profession is an effort to improve

the illumination.

Modes of Illumination

Bright Field Illumination.

About 95% of the study of microscopic objects is done in a bright field.

The entire field is lighted and the objects to be studied appear as colored

objects, with the different tones severely bounded, each for itself being

homogeneous within its own bounds. In extreme cases the objects stand

out as silhouettes in their bright field. The very nature of the microscope

has predetermined the development of this mode of illumination because

of the relatively short working distance of the objectiv^es. Any other mode

was too limited in its application, particularly during times when an

intense and concentrated source of artificial illumination was impossible.

The bright field illumination is obtained by allowing the light from the

sun or some artificial source to fall directly or after diffusion, upon the

mirror of the substage, whence it is directed, with or without the aid of a

condenser, upon the subject under study, which must naturally be trans-

parent or translucent. The light, thus directed, may be transmitted

axially or obliquely or may be so thoroughly diffused by secondary reflecr

tion, that the illumination no longer has the character of being directed.

Dark Field Illumination.

In contrast with this is Dark Field Microscopy. Ordinarily the field is

actually very dark and the object is bright or, in some cases at least, the

light is deflected into the objective and gives the impression of a white

object on a dark background. In other cases the background is not ac-

tually dark or of even tone, in fact it may be brighter than some tones of

the image, but since this is due to secondary reflection under studied con-

108

AN ILLUMINATING DEVICE FOR MICROSCOPES 109

ditions, so as to bring the subject into better relief by the same methods,

we might well consider this mode under the heading of Dark Field Micro-

scopy.

There are two principal cases. First when the objects are self-luminous

i.e., phosphorescent, or when, by illumination with ultra-violet light, they

become phosphorescent. Secondly, when the objects themselves do not

emit light but which reflect or deflect the light, reaching them from some

outside source, passes into the microscope. Different opaque objects

having different reflecting powers, will under these conditions, produce

widely different tones in the images produced. We might do well to borrow

a term from Astronomy in describing them, where albedo is taken to mean

the ratio of the quantity of light reflected to the quantity of light received.

If one looks into the sky and notes the stars against the dark vault, he

has a good illustration of the first case, i. e., of darkground illumination. If he

regards the planets he has an illustration of the second case.

My device pertains chiefly to the domain of Dark Field Microscopy and

in particular to the Second Case.

History of Illumination of Opaque Objects

For progress in any field, the first step must always be the investigation

of the principles involved. Later we find that development of knowledge

in other fields are helpful in making proper application of these principles,

in the attainment of a given end. The proper illumination of opaque ob-

jects under microscopic examination is no exception.

Early investigators appreciated that the principle of contrast was of

the highest importance in rendering objects visible. The application of

convex lenses for magnifying the objects was made by Roger Bacon as

early as 1266. In 1610 Kepler devised the compound microscope with

convex objective and convex ocular from which, in time, evolved the

modern form. In this evolution, condensers, for lighting the objects,

played a prominent part. In 1637 Descartes used a large parabolic mirror

to direct sunlight upon the object. This mode of lighting showed the ob-

ject more or less bright, on a dark background. His unwieldy apparatus

gave way to more convenient microscopes and condensers, as progress was

made in the various domains of science. It is interesting to note, however,

how men have struggled down to the present time for a better application

of a principle, recognized at an early period.

Lister appreciated the fundamental difference between Bright Field

Microscopy and Dark Field Microscopy in 1830. It was not until Zeiss

in 1904 and Leitz in 1905 made practical applications, which were later

discarded for more effective devices, so that the object is illuminated, by

beams of light, in such a direction with reference to the axis of the objec-

tive, that none of them can enter the objective directly and the light, going

110 WILLIAM A. BECK

into the microscope, comes only from the objects themselves, so that they

appear self-luminous on a dark ground.

When the light is directed from below upon the object we do not

expect to obtain a clear image of the upper surface of the object but are

more concerned about discovering the existence of such objects and note

their movements etc. The application of this principle has led to the

wonderful developments in Ultra Microscopy.

If the light is directed upon the object from above and the object is

over a non-reflecting background, the object will appear bright in a dark

field.

From the time of Descartes, many efforts have been made to improve

the mode of illumination from above. Reflectors were used to direct the

light from the side onto the object, or again a "Bull's Eye" Condenser

concentrated the light, from a specific direction, onto the object.

The Lieberkuhn reflector which was devised in 1740 is not unlike Des-

cartes' device, in principle, where a parabolic mirror directs the light

onto the object. Since 1850 two additional devices for illuminating from

above, have come into use.

In 1852 Riddell suggested introducing the light from the side into the

objective and reflecting it down upon the object. From this suggestion

have resulted the various types of Vertical Illuminators. Some employ

total reflection in a prism, to direct the light down through the objective,

others a disk of glass and still others a mirror. A short time ago a new

method was devised by Professor Alexander Silverman of the University of

Pittsburgh. It consists of a circular electric filament lamp, Which surrounds

the objective and shines down upon the object.

The Illumination of an Object Under Study

Up to the present only directed illumination was employed in the mi-

croscopic study of opaque objects, to obtain a knowledge of their surface

configuration.

The directed light was either oblique to the optic axis of the system of

lenses or approximately parallel, i.e., either oblique or vertical with respect

to the surface under examination. We might then simply use the terms

oblique or vertical, to designate these modes of illumination. Both are

to be considered as directed illumination, because we have a pencil of

light from a definite direction.

Oblique Illumination.

Oblique illumination can be obtained either by means of a reflector

attached to the objective or by directing rays from a radiant, lying above

the plane of the surface of the object. When a radiant is employed, for

example an arc lamp, a tungsarc, or a filament lamp, a condensing lens is

AN ILLUMINATING DEVICE FOR MICROSCOPES

111

usually interposed, between the light and the object, in order to concen-

trate the light rays and to facilitate the proper placing of the beam. If we

are dealing with a highly polished surface this mode of illumination has no

value, because no rays can enter the objective according to the laws of

regular reflection (Figure 1).

Fig. 1

Illustrating the law of regular reflection and showing how oblique illumination on a flat

surface cannot direct the light into the microscope.

Fig. 2

Illustrating regular reflection on an irregular surface, showing how regions at the proper

angle reflect an intense light into the tube of the microscope while others do not.

points in close pro.ximity may produce defraction patterns.

Many

If the surface of the object illuminated is irregular or etched, the rays

entering the objective from some points are very intense, while from

others they may be entirely lacking and therefore do not express the

relative albedo of the surface, nor give an impression of the plasticity of the

object. This becomes clear at once from a study of figure 2. This figure

also shows how certain regular markings in certain cases produce diffrac-

tion effects. Diffraction patterns make it difficult to interpret the true

structure.

The greater the obliquity the greater the incident difficulties become,

so that with high power objectives, where the free working distance is

very small, this mode of illumination is entirely out of the question.

Illuminating from several sides tends to correct somewhat the dif-

ficulties referred to, in giving the body illuminated more of the character

of a self luminous body, according to Huygens' impression, that each point

illuminated becomes a new point source of disturbance. Then again dif-

112

WILLIAM A. BECK

fraction patterns are eliminated. Complete annular illumination was

attempted in the paraboliod illuminator which was very popular at one

time. For correct illumination, the curvature of a reflector placed around

the objective would have to vary for each working distance. These were

later almost entirely superseded by the vertical illuminators.

In the Silverman device, that has come into the field in relatively

recent years, and which became popular immediately upon its appearance,

the illuminant is attached directly to the objective. Here the illumination

is about three fourths annular and consequently shows decided improve-

ment over previous modes of illumination by the oblique method. There

still remain these difficulties: the illumination is not completely annular.

The angle of illumination cannot be varied at will. The intensity of il-

lumination is limited. The light cannot be completely diffused. The light

cannot be filtered at will. The life of the lamp is short. A considerable

amount of undesirable heat is developed.

Fig. 3

Showing a rough surface under vertical illumination. Note that only a strictly horizontal

surface can return the light directly into the tube of the microscope. Diffraction patterns

cannot produce images in the field.

Vertical illumination is obtained by placing a reflecting surface in a

mounted cell attached to the microscope, just above the objective. The

reflector sends the illuminating beam of light through the objective, which

acts as a condenser, concentrating the light rays into a bright spot of light,

upon the surface of the object, at a point lying, appro.ximately, in the

optic axis of the microscope. From the surface of the object, the rays

are reflected back through the objective and form the image of the object

in the usual manner.

If the object to be studied is absolutely flat, it is evident that the

rays reflected from the object might be considered as emitted by the

object and these should form an image, that should give a correct impres-

sion of the albedo of the various parts of the field. Furthermore, if there

AN ILLUMINATING DEVICE FOR MICROSCOPES 113

are sections, that otherwise would be inclined to form diffraction patterns,

the law of regular reflection would require that all reflected and diffracted

rays fail to enter the objective, not being parallel to the optic axis. (Figure

1, central beam. See also Figure 3 where points lie in close proximity.)

There is absolutely no question that this type of illuminator has rendered,

and is still rendering, immeasurable service to science. For certain work

it may never be replaced.

When the object to be studied is not flat, the image immediately ceases

to give the correct impression regarding albedo and the nature of the

surface. The image needs interpretation. The investigator may be dealing

with more or less highly polished surfaces and with areas, part of which,

are polished, part rough and often studded with minute points. Some-

times cracks or cleavage planes may cross the field.

With ordinary etched surfaces, polished portions appear bright and

etched surfaces less bright. The glare from the brighter surfaces, seeks to

lessen the definition in the less bright fields. It is questionable if the

detail of the mat surfaces are in any degree accurate. To demonstrate

fissures, cleavage planes, depressions etc., it becomes necessary that the

examination with the vertical illuminator be supplemented by oblique

illumination and to study the direction of the shadows with respect to the

radiant, remembering of course that in the image seen in the microscope,

directions are completely reversed. The need of this study suggests a mode

of oblique illumination, such that the light beam can be turned about the

axis of the microscope, with least possible difficulty.

The Illumination of Macroscopic Objects

Since the principles for the correct illumination of objects, as we ob-

serve them in every day life, with the unaided eve, are the same as for

microscopic study, we might do well to give a little attention to the phe-

nomenalprogress that has been made within the past decade by illuminating

engineers in their proper sphere.

Prominent among the scientists who are working for better illumination

to obtain correct values and at the same time to save the eyes, is Professor

C. E. Ferree of Philadelphia. He has made a series of tests of the eye,

under different kinds of illumination, daylight, indirect lighting, semi-

direct lighting and exposed filament lighting.

He found that after three hours' work under day light, the eye lost

practically nothing in seeing efficiency. Under indirect lighting the

effect was almost the same; the eye was 91% efficient in seeing ability.

Under exposed direct or semi-indirect lighting the loss in seeing efficiency

was enormous; the remaining efficiency being only 25% with semi-direct

and 14%, with direct. Science has conclusively demonstrated, that if we

would preserve our eyesight and obtain correct values of the tones in a

114 WILLIAM A. BECK

scene we must avoid, so far as possible, glare and direct illumination, be-

cause the eye cannot endure excessive light. Momentary blindness results

from intense light because the entire retina or a portion of it becomes

paralysed. The result is that we do not perceive objects within the range of

vision that should be seen normally. The greater the glare of any spot in

the field, the more intense the illumination must be before the other

portions can be perceived, which increase however, serves to render the

"spot" more glaring, thus defeating the purpose.

The Formation of an Image.

If in the neighborhood of a luminous point P, there are refracting and

reflecting bodies having an arbitrary arrangement, then, in general, there

passes through any point in space, one and only one ray of light, i.e., the

direction which it takes from P to P' is completely determined. Under

given conditions, certain points may be formed, at which, two or more of

the rays emitted by P intersect in a point P' which is called the optical

image of P. If we have many points in our object under study, the summa-

tion of all the images of these points will give a more or less accurate replica

of the object, according as the numbers expressing rays, passing through

the P's do or do not bear the same ratios to each other, as the numbers

expressing the rays actually emitted by the conjugate points in the object.

From what has been said of regular reflection, it is clear that we cannot

form an adequate im.age of the reflecting object when it reflects regularly,

but only of some other object that emits rays of light which come to us

after reflection.

When the light falls in a definite direction on an unpolished surface it is

reflected in various directions. The amount of light going in a given direc-

tion for the formation of an image depends upon the position of the image

with respect to the position of the illuminant. It is clear then that the

number of rays that combine to form an image, depends upon the particu-

lar position of the images, as well as upon the nature of the surface and the

albedo of the object. This varying factor, of position, offers serious

difficulty in the formation of a correct image. The thought naturally

arises in the mind, that we attempt to illuminate equally from all sides in

order that only the nature of the surface and the albedo shall determine

the image.

Reflection of diffused light, produces not only correct form and tone,

but is also selective, so that we maintain correct color value. The rich red

petal of the geranium thus illuminated by white light, reflects diffusely in

all directions only the red rays. In order to form correct images it follows

from the above discussion that direct illumination must be avoided and

diffused illumination employed, whenever possible.

AN ILLUMINATING DEVICE FOR MICROSCOPES

115

Illuminating engineers corrugate reflectors and give the walls a rough

finish rather than an even finish to avoid regular reflection. All their

efforts are directed in distributing the light energy, like a fine spray,

throughout the entire room. We are all familiar with the excellent results

obtained by the modern modes of illumination.

Fig. 4

A study under Northern Sky illumination. Note the exactness of detail and the plastic-

ity. The snow helps to diffuse and illuminate, so that the illumination is practically annular

and diffuse. Copyright by John Mathews.

Photographers have long ago known that they obtain their best pic-

ture by the evenly distributed illumination from a northern sky Figure 4.

The commercial photographers recognize the importance of completely

annular illumination and diffuse illumination and employ methods in

which they obtain so called "cross-fire" of light, by which term they mean,

that the light comes from all directions with almost equal intensity.

116

WILLIAM A. BECK

To illustrate the differences between obliquely directed illumination

and vertical illumination and "cross-fire" illumination a bust of Schiller,

which was painted with a dull even white, was photographed under the

three conditions. Just as was expected marked differences were noted.

The picture that was taken with the aid of oblique illumination, was so

badly distorted by extreme contrasts and heavy shadows that it gave no

impression whatever of Schiller's physiognomy. The vertical illumination

produced a picture that was more even and lacked that extreme contrast,

in fact was flat. The greatest difticulty with the picture was that it gave a

false impression of the real appearance ot the subject, because elevations

Fig. 5

This diagram illustrates how the light, from vertical iUumination upon an elevation, fails

to return to the observer or the instrument.

were dark and depressions were bright. This is just as it should be. A hill

would cause the light to be dispersed while the valley would return too

much of the light. This is clearly shown in figures 5 and 6. The third

picture was of the nature of those ordinarily produced by portrait photog-

raphers, marked by correctness of form and surface variations and a

remarkable plasticity.

The chief aim in view in designing the new type of illuminator was to

obtain a diffuse or "cross-fire" illumination for microscopic objects, of a

sufficient intensity to obtain in pictures, of the same relative value as in

the third case of the photography of the bust.

The Illuminating Device

The device consists of a glass medium which completely surrounds the

object under study (Figure 7). The light enters at the ground surface A,

is totally reflected at the surface C, and is finally refracted at the surface

D, and thus directed from all sides upon the object under study. The

object rests upon a stage Ei, which is adjustable, so as to allow a variation

AN ILLUMINATING DEVICE TOR MICROSCOPES

117

in illumination. The illuminator can be made of any convenient size. The

one which I actually employ and which I recommend for most work, is

so designed as to fit the sleeve on the substage, which ordinarily receives

the Abbe condenser.

Any convenient light source may be employed. For ordinary study, a

common filament lamp or the microscope lamp is satisfactory. With the

gas-filled lamp, one can obtain greater intensity. Just as our modern

illuminating engineers seek a great quantity of light very finely distributed,

so also it is very advantageous to use an arc lamp in connection with the

illuminator, distributing the intense light very evenly. The illuminator

allows the microscopist to employ practically any intensity of light he

desires.

Fig. 6

This diagram illustrates how the valley returns too much of the light under vertical

illumination and causes the values of the field to be reversed.

Plane parallel light is ordinarily to be recommended, which may be

obtained by placing a lens in such a position that the light source is in the

principal focus of the lens. By varying the position of the lens, one can

readily vary the nature of the illumination and in a very interesting man-

ner, simulate "spot-lighting." Direct or diffused sun-light can be em-

ployed as well as an artificial light source.

The light may be allowed to fall immediately upon the surface Ai or

to be reflected, from the mirror of the substage, onto that surface. If the

mirror is employed one can "spot out" certain regions of the subject from

one side or the other by simply turning the mirror. This makes it very

easy to study the nature of a given object by means of shadows.

The angle which the side Di makes with the vertical, determines the

deviation of the beam of light from the horizontal as it passes from Ci to

Di- The deviation rnust be such that the entire field to be studied by the

118

WILLIAM A. BECK

a E,

Fig. 7

Longitudinal section of the illuminating device and the plan of the same. Bi is the

glass medium mounted in a metal sleeve. The light enters the medium at \\ and is totally

reflected at the surface Ci and thus directed to the surface Di. From Di the light deviates

from the horizontal direction and is directed to the stage Ei. The entire piece is inter-

changeable with the .'\bbt; condenser.

AN ILLUMINATING DEVICE FOR MICROSCOPES

119

determined objective, be sufficiently illuminated after the objective is in

the position of the correct working distance.

The dispersion of the light depends upon the refractive angle of the

device which is given by the angle which the face Di makes with the verti-

FiG. 8

Another form of the illuminator. This one is made to fit the large opening on the stage

of the microscope when the entire mechanical stage is removed. Note the three different

refracting angles where the light leaves the glass medium. The cone Ai converts the solid

cyhnder of light into a hollow cylinder of light. This form can be made in one or more pieces.

The light is received at Ao, transmitted at Bj, reflected at C2, again reflected at D2 and

refracted at the surfaces Ej.Fj and G2, falling at different angles, upon the subject resting on

H2.

cal. This angle is always small and the angular dispersion in consequence

is negligible, so that we have a condition of achromatism without special

correction.

120 WILLIAM A. BECK

After a careful study of these values was made for the various objec-

tives, it was found possible to have three refractive angles which would

give highest efficiency for the various objectives that can be employed. If it

is possible for the makers of objectives, to narrow down somewhat the

diameter of the lower portion of high power objectives, there is no reason

why this modification, should not make it possible to study any opaque

object at the highest diameter of magnification, by this method of illumi-

nation.

To increase still further the intensity of illumination, a special design

for the lower portion of the device was made to convert the solid cylinder

of light, into a hollow cylinder of light, by means of a reflecting cone and

totally reflecting surface.

These modifications gave rise to the form shown in figure 8. It is not

necessary that the transparent medium be of one piece as the diagram

indicates. Furthermore the arrangement for a variable inner stage is not

shown. Finally the new device can be arranged in two units. One portion

resembling the illuminator shown in figure 7, and the other representing the

lower portion in figure 8, by means of which the solid cylinder of light is

converted into a hollow cylinder of light.

The light can be diffused by placing a ground glass between the source

and the illuminating device or by placing a diffusion medium around the

object which receives the light from the surface Di, (figure 7).

For certain work it is desirable to filter the light. Regular filters can ,

be interposed between the source of light and the illuminator.

It is to be noted particularly that by this method we obtain completely

annular illumination and that we can without difficulty obtain diffuse

illumination resembling the illumination which photographers seek from the

northern sky or such as is obtained by the devices employed by com-

mercial photographers. If light shadows are desired one can very easily

increase the illumination in a specific direction.

The device can be used simultaneously with the vertical illuminator.

In an interesting experiment, both were set into position and first the

light from the one and then light from the other was employed. It was

interesting to note a complete reversal of the tones as explained above

and indicated in figures 5 and 6.

The device was originally designed for the study of opaque objects but

it can be used to advantage for the study of transparent objects by placing

an uneven reflector, as a piece of filter paper, on the stage E and place the

preparation, in the ordinary way, upon the regular stage of the microscope.

The illumination thus obtained is remarkable for its softness, clearness and

definition. One can study with less eye-strain when employing this method

and the contrast is sharp, even though the light is not intense. This is just

as it should be, according to the principles outlined above.

It is to be hoped that this method of illumination will be helpful in

various fields of microscopic investigations and that the method itself here

outlined in principle, will be further developed.

AX ILLUMIXATIXG DEVICE FOR MICROSCOPES

121

Explanation of Plate

Fig. 1. Brassica juncea "wild mustard."

Fig. 2. Linen cloth. Illumination evenly diffused.

Fig. 3. Scales on wing of butterfly.

Fig. 4. Potassium iodide isometric crystals showing accretion and cleavage planes.

Fig. 5. Bottle cork. Sunlight was used. Illumination evenly diffused.

Fig. 6. Shagreen of Squalus acanthias showing placoid scales. Evenly diffused illumina-

tion.

ABNORMAL SPECIMENS OF HELODRILUS CALIGINOSUS

TRAPEZOIDES (DUCES) AND HELODRILUS

ROSEUS (SAVIGNY)i

By

Bess R. Green

University of Illinois

Introduction

Comparatively few abnormal earthworms have been described, al-

though variations in both external and internal structures are of common

occurrence among the various species of Lumbricidae. The importance

of the study of abnormal development in earthworms lies in the application

of the knowledge thus obtained to the relations that exist in normal develop-

ment. In normal development growth proceeds in such a way as to bring

about the formation of similar structures in the right and left halves of

the same somite. Occasionally the normal method of procedure is inter-

fered with and an asymmetrical arrangement of the organs results. The

organs develop from paired germ-bands which arise practically inde-

pendently on the right and left sides of the body thus making asymmetrical

relations between the organs of the two sides possible. In abnormal worms

the organs may be present in unequal numbers on the two sides, or the

members of a pair of organs many develop in different somites on the

right and left sides. In many specimens there is also a lack of certain

correlations between external and internal structures that occur in normal

individuals.

Variations other than those involving asymmetrical relations are of

two types. Abnormal specimens in which organs vary but slightly either

forward or backward from their normal positions constitute one group or

type. The second group is composed of worms in which the organs appear

in other than normal positions due to a doubling of somites in the anterior

part of the body. There may also be a lack of symmetry between the

organs of the right and left sides in either group. In a recent paper Pro-

fessor F. Smith (1922) described abnormal specimens of H. subrubicundus

and H. tenuis which illustrate the first type of variation, and this paper

treats of two examples of the second type found in H. caliginosus trape-

zoides and //. roseus.

A detailed discussion of the literature dealing with abnormalities in

earthworms will be undertaken later in a more extensive paper. At

present there is not sufficient data to make such a discussion of any great

value.

' Contribution from the Zoological Laboratory of the University of Illinois, No. 219.

122

SPECIMENS OF HELODRILUS CALIGINOSUS 123

The specimens here described are from Professor F. Smith's collection

of abnormal earthworms. These worms were found in the banks of a stream

at Urbana, Illinois.

HeLODRILUS CALIGINOSUS TRAPEZOIDES (DUGES)

The description of this specimen is based on the study of sagittal

sections of the first thirty-three somites. The worm resembles the normal

H. caliginosus trapezoides in the general appearance of the somites and

clitellum, in the relative positions of the setae, and in the presence of

paired, ventral, glandular pads on three of the anterior somites.

External Characters. — This worm which appears to have been injured

at the posterior end measures 11.1 cm. in length and there are 130 somites

present. There is no evidence of somites that are doubled on one side and

not on the other. The setal arrangement in somites 24 to 30 is indicated

by the formula aa:ab:bc:cd = 11:1.5:7.2:1. This is about the usual arrange-

ment for the setae in the species. The saddle-shaped clitellum extends

over somites 32-37 and is very slightly developed on 31 and 38. In a normal

worm it commences on 27, five somites farther forward, and extends

posteriorly over eight or nine somites. The tubercula pubertatis of the

right side is continuous over three somites, 34-36, and the other one

involves three and one third somites, 2/3 33-36. In a normal individual

they are present on somites 31-33. They have the usual relation to the

clitellum since they end posteriorly on the somite immediately in front

of the one on which the clitellum ends. The glandular papillae which

usually include the ventral setae of somites 9, 10, and 11 are found in

this specimen on 10, 11, and 12 on the right side and on 11, 12, and 13 on

the left. The setae included by these papillae are modified and are about

twice as long as ordinary setae and are much more slender. The first

dorsal pore is present on 14/15 which is five somites posteriad of its usual

position. A spermiducal pore surrounded by a glandular papilla is present

on 18 on the right side. A similar papilla is present on each of somites

20 and 21 on the left side and surrounds what appears to be a spermiducal

pore on each of these somites. Normally there is a pair of pores on somite

15. An oviducal pore is present on 17 on the right side, and there is one

on 19 on the left. These positions are respectively three and five somites

posteriad of the normal positions of the pores. On the right side, sperma-

thecal pores are present on 11/12 and 12/13, two somites posteriad of

their usual positions. The spermathecal pores of the left side are on

14/15, 15/16, and 16/17. The first two pores are situated five somites

posteriad of their usual positions, 9/10 and 10/11.

Internal Characters. — Most of the reproductive organs of the right side

of the worm are present in normal numbers but vary from their usual

positions (Figs. 2 and 3). Spermaries are present in 12 and 13; sperm

124 BESS R. GREEN

sacs bearing the usual relations to these organs are in 11, 12, 13, and 14;

and spermathecae are included within the septa 11/12 and 12/13. These

organs are situated two somites posteriad of their respective positions in

a normal worm. The spermiducal pore is on somite 18, which is three

somites posteriad of its normal position. Likewise the ovary, in 16, and

the oviducal pore, on 17, are present three somites posteriad of their

normal positions. The separation of the most posterior spermary and the

ovary by two somites indicates a probable doubling of what would be

somite 12 in a normal worm. Further evidence of the doubling of this

somite is shown by the presence of two vessels uniting the lateral longi-

tudinal vessel with the dorsal vessel, one in 14 and one in 15; while in

normal worms there is but one such vessel, in 12. A pouch of the calciferous

gland, normally in 10, is present in 12 on the right side of this specimen.

It bears the usual relation to the first spermary being included in the

same somite with it.

The reproductive organs on the left side are found five somites pos-

teriad of their respective, normal positions. Supernumerary organs in

excess of the normal number will be mentioned in connection with the

accountof certain reproductive organs and of the "hearts." The spermaries

and spermiducal funnels, usually in 10 and 11, are present in 15, 16, and

an additional one of each in 17. All three of these spermaries are equally

developed. The sperm duct could not be traced to the exterior. Sperm

sacs are present in the posterior part of 14, 15, and 16, communicating

with somites next posterior; and in the anterior part of 16, 17, and 18,

communicating with somites next anterior. An ovary and an oviducal

funnel are present in 18. The oviduct extends from the oviducal funnel

to the oviducal pore on 19. The usual position of the ovary and oviducal

funnel is in 13 and that of the oviducal pore on 14. The spermathecae,

three in number, are included within septa 14/15, 15/16, and 16/17.

Normally there are but two spermathecae present, within septa 9/10 and

10/11. The pouch of the calciferous gland is very slightly perceptible,

if present at all, in 14. The position of the pouch is not in conformity

with its usual relation to the most anterior spermary. Ordinarily these

two structures are present in the same somite.

There are seven "hearts" on the right side and nine on the left. The

most posterior "hearts," which are usually in somite 11, are in 17 on the

left and in 13 on the right side. As above mentioned two vessels arise

from the dorsal vessel in somites 14 and 15 on the right side and join to

form a single lateral longitudinal vessel. A lateral longitudinal vessel was

not seen on the left side. Normally a pair of these vessels joins the dorsal

vessel in the somite next posterior to the one in which the most posterior

"hearts" are found.

specimen's of helodrilus caliginosus 125

Helodrilus roseus (Savigny)

The following description is based on transverse sections of the first

twenty-three and twenty-four somites and on frontal sections of the

ventral half of somites 24 and 25 to 35 and 37. The fact that the sperma-

thecal pores of this worm are located near the mid-dorsal line eliminates

from consideration all species of this part of the country except H. foetidus

and H. roseus. Since this worm does not have the bands of color which

are characteristic of H. foetidus, it is assumed to be an abnormal specimen

of H. roseus.

External Characters. — On account of the doubling of certain somites

on one side and not on the other the somites have been numbered on both

sides independently. Posterior to 12 the left half of each somite bears a

different number from that of the right half. The relative positions of

the various organs are shown in the accompanying diagram. The location

of the diflFerent organs indicates a probable doubling on both sides of most

of the somites in the anterior part of the worm.

Four somites, 13, 34, 39, and 66, are double on the left side; and 100

and 111 are double on the right side. The total number is 162 on the

right side and 164 on the left. The maximum number recorded for a

normal worm of this species is 150. The setae bear the usual relations

to each other. Setae c and d on 16, 18, 21, 22, and 23 on the left side, on

20 and 21 on the right side, and setae a and b on 16 on the right side are

about twice as long as the ordinary setae. Glandular swellings surround

these setae. In normal worms setae of one or more bundles of 9, 10, 12, or

13 may be similarly modified. The clitellum, which is not very pro-

nounced, is saddle-shaped and does not include the ventral setae. It is

developed on fifteen somites, while in normal specimens it usually includes

but eight. These somites are identical for both sides, although the somite

numbers differ on the right (45-59) and left (48-62) sides. Each of the

tubercula pubertatis is divided into two distinct areas. The one on the left

side includes a smaller part which extends over the posterior two thirds

of 54 and the anterior one half of 55, and a larger part which extends from

the posterior two thirds of 56 to about the middle of 59. Of the one on

the right side, the larger part is more anterior reaching from the posterior

two thirds of 52 to the middle of 55; and the smaller part extends from

the anterior margin of 57 to the middle of 58. In the normal worm the

ridge of each side is continuous and extends over but three somites, 29-31.

Glandular swellings are present surrounding setae ab on somites 27

and 28 of both right and left sides of the worm. On external examination

there appeared to be a pore in the middle of each swelling, but a study of

sections of this region showed but a single pore on each side situated on

the anterior of the two glandular swellings. These are the spermiducal

pores, present on 27 of each side. In a study of the sections two pairs

126 BESS R, GREEN

of oviducal pores were found on 24 and 25 of both right and left sides.

These pores are close to setae ab and are very inconspicuous. The sper-

mathecal pores of the right side are present on 16/17-19/20, and those

of the left side are on 15/16 and 17/18-19/20.

Internal Characters.— Evidence of the doubling of somites is shown

by the presence of twice the normal number of spermaries, spermathecae,

and ovaries, and also by the large number of "hearts" that are found in

this specimen (Figs. 1 and 2). All four spermathecae of each side are

normal in form, and their pores are situated in the usual position near the

mid-dorsal line. Spermaries and spermathecae of the same side are

present in the same somites, 17-20 on the right side and 16, 18, 19, and 20

on the left. This is the usual relation of these organs. This relation is

shown very clearly on the left side; where the first spermary is separated

by one somite from the second, and the spermathecae of that side are

likewise separated. Another striking relation is shown in the development

of lateral pouches from the calciferous gland. Normally a pouch develops

on either side of the oesophagus in the somite in which the most anterior

pair of spermaries develops. In this specimen the first spermary on the

left side is in 16, and a calciferous gland pouch is also present in that

somite. The second spermary, the first of a series of three, is in somite

18, and a calciferous gland pouch is also present in that somite. On the

right side, the spermaries are present in somites 17-20, and a pouch is

found in the somite with the most anterior spermary. A third relation is

shown by the relative positions of the sperm sacs and the spermaries.

It is usual in this species for two sperm sacs to develop from the septum

which bears the second spermary, one from the anterior face and one

from the posterior face. This relation exists in this specimen in connection

with the' septum numbered 18/19 on the left and 17/18 on the right.

This septum bears the second spermary on the right side and the second

of the series of three on the left side. The spermiducal funnels show

normal relations to the spermaries and sperm ducts. Two well developed

ovaries are present on both sides, each having the usual relations to an

oviducal funnel, ovisac, oviduct, and oviducal pore. The ovaries on each

side are situated in the third and fourth somites posteriad of the somite

which contains the most posterior spermary of that side.

Eleven "hearts" are present on each side of the alimentary canal in

somites 10 to 12 inclusive. The "heart" in somite 12 on the left side is

very small. The most posterior "heart" on each side is in the same somite

with the most posterior spermary of that side. This is the usual relation

of these organs. A lateral longitudinal vessel unites with the dorsal

vessel on either side in the usual position, which is in the somite ne.xt

posteriad of the one in which the most posterior "heart" is found.

SPECIMENS OF HELODRILUS CALIGINOSUS

127

w

Iv

10

s

15

If

JL

SI

^

v:

IS

Cd 5^

-ihC-Sl.

1

fc^-

75^

■pg

St

■ -- ss

-- sd

-- OS

- - op

- -sp

10

20

II

I!

II

--P9

Fig. 1. Hclodrihts roseiis, abnormal specimen: dv, dorsal blood vessel; H, "heart";

Iv, lateral longitudinal vessel; o, ovary; os, ovisac; pg, pouch of calciferous gland; s, spermary;

sd, sperm duct; sp, spermiducal pore; ss, sperm sac; st, spermatheca.

Fig. 2. Hclodrihts roseus, normal specimen: op, oviducal pore. Other letters as for

figure 1. This diagram also represents the arrangement of the organs in a normal HelodrUus

caliginosus Irapezoides, with one exception. In the latter species the spermathecae are in-

cluded within the septa and do not extend into the cavities of the somites.

Fig. 3. HelodrUus caliginosus irapezoides, abnormal specimen: op, oviducal pore Other

letters as for figure 1.

128 BESS R. GREEN

A comparison of this specimen with a normal one naturally leads to

the assumption that for some reason, not yet obvious, there has been

some disturbance in developmental processes which has led to the develop-

ment of two somites with contained organs from each of most of the

units of developing tissue which would normally give rise to a single somite.

DEPARTMENT OF METHODS, REVIEWS, ABSTRACTS,

AND BRIEFER ARTICLES

A STUDY OF THE STABILITY OF STAINING SOLUTIONS

By

F. L. Pickett

State College of Washington

Very early in the writer's work in plant histology, he was confronted

with the statement that most staining solutions would change their

characteristics with age and that many would become entirely useless

within a period of a few months. With such conditions true, the duplication

of specific staining results would be impossible except through the use

of freshly prepared solutions. Even the use of some old and reliable

solutions would be out of the question. In many cases the careful com-

parison of methods of staining is of importance. It is often desirable to

exactly duplicate a process of staining. Individual workers find it burden-

some or even impossible to prepare stains anew for each new piece of work,

or even to prepare new solutions two or more times within a year. A

study of the more common stains and their solutions suggested no reason

for their breaking down, so a program of experimental work was planned

to determine whether or not such a breaking down was necessary, and if

so what was its cause. The results of the work have been so gratifying

that it seems wise to pass them on to other workers.

Most of the solutions were prepared in 1912. Extreme care was

used in cleaning all glass ware and utensils used in the work. Gruebler's

dyes were used, only chemicals of a reagent grade, chiefly Merck's Blue

Label, were used, and only carefully distilled water was used. The solu-

tions were stored in glass-stoppered bottles. No attempt was made at

any time to produce other than normal storage conditions. The bottles

were kept in an ordinary cupboard in a laboratory room where a wide

temperature range (5°-40°C) prevailed. From time to time the bottles

were opened for withdrawal of solutions for individual and class use. No

special seal, other than well ground stoppers, was used.

The dyes and chemicals used in the early preparation of solutions

were those considered as of a high grade at that time. There has not yet

elapsed a sufficient period for a positive statement as to the action of

dyes produced by American manufacturers since the war period, but the

writer's experience within the available time has given promise of results

129

130 F. L. PICKETT

fully as satisfactory or even better than the results obtained with Gruebler's

products. Many brands of tested reagent chemicals have also proven

entirely satisfactory.

It is the writer's opinion that most, if not all, of the trouble in the

keeping of common staining solutions is due to carelessness in preparing

the solutions, to the use of poor dyes and chemicals, or to the introduction

of foreign matter in water or from poorly cleaned utensils and containers.

The following annotated list gives the results obtained by the writer

from the storage of various staining solutions through a considerable

period. Where no date appears, the solution was prepared in 1912.

Brazilin, (Saturated solution of crystals in absolute alcohol.) — Grown quite dark brown, but

far more vigorous than when fresh as used in various formulae as for Haematoxvlin.

Carbol-Fuchsin, Ziehl-Xielson. — Filtered repeatedly to remove slight precipitate, but clear

and vigorous as ever. (1904.)

Borax-carmine, Grenacher. — As good as new in every way, and more quickly staining

Borax-picro-carmine, Baumgarten. — Just as new.

Ammonia-carmine, Beal. (Clark, Practical Microscopy, 230.) — Just a? new.

Ammonia-carmine, Gerlach. (Clark, supra, 230.) — Just as new.

Chloral-carmine, A. Meyer. — Just as new.

Litho-carmine, Orth. — Just as new.

Para-carmine, P. Majer. — .\lways a vigorous and beautiful stain. Increases vigor and in-

tensitv with age.

Carmalum, P. !Ma,\er. — Has had to be filtered several times, and is noticeably weakened.

(1904.)

.\mmonia-cochineal. Pickett. (Science, Oct. 11. 191 2 1 — Considerable precipitate on sides of

bottle, but solution clear and vigorous.

STABILITY OF STAINING SOLUTIONS 131

Congo Red. (Equal parts of alcohol and saturated solution of Congo Red in aniline water.)

— Just as new.

Cyanin. (1 gm. Cyanin dissolved in 100 cc. alcohol to which 100 cc. aniline water is then

added.) — Became almost inactive in 1921.

Fuchsin. (Equal parts of alcohol and saturated solution of Fuchsin in aniline water.) — Just

as new. (1913.)

Gentian Violet (Saturated solution in clove oil.) — !More vigorous than when new.

Haematoxylin (Saturated solution in absolute alcohol.) — Has become very dark brown in

bottle about half full. Used in making up Delalield, Erlich Acid, and Kleinenberg

solutions it hastens "ripening" greatly, so such solutions may be used within a few days.

Distinctly more valuable than fresh material.

Haematoxylin, Erlich Acid. — Has become a very dark crimson color. Heavy precipitate on

walls of bottle, but solution far more vigorous than when new. Tends to precipitate in

alcohol more than a fresh solution. (1909.)

Haematoxylin, Grenacher. — Filtered several times and considerably weakened. (1904.)

Haematoxylin, Delafield. — Precipitate on bottle sides. Weakened somewhat since 1920.

Haematoxj'lin, Kleinenberg. — As new, with slight precipitate on sides of bottle.

Magdala Red, (1% of dye in alcohol.) — Just as new.

Methyl Blue, (Equal parts of alcohol and filtered solution of 2 gm. Methyl Blue in aniline

water.) — Slight precipitate. Otherwise as new.

Orange G., (Saturated solution in clove oil.) — Just as new.

Saffranine I., (Equal parts of saturated solution of alcohol-soluble Saffranine in alcohol, and

aniline water.)

Saffranine IT., (Equal parts of saturated solution of alcohol-soluble Saffranine in alchol, and

of water-soluble Saffranine in aniline water.)

132 F. L. PICKETT

Saffranine III., (Same as 11. but with distilled water instead of aniline water.)

Safifranine IV., (Equal parts of alcohol and saturated solution of water-soluble Saffranine in

aniline water.)

All Saffranine solutions are clear and vigorous as when new.

MODERN MICROSCOPY, A HANDBOOK FOR BEGINNERS AND

STUDENTS. By M. I. Cross and Martin J. Cole. Fifth Edition.

Revised and rearranged by Herbert F. Angus. With Chapters on

special Subjects by various writers. Chicago. Chicago Medical

Book Co., 1922. X +315 pp.

This book is written for the beginner in microscopy in England or,

judging from certain portions of it, one might perhaps say London. In

looking through its pages one might well infer that the manufacturing

opticians of London were the only ones in the world. Nowhere in its

pages is there a reference to the products of the great optical works of

Zeiss, Leitz, Reichert and others in Europe or of Bausch and Lomb or

Spencer Lens Co. in America. As a further indication of the public for

which the book was written Appendix III, Microscopical Societies and

Clubs might be cited. Here four societies and clubs are discussed, of which

three are located in London and the fourth in Manchester. The societies

in the provinces are dismissed in four lines while the societies in the re-

mainder of the world get no attention whatsoever. If the user of the

book wall, however, overlook these deficiencies and will provide himself

with the catalogues of the dealers and manufacturers of this country

he cannot help finding in it much useful information.

The book is divided into three parts which will be reviewed in order.

Part I includes eleven short chapters and three appendices. It deals with

the microscope and accessories, their construction, and use. The descrip-

tions are characterized by their brevity and clarity of expression, and are

well adapted to the needs of the beginner who might be overwhelmed

by technicalities if the subject was fully presented. Many more ex-

perienced workers could read portions of this part with profit.

Part II — The Microscope and the Scientist — comprises seven chapters,

of which three deal with the microscope in medicine and one each with

the microscope in histology, geology, engineering, and agriculture. The

first four chapters of this part give some directions as to methods but the

remaining three are chiefly descriptive. All of these chapters are of

value to the beginner in that they show many of the applications of the

microscope to certain important fields of science. The treatment of each

field is necessarily incomplete. Nevertheless, it is sufficient to give a

viewpoint to the user of the book and to demonstrate that there are many

unsolved problems requiring the use of the microscope in their solution.

Part III — The Microscope and the Naturalist — with its introduction

and six chapters is very useful to the beginner who is largely self-trained,

133

134 GEORGE R. LA RUE

or to the worker who is trained in a very narrow held. The introduction

by Wilfred Mark Webb strives to create in the amateur a desire to investi-

gate the things about him and it hints at the wealth of objects for study

in any environment. The chapter on Pond Life by the late C. F. Rousselet

has admirable notes on methods of collecting, preliminary examination

and keeping, apparatus for microscopical examination, preserving and

mounting. There is also a collector's calendar which doubtless is of

considerable value in England and is suggestive of what might be done

for various regions of America. In subsequent chapters, each by a special-

ist, a somewhat similar treatment is accorded to the fresh-water mites,

the foraminifera, mosses and liverworts, and mycetozoa. There is a

final brief chapter by M. J. Cole entitled 'Mounting Common Objects.'

This, together with the chapter on 'The Microscope in Histology' and the

three chapters on 'The Microscope in Medicine,' gives the beginner,

especially if he be inclined toward biology, considerable practical informa-

tion.

The book is well indexed. Typographical errors are not abundant

and are not serious. One notes particularly, however, the word 'Ashe'

on page 63 misprinted for 'Abbe.'

The reviewer believes that the authors and publishers are to be com-

mended for having made available to amateur and student microscopists

the information contained in this book. He feels that, like the amateur

naturalists, the amateur microscopists have been altogether too much

neglected by those who have had special training and wide experience in

the various fields of research involving the use of the microscope. At the

same time he feels that this book might have been made very much more

useful for its American public had portions of it been rewritten with this

public in mind.

George R. La Rue

TRANSACTIONS ' '^

OF THE

American

Microscopical Society

Organized 1878 Incorporated 1891

PUBLISHED QUARTERLY

BY THE SOCIETY

EDITED BY THE SECRETARY

PAUL S. WELCH

ANN ARBOR, MICHIGAN

VOLUME XLII

Number Three

Entered as Second-class Matter August 13, 1918, at the Post-office at Menasha,

Wisconsin, under Act of March 3, 1879. Acceptance for mailing at the

special rate of postage provided for in Section 1103, of the

Act of October 3, 1917, authorized Oct. 21, 1918

BIIjE OloUigiaie ^mbb

GEORGE BANTA PUBLISHING COMPANY

MENASHA, WISCONSIN

1923

TABLE OF CONTENTS

For Volume XLII, Number 3, July, 1923

A Study of the Movements of Entoproctan Bryozoans, with two plates, by W. A. Hilton 135

Observations on the Life Cjxle of Davainea proglottina in the United States, with two

figures, by A. C. Chandler 144

The Egg Laying Habits of Haminea virescens (Sby), with three figures, by A. Richards . . 148

Department of Methods, Reviews, Abstracts, and Briefer Articles

Note on the Occurrence of the Marine Cladoceran Evadne tergestina in Southern

California, by Helen E. Murphy 155

Methods of Culturing Tubificidae in the Laboratory, by E. F. Powell 155

A New Method for Whole Mounts, by W. K. Bowen 156

TRANSACTIONS

OF

American Microscopical Society

(Published in Quarterly Instalments)

Vol. XLII JULY, 1923 - No. 3

A STUDY OF THE MOVEMENTS OF ENTOPROCTAN

BRYOZOANS

By

William A. Hilton

Department of Zoology, Pomona College, Claremonl, California

When the bits of rock or shell covered with living Barentsia or Myosoma

are lifted from the water, a very marked movement of all or nearly all

the zooids is evident. In the case of the first of these, the characteristic

incomplete or partial rotations of each individual are seen to be caused

by contractions of the swollen muscular base of each stem. With Myosoma

the movements differ slightly because the whole stalk is muscular and

flexible. The observations in this paper are chiefly upon Barentsia gracilis

Hincks which is common at Laguna Beach. The species determination

was made by Dr. Robertson, whose extensive papers on Pacific Coast

Bryozoans are the chief source of information for the identification and

distribution of western forms.

The questions which suggested themselves at once when these forms

were examined were; what are their movements, how are they stimulated

to move, and what relations do the nervous system and sense organs bear

to these activities.

The most evident movements of Barentsia are those which carry the

body with its tentacles through a wide arc by the contractions of the

muscular base of the long stalk. These movements tend to describe a

complete circle but if obstructions are met with, the rotation ceases and

may not be continued unless the zooid is further stimulated. Movements

of the sort just mentioned tend to persist in one direction when once

started, although at other times the contractions may cause a rotation in

the other direction. The next most marked change is seen in the contrac-

tion of the tentacles. When a mass of the colony is lifted from the water

or otherwise very strongly stimulated, the tentacles contract and remain

in this condition for some time even though the other activities are as

marked as ever. The body of each zooid is attached to the stalk by a

small area of flexible tissue and under certain conditions this permits of a

slight rotation of the body upon the stalk.

135

136 WILLIAM A. HILTON

The work of the past upon this group of Bryozoa gives little hint of

their behavior or the nature of their activities but many writers have

some suggestion of the structure and arrangement of the nervous system,

and for that reason a brief review of the literature follows.

Van Beneden, 1845, considers PediceUina but gives Httle indication

of the nervous system. Kowalevsky, 1867, considers the development.

Uljanin, 1869, gives the position of the central ganglion in th's genus.

Nitsche, 1875, shows the general position and chief branches of the central

nervous system in PediceUina. Salensky, 1877, indicates the general

location of the ganglion in Loxosoma, and Harmer, 1885, gives the most

complete early account of the nervous system. In Loxosoma he describes

a dumb-bell shaped ganghon with nerve cells on the outer surface of the

central fibrous part. There are many bipolar sense cells, especially in the

tentacles which send their fibers into the brain. Foettinger, 1887, repre-

sents the brain of PediceUina by two more or less separate lobes. Several

pairs of nerves pass from the ganghon. Seeliger, 1890, gives the develop-

ment and position of the nervous system. Davenport, 1893, shows the

position of the ganglion in UnaleUa. Nickerson, 1901, in L. davenporti,

describes the brain as just in front of the intestine and above the stomach.

It is elongated transversely. From each end two bundles are given oflF,

one on each side to pass to the lophophore. Sensory bristles were seen

on the tentacles. Stiasny, 1905, shows the ganglion of PediceUina but

with no detail. Retzius, 1905, shows sensory nerves on the surface of

PediceUina. These sensory cells bear bristles and are connected with

nerve strands which form a wide network of fibers. Sensory cells were

marked in the tentacles. Assheton, 1912, found the nervous system in

two species of Loxosoma. The brain and chief branches are figured, and

sense cells are mentioned on the hypostome, lophophore and tentacles.

In Barentsia and Myosoma the central ganglion is small. Its connec-

tion with other parts was not traced, but after trials during several summers

sense cells were demonstrated in the tentacles, and especially at the tip of

the stem where it joins the body of the zooid. Sense cells and a plexus

of nerve fibers or a nerve net were demonstrated, especially forming a

network in the unswollen part of the stem. Bipolar nerve cells of the stem

were demonstrated sending their fibers into the sense pores or the little

pits in the cuticle of the stem. These pits occurred only on the unswollen

parts of the stem. Methylene blue was used to show the sense cells and

the nerve net.

In Myosoma, instead of sense pits the stems were covered with hollow

sensory hairs much like those so common in arthropods, but in this form

no demonstration of the sensory cells was made. Although there are no

sense cells in the swollen base there is a very marked nerve net under the

cuticle in Barentsia.

THE MOVEMENTS OF ENTOPROCTAK BRYOZOANS 137

So far as I have been able to determine, there is no account of the

activities of Barentsia and no experiments to determine their control.

In Loxosoma the movements are quite different, partly because of their

not being colonial forms and partly because of their different form and

structure, so no time will be taken to discuss them further. Barentsia has

a creeping stolon which is often hidden by other growths, but sometimes

it may readily be seen upon the surface of a rock or a bit of sponge and it

is evident that there is great variation in the way in which the individuals

of the loosely formed colony are linked together. Each individual of a

chain or network has its own part of the stolon cut off from the rest by

well-marked partitions across the stem.

In summarizing the general effects of various stimuli upon the move-

ments of Barentsia, the two minor movements may be dismissed with a

brief statement. The tentacles do not withdraw themselves unless the

stimulus is quite severe or sudden. They may be withdrawn when the

animals are removed from the water, they contract when touched rather

directly and they do not change much in relation to light or chemicals

either when expanded or contracted.

The rotations of the body on the stem at the point where the stem

joins the body are not very marked at any time, but they seem to be more

evident with freshly obtained individuals that are strongly stimulated

especially by touch or jar, although they were occasionally seen to move

in this way when no exciting cause was evident.

The response which is most marked and which is chiefly considered in

the following account is the rotation due to contractions of the swollen

base of the stem or stalk. The maximum reaction is a movement of 360°.

Freshly obtained specimens tend to perform this greatest movement and

alth(^gh after a time there may not be quite so great a swing, yet the

members of the colony remain active for a long time. This activity is not

primarily due to stimuli coming to the body as is shown by entirely re-

moving it from the stem, for then the rotations are as marked as before.

Tactile stimuli were used in large part because these could be more de-

finitely localized than any others. It was found that the most sensitive

areas were first the tip of the stem where it joins the body, and next the

stem or stalk. Even the swollen base of the stem is more sensitive than

the body even though no sensory endings were demonstrated in it. As a

jar is one of the most powerful of the stimuli, it may be that this explains

the reason why the fleshy base may be stimulated to cause the movement

and why the body because of its flexibility does not transmit the jar so

well. Besides tactile stimuli which have always an element of jar, currents

in the liquid are also effective. There is but slight response to light either

continuous or intermittent. They seem to move a little more in the

brighter light. There is also but slight response to heat and if the tempera-

138 WILLIAM A. HILTON

ture is gradually increased they may die without movement. Rather

weak acids cause a sharp response but stronger ones or strong poisons

may kill them quickly with little evident reaction.

It would seem probable that under natural conditions where the colonies

are hidden in dark crannies of rock surfaces that stimuli in the nature of

contact, pressure, or currents would be the ones to bring about the marked

movements which may be something in the nature of avoiding or freeing

reactions. The violent movements induced by removing the zooids from

the water may be due to the unwonted weight brought upon the stem by

the rather heavy bodies.

As a jar seems to be the most effective in bringing about a response

when the zooids are touched, it may be that the tactile stimuli employed

have an element of jar and either the jar or a combination of effects from

jar and touch both bring about the responses. The creeping stolon is very

sensitive to the least touch as is shown by the activity of one or several

stems, but here even more than on the stems or their swollen bases is it

difficult to discriminate between jar and touch. However, living stolons

seem to carry the responses better than dead portions. A nerve net was

demonstrated in the stolons but it was impossible to be sure that the nerve

actually crossed from one member of the colony to another across the

partition in the creeping stem. If the stolon is stimulated near where

several parts join there may be a very active response in the way of rota-

tion in a number of nearby zooids. Once when a stem was cut through

there seemed to be a shock to a number of zooids for they did not respond

so well for a time when stimulated through this cut portion.

In most cases tactile stimuli were tried by means of a needle point.

During the observations zooids were kept in fresh sea water under as

normal conditions as possible. In most cases the stems or muscular bases

of the stems were touched, but in some expanded individuals the tentacles

were touched. The usual response was a shght contraction of the tentacles

stimulated near or on that side and a slight movement of the body on

the stem. Repeated stimuli cause the tentacles to draw in more. If the

center of the tentacles is touched there are more violent movements of

the tentacles and of the body but the stem may not rotate unless the touch

is quite heavy. Small regenerating bodies are more sensitive and the stems

may be induced to rotate when these are stimulated. When the body is

removed, the cut end of the stem is especially sensitive and when touched

the stem rotates violently.

When a zooid is stimulated to begin a movement further stimuli even

from several directions do not as a rule change its direction or movement

for some little time.

When an individual is touched lightly it may not move; if light stimuli

are continued it may begin to move after three or more light stimuli.

THE MOVEMENTS OF ENTOPROCTAN BRYOZOANS 139

If light stimuli are continued it may move away rather more violently.

When a specimen has been caused to move a number of times it may

become fatigued and not move again for a period which differs in different

individuals. Small specimens often respond more quickly than large ones.

When a zooid is strongly stimulated the effects may be carried to

more distant zooids, and a whole chain or a large group may be caused

to move by the movements of one. A stimulus to one zooid may not pro-

duce movement in it but others more distant may be affected. In some

cases there seems to be a blocking of an impulse when intermediate zooids

do not respond as for instance when the intermediate one is dead or

fatigued. A stimulus to a moving zooid apparently may not increase its

activities but more distant individuals may start to move although they

have just ceased moving. In this apparent blocking of impulses there is

quite a little chance for error in interpretation because a slight jar or lack

of it may be hard to determine in individual cases. In respect to fatigue

there seems to be little question that the results are as indicated. In regard

to the apparent summation of stimuli before a response, it might be urged

that the stimuli were not all even and the last one given was heavier than

the rest but careful checking of this seems to justify the result obtained.

Various anaesthetics were used on the living specimens. Chloral hy-

drate or Chlorotone were used until there was no longer any response.

The body and tentacles were first to lose their sensitiveness and the last

to regain it.

General Conclusions

1. Barentsia has four chief motor activities:

(a) Movement of the zooid as a whole through rotation of the stem

caused by contractions of its fleshy base.

(b) Movement of the body on the stem.

(c) Movement of the tentacles, contraction and expansion.

(d) Movement of external and internal cilia.

2. As suggested by both experimental and anatomical methods there

are three chief centers of motor control: (a) The ganglion of the body for

the control of the tentacles, (b) The plexus or nerve-net where the body

joins the stem. This is for control of the movements of the body on the

stem. Apparently also, here is a sensory area for receiving and carrying

down the stem sensations or movement from the body. Possibly this

is the chief connection the body has with the stem, this rather indirect

one. In any case the connection is not marked judging from the experi-

ments in stimulating the body, (c) The sensory pits, bipolar cells con-

nected with them and the nerve-net of the stem. Sensations are received

here and conducted down the stem. There is little or no evidence of their

going in the other direction or affecting the body or tentacles in any way.

140 WILLIAM A. HILTON

(d) The very marked nerve plexus in the swollen base of the stem which

receives fibers from the stem above or from the plexus of the creeping stolon

and responds to impulses coming over these fibers. The swollen base may

receive impulses in the way of jar or pressure directly, but because of the

thickness of the cuticle and the lack of special tactile organs it seems not

to be so sensitive to light touch as the stem or stolon. It seems to be the

center for the control of the larger movements of the stem, (e) The creep-

ing stolon marked off by partitions between zooids. This has sense pits

and a nerve plexus similar to the stem above the swollen base.

3. There are then three centers of control: (a) The ganglion in the

body for the tentacles, (b) The first part of the stem especially for move-

ments of the body on the stem, (c) The swollen base which is not only a

contractile organ but also contains the nerve plexus which controls the

larger movements of the stem.

4. It seems possible to fatigue the zooids by continuous movements.

5. Fatigued zooids may block the pathway of a stimulus.

6. Stimuli pass from zooid to zooid along the stolon.

7. Stimuli to the tentacles may cause them to contract without affect-

ing other parts. Rotations of the zooid may not affect the condition of the

tentacles if the body does not touch anything.

8. Weak stimuli which do not individually produce a response may be

summed and cause a reaction.

Bibliography

ASSHETON, R.

1912. Loxosoma loxalina and L. sultans, two new species. Q. Jour. Mic. Sc, vol. 58

N. S., pp. 117-143, pi. 6-7, 4 text f.

Davenport, C. B.

1893. On Unatella gracilis. Bull. Mus. Comp. Zool., vol. 24, pp. 1-44, 6 pi.

FOETTINGER, A.

1887. Sur I'Anatomie des Pedicellines de la cote d'Ostende. .\rch. dc Biol., vol. 7, pp.

299-329, pi. 10.

Har.mer, S. F.

1885. On the Structure and Development of Loxosoma. Quart. Jour. Mic. Sc, vol. 25,

pp. 261-337, pi. 1-3.

1886. On the Life-history of Pcdicellina. Q. Jour. .l\[ic. Sc, vol. 27, pp. 239-263, pi.

21-22.

Hatschek, B.

1877. Embryonalentwicklung und Knospung der Pcdicellina echinata. Zcit. f. wiss.

Zool., Bd. 29, pp. 502-562, pi. 27-30, 4 wood cuts.

NiTSCHE, H.

1869. Beitrage zur Kenntniss der Bryozoen. II Ueber anatoniie von IVdicellina echin-

ata Sars. Zeit. f. wiss. Zool., Bd. 20, pp. 13-36, pi. 2-3.

NiCKERSON, W. I).

1901. On Lo.xosoma davcnporti sp. nov. Jour. Morph., vol. 17, pp. 351-380, pi. 22-23.

THE MOVEMENTS OF ENTOPROCTAN BRYOZOANS 141

Prouho, H.

1891. Contributions a Fhistiore dcs Loxosoines. Etude sur le Loxosoma annelidicola.

Arch. Zool. exper., 2 e ser. t. 9, pp. 91-116, pi. 5.

Retzius, G.

1905. Das sensible nervensystem dcr Bryozocn. Biol. Unters. N. F., Bd. 12, pp. 49-54,

pi. 5.

Salensky, M.

1887. Etudes sur le Bryozoaires endoproctes. Ann. des. sc. Nat. 6 ser. Zool.,Bd.5, pp.

1-60, pi. 12-15.

Seeliger, O.

IStO. Bemerkungcn 2ur Knospen-Entwickelung der Bryo^ioen. Zeit. f. wiss. Zool.,

Bd. 49, pp. 169-208, pi. 9-10, 6 figs.

V'erworn, M.

1887. Beitrage zur Kenntniss der Susswasserbryozoen. Zeit. f. wiss. Zool., Bd. 46, pp.

99-1.^0, pi. 11-12.

142

WILLIAM A. HILTON

S

Plate VI

Fig. 1. Sketch of a single preserved specimen of Harentsia. xl7.

Fig. 2. Sketch of a single preserved specimen of JM\osoma. xl7.

Fig. 3. Zooid end of Barentsia. Nerve ganglion in dark. x35.

Fig. 4. Zooid end of Myosoma. (langlion in dark. x35.

Fig. 5. Section of the central ganglion of Barentsia. xl50.

THE MOVEMENTS OF ENTOPROCTAN BRYOZOANS

143

Plate VII

Fig. 6. Section of the stem and sense hairs of Myosoma. xl50.

Fig. 7. Section of the stem of Barentsia. xl50.

Fig. 8. Optical section of base of zooid and stem of Barentsia, sliowing sense cells stained

with methylene blue. x35.

Fig. 9. Base of stem of Barentsia stained with methylene blue to show sense pits and

nerve network. x30.

OBSERVATIONS ON THE LIFE CYCLE OF DAVAINEA

PROGLOTTINA IN THE UNITED STATES

By

Asa C. Chandler,

Biological Laboratory, Rice Institute, Houston, Tex.

In the course of parasitological examinations of slugs, Agriolimax

agrestis, made on January 23rd, one specimen out of about 25 collected

from a chicken yard contained very large numbers, 150 or more, of tailless

cysticercoids which morphologically resembled the scoleces of Davainea

proglottina. The heart-shaped cysticercoids measure from 2.30 x 160/i to

160 X 140 ju and possess a gelatinous wrinkled outer coat, varying in average

thickness from about 14 to 30/x, but always thicker at the posterior end.

The variation in size of the cysticercoids is due principally to the thickness

of this outer coat. The wrinkled or corrugated surface reminds one some-

what of the gelatinous coat of an Ascaris egg. A rather heavy fibrous wall

delimits the heart-shaped inner body of the cysticercoid which contains

large numbers of very refractile calcareous corpuscles averaging about lO/x

in diameter. The withdrawn scolex measures about SO^u in diameter, but as a

rule its outline is very indistinct, being obscured by the calcareous granules.

The single rostellar circlet of about 75 to 90 booklets is very distinct; the

booklets measure about 8^i in length. The suckers themselves are readily

distinguishable by the single circlet of hammer-shaped booklets.

Some of the cysticercoids were found free in the body cavity of the

slug, but many of them were imbedded in the tissues, particularly in the

walls of the alimentary canal.

Portions of the slug containing numerous cysticercoids were fed to two

hens, each about two and a half years old, which by fecal examination were

found to be free from any species of Davainea, although the eggs of another

cestode, which was subsequently found to be Amoebotaenia sphenoides,

were found. These birds came from a flock of six hens which had been

kept away from other poultry for over a year, and two other members of

the flock which were examined at other times for parasites were also found

to be free from any species of Davainea.

On February 12th, twenty days later, the droppings of the hens were

examined and found to contain active proglottids of Davainea proglottina

in considerable numbers. It is probable that the worms had matured

some time before, inasmuch as Grassi and Rovclli (1889, 1892) found that

the adult stage was reached in chickens eight days after ingestion of in-

fected slugs. Grassi and Rovelli's work was not at hand at the time, and

144

THE LIFE CYCLE OF DAVAINEA PROGLCTTINA

145

it was not realized that development would take place so rapidly. One of

the hens was killed and found to contain about 50 specimens of Davainea

proglottina and numerous individual proglottids. The droppings of the

other hen were examined at intervals of three or four days to determine

how long the active proglottids would continue to be voided. On Feb. 21.

only a single proglottid could be found in a night's droppings, two were

found on Feb. 24, and none at any time after that date, which would indi-

cate that in birds of the age of these hens the infection is not of long

duration.

O.l mm.

Fig. 1. Cysticercoid of Davainea proglottina from naturally-infected Agriolimax

agrcstis

A number of living proglottids were fed, on lettuce leaves, to two

laboratory bred slugs, Agriolimax agrestis, on Feb. 12, and a number of

others were fed to four other laboratory bred slugs on Feb. 13. The latter

slugs were later found to be of another species, Limax flavus, identified by

Dr. H. A. Pilsbry, and said by him to be the first record of its occurrence in

the southwest.

One specimen of Agriolimax agrestis was examined on Feb. 22, ten

days after infection, and found to contain about twenty young larvae, in

a very early stage of development. These larvae measured from 127 x 98^

146

ASA C. CHANDLER

to 145 X 118^1 but were practically undifferentiated, showing no signs of

invagination. The embryonic hooks, about 16/x in length, were still

present, situated as shown in Fig. 2. No indication was found of a vestigial

tail, such as is described by Grassi and Rovelli. The other specimen of

Agriol'imax agrestis was examined on March 6, 22 days after infection,

and found to contain cysticercoids similar to those found in the naturally

infected slug.

Two of the four specimens of Limax flavus which were fed proglottids of

Davainea proglottina on Feb. 13, were examined on Feb. 27, 14 days after

infection. One specimen was entirely negative, while the other contained a

single cysticercoid measuring 213 x 213/x. This specimen is heart-shaped,

has invaginated, and shows a retracted scolex, but there is no indication of

rostellar hooks, suckers, or acetabular hooks, nor of a vestigial tail. The

Fig. 2. Young larva of Davainea proglottina from Agriolimax agrestis, 10 daj's after

laboratory infection

calcareous granules are much smaller than in the fully developed cysticer-

coid, and the entire specimen has a coarsely granular appearance. A

third specimen examined on Mar. 3, 18 days after infection, was also

found to be entirely negative. The fourth specimen escaped.

It would appear from the above experiments that Agriolimax agrestis,

which is one of the species which Grassi and Rovelli were successful in

infecting, serves as an intermediate host in the United States as well as in

Europe. Limax flavus would appear to be highly resistant to infection,

since only a single cysticercoid developed out of hundreds of eggs fed to

three individuals, and it is not probable that this species would serve as an

intermediate host in nature.

THE LIFE CYCLE OF DAVAINEA PROGLOTTINA 147

The occurrence of Davainea proglotlina in the United States has

previously been recorded only by Ransom (1909) from Pennsylvania and

Maryland. Guberlet (1916) states that this species has not been recorded

in the United States, evidently having overlooked Ransom's records, which

are cited only in a footnote. His erroneous statement has been copied in

text books and has led to the general assumption that this species has

never been introduced from Europe. Its occurrence in such widely

separated regions as Pennsylvania, Maryland and Texas indicates a rather

wide distribution in this country.

Literature Cited

Grassi, B. B., and Rovelli, G.

1889. Embryologische Forschungen an Cestoden. Centralbl. f. Bakt. u. Parasitenk.,

5, 370-377, 401-410.

1892. Ricerche embriologische sui Cestodi. Atti Accad. Gioenia di Sci. Nat. in Cata-

nia, 4, 1-108.

Guberlet, J. E.

1916. Studies on the Transmission and Prevention of Cestode Infection in Chickens, J.

Amer. Vet. Med. Assn., 2, 218-237.

Ransom, B. H.

1909. The Taenioid Cestodes of North .\merican Birds. Bull, U. S. Nat. Mus., 69,

1-141 (p. 68).

THE EGG LAYING HABITS OF HAMINEA VIRESCENS (SBY)*

By

A. Richards

The green bubble shell, Haminea virescens Sowerby, belongs to the

opisthobranch gasteropods of the family Bullidae. The genus occurs in

suitable locations on both Atlantic and Pacific Coasts. According to

Gould it is "common in the muddy lagoons and salt ponds along the shores

of Vineyard Sound" and elsewhere in the Woods Hole vicinity. This spe-

cies may be found along the southern California coasts on tidal flats bur-

rowing into the sand at low tide.

Embryological phases of three species of this genus have been studied.

Smallwood in 1901 reported observations on the behavior of the centro-

some during maturation of the eggs and in 1904 described in detail the

maturation, fertiUzation, and early cleavage of H. solitaria. In another

paper of 1904 he discussed the natural history of the species, including

the egg lay habits. The species found in Puget Sound is H. vesicula. Its

early development was reported by Leonard in 1918, and Bovard and

Osterud include a note on the form of the egg masses in their paper of the

same year. Finally, the writer described briefly in 1922 the habits of

H. virescens obtained at La Jolla, California, and gave an account of

experiments performed on them.

The accounts of the egg masses of these three species are by no means

in agreement. For this reason it is desirable to bring together the descrip-

tions of all three, pointing out in detail the condition observed in vires-

cens.

During the summer of 1921 while working at the laboratory of the

Scripps Institution for Biological Research at La Jolla, I used the eggs of

Haminea virescens for experimental purposes. For reasons that will appear

in connection with the description of the egg masses these eggs are espe-

cially suitable for experiments in which it is desirable to maintain to the

greatest possible degree the normal environment of the egg. The animals

were brought into the laboratory and kept in a dish of running sea water

in which were stones and a quantity of sand from the tidal Hat where the

specimens were originally obtained. No difficulty was experienced in get-

ting them to produce eggs for a number of days. At length, however, they

became exhausted and a new supply had to be obtained. Whether the

eggs failed because of a natural course of events or because of the labora-

tory conditions could not be determined.

Egg laying occurs usually about the time of ihc first light in the morn-

ing. It is not improbable that the light acts as a stimulus of some nature.

* Studies from the Zooloj^ical Laboratory of the Universit)' of Oklahoma, Second Series

No. 36.

14cS

EGG LAYING HABITS OF IIAMINEA VIRESCENS 149

I was unable to observe the actual laying process, but from the known facts

for the periods elapsing during the cleavage stages and from the comparison

with similar phenomenon in other gasteropods, it is probable that egg de-

position took place in the early morning between three and four o'clock.

Light in this latitude comes during the summer months very early, for the

sun rises between 4:30 and 5 o'clock. There are of course exceptions to

the rule that egg deposition takes place this early. I have found one cell

stages at 6:20 and 6:45 A. M., and in one case when I came into the labora-

tory late in the afternoon (at 5 :30 p.m.) I found an egg mass in the four cell

stage indicating that it had been deposited perhaps four hours earlier, or

not long after mid day. Perhaps the laboratory conditions may have

been responsible for this irregularity. On the other hand, I found 12 cell

stages as early as 6:30 A.M. on several occasions and once as early as

5:40 A.M. Nevertheless, usually it appeared that the first sign of light

must have been the time of laying. Leonard finds likewise that vesicida

lays during the early morning hours. She says, "The writer has gathered

large numbers of the animals and of their characteristic egg-masses fiom

the under side of the Ulva at all times of day. Eggs in the 1 -celled stage

and apparently freshly laid were obtained most plentifully between the

hours of 5 A.M. and 9 A.M., and Haminea is frequently found in the act

of laying between these hours. However, one of the animals was found

laying as late as 11:30 A.M., and continued laying in the laboratory until

2:35 P.M."

For some animals it has been suggested that a relation of some unknown

character may exist between the times of egg deposition and the changes

of tides. My observations for Haminea are hardly in keeping with this

hypothesis. They were extended over a period of two months and showed

that the time of laying varies only as indicated above, while the tide of

course varied each month through the diurnal cycle.

The duration of the early cleavages is illustrated by one typical case

which was followed through in detail. The figures for this case are about

the average for all those studied. The observations on these eggs at 21 . 5°C

(which temperature is the upper limit of the temperature range of the sea

water in the laboratory during the summer, and the exact reading of the

thermometer on the 3rd of August when this observation was made) are

as follows:

1 cell stage first observed at 6:45 a.m.

Maturation completed at 7:45 1 i u ic •

^ ^ I 1 hr. 25 mm.

First cleavage (2 cells) completed at 9:10 J

Second cleavage constriction begun at 9:45 I n •

o , , X ) 51 mm.

Second cleavage (4 cells) completed at 10.01 J

Third cleavage constriction begun at 10:40 1 c^ •

^, . , , ° ,„ „ , r . , [ 59 mm.

Ihird cleavage (8 cells) constriction begun at 10:40 J

Fourth cleavage (12 cells) completed at 11:45 50 min.

Fifth cleavage (16 cells) completed at 12:30 40 min.

150 A. RICHARDS

On the average it appears from this and other data that the early

cleavages range in duration from eighty minutes down to forty minutes.

The longer cleavages are of course the earlier ones. The veliger stage is

reached at the end of about 48 hours. Hatching begins on the fifth day

usually and is completed in two or three days more. The percentage of

embryos that hatch is very large unless an external factor interferes.

Often I have seen 98-100% hatch, while seldom do less than 85% reach

this stage.

From Smallwood's records (1904, a) it appears that the duration of

the cleavage divisions of solitara is less than in the case of virescens. Ac-

cording to him the first polar body is given off ten to fifteen minutes after

laying and the second thirty minutes later. "The egg segments into two

cells a half hour after the second polar body has appeared" . . . "Within

thirty or forty minutes after the formation of the two cell stage, the four

cell stage is formed." . . . "After not more than thirty minutes, the

third cleavage separates the egg into two conspicuous parts, the proto-

plasmic micromeres and the deutoplasmic macromeres." . . . "The time

that intervenes between the formation of the second and third quartettes

of micromeres is the same as that for the second and third cleavage."

The structure of the egg case is the chief point in which my observations

differ from others which have come to my attention. The egg cases of the

genus Haminea are usually described as jelly like masses apparently with-

out special form. In the "Catalog of Marine Fauna (of Waters of Woods

Hole and Vicinity)," Sumner, Osburn and Cole record a note by Conklin

in reference toHaminea solitaria to the effect that the eggs are "laid in large

jelly like balls, which are fastened by stalks to the sand," and are deposited

"August 20th or earlier." Others speak of formless masses of jelly con-

taining the eggs.

Smallwood's account (1904, a) of the process in H. solitaria follows.

After describing the process of copulation which is of about fifteen minutes

duration and occurs some eight to twelve hours previous to laying, he says,

"This species lays a single gelatinous mass which is spherical, about three-

quarters of an inch in diameter. Its contents are chiefly composed of

albumen, which is secreted by the albumen gland. As soon as the albumen

comes in contact with the water it swells by the rapid absorption of water,

and thus affords a gelatinous protection for the egg. When the eggs first

leave the genital groove they are in strings; in a few hours the strings lose

their continuity and the eggs are scattered throughout the egg mass. It

would be very difficult to count the eggs in a single mass. The size of the

capsule varies considerably; as a rule those foupd on the eel-grass are about

a third less in diameter than those laid on the bottom. The egg masses

laid in the laboratory were often irregular in shape and much smaller

EGG LAYING HABITS OF HAMINEA VIRESCENS 151

than those collected from the pond. The specimens in confinement that

laid small and irregular masses, often laid a second time without a second

copulation. It takes from 40 to 50 minutes for an animal to lay a complete

normal egg mass."

The process of egg laying in H. vesicula was observed in the laboratory

by Miss Leonard, whose description is as follows:

"In the process of laying the eggs are extruded in a flat band about 1

cm. in width from beneath the right side of the mantle, and pushed back

so as to extend posteriorly. During this process Haminea is moderately

extended. At intervals definite contraction of the mantle on the right side

occurs, at which time two or more strands of ribbon are pushed out. Due

apparently to a slow spiral motion of the female while in the act of laying,

the egg-masses are spirally coiled when of sufficient length."

"Each egg is enclosed in a comparatively large capsule, containing a

transparent fluid which does not become opaque upon fixation. These

capsules are embedded in a clear, transparent, colorless or pale yellow jelly,

which is in the form of a flat band about 1 cm. in width and varying from

a fraction of a centimeter to 6 cm. or more in length. The bands are

attached by their lower border to the substratum. The eggs vary in

diameter from 50 to 90 microns and are embedded in the gelatinous band

in a continuous spiral line. The eggs when collected in their masses will

live and develop normally for several days under laboratory conditions."

"Fertilization is internal, hence the eggs when collected had already

been fertilized. None were found at a stage previous to the formation of

the first polar body, and none were dissected out of the oviduct to ascertain

whether the first polar body is formed before extrusion. No unfertilized

eggs were found, and it is a question whether any are laid. In surface

view the egg is bright yellow, with opaque yolk granules scattered thruout.

After the egg is laid the second polar body is thrown off by the formation

of a typical polar spindle."

In their list of animals yielding embryological material, Bovard and

Osterud have the following note in reference to H. vesicula. "The egg

masses are 12-38 mm. in height, and are whitish balloon-shaped bodies,

with numerous eggs enclosed in a gelatinous substance. These may be

anchored in mud or on eel grass. The masses are best collected between

five and nine o'clock in the morning to obtain the earliest cleavage stages."

I am unable to explain the discrepancy in the two accounts of the egg

mass in this species. It hardly seems however that Miss Leonard's account

can be questioned since it is based upon her own direct observations of the

animal while laying.*

* Since the above was written I have mj'self seen egg cases of H. vesicula and can substan-

tiate Miss Leonard's statements. The egg case of vesicula is in all essentials like that of

virescens.

152 A. RICHARDS

While experimenting on the rate of division in the eggs of H. virescens,

the writer kept the animals in the laboratory for several weeks and obtained

many egg cases, so that there could be no doubt as to the accuracy of the

identification of the egg masses or of their similarity to those brought from

the collecting grounds to the laboratory.

At first glance the egg cases of Haminea virescens appear like thickened

sections of ''gros grain ribbon" ranging in length from 15 to 35 mm., and

about 5 or 6 mm. broad, and of a creamy white color. Even when the cases

are taken in open water and have become covered with foreign particles

they have never given me the impression of a formless mass. Usually one

edge of the ribbon is attached to a small stone while the other floats freely

in the water. The attachment is made in such a manner that the ends of

the ribbon almost form a completed circle. In this position the eggs

develop until hatching is completed when the jelly disintegrates and the

entire structure falls to pieces.

The structure of the egg case is of a complicated pattern, as shown in

the accompanying figures. The eggs appear to be extruded in a string of

tough gelatinous material which becomes surrounded by the matrix jelly

forming the body of the ribbon. Inside of this gelatinous string the eggs

are laid alternately so that the appearance is that of a double row of eggs.

The string itself is accurately placed in the form of a flattened spiral the

loops of which are not formed of back and forth folds as they first appear,

but are so arranged that the opposing loops are compressed against each

other. This produces the effect of a cross striated thick ribbon.

In one ribbon, that may serve as a typical case, the number of loops

producing the striations was counted and found to be 242. In a single

length the number of eggs ranged from 80 to 104, averaging 90. There

were then in this egg case 21,780 eggs. Probably the number of eggs

deposited at one time under favorable conditions ranges from 15,000 to

25,000; at least the data for other normal cases indicate such numbers.

When conditions are not so favorable a much smaller number of eggs, even

as low as 5,000 is deposited at one time.

One of the most striking facts in regard to these egg cases is that the

eggs are always in the same stage of development. Uniformly I found that

the entire lot of eggs in a single egg case were in the same stage of mitotic

division. It is a remarkable condition that 20,000 eggs should be laid in

as complicated a manner as this so nearly at one time that all would divide

simultaneously. In these cases however, it was not possible to distinguish

by any difference in development which end of the mass was produced first.

In other cases where enormous numbers of eggs are shed in the water before

fertilization takes place, it is rare to find 100*;^ of the eggs developing, but

this is the rule in the cleavage stages of Haminea virescens.

Thus it appears that in this genus two entirely distinct modes of

EGG LAYING HABITS OF HAMINEA VIRESCENS

153

egg protection are to be found. In solitaria the egg case is a spherical

gelatinous mass of three-quarters of an inch in diameter. In virescens the

mass is in the form of a narrow ribbon which may be an inch and a half in

length with the eggs placed in very regular order. Vesicula agrees with

virescens in the general structure of the egg mass. It is not improbable

that the statements of some that the egg cases in this genus are irregular

formless masses of jelly are based upon incomplete data.

■■"^y. ^'-34 -'"

/'^■"''"x'

Explanation of Figures

Fig. 1. Egg masses of Haminea virescens, in natural position at about natural size.

Fig. 2. Small section of the case showing the arrangement of the spiral loops.

Fig. 3. The upper ends of two loops, showing the eggs in which the spirals are ar-

ranged in double rows. Each egg is surrounded by a gelatinous covering within the egg case.

The ribbon like structure of the egg case of H. virescens very greatly

facilitates the use of these eggs for experimental purposes. The egg case

may be cut into sections one of which will serve as a control while others

may be placed in various solutions and the results noted. The eggs them-

154 A. RICHARDS

selves are not disturbed in their gelatinous coverings and the effects of the

experiment may be noted without complicating factors. For such purposes

the eggs of Haminea are ideal.

Literature Cited

1918. BovARD, J. F., and Osterud, H. L. Partial list of Animals Yielding Embryological

JMaterial at the Puget Sound Biological Station. Pub. Puget Sound Biol. Sta.,Vol. 2,

No. 42.

1918. Leonard, Ruth E. Early Development of Haminea. Pub. Puget Sound Biol. Sta.,

Vol. 2, No. 34.

1922a. Richards, A. The Egg Laying Habit of Haminea Virescens (Sby). Proc. Okla.

Acad. Sci., Vol. H.

1922b. Richards, A. The Acceleration of the Rate of Cell Division. Biol. Bull., Vol. XLIII.

1901. Smallwood, Martin. The Centrosome in the ]\Iaturation and Fertilization of Bulla

Solitaria. Biol. Bull., Vol. 2.

1904a. Smallwood, Martin. Natural History of Haminea Solitaria (Say), .^mer. Nat.,

Vol. XXXVHI.

1904b. Smallwood, Martin. The Maturation, Fertilization, and Early Cleavage of

Haminea Solitaria (Say). Bull. ]\Ius. Comp. Zool., XLV.

DEPARTMENT OF METHODS, REVIEWS, ABSTRACTS AND

BRIEFER ARTICLES

NOTE ON THE OCCURRENCE OF THE MARINE CLADOCERAN

EVADNE TERGESTINA IN SOUTHERN CALIFORNIA

By

Helen E. Murphy

Dr. Chancey Juday reported the presence of the marine Cladoceran

Evadne tergestina Claus in Southern CaUfornia in 1907. He states: "This

Cladoceran was found in only eleven catches, eight of which were surface."

(Univ. of Calif. Publ. in Zool. vol. 3, no. 10, pp. 157-158.)

An enumeration of three series of catches of zooplankton collected

from the surface at 8 a.m. at the Scripps Institution, La Jolla, California,

enables me to amplify this brief record. A series extending from July

16, 1922 to May 1, 1923, contained specimens of this species from August

to late November, and again in April. The maximum production was in

April when there were many free swimming young, and females carrying

young. A second, though smaller maximum occurred in October and

November. Females carrying young predominated at that time. Two

smaller series, September 1916 to January 1917, and September 1919 to

January 1920, show adults present from September to January, with

immature forms in November.

METHODS OF CULTURING TUBIFICIDAE IN THE

LABORATORY

By

Eugene F. Powell

Tubificid worms are important in the laboratory because they are

forms whose Hfe history can easily be studied and because they are excellent

food for small aquatic animals such as fish, planarians and leeches. *Przi-

bram (1922) states that these worms may be kept for some time in tubs or

troughs of muck in which there is much plant detritus and over which

water is allowed to flow gently, the temperature being kept low and con-

stant.

* Hans Przibram-Wien : Das lebende Tiermaterial fur biologische Untersuchungen

(Auswahl, Beschaffung, Haltung unter verschiedenen Bedingungen, Markierung). Aber-

halden: Handbuch der biologischen Arbeitsmethoden. Abt. IX, Teil 1, Heft 2. 1922.

155

EUGENE F. POWELL 156

The writer started a culture October 9, 1922 for the purpose of study

and to provide food for some aquatic animals already mentioned. The

worms were obtained by two methods: (1) by washing out some of the

muck by the use of sieves and (2), by taking the material a^ found in a

river bed. The latter method resulted in the taking of many more worms.

Large glass dishes or pans were used in which the muck containing the

worms was placed to the thickness of about two and one half inches. For

a time the worms thrived but within a few weeks an appreciable decrease

in numbers was noted. Thinking that lack of food was the cause of the

decrease some boiled potato was added to the culture and within a few

days such good results were seen that a small amount of potato was

added about once in every two weeks thereafter. At present (April 30,

1923) the culture is still thriving and the worms are larger and greater in

numbers than when taken. It would seem that by the addition of boiled

potato or other similar food at frequent intervals a culture could be kept

going indefinitely.

A NEW METHOD FOR WHOLE MOUNTS

By

William K. Bowen

Michigan Biological Supply Co.

The destruction of microscopic preparations, due to careless focussing

of the microscope, is an occurrence with which biology instructors are only

too familiar. In the case of sections, it is often possible to dissolve off a

broken cover, and replace with another, but with whole mounts, the speci-

men is usually ruined. The writer has recently worked out a method,

applicable to a large number of whole mounts, which practically elim-

inates any possibility of injury to the specimen, however hard the

objective may be forced down upon the mount.

This method consists essentially in embedding the specimen in cel-

loidin, and in clearing and mounting the celloidin with the specimen within

it. The process is briefly as follows: Stain and dehydrate the specimens

just as if they were to be mounted in the customary manner. Place for

one to several hours in ether alcohol, the time required depending on the

size and density of the specimens. Replace the ether alcohol with thin

celloidin. Even large and relatively impermeable objects will be suf-

ficiently saturated in 24 hours. Transfer the celloidin and the specimens to

a perfectly fiat dish or suitable embedding tray. Orient the objects so

that when the mass is cut up, each specimen will be surrounded with a mass

of celloidin having an area of about half that of the cover glass to be

employed. Allow the celloidin to thicken by exposure to the air. When

the mass is hardened it should be as near as possible the same thickness as

WILLIAM K. BO WEN 157

the specimen. The amount of celloidin to use can be learned only by

experience. As soon as the mass has become sufficiently firm, complete the

hardening in chloroform vapor. Remove from the mould, and clear.

Finally cut the mass up into blocks, each containing a specimen and mount

in balsam. It is preferable to defer the cutting up of the mass until it is

cleared, as otherwise the small blocks will sometimes curl.

With this method the specimen is surrounded by a tough, resistant

matrix, and it is practically impossible to force an objective down on

such a mount with sufficient force to cause the least injury to the object.

Even if the cover is broken, the object is spared, and can easily and safely

be remounted. However, the celloidin acts as a cushion, and ordinarily

prevents the breaking of the cover unless the pressure upon it is very great.

This method has several other advantages. (1) The spec'men may

be transferred from the clearing fluid to the slide without any risk of

injury. Hydra, for instance, can be thus mounted without the breaking

off of buds or tentacles. (2) The use of rings, glass rods, or other cover-

glass supports is entirely eliminated, for the celloidin not only prevents

crushing of the specimen by pressure of the cover, but it also holds the

cover level. (3) The specimen does not have a tendency to drift out of

place when the cover is apphed, as often happens if the usual method is

used.

It is believed that this method will be found of considerable value in

the preparation of slides for class use. Although it is often more tedious

than the ordinary method of mounting, the insurance of the specimen

against injury more than justifies the extra trouble.

TRANSACTIONS

OF THE

American

Microscopical Society

Organized 1878 Incorporated 1891

PUBLISHED QUARTERLY

BY THE SOCIETY

EDITED BY THE SECRETARY

PAUL S. WELCH

ANN ARBOR, MICHIGAN

VOLUME XLII

Number Four

Entered as Second-class Matter August 13, 1918, at the Post-office at Menasha,

Wisconsin, under Act of March 3, 1879. Acceptance for mailing at the

special rate of postage provided for in Section 1103, of the

Act of October 3, 1917, authorized Oct. 21, 1918

GEORGE BANTA PUBLISHING COMPANY

MENASHA. WISCONSIN

1923

TABLE OF CONTENTS

For Volume XLII, Number 4, October, 1923

Descriptions of Two Specimens of Twinned Earthworms of Which One is Adult, with

two plates, by Frank Smith 159

A Revision of the Description of Diplocardia michaelseni Eisen, with two figures, by

Frank Smith 175

Recent Work on Marine Micro-Plankton at the La Jolla Biological Station, By W. E.

Allen 180

A Key to the Genera of Acanthocephala, with one figure, by H. J. Van Cleave 184

Department of Methods, Reviews, Abstracts, and Briefer Articles

The Pocket Microscope "Tami" 192

A New Pocket Microscope 193

A New Drawing Apparatus 194

List of Members Admitted Since the Last Published List 197

Index to Volume XLII 199

TRANSACTIONS

OF

American Microscopical Society

(Published in Quarterly Instalments)

Vol. XLII OCTOBER, 1923 No. 4

DESCRIPTIONS OF TWO SPECIMENS OF TWINNED

EARTHWORMS OF WHICH ONE IS ADULT*

By

Frank Smith

University of Illinois

The first earthworm specimen described in this paper differs in some

important characters from any others that have been seen by the writer,

and from any adult specimens of which he has thus far found descriptions.

It is unique among a rather extensive series of abnormal specimens which

have been acquired in the course of a study of the earthworm fauna of the

banks of a stream in the vicinity of Urbana, IlHnois. Several thousands

of earthworms have been collected, and more than fifty of them have

shown external signs of abnormalities among the reproductive organs,

but the specimen to be described is the only one which has shown evidence

of a double or twin type of structure involving the anterior somites.

That two individuals are represented is the obvious explanation for the

eight pairs of setae on each of about 40 anterior somites; for the four

oviducal and four spermiducal pores; and for the four distinct tubercula

pubertatis, instead of the normal number of two. In the anterior two

fifths of the length of the specimen this twin condition prevails, and then

a rather abrupt reduction in the diameter is accompanied by a reduction

of the setae to four pairs per somite; and the posterior three fifths of the

specimen has the appearance of that of a normal individual.

A desire to know something of the internal organization of such a

combination led to the preparation of transverse sections of the anterior

15 somites and of a few somites which include the posterior end of the

smaller individual.

Preliminary to a more detailed description, a general statement of the

relations of the two individuals represented and of some of the principal

organs will be useful. The species represented is Helodrilus tetraedrus

(Savigny). This is a highly variable species with several varieties or forms

recognized by the systematists, but the specimen belongs to the typical

form having spermiducal pores on the 13th somite, and anterior to the

*Contributions from the Zoological Laboratory of the University of Illinois, No. 227.

159

160 FRANK SMITH

oviducal pores. Apparently two individuals of unequal size and number

of somites are involved. The smaller one was fused throughout its length

with the anterior part of the larger one, and but one mouth and alimentary

tract functioned for both. The dorsal part of each individual, including the

dorsal vessel, maintained its integrity and apparently the dorsal surface

of the smaller specimen was next to the substratum and to that extent

served as the ventral surface for the aggregate; but, the specimen having

been killed before its peculiarities had been noted, nothing is positively

known of its ordinary habits of locomotion nor which side was ordinarily

next to the substratum. Since the length of the smaller individual was

less than one half that of the larger, the arrangement suggested would

result in having the ventral surface of the posterior half of the larger

specimen in its normal relation to the substratum. There are two ventral

nerve trunks, each with accompanying ventral and subneural blood

vessels; and a line connecting the nerve trunks, in transverse sections of the

specimen, would intersect at right angles a line connecting the two dorsal

vessels, but at a point somewhat nearer to the dorsal vessel of the smaller

individual. Each of these nerve trunks is apparently the result of the

fusion of components belonging to both individuals, and instead of being

in positions opposite to the dorsal vessels, they lie along what the casual

observer would naturally consider as the lateral walls of the composite

specimen. Figures 1, 2, 5, and 6 will aid in understanding these relations.

For convenience the larger individual will be designated as A and the

smaller one as B. Although each individual has its own dorsal parts

intact, the right and left sides of its ventral parts are separated from each

other and united with corresponding ventral parts of the other individual.

The right ventral half of A is fused with the left ventral half of B and

forms one of the two lateral surfaces of the composite specimen. Similarly

the left ventral half of A is fused with the right ventral half of B and

forms the other lateral surface. Corresponding relations prevail among

internal organs related to the ventral parts, such as the reproductive

organs and hearts. The relations are strikingly similar to those described

by Vejdovsky (1888-92) as existing in a juvenile specimen represented in

figure 14, of plate XIX, in his paper, and described as "A) Die Doppel-

missbildung, wo die Individuen mil den Bauchfldchen der ganzen Korper-

lange nach verwachsen sind."

In the following description it will be assumed that the use of the

expressions right, or left, side of the specimen refers to the side which

includes the corresponding one of the A individual. A reference to the

right side of the specimen will be to the one which includes the right side

of A and the left side of B.

In addition to the characteristics of the specimen due to the double or

twin structure, we find still further divergence from normal clfaracters due

TWO SPECIMENS OF TWINNED EARTHWORMS 161

to irregularities in metamerism. Both the compound metamere and the

spiral modification (Morgan 1895) are represented, and furthermore,

in some instances, there are wide discrepancies between the external and

internal indications of metamerism, such as intersegmental grooves, setae,

and septa. The extremely close relationship. between the individuals is

well illustrated by the manner in which they are affected by these meta-

meric irregularities. Wherever there is asymmetry among the organs and

parts of one individual there is corresponding asymmetry in the closely

associated parts of the other individual. The right side of A and the

intimately associated left side of B differ in the same manner from the

left side of A and the right side of B. Specific illustrations will appear

in the details of the following description.

External Characters

It will somewhat simplify the description to adopt a system for desig-

nating the setae which is commonly used by systematists. It is customary

to indicate the setae of either side by the use of the letters abed; the

ventralmost seta being designated by a, the next by b, the next by c, and

the dorsalmost one by d. The ventral pair includes a and b, and cd indi-

cates a dorsal pair. The dorsal interval between dorsal pairs is d — d,

and a — a is the interval between two ventral pairs. The description

will be further simplified by adopting the custom of using arable numerals

to indicate the different somites and of numerical symbols for the septa

and intersegmental grooves. The symbol 6/7 indicates either the septum

or intersegmental groove separating the sixth and seventh somites. In

the account of irregularities in metamerism, the term metamere will

commonly be used instead of somite which is its equivalent in oligochaete

terminology. This is done for the sake of conformity with the terms used

by Morgan in his article describing such irregularities.

The specimen was killed moderately well extended and is 91 mm in

length. The diameter varies from about IY2 n^^i in the genital region,

and about 3 mm in the clitellar region, to about V/^ m^a or less in the

posterior half. The number of somites in A is about 114, and in B is a

little over 40. Next posterior to intersegmental groove 7/8 there are two

grooves encircling the entire trunk, giving the appearance of two complete

somites to what is really a compound metamere or somite with two half

metameres on the right side of the specimen and but one half metamere on

the left side. That it is a compound metamere is shown by the presence

of two full sets of setae on the right side and but one on the left side, and

is also shown internally by the septal relations and the contained organs

(fig. 2). Posterior to this compound metamere follows a single somite

and then another compound metamere, as shown by the setae and internal

organs, but surrounded by two complete grooves. This latter compound

162 FRANK SMITH

metamere has the two half metameres on the left side and the single one

on the right side. Up to this point, the number of somites indicated by the

intersegmental grooves is one greater than the actual number, indicated

by setae and internal organization. The second compound metamere is

the beginning of a spiral modification, as shown by setae and by internal

organization. Superficially, as shown by the grooves, the spiral begins

with the groove 13/14, and extends posteriad five somites where normal

segmentation is resumed. A half metamere is intercalated on the left

side between somites 32 and 33 and a compound metamere with the two

half metameres on the right side follows tiext posterior to the somite which

includes 40 of the right side and 41 of the left. Somites posterior to 42

have normal numerical relations between their component half metameres.

In the specimen under consideration, the setal order for a somite,

beginning with the dorsal side of A, is d c b a (of A) ; a b c d d c b a (of B) ;

a b c d (of A); the aa setae being on the lateral surfaces, instead of on the

ventral surface as in normal specimens. The setae first appear, as usual,

on somite 2; four pairs or bundles in A, and but one in B. In 3, eight

pairs are present, as in each of the somites next following; but the setae

of B are very closely crowded. Eight is the usual number until somite

40 of the right side (41 of the left side) is reached, which has but six pairs.

41 of the right side is part of a compound metamere and has the dorsal

and ventral pairs of the right side of A, and only the ventral pair of B.

42 of the right side is continuous with 42 of the left side, constituting the

remainder of the compound metamere, and has six pairs. 43 has five pairs

of which but one belongs to B. All of the succeeding somites have each

but four pairs of setae, all belonging to A. The spacing of the setae in each

pair is about normal for the species, but the dorsal interval d — d of B is

only about one third as great as that of the same interval of A. This is

the approximate ratio in the genital region and posterior to it. In a few

anterior somites the discrepancy is still greater. In the fifth somite, d — d

of A is nearly five times greater than in B. In somite 10, the setae of the

ventral pairs are specially modified as genital setae. They are about twice

as long as ordinary setae, rather slender, and are associated with glands

which open into the setal apertures in the outer cellular layer in the body

wall. Similar modified setae with like relations and location ordinarily

occur in normal individuals. There is no apparent inequality in the size

of the setae of the two individuals.

The clitellum is slightly asymmetrical which is probably due to the

irregularities in metamerism. It includes ^ 22 — 27 on the right, and

23 — 3^ 28 on the left side of A. Four well defined tubercula pubertatis

are present, of which three are represented in figure 1. Those figured

include the two belonging to B and the one on the right side of A. Two

pairs of spermiducal pores are borne on i)rominent elevations of the body

TWO SPECIMENS OE TWINNED EARTHWORMS 163

wall on 13, and two pairs of inconspicuous oviducal pores are on 14. These

pores have the same general relations to the setal bundles of the individuals

A and B that one ordinarily finds in normal worms of this species, but the

peculiar relations of A and.B lead to the distribution of pores, from each

of the two individuals, along each of the two lateral areas of the composite

specimen. There is one spermathecal pore on the right side of A, in the

groove 9/10; and two on the left side, in the grooves 9/10 and 10/11.

There are none in B.

Internal Characters

There is no internal separation between the body cavities of the two

individuals. The space between two consecutive septa is continuous

between the two individuals to the same extent that it is between the two

sides of either one of them. The coelomic spaces of B are much more

restricted than are those of A and are nearly filled by the contained organs.

Mention has already been made of certain internal irregularities. Follow-

ing the seven anterior somites which are normal, a compound metamere

results in having 8 and 9 of the right side of the specimen with a dividing

septum, opposite to and in communication with 8 of the left side, which

lacks the dividing septum. This is followed by 10 of the right side opposite

9 of the left side. Then follows a compound metamere or somite in which

10 and 11 of the left side are opposite 11 of the right side. Due to the

introduction of a spiral modification in connection with this second com-

pound metamere and extending posteriad for several somites, the spirally

wound septum which begins anteriorly with the septum between 10 and

11 of the left side of the compound metamere allows free communication

between the cavities of the constituent half metameres with each other,

and with those following in the spiral (fig. 2). The general outcome of

these complications is the normal location of most organs, if septa and

setae are adopted as criteria for somite limits, as they are in this paper.

As previously stated, one alimentary tract functions for both individ-

uals. It differs in structure from that of a normal worm and apparently

includes at least the dorsal portions of the alimentary tracts of A and B.

This is apparent from the structure and from the relations to the dorsal

vessels. The normal condition in the fourth and fifth somites, in which

there is a pouch formed by an evagination of the dorsal wall of the alimen-

tary tract surrounded by an extensive mass of gland and muscle tissue,

is replaced in the twinned specimen by such a pouch on each of the two

sides of the pharyngeal lumen which correspond to the dorsal walls in both

A and B. The one belonging to B is a little smaller than the other, but

none the less typically developed. In normal specimens, a pair of evagin-

ated pouches are formed in 10 from the latero-dorsal walls of the esophagus

and are a part of the calciferous gland which is formed in the walls in 10

164 PRANK SMITH

and a few following somites. Two normally formed pouches are present

in A and are in the 10th somite, but the asymmetry due to irregularities in

metamerism result in the pouch of the right side of A being formed anterior

to the one on the left side. Instead of evaginated pouches, the wall of the

alimentary tract in 10 of B is invaginated at the locations where pouches

are normally formed. Instead of the normal typhlosole on the dorsal side,

a typhlosole is developed on each of the two opposite walls of the intestine

which are interpreted as being the dorsal parts of the two intestines of A

and B united into a single tube. The typhlosole of B is not represented

posteriad of the anterior part of the 44th somite.

The circulatory system has very interesting deviations from normal

relations. A and B each has a dorsal Vessel, and accompanying each

nerve cord is a ventral and a subneural vessel. Each dorsal vessel with its

branches presumably has its origin wholly from one individual, while each

ventral and subneural vessel has probably had a double origin, as have

also the vessels connected with them. The hearts, as in normal specimens,

are paired in 7 — 11 of each individual and connect the dorsals with the

ventral vessels (fig. 5). Hearts of the right side of A connect with the

ventral vessel of the corresponding lateral side of the specimen, which

also receives the hearts of the left side of B. Similarly the hearts of the

left side of A connect with the ventral vessel of the left side of the specimen,

as do also the hearts of the right side of B. Irregularity in metamerism

has led to asymmetry in the relations of hearts on opposite sides of each

of the dorsal vessels, but has not disturbed the paired relations between

the hearts on the opposite sides of either of the ventral vessels. The

large paired latero-longitudinal vessels which normally join the dorsal

vessel in 12 are present in that somite in both A and B, one pair in each.

The dorsal vessel of B extends to the anterior part of 44. The posterior

end communicates with the vascular plexus in the intestinal wall through

several large connecting vessels. The subneural vessels unite in 46, and

the ventral vessels in the next following somite.

The nephridia are paired in most somites of each of the individual

components of the specimen, and the locations of the nephridial funnels

and external pores have normal relations in each. Posteriorly, the ne-

phridia of B are not represented posterior to 45, and somites 43 to 45 have

each but one nephridium, with pores very near the median line.

The study of the nervous system has been limited to an examination

of the relations of the niost important ganglia and ventral nerve cords.

The supra-esophageal ganglion or "brain" of A has the normal position

and gives rise to a pair of circumesophageal trunks of normal size, of

which the one on the right side is continuous with one half of the ventral

cord in the right side of the specimen, and the one on the left side is con-

tinuous with one half of the other ventral cord. Similarlv the "brain"

TWO SPECIMENS OF TWINNED EARTHWORMS 165

of B, which is slightly smaller than that of A, is normally situated with

reference to the pharyngeal wall of B and is connected by circumesophageal

trunks with each of the ventral nerve cords (fig. 6). The trunk on the

left side of B joins the one from the right side of A, each being continuous

with that side of the ventral nerve cord which is related to the individual

to which the trunk belongs. Similarly the circumesophageal trunk on

the right side of B joins the one on the left side of A and each is continuous

with a corresponding half of the ventral nerve cord on the left side of the

specimen. Near the posterior part of B, the two ventral nerve cords

approach each other, and in 44 and 45 there is a considerable decrease in

the size of the half of each of them which belongs to the B individual. The

first actual union of the two cords is in the anterior part of 46. An examin-

ation of sections of the cords, slightly anterior to the point of union dis-

closed the following facts concerning the relations of the three "giant

fibres" in each of the cords anterior to the union, to those of the single

cord posterior to the union. The outer giant fibre in each cord which

was related to B disappeared; the middle and largest fibres of the two cords

united into one; and the outer fibres in each cord which belonged to A

was continued into the single cord, posterior to the union (figures 3 and 4).

The location of the principal reproductive organs in normal worms of

the species under consideration is as follows: one pair each of testes and

of spermiducal funnels in each of somites 10 and 11; one pair each of

ovaries and of oviducal funnels in 13; one pair of sperm sacs in each of

somites 9, (occasionally 10), 11, and 12; and one pair of spermathecae

opening dorsally in close relation to each of the septa 9/10 and 10/11.

The various reproductive organs of each individual of the twinned speci-

men are of normal size and location, with the exception of the spermathecae

which are unrepresented in B and of which A has but three. There is

no spermatheca in the right side of A, related to the septum and interseg-

mental groove 10/11. The various organs of the right side of A are dis-

tributed along one side of the nerve cord of that side of the specimen,

and on the other side of the nerve cord are arranged the corresponding

organs of the left side of B. The organs on the two sides of the nerve cord

have a paired relation, but the constituent members of any pair belong

to the different individuals A and B. Similarly the organs of the left

side of A are associated with those of the right side of B, along the nerve

trunk of the other side of the specimen. The irregularities in metamerism

involve a correlated asymmetry among the organs on the two sides of

either individual, but have not resulted in asymmetry in the arrangement

of the organs on the two sides of the same nerve trunk. To illustrate:

The testis and spermiducal funnel of the right side of somite 11 in A are

opposite those belonging to the left side of somite 11 in B, but they are

also opposite those of. the left side of somite 10 in A.

166 FRANK SMITH

The transition from the twinned condition to that of a single individual

involves a somewhat gradual but continuous reduction in the body wall

of B. This reduction begins with the disappearance of the mid-dorsal

part of the wall and extends farther and farther laterad as one follows the

sections posteriorly. In somite 40 (of the right side of the specimen),

the dorsal pairs of setae of B and the part of the wall indicated by d — d

have disappeared. In 43, the parts of the wall indicated by b — c have

almost disappeared and the two longitudinal areas which include the

ventral pairs of setae are closely approximated, and only one of these

pairs develops in this somite. A gradual reduction of the remaining

part of the wall a — a in the next two somites is followed by its complete

disappearance in the anterior part of 46 where the union of the ventral

nerve cords and accompanying trunks of the vascular system complete

the transition from the twinned part of the specimen into that of the

single individual.

General

In the foregoing description it has been assumed that the specimen

studied includes two individuals united along their ventral surfaces.

A discussion of this assumption involves references to a few publications

dealing with the same general subject. Vejdovsky (1888-1892) has

contributed an extended account of the results of a study of twinning in

several species of earthworms and an important discussion of the general

subject. More recently Weber (1917), Welch (1921), and Newman (1923)

have published in the same general field. Since some of these papers

have bibliographic lists, it is unnecessary to introduce such lists in this

paper. One species of earthworms which is very abundant in Europe and

North America has been studied by several investigators and has been

shown to produce, during early stages, large numbers of twinned specimens.

This species has received various names in the literature on the subject,

due to nomenclatural changes. Lumhrictis Irapezoides, AUoJobophora

trapezoides, and Helodrilus caliginosus Irapezoides are simply different

names for the same species, as has already been noted by Welch. Vejdov-

sky found very many twinned specimens among the embryos and young

obtained from cocoons of this species. He also found great variety in the

mode or type of union which the twinned specimens exhibited. Of these

various types, he gives chief attention to three. "Demnach werde ich im

Nachfolgenden nur die charaktcristischen Doppelembryonen anfuhrcn,

solche namlich, deren Verwachsungsflachen man ganz sicher ermitteln

kann." "A) Die Dappelmissbildung, wo die Jndividucu mil den Bauch-

jldchen der ganzcn Korpcrliinge nach vcrwachsen sind. Ich fand mehreremals

diese interessante Missbildung, immer je eins im Cocon. Die grosste

war 8 mm. lang und ich habe sic auf dcr Taf. XIX, Fig. 14, nach dem

Leben abgebildel." This figure is reproduced in figure 11 of this paper.

TWO SPECIMENS OF TWINNED EARTHWORMS 167

It has an appearance, at the posterior end, of dissimilarity in the size of

the two individuals involved, and is very suggestive of what may have

been the appearance of the specimen described in this paper, when it was

at a similar stage of development. Vejdovsky has figures of six transverse

sections made from different parts of his specimen, which indicate the

same plan of organization as that described in this paper. One alimentary

tract representing the dorsal parts of the alimentary tracts of two individ-

uals; each individual with its own brain, dorsal vessel, nephridia, and

setae in normal relations. Two ventral nerve trunks, double in nature.

"Es ist ersichtlich, dass jedes Bauchstrangsganglion einer Hiilfte des

einen und einer Halfte des anderen Individuums seinen Ursprung ver-

dankt."

In a more recent paper (Weber 1917), we have the results of an examin-

ation of the contents of 184 cocoons of North America representatives of

the same species. Specimens were found representing various stages of

development, from early cleavage to individuals with more than 100

somites. 35 specimens were twinned, and of these, twelve were sufficiently

developed to have 60 or more somites in each individual. In six of the

twelve, there was a dorsal union of the component individuals, but in no

one of them did this union involve more than the anterior five somites.

In none of the twelve specimens was there a union along the ventral

surfaces and the author asserts the probability that the specimen des-

cribed at length by Vejdovsky, as an illustration of a union along the

ventral surfaces, was really one in which the union was along the dorsal

surfaces. She admits that the evidence for her view is not conclusive, since

in Vejdovsky's specimen the individuals were united along their entire

length; but the similarity of the relations between organs, as represented

in Vejdovsky's figures, to those shown by sections of her specimens in

which she felt sure of dorsal surface union led her to assume that Vejdov-

sky's interpretation was erroneous. She claims that the pharyngeal rela-

tions as shown in Vejdovsky's figures support her views. The writer sees

nothing in these figures that is contrary to what might be expected in the

event of a union along ventral surfaces. The striking similarity of the

adult specimen described in this paper, to Vejdovsky's specimen make

the same type of union for both highly probable. If the posterior three-

fifths of the writer's specimen is really a part of but one of the individuals

represented in the anterior twinned part, then the union must have been

along the ventral surfaces. The claim that the union is along the dorsal

surfaces involves the assumption that the posterior three-fifths of the

specimen is made up of two parts, lying on either side of the sagittal plane,

which have originated in two separate individuals, and that for some

mysterious reason the other half of each has completely disappeared.

There seems to be no basis for any such assumption and the writer believes

168 FRANK SMITH

that his specimen belongs to Vejdovsky's "A" type of twinned earthworms.

The second type of union, as treated by Vejdovsky, is: "B) Die Doppel-

missbildung, deren Individnen Icings der Riickenseiten verwachsen, fand ich

nur in einem einzigen Falle." This type of union which was represented

by but a single specimen among those studied by Vejdovsky was found in

six of the twelve specimens described by Weber. The third type is: ''C)

Die polar verwachsenen Doppelmissbildungen sind bei Allolobophora trape-

zoides die haufigsten, aber auch die Modificationen dieser Verwachsung

sind sehr zahlreich." This type of union was found in three specimens of

the twelve older ones in the Weber material.

The species in which Kleinenberg, Vejdovsky, and Weber found a

twinned condition among the young taken from cocoons to be of common

occurrence is one of the most widely distributed and abundantly repre-

sented of any of the Lumbricidae of Europe and North America. Appar-

ently very few, if any, survive from among the specimens in which the

union involves any considerable number of anterior somites. No mention

of such specimens in adult stages has been found by the writer, and no

such specimens have been found by him or his assistants during an exam-

ination of several thousands of worms of this species. The specimen

described in this paper belongs to the same genus, but to a species of which

the early developmental stages have not been studied and reported, as

far as is known to the writer. Apparently the twinned condition afforded

no obstacle to success in the struggle for existence of this particular speci-

men, since it is one of the largest found among several hundred individuals

of this species collected in this vicinity. Whether twinning occurs fre-

quently, or rarely, among the embryos of this particular species is yet to be

determined.

Welch in his recent paper on bifurcation of the embryos of Tubifex

offers an explanation for the rarity of adult twinned specimens, even when

the twinned condition is common among embryos. It is due to the almost

insurmountable difficulties which twinned specimens encounter in their

efforts to escape from the cocoons which are provided with openings

barely large enough to allow the exit of normal specimens. He found also

none surviving for any great length of time among the few twinned speci-

mens that were released from the cocoons. For this latter fact no explana-

tion is obvious.

The last few chapters of Newman's Physiology of Twinning (1923)

include a discussion of symmetry reversal and mirror imaging. It is a

subject which has received attention from a number of workers and one

of which the explanation involves diversity of opinion. Normal earthworms

are so thoroughly bilaterally symmetrical that they offer little opportunity

for a study of symmetry reversal. The adult specimen described in this

paper with its marked asymmetry between the two sides of either individual

TWO SPECIMENS OF TWINNED EARTHWORMS 169

furnishes an excellent illustration of symmetry reversal, and one which

may be of interest to those striving to arrive at explanations of such

phenomena. In this case the reversal involves organs which are in ab-

normal relations, and is obvious because of this abnormality. Presumably

the special features of the abnormal relations are peculiar to the one

generation and do not extend back through a long series of ancestors.

Twinning in Sparganophilus Eiseni

Welch in referring to the literature on bifurcation among Oligochaeta,

makes this statement concerning embryonic bifurcation: "All recorded

instances fall within the Lumbricidae." In his paper he has shown that

it also occurs within the family Tubificidae. A recent paper (Hague, 1923)

has made known that this phenomenon is found occasionally in Spargano-

philus eiseni which belongs to the family Glossoscolecidae, or, if we follow

Michaelsen (1921), to a more restricted family Sparganophilidae. Four

juvenile specimens of this species which were taken from cocoons are

bifurcate posteriorly and exhibit a dorsal type of union, anteriorly, of

7 — 25 somites. One of these specimens which had been collected at Douglas

Lake in Cheboygan County, Michigan, was presented by Dr. Hague to

the writer and from it figure 10 was prepared, showing some of the char-

acters recognizable in it as an unstained specimen in cedar oil. The

anterior 15 somites of the two individuals were united and the two separate

posterior parts each contained about 25 well defined somites and a terminal

growing region. Transverse sections of the anterior twinned part show

the presence in each component individual of a mouth and alimentary

tract in the first five somites. In the next few somites these are united

into a single esophageal tube. Posterior to the 14th somite the alimentary

tracts are again separate and extend into the two posterior trunks. In

close contact with each other and between the two alimentary tracts,

lie the supraesophageal ganglia of the two component individuals (fig.

7, 8, and 9, br.) and each is connected by circumesophageal trunks with

the corresponding ventral subesophageal ganglion. There is no separa-

tion between the coelomic cavities of the two individuals in the anterior

twinned part. Nothing has been found of a well defined circulatory

system, but traces of what appear to be the beginnings of dorsal vessels

are found on either side of the alimentary canal and about equidistant from

the two ventral nerve cords.

Summary

A description is given of the anatomy of a twinned specimen of Helod-

rilus tetraedrus which had attained sexual maturity.

The general organization is similar to what might be expected if one

should succeed in splitting the anterior 45 somites of a normal individual

170 FRANK SMITH

in a sagittal plane from the midventral surface as far as the middle of the

lumen of the alimentary tract; then in spreading the cut surfaces apart

and inserting a specimen similarly treated which had a corresponding

length and number of somites (placing the cut surfaces in contact), thus

producing a specimen with a double number of setae, reproductive organs,

nerve and vascular trunks and nephridia, but with a single alimentary

tract and mouth.

The double or twinned condition prevails in the anterior 45 somites and

is followed by about 70 somites of the type of organization found in the

posterior part of a normal individual.

Certain irregularities in metamerism with consequent asymmetry in

some of the reproductive and associated organs are described.

A brief description is given of a juvenile twinned specimen of Spar-

ganophilus eiseni, in which there is a dorsal type of union of the anterior

15 somites of the two individuals, followed by separate posterior parts of

about 25 somites each.

Literature Cited

Hague, Florence S.

1923. Studies on Sparganophilus eiseni Smith. Trans. Am. Micr. Soc, 42: 1-38, 4 pi.

Kleinenberg, N.

1879. The Development of the Earth-worm, Lumbricus trapezoides Duges. Quart.

Journ. Micr. Sci. (N.S.), 19: 206-244, 3 pi.

MlCHAELSEN, W.

1921. Zur Stammgeschichte und Systematik der Oligochaten, insbesondere der Lum-

briculidae. Archiv. Naturg. 86: 130-142.

Morgan, T. H.

1895. A Study of Metamerism. Quart. Journ. TSIicr. Sci. (N.S.), 37: 395-476, 4 pi.

Newman, H. H.

1923. The Physiology of Twinning. Chicago.

Vejdovsky, Fr.

1888-92. Entwickelungs geschichtliche Untersuchungen, Prag.

Weber, Roxie A.

1917. Observations on the Structure of Double Monsters in the Earthworm. Biol.

BuU., ii: 339-354, 3 pi.

Welch, Paul S.

1921. Bifurcation in the Embryos of Tubife.x. Biol. Bull., 41: 188-202.

172 FRANK SMITH

Plate 1

Explanation of figures

1. Showing the dorsal part of B, in the posterior half of the twinned part of the specinien.

The lower part of the figure and a few somites at the right are from A, as seen from its

ventral side. CI, clitellum; t p B and t p A, tubercula pubertatis of B and A, respectively.

20 30, and 40, numbers of somites on left side of specimen.

2. Diagram showing the relations of parts and organs in somites containing the re-

productive bodies, represented as though the body wall had been split along the median dorsal

surface of A and then spread out and viewed from the inner surface, a, b, c, and d, setae;

d V, dorsal vessel; n c, nerve cord; A(l) and A(r), left and right halves, respectively, of A;

7-14, numbers of somites; 9 , oviducal pore; cT, spermiducal pore; o, ovary; s, testis or sper-

mary; o f, oviducal funnel; s f, spermiducal funnel; s s, sperm sac; s t, spermatheca; sep,

septum.

3. From transverse section near posterior end of B, showing union of the two ventral

nerve cords and near approach of the two ventral vessels and the two subneural vessels,

m f median giant nerve fibre; n c, nerve cord; s v, subneural vessel; v v, ventral vessel.

4. From a section slightly posterior to one shown in figure 3, showing closer approach

of parts. Same abbreviations as in figure 3.

5. Diagram showing relations of "hearts" and longitudinal blood vessels; d v, dorsal

vessel; h, heart; n c, nerve cord; v v, ventral vessel.

6. Diagram showing relations of brain, circumesophageal nerve trunk, and subesophageal

ganglion at anterior end of ventral nerve cord, br, brain; c t, circumesophageal trunk;

s g, subesophageal ganglion.

Plate 2

Explanation of figures

7 8 and 9. Oblique sections through anterior somites of specimen figured in 10.

Obliquity such that the part shown on the right side of each figure is approximately two

somites posterior to that shown on the opposite side, c d, fragments of setae of dorsal

pair.

7. From section through mouth opening on one side, and setae of third somite on opposite

side, a t, alimentary tract; br, one side of brain; m, mouth; c d, dorsal setae of third somite.

8. From section through region about one somite posterior to that shown in figure 7,

b r brain; s g, subesophageal ganglion; c d, dorsal setae of fourth somite.

9. From section through region about two somites posterior to that shown in figure 7.

a t, alimentary tract; b r, brain; c t, circumesophageal trunk; s g, subesophageal ganglion;

c d (at left side), dorsal setae of second somite; c d (at right side), dorsal setae of fifth somite.

10. From anterior part of twinned juvenile specimen of Sparganophilus eiscni, with

dorsal type of union, b r, brain; m, mouth; n c, nerve cord; p, prostomium (combined).

11. From figure 14, plate XIX, Vejdovsky (1888-1892). Showing twinned juvenile

earthworm with ventral type of union.

t p B

±hLLLLL:jJJj^^^

I p B t p A

Plate I

Plate II

A REVISION OF THE DESCRIPTION OF

DIPLOCARDIA MICHAELSENI EISEN*

By

Frank Smith

University of Illinois

Eisen's preliminary description of this species appeared in 1899 and a

more extended account in 1900. Each of these papers contained a review

of the various species of Diplocardia then known, and a suggestion of

the probable existence of a still greater number of unknown species. The

description of D. michaelseni was based on material from Raleigh, N. C,

collected by Messrs. Brimley, and Eisen states that he had a dozen speci-

mens. In 1899, the year in which the preliminary description was pub-

lished, Dr. Eisen sent to the writer several specimens of earthworms of

different species, one of which was accompanied by a label which reads

"Diplocardia michaelseni Eisen. Raleigh, N. C. April, 1898. Eisen det."

In the absence of any evidence to the contrary it seems natural to assume

that this may be one of the dozen specimens which Eisen mentions. In the

preparation of a paper dealing with new and known species of Diplocardia,

the writer, being desirous of certain data concerning Eisen's species which

were not supplied by his papers, has prepared and studied sections of some

of the specimens which Dr. Eisen had sent him. A result of this examina-

tion has been the discovery of a few discrepancies of some importance

between the description of D. michaelseni and the specimen supplied,

which will receive attention. Eisen states that only one of his specimens

of this species was sectioned. The writer has studied sagittal sections of

one half of the anterior 23 somites of his specimen.

It will simplify the comparison between Eisen's description of the

species and the results of this examination if we first consider characters

in which there is agreement. We have the following: "Prostomium

divides somite 1 completely." "Setae all ventral; a — a = 3a — b; a — a

about one third larger than b — c." Eisen makes no mention of the distance

c — d which in the writer's specimen is about IJ^ times a — b, and a — b is

slightly less than described by Eisen. Clitellum on 13 — 17 and encroaches

somewhat on the dorsal part of 18, and is but slightly developed ventrally

on 13. A "spermathecal genital zone" includes pairs of large rosette-like

papillae on the ventral side of 8 and 9. Setae a and b of these two somites

are highly modified and sculptured (Eisen, 1899, fig. 2). A "post-clitellar

genital zone" includes a median ventral depression on somites 18 — 20,

*Contributions from. the Zoological Laboratory of the University of Illinois, No. 228.

175

176 FRANK SMITH

connected anteriorly with two deep pits in the posterior margin of 17.

Somites 21-23 have on the ventral side two pear-shaped swellings. Only

the anterior one is well defined in the writer's specimen. Grooves connect-

ing the prostate gland pores of either side are approximately straight.

These pores are paired on 18 and 20 and the spermiducal pores on 19.

In the writer's specimen, the latter pores are near the anterior margin of

the somite. Oviducal pores on 14 are near the anterior margin and very

close to the mid-ventral line and to each other. The spermathecal pores

have relations which distinguish the species from the other known species

of the genus. There is one pair on 8, slightly anterior to the ventral pairs

of setae and each about equally distant from a and b of the same side. A

second pair is present on 9, but each opens near the posterior margin of

the somite, nearly in seta line a.

Septa 7/8 and 8/9 are more strongly thickened than others. Spermaries,

spermiducal funnels, ovaries, and oviducal funnels have the usual number,

positions, and relations. There are paired sperm sacs in 9 and 12. Two

pairs of prostate glands occupy a good deal of the available space in somites

17 or 18 to 21 inclusive. The muscular ducts are long and contorted and

open on 18 and 20. The glandular parts have a peculiar structure described

in detail by Eisen. The lumea of each is very inconspicuous, without

definite epithelial lining, and of no greater diameter than that of the duct.

Numerous branches of the lumen are formed and each is surrounded by a

single layer of gland cells. Spermathecae are paired in 8 and 9, and have

long contorted ducts, each of which has a long diverticulum, directed

anteriorly, and a definite sac-like expansion which has a length greater

than the diameter of the worm, and a strong constriction about midway

of its length. Bundles of glands, developed in the body wall, extend

parallel with the long axis of the body and open in close relation with the

apertures of the modified spermathecal setae. The development of these

glands is largely responsible for the papillae on the ventral surfaces of

8 and 9.

We have now to consider several features in which the specimen re-

ceived from Dr. Eisen differs from the description of D. michaelscni. In

the description we find: "Size 45 mm. by 2 mm., hardly tapering poster-

iorly. Somites 63." The writer's specimen is about 65 mm. by \Yi mm.

and has 116 somites. Presumably the type specimen on which the de-

scription was based had lost a number of somites from the posterior end.

In his first paper Eisen says: "Penial setae present at spermiducal pore"

and makes no mention of such setae at the prostate pores. In his second

paper the "Definition" which is chiefly a repetition of the matter in the pre-

liminary paper has the same reference to penial setae, while in the more ex-

tended description we find no reference to the setae at the spermiducal

pore, but a statement that the prostates "open as usual near sacs with

DESCRIPTION OF DIPLOCARDIA MICHAELSENI EISEN 177

penial setae, but I am unable to say whether these are sculptured or

smooth." The writer has found no setae at the spermiducal pores in his

specimen. The penial setae at the prostate pores are very inconspicuous

and but little could be learned of their characters in the relatively thick

sections containing them. As indicated above, there is general agreement

in the positions of the various reproductive organs.

Eisen states that "The enlargement of the septa is prmcipally dorsal,"

referring especially to 7/8 and 8/9. His figures indicate a specimen nearly

straight and not strongly contracted anteriorly. Figure 1 of this paper

shows the anterior part of the writer's specimen strongly contracted an-

teriorly and curved ventrally which may easily account for the fact that

the septa 7/8 and 8/9 are thickest ventrally. The same figure shows clearly

Fig. 1. Combination of parts of several sections with the aid of a camera lucida, show-

ing relations of the body wall, septa, and alimentary tract, g, gizzard; s s, distal part of setal

sac; st po, spermathecal pore; 5/6, septum between somites 5 and 6.

that the gizzards are in somites 5 and 6, and not in 6 and 7, as stated in

the description. Eisen makes special reference to a glandular crop in 14

and 15 and states: "As far as we know it also differs in the possession of a

glandular crop in two of the clitellar somites, similar to the one described

m Pontodrilus michaelseni." Eisen overlooked the similarity of this organ

to the one in the same situation in Dlplocardia eiseni Michaelsen which had

been described in detail by Michaelsen in 1894, and which is even some-

what more highly differentiated, and has more numerous and strongly

developed folds. Figure 2 shows something of the general character of this

organ in D. michaelseni, as represented in the writer's specimen. Its posi-

tion v/as somewhat oblique and sections that are nearly median in 15 are

more nearly tangential in the part lying in 14. There is much evidence in

support of Michaelsen's supposition that this organ is of the nature of a

calciferous gland.

In the description, Eisen states: "I find muscular connecting vessels

or hearts in 10 and 11 only. There is no supra-intestinal dilatation cf

the dorsal vessel, as in some other species. The dorsal vessel appears

to be single." In the writer's specimen, there is an additional pair of

178

FRANK SMITH

hearts in the twelfth somite which are fully as large as those of 10 and

11. The part sectioned did not extend quite far enough to permit a positive

statement, but the indications are that the hearts of 11 and 12, at least,

are dorso-intestinal hearts, by which is meant that they have their larger

dorsal opening into a supra-intestinal trunk, and a smaller connecting

vessel opening into the dorsal vessel. The supra-intestinal trunk has its

origin and termination in the vascular plexus of the wall of the alimentary

tract and is not a part of the dorsal vessel as implied in Eisen's statement.

In the description, it is stated that sperm sacs are present in 9, 10, and 12;

those of 10 being postseptal. This would imply a communication of

those in 10 through the septum into 9 in which there are no spermaries

nor sperm ducts. In the writer's specimen, there are no well defined

sperm sacs in 10 and there is but little difference in appearance between

the masses of sperm cells in 10 and 11.

Fig. 2. From a sagittal section through the alimentar>' tract in somites 14 and 15.

Section nearly median in posterior part of caliciferous gland and more nearly tangential in

anterior part. 13/14 and 14/15, septa between 13 and 14, and between 14 and 15.

In spite of the few discrepancies between Eisen's description and the

details of structure exhibited by the specimen, the writer is convinced that

the specimen really belongs to the species to which it was assigned by Eisen,

and that the discrepancies are due in part to oversight and errors of

observation in the preparation of the original description, and perhaps in

part to individual differences in specimens. Unfortunately the destruction

of Eisen's collections at the time of the San Francisco earthquake has

presumably made another examination of the type specimen impossible.

The septa related to the gizzards are quite thin and difficult to follow

in sections, and, if not considered, the gizzards are very likely to be allotted

to somites posterior to the ones to which they actually belong, since they

are commonly found shoved posteriad and lying under divisions of the

body wall which are a somite or two posterior to the ones to which they

are actually related. This is illustrated in figure 1 which represents the

DESCRIPTION OF DIPLOCARDIA MICHAELSENI EISEN 179

septa as somewhat more distinct than they actually are. This apparent

dislocation posteriad is common in Diplocardia. Such a condition is

perhaps related to the frequent eversion of the buccal cavity in normal

feeding activities, which must involve a good deal of flexibility and freedom

of movement in the anterior parts of the alimentary tract. It is difficult

to account for the statement that hearts were found in 10 and 11 only,

since no Diplocardia species or specimens are known to the writer, in

which the hearts are not present at least as far posteriad as the twelfth

somite.

The following characterization of the species includes the few changes

made necessary and is believed by the writer to be more accurate than the

original description.

Diplocardia michaelseni Eisen

Length, 45—65 mm. Somites, 63—116. Clitellum, 13— ^il^, but little

developed on ventral side of 13, cingulum. Papillae on 8 and 9 in close

relation to ventral setae and to glands developed in the body wall; one

or two pear-shaped swellings on ventral side of 18 — 20. Spermiducal

pores, paired on 19; prostate gland pores, paired on 18 and 20, those of

either side connected by nearly straight longitudinal grooves. Oviducal

pores, paired near anterior margin of 14 and near mid-ventral line. Sper-

mathecal pores, paired on 8, slightly anterior to ventral setae; and paired

on 9, but near posterior margin of that somite and nearly in seta line a.

aa:ab:bc:cd = 6:2:4:3. Ventral setae, of 8 and 9, highly modified as

genital setae; none on 19. Septa, 7/8 and 8/9 most heavily thickened.

Gizzards, in 5 and 6. Calciferous gland, with conspicuous longitudinal

folds, in 14 and 15. Dorsal vessel, double. Hearts, paired in 10 — 12.

Spermaries and spermiducal funnels, paired in 10 and 11. Sperm sacs,

paired in 9 and 12. Prostate glands, two pairs, 17 — 21. Ovaries and

oviducal funnels, paired in 13. Spermathecae, paired in 8 and 9; each

with one long diverticulum.

Literature Cited

Eisen, G.

1899. Notes on North American Earthworms of the Genus Diplocardia. Zool. Bull.,

2: 161-172.

1900. Researches in American Oligochaeta, with Especial Reference to those of the

Pacific Coast and Adjacent Islands. Proc. Cal. Acad. Sci., 3d Ser., 2: 85-276.

MiCHAELSEN, W.

1894. Die Regenwurm-Fauna von Florida und Georgia. Zool. Jahrb. Syst., 8: 177-194.

RECENT WORK ON MARINE MICRO-PLANKTON AT THE

LA JOLLA BIOLOGICAL STATION

By

W. E. Allen

Scripps Institulion, La Jolla, Calif.

In the earlier years of the Scripps Institution for Biological Research

and of the San Diego Marine Biological Association which preceded it,

all efforts at investigations were necessarily devoted to surveying the

field in quest of most promising avenues of research and to laying a founda-

tion of taxonomy, general morphology, general habitat range and general

distribution studies. In 1919 it was decided that sufficient progress had

been made in basic studies and in physical oceanography to warrant an

effort at more intensive work on diatoms and dinoflagellates, in addition

to some already in progress with the copepods and chaetognaths.

Local experience as well as authoritative literature had shown that

ordinary methods of collecting microplankton required a great deal of

skill, expensive equipment, considerable time and space and a number of

favorable conditions for operation. Furthermore, it was well known that

such methods could not yield very accurate results in estimating the

plankton population of the area sampled.

After special experiment the most satisfactory method was found to be

that of dipping up water in some kind of container and pouring it in

measured quantity through a small silk net or other filtration device.

Number twenty-five bolting silk was adopted as the standard filtering

material for diatoms and for other forms of similar size. For diatoms

twenty-five liters was found to be satisfactory as a standard quantity of

water. On account of accessibility the surface was arbitrarily selected

as the standard level and fixed inshore stations (such as piers) as standard

stations for collecting microplankton.

As a result of such methods and plans of operation the range and

continuity of collecting has been greatly extended. An unskilled workman

of careful habit can make just as good standard collections as can an

expert. Initial equipment for standard work costs only a small fraction

of the former ordinary cpcpcnditure in starling plankton work and the

operating expense is reduced far more since frequent replacement is no

longer required. The saving of time and space in making standard collec-

tions is also very great. In addition we have convincingly shown that

valuable series of collections can be taken under conditions formerly

prohibitive. It is also evident that statistical treatment of material

180

RECENT WORK ON MARINE MICRO-PLANKTON 181

obtained by standard methods yields estimates of population which are

more accurate than it is physically possible for the sampling to be. And,

of course, the continuity of series possible by such standard methods

strongly augments the value of the statistical records.

At our own pier twenty-five liter collections have been made daily

for a little more than four years, at Pt. Hueneme (except Sundays and

holidays) for nearly four years, and similarly at Oceanside for nearly three

years. These have been supplemented by fragmentary and short series

at Redondo, Santa Barbara and Pacific Grove. Such continuous series

could not have been obtained by us from even these few localities by any

other method.

At no time have we intended to confine ourselves to standard methods

and localities if resources and opportunity should ma'ke other methods

possible or should admit us to other localities. In fact, the excellence of

the general method has been shown more strongly in some supplementary

series taken by us than it has been in the standard series.

One series was taken in 1920 from Jacksonville, Florida to San Diego.

This was broken by rough weather in the Caribean Sea, but Crandall and

Michael, our most experienced plankton men, did the work and they both

emphatically stated that on that trip no series at all could have been

taken by any other method. Not long afterward a short series followed

by occasional catches over a period of six months was taken between

San Diego and Seattle from the Pacific Steamship Company's passenger

steamer Queen. None of these catches could have been obtained by any

other method. In both of these cases only three gallons of water was

filtered but even that small amount gave good results.

For the last three summers we have chartered a tuna fishing boat for

a few weeks work at two stations respectively five and ten miles seaward

from our pier. By use of the Kofoid closing bucket which holds five

gallons of water we have been able at these two stations to get for a time

daily catches at the surface and at twenty and forty meter levels. These

series differed from standard series only in the fact that samples were

taken from lower levels as well as from the standard surface level. All

alike were filtered through the standard net and handled in the standard

way. The marine experts of our staff are agreed that by no other method

than the dipping method could such satisfactory catches be so continuously

made or with so little equipment or at such small expense. For larger

plankton and for chemical examination other catches were made at these

stations with ordinary tow nets and closing nets.

In 1921 the California Academy of Sciences conducted some investi-

gations in the Gulf of California in connection with which a series of

diatom collections was made by the dipping method. A somewhat similar

series was obtained in 1922 in Pacific waters off Lower California through

182 W. E. ALLEN

the courtesy of the Mexican Government which was conducting a biological

survey of Pacific Islands. In the latter part of January 1923 the U. S.

Coast and Geodetic Survey steamer Pioneer began taking collections for

us which were continued from San Diego to Alaska. In none of these

cases would series of collections have been possible by customary methods.

Yet they are giving very excellent results.

Beginning with July 1922 a series of collections of zooplankton was

begun by the dipping method with filtration through the same net as was

used for diatoms. On account of smaller numbers of animals it was

necessary to filter one hundred liters of water instead of twenty-five.

This material was examined alive for identification and the contents

counted or estimated after killing by Dr. Helen E. Murphy. So far as can

be judged at present the results are just as promising as those for diatom

material. This is especially true concerning the larval forms which have

such great numerical prominence in the zooplankton.

The taxonomic and statistical work on marine copepods which Dr.

Esterly has been doing for several years has reached a temporary climax

in his recent publication of studies indicating seasonal distribution of

certain representative groups and in present efforts to segregate several

small species which have numerical importance but which are very difficult

to recognize under the low magnification necessary to counting.

The migratory habits of the copepods and some other plankton animals

indicate that they are more favorable subjects for statistical studies of

short period correlations than are the diatoms, and it seems probable that

the work done by Dr. Esterly constitutes a valuable foundation for sta-

tistical studies outlined by Dr. McEwen. Under such conditions the

prospect for advancement in copepod studies is limited only by the re-

sources and facilities which can be applied to such work.

In the last four years Dr. Essenberg has done some interesting work

on taxonomy and distribution of the Appendicularia, but present indica-

tions are that that group is not sufficiently prominent in our plankton

population to warrant continued special attention with our present limits

of facilities.

The results of our various methods of work in these somewhat different

but closely related lines have been very satisfactory. The most conspic-

uous features of these results are as follows:

1. All microplankton organisms show marked seasonal variations

in numbers. In higher levels of the sea Appendicularia are rare in the

warmer and sometimes very abundant in the colder months. Copepods

show larger numbers of adults in Spring and mid Summer with marked

abundance of particular species at other times. Diatoms are usually few

in mid and late Summer, most abundant in Spring, next most abundant

in Fall and sometimes prominent in late Winter. Dinoflagellates tend to

RECENT WOKK ON MARINE MICRO-PLANKTON 183

be most numerous in Spring, in late Summer and in Fall with some cases

of remarkable production in late summer.

2. Many zooplankton organisms, especially copepods, are strongly

migratory and usually negatively heliotropic. Hence, larger catches are

to be expected at or near the surface at night rather than in the day time.

There is evidence that tropisms may be reversed in response to changes

such as that of salinity but definite field work along that line is still lacking.

3. There is rapidly increasing evidence that the microplankton popula-

tion is unevenly distributed in most areas, both as to total numbers and

as to proportional representation of various species.

4. Present indications are that prominent plankton diatoms of one

oceanic area are likely to be found prominent in any area between the Gulf

of California and Puget Sound. Furthermore, it appears that most of

the important forms observed in Pacific waters are likely to be found at

some time of year in any part of this general region.

5. We have some evidence that heavy production of pelagic diatoms

is (south of Point Conception) characteristic of the sea rather than of bays

and lagoons.

6. Although heavy production of diatoms has been found nearly one

hundred miles from shore and near the limits of the continental shelf there

is good ground for thinking that the most productive areas, especially

below Pt. Conception, are within fifteen or twenty miles of shore, possibly

less.

7. We have considerable evidence that the most productive level for

diatoms is between ten and twenty meters below the surface.

8. Although surface levels are somewhat less productive than slightly

lower levels there is reason for thinking that general responses to environ-

ment are similar and that the trend of seasonal distribution is the same.

9. Conditions which cause heavy production of one or two species of

diatoms or dinoflagellates usually cause slightly increased production of

other species although these may be less conspicuous by comparison.

Furthermore, such favorable conditions also cause an increase in the total

number of species listed.

10. Although it is assumed that changes in temperature must have a

marked influence on all plankton organisms it has been found that heavy

production of diatoms may be very conspicuous at the same time at all

levels from the surface to thirty meters below in ranges of temperature

similar to those from December to June at the surface.

11. Our practical experience and our experiments have led to the

adoption of methods and plans for work on phytoplankton which seem to

be as satisfactory as the nature of the work will permit.

A KEY TO THE GENERA OF ACANTHOCEPHALA*

By

H. J. Van Cleave

University of Illinois

More than fifty generic names have been proposed for members of the

relatively small group of parasitic worms known as the Acanthocephala or

thorny-headed worms. Not all of these names are valid but there is no com-

pilation or analysis of these names which is of service to the general worker

in parasitology. Stiles and Hassall have made most valuable contributions

(1920) in their Index-Catalogue to the Roundworms but this work, which

is excellent for references prior to 1915, stops short of the period in which

the most active work has been in progress. Seventeen of the generic

names in the following list are more recent than the Stiles and Hassall

catalogue.

There have been continuous additions to the knowledge of the members

of this group since the time of the pioneer workers in parasitology dating

from the latter part of the eighteenth century but the systematic organiza-

tion of this information had its beginning only about three decades ago.

Prior to 1892, all species of Acanthocephala were, by most workers, placed

within the single genus Echinorhynchus. Beginning with Hamann's

contributions of that year, the number of generic names has increased at

a very rapid pace. No attempt has ever been made to prepare a key which

would make it possible for anyone other than a specialist, to recognize

the genera of specimens encountered in general collecting. A number of

workers have expressed the desirablity of having such a key and in the

belief that it would be of value the present one has been formulated. The

involved synonymy and frequent confusion of generic names has made it

seem desirable to preface the key by a tabulation of some of the more

important data concerning each genus.

In differentiating the genera of Acanthocephala, it is not always possible

to utilize external characters which are observable in living or preserved

specimens. Usually the specimens must be prepared for microscopic

examination. In following the appended key, well prepared toto-mounts

or sections are usually essential. In the preparation of permanent mounts,

many catastrophes and disappointments are avoided by puncturing the

body wall of each specimen with a dissecting needle while the specimens

are still in the preservative. Stains and the mounting medium enter the

body cavity and the internal organs much more readily in punctured

*Contributions from the Zoological Laboratory of the University of Illinois, No. 229.

1S4

A KEY TO THE GENERA OF ACAXTHOCEPHALA

185

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A KEY TO THE GENERA OF ACANTHOCEPHALA

187

specimens than in those with intact cuticula. Balsam or damar mounts

of specimens stained in dilute hematoxylin, especially Ehrlich's acid hema-

toxylin, are best suited for study. In some of the larger specimens, which

are too large for examination with the compound microscope, dissections

are essential. Synthetic oil of wintergreen as a clearing agent renders

even fairly large specimens remarkably translucent. Whole mounts

cleared in this oil and mounted in damar usually show all characters needed

for the identification of genera and often are sufficient for the determination

of species.

The accompanying figure (Fig. 1) shows the general organization of

the body of an acanthocephalan, though the individual organs are subject

to much deviation in form in the various genera.

Proboscis

Fig. 1. A male acanthocephalan of the genus Acanthocephalus.

The reference list at the close of this paper includes a number of the

more general works which are found useful in the determination of species.

1 (60) Anterior region of body bearing a proboscis upon which hooks or

spines are usually arranged in parallel alternating rows 2

2 (3) Region between body proper and proboscis (the neck) elongate,

cylindrical, except in region adjacent to proboscis where (except

in some young individuals) a conspicuous spherical enlargement

occurs. Genus Pomphorhynchus.

188 H. J. VAN CLEAVE

3 (2) Region behind proboscis variously modified but never bearing a

spherical enlargement followed by a narrow, elongated neck. 4

4 (41) Body proper never hearing spines. (Demonstration of minute

spines on the anterior part of the body in cleared specimens of

some genera requires careful adjustment of the light) 5

5 (14) Subcuticula bearing a few rounded giant nuclei, prominent in

stained whole mounts and in sections and their location frequently

recognizable in preserved specimens as minute elevations of the

body surface. Wall of proboscis receptacle with but a single

muscular layer. Cement gland of males a single syncitial mass

with a few giant nuclei. Family Neoechinorhynchidae. ... 6

6 (11) Proboscis globular, bearing three circles of hooks 7

7 (8) Six hooks in each circle of hooks upon proboscis. Genus Neoe-

chinorhynchus.

8 (7) More than six hooks in each circle 9

9 (10) Eight hooks in each circle. Genus Octospinifer.

10 (9) Twelve hooks in each circle. Genus Gracilisentis.

11 (6) Proboscis bearing more than three circles of hooks 12

12 (13) Twenty or more circles of hooks. Subcuticular nuclei all in

sagittal plane. Genus Tanaorhamphus.

13 (12) Eight circles of hooks upon proboscis. Giant nuclei of subcuticula

not all in sagittal plane. Genus Pandosentis.

14 (5) Subcuticula bearing coarsely branched or finely dendritic nuclei

or numerous small nuclei scattered through the subcuticula, but

never with rounded giant nuclei 15

15 (24) Less than eight cement glands in the male. Proboscis receptacle

sac-like, with two muscular layers 16

16 (19) Parasitic in birds. Males with three very long tubular cement

glands 17

17 (18) Proboscis receptacle inserted near middle of the proboscis wall;

the portion of the proboscis posterior to the insertion bearing

simple, thorn-like spines; anterior to the insertion bearing strong

hooks with proteriorly recurved, simple roots. Genus Cen-

trorhynchus.

18 (17) Proboscis receptacle inserted at base of a very long cylindrical

or clavate proboscis. Genus Prosthorhynchus.

19 (16) Males with six cement glands, though of highly variable form . . 20

20 (21) Parasitic in birds. Genus Plagiorhynchus.

21 (20) Parasitic in fishes, amphibians, or reptiles 22

22 (23) Brain somewhat in front of the posterior extremity of the pro-

boscis receptacle. Retinacula pass through lateral walls of

receptacle. Hooks with simple posteriorly recurved roots. Genus

Echinorhynchus.

23 (22) Brain at posterior extremity of proboscis receptacle. Retinacula

pass through posterior extremity of receptacle. In many species

lateral or anteriorly directed processes are given oil from the

main posteriorly directed root. Genus Acantliocephalus.

A KEY TO THE GENERA OF ACANTHOCEPHALA 189

24 (15) With eight cement glands. Parasitic in birds and mammals. 25

25 (26) Proboscis receptacle a closed muscular sac with retractors passing

through posterior extremity. Outer layer of receptacle disposed

in spiral bands. Hooks small, simple, each with a single simple

posteriorly directed root. Parasitic in mammals. Body showing

evidences of pseudo-segmentation. Genus Moniliformis.

26 (25) Proboscis receptacle a single-walled sac from a cleft in the ventral

surface of which the proboscis retractors continue posteriorly

through the body cavity. Brain inside the receptacle in region

of cleft 27

27 (30) Parasitic in birds 28

28 (29) Proboscis globular, armed with relatively small number of heavy

hooks, roots of some of which bear one or more branches directed

anteriorly. Genus Oligacanthorhynchus.

29 (28) Anterior region of proboscis provided with strong hooks, basal

region (sometimes erroneously termed the neck) with simple

spines. Each hook on anterior part of proboscis joins a posteriorly

directed root which at the union with the hook is elongated and

narrow but expands posteriorly into a much wider flattened,

circular, termination. Spines on posterior region of proboscis

not always in perfect rows but usually in twice as many long-

itudinal rows as the hooks on anterior region of the proboscis.

Genus Mediorhynchus.

30 (27) Parasitic in mammals 31

31 (34) Reproductive organs of the male confined to the posterior region

of body cavity. Lemnisci filiform, very long 32

32 (33) Proboscis provided with a crown of a few circles of strong hooks,

crowded so that they may have the appearance of a single row,

and behind this crown a region of some length closely set with

fine spines. Genus Gigantorhynchus.

33 (32) Proboscis with several circles of strong hooks. In intestine of

marsupials and edentates. Genus Hamanniella.

34 (31) Reproductive organs of male occupy more than one-half of the

length of the body cavity. Lemnisci frequently in contact with

testes 35

35 (38) Cement glands of male arranged in a linear series of four pairs. 36

36 (37) Lemnisci relatively short, flat. Testes considerably distant from

cement glands. Genus Macracanthorhynchus.

37 (36) Lemnisci relatively long, subcylindrical. Posterior testis not far

removed from cement glands. Genus Echinopardalis.

38 (35) The eight cement glands not definitely arranged in pairs 39

39 (40) Cement glands almost spherical. Lemnisci very long, sub-

cylindrical, more than three-fourths the length of the body

cavity. Genus Oncicola.

40 (39) Cement glands closely crowded together. Genus Prosthenorchis.

41 (4) Body-proper provided with at least a few spines (in some indi-

viduals difficult to detect even in cleared specimens) 42

190 H. J. ^"A^• CLEAVE

42 (45) Spines not limited to anterior region of body, some spines occurring

near genital orfice though in some gravid females the genital

spines may be lost with the shedding of the copulatory cap. . 43

43 (44) Body just behind the proboscis swollen, closely set with spines

and serving for an accessory organ of fixation. Proboscis of

medium length. Parasitic as adults in mammals and birds.

Genus Corynosoma.

44 (43) Anterior region of body slender, spines scattered. Proboscis

long. Parasitic in fishes. Genus Telosentis.

45 (42) Spines wanting on posterior extremity of body 46

46 (47) Body spines limited to a collar composed of four rows encircling

the body in the region of the proboscis receptacle. Wall of recep-

tacle but a single muscular layer. Parasitic in fishes. Genus

QuadrJgyrus.

47 (46) Body spines not limited to a few complete circles 48

48 (49) Body spines arranged as a collar near the anterior extremity

and behind this arranged in 18-23 cross-rows of closely set spines,

the rows separated from one another by considerable area devoid

of spines. Genus Serrasentis.

49 (48) Body spines not arranged in ventral transverse rows widely

separated from one another 50

50 (51) Parasitic in marine mammals. Spines thickly set as a continuous

mantle covering an enlarged anterior extremity of the body-

proper. Genus Bolbosoma.

51 (50) Parasitic in birds and fishes 52

52 (57) Hooks on ventral surface of proboscis distinctly larger and

heavier than hooks at corresponding level on dorsal surface . . 53

53 (54) Parasitic in birds. Proboscis usually enlarged near center,

spindle-shaped. Anterior region of body usually somewhat

enlarged, with its wall of different thickness and different histo-

logical composition than the wall of posterior region of body.

Genus Arhythmorhynchus.

54 (53) Parasitic in fishes. Body cylindrical or tapering gradually, at

least without an inflation near anterior extremity, sometimes

slightly swollen in region of body spines on ventral surface.

Length of proboscis many times the diameter. Body spines

ensheathed in prominent cuticular folds 55

55 (56) Body spines scattered, especially conspicuous on ventral surface

of body. Proboscis hooks protrude at least half their length

from surface of pr()l)()scis. Brain near middle of proboscis re-

ceptacle. Genus Rhadinorhynchus.

56 (55) Body spines form an uninterrupted mantle extending backward

from anterior extremity of body-proper. Brain near anterior

extremity of receptacle. Proboscis hooks protrude but a short

distance beyond the heavy ' cuticula investing the proboscis.

Genus Tegorhynchus.

A KEY TO THE GENERA OF ACANTHOCEPHALA 191

57 (52) Proboscis hooks of dorsal and ventral surfaces essentially similar

in size. Parasitic in birds. Anterior extremity of body set with

fine spines. Neck unarmed 58

58 (59) Proboscis ovoid or spherical. Body usually very thick. Anterior

extremity of body-proper covered with scattered spines. Between

this spined region and the proboscis a long cylindrical neck,

frequently retracted within the anterior extremity of the body.

In adult females of some species the proboscis is inflated as a

large sphere which bears a star-shaped arrangement of radiating

rows of hooks at the distal pole. Genus Filicollis.

59 (58) Proboscis relatively long, cylindrical or club-shaped, frequently

swollen at base; length about twice the diameter. Anterior spined

region usually set off from rest of body by a constriction of the

body wall. Genus Polymorphus.

60 (1) Hook-covered proboscis lacking. Anterior extremity of body

enlarged and bearing small pit-like depressions. Beneath the skin

around the anus of birds. Genus Apororhynchus.

References

LtJHE, M. 1904-5. Geschichte und Ergebnisse der Echinorhynchen-Forschung bis auf

Westrumb (1821). Zool. Annal., 1:139-353.

1911. Acanthocephalen. Die Siisswasserfauna Deutschlands, Heft 16. Jena.

Marval, L de, 1905. Monographic des Acanthocephales d'oiseaux. Rev. Suisse de Zool.,

13:195-386.

Porta, A. 1905. Gli Echinorinchi dei Pesci. Arch. Zoologico, 2:149-214.

1909. Gli Acantocefali degU Anfibii e dei Rettili. Ibid, 3:225-259.

1909. Gli Acantocefali dei Mammiferi. Ibid, 4:239-285.

Travassos, L 1917. Contribuicoes para conhccimento da fauna helmintolojica brazileiros.

VI, Revissao dos acantocefalos brazileiros. Parte I. Fam. Gigantorhjoichidae,

Hamann, 1892. Mem. Inst. Oswaldo Cruz, 9:5-62.

1920. Acanthocephalos dos animaes domesticos. Revista de Veterinaria e Zootechnia,

10:3-23.

Van Cleave, K. J. 1918. The Acanthocephala of North American Birds. Trans. Amer.

Micros. Soc, 37:19-48.

1919. Acanthocephala from the Illinois River, with descriptions of species and a synopsis

of the family Neoechinorhynchidae. Bull. 111. Nat. Hist. Survey, 13 (8):i-iv,

225-257.

1920. Two new genera and species of Acanthocephalous worms from Venezuelan fishes.

Proc. U. S. Nat. Musum, 58:455-466.

1921. Acanthocephala collected by the Swedish Expedition to the Juan Fernandez

Islands (1916-17). Natural History of the Juau Fernandez and Easter Island,

Part 13:75-80.

DEPARTMENT OF METHODS, REVIEWS, ABSTRACTS AND

BRIEFER ARTICLES

THE POCKET MICROSCOPE "TAMI"

It is a remarkable sign of progress and a proof of the never resting

spirit of the microscope manufacturers, that lately an important innova-

tion has been developed in the construction of a small microscope, the

purpose of which is to replace effectively the larger instruments with low

or medium magnification.

The idea of constructing such a small, compact and strongly built

microscope of excellent optics, at a price which is much lower than those

now in use, originated at the factory of M. Hensoldt & Sons, "Wetzlar,

Germany, who for the last 71 years have been producing high grade optical

instruments of military character and for scientific research.

Especially will the botanist, zoologist, entomologist, mineralogist,

biologist welcome this new model microscope " T A M I" which measures

4" in height and 1^" in width. The whole instrument is covered up

with a solid metal hood; total weight is 15 ounces. Its slender and smooth

shape is inviting to carry it in the pocket for outdoor's use.

"T A M I" magnifies 50x and any degree up to 225x by simply extending

the tube-length, without changes of objectives nor eyepieces. By unscrew-

192

DEPARTMENT OF METHODS 193

ing the lower objective system, the magnification ranges from 25x to

112Hx.

Illumination is furnished, for transparent objects, by a concave mirror

from underneath. The mirror and entire stage is quickly removable and

This illustration shows detached base. The polished glass stage protects the mirror

from dust and moisture.

the "TAMI" proper can be placed on extra large, opaque objects, giving

the observer a chance to move it ail over the surfaces of largest specimens

of rock, metal plates, wood, paper, etc., etc.

The small dimensions of "TAMI," its extra light weight, slender shape,

solid and compact construction, excellent optics, altogether are making

a serious scientific instrument available at a very considerable cost price

and will make it a desirable possession for a large circle of people.

A NEW POCKET MICROSCOPE

A great many American manufacturers produce miniature models which

are duplicates of their standard products and which can be used by young

folks. One of our largest optical plants has seemingly done a similar thing

in producing an extremely small microscope which when folded can be

placed in a leather case pocket size. But this microscope, miniature though

it is in size, has adjustments and magnifications equal to many standard

models.

194 DEPARTMENT OF METHODS

The construction allows telescoping of the draw tube and the use of

one or both elements of the divisible objective in such manner as to give

a wide range of magnifications up to 250x. With these magnifications it

can be used in examining a great variety of objects, transparent or opaque,

in the laboratory or especially in field work in botany, entomology, mineral-

ogy and general nature study. The magnification is sufficient for chnical

examinations, including blood counting, and due to its portability, the

instrument may be used at the bedside of the patient.

As mentioned before, in adjustments and operation the pocket micro-

scope resembles the standard models. It is fitted with coarse and fine

adjustments which work in easy fashion. The stage is provided with two

spring clips which hold the specimen; and the mirror, adjusting in two

planes, serves in its regular position under the stage to illuminate trans-

parent specimens. When detached from the mirror bar, it can be placed

on a pin at the side of the arm to illuminate opaque specimens.

The instrument is supported by a tripod, the three legs of which fold

together and swing back parallel with the tubes and ready to place in the

leather covered pocket case, which measures 5 x 23^ x 2^^ inches.

The microscope weighs 13 oz. and is finished in smooth, durable black.

The eyepiece and divisible objective are of high quality and the instru-

ment in every particular is made to the Bausch & Lomb standard. Already

the demand has been surprisingly great and with the popularizing of a

real microscope, the company predicts a sale beyond estimates.

A NEW DRAWING APPARATUS

As soon as there is a demand for certain research apparatus, it is found

that some manufacturer has predicted the need or perhaps has been work-

ing along with the user of such equipment and therefore knows the wants

of others in research. At any rate, it is certain that in these days apparatus

DEPARTMENT OF METHODS 195

is not found wanting for any modern use. So this is the case with a new

drawing apparatus recently developed which adapts itself to all types of

drawing with the special added feature of use in either vertical or horizontal

position.

The interchange from horizontal to vertical position is simple and the

advantages are great. The optical base, with feet having a spread of

10^ X 15^ in., supports the optical bed, carrying the illuminating unit

and the microscope, in either position, according to the preference of the

user.

When used in a vertical position therefore, the bed is raised perpen-

dicular to the base and is supported by a heavy bracket attached to the

base by means of four screws, which can be removed by an ordinary heavy

screw driver. The optical bed is fitted on the back (or bottom) with a

threaded stud, and two dowel pins on the base hold the bed in alignment

when the apparatus is used in a horizontal position. The bed is 25 inches

long and graduated so that the parts can be easily relocated when it is

desired to duplicate a certain magnification.

The illuminating unit is one of the simplified types with which the

6-volt Mazda lamp is used as the light source.

The condensing lens regularly used is a 60 mm diameter, double

convex lens in a spiral focusing mount, which is replaced by the aspheric

condenser when the equipment is used for micro projection or photomic-

rography, because of its superior spherical correction and corresponding

increased efficiency.

The microscope used has been especially designed for this equipment,

although a standard microscope may be substituted. It has rack and

pinion coarse adjustment and fine adjustment which works equally well

in either the vertical or horizontal position. The body tube is easily

removed to make possible the utlization of the large field, for which such

lenses as the Micro Tessars are corrected.

196 DEPARTMENT OF METHODS

The advantages of this new microscope are obvious. On both sides

of the stage holes are drilled for the stage clips, so that the slide, if desired,

can be placed on the upper side of the stage when the outfit is used in a

vertical position. By placing the slide on the upper or rear side of the

stage, the specimen proper is always brought into the same plane with

reference to the objective regardless of the variation in thickness of slide,

an important factor in reconstruction work where constant magnification

must be maintained. The arm carrying the substage condenser has been

of special length and makes possible the use of slides of large size, the dis-

tance from the supporting post to the center of the stage opening being

21^ in.

When the apparatus is used in a horizontal position for drawing, a

reflecting mirror is required to direct the beam downward at 90 to the

drawing paper. When the body tube and eyepieces are used, a small

mirror can be clamped to the eyepiece tube, but when the low power

objectives are being used, a much larger reflecting mirror is necessary to

intercept the very wide diverging beam of light. Such a mirror is also

suitable with an eyepiece, as the projection distance can be increased by

moving the microscope back toward the lamp with a corresponding

increase in magnification. The large mirror supplied is first surface type.

By removing the reflecting mirror this equipment can be used as a micro

projector with very satisfactory results at distances of 10 or 12 feet from

the screen. A three-sided metal shield is supplied, which protects the

drawing board to a considerable extent from extraneous light.

A very unique camera attachment can be secured for use in conjunc-

tion with this apparatus when photographs at moderate magnifications

are to be made. It consists of a sheet metal box attached to the base by a

bracket and two thumb screws, the bottom of the attachment resting upon

the table. The plate holder and focusing screen are at the bottom of this

box. The image is observed on the white focusing screen through the

opening at the top of the camera box; a light excluding metal slide closes

up this opening while the exposure is being made.

The many uses to which this apparatus can be put make it extremely

useful for general laboratory work.

LIST OF MEMBERS

Admitted Since the Last Published List

Arn, E. R 1 120 Fidelity Medical Building, Dayton, O.

Barrett, Harvey P 211 Vail Ave., Charlotte, N. C.

Beaver, Wm. C Wittenberg College, Springfield, O.

Covell, Walter P 2070 Monroe St., Corvallis, Ore.

Dice, Dora L 1109 E. University Ave., Ann Arbor, Mich.

Dorman, Henry 301 Nat. Hist. Bldg., Urbana, 111.

Elliott, Theodore S 5733 Kenwood Ave., Chicago, 111.

Fortner, H. C 89 N. Prospect St., Burlington, Vt.

Green, Bess, R 31 E. Daniel St., Champaign, 111.

Hagle, Leigh H Gregory, Mich.

Hayes, Doris Wanda 715 X. 32nd St., Lincoln, Nebr.

HoRTON, Ethel Sue South Dakota State College, Brookings, S. D.

JuDSON, Lyman 403 E. Cass St., Albion, Mich.

Miller, H. M Washington University, St. Louis, Mo.

Mossman, Harland W 1707 Jefferson St., Madison, Wis.

Pare, Mary Elizabeth 100 Oakland Ave., Marietta, Ohio.

Pearson, Norma T 208 N. Lake St., Madison, Wis.

Ritz, Wllliam a 2 Stone St., New York, N. Y.

ROUSH, Eva M. F 145 Beverly Ave., Morgantown, W. Va.

SCHULTZ, Leonard P 110 N. Berrien St., Albion, Mich.

Smith, Harvey McKinley Biology Building, Madison, Wis.

Wardlaw, Wm. D Pittsburgh College of Pharmacy,Pittsburgh, Pa.

Yancey P. Henry, S. J Colegio de S. Francisco Javie, Ona, (Burgos), Spain.

197

INDEX TO VOLUME XLII

Abnormal specimens of Helodrilus, 122.

Abstracts, 68, 129, 155, 192.

Acanthocephala, Key to Genera of, 184.

Allen, W. E., Recent work on Marine Micro-

plankton, 180.

American Microscopical Society, Proceedings

of, 76.

Apparatus, New Drawing, 194.

B

Beck, Wm. A., Illuminating Device for Micro"

scopes, 108.

Biological Station, La JoUa, 180.

Booth, Mary Allard, 73. _

Bowen, W. K., New Method for^Whole

Mounts, 156.

Briefer Articles, 68, 129, 155, 192.

Bryozoans, Entoproctan, Movements of, 135.

California, Occurrence of Evadne tergestina

in, 155.

Chandler, A. C, Life Cycle of Davainea prog-

lottina, 144.

Cladoceran, Marine, Occurrence of, 155.

Clench, Wm. J., The Use of Sodium Silicate

as a Mounting Medium, 69.

Creaser, Charles W., The Use of Sodium Sili-

cate as a Mounting Medium, 69.

Culturing Tubificidae, 155.

D

Davainea proglottina. Life Cycle of, 144.

Descriptions of twinned earthworms, 159.

Device, Illuminating, 108.

Diplocardia michaelseni, 175.

Dissecting Microscope, Pocket, 71.

Distribution of Frog Parasites, 79.

Douglas Lake Region, Frog Parasites of, 79.

Drawing Apparatus, New, 194.

E

Early Observations, and Primitive Micro-

scopes, 95.

Earthworms, twinned, 159.

Egg Laying Habits of Haminea virescens, 148.

Enchytraeidae, North American, 91.

Entoproctan Bryozoans, Movements of, 135.

Evadne tergestina, Occurrence in Southern

California, 155.

F

Fortner, Harry C, Distribution of the Frog

Parasites, 79.

Frog Parasites of Douglas Lake Region, 79.

Fungi, New Genera of, 43.

Galloway, T. W., Physiology of Reproduction,

72.

Genera of Acanthocephala, Key to, 184.

Green, Bess R., Abnormal Specimens of

Helodrilus, 122.

Guberlet, John E., Hemistomum confusum, a

Homonym, 68.

H

Habits, of Haminea virescens, 148.

Hague, Florence S., Studies on Sparganophilus

eiseni, 1.

Haminea virescens. Egg Laying Habits, 148.

Handbook for Beginners, Modern Microscopy,

133.

Helodrilus caliginosus trapezoides. Abnormal

specimens of, 122.

Helodrilus roseus. Abnormal specimens of, 122.

Hemistomum confusum, .\ Homonym, 68.

Hilton, W. A., Movements of Entoproctan

Bryozoans, 135.

Homonym, a, Hemistomum Confusum, 68.

Illuminating Device, 108.

K

Key to Genera of Acanthocephala, 184.

L

Laboratory, Methods of Culturing Tubifi-

cidae, 155.

La JoUa Biological Station, 180.

La Rue, Geo. R., Modern Microscopy, 133.

Life Cycle of Davainea proglottina, 144.

List of Members, 197.

Locy, Wm. A., Primitive Microscopes, 95.

M

Marine Cladoceran, Occurrence of, 155.

Marine Micro-plankton, 180.

199

200

INDEX TO VOLUME XLII

Members, List of, 197.

Method, New, for Whole Mounts, 156.

Methods, 68, 129, 155.

Methods of Culturing Tubificidae, 155.

Michaelseni, Diplocardia, 175.

Michigan, Frog Parasites of, 79.

Micro-plankton, Marine, 180.

Microscope, New' pocket Dissecting, 71.

Microscope, Pocket, 192.

Microscopes, Illuminating Device for, 108.

Microscopes, Primitive, 95.

Microscopy, Modern, Handbook, 133.

Modern Microscopy, 133.

Mounting Medium , Sodium Silicate as, 69.

Movements of Entoproctan Bryozoans, 135.

Murphy, Helen E., Occurrence of Marine

Cladocera, 155.

N

New Drawing Apparatus, 193.

New Genera of Fungi, 43.

New Method for Whole IMounts, 156.

New Pocket Dissecting Microscope, 71.

New Pocket Microscope, 193.

New Records of Enchytraeidae, 91.

North American Enchytraeidae, 91.

Note on Occurrence of Evadne tergestina, 155.

O

Observations on Davainea proglottina, 144.

Occurrence of Evadne tergestina, 155.

Parasites, Frog, 79.

Physiology of Reproduction, 72.

Pickett, F. L., Stability of Staining Solutions,

129.

Plunkett, O. A., Systematic Presentation, New

Genera Fungi, 43.

Pocket Dissecting Microscope, 71.

Pocket Microscope, 193.

Pocket Microscope "Tami," 192.

Powell, E. F., Methods of Culturing Tubi-

ficidae, 155.

Primitive Microscopes, 95.

Proceedings of the American Microscopical

Society, 76.

R

Recent Work on Marine Micro-plankton, 180.

Records of Enchytraeidae, 91.

Reproduction, Physiology of, 72.

Review, Modern Microscopy, 133.

Review, of The Physiology of Reproduction,

72.

Reviews, 68, 129, 155. 192.

Revision of description of Diplocardia mi-

chaelseni, 175.

Richards, A., Egg Laying Habits of Haminea

virescens, 148.

Ryan, Ruth W., Systematic Presentation of

New Genera of Fungi, 43.

S

Smith, Frank, Description of Diplocardia

michaelseni, 175.

Smith, Frank, Twinned earthworms, 159.

Sodium Silicate as Mounting Medium, 69.

Solutions, Staining, Stability of, 129.

Southern California, Occurrence of Evadne

tergestina in, 155.

Sparganophilus eiseni. Studies on, I.

Specimens, Abnormal, of Helodrilus, 122.

Specimens of Twinned earthwonns, 159.

Stability of Staining Solutions, 129.

Staining Solution, Stability of, 129.

Station, La Jolla Biological, 180.

Studies on Sparganophilus eiseni, 1.

Study of Entoproctan Bryozoans, 135.

Study of Stability of Staining Solutions, 129.

Systematic Presentation of New Genera oj

Fungi, 43.

T

Tami, Pocket Microscope, 192.

Titus, Bessie Perrault, Mary Allard Booth,

by, 73.

Tubificidae, Methods of Culturing, 155.

Twinned earthworms, 159.

U

United States, Davainea proglottina in, 144.

Use of Sodium Silicate as Mounting Medium,

69.

V

\'an Cleave, H. J., Key to Genera of .\can-

thocephala, 184.

W

Welch, Paul S., New Records of North

American Enchytraeidae, 91.

Whole Mounts, New Method, 156.

Young, P. .\., Systematic presentation of

New CJenera of lungi, 43.

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