Attention of members of the

AMERICAN MICROSCOPICAL SOCIETY

is called to the announcement of Grants

from the Spencer-Toller Research Fund.

(See TRANSACTIONS, July, 1913)

These awards are to stimulate and aid research in

Microscopic work

1. The Grantee must be a member of the Society

2. Resulting discoveries must first be offered for publi-

cation in the Society Transactions.

3. Application should be made to Dr. Henry B, Ward,

Chm., Urbana. Ill.

NOTICE TO MEMBERS

It is a source of regret to the Editor that the Transactions can-

not be issued as dated. The issue is dependent, however, upon the

‘acome of material from the members, and must await upon a certain

degree of balance in the material as well as quantity of material.

The Secretary will consider it a favor if members will notify

him of non-receipt of numbers of the Transactions, and of changes

of address.

T. W. GALLOWAY,

Secretary-Editor.

Beloit, Wisconsin.

TRANSACTIONS

OF THE

American Microscopical

Society

ORGANIZED 1878 INCORPORATED 1891

PUBLISHED QUARTERLY

BY THE SOCIETY

EDITED BY THE SECRETARY

VOLUME XXXVI

NuMBER ONE

Entered as Second-class Matter December 12, 1910, at the Post-office at Decatur,

Illinois, under act of March 3, 1879.

Decatur, ILL.

Review Printinc & STATIONERY Co.

1917

PP one

Beet a Th

evn

ape ee

apt LD

OFFICERS

BREDA LS Deed ABUSER. yb Vides bo ouie es COP UNG Oh ead Cieale wale Madison, Wis.

First Vice President: T.L. HANKINSON.........cccccecccccs Charleston, IIl.

second Vice Dresident:) Lo Be GRIPFIN ao sa eae viaic ead ons s Pittsburg, Pa.

MCT RLOe a he ONG CZAL COW AY iu DCN sw od etiam a aelelsta’s vis hcie edie b Beloit, Wis.

PPPOE Ae NAN CLDAVE 6 ov .cn od cine Co ed Melee vee REM CoS orate Urbana, IIl.

CrIOGtOH se) MAGNUS) PELAUM os\ui doles caus Weld hein dc aeee weg Meadville, Pa.

ELECTIVE MEMBERS OF THE EXECUTIVE COMMITTEE

RSOtIRGM ER AUR ATES 1 aly ul uie ie eee Geta ee ale ate a iete Mie ata lets Ann Arbor, Mich.

ieorse ESRI ale on Pot Gas OE Caras pharm rte Cult ca a EL ara Walla Walla, Wash.

EX-OFFICIO MEMBERS OF THE EXECUTIVE COMMITTEE

Past Presidents Still Retaining Membership in Society

R. H. Warp, M.D., F.R.M.S., of Troy, N. Y,,

at Indianapolis, Ind., 1878, and at Buffalo, N. Y., 1879

AvsBert McCatra, Ph.D., F. R. M. S., of Chicago, Ill.

Geo. E. Fett, M.D., F.R.M.S., of Buffalo, N. Y.,

at Chicago, Ill. 1883

at Detroit, Mich., 1890

Stmon Henry Gace, B.S., of Ithaca, N. Y.,

at Ithaca, N. Y., 1895 and 1906

A. Crirrorp Mercer, M.D., F.R.M.S., of Syracuse, N. Y.,

at Pittsburg, Pa., 1896

A. M. Buerte, M.D., of Columbus, Ohio,

at New York City, 1900

C. H. E1iceENMANN, Ph.D., of Bloomington, Ind.,

at Denver, Colo., 1901

E. A. Birce, LL.D., of Madison, Wis.

at Winona Lake, Ind., 1903

Henry B. Warp, A.M., Ph.D., of Urbana, IIl.,

at Sandusky, Ohio, 1905

HERBERT Osporn, M.S., of Columbus, Ohio,

at Minneapolis, Minn., 1910

A. E. Hertzyer, M.D., of Kansas City, Mo.,

| at Washington, D. C., 1911

F. D. Heap, Ph.D., of Philadelphia, Pa.,

at Cleveland, Ohio, 1912

CHARLES BrookKoVER, Pu. D., of Little Rock, Ark.,

at Philadelphia, Pa., 1914

Cuar_es A. Kororp, Ph.D., of Berkeley, Calif.,

at Columbus, Ohio, 1915

The Society does not hold itself responsible for the opinions expressed

by members in its published Transactions unless endorsed by special vote.

TABLE OF CONTENTS

FOR VOLUME XXXVI, Number 1, January, 1917

Bacteriology in Plant Pathology, with table of Species and Hosts, by

Fe Vs SHOVES oho sce y wee bests Lahin's bee dele bone bikes « SDR nn eal

Museum Methods, by C. C. Nutting.......... Me ee UAW Gus NN Renee aero

The Growth of Ameba on a Solid Medium for Class Use, by M. W.

RAV ELOEY Sho Se Shes bak gk ah AVALON CATER Ys pro sie CRA tw ha Ae a ose) Ae whee Bae

Notes and Reviews: An Apparatus for Gradual Dehydration, by G. H.

Bishop; An Individual Paraffin Bath, by Robert W. Henderson; A

Macroscopic Method of Reconstruction, by Robert W. Henderson;

An Easily Adjusted Embedding Box, by C. F. Dean; A Device to

Counteract the Effect of Vibration in Photo-Micrography, with

Plate I, by Alfred B. Hitchens; Photomicrographs of Crystallizable

Chemical Salts, with Plates II] to V, by Arthur W. Doubleday; A

New Mounting Medium, by C. E. Norton; A Method of Cutting

Planorbis Eggs, by A. Richards; Special Technical Methods Re-

ported from the Zoological Laboratory, University of Wisconsin, by

Thurlow C. Nelson; Evidence as to the Number of Segments in the

Head of Insects, with Plate V, by E. W. Roberts; Animal Micrology

(Guyer) ; Introduction to Neurology (Herrick)...............+4.-.

(This Number was issued on May 23, 1917.)

TRANSACTIONS

American Microscopical Society

(Published in Quarterly Installments)

Vol. XXXVI JANUARY, 1917 No. 1

DEPARTMENT OF SUMMARIES

TO BE DEVOTED TO DIGESTS OF PROGRESS

IN BIOLOGY

While the Transactions will continue to be primarily a Journal of research in micro-

biology, it is recognized that the field has become so broad as to preclude the possibility

of frequent articles in any one of the departments of special interest. Because of this

it will be the policy to present, from time to time, supplementary digests of the progress

being made in the various fields of micro-biology. It is also proposed to introduce similar

summaries of the progress made in some departments not represented in our articles of

research. This is done with the feeling that such reviews will increase the permanent

value of the Transactions to all who may not have access to a large list of technical

biological journals, nor the time to make the survey for themselves.

BACTERIOLOGY IN PLANT PATHOLOGY

By F. L. STEVENS

It is my purpose to direct your attention to the place and import-

ance of bacteriology in the field of plant pathology, from the stand-

point of North American students and investigators, and to

summarize briefly the progress during the comparatively brief in-

terval since the inception of this science. I shall first consider the

subject in its broader, general aspects, then in its more special

relations.

It was inevitable that the brilliant results of the early workers

in bacteriology, especially as regards fermentations and animal

pathology, should direct the attention of plant pathologists to bac-

teria as a possible source of solution of some of their own problems.

Thus, in the year 1877 we find Dr. Burrill saying regarding

pear blight. “The cambium of the blighted branch, when the trou-

ble first shows itself, and for some days thereafter, is filled with

minute moving particles.

ea F. L. STEVENS

“Not unfrequently, a thickish, brownish, sticky matter exudes

from affected limbs, sometimes so abundant as to run down the

surface or drop from the tree. This proves to be identical with

that noticed in the cambium, and unquestionably has the same origin.

The sticky, half-fluid substance thus exuding is entirely made up

of these minute oscillating particles.” Transactions Illinois Horti-

cultural Society, 1877, page 114.

In 1878 he said—“If we remove the bark of a newly-affected

limb, and place a little of the mucilaginous fluid from the brown tis-

sues under our microscope, the field is seen to be alive with moving

atoms known in a general way as bacteria. Sometimes a thick,

brownish fluid oozes from the bark of dying limbs and spreads over

the outside or falls in drops. This is apparently made up of living

things, myriads of them to be seen at once. A particle of this vis-

cous fluid introduced upon the point of a knife into the bark of a

healthy tree is in many cases followed by blight of the part, but

with me not in every instance.” Transactions Illinois Horticultural

Society, 1878, page 80.

In 1881 he said: “But of the pear trees inoculated by budding

and puncturing, as described, sixty-three percent became diseased,

exhibiting all the characteristics, externally and internally, of the

blight.

“Less than two percent of those not inoculated became infected

with the disease.

“The slight wounds made by the process of inoculation cannot

be charged with the results, for similar wounds were made with a

clean needle, and these rapidly healed without further injury. The

introduction of the virus introduced the cause of the disease, and

the potency of the virus was quite positively due to the living bac-

teria.” Blight of Pear and Apple Trees: 10th Report Board of

Trustees Ill. Ind. Univ., 69, 1881.

In American Naturalist, Vol. 15: p. 527, we find: ‘Certain

diseases of animals are now positively known to be due to the ac-

tion of the minute organisms commonly known as bacteria, but it

has not been shown that they also cause disease and death of plants,

except as recently announced by the writer in case of ‘blight’ in

pear and apple trees.

BACTERIOLOGY IN PLANT PATHOLOGY 7

“In 1877 I observed in the fluids of blighting pear trees, great

numbers of minute, moving things which were not clearly identified

as bacteria until the following year. Their presence was uniformly

detected in every examination made (and they were numerous)

during the summer of 1878, and the fact was reported to the IIli-

nois State Horticultural Society, in December of that year. Investi-

gations were not further prosecuted until June, 1880, when the

unusual prevalence of the disease called more special attention to

it. The same organisms, or those very similar, were as uniformly

found in the tissues of apple trees suffering with the disease called

twig blight. On diseased parts of both trees, drops of whitish, vis-

cid material were often found, oozing from the bark, and this proved

to be almost wholly made up of the bacteria.”

From the time of the early work of Dr. Burrill onward there

has been a steady increase in the number of diseases suspected or

-known to be of bacterial origin.

In. the early days, technique was undeveloped and many of the

early hints at bacterial diseases were indeed barely hints. Many

of the things described then are difficult or impossible of recognition

now, while others have become the subjects of repeated research

and knowledge of them has been clarified.

Here as elsewhere in science, fundamental battles were foun

In the period 1879 to 1892, many researches abroad and a few in

America were directed to determine whether bacteria occur in nor-

mal, sound plant tissues with of course a final result in the negative.

Though numerous plant diseases were early attributed to bac-

teria (1877-1883) the number which received final acceptance as

such grew slowly. The text books bearing upon the subject give

only grudging credence. Even as late as 1892 only 13 plant dis-

eases are given by Russell as of established bacterial origin with

nine “Probably of bacterial origin.” Marshall Ward, the English

authority, in 1894 editorially doubts even the bacterial origin of

pear blight. As late as 1897 Frank evinces doubt as to bacterial

diseases in these words: “Whether bacteria can be the cause of

disease in plants is always a question to be considered with circum-

spection.” p. 201 Frank Kampfbuch. But the most vigorous atti-

tude of denial was taken by Alfred Fischer in 1897 who held that |

ade

8 F. L. STEVENS

there are not and cannot be bacterial diseases of plants because

bacteria cannot enter plants except through wounds and that their

development then will be stopped by corky layers. “The uninjured

plant stands in open connection with the outer world only through

the stomata, which connection is so limited that the system of air-

filled intercellular spaces connects with the outer world but is

entirely closed to the cells. When bacterial germs are forced into

the stomata by wind or rain, they here reach only into these inter-

cellular spaces where nothing further is offered to them than vapor-

saturated air, where all nutrient substances are wanting, without

which no bacterial spore can germinate, no bacterial cell can mul-

tiply. All these peculiarities are wanting in the bacteria, against

which an uninjured plant is fully protected. But also the wounded

plant offers food for bacteria only in the opened, injured cells, a

source which is soon removed by the formation under the wounded

surface of an impenetrable cork layer (wound cork) which entirely

prevents any further flow of sap from the wound. The wound

does not remain moist, the injured cells shrivel and dry out, and

consequently the entrance of the bacteria is exactly so barred out

as in the uninjured plant. Consequently, there is not the least

danger of wound infections by bacteria, whose further progress

in the plant is also impossible.” Translation by E. F. Smith, Bac-

terial Diseases of Plants II, p. 15.

Wehmer in 1898 assumed a similar viewpoint. The trend of

ex cathedra opinion at this period toward the denial of the exist-

ence of bacterial plant diseases, in opposition to rapidly increasing

evidence of their abundance, appears to us today as most remark-

able. The controversy became crucial between Dr. Erwin F. Smith

and Dr. Fischer, the latter asserting in 1899 that “there has not yet

been published a single proof for bacterial plant disease which meets

all the requirements of exact bacteriology. Smith in 1899 and 1901

in masterly articles adduced such complete evidence that since then

his position has not been challenged. From that period to the pres-

ent time very numerous bacterial plant diseases have been described

in America and elsewhere until in 1915 we find more than 140

genera of host plants listed with something more than one hundred

BACTERIOLOGY IN PLANT PATHOLOGY 9

bacterial forms suspected or proved to be responsible for certain

of their diseases.

Owing to the difficulties and obscurities of the subject as com-

pared with that of fungous diseases, it is probable that a large per-

centage has been as yet overlooked, indeed the rapid growth in

number of descriptions of diseases of this class during the last

few years is especially striking and betokens corresponding in-

crease for some time to come.

In addition to mere listing and identification of the cause of

the disease, fundamental progress has been made in the knowledge

of the biological and ecological relations of the host and parasite.

The action of the parasite has by critical studies been shown

to be through enzymes and in particular enzymes acting upon and

dissolving the middle lamella and thus bringing about dissolution

of the tissues.

A group of diseases of distinct type, the “wilts” are found to

be largely, tho not exclusively bacterial, and are due to a plugging

or embolism brought about by growth of bacteria within the vessels

of the plant. Infection has been shown to occur in a variety of

ways, notably through wounds which break down the outer pro-

tective plant coverings or what is perhaps much more remarkable,

through natural openings such as stomata, water pores or nectaries.

Important contributions have been made upon the subject of

infection carriers, more particularly since the role of certain insects

in this regard has been ascertained with precision. Surface soil

water is in some cases responsible for extensive distribution of the

parasite. Continued growth and multiplication of parasitic bac-

teria in the fallen plant parts or even in the manure pile offers an

additional explanation of disease dispersal in some instances. In

other cases it has been demonstrated that the casual bacteria remain

alive upon the outside of the seed coats and thus lead to infection

of the ensuing crop.

Phytopathological bacterial studies have in several instances

been conducted with such thoroughness that descriptive and taxo-

_ nomic bacteriology have been distinctly enriched.

The bacteria involved in plant disease are preeminently of the

genera Pseudomonas and Bacillus. The Cocci, Bacteria and Spirilli

“ey

10 F. L. STEVENS

sO prominent in animal pathology sink to a very minor position,

there being no Spirilli known as plant pathogens and very few of the

genus Bacterium and still less of the genus Micrococcus.

The foregoing may serve to give a general impression of the

history and activities of students of plant pathology within the field

of bacteriology.

The following table shows most of the specific diseases that

have commanded attention in North America.

A straight, uncrossed line , indicates the date of first knowl-

edge of the bacterial nature of the disease, or in some instances

of the species of bacteria which cause it. One cross mark —|—,

indicates definite progress toward a cultural or morphological de-

scription, or both. Two cross marks —||—, indicate a rather

comprehensive bacterial study. Three cross marks —|||—, indicate

that quite complete bacterial study has been made. Two stars **,

indicate that the disease is one of much importance in this country ;

one star *, that it is important but not of first rank. Those with

no star are little known, or are of narrow geographic range or of lit-

tle import. Numbers in the table refer to the bibliography in Stevens,

“Fungi Which Cause Plant Disease,” beginning on page 53. “‘a.b.”

indicates that reference may be found in the “Additional Bibliog-

raphy” of the same work.

In general it is noted that very few bacterial diseases were

known prior to 1886 but that from then onward there has been a

steady increase in their number.

Studies in any way complete were very few prior to 1895 but

became much more abundant after that time and the newly de-

scribed diseases which take their places in our table in later years

frequently first appear before us with details well worked out. In

particular is this very satisfactory condition true within the last

decade and a half.

Time forbids discussion of these diseases, 64 in number on

more than 70 hosts. A few however demand special mention.

Bacillus amylovorus, the cause of the pome blight is widespread

and the cause of immense pecuniary loss.

Pseudomonas radicicola occupies the anomalous position of be-

ing a beneficial disease. The progress of knowledge concerning it

is not indicated in the table.

PARASITE

B. amylovorus

Ps. radicicola

B. ole

Ps. savastanoi

B. sorghi

B. zere

Ps. solanacearum

Ps. avens

B. avenze

B. catorovorus

Ps. campestris

Bact. michiganense

Ps. phaseoli

Micrococcus?

B. tracheiphilus

Ts. aroidexe

B. gossypini

Ps. mori

Micrococcus:?

Ps. stewartii

B. sp.

Ps. malvacearum

Ps. juglandis

Ps. amaranthi

BR. oleracea

Ps. woodsii

Ps. pruni

B. phytophthorus

B. delphini

Bact. teutlium

B. solanisaprus

Ps. tumefaciens

Ps. aptatum

Ps. medicaginis

B. araliavorus

B. muse

B. melonis

Ps. andropogoni

B. coli

Ps. maculicolum

Ps. cerasus

Ps. beticola

B. pyrocyanus

Ps. lachrymans

B. lathyri

Ps. citriputeale

Ps. erodii

Ps. veridilivium

Ps. citri

B. dianthi

B. fluorescens (?)

Bact. agropyri

TABLE SHOWING PROGRESS OF KNOWLEDGE CONCERNING AMERICAN PLANT DISEASES OF ECONOMIC IMPORT

HOSTS

Bas > eGR erates fits hits in Besser eps

SAO PAIMOOS Whe ininva flere pcos eld ova bials

AMSRRAS OLIVE Coos Ba cas cee

Olive

Salsify

PERFOR ER Ter aeiiin cxe inva Sisie.e ofa etn sym

Celery

Crucifers

Geranium

WOTEGOIS TIN oh. Sue. Metanemeleraicuniele s'als

Curcurbs

Carnation

Cotton

Mulberry

Gira w Derry wae save = slelel= <. o.0i0

CO CEOIN I Se eee teas santefernveieierers,, Is couslle

Juglans

Amaranthus

PSE WEG fale MiCneIPANT A © Sac aSeS ENCICL OR RCIEIACICIC On mORCnCn

Crucifers

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Prunus

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Delphinium

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Tropeolum, beet

Alfalfa

Ginseng

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Muskmelon

Broom COTM ....,ccccevcersces sons

Cocoanut, onion ........0+---

Cauliflower

Pepper

BeOGOMIG ic bicieis s opeisiere aiersie leaden

Cucumber ......- Sele einiel le/ejetere's

Curcurbs

Sweet pea, clover........ ae ar aie

Erodium, geranium .......... é

Lettuce ......++- Fis 3 Owen oc

1877 1878 1879 1880 1881 1882 1883 1884 1885

1899 1900 1901 1902 1903 1904 1905 1996

1886 1887 1888 1889 1890 1891 1892 1893 1894 1895 1896 1897 1898 1907 1908 1909 1910 1911 1912 1913 1914 1915 1916

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BACTERIOLOGY IN PLANT PATHOLOGY 11

Pseudomonas solanacearum is very wide-spread and destruc-

tive as are also Pseudomonas campestris and Bacillus tracheiphilus.

Bacillus carotovorus is the cause of soft rot of parenchyma on many

hosts and the loss of much produce during storage.

The bacterial leaf galls of the Rubiacez, originally described

by Zimmermann, are of particular interest as possible cases of sym-

biosis rather than parasitism.

Pseudomanos tumefaciens, the cause of crown gall on numerous

hosts, has been shown by Dr. Smith and his assistants in a series

of noteworthy investigations to furnish example of a here-to-fore

unsuspected type of plant disease, in many respects analogous to

human cancer. These investigations were prosecuted by overcom-

ing great technical difficulties of staining and culturing and yielded

results amazing from the viewpoint of anatomy and pathology and

suggestive and stimulative of investigation relative to human cancer.

Bacillus avene with Pseudomonas avene@ presents an unique

case of symbiosis in that the latter organism is, according to the

work of Manns, much more productive of disease when accom-

panied by the former organism.

Bacillus coli as the cause of a very destructive bud rot of the

cocoanut is especially interesting. One is loath to accept the con-

clusion but the work upon which it is based is well done. The ex-

perimental evidence is that the bud rot organism is in all ways

indistinguishable from animal Bacillus coli and that Bacillus colt

from animals will cause the typical rot. The case is still sub judice.

Pseudomonas citri is one of the latest to attract attention as

the cause of the very serious Citrus canker which bid fair to make

destructive inroads upon fruit culture in the Gulf states.

Several of the organisms or diseases listed above are so poorly

described as to be unrecognizable, thus it is probable that some of

these earlier ones are identical with those later listed as distinct and

definitely described species.

As to practical results in the way of prevention, the situation

is not so hopeful as in the case of diseases due to the Eumycetes.

The value of the knowledge is that knowing the cause and perhaps

the mode of hibernation and transmission, proper hygienic measures

may be practiced. Dispersal upon seeds may be avoided by seed

12 F. L. STEVENS

treatment; dispersal by insects by combating the insects directly.

Main sources of infection may be destroyed by fire, by the cutting

and burning of diseased parts; distribution through the manure

pile by care to avoid infecting the manure by diseased refuse.

In common with other fields of bacteriology there is need here

of fundamental knowledge of the biology of bacteria, their vari-

ability, methods of classification, a stable system of nomenclature, ©

of enyzmes, toxins, morbid histology, life histories, extra-parasitic

existence, persistence in soil, agents of transmission, climatic rela-

tions. The way is long, the difficulties numerous. Much has been

accomplished but much more remains to be done.

I cannot close this brief and manifestly inadequate presentation —

without paying especial tribute to the work of Dr. T. J. Burrill,

the deceased president of this society and that of Dr. Erwin F.

Smith, whose Treatise “Bacteria in Relation to Plant Disease”’

which though incomplete, already comprises three quarto volumes

that are models of completeness; whose numerous single papers

have enriched the world and whose careful judgment, and high

standards of work and presentation have done much to influence the

standards of all workers in this field.

(Read before the Society of American Bacteriologists at New

Haven, Dec., 1916.)

MUSEUM METHODS

By C. C. Nuttine

Curator, Museum of Natural History, State University of Iowa

It should be understood to begin with that we are dealing with

methods applicable to a zoological museum connected with a col-

legiate institution, a problem different in several important partic-

ulars from that of a municipal or state museum in that the primary

function of a college museum is to serve students, rather than the

general public. Systematic collections, for instance, are relatively

more important in a college museum than elsewhere; and enter-

tainment, in itself, it not of supreme importance although we can

by no means neglect it entirely.

Let us first consider some of the general considerations that

go far to make or mar any museum whatever, and none of these

is more vital than the problem of lighting.

When conditions permit, a combination of sky-lights and high

side-lights seems to me to be the best possible solution. In

“Mammal Hall’ and “Bird Hall” which ‘occupy the greater part

of the third story of our building,’ this plan has proved quite sat-

isfactory. Incidentally it is economical of wall space, as all of the

side walls can be occupied by exhibit cases up to a height of eight

feet. The light coming, as it does, from above this height is most

like that out of doors as to direction, and most of the reflections,

so distressing to the museum man as well as to the visitors, are

avoided. In some cases, where a uniform light is required and

special effects desired, the illumination should be entirely by electric

light which gives much more uniform effect than can possibly be

secured by the use of natural light. We have adopted this plan in

our “Laysan Exhibit” which is in the form of a cyclorama. An-

other valuable feature of this method is the fact that we can thus

1Hall of Natural Science, State University of Iowa. The writer has been requested

to discuss some of the methods used in the zoological museum of which he has been

curator for the past thirty years. Hence he feels free to refer to this museum as fur-

nishing illustrations of the points which he desires to make in this paper.

14 C. C. NUTTING

avoid almost entirely the bleaching effect on valuable specimens

that seems to be a natural concomitant of strong daylight.

If neither of the methods of lighting mentioned above is avail-

able, the next best thing seems to be the well known “alcove system”

so largely used in most museums.

The next general consideration, and one greatly neglected in

many museums, is the matter of a general color. scheme for a back-

ground for our exhibition series. We believe that this has been

solved in a very satisfactory way in our exhibits of mammals and

birds by a uniform background of dull light olive in all of the cases

for the systematic series, including the shelves. This color is very

restful to the eye, forms a sufficient, but not too glaring, contrast

with a vast majority of the specimens exhibited, and is, in fact, a

sort of impressionist effect most like that of out-of-doors, with its

green fields and forests, the commonest color in most landscapes.

It is the one the eye is most used to and therefore remarkably rest-

ful and satisfactory.

For invertebrate exhibits the problem is quite different. I sup-

pose no class of objects has been more nearly the despair of cura-

tors than the alcoholic collections, or those which have to be pre-

served in fluids. With us all such material is in cases built on the

alcove plan with a side light from large windows. The object here

is to make each individual specimen stand out clearly and without

distortion. We use square museum jars, of course. The general

background is a dull black, but each object has its own background

to suit its general color. These individual backgrounds gave us a

lot of trouble. The glass plates that are sold by dealers to put in

the jars are expensive and it is difficult to attach the specimens to

them. I have seen slate slabs used for this purpose, but these are

nearly as troublesome as glass. Finally we hit upon the idea of

plaster of paris plates which are made quickly and very cheaply by

pouring the thin plaster on a large sheet of plate glass and cutting

it inthe desired sizes before hardening. These cost almost noth-

ing, are practically indestructible, can easily be bored with holes

for the attachment of specimens and have a beautiful white sur-

face against which colored specimens stand out in a very satisfac-

tory way.

MUSEUM METHODS 15

But light colored specimens must have a darker background,

and we tried several methods of painting or staining these plaster

plates. Few paints or stains are unaffected by alcohol, formalin

and glycerin preservatives; and we were fairly stumped until we

tried Higgins’ indelible inks put on with a brush, which worked

beautifully. We can now have backgrounds to suit ourselves for

this very vexatious department of the museum.

Another important point in museum exhibits is to avoid over-

crowding. Anumber of objects, individually interesting and instruc-

tive, placed in close proximity is most confusing to the visitor and

forms an important contributory cause of “museum fatigue.”

We find that the best results are obtained by placing the square

jars some distance apart and mounting each specimen carefully

as a distinct entity, with nothing else sufficiently near to distract the

attention of the observer. Each jar has a label glued to its face

where it will not obstruct inspection of the specimen, and each label

is covered with a glass slip, a little larger than the label, securely

cemented to the jar. In other words, the label is practically a part

of the jar, can not be soiled nor easily displaced and is nearly in-

destructible, and each specimen is a separate focus of attention.

“Museum fatigue’ has been ably discussed by Benjamin Ives

Gilman in the ScrentiFIC MonTHLY, 2, p. 62. Having this in mind,

we avoid-table cases over which the visitor must bend, and use, so

far as possible, upright cases before which he stands erect. We also

attempt to so place specimens as to be within eighteen inches of

the level of the average eye. While this can not be entirely carried

out on account of enforced economy of space, it is possible to place

nearly all objects of real interest to the ordinary visitor in a hori-

zontal band of three feet, leaving less interesting or attractive ob-

jects to fill the space above and below this band.

Having considered the general questions of proper lighting,

backgrounds, spacing and avoidance of museum fatigue, we come

to a subject of grave concern to the curator in charge of the edu-

cational museum and that is the question of systematic series versus

habitat groups.

If the taxidermist, or “preparator,”’ as I believe he prefers to

be called, is an up-to-date man and a real artist he will throw all

16 CC) NUTTING

of his influence in the directioh of preparing the beautiful modern

habitat groups that are so justly admired by the public and so well

adapted to showing the artist’s ability and skill. And no one can

blame him for this, as it is the necessary expression of his creative

instinct. He has an actual horror of rows of birds on “T” perches,

arranged in solemn ranks and about as inspiring as the tin soldiers

of our infancy. Were the object of the museum simply to please

and entertain, his position would be unassailable. I contend, how-

ever, that the student of birds, for instance, should first of all be

able to identify at sight as many of the avian inhabitants of his re-

gion as possible. Habitat groups, beautiful and true to nature as

they sometimes (not usually) are, are necessarily too limited in

number and require too great an expenditure of time, money and

space to meet this primary educational need. On this account I

would have only so many habitat groups as can be prepared and

installed without sacrificing the systematic series, whose great edu-

cational function, it seems to me, justifies its retention in spite of

the opposition of the modern taxidermist, with whose point of view

we can still have a very real sympathy.

Some well prepared groups should, however, be in every zoo-—

logical museum. By this I do not mean the hap-hazard throwing

together of specimens and accessories that is such a painful feature

in too’many museums; but real pictures, correct and satisfying

from both the scientific and artistic points of view, such as are now

seen in most of our great museums and some of the smaller ones.

A compromise between the systematic series on “T” perches

and the habitat group idea has been attempted in some museums

and has attained a certain vogue. I refer to imitation or natural

branches, tree trunks, etc., with leaves and other accessories, each

with its bird or pair of birds with perhaps a nest, all fastened to

the common background of the case. Like most compromises this

arrangement is unsatisfactory, to me at least. It is “neither fish,

flesh nor good red herring.” The result is a sort of patch-work

resembling the crazy quilt of our youth. The specimens are usually

crowded together to economize space, and even if the separate pieces

are well done there is a painful sense of confusion as the eye tries

to take in too many details at once. On the other hand, the pri-

MUSEUM METHODS ig

mary object of the systematic series is lost because the specimens can

not easily be handled nor removed from their cases.

In the Hancock Museum on Newcastle-on-Tyne is the best ex-

hibit that I have seen along this line. It consists of walls com-

pletely occupied by small box cases of uniform or interchangeable

sizes, each with a pair of birds with well chosen accessories and a

separate painted background. I do not think, however, that even

this is as available for a college museum as the old-fashioned series

on uniform perches, with no atempt whatever at accessories, but in-

terspersed here and there with a few really good and well chosen

habitat groups.

The question of securing material for our exhibit might well

have been discussed earlier; but the general considerations already

touched upon should, in my opinion, be settled before the collections

are secured. How a museum should not be built up is sadly illus-

trated by a majority of American institutions. These are the re-

sult of gradual accretion of well-meant but disastrous donations of

specimens and collections from amateur collectors and “taxider-

mists” who have acquired a heterogeneous lot of material of which

they have become tired and whose room is better than its company.

Individually and collectively such aggregations are usually of

little value to an up-to-date museum. The taxidermy is ordinarily

atrocious and the data deficient or wanting. Often they are re-

garded with egregious pride by their donor or his heirs.

To decline such collections is apt to land the curator in high

disfavor with people who, after all, believe that they are doing a

highly laudable thing. Nowhere else is a superior diplomacy more

necessary than here, where the conscientious museum man is called

upon to avoid serious offense to friendly people on the one hand

and disaster to his exhibit on the other.

The best, and ordinarily the only, satisfactory way to secure

material is to go after it yourself, or send thoroughly competent men.

These men should not only be naturalists, but highly trained col-

lectors. As a general thing the man who is eventually to mount or

otherwise put the collection on exhibit should have as free a rein

as possible. If habitat groups are contemplated a competent artist

should go along to prepare sketches and details for the background.

18 Cc. C. NUTTING

If he and the taxidermist can properly “hit it off” together, the best

results will be attained.

Perhaps I may be permitted to refer once more to our Laysan

Island exhibit, the most ambitious and satisfactory thing that we

have attempted. After the expedition was organized, and financed

through donations from students and friends, it was placed abso-

lutely in the hands of Professor Homer R. Dill who finally prepared

the exhibit. We were also most fortunate in securing the services

of Mr. Charles A. Corwin, whose backgrounds in the Field Columbi-

an Museum have been so unusually beautiful and satisfying, as a

member of the party. An assistant taxidermist and general utility

man were added. The amount of work done and material secured

by this party of four men in the short time that they were permitted

to remain on the Island was astonishing. Accessory material, such

as bushes, samples of coral rock, leaves, grasses, entire nests of many

species, an ample series of sketches in oil by the artist, hundreds of

photographs and a collection of bird skins most beautifully prepared

in spite of the haste and pressure under which the work was accom-

plished ; all these were secured, packed and transported to their des-

tination without appreciable damage.

Then the masterly work of Professor Dill could be built upon

a proper foundation of ‘field observation, and the hearty and ef-

ficient co-operation of Mr. Corwin in painting his beautiful back-

ground representing the wind-swept mid-Pacific Island with its

surrounding reefs and Ocean has resulted in a piece of work that

is a delight from the standpoint of both naturalist and artist. This

background is 12’x138’, forming a cyclorama in which the back-

ground and foreground is joined with consummate skill through

the combined artistry of these two men.

The modern museum must have a taxidermist (or preparator,

if you will) not only up to date in the technic of the art, an artist

not only in skill but in feeling; but he should be a man able to ad-

vance the work by original ideas. In our museum that man is Pro-

fessor Dill. He has prepared, for instance, several mammal groups

that differ in conception from previous work. Such are the Moun-

tain Lion group in which the observer is inside the den, looking out

of the rugged entrance upon an alpine scene of quiet beauty illum-

MUSEUM METHODS 19

inated by concealed electric lights ; and a “‘flash-light” picture of Vir-

ginia Deer by a quiet woodland pool on a frosty night.

Apparently trivial details will often determine the success or

failure of a group. Snow and water must not look like frosting

on cake or glass or glue. An effective composition to fill out and

sustain the ears of large mammals must be light and hard, but not

brittle. One of the most difficult classes of objects to make pleasing

and natural is mounted fish and particularly their fins. The de-

greasing of old and more or less “burnt” skins without injury has

long been a problem. These difficulties, and others, have been suc-

cessfully surmounted in our case by Professor Dill who will, we

hope, some day give to the world the details of his methods.

Many points, such as the importance of securing and properly

storing the reserve or study series, the whole matter of satisfactory

labeling, the question of steel versus wooden cases, the proper ex-

hibition of small objects such as mollusks and of microscopic forms

such as Foraminifera, label holders, museum records, etc., must

be left untouched; for this article has already claimed too much

space.

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THE GROWTH OF AMCEBA ON A SOLID MEDIUM

FOR CLASS, USE

By M. W. WEtcH

In December, 1914 the class in laboratory methods at the Uni-

versity of Michigan was studying the methods of isolating and cul-

turing for class use, various living forms, especially protozoans.

The amceba proved to be the most difficult. It was not practical

to isolate in order to start a pure culture, and those cultures which

were made, were more or less uncertain, both as to the number of

amoeba and the time they would last. Dr. George R. La Rue, the

instructor of the class, suggested that it might be possible to grow

some forms for class use on a solid medium.

The preliminary work was just under way when Dr. James

B. Pollock, in isolating Azotobacter from the soil, found a large

number of small amcebe in his cultures. These were examined and

cultured on various media. Doctor Pollock pointed out that Azoto-

bacter would be a good organism to use for feeding amcebe, as

this bacterium can be cultured on nitrogen-free media. Such me-

dia would not readily become contaminated with many fungi and

bacteria which had been giving trouble in the previous work. This

has been the essential key to all future work. A summary of the

experiments on culturing the soil amcebe was read before the Michi-

gan Academy of Science, 1915 by Miss Kniseley, who had carried

on the work as a class problem.

In May, 1915 the work of further culturing amcebe was as-

signed to me, as Miss Kniseley had left the problem unfinished.

On May 20th inoculations were made onto ten slants of Ashby’s

Agar Medium (Ashby, 1907: 54) from the inactive tubes of Miss

Kniseley. In three to five days these all showed large numbers of

very active soil amcebe. In twelve to fourteen days practically all

of the amcebe had become encysted. All later observations have

borne out these results. Further work has not been done on the

life history of this ameeba.

22 M. W. WELCH

The object of my work was to find some form suitable for

class study which associated with an easily attainable bacterium

would grow on a synthetic medium. Many attempts were made to

isolate large amcebe from aqueous cultures and to inoculate the

soil amoeba cultures with them. None of these proved successful,

and in June preparations were made to keep the cultures running

over the summer. For this purpose a new medium was selected.

This was suggested in a paper by Miss Mockridge (1912:871). The

formula used was a modification of the formula given in her paper

for growing Azotobacter. The medium as used is as follows:

De@Xtrine i) hse cipteealele ap WM eittnINN Wlnce as elo 9 9 10.0 grm.

Di-potassium phosphate .......-.++seeereeee 2.0 grm.

Magnesium sulphate .......-+.-eeeeee rere 0.2 grm.

Calcium carbonate ......eee essere ener ee tees 0.2 grm.

Agat agar”. --a¢acreap ee eeinamemnec rs tsb os 10.0 grm.

Distilled water .....ceccececaccccercecseeeces 1000.0 c. c.

It should be noted that this formula is for a nitrogen-free nutri-

ent solution. This character makes it practically possible to inocu-

late from aqueous cultures without serious contamination.

During the summer the slants and cultures were taken to Lane

College, Chicago and attempts to isolate large amoeba and inocu-

late onto the cultures were continued. An aqueous culture obtained

while there, proved to be very plentifully filled with amoebze—five to

ten in most low power fields. Attempts to isolate and inoculate

from this culture proved unstccessful. A rougher method was

then tried. A few drops of the aqueous culture were put into each

of ten of the tube cultures of Azotobacter and soil amcebe, In

twenty-four hours two of the tubes were plentifully covered with

the larger amcebze, and in three days all showed excellent growths.

These ten tubes remained active for sixteen days to two months.

Transplants from these cultures in an active state gave new growths

of Azotobacter, soil amcoebz and large amoebz. Various other forms

that were in the original aqueous cultures have disappeared after

numerous transplants. Now, after 270 transplants and selections,

the cultures apparently show only bacteria, a tiny fungus, soil amce-

bee, and the larger amcebe.

THE GROWTH OF AMCBA yf

This large amoeba is very excellent for class use. When ex-

tended it measures from 90p to 150u. In general appearance it is

very much like Ameba proteus, but it is slightly smaller, and its

pseudopodia are frequently sharper than in the most typical form

of Ameba proteus. It is readily mistaken for Ameba proteus until

it is killed and stained, in which case the nucleus is found to be

round with a-clearly marked nuclear membrane. In the cultures

the amcebz are quite active, as well as being numerously distributed.

One small platinum loop from the culture may have from 50 to

200 or more amcebe. One tube will furnish sufficient material for

a class of 40. These cultures have been used with good success

this year at the University of Michigan and in various high schools.

Once started, it is very easy to keep cultures of this amceba

going indefinitely, thus assuring plenty of amcebz for class use at all

times. The cultures are kept as any ordinary bacterial culture, no

special methods being necessary. Transplants must be made while

the amcoebe are growing, and preferably while they are numerously

distributed over the culture. They may be kept at ordinary room

temperature, but must not be exposed to direct sunlight. It is best

to keep ten cultures going in order to insure a good quantity in

some of the tubes. A life history of one of the tubes will show

how often transplants have been made; these were usually made

from the tubes having the most amcebz present in them.

Date of original inoculation from aqueous cultures, 6/30/15.

Tube Nos. aR NA Source

II G. 5/10/16 II N

II N 4/5/16 IIA

IIA 3/8/16 II C

II C 2/15/16 IIA

IIA 12/17/15 Fr II;

Fr II; 11/30/15 Fr 11,

Fr II; 9/21/15 Fr II

Two plans of procedure for cultivation of amcebe are here

given.' The work should be carried on by one who is at least slightly

familiar with methods of culturing bacteria or fungi. The first

24 M. W. WELCH

is given as a guide to a rapid method for starting and maintaining

the cultures.

1. Prepare nutrient agar slants in test tubes, using the formula

given heretofore.

2. Obtain a culture of the ameebe.’

3. Make transplants onto ten tubes.

4. Keep at room temperature out of direct sunlight. Examine

after four or five days.

5. Examine in five to eight weeks and make ten new transplants

from the best tube. By continuing this method an abundance

of amcebe will be assured at all times.

The second plan of procedure is much more comprehensive

and is intended as a guide to those who care to start the method

from the beginning.

ISOLATION OF AZOTOBACTER FROM THE SOIL

1. Select five grams of poor soil and shake with about thirty

cubic centimeters of distilled water.

2. When the soil has settled, inoculate five drops of the liquid

‘nto each of ten tubes containing the solution heretofore given,

omitting the agar.

3. After three to ten days examine the tubes for Azotobacter.

From the tube showing the best growth, inoculate into three tubes

containing liquid agar of the given nutrient solution, at 42° C, in

the following manner: a. Three loops into the first of the three

tubes. Shake well. b. Three loops from the first tube into the sec-

ond and shake. c. Three loops from the second tube into the third.

(Reference to almost any text book on microbiology will give

the general methods for preparing agar slants, and for isolating

bacteria by the poured plate method. “Microbiology” by Giltner,

Pub. by Wiley and Son is recommended. )

4. Pour the contents of the three tubes into three sterile petrie

dishes. Keep the dishes in a cool dark place.

5, After the spot growths have started, examine them for

Azotobacter, and make inoculations onto agar slants from the spot

1The author will be glad to supply cultures to investigators interested in examining

the method. Others may buy them from Mr. W. J. Johnson, 1806 Morse Ave., Chi-

cago, Ill.

THE GROWTH OF AMCBA 25

showing pure Azotobacter. (If any show soil protozoa, it would

be well to make inoculations from them also, as the amcebe seem to

thrive better when associated with them. )

OBTAINING AMG@BZ

6. At the same time that the work of isolating Azotobacter is

being started, take steps to obtain ameebe. Jenning’s (1903: 2406)

method of culturing amcebe and other protozoa for class use is

recommended.

7. When the tubes containing Azotobacter are ready, and a

culture plentifully supplied with amcebe is present, inoculate each

of the ten tubes of Azotobacter with a few drops from the aqueous -

culture. This step may have to be repeated a number of times from

different cultures until an amceba which will feed on Azotobacter

is found.

8. The cultures may be maintained by following steps 1, 3, 4,

and 5 of the first method.

PAPERS CITED

Asupsy, S. F.

1907-8. Journal of Agricultural Science. 2:54.

JENNINGS, H. S.

1903. Methods of Cultivating Amceba and Other Protozoa for Class

Use, Journ. Appl. Microsc. and Lab. Methods, Rochester.

6 :2406.

MOocKRIDGE, FLORENCE A.

1912. Some Conditions Influencing the Fixation of Nitrogen by Azoto-

bacter and the Growth of the Organism. Annal. Bot. 26:871.

DEPARTMENT OF NOTES, REVIEWS, ETC.

It is the purpose, in this department, to present from time to time brief original

notes, both of methods of work and of results, by members of the Society. All

members are invited to submit such items. In the absence of these there will be given a

few brief abstracts of recent work of more general interest to students and teachers.

There will be no attempt to make these abstracts exhaustive. They will illustrate progress

without attempting to define it, and will thus give to the teacher current illustrations, and

to the isolated student suggestions of suitable fields of investigation.—[Editor.]

AN APPARATUS FOR GRADUAL DEHYDRATION

In working with delicate tissues, the more gradual the exchange

of fluids the less likely are osmotic distortions to occur. The ap-

paratus here described was devised to bring about an exchange of

fluids using the minimum amounts, without abrupt transitional

changes, while at the same time controlling the rate of change from

one density to another.

In Figure 1, an outer tube (a) carrying a small bulb below it,

receives an inner tube (b) fitting the outer rather closely for some

distance from the top, and tapering below to a capillary of about

Imm. diameter. This capillary (c) is recurved at the end to pre-

vent back-flow. The bulb (f) and the stem below it are packed

with cotton or other porous material to insure mixing of fluids

passing thru. A short length of rubber tubing (g) connects the

stem below the bulb with a glass tube (h) drawn out to a capillary

point, and thrust into the tubing point upward. The size of the

capillary opening of this tip determines the rate of flow. Its in-

sertion into the tubing point upward renders clogging improbable,

and eliminates any resistance that capillary action in this end-piece

might occasion. A container for tissues (i) with its overflow (k)

completes the apparatus. The whole is supported by two clamps

on a ringstand, one holding the tube (a) the other the container (i).

The overflow is caught in a waste receptacle below.

Let us suppose we wish to pass bits of tissue from 50 to 95%

alcohol. A series of pieces suspended on a thread or placed be-

tween loose plugs of cotton are held in a container (i) of minimum

length and diameter that will accommodate them. The smaller the

‘|

NOTES, REVIEWS, ETC.

DEHYDRATION APPARATUS

AMERICAN MICROSCOPICAL SOCIETY 29

diameter of the container the faster will the current of alcohol move

past the tissues, and the more quickly remove waste products soaked

from them. A plug on the end of a wire is inserted from the top

of the apparatus into the taper near the bottom of (b), and 50%

alcohol is poured in. Enough should be allowed to flow past the

plug, (by raising it slightly) to fill the bulb (f) up to the bottom

of the outer tube. Ninety-five per cent alcohol is then poured into

the outer tube at the lip on its upper edge, until full. The amount

of 50% placed in the inner tube should be a very little more than

enough to balance the column outside, since the two are to be in

equilibrium during operation. This amount may be ascertained

by trial or by computation, and indicated by a scratch on the glass.

It may be computed by figuring the height of a column of 50%

alcohol in the inner tube that will balance the outer tube filled full.

Let this height be represented by (y), that of the inner column by

(x). If X is the density of 50% alcohol and Y of 95%, then

xX=yY, and xy. If a little more than enough to balance is

placed in the inner tube it will do no harm, as it will flow thru the

capillary and remain beneath the lighter fluid. If the other flows

back however it rises thru the heavier liquid and contaminates it,

and further disturbs the equilibrium by rendering the column of

fluid thru the capillary too light.

The first fluid issuing from the apparatus will be pure 50%

alcohol from the mixing bulb. As the liquid is drawn off, and the

levels of the two containers lower, (the two columns remaining in

equilibrium,) the proportion of the heavier alcohol drawn from the

inner tube will decrease from 100% to 0, while the proportion of

lighter alcohol drawn from the outer tube will increase from 0 to

100%. The resultant liquid dropping into the container after being

mixed in the cotton packed bulb will consist of a perfectly graduated

column of alcohol varying from 50% to 95%, which will move

slowly past any tissues that may be suspended in the container.

After the apparatus has run dry, a little more 95% alcohol

dripped thru will force the graduated column, or the last end of it,

past the tissues, and leave them in pure 95%. Without removing

30 NOTES, REVIEWS, ETC.

from the container they may be put thru further changes with other

fluids.

To pass tissties from a higher to a lower alcohol, or from a

light to a dense fluid in general, requires a modification of this ap-

paratus. If the heavy fluid were placed in the outer tube, to reverse

the process as described above, the content of the inner tube, as it

proceeded from the capillary opening, would rise thru the heavier

liquid without mixing. One of two methods can be employed to

prevent this. Either the taper can be blown in the opposite direc-

tion, large and tight fitting at the bottom and small at the top; or

the method of delivering the two fluids into the mixing chamber

may be modified.

Figure 2 shows the apparatus with a cork (e) fitted into the

neck between tube and mixing bulb. A piece of rubber tubing (j)

‘5 fitted thru the center of the cork, and a recurved capillary tube

(d) thru one side of the center. For greater facility in cleaning,

replacement of parts, and construction or readjustment of the ap-

paratus the outer tube (a) and-the mixing bulb (f) can be made

separate and held together by inserting the cork into the end of each.

The ideal construction would by a ground joint such that the upper

part would fit into the lower as a ground glass stopper fits a bottle ;

the cork being first inserted into the bottom of the upper section.

The lighter liquid is placed in the inner tube, which fits tightly into

the rubber tubing thru the cork. Enough is allowed to run thru

‘nto the bulb to fill this as before. The outside tube is filled with the

heavy liquid, to a height to balance the inner column. The tubes

are made to fit tightly for some distance from the top, and the taper

of the inside tube should begin at the level to which, when the appa-

ratus is used to pass from lighter to denser fluids, it will be necessary

to fill the outer tube. (This will not hinder its operation in the

reverse process.) The tissue holder must be reversed so that the

flow is in thru the smaller arm and out at the top of (i). This arm

(k) should not be more than a millimeter or two in diameter.

The contents of the mixing chamber, being of the lighter fluid,

may come down after the upper part of the apparatus is exhausted,

and contaminate the tissues again. Either the apparatus must be

watched, or the arm (k) made so little higher than the container

AMERICAN MICROSCOPICAL SOCIETY 31

that when the arm fills with the lighter liquid it will overflow before

it forces any of the lighter liquid to the inlet at the bottom. When

the apparatus is empty a little more of the heavy fluid dripped thru

as in the former operation will replace the last of the column of

mixed fluids.

Several points should be noted in the construction of the ap-

paratus.

The inner tapering tube is not blown as a straight cone, but

flares at the middle. The curve of its side may be computed simply.

Let D be the diameter of the tube at the wide top. The volume of

a short section of the tube at that region is D? times a constant. If

volumes of such sections are to be made to decrease equably from

top to bottom, to obtain a change in the relative amounts of the two

liquids proportional to the time of operation, the diameters corre-

sponding to these decreasing volumes in the inner tube must de-

crease as the square root of the volumes. So the diameter half way

down must be the square root of = ; that 34 of the way down the

square root of oa that 14, the square root of 32, and so forth.

For practical purposes the volume of the glass itself may be dis-

regarded. ,

Unless the point of this tapering tube is recurved, to form a

sort of trap, (also that of the capillary tube (d) thrust thru the

cork at (e),) the lighter liquid will diffuse upward thru the capillary

containing the heavier as soon as the flow thru the latter becomes

slow.

If the tubes fit too tightly at the top, surface tension between

them may result in an interference with the balance between the

two fluids. This can be obviated by blowing the outer tube with

a channel running from the lip provided for filling down the side

to a distance beyond the beginning of the taper of the inner tube.

A number of different capillary tips to fit into the rubber tubing

at (g) may be provided, and the flow regulated by a choice of tips,

or a clogged tip replaced. While the tip is being removed the flow

can be stopped by pinching the tubing between the fingers. Other

devices for controlling flow, such as pinch cock, stop cocks, etc.,

were tried, but none is so satisfactory as the inverted capillary.

No mention has been made of size, because the varying needs

32 NOTES, REVIEWS, ETC.

of the user determine this. Several would be needed for any gen-

eral work. One now in use, holding 500cc., empties in five to six

hours, dropping alcohol, at the rate of 50 drops per minute. This

is very rapid, and the same apparatus can be made to run ten to

twelve hours at least. Another containing 40cc. runs two hours,

dropping at the rate of five drops per minute.

The tubes (a) and (b) must not be too long in proportion to

diameter and desired flow, since the taller the apparatus the greater

the difference in pressure between start and finish, which means a

difference in rate of flow correspondingly, (tho not proportionally)

great. A proportion of length to diameter of 3 to 1 has been found

most practical. A broader one is disturbed by jarring of the ap-

paratus, and a taller one varies considerable in rate. The mixing

chamber should be at least an inch long, better an inch and a half,

but may be of small diameter.

If the fluids should be imperfectly mixed when they issue from

the apparatus, the unmixed portions will almost immediately. ar-

range themselves in the container and no harm result. With a

properly packed mixing chamber 95% alcohol and water may be so

mixed that the resultant fluid shows no diffusion currents in the

container. It has usually been found desirable however to use two

fluids nearer together in density, as water and 50% alcohol, then

50% alcohol with 95% alcohol, then 95 and 100% alcohol, then

100% and a mixture of half 100% and half xylol or other oil, etc.

The result is greater certainty and better efficiency in the use of

the apparatus. |

G. H. BisHop.

Zoological Laboratories, U. of Wisconsin.

AN INDIVIDUAL PARAFFIN BATH

An improved form of the Self-Regulating Paraffin Bath as de-

signed by C. W. Woodruff, and described in the Journal of the Royal

Microscopical Society for October of 1914, has been in use in the

Animal Biology Laboratories at the State University of Iowa for

nearly three years.

The original apparatus was made of tin, but as tin rusted so

quickly other materials have been substituted. So, now the appara-

tus consists of a glass double cup blown in one piece, one cup be-

AMERICAN MICROSCOPICAL SOCIETY 33

ing within the other separated by a distance of approximately one

centimeter at the sides and two centimeters at the bottom. In addi-

tion to this cup is an ordinary 16 candle-power electric bulb, a con-

denser, an asbestos container, and Acetone which is the solution

used as the bath.

The inner glass cup is just large enough to contain an 100 c.c.

aluminum cup which is removable and in which are placed the par-

raffin and tissues to be imbedded. Good thermal contact between

the inner glass cup and the aluminum cup is made by filling the

space between them with water.

The outer glass cup is connected with the condenser by means

of a thistle tube which is blown into the side near the upper edge.

The distal end of the thistle tube is expanded into a tube large

enough to receive a No. 10 cork through which projects the con-

denser of the ordinary water cooled variety in common use in

chemistry work. In the outer cup is placed the solution which

forms the bath.

Acetone is used as the bath in preference to chloroform for

the sole reason that its boiling point is 56° C. (usually about 55.5° C.

or lower) while the boiling point of chloroform (Pure Chloroform

for Anesthesia) is about 58° C. and usually is slightly higher. In

fact Commercial Chloroform can not be used at all as the temper-

ature frequently exceeds 60° C. Except for the fact that Acetone

is combustible while Chloroform is not, Acetone is the better solu-

tion of the two for this purpose. However in using either Ace-

tone or Chloroform a condenser is necessary in order to keep the

bath from boiling dry and burning the tissues.

The asbestos container is built of asbestos paper and shredded

asbestos. Two asbestos paper cylinders are made using a bottle

to give the desired size and form. Cylinder No. 1 which is to con-

tain the electric bulb is placed in an horizontal position and an open-

ing the size of cylinder No. 2 is cut in the upper side near one end.

One end of cylinder No. 2 is cut to fit and placed in a vertical posi-

tion over the aperture in the side of cylinder No. 1 and the edges

sewed together. Then mixing the shredded asbestos with water it

forms a paste which is moulded over the asbestos paper cylinders

until the walls are quite thick.

34 NOTES, REVIEWS, ETC.

The bottles are now removed, the electric bulb is placed in

cylinder No. 1, and the ends closed with paste. The glass double

cup is also placed in position in the top of cylinder No. 2, and the

A, Water-jacket about aluminum cup;

A’, Water-jacket of Condenser ;

B, Aluminum Cup;

C, Inner glass cup;

D, Outer glass cup;

E, Acetone bath;

F, Thistle tube;

G, Cork;

H, Condenser;

I, Electric Bulb;

J, Switch;

K, Asbestos Container.

ELECTRIC PARAFFIN BATH

AMERICAN MICROSCOPICAL SOCIETY a5

paste carefully moulded about it. The current is now switched on

and the apparatus set aside for the asbestos to dry out.

In the drying of the asbestos care must be taken that it does

not crack as it shrinks a great deal in the process. In fact it is

better to allow the asbestos to dry without the glass cup in posi-

tion after which the container may be pared out to fit the cup. Any

patching necessary may be easily and neatly done by adding asbestos

paste.

In operation the outer cup is filled about half full of Acetone,

the top of the condensing tube is left uncorked (it being necessary

to keep it corked when the apparatus is not in use to prevent the

loss of the Acetone by evaporation), the aluminum cup with its

water jacket is placed in the inner glass cup and the water current

in the condenser and the electricity are turned on.

In order to save time, as the temperature of the apparatus is

so near the melting point of paraffin, it is better to melt the paraffin

before placing it in the apparatus. Care must be taken however

that the heat of the aluminum cup and paraffin does not break the

glass cups. This danger can be avoided by having plenty of water

in the inner cup to form the water-jacket about the aluminum cup.

Also in order that the temperature remain constant it is necessary

to carefully clean the cup containing the bath as constant boiling

causes precipitates to be formed which raise the boiling point of

the Acetone or Chloroform as the case may be.

As well as combining simplicity of mechanism, uniformity of

temperature, illumination of cup and inexpensiveness as does Mr.

Woodruft’s apparatus, this device has a lower temperature, and

greater convenience because the tissues are handled in an aluminum

cup rather than by being placed directly in the apparatus. This

allows greater ease in changing the paraffin and also makes it pos-

sible to dip the paper imbedding boxes into the paraffin containing

the tissues so that the tissues may be placed in the embedding boxes

without exposure to the air. This avoids the formation of the thin

surface film of paraffin about the tissues which so often results in

the tissues being loose in the block so that the sections fall out of

the ribbon in process of sectioning. Also the chance of breakage

is reduced to a minimum due to the breakable parts being supported

and almost surrounded by the asbestos container.

36 NOTES, REVIEWS, ETC.

The factor of safety, in one sense, is less than in Mr. Wood-

ruff’s apparatus due to the use of Acetone which forms the bath,

it being a combustible material. This danger however is negligible

as the solution is enclosed in a glass vessel and the heat is furnished

by an electric bulb.

In another sense, that of the safety of the tissues which is the

only excuse for the apparatus, the factor of safety is greater than

in Mr. Woodrufft’s machine. This is due to the fact that the con-

denser eliminates the possibility of the bath evaporating to dryness

and the consequent scorching of the tissues. Altogether the ap-

paratus is very useful and one which when once used soon becomes

a part of the permanent laboratory equipment.

Ropert W. HENDERSON.

Laboratories of Animal Biology,

The State University of Iowa.

A MACROSCOPIC METHOD OF RECONSTRUCTION

In laboratory and research work good results can often be ob-

tained by some make-shift application of materials already in stock

which would otherwise require apparatus expensive and difficult

to obtain. Such is an apparatus for the purpose of figuring to scale

objects of great irregularity of form and surface which has been

in use several months in the Animal Biology Laboratories of the

State University of Iowa.

The apparatus is really a graphing device applying the princi-

ple of the X and Y axes, and is used in conjunction with ordinary

graph paper. The materials were all found in the laboratory stock ©

room and comprise a sliding celloidin microtome, two metric rulers,

a piece of heavy wire about 18 inches long and some modelling wax.

One of the rulers, which is known as the X ruler, is fastened

in a stationary position on the frame of the microtome near to the

sliding knife carriage and parallel with its direction of motion. The

other ruler, which is known as the Y ruler, is fastened to the slid-

ing knife carriage in a horizontal position and at right angles to the

stationary or X ruler. One end of the Y ruler overrides the X

ruler, clearing it by about one-eighth of an inch.

AMERICAN MICROSCOPICAL SOCIETY 37

1. Drawing Apparatus.

a. Sliding Carriage

We 3 b. Microtome Frame

Sea c. Peep-hole

SS oy Ns x. X Ruler

hi y. Y Ruler

APPARATUS FOR RECONSTRUCTION

The heavy wire is used to make a peep-hole a foot or so above

the middle of the Y ruler and is attached to the sliding carriage.

This peep-hole is used more especially with small objects so that

the readings will all be taken from the same point thus giving great-

er accuracy. |

In attaching the rulers and wire the microtome is not injured

in the least as all the attachments are made by using screws and

clamps already a part of the machine.

In operation the object to be figured is placed on the modelling

wax under the Y ruler from which are taken the readings for the

Y axis on the graph paper. Then beginning at one end of the

object or the other the position of the Y ruler is read on the X

ruler and the corresponding line located on the graph paper. Then

38 NOTES, REVIEWS, ETC.

on this line are laid out points corresponding to certain desired points

of the object which are read on the Y ruler. When all the points

desired are located on this line the Y ruler is moved to any desired

distance, as one millimeter, along the X ruler and on the new

line corresponding to the new position of the Y ruler a new set of

points is laid out according to the readings of the Y ruler. Then

the Y ruler is again moved the desired distance and the readings

repeated and so on until on the graph paper is made a figure in dots

of the object. Now by connecting the dots by lines the figure, which

is really a graph, is complete in outline.

After the figure is complete it may be transferred to a plate

by means of carbon paper after which the completion of the figure

by shading or stippling is an easy task, and should the drawing be

spoiled in some manner another carbon tracing is easily secured.

As here described the apparatus will give a drawing of the

object as seen from a line parallel with some axis of the object

and it might be an improvement to use a stationary peep-hole above

the center of the object which would give a drawing of the object

as seen from one point.

As to accttracy each of these methods of using the peep-hole

has its advantages. The first more nearly gives a vertical view of

each point of the object which would perhaps be the ideal way for

figures. On the other hand the second method would give a draw-

ing more like a photograph. In either case the greatest accuracy

is attained by making the interval between successive readings as

small as possible. Further elimination of error may be accom-

plished by magnifying the drawing, which is done by multiplying

the reading interval by some constant number, as five, and then min-

ifying to the desired size.

Rosert W. HENDERSON.

Laboratories of Animal Biology,

The State University of Iowa.

AN EASILY ADJUSTED IMBEDDING BOX

Several types of imbedding boxes are used in histological lab-

oratories, such as paper boxes, adjustable metal right angles and

dishes of various sizes. All have their disadvantages so it seemed

AMERICAN MICROSCOPICAL SOCIETY 39

desirable to make an easily adjustable durable box, with walls thin

enough that the heat might radiate rapidly.

Fig [.

The one described here has been used several months and has

“been giving entire satisfaction. The dimensions may be changed

to suit the needs of the individual. Two rectangles of pliable sheet

tin were cut 126x20 m.m. and bent as illustrated in the top and side

projections in Fig. 1.

It is advisable to form the notches not over 2 m.m. deep with

an equal distance between them, also with as near right angles and

40 NOTES, REVIEWS, ETC.

true edges as possible. A perspective view of one adjustment is

illustrated in Fig. 2, and a top view of another adjustment in Fig. 3.

Fig. 3.

These interlocked forms after adjusting to the size needed are

placed on the usual glass plates used for imbedding. The pliability

of the tin is sufficient to hold both pieces together although it does

not inhibit opening, by bending the long ends as indicated in Fig. 3,

placing the fingers at A. and B. and bending in the direction of the

arrows.

From the Zoological Laboratory CarRLETON F. Dean.

of The Pennsylvania State College.

A DEVICE TO COUNTERACT THE EFFECT OF VIBRATION IN

PHOTO-MICROGRAPHY

In connection with the physical section of the Ansco Research

Laboratory we are frequently called upon to make high power photo-

micrographs in a building that is subject at times to considerable

vibration. Nearly all types of spring support have been tried but

were not at all satisfactory; finally these were all abandoned for

the device shown in the illustration.

A large trough (A) 9’x2’ 6’’x12” deep is attached to a founda-

tion wall by angle brackets and is not in any way connected with

the floor. The trough is completely filled with a very fine dry white

sand. It is constructed of 2’’ seasoned cypress. The bed-plates

upon which the instruments rest is 8’x2’ and is constructed of

seasoned cypress 1142” thick. The boards are tongued and glued

together. Around the edge of this bed-plate there are fixed a series

of wooden pegs 8” long. When the base-board is set down into

the sand these pegs penetrate and prevent it shifting laterally. Lev-

eling attachments are provided on the steel bed of the photo-micro-

graphic apparatus. This method has been found very satisfactory

AMERICAN MICROSCOPICAL SOCIETY 41

and even with serious vibration no blurring of a high power image

results. The particles of sand vibrate among themselves and ab-

sorb practically all of the shock. |

Binghamton, N. Y. AtFreD B. HITCHENS.

Research Laboratory.

PHOTOMICROGRAPHS OF CRYSTALLIZABLE CHEMICAL SALTS

The above is the titleof an Atlas which is a compilation

of photomicrographs made from a careful selection of many

hundreds of microscopical slides of recrystallized chemical salts.

These are made by using very weak solutions, and placing four

drops of such solution upon slides having the usual cement rings,

allowing evaporation to take place very slowly. A few of the re-

sults are presented in the accompanying figures.

The photomicrographs in this Atlas were made as one of the

steps in an endeavor to work out a method of determining the crys-

tallizable chemical salts found in evaporated saliva during a salivary

diagnosis. It was proposed to do this by means of microscopical

comparison of known salts, evaporated from known chemical so-

lutions, with the unknown salts as found in the evaporated saliva.

No attempt has been made, however, to exhaust the possible chemi-

cal combinations but rather to present typical crystalline forms of

various salts, many of which are known to exist in the body tissues

and fluids under normal and pathological conditions, thereby plac-

ing at the disposal of the student of research auxiliary help toward

an understanding of the characteristics which confront him through

the microscopic examination of fluids from the body.

In the figures shown herewith the magnification is 100 di-

ameters.

Figures 1, 2, 3, 4 and 5 are taken from the Atlas, while figure

6 is a photomicrograph made from a slide upon which was evapor-

ated the usual four drops of human saliva. Upon observing the

crystalline forms through the microscope, one having become ac-

quainted with the illustrations in the Atlas at once recognizes a

similiarity between his microscopic observation as shown in figure

6 and the photomicrograph as shown in figure 5. This is one of

the older methods of comparing the unknown with the known.

42 NOTES, REVIEWS, ETC.

Further accuracy may be gained, however, by having at one’s

command a collection of the various salts; and when a certain one

of these is suspected in the system, one may make a slide, or better,

several slides of the suspected salt. One of these slides may be

directly compared with the human material supposed to contain this

substance, by means of a comparison eyepiece, which allows one to

observe one-half of each field at the same time with one eye, thus

presenting one of the most modern methods of comparing the un-

known with the known, as shown in figure 7.

For still further identification, individual crystals may be re-

moved from the slide with a coarse hair or fine glass needle and

tested by the usual chemical methods.

The author feels that this method of studying the crystal forma-

tion would be of great value to the diagnostician, and that a course

in micro-crystallography should be made a part of medical education.

Research Publishing Co., ARTHUR W. DOUBLEDAY.

Boston, Mass.

A NEW MOUNTING MEDIUM

All of the media in present use have some defects, and it is

probable that a perfect medium will never be found, but I have en-

deavored to find one in which the most serious defects are elim-

inated.

Balsam and damar, dissolved in xylene, are ideal media for

those objects which are adapted to them. The fact that xylene boils

at a low temperature makes it easy to drive off air bubbles by very

gentle heat. But it is necessary to have all objects, mounted in

either balsam or damar, entirely free from water. There are many

objects which can not withstand dehydrating. Many delicate tis-

sues and organisms are almost destroyed from passing them through

the different strengths of alcohol necessary for mounting in balsam

or damar.

The refractive power of such media renders them unsuitable

for some objects. Objects having about the same index of refrac-

tion as these media are almost or quite invisible after they are

mounted. Again xylene balsam is never quite free from color if it

has been exposed to the light.

lives

Potassium Chloride

Ammonium Sulphate

PLATE II

er.)

; rei

mm Sia bt ”" a2

Psa el ted

Bs he

Fic. 4 #Creatinin

PLATE III

i :

i . - x *h

ace. © 3 4 4a ;

as i o :

“77 i

pe pen! {

rh, in

| Se

i Pics | ‘ } 7 ad

~~ ‘

ping Kiawah ia ;

Me i! O* Ala vf

Ay Sab eee

Pheri ss gis bie

’

car it By

eee

F nay

aa on

Lite sania

(9 S

Fic. 5. Crystals from a Known Solution

Fic. 6. Crystals from a Human Secretion

PLATE LV

a ee 2 . . : 7 a

® . , - _—— I SS Oa - piensa

Fic. 7. Left half of photomicrograph presents crystals from

Human Secretion: right half from a Known Solution.

Fic. 8. Muscles in neck of Tussock Moth (see page 48).

PLATE V

AMERICAN MICROSCOPICAL SOCIETY 43

With any of the media containing gelatine it is impossible to

get rid of air bubbles. They cannot be driven off by heat without

destroying the qualities of the medium. There are the same objec-

tions to media containing acacia. Neither of these media is miscible

with alcohol.

A medium different from any of the ordinary kinds is needed

for mounting some objects. The qualities desirable in such a new

medium may be classified as follows: 1, It should be miscible with

both alcohol and water; 2, It should be free from color; 3, It

should be free from air bubbles, or nearly so; 4, It should have

a refractive index differing from that of balsam etc.; 5, Gentle heat

should carry off the bubbles, if they are present. 6, Gentle heat

should not change its nature in any way; 7, It should harden spon-

taneously, or by the application of gentle heat.

I think I have discovered a medium that has all of the above

mentioned good qualities except the last. It is made from con-

fectioners glucose. This, as found in the market, is a gelatinous

substance, very transparent and free from color. Probably in the

process of manufacture it is strained through cotton cloth. At any

rate there are some fibers in it. To remove these fibers it is neces-

sary to dilute the glucose with water until it will pass through a

paper filter. The water should be driven off by heat over a water

bath until the glucose is considerably thicker than xylene-balsam.

Then alcohol should be added until it is of the same consistency as

xylene-balsam, or a little thinner. The glucose is not perfectly

soluble in strong alcohol but is soluble in alcohol and water com-

bined. If adding the alcohol produces a milky appearance it will

be necessary to add a little water to make a clear solution, then the

medium is ready for use.

Mounting with the medium is performed in the same way as

with xylene-balsam except that it is not necessary to dehydrate ob-

jects. Gentle heat applied to the slide will not only drive out the

air bubbles but it will also harden the glucose around the cover

glass so as to hold it firmly. If this condition would persist, it

would be very satisfactory; but unfortunately the glucose is hygro-

scopic and the moisture taken from the air will soften it so that

the cover glass can be removed easily. To avoid this a ring of

Ad NOTES, REVIEWS, ETC.

enamel can be spun around the edges of the cover glass easily and

quickly, and then the preparation is a permanent one. Only one

layer of enamel is necessary. I think any kind, or color, of enamel

can be successfully used. I have used two different kinds and

both seem satisfactory.

A precaution should be taken that the solution is not so thick

that it will harden at the edge of the cover before the bubbles are —

all driven off.

I have been using this medium, and method of mounting, for

about six months. The objects which I have mounted seem to

be in perfect condition at present. If time and changing temper-

ature should loosen the cover glasses I think that my method might

be changed with advantage by putting a ring of gold size around

first, and following this by a second coat, or a coat of enamel.

Lewiston, Maine. C. E. Norton, M. D.

A METHOD FOR CUTTING PLANORBIS EGGS

The eggs of Planorbis, a common pond snail, are found in

clusters held together by tough enclosing membranes which con-

tain a considerable amount of jelly. Surrounded by the jelly are

perhaps a couple of dozen capsules, each filled with a yellow albu-

men mass in which the egg develops. These membranes and the

albumen make it very difficult to section the entire cluster, so that

it has usually been necessary to remove the eggs from the albumen

and jelly. Holmes (Journal of Morph., Vol. 16, 1900) recom-

mended for the process that they be teased out in physiological

salt solution to which a trace of picric acid had been added.

The difficulties, however, in the process of staining and run-

ning through the alcohols of so small a number of eggs which are

microscopic in size led to further experiments on cutting the eggs

within the jelly mass. The procedure which was developed for

this purpose has proven fairly successful, although certain precau-

tions are necessary.

For fixing the eggs, Kleinenberg’s picro-sulphuric has been

chiefly employed. Owing to the tough membranes a longer time

than is ordinarily given to mollusc eggs is necessary for the proper

fixation; about one hour has proven a satisfactory time. The ad-

AMERICAN MICROSCOPICAL SOCIETY . 45

dition of a small percentage of acetic acid is perhaps an advan-

tage. It is well also to puncture the enclosing membranes to insure

a quick penetration. Especial care must be paid to this matter of

fixation, for the tough membranes tend to interfere with the process.

The fixitive should be washed out of the eggs in 70% alcohol

to which lithium carbonate has been added. If the dish contain-

ing the eggs is kept in a warm place and the alcohol changed sev-

eral times, forty-five minutes to an hour will be sufficient for the

washing. The egg masses should then be transferred to 80% al-

cohol for a few minutes. To clear the egg masses they should be

placed in creosote directly from the 80% alcohol. The success

of this method depends to a large extent upon avoiding the use

of higher grade alcohols and xylol; (even the 80% should be used

for only a few minutes.)

As soon as the eggs have cleared in the creosote (which takes

place in less than half an hour) they must be removed and rinsed

for a short time in chloroform. If they remain long in the creosote

they are hardened and toughened. After running through a chloro-

form-paraffin mixture, the eggs are infiltrated for about one hour

in fairly strong rubber paraffin (J. B. Johnston, Journal Appl.

Miscros. Vol. VI). Care is necessary there that the temperature

does not rise higher than actually necessary. The entire process

from fixation to embedding should all be concluded the same day

if satisfactory results are to be expected.

Since the jelly in the sections stain readily, the best stain

so far tried with these sections is thionin which is selective between

the eggs and the jelly. The sections are stained in .5% aqueous

solution of thionin for twelve to eighteen hours, and then differen-

tiated in a weak solution of Orange G in 95% alcohol, quickly de-

hydrated and cleared, and then mounted as usual. Thionin, of |

course, has the objection that it is not lasting; otherwise its use

proves quite satisfactory. For more lasting preparation, the usual

staining methods may be used, but the fact that with them the jelly

stains readily renders their use less desirable.

Wabash College, A. RICHARDS.

Crawfordsville, Ind.

46 NOTES, REVIEWS, ETC.

METHODS REPORTED FROM THE ZOOLOGICAL LABORATORY, UNIVERSITY

OF WISCONSIN

Potash Clearing Method.—The potash clearing method is of

great value in demonstrating skeletal structures without the removal

of overlying tissues, and also in showing the relations of unossified

bone as in the tarsus of amphibians.

The animal or part should be skinned and while fresh Ae

into a 1% solution of KOH for about 12-24 hrs., or until internal

structures are visible. The tissue is then removed to pure glycerine

where it may be kept indefinitely. Animals as large as adult bull-

frogs have been successfully cleared in this laboratory.

Substitute for Spalteholtz Clearing Method.—The expense con-

nected with the Spalteholtz clearing method makes it prohibitive

for general class use. A good substitute consists in the use of ben-

zaldehyde as the clearing agent. The tissue after injection or other

treatment is dehydrated in alcohol as far as 95%, and then re-

moved to benzaldehyde and the container at once sealed. Benzalde-

hyde is very unstable, oxidizing in the air to form benzoic acid,

hence it must not be exposed to the air. Specimens cleared in it

must be carefully sealed.

Clearing and Mounting Hydra.—Some difficulty is encountered

in clearing Hydra in xylol after dehydration in alcohol since the

tissue becomes too brittle to be easily handled. This is especially

true in making mounts of Hydra to show the developed ovaries

and spermaries, which often drop off during the process. We

have found that with rapid dehydration in the alcohols, followed

by clearing in wintergreen oil this excess brittleness may be avoided.

Mounts of Hydra bearing six or more spermaries and a well de-

veloped ovary have been made without difficulty, mounting in bal-

sam in the usual way.

Injection of Semicircular Canals.—In using the semicircular

canals of the shark for demonstration in elementary courses, where

the entire head is immersed in a specimen jar, we find that stu-

dents have much difficulty in making out the canals and ampulle.

By injecting these with India ink, introduced with a fine pipette

the structures are made plainly visible. The ink will remain in

AMERICAN MICROSCOPICAL SOCIETY 47

the canals if the specimen is given ordinary care after sealing in

a specimen jar. |

Ciliated Epithelium for Histological Study.—Students of histol-

ogy often find it hard to obtain permanent slides of ciliated epithel-

ium which show the separate cilia satisfactorily. There is a great

tendency for these structures to matt down in a confused mass

during fixation. I have had very good success using the sheath of

the crystalline style of the fresh water mussel (Anodonta or Unio)

and fixing in Bouin’s fluid. After opening the animal from the

right side cut through the stomach wall and, beginning at the opening

of the intestine, slit it open for several centimeters. This will re-

veal the two typhlosoles with the style sac between them. Care-

fully free these structures, for a short distance, from the under-

lying tissues and place in a vial with the ciliated surface upper-

most. Keeping the tissues as flat as possible, without pressing

the ciliated side, run in enough of the Bouin’s fluid to cover fully,

and fix 12-24 hrs. Run up through the alcohols as usual. 1 find

that sections cut 5 and stained in iron hematoxylin, and counter-

stained with any acid carmine stain give the best results.

Preparation of Chick Embryos for Demonstration.—Students

show much interest in the living chick embryo, especially when its

heart is visible. The following method may be used to prepare

them so that they may be watched for several days. The egg is

taken from the incubator at the stage desired and placed in a bed

of warm cotton. With a small compass a circle of the desired size

is drawn on the shell, and this is then scored with a safety razor

blade. With a pair of small scissors follow this scored line cutting

just deeply enough to get through the shell. Lift off the circular

piece of shell, drain off a little of the thinner albumen, and place over

the opening a thin sheet of celloidin, cementing it fast with a solu-

tion of celloidin in ether.

This dries very rapidly, and as soon as the edge of the mem-

brane is fast turn the egg over so that the embryo will float up

against the shell on the opposite side of the egg until the membrane

finishes drying. Keep the egg in a small incubator with a glass

top, and provide abundant moisture to prevent the embryo from

drying against the membrane.

48 NOTES, REVIEWS, ETC.

The sheet of celloidin is easily made by pouring a thin layer

of celloidin in alcohol and ether on a clean glass plate.

Zoological Laboratory, TuHurRLtow C. NELSON.

Univ. of Wisc.

EVIDENCE AS TO THE NUMBER OF SEGMENTS IN THE HEAD OF INSECTS

Students of insects estimate the number of somites entering

into the insect head variously from one to nine. It appears to the

writer that the number of muscle units in the mass of neck muscles

may have some bearing on this question. The accompanying section

(Plate V, figure 8) shows the condition in the neck of the tus-

sock moth. Here we find five pairs of superimposed muscle bun-

dles extending into the head. This fact perhaps strengthens the

view that the five pairs of modified appendages about the head

stand for so many segments. This muscular arrangement is quite

generally found in insects.

E. W. Roperrws.

ANIMAL MICROLOGY

This well known book, now also well used for ten years, has

recently been reissued in an enlarged second edition. The new

volume represents a 20% increase over the original. The plan of

the first edition has been maintained. This consists of a series of

chapters dealing with the steps necessary in handling various kinds

of animal materials; the reason for taking these steps in the way

suggested; the points where difficulties are most likely to be en-

countered and the way to avoid these; and thus the means of dis-

covering and remedying defective results.

In appendices are given an elementary treatment of the mic-

roscope and the principles of its making and use; a record of the

formulz for the making of the most used reagents, and suggestions

for using them; a table of the tissues and organs of animals, and

the most accepted method of killing, fixing, hardening, sectioning,

staining, and other technic if any; a brief description of the meth-

ods peculiarly suitable to various groups of animals; and finally

tables of weights and measures, and equivalents.

AMERICAN MICROSCOPICAL SOCIETY 49

While the plan has not been changed, changes have been made

thruout. New items have been added, many sections have been

wholly recast, and two new chapters have been inserted. In these

various ways has the essential progress of the intervening years been

recorded. The new chapters are, “Some Cytological Methods,” and

“Drawing.” Each of these adds distinctly to the value of the book

to the student and teacher. The chapter on drawing fills a very

real need in our histological laboratories. The book is sure to con-

tinue one of the most useful in existence to teachers and students

in zoological laboratories.

Animal Micrology; Practical Exercises in Zoological Micro-Technique. By M.

F. Guyer. University of Chicago Press. 1917. Price, $2.00 net.

AN INTRODUCTION TO NEUROLOGY

Under this title Professor Herrick has issued a book designed

to enable the student to organize his conceptions of the regional

differentiation of the nervous system in man, and to relate to this

the functional diversity, and to do this from the beginning. In

other words this treatise summarizes the facts and relations which

will satisfactorily introduce the reader to the necessary foundations

of neurology without all the complexity of detail presented in the

elaborate manuals of the subject. This fact makes the book pecul-

iarly valuable to students of medicine, psychology, hygiene especially

of nervous processes, education, sociology, general zoology, phys-

iology,—not only as furnishing an undergraduate course in neurol-

ogy preliminary and introductory to these, but as an aid and sup-

plement to such undergraduate studies and to the general student.

In the opinion of the reviewer the author has done a real service

to the teacher and student who needs a simplified yet consistent

statement of the facts and principles of neurology.

Chapters I to VII discuss the more general neurological top-

ics,—as nervous functions in the large, the neuron, reflex circuits,

receptors and effectors, the anatomy and functions of the nervous

apparatus. Chapters VIII to XVIII present an analysis of the

chief subdivisions of the nervous system, including the organs of

reception, and the functions of these various parts. The conclud-

50 NOTES, REVIEWS, ETC.

ing chapters, XIX to XXI, discuss in greater detail the structure,

the localization of function, and the evolution of the cerebral cortex.

A very elaborate index, in which many terms are defined as

well as paged, concludes the book. This index is one of the most

adequate and effective which the author recalls in a book of this

size.

An Introduction to Neurology, by C. Judson Herrick. 354 pages; illustrated.

W. B. Saunders Co., Philadelphia, 1916. Price $1.75, net.

NOTEWORTHY

GALLOWAY—Zoology. A Textbook for

Universities, Colleges and Normal

Schools. By T. W. Galloway, Ph.D.,

Beloit College. 3rd Edition Revised and

Enlarged. 255 Illustrations. Cloth $2.00

Postpaid.

GALLOWAY—Elementary Zoology. For

Secondary Educational Institutions. 160

Illustrations. Cloth $1.25 Postpaid.

KINGSLEY—The Comparative Anatomy

of Vertebrates. By J. S. Kingsley, Unt-

versity of Illinois. 346 Illustrations.

Cloth $2.25 Postpaid.

PATTEN—The Evolution of the Verte-

brates and Their Kin. By William Pat-

ten, Ph.D., Dartmouth College. 309 Il-

lustrations. Cloth $4.50 Postpaid.

BOOKS

GAGER—Fundamentals of Botany. By C.

Stuart Gager, Brooklyn Botanic Garden.

435 Illustrations. Flexible Cloth $1.50

Postpaid.

LEWIS and STOHR—Textbook of His-

tology. 2d Edition. 495 Illustrations.

Cloth, $3.50 Postpaid.

McMURRICH—Development of the Hu-

man Body. A Textbook of Embryology.

By J. P. McMurrich, University of Tor-

onto. 5th Edition. 287 Illustrations.

Cloth $2.50 Postpaid.

HAMAKER—Principles of Biology. By

J.’ I. Hamaker, Randolph-Macon Wo-

man’s College. 267 Illustrations. Cloth

$1.50 Postpaid.

P. BLAKISTON’S SON & CO.

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VOLUME XXXVI No. 2

TRANSACTIONS

OF THE

AMERICAN

MICROSCOPICAL

SOCIETY

\ PUBLISHED QUARTERLY BY THE

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} DECATUR, ILLINOIS

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The Secretary will consider it a favor if members will notafy

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TRANSACTIONS

OF THE

American Microscopical

Society

ORGANIZED 1878 INCORPORATED 1891

PUBLISHED QUARTERLY

BY THE SOCIETY

EDITED BY THE SECRETARY

VOLUME XXXVI

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TABLE OF CONTENTS

FOR VOLUME XXXVI, Number 2, April, 1917

A Monostome Lung Fluke from the Painted Terrapin, Chrysemys Mar-

ginata Agassiz, with Plates VI and VII, by F. D. Barker and

Susdtine Parsons, sii os eee Fee paw M Rie wp Als ae w Ag bb kW lols Wie Mimi gree Re

Enchytreide (Oligocheta) from the Rocky Mountain Region, by Paul

BVI ee Ee CRE CT teks ge aa llprana ob fee ie Onis ie et ace ener

The Morphology, Structure, and Development of Hydractinia Polyclina,

with Plates VIII and IX, by J. A. Place..........ce cece eeeeeeoees

Another Cestode from the Young Cat, by J. E. Ackert and A. A. Grant

Notes and Reviews: Entomological Notes, by Paul S. Welch; Notes on

Oligocheta, by Paul S. Welch; Parasites in the Mouth in Cases of

Pyorrhea; Factors Influencing the Sporangial Characters of Myce-

tozoa; Factors Controlling the Rate of Regeneration; Head and

Mouth Parts of Diptera; Early Castration of the Vertebrate Em-

bryo; Segregation and Recombination of Chromosomes ; Laying Cy-

cles in Birds; The Pineal Gland and Pigmentation; Cooperative

Technic; Fixation in Mammalian Chromosomes; Quince Jelly as a

Culture Medium for Euglena; “Vital Dyes” in Teleosts; Acid Col-

loidal Dyes in Blood and Tissue Cells; Genetics and Eugenics (W.

ATH: iT) 0 CON el eR AMY SOME, ROCHA H IL EY ELIANA TRS he

Spencer-Tolles Fund Report for 1916..........seee eee ceee cere eeneeees

Necrology: C. E. Hanaman, with portrait...........sseeeeeeeeeeeeceees

(This Number was issued on July 12, 1917.)

TRANSACTIONS

American Microscopical Society

(Published in Quarterly Installments)

eee

Vol. XXXVI APRIL, 1917 No. 2

A MONOSTOME LUNG FLUKE FROM THE PAINTED

TERRAPIN, CHRYSEMYS MARGINATA AGASSIZ

By FRANKLIN D. Barker and SUSANNE Parsons

With plates VI and VII

SYNOPSIS

SIMS MMUACRIGAL LUST SUS Vibe 4's Wr be oon ahead s Oe RE Pe aa TU Ra ST AN 55

PRMCRIMREIER NN ale Ce Sa Wi ee ald Waid Seo ee kee de Sed eR te FORA o 56

DRC MOM UTOT ELOMACOME: a, sk yb ulec A wp Bie staccato uot es 57

MEIC LRR yee EY sc leealy WINE acho heli'§ Wakes AOA WIE ME Ge aie Oe Le 62

Rae CMEEME icy hal th © wi nldn yom aie axa ve Fotis Te a RRL ie ake clara 65

PIA PMESALIOIM OS) L LACES (55 ae alk wa og Aik Sa coed UP Meee eae MEG eth 65

INTRODUCTION

The trematode here described was first found in August, 1911,

by the senior author in the lungs of the painted terrapin, Chrysemys

-marginata Agassiz, taken from Lake Emily, a small lake near St.

Peter, Minnesota. A preliminary study of the parasite was made

at that time, and subsequently this material was turned over in

September, 1913, to the junior author, who in collaboration with

_ the senior author has made a detailed study of, the parasite.

Studies from the Zoological Laboratory, The University of Nebraska, No. 116

56 FRANKLIN D. BARKER AND SUSANNE PARSONS

‘TECHNIQUE

During August and September of the summer of 1911, seven

turtles of the species Chrysemys marginata Agassiz, were secured

from Lake Emily. Upon examination of the lungs it was found

that two of these turtles were uninfected, while of the remaining

five, one yielded 13 trematodes from both lungs, one yielded 1 from

one lung, a third and fourth 5 and 3 respectively, and the fifth yield-

ed 3 from one lung. Later, during the months of November and

December of the same year, sixteen more turtles from the same

lake were examined with the result that only one showed a par-

asitic infection of the lungs and only one trematode was obtained.

It may be stated, however, that these turtles had been kept in con-

finement from one to two months, which together with the time

of year, may account for the small percentage of infection.

Two years later, during the fall of 1913, seven turtles of the

same species were obtained from the Fairport U. S. Biological Sta-

tion, near Fairport, Iowa, on the Mississippi River. The lungs of

these seven turtles were examined and it was found that two, one

male and one female, were uninfected, and of the remaining five,

two had both lungs infected, both being male animals and furnished

three and six trematodes respectively. Of the three having one

lung infected, one female and one male furnished one trematode

each, while the third, a female yielded thirteen trematodes, four

of which were together in one pocket of the lung. The minimum

number of trematodes obtained from a single turtle was one, and

the maximum number thirteen. The three male turtles furnished

a total of ten trematodes, and the two female turtles furnished a

total of sixteen trematodes, showing the females almost three times

as heavily infected as the males. Possibly different habits or food

eaten may have a bearing on this difference in the degree of infec-

tion. This same species of turtle secured from the creeks and

ponds near Lincoln, Nebraska, has never been found to harbor

these trematodes. |

To kill the trematodes, the specimens were compressed between

slides, thus keeping the worm in a fairly well distended condition,

A MONOSTOME LUNG FLUKE 57

and hot solutions of alcoholic-corrosive-acetic, Zenker’s, Bouin’s or

lacto-phenol, were run between the slides. The toto-mounts and

sections were stained in borax carmine, Le Brun’s, and Delafield’s

hematoxylin.

For the study of the eggs, worms which had been killed in

lacto-phenol or Bouin’s solution were teased up in those solutions

and the eggs isolated, thus insuring the least shrinkage and distor-

tion of the eggs.

MorPHOLOGY

The description given is based on the comparative study of

fifteen specimens mounted in toto, and ten series of sections cut

longitudinally, transversely, and sagittally.

The worms were, when living, a reddish brown, with a thick

fat elongated body which curled ventrad, along the long axis. In

one specimen the intestinal ceca were observed to be a bright blood-

red color, indicating that the worms feed upon blood in the lung.

The uterine coils at each side of the body in all specimens were

dark, and the longitudinal, straight tubes of the uterus were espe-

cially black, due to the presence of dark granules. All specimens

were characterized by a marked sluggishness.

The worms vary in length from 12.24 mm. to 4.899 mm., the

mode being 9.5 mm. for 43 per cent. The width at the widest

part of the body varies between a maximum of 2.65 mm. and a min-

imum of 1.57 mm. The body is widest at the level of the second

anterior fifth, and tapers gradually toward the posterior end, which

is rounded. From this same level the body tapers anteriorly more

rapidly and the extreme anterior end is decidedly pointed (Plate VI,

figs. 2 and 6). With the exception of the two ends the sides of

the body are almost parallel and the dorso-ventral flattening is

marked. Strong circular bands of muscle fibers run around the

body at regular intervals, giving it a segmented appearance when

contracted. The oral sucker, the only one present, is sub-terminal

in position and is a globular fleshy structure, relatively small but

distinct. Its musculature is conspicuously weak, it being composed

mostly of parenchyma tissue. (Plate VII, fig. 3.) The length of

58 FRANKLIN D. BARKER AND SUSANNE PARSONS

the oral sucker varies between a maximum of 0.314 mm. and a min-

imum of 0.204 mm., the mode being 0.245 mm. for 56 per cent.

The width of the sucker varies from a maximum of 0.455 mm. to

a minimum of 0.314 mm., the mode being 0.435 mm. for 54 per cent.

Just posterior and very close to the oral sucker is the pharynx,

a very muscular and almost globular structure, formed by two bean

shaped halves. The pharynx varies in length from a maximum

of 0.345 mm. to a minimum of 0.219 mm., the mode being 0.314 mm.

for 44 per cent. The width of the pharynx varies between a max-

‘mum of 0.314 and a minimum of 0.219 mm., the mode being 0.282

mm. for 44 per cent.

The pharynx is followed by a very short, wide esophagus. Its

modal length is 0.047 mm. and its modal with 0.143 mm. It was

impossible to determine its length and width in all specimens, due

to a contracted condition but these measurements were fairly con-

stant.

Immediately posterior to the esophagus, the digestive tube

branches into two wide intestinal ceca, which pass posteriorly as

straight unbranched tubes to within 0.282 mm. of the posterior

end of the body where they end blindly. The ceca occupy from

one-fifth to more than one-half of the width of the body, leaving

only a small space between them in the median line of the body.

In all the specimens examined clots of blood or mucous from the

lungs of the turtle were found in the intestinal ceca.

Female genitals: A well-defined ovoidal ovary lies in a dorsal

position slightly posterior to the bifurcation of the intestine and to

the right or left of the median line. Its length varies from 0.392 mm,

to 0.518 mm. the mode being 0.44 mm. for 57 per cent. Its width

varies from 0.314 mm. to 0.455 mm., the mode being 0.35 mm.

for 70 per cent. Its margin is smooth. A small ciliated oviduct

leads caudad from the posterior margin of the ovary and after mak-

ing several close coils joins the vitelline duct. The union of these

two ducts marks the beginning of the uterus which after making

several compact turns dorsal to the testis and caudad to the ovary

turns to the left or right just caudad to and across the base of the

ovary and passing posteriorly enters a well defined and compact

shell gland about one-fourth to one-half the size of the ovary and

A MONOSTOME LUNG FLUKE 59

situated obliquely caudad to the ovary (Plate VI, fig. 4.) The

uterus may be divided into two parts, the coiled lateral portion,

extending from the level of the ovary to the posterior end, lying

along the sides of the body and extending mesad over the intestinal

ceca and the arms of the vitellarium and the straight median por-

tion extending from the posterior end to the genital pore. After

emerging from the shell gland the uterus takes the following course

in loose coils dorsal to the vitellarium, posteriorly to the level of

the anterior third of the body and just caudad to the seminal vesicle

then in a straight course transversely and obliquely ascending across

the body dorsal to the intestinal cecum to the level of the ovary,

then in descending coils along that side of the body to the posterior

end obliquely across to the opposite side of the body and in ascend-

ing lateral coils to the level of the bifurcation of the intestine where

it turns mesad and in a straight course passes to the posterior end

of the body in the median plane then turning back on itself ascends

in a straight course parallel and ventral to the descending limb to

the common genital pore just ventral to the posterior margin of

the oral sucker.

The lateral uterine coils are less numerous and smaller in the

younger worms while the median straight tubes are larger than

in the older worms in which the lateral coils are numerous and

compact and the median tubes narrow. The lateral coils are crowd-

ed with developing eggs while the median straight portions con-

tain comparatively few dark colored eggs. Masses of sperm cells

are found all along the course of the uterus but are most abundant

in the terminal portions, near the ovary and the genital pore.

The eggs are very abundant in the coiled lateral portions of

the uterus but few are found in the median portion. Those in the

median portion are fully mature and are dark brown in color. They

are egg-shaped, one end being slightly more pointed than the other.

The shell is thin and no operculum could be found. They vary

in length from 0.045 mm. to 0.065 and in width from 0.022 mm. to

0.030 mm. The largest eggs are found in the smaller worms. (Plate

WaPo die.) 5.)

The vitellarium is a voluminous “U”’ shape organ with its apex

or closed end at the level of the anterior fourth or sixth of the body

60 FRANKLIN D. BARKER AND SUSANNE PARSONS

just posterior to the ovary. The two arms are lateral in position

and lie dorsal to the uterine coils and the intestinal ceca. They are

somewhat coiled and extend to the posterior seventh or ninth of

the body. From the posterior half of each arm a vitelline duct

arises and passing ventrad around the outside of each intestinal

ceca passes cephalad under or mesad to the ceca to the level of the

apex of the vitellarium where they turn mesad and unite to form

a small yolk reservoir posterior to the ovary and just anterior to

the apex of the vitellarium. A short vitelline duct passes ante-

riorly in the median line and joins the oviduct just caudad to the

ovary. Neither a Laurer’s canal nor a seminal receptacle is present.

Male genitals: In the older, larger worms, 6 to 12 mm. long,

a testis is not discernable but in the smaller worms, 3.5 to 6 mm.

long, a single spherical or ovoidal testis with smooth margin, one-

half to one-fourth as large as the ovary, is present, lying ventral

to and partly under or just posterior to the ovary. The testis is

often obscured by the first compact coils of the uterus which lie

dorsal to it. A small duct, the vas deferens, leads dorsad from the

anterior end of the testis and enters a well defined ovoidal and

comparatively large prostate gland which lies dorsal to the testis.

(Plate VII, fig. 4.) The vas deferens emerges from the prostate

gland and passing caudad and ventral to the transverse portion of

the vitellarium enters the seminal vesicle. The seminal vesicle is

a prominent, wide, fish-hook shaped structure, extending from the

genital pore in an undulating median course to the posterior mar-

gin of the anterior third or fourth of the body. The short arm of

the vesicle lies dorsal to the long arm. The seminal vesicle merges

without demarcation with the lumen of an eversible non-muscular

cirrus formed by an introvert of the outer body wall of the worm.

The cirrus evaginates thru a common genital pore which lies ventral

and slightly lateral to the median line at the outer margin of the

oral sucker. (Plate VI, fig. 1.)

We seem to have in this trematode a protandric condition in

which both female and male genitals are fully developed in the

younger (smaller) worms but the testis gradually atrophies and

entirely disappears in the older (larger) worms. Coincident with

this disappearance of the testis, the seminal vesicle increases in size

A MONOSTOME LUNG FLUKE 61

and becomes distended with sperm cells and the uterine coils and

the number of eggs increase enormously in volume.

Looss (’02:676) describes a similar condition in the genus

Microscaphidium, except that the ovary atrophies also, as follows:

“Die einzige Frage, auf die ich zur Zeit noch keine Antwort zu

geben vermag, ist die, warum die Angehorigen der Gattung Micro-

scaphidium allein von allen mit ihnen zusammen in demselben

Wirthe lebenden Parasiten im Laufe von Monaten und unter allem

Anscheine nach giinstigen Verhaltnissen nicht zur Production von

Eiern schreiten. In dieser Zeit werden sie nattirlich alter, und

eskommt hierbei nun thatsachlich vor, dass die Keimdriisen ganz

oder theilweise atrophiren, ehe sie tiberhaupt in Function getreten

sind. Unter den im Monat October gesammelten grossten Exem-

plaren von M. reticulare und aberrans fanden sich versehiedene,

die in ihren Bewegungen und ihrem ganzen wtbrigen Verhalten

genau so lebhaft waren wie ihre Genossen, deren Hoden aber da-

durch auffelen, dass in ihnen eine grdssere Anzahl orange-bis-

ockergelber Tropfchen und Kugeln einer speckigen, starker lichtb-

rechenden Substanz enthalten waren. Nachdem die Thiere in Al-

kohol conservirt, gefarbt und eingeschlossen waren, zeigte sich, dass

mit der Zunahme dieser Kugeln eine auffallige Abnahme der zelligen

Elemente der Hoden Hand in Hand ging. Bei einem Individuum

bei dem dieser Degenerationsprocess der Hoden am _ weitesten

fortgeschritten war, fanden sich im hintern Hoden tberhaupt keine

gefarbten zelligen Elemente mehr, im vordern nur noch enige wenige,

in beiden dagegen eine grosse Menge der erwahrten Kugeln und

Schollen. Die Samenblase zeigte vollkommen das Aussehen,

' welches sie darbietet, ehe sie in Function tritt. Im Keimstock fanden

sich zwischen den normalen jugendlichen Eizellen hier und da gelbe,

kriimlige Massen, die in den Hoden die Anfangsstadien in der

Bildung der gelben Kugeln darstellen.”

Self fertizilation is of common occurrence if not the only method

as shown by several specimens in which the cirrus is found extruded

and inserted into the vagina or metraterm of the same individual

(Plate VII, fig. 3.).

Excretory system: The main system consists of two lateral

canals first noticeable in cross sections, about the level of the

62 FRANKLIN D. BARKER AND SUSANNE PARSONS

esophagus running posteriorly, just mesad and ventral to the intes-

tinal ceca, for two-thirds the length of the body, where they empty

into a large median irregular excretory bladder or reservoir. The

reservoir is dorsal in position and extends from the middle of the

body posteriorly to the end of the intestinal ceca where it ends

blindly. The bladder opens through the excretory pore in the

median line on the dorsal surface just anterior to the posterior end

of the body (Plate VI, fig. 3.). The nervous system is of the

typical trematode type consisting of two ganglia one on each side of

the pharynx and connected by a dorsal supra-pharyngeal commis-

ure. Two lateral nerve trunks, ventral in position, extend from the

ganglia to the posterior end of the body. (Plate VII, figs. 2, 5, 6.)

TAXONOMY

Several systematists, Looss (’02:254), Lithe (’09:24), Kossack

(711:491), Skrjabin (13:90) have given schemes of classification

for the monostome trematodes. Of the families suggested by these

authors, the characters of this trematode from the lung of the

terrapin conform most nearly to characters of the family Cyclo-

coelidae of Kossack ('11:496) which he describes as follows:

“Digenetische Trematoden, von grossem bis mittelgrossem Korper,

ohne Mundsaugnapf. Bisweilen ist ungefahr an der Grenze des

ersten Korperdrittels ein Bauchsaugnapf vorhanden. Haut beim

konservierten Tiere mit zahllosen Griibchen versehen. Mundoffnung

endstandig oder subterminal. Der verschieden, aber nie erheblich

lange Oesophagus besitzt einen muskulésen Pharynx von wechselnd-

er Grdsse, dessen relatiy Entfernung von der Mundoffnung

schwankt. Darmschenkel einfach oder an der Innenseite mit kurzen

Blindsacken versehen, im Hinterende des Korpers bogenformig in-

einander tibergehend. Excretionsporus kurz vor dem Korperende auf

dem Riicken. Excretionsblase zwischen Darmbogen und dem Kor-

perende. Genitalporus median, nicht weit hinter der Mundoffnung.

Copulationsorgane vorhanden, aber wenig kraftig entwickelt.

Samenblase im Cirrusbeutel. Dotterstécke zwischen Korperwand

und den Darmschenkeln gelegen, letztere bisweilen umspannend

und in diesem Falle im Hinterende wie sie kontinuierlich ineinander

A MONOSTOME LUNG FLUKE 63

tbergehend. Geschlechtsdrtisen zwischen den Darmschenkeln. Die

beiden Hoden einfach oder gelappt schrag zueinander gelagert.

Keimstock immer ganzrandig. Laurer’scher Kanal und Receptacu-

lum seminis fehlen. Uterus sehr stark entwickelt, mit regelmas-

sigen quer gerichteten Schlingen den Korper fast vom Hinterende

bis zur Darmgabelung erfiillend. Eier sehr zahlreich, von nicht ein-

heitlichem Aussehen, ohne Polfaden. Sie nehmen im Laufe der

Entwicklung an Grdésse zu und enthalten schon im Uterus das ausge-

bildete Miracidium, dessen doppelter Augenfleck ihnen ein charak-

teristisches Aussehen verleiht.” |

It is evident, however, that the. trematode here described differs

from the family Cyclocoelidae in several essential characters such

as the presence of an oral sucker, the anterior position of the ovary,

the single testis, the form and position of the vitellarium and the

extent and form of the uterus. But rather than resort to the ques-

tionable procedure of creating a new family or even a sub-family

based on a single new genus we prefer for the present to enlarge

the characters of the family Cyclocoelidae as given by Kossack so

that it may contain this new genus. However, a comparative study

of the genera in the family Cyclocoelidae as given by Kossack ’11

and Skrjabin ’13 clearly shows that none of them will accommodate

this trematode, this necessitates the creation of a new genus which

we in a previous paper, Barker and Parsons (’14:193) have desig-

nated as dorchis (without testis). This new species of lung fluke

from the terrapin has previously been designated as Aorchis extensus

(voluminous uterus) and becomes the type species of the genus

Aorchis having the following characters.

Genus Aorchis Barker and Parsons.

Body medium to large size, slightly tapering toward the anterior

and posterior ends, posterior end rounded. Oral sucker small,

weak but distinct. Mouth opening terminal. Pharynx strongly

muscular, without pockets. Esophagus short. Intestine composed

of two simple blind sacs, not uniting at the posterior end. Genital

pore not prominent, ventral to the pharynx, close to oral sucker.

Ovary anterior between intestinal ceca. Shell gland compact, pos-

terior to, and smaller than ovary. Uterus made up of coils which

fill the body lateral to and overlap the intestinal ceca, extending from

64 FRANKLIN D. BARKER AND SUSANNE PARSONS

level of the ovary to posterior end of the body and two straight and

parallel uterine tubes which pass anteriorly up the median axis of

the body between the intestinal ceca. Vitelline gland a voluminous,

coarse, compact, U shaped mass posterior to ovary and dorsal to the

intestine, with the closed portion at anterior end. Protandrous.

Testis absent, or atrophied in old worms. A single ovoidal testis

present in young worms, anterior, caudad to ovary. Prostate gland

near testis, seminal vesicle, a large tubular structure extending from

genital pore to the level of the second anterior fifth of the body.

Protrusible non-muscular cirrus present. Laurer’s canal and

seminal receptacle absent. Eggs without lids. Excretory pore

posterior dorsal.

Type species: Aorchis extensus Barker and Parsons.

Habitat—Lungs of painted terrapin Chrysemys marginata

Agassiz.

Distribution—Illinois, Iowa and Minnesota, U. S. A.

We wish to acknowledge our indebtedness to Doctor Robert E.

Coker, formerly Director of the U. S. Biological Station, Fairport,

Iowa, for assisting us in securing material for this study and in

granting both authors the use and facilities of the Station Labora-

tory during the summers of 1913 and 1915.

PAPERS CITED

Barker, F. D. and Parsons, SUSANNE

1914. A new species of Monostome from the Painted Terrapin, Chry-

semys marginata. Zool. Anz. V. 45. No. 5. pp. 193-194.

Kossack, WILLY

1911. Ueber Monostomiden. Zool. Jahrb. Syst., V. 31, pp. 490-590.

Looss, A. |

1902. Ueber neue und bekannte Trematoden aus Seeschildkroten.

Zool. Jahrb. Syst., V. 16, pp. 524-694.

LuUue, Max.

1909. Parasitische Plattwtirmer, in Brauer, Die Stisswasserfauna

Deutschlands. V. 17, p. 24.

SKRJABIN, K. I.

1913. Tracheophilus sisowi n. g., n. sp. Ctrbl, Bakt., Abt. V. 69, pp.

90-95.

A MONOSTOME LUNG FLUKE

EXPLANATION OF PLATES

All drawings were made with the aid of a camera lucida

Cirrus

Excretory canal

Excretory pore

Excretory reservoir

Ganglion

Genital pore

Intestine

Nerve

Oral sucker

Ovary

Pharynx

ABBREVIATIONS

Prostate gland

Shell gland

Sperm mass

Seminal vesicle

Testis

Uterus

Vitellarium

Vagina

Vitelline duct

Vas deferens

Fig.

Oe pe et ose

a ae

FRANKLIN D. BARKER AND SUSANNE PARSONS

PLATE VI

Anterior end, ventral aspect; mass of sperm issuing from genital

pore.

Aorchis extensus Barker and Parsons, ventral aspect of small in-

dividual.

Excretory system from reconstruction.

Details of genitals from toto mount, dorsal aspect.

Egg.

Aorchis extensus Barker and Parsons, ventral aspect of large in-

dividual.

Pirate VII

Sagittal section of anterior end, showing relation of female genitals.

Cross section of body thru ovary, anterior to the testis.

Frontal section of anterior end showing self fertilization.

Sagittal section showing relation of male genitals.

Cross section of body thru pharynx.

Cross section of body thru testis and shell gland, posterior to the

ovary.

FIB+¢SAP deb 2

PRAtTE VI

ON ores

TOK yO.

“snes «

OVO

FIB+S. RR del 2

PLATE VII

“a

~~. *

We wd he

= Aas + ; v ee

Pe : > é, Pa ;

fe Wo. ' 2

ae ahd ee Son iiss

Fein ea ey i

eS waa as

7 rg 7 > \ An

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ars

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a ro ply

Pos

ENCHYTRAEIDAE (OLIGOCHZETA) FROM THE ROCKY

MOUNTAIN REGION*

By Paut S. WELCH

For certain groups of animal life, North America is still in

reality an unexplored territory. Of the vast region extending

from the Mississippi River to the Pacific and from the Isthmus to

the Arctic Ocean, excluding a narrow, coastal strip from Mexico to

Alaska, nothing has hitherto been known concerning the Enchy-

treide although there is every reason to believe that their distribu-

tion includes the entire area. The tendency of this family of

Oligochzta to be northern in its distribution is well established and

some of its representatives are known to thrive under alpine condi-

tions, both in Europe and North America. The two species dis-

cussed in this paper were found in high altitudes of the Rocky Moun-

tain system—mere chance representatives, no doubt, of a large in-

digenous enchytreid fauna which awaits study.

Mesenchytreus altus n. sp.

The following study has been made on material collected by

Professor Frank Smith near Corona, Colorado, close to the edge

of a small lake on the east side of Mt. Epworth. This lake, known

locally as ‘‘Pumphouse Lake,” is less than a mile from the point

where the Moffat railroad passes over the crest of the “Divide,” via

Rollins Pass, and has an altitude of about 11,300-11,400 feet. The

specimens, secured July 18, 1916, at and under the edge of a big

snow-field close to the north end of this lake, contained twenty-

nine sexually mature enchytrzids and nineteen others in various

stages of immaturity. All of the specimens were collected in moss

and under old wood. They occurred in such abundance that the

possible rate of collection was estimated at 50-100 per hour. Under

the conditions of their habitat, they were sluggish, but responded to

*Contribution from the Entomological Laboratory, Kansas State Agricultural Col-

lege, No. 28.

68 PAUL S. WELCH

contact stimuli and crawled slowly when disturbed. Although none

of them was found on or in the snow, they occurred in abundance

under it. While these worms cannot strictly be included with the

“slacier worms” described from certain other parts of the Rocky

Mountain region, yet it appears that their habitat represents, in

part, a combination of the conditions of the snow-field and the ter-

restrial environments.

Definition.—Length, 13-21 mm.; average, 17.5 mm. Diameter,

about 0.61 mm. Somites, 58-73; average, 63.7. Color of alcoholic

specimens, yellowish to light brown. Prostomium blunt, smooth,

rounded. Head pore at tip of prostomium. Sete sigmoid; ap-

proximately uniform in size and shape; 5-8 in ventral bundles in

anteclitellar region, 3-5 in lateral bundles; 2-5 in ventral bundles

in postclitellar region, 2-3 in lateral bundles. Clitellum distinctly

developed, on XII-XIII. Brain about twice as long as wide; lateral

margins almost parallel; anterior and posterior margins distinctly

emarginate. Origin of dorsal blood-vessel in XVIII; small cardiac

body present. Nephridia of usual mesenchytreid type; small ante-

septal, and large, irregular postseptal part; origin of efferent duct

on ventro-caudal surface of latter. Spermiducal funnel large, con-

torted; about five times as long as diameter; collar absent. Sperm

duct very long, extending caudad within ovisac to XIX; much con-

torted throughout entire length; diameter approximately uniform.

One pair of ovisacs extending from XII/XIII to XXII-XXVI;

both independent from origin; contain sperm sacs and sperm ducts.

One pair of sperm sacs extending caudad from XI/XII to XV-

XVII; vary somewhat in length; each contained within correspond-

ing ovisac. Penial bulb large, subglobular; atrium small, eight atrial

glands present; numerous groups of multicellular glands within

bulb. Spermathece elongate, extending to VII-VIII; ducts narrow,

no glands at ectal openings; ampulle long, terminations expanded ;

no connection with digestive tract.

The type and eight paratypes are in the collection of the writer.

Paratypes have also been deposited in the collection of the United

States National Museum and in the collection of Professor Frank

Smith.

ENCHYTR&IDZ (OLIGOCH ATA ) 69

Affinities —So long as a considerable number of foreign species

remain so briefly described, attempts to indicate relationships must

of necessity be attended by uncertainty. It appears that Mes. altus

differs from all of the foreign forms, as they are described at

present, in the possession of the enlarged and elongated spermathece,

such organs being apparently confined to North American species.

The structure of the spermathece puts the species into the group

composed of asiaticus, harrimani, setchelli, franciscanus, obscurus,

maculatus, vege, orce, and gelidus, differing from these mainly by

the complete absence of diverticula. The character of the ovisacs

and sperm sacs allies it with harrimani, fuscus, fuscus var. mermis,

gelidus, and possibly certain others not described in sufficient detail

to permit definite conclusions. With reference to the structure of

the penial bulb, it resembles, to varying extents, grandis, vege,

setchelli, maculatus, obscurus, and eastwoodi. It does not seem

possible at present to specify with any further accuracy the affinities

of this species. It should be noted that all of the above-mentioned

species are known only from the Pacific coastal region of North

America.

External Morphology

The body of Mes. altus is elongate, smooth, slender, cylindrical,

and tapers slightly in the region of the two extremities. The length

in the mature, alcoholic specimens varies from 13 to 21 mm., the

average being 17.5 mm. In the region of the clitellum, the diameter

is about 0.61 mm., the variation depending, at least in part, on the

degree of body contraction. The intersegmental grooves are more

distinct anterior to the clitellum and in the immediate region of the

caudal extremity, but are obscure throughout the remainder of the

body. The number of somites varies from 58 to 73, the average

being 63.7. Certain mature specimens have only about 40, but an

examination of the posterior end showed indications of loss of the

terminal somites and such individuals were not regarded as repre-

sentative. No pigment occurs in the body-wall or in the internal

organs, the body as a whole being sub-opaque, with a slight yellow-

ish or brownish color. The clitellum occurs on XII-XIITI and is

continuous on all aspects of these somites. It is well developed,

70 PAUL S. WELCH

its limits are distinct in the mature specimens, and the component

cells are approximately uniform in length, except in the region

about the penial bulb invaginations where they are somewhat shorter.

The prostomium is blunt, smooth, rounded, and bears a small, incon-

spicuous head pore on its apex. The sete are distinctly sigmoid

and disposed in the usual way in this genus, They are so deeply

set in the body-wall that, in some cases, it was found impossible to

determine accurately the number per bundle from whole, cleared

specimens, owing to the fact that sete are sometimes present but

almost entirely within the body-wall. It was necessary to depend

upon serial, transverse sections for accurate counts. In the ventral

rows, the number of sete per bundle varies from 5 to 8, usually 6

or 7, anterior to the clitellum, and from 2 to 5 posterior to the

clitellum, usually 4 or 5. In the lateral rows, anterior to the clitel-

lum, there are 3-5 sete per bundle, and 2-3 per bundle behind the

clitellum. Setz are present on XIII but are not specialized. In

mature specimens, the position of each penial bulb invagination is

made prominent by the protruding margins.

Internal Morphology

Brain.—The brain lies in I and II, principally in the former.

The length is about twice the width, a typical measurement being

as follows: length, 0.126 mm.; width, 0.054. Both anterior and

posterior margins are distinctly emarginate and the lateral margins

are approximately parallel. Two pairs of supporting strands ex-

tend from the lateral margins to the body-wall.

Blood Vascular System.—The dorsal blood-vessel arises in

XVIII and is distinctly swollen in each of the somites posterior to

the clitellum. A small cardiac body is present.

Nephridia—The nephridia are of the typical mesenchytraid

type. The anteseptal part is merely a nephrostome, while the post-

septal part is large, irregular, somewhat lobulate, with the efferent

duct arising from the ventro-caudal surface. The organ as a

whole shows a considerable amount of variation in size, shape, and

general proportions.

Spermiducal Funnel—The spermiducal funnels are conspicu-

ous organs in the clitellar region. Each is long, cylindrical, con-

ENCHYTREIDE (OLIGOCH&TA) 71

torted, of approximately uniform diameter and without a collar at

the free extremity. Each funnel is about five times longer than

wide and, in some of the specimens examined, extends through both

XI and XII. Not only do they vary in degree of contortion but the

position in the body cavity is not constant. There is a general

tendency for one or both to be located caudad of the usual position.

In certain specimens examined, one funnel is in the usual position

while the corresponding one on the opposite side is farther caudad

in XII and XIII. Specimens were studied in which both funnels

are in XII and XIII. Sections showed that sometimes a funnel

is shifted from its usual position in XI back into the anterior part

of the sperm sac.

The sperm duct is very long, extending caudad as far as XIX.

It is very much contorted throughout the entire course so that its

true length is not represented by the number of somites through

which it extends. Each lies in the ovisac of the same side of the

body, but not within the sperm sac.

Sperm Sacs and Ovisacs.—lIn this species is found a pair each

of sperm sacs and ovisacs which are intimately related in their

mode of origin and general position. The sperm sacs are formed

by caudal reflections, a right and a left, of the septum XI/XII to

form two hollow tubes into which the maturing spermatozoa pass.

These sacs vary in the different specimens with respect to the length

and amount of distension. The two sacs of the same individual may

not be equal in length. In the specimens studied, the sperm sacs

extend caudad to XVI-XVII. Each sac is contained within the

corresponding ovisac.

The ovisacs are extensive, hollow tubes which arise as caudal

reflections, a right and a left, of the septum XII/XIII. Both are

independent from the beginning and each occupies a considerable

portion of the ccelom, laterad and ventrad of the digestive tract.

One may be longer than the other in the same specimen but both

extend well caudad. In the specimens examined, the posterior

extent was found to be XXII-XXVI. Each ovisac contains the

sperm duct and the sperm sac for the same side of the body, and,

in addition, contains the developing egg masses.

72 PAUL S. WELCH

Penial Bulb.—A pair of large penial bulbs occurs in the usual

position in XII. In structure, they conform to Eisen’s mesenchy-

treid type (’05, p. 7) and consist essentially of two sets of struc-

tures, the penial bulb proper and the atrium with its associated parts.

In the fully retracted condition, the two bulbs, including the penial

invaginations, occupy more than one-half of the coelom in XII.

The penial bulb is situated on a deep invagination, lined by a

continuation of the external hypodermis and cuticula. In transverse

section of the body, this invagination is slit-like, except at the mesal

extremity where it expands into a foot-shaped enlargement. The

transition from the external hypodermis to the lining of the invagi-

nation is accompanied by less reduction in thickness than is the usual

condition in enchytreids. The internal structure of the bulk proper

has the characteristic complexity of the mesenchytreid type. Many

lobular, multi-cellular penial glands occupy most of the interior,

each opening into the penial invagination. They are so closely

massed together that an accurate count is almost impossible. All

have essentially the same structure, although considerable variation

occurs in size and shape. Accessory glands appear to be absent.

Numerous muscle strands radiate through the interior of the bulb,

extending among the penial glands. While these glands occur

throughout the organ, the majority of them lie in the dorsal and

mesal parts. The usual retractor muscle extends latero-dorsad from

the innermost tip of the penial invagination to the body-wall. A

thin, delicate peritoneum forms the envelope for the whole organ.

The atrium unites with the penial bulb at the dorso-mesal part,

opening into the penial invagination. It is only moderately devel-

oped, rather short, spindle shaped, and merges gradually with the

sperm duct a short distance from the margins of the penial bulb.

The greater part of the atrium is within the latter. From the ectal

end, which protrudes into the penial invagination, it extends meso-

dorsad through the bulb and beyond it for a very short distance,

then bends caudad to meet the sperm duct. The total length is not

more than that of the fully retracted penial invagination, and the

maximum diameter is only about 0.08 mm. It is thick walled, the

thickness being due largely to the well-developed longitudinal muscle-

layer which extends the full length of the atrium. The maximum

ENCHYTREIDE (OLIGOCHATA ) 73

thickness of this muscle-layer is about 0.016-0.02 mm. A narrow

zone of deeply staining nuclei occurs just entad of this muscle-layer,

the exact nature of which has not been determined. The lining of

the lumen of the atrium is composed of a single layer of large, almost

clear cells, nucleated at the bases and ciliated on the exposed sur-

faces. Well-developed atrial glands are present and, in the speci-

mens examined, the number is eight. They all unite at the same

level with the upper part of the atrium, each entering independently.

The point of union is very close to the margin of the penial bulb,

in fact, the glands and their ducts are so closely associated with the

bulb that a critical examination of sections was required in order to

distinguish them. ‘They are large, multicellular, pyriform organs,

somewhat irregular in shape and varying slightly in size. They do

not extend out loosely into the ccelom but are closely applied to

the penial bulb. The ducts are very small and easily overlooked, the

best view being found in sections made at right angles to the lumen

of the atrium. At first sight, they appear to be large penial glands,

but more careful examination shows that the staining reaction of the

two groups of glands is not the same.

Spermathece.—This species belongs to the group having long,

prominent spermathece which extend through a number of somites.

The ectal opening occurs in the usual position in IV/V and is char-

acterized by a conical thickening of the cuticula above the aperture

and the absence of associated glands. Unlike many enchytreids,

the spermathece of this species occur entirely ventrad of the alimen-

tary canal, and in all of the specimens examined, they cross over

about midway of their length so that in the somites containing the

terminations of these organs the ampulla on the right side really

belongs to the left spermatheca, and the ampulla on the left to the

right spermatheca. Diverticula are absent and it is difficult to deter-

mine the precise extent of the duct and the ampulla. However, each

organ is usually present in V as a narrow duct, expanding in VI or

VII to form the ampulla. Both organs end blindly and have no

connection with the digestive tract. The terminations are distinctly

distended and in the mature individuals contain a considerable quan-

tity of spermatozoa. The two organs of the same individual vary

somewhat in extent, usually one being shorter than the other. In

74 PAUL S. WELCH

some instances, the spermatheca on one side extends into the poster-

ior part of VIII while the opposite one ends in VII. None of the

spermathece examined extend beyond VIII.

The writer pointed out in a previous publication (’16, pp.

99-100) that as judged from the literature, enlarged and greatly

elongated spermathece, similar to the ones described above, have

been found only in certain American species from the Pacific Coast

region. At present, the list includes seven species described by

Eisen (05): Mes. harrimam, Mes. setchelli, Mes. franciscanus,

Mes. obscurus, Mes. maculatus, Mes. vege, and Mes. orce. Pos-

sibly Mes. asiaticus Eisen should also be included. To this list

should be added Mes. gelidus Welch and Mes. altus, n. sp. It ap-

pears that the spermathece of the latter differ from those possessed

by any of the above-mentioned species in the total absence of diver-

ticula. Aside from Mes. orce and Mes. asiaticus which have dimin-

utive spermathecal diverticula, the other species have diverticula as

well-developed and prominent organs. Further discussion of en-

larged spermathecz occurs in the writer’s previous paper (’16, p.

100).

An interesting feature of the spermathecz of this species is

the crossing over described above. All of the specimens examined

showed the same condition, indicating that it is apparently the nor-

mal form for the species. Since these organs end blindly and are

not connected with the digestive tract, some variation in the posi-

tion of the terminations with reference to the other internal organs

might be expected. However, the fact that they extend through

several somites separated by the usual septa indicates that probably

this crossing over occurred when the organs were developing, rather

than after their full length had been attained. This interpretation

is supported by the fact that the crossing over seems to be constant

in character. Additional support also appears in the discovery by

Eisen (’05, pp. 102-104) that in some of the specimens of Henlea

guatemale the spermathece cross over before effecting their inde-

pendent union with the digestive tract, a condition which could have

arisen only in connection with the development of the organs.

ENCHYTREIDEZ (OLIGOCH ETA) 75

Henlea urbanensis Welch

This species was originally described by the writer (714, pp.

134-140) from a single, sexually mature specimen, found near Ur-

bana, Illinois, in the rich soil of undisturbed forest-land, and, up

to the present date, nothing further was known as to its distribu-

tion. Through the courtesy of Miss Bessie R. Green, the writer

has recently been able to study a collection of enchytrzids which

were collected in a limited locality on the shore of Yellow Bay,

Flathead Lake, Montana, on July 20, 1914. These worms occurred

in the moist earth under moss on the banks of a small stream located

near the University of Montana Biological Station. They were

also found on logs which had fallen into the stream. The sur-

rounding region is wooded and has an elevation of about 3,000 feet.

The stream is a typical mountain one, being clear, cold, and rather

swift. This collection contained five completely mature specimens

and five others in various states of immaturity. Critical examina-

tion of serial sections made from mature individuals revealed so

close a correspondence with the description of the type specimen

of Henlea urbanensis that they are without question the same species.

Any of the deviations from the original description can easily be

accounted for as variations within the same species. The following

discussion of the chief morphological features will indicate these

variations and extend the description as originally given.

External Morphology

The length of the mature specimens varies from 10 to 15 mm.,

the average being 12.5 mm. It will be noted that this dimension

falls considerably below that of the type specimen, but a part of

this discrepancy may be due to contraction of the animals during the

killing and fixing process. It is also possible that the Montana speci-

mens are somewhat smaller. In any case, this particular difference

cannot be regarded as specially significant. The maximum di-

ameter, which occurs in the clitellar region, is about 0.61 mm. The

number of somites varies from 50 to 53, the average being 51.8.

Owing to the lack of evidence as to the number of somites of the

type specimen, no comparison of these data can be made. The color

76 PAUL S. WELCH

of the alcoholic material is light yellowish to light brown. The

prostomium is rounded on the tip, somewhat blunt, short, concave

on the dorsal surface, and bears a distinct head pore at 0/1.

On the anterior and middle parts of the body, there are 4-8 sete

per bundle, 5-7 being more common, while on the posterior region,

the number decreases to 2-4.. All of the sete of the body are ap-

proximately straight, both at the distal and proximal extremities.

In connection with the reexamination of the type specimen from

Urbana, it was discovered that the statement in the original descrip-

tion (714, p. 135) that the proximal ends of the sete are “distinctly

bent” requires some modification since they are approximately

straight in form, the outer sete only of each bundle deviating from

the strictly straight condition by bending very slightly away from

the central axis of the bundle. The outer sete of each bundle are

longer and stouter than the inner ones, a contrast which recalls the

condition in the genus Fridericia, although the difference is less than

that usually found in the latter.

A well-developed clitellum occurs on XII-XIII and sometimes

to some extent on the posterior part of XI. It is continuous around |

the body, showing practically no diminution of thickness on the

ventral surface. The component cells are closely crowded together

and in the thickest region are 5-6 times longer than thick. The

clitellum on the type specimen corresponds exactly with those on

the Montana specimens with the exception that its thickness is some-

what greater.

Internal Morphology

Lymphocytes—The lymphocytes are large, broadly oval in

shape, very numerous, and scattered throughout. the whole body-

cavity. Each is distinctly nucleated and is composed of a relatively

large amount of granular cytoplasm. Although there is some varia-

tion in size and shape, there appears to be only one type. The maxi-

mum diameter varies from about 0.04 to 0.056 mm.

The above-mentioned characters apply equally well to the

lymphocytes of the type specimen from Urbana. In a few somites

of the latter, cephalad of the clitellum, they are so numerous that

they almost completely fill the ccelom.

ENCHYTREIDE (OLIGOCH ATA) 77

Brain—Every detail of the original description holds for the

Montana specimens and no additional comment is required.

Peptonephridia—In general structure, extent, and relation to

the digestive tract, these organs do not differ to any degree from

those of the type specimen. There is a slight variation in the num-

ber and size of the branches, which, however, cannot be regarded

as very significant. They arise from the alimentary canal in V and

extend caudad to VIII. Both dorsal and ventral organs are com-

paratively simple, apparently single and tubular. Sparse branching

occurs at one or two points along their course.

Intestinal Diverticula—Critical examinations of serial sections

made in transverse and frontal planes show that the structure of

the intestinal diverticula of the Montana material corresponds almost

exactly with that of the same organs in the Urbana specimen. Since

the latter was described in detail (Welch, °14, pp. 137-139), no

morphological discussion will be given here. The paper just men-

tioned contains a structural comparison of some of the various intes-

tinal diverticula which occur in species of Henlea and it is pointed

out that, so far as could be judged from the literature, the mor-

phology of the intestinal diverticula of H. urbanensis differs from

that of any species then known. However, since that time, Friend

(715, pp. 203-204) has described, in some detail, the structure of

the pair of diverticula occurring in H. fragilis which corresponds

very closely to those of H. urbanensis, the only important differences

being the absence of chloragog cells and the presence of a larger

number of internal folds on the mesal side. A pair of diverticula

are said to occur (Southern, ’09, p. 146; Friend, 12, pp. 577-598)

in VII or VIII in H. hibernica Southern, H. attenuata Friend, H.

heterotropa Friend, H. fridericioides Friend, and H. triloba Friend,

but the structure has not been described for any of them, making

comparison impossible.

Dorsal Blood-vessel—The dorsal blood-vessel arises in the

extreme posterior part of VIII or the anterior part of IX and is

distinctly enlarged at the origin. It is evident that there is not an

exact correspondence with the position of origin as given for the

type specimen but such slight variation is, no doubt, within the limits

of a species.

78 PAUL S. WELCH

Penial Bulb.—The structural detail of this organ corresponds

so closely with the original description that only one or two varia-

tions deserve comment. In the Montana specimens, the cells com-

posing the body of the bulb are not so distinctly differentiated into

the three kinds previously described, although they appear to be

represented. The series of cells arranged radially about the penial

lumen is not as complete and the gland cells composing the dorsal

part of the bulb differ but slightly from those of the ventral part.

Spermathece.—The spermathecz lie in the usual position in V,

the ectal opening of each being laterad in the intersegmental groove

IV/V and the ental opening at the posterior part of V on the dorsal

side of the digestive tract. Each opens independently into the lat-

ter. A distinct rosette of glands occurs about the ectal opening.

Viewed as a whole, the organ shows no differentiation between the

duct and the ampulla, having a maximum diameter of about 0.148

mm. near the ectal opening and gradually decreasing in size to the

union with the digestive tract. An examination of sections of this

organ shows that about midway of its length there is a distinct ex-

pansion of the lumen of the organ, probably representing the ampulla

although its presence is not indicated externally. In other parts of

the organ, the lumen is very fine. The presence of this expansion

of the lumen indicates the extent of the duct and a typical set of

measurements is as follows: length of duct, 0.435 mm.; length of

expanded portion of lumen, 0.348 mm.; length of remainder of or-

gan, 0.957 mm. ; total length of ampulla, 1.305 mm.

It will be noted that the only difference between the sperma-

thecee of the Montana and the Urbana material is the absence in

the former of the slightly expanded union of the two organs just in

advance of their fusion with the digestive tract. While all of the

essential parts of a spermatheca are represented in these organs,

nevertheless they appear to have a very limited capacity and are

rather inconspicuous features of the internal morphology. At first

sight, it might appear that these spermathece are not completely

developed, but in all of the sexually mature specimens examined,

they were constant in shape and degree of development. Further-

more, they were found to contain masses of spermatozoa, indicating

that they were at least functioning in the usual capacity of com-

ENCHYTREIDE (OLIGOCHATA ) 79

pletely mature spermathece. Similar spermathece have been re-

ported in H. californica Eisen and its two varieties, monticola and

helene, and in H. tenella (Eisen). Certain other briefly described

species appear to have similar spermathece.

Nephridia and Spermiducal Funnel.—These organs correspond

to the previous description and require no comment here.

Affinities —A critical examination of the literature shows that

of the various species usually included under the genus Henlea, the

following apparently approach H. urbanensis rather closely: nasuta

(Eisen), tenella (Eisen), californica Eisen, inusitata Friend, fragilis

Friend, and fridericioides Friend.

Nasuta fails to agree with urbanensis by possessing spermathece

with distinct ampulle and in having the structure of the ectal and

ental walls of the intestinal diverticula quite similar. The writer

has found nothing in urbanensis to correspond to the statement by

Southern (’09, p. 146) concerning an Irish specimen of nasuta:

“The single specimen obtained was very dark, each segment having

several rows of irregular glands.”

Tenella; an imperfectly described species from the Old World,

seems to agree closely with urbanensis in all of the structures de-

scribed except the small number of sete and the possession of a

spindle-shaped spermiducal funnel. A number of important struc-

tures are not mentioned.

Californica differs from urbanensis in having a narrow and

pointed prostomium, no chloragog cells on the digestive tract or on

the dorsal blood-vessel, the width of the brain exceeding the length,

the presence of glands at the ectal opening of the spermathece,

and the position of the intestinal diverticula in VII. Two varieties,

monticola and helene, have been described for californica by Eisen

but the above-mentioned differences hold for them also, except

that the brain in helene is described as being almost square.

Inusitata (Friend, 713, pp. 83-84; ’14, p. 136) has a single in-

testinal diverticulum (“bulb”) and the salivary glands are “neph-

ridia-like glands” between the second and third septal glands.

Fragilis seems to be very close to urbanensis, differing in the

character of the spermathece, the former having well-developed

ampullz and each member of a pair is independent of the corre-

80 PAUL S. WELCH

sponding one on the opposite side. The union with the digestive

tract is described as incomplete, the ental end of the ampulla end-

ing blindly in the wall-tissues of the tract. Fragilis also shows the

structure of the ental and ectal walls of the intestinal diverticulum

to be similar, and no chloragog cells occur on the ccelomic surface

of the latter. Chloragog cells are also absent from the first twenty

to twenty-five somites.

Fridericioides also seems to bear considerable resemblance to

the species under discussion. Although not completely described,

the known details of internal and external anatomy indicate a differ-

ence in the presence of a well-defined ampulla on each spermatheca.

Each spermatheca opens independently into the digestive tract and

has a series of glands at the ectal opening. Friend (’12, p. 587)

states that the septal glands sometimes consist of two pairs only.

Three pairs are present in urbanensis in the usual position and all

are well developed. However, it is not known how much weight

can be attached to variation in these glands. The structure of the

pair of intestinal diverticula is not described.

It will be noticed that some of the above-mentioned species,

as judged from descriptions and figures, seem to resemble urbanensis

rather closely. It is very possible that not all of these species are

valid and the writer is inclined to expect that future revision based

on more complete morphological data will place some of them in

the synonymy. Some of the apparent differences or similarities

may be the result of incomplete knowledge of variation within each

species or of certain structural features which are important in diag-

nosis. The close agreement of the Montana specimens with the

original type specimen from Illinois is of interest because of the

wide separation of the two regions.

ENCHYTREIDA (OLIGOCH ETA) 81

LITERATURE CITED

EIsEN, G.

1905. Enchytreide of the West Coast of North America. Harriman

Alaska Expedition, 12:1-166. 20 pl. New York.

FRIEND, H.

1912. British Enchytreids. IV. The Genus Henlea. Journ. Royal Micr.

Soc., pp. 577-598. 11 figs.

1913. A Key to British Henleas. The Zoologist (4), 17:81-91.

1914. British Enchytreids. VI. New Species and Revised List.

Journ. Royal Micr. Soc., pp. 128-154.

1915. Studies in Enchytreid Worms. Henla fragilis Friend.

Ann. Applied Biol., 2:195-208. 6 pl.

SOUTHERN, R.

1909. Contribution towards a Monograph of the British and Irish

Oligocheta.

Proc. Royal Irish Acad., 27, Sec. B:119-182. 5 pl.

WELcH, P.S.

1914. Studies on the Enchytreide of North America.

Bull. Ill. State Lab. Nat. Hist., 10:123-212. 5 pl.

1916. Snow-field and Glacier Oligocheta from Mt. Rainier, Washington.

Trans. Am. Micr. Soc., 35:85-124. 4 pl.

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THE MORPHOLOGY, STRUCTURE, AND DEVELOP-

MENT OF HYDRACTINIA POLYCLINA

By J. A. PLAce

With Plates VIII and IX

CoNTENTS

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On CGP ACE NCEE). So ie OY SQN Jah my kh) oY CaaS 86

SOR EAA OPOUIGS | ccs een. Oe Ue ou hea Sora 86

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PANEL V OV Perea wy re oe es wie A eeee he Cal eA re oss ws Peal iphigee 88

MOOMRSMNI Sas tetltle a PU LEY ag Letom cake te etna Evie cc el da rasan ELSIE 89

PD OUT ane tat oe Sean hl seca ie Ati Y cat une RSI hang RK 90

INTRODUCTION

During the summer of 1914, the Invertebrate Zodlogy class

at The Marine Biological Laboratory, Woods Hole, while working

on fHydractinia polyclina, experienced some difficulty in finding,

and identifying Tentacular polyps. The question arose as to their

constancy in the colony, and it was suggested by the instructors

as a good subject for special work. The purpose of this paper

is to offer a brief account of the morphology, structure, and de-

velopment of the various kinds of polyps that exist in a colony of

Hydractinia polyclina.

This work was carried on in the Department of Biology at

Ohio University. I wish to thank Dr. W. F. Mercer, head of the

department, for many valuable suggestions and for his generosity

in giving us free access to the laboratory equipment. I also wish

to thank Mr. J. T. Pickering, assistant in the department, for his

assistance in preparing the accompanying figures.

84 J. A. PLACE

HABITAT

Hydractima polyclina lives in a state of symbiosis with the

Hermit-crab, forming over the shell of the latter a soft, pinkish

covering. It appears that both are benefitted by this community

life, for while the Hydractinia colony is furnished with transpor-

tation it, in turn, affords protection to the Hermit-crab, not only

by obscuring its shell from view, but also by its possession of sting-

ing cells by means of which it forms a defense against the enemies

of the Hermit-crab.

The constant association of these forms together led to the

belief that the life of the Hermit-crab was necessary to the exist-

ence of Hydractinia. That Louis Agassiz found them growing

in abundance, attached to rocks in tide pools; that Samuel F. Clarke

later found them growing on a wharf at Fort Wool, Chesapeake

Bay; that two students of Woods Hole Laboratory found them

growing on Mytilus edulis and Limulus; that during the summer

of 1891, Dr. Conklin found them growing on the Fish Commission

Wharf, Woods Hole; that they are frequently found growing on

sponges about Woods Hole region; have proved their existence in-

dependently of the Hermit-crab. Besides having removed the

occupants from shells bearing Hydractinia colonies, we placed the

shells in wire baskets, and suspended them below low tide from a

wharf. There we left them three weeks during which time the

colonies became very luxuriant on the empty shells.

An additional advantage received by the Hydractinia colonies

is that of food supply furnished by the young paguri, Miss Bunt-

ing reports that many of these are devoured by the polyps as they

swim out from the maternal shell.

MATERIAL AND METHODS

The material for this work was obtained while studying at the

Marine Biological Laboratory, Woods Hole, during the summer

of 1914. The shells possessing Hydractinia colonies were found

at low tide in Eel Pond. In order to prevent the polyps from

contracting into abnormal shapes they were narcotized by adding,

drop by drop, a solution of ten per cent chloreton in absolute al-

HYDRACTINIA POLYCLINA 85

cohol till all power of contraction was lost. They were preserved

in a four per cent formalin solution. For histological structure

the best results were obtained with iron-hematoxylin stain. Sec-

tions were made from seven to twelve micra in thickness. Camera

lucida drawings were made of various magnifications.

THE POLYPS

The gasterozooids, as a rule, are the most numerous repre-

sentatives of the colony; but, sometimes, during the summer months

reproduction is so active that the blastostyles, occasionally, are

equally as numerous. These are the longest of the polyps often

exceeding one-fourth inch in length. They possess a conical hypo-

stome terminating in a large mouth. Around the base of the hypo-

stome are two circles of tentacles which increase in number with

age from ten to thirty. The longest tentacles that occur on mem-

bers of the colony are found here; they are crowded with nemato-

cysts. The external surface is covered with a layer of ectoderm

which is continuous with the upper ectoderm of the ccenosarc.

Since it is the function of this polyp to collect food for the

entire commonwealth, it possesses the largest gastral cavity (PI.

VIII, Fig. 3). It is lined with a single layer of endoderm continuous

with that of the endodermal canals of the ccenosarc.

The ectoderm and endoderm are separated by a thin structure-

less layer of mesogloea which will not be referred to in description

of the other polyps as it is common to all.

The blastostyles are either male, or female, though both sexes

are never found in the same colony. The mouth, and gastral

cavity are both small. A short distance below the mouth are two

circles of tentacles varying in number from ten to thirty, but, un-

like those of the gasterozodids, are very rudimentary consisting

of knob-like structures crowded with nematocysts. Immediately

below the head the walls constrict into a narrow neck and, then,

enlarge into a globular dilatation from which arise the sporosacs. In

both sexes the reproductive cells arise in the body and migrate to

the sporosacs. In female colonies the sporosacs are filled with

eggs that can be seen through the thin ectodermal walls in the un-

stained, as well as stained, condition (Pl. VIII, Fig.2). Inthe male

86 J. A. PLACE

colonies the reproductive elements are small, very numerous, and

stain more deeply than the body cells (Pl. VIII, Fig. 4). Below the

sporosacs the body again narrows often into a slender thread.

The difference in appearance between the male and female

colonies—the male sporosacs being often much elongated and of

a yellowish tint while the female are rounded and rose-colored—

caused Van Beneden to regard them as two distinct species which

he described in a paper published in 1844 as Hydractinia lactea and

Hydractinia rosea.

The dactylozodids are about the same size throughout their

length. Their distal extremity is surrounded by a circle of rudi-

mentary tentacles from ten to sixteen in number. There is a very

small mouth in the center. These have strong, muscular walls and

are capable of coiling and uncoiling themselves. As their function

is chiefly to protect the other members of the colony they possess

an abundance of nematocysts (Pl. VIII, Fig. 1).

The tentaculozooids are extremely slender though often exceed-

ing the dactylozooids in length (Pl. VIII, Fig.5). They are capable

of great extension and are characterized by Mr. Hincks as floating

like long fishing lines through the water. In preserved material,

on the other hand, they are contracted to such an extent as to ren-

der them extremely difficult to find. They are situated near the

outskirts of the colony, and are usually few in number as com-

pared with other members. The tip, only, is covered with nemato-

cysts. No mouth is present, and the gastral cavity is very small.

These were regarded by Allman as abnormal dactylozodids on

account of their paucity. Colcutt, however, found them present

in every colony of Hydractinia echinata and considered them as

normally present; Mr. Hincks reports them as constantly occur-

ring. He also states that he distinguishes no difference between

his Hydractima echinata, and Hydractinia polyclina of Agassiz.

I have found the Hydractinia polyclina of Woods Hole region to

correspond in every particular with Colcutt’s Hydractinia echinata,

but the skeleton differs in minor details from that of Hydractinia

echinata as described by Mr. Hincks to which reference is made

under the discussion of the skeleton.

HYDRACTINIA POLYCLINA 87

THE SKELETON

The skeleton is a chitinous structure which forms an irregular

crust on univalve shells, or other objects on which the colony is

growing. The skeletal structures penetrate the shell by dissolving

the calcareous substances with an acid, or erosive agent which the

animal secretes. The chitin is then secreted by the lower ectoderm

of the ccenosarc in thin layers (Pl. IX, Fig. 4). These are so closely

attached to the shell that the latter must be dissolved away with

dilute hydrochloric acid in order to obtain good specimens of skele-

ton. Pieces of skeleton can then be cut from the shell and thin

sections made.

The skeleton is overlaid by ccenosare consisting of two layers

of ectoderm enclosing between them a number of endodermal tubes

which branch, and anastomose promiscuously. These are connect-

ed at intervals with the canals of the polyps whose ectoderm, and

endoderm are continuous respectively with the upper ectoderm of

the ccenosarc, and the endoderm of the tubes. In this way the gas-

tral cavities of all members of the colony are placed in direct com-

munication with each other.

At intervals, the skeleton projects above the ccenosare forming

conical smooth spines, and spinules. These sometimes form bridges

of chitin over an intercommunicating tube which led Mr. Hincks

to conclude that the chitinous covering existed above, as well as be-

low the ccenosare (Pl. IX, Fig. 4).

Carter (1873) tells of a specimen in the British Museum in

which the whole of the shell has become transformed into the horn-

like skeleton of Hydractinia. From the smooth internal appear-

ance, he infers that the shell had been tenanted by an Eupagurus

which left after the entire shell had been transformed.

HISTOLOGY

The lower ectoderm of the ccenosarc is composed of long slen-

der cells quite irregular in shape. They are more or less vacuolated,

and contain a single nucleus situated near the center of the cell.

The nucleus is oval in shape, and contains several nucleoli. It is

88 J. A. PLACE

the function of this layer to secrete, extend, and renew the chitinous

skeleton.

The upper ectoderm of the ccenosarc is formed of a single

layer of cells more regular in size, and more cubical in shape.

Nematocysts are occasionally present in this layer. (PI. IX, Fig. 4) i

This layer is continuous with that of the polyps, the main dif-

ference in the latter being a greater variation in shape of their cells.

The endoderm of the ccenosarc is made up of a single layer of

cubical cells containing a single, oval nucleus in their center.

In the gasterozodids the endoderm contains long, narrow cells

which vary in length so that the free ends are not at the same level.

In this way longitudinal ridges are formed in the lumen which in

cross section present a very irregular appearance (PI. IX, Fig. 3).

As this is especially characteristic of the nutritive polyps, it is evi-

dent that even in this low form of life the rudimentary alimentary

canal is thrown into elevations for the increase of surface.

These cells are wider at their free ends, and are vacuolated.

Their nuclei are oval, are situated near the middle region of the

cell, and possess one or more nucleoli.

The endoderm of the blastostyles is composed of long, narrow

ciliated cells. In the head region these often contain several nuclei,

but in the body they possess a single, large nucleus.

The endodermal cells of the dactylozodids are approximately

equal in size. They usually contain many vacuoles, and a single

nucleus which is situated in the middle of the cell.

The tentaculozodids possess long, narrow cells which are more

regular in size and shape than those of the other polyps.

EMBRYOLOGY

The ova have their origin in any part of the endoderm below

the gonophores, and migrate upward between the ectoderm and en-

doderm until they reach the gonophores. Here they remain till

they ripen and are laid (Pl. VIII, Fig. 2).

The origin of the sperm cells is a little more complicated. At

the outset the ectoderm of the gonophore begins to divide into two

layers, the inner one of which stains deeply and is destined to form

the sexual cells. This layer, in turn, divides into two, an outer

HYDRACTINIA POLYCLINA 89

thin layer consisting of a single row of cells, and an inner one

which rapidly separates into several rows of cells. These stain

deeply and are known as spermatoblasts. They become specialized

to form the mature spermatozoa.

The ova are fertilized at the moment of ejection. The polar

bodies are rapidly given off, cleavage takes place, and a ciliated

planula is formed. This becomes attached at one end, elongates,

and tentacles are formed at the other. At the basal end prolonga-

tions are given off to form the beginnings of the tubular network.

_ These subdivide promiscuously, the intervening spaces being grad-

ually filled in by the extension of the ccenosarc, and the secretion

of spines and spinules.

CONCLUSIONS

1. Hydractinia polyclina, though almost invariably associated

with the Hermit-crab, is capable of an independent existence.

2. In every Hydractinia colony there are normally present

four kinds of polyps:

Gasterozooids.

Blastostyles.

Dactylozooids.

Tentaculozooids.

3. The function of the gasterozodids is to collect, digest, and

absorb food for the entire colony; that of the blastostyles is repro-

duction. The function of both the dactylozodids and tentaculo-

zooids is to defend the colony against enemies, the latter being

especially adapted to this service by reason of its great length.

4. Judging from the accounts of the many investigations made

upon Hydractinia echinata, we have concluded that it is identical

with Hydractinia polyclina.

Department of Biology,

Ohio University, Athens, Ohio.

90 J. A. PLACE

BIBLIOGRAPHY

CotcutTt, M. C.

On the structure of Hydractinia echinata. Journal Mic. Soc., Vol. 40,

p. 77-99, Pl. 1.

Brooks, W. K.

The life history of the Hydromeduse. Mem. Boston Soc. Nat. Hist. 3.

Agassiz, L. |

Contr. ‘Nat. Hist)’ 0. S.°4, p: 227.

BrecKwitH, Cora J.

The genesis of the plasma structure in the egg of Hydractinia echinata.

Journal of Morphology, June, 1914, p. 189-252.

BuntineG, M.

The origin of sex cells in Hydractinia and Podocoryne; and the develop-

ment of Hydractinia. Journal of Morphology, Vol. 9, p. 203-231.

Carter, H. J.

Transformation of an entire shell into chitinous atructtitie by the polype

Hydractinia, with short descriptions of the polypidoms of five other

species. Ann. Mag. Nat. Hist., 4th Ser., Vol. 11, p. 1-15.

Carter, H. J.

On new species of Hydractinide, recent and fossil, and on the identity

in structure of Millepora alcicornis with Stromatopora. Ann. Mag.

Nat. Hist., 5th. Ser., Vol. 1, p. 298-311.

Hincxs, THoMAS

A history of the British hydroid zodphytes, Vol. 1, Pl. 4, Vol. 2.

Goto, SEITARO

On two species of Hydractinia living in symbiosis with a Hermit- crab.

Journal Exp. Zoo., Vol. 9.

SMALLWoop, W. M.

A reexamination of the Cytology of Hydractinia and Pennaria. Biol.

Bull. 17, No. 3, p. 209-233. |

Fig.

a a te

weiled AG aor weet 2 Mine 5

HYDRACTINIA POLYCLINA

EXPLANATION OF PLATES

Pirate VIII

Dactylozodid x 96.

Female blastostyle x 96.

Gasterozodid a

Male blastostyle K-53;

Tentaculozoodid x 130.

A colony of Hydractinia polyclina growing on an Eupagurus shell.

PLATE IX

Cross section of male gonophore x 413.

Cross section of female gonophore x 413.

Cross section of gasterozodid, middle region x 413.

Cross section of skeleton xia

ABBREVIATIONS

Ch, chitinous skeleton. mes, Mesoglcea.

E, Egg. n, Nucleus.

En, Endoderm. n.c, Nucleolus.

ec, Ectoderm. R.T, Rudimentary tentacles.

G, Gonophore. S.c, Sperm cells.

G.C, Gastral cavity. Sp, Spine.

h, Hypostome. T, Tentacle.

L.Ec, Lower ectoderm. up.ec, Upper ectoderm.

L.En, Lower endoderm. up.en, Upper endoderm.

Mouth.

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ANOTHER CESTODE FROM THE YOUNG CAT!

J. E. Ackert and A. A. Grant

The ability of certain carnivore cestodes to develop in more

than one host suggested the question: Will the common dog tape-

worm, Tenia pisiformis Bloch, develop in the domestic cat (Felis

domestica).

Several kittens reared in cages covered with wire screen

(eighteen mesh) were given Cysticercus pisiformis from the peri-

toneal cavity of the cottontail, Sylvilagus floridanus mearnsu (in-

termediate host of this dog tapeworm).

In the experiment, three control and eight experimental kittens

were used. When kittens 22, 23, 24, 25, 27, 29, and 32 were ap-

proximately six weeks old, feedings of cysticerci were made every

few days until from ten (kittens 27, 29, 32) to approximately sev-

enty-five cysts had been fed to each kitten. The examinations of

these animals yielded the following results:

Kitten 27, examined two days after feeding, three tapeworms

(length, 3.5 and 4 mm.) in the small intestine.

Kitten 29 (died), examined two days after feeding, eight tape-

worms (length, 3.5 to 6 mm.) in the small intestine.

Kitten 32 (died), examined five days after feeding, five tape-

worms (length, 6 to 10 mm.) in the small intestine.

Kitten 24, examined twenty-one days after first feeding, two

tapeworms (length, 6 mm.) in the small intestine.

Kitten 25, examined twenty-two days after first feeding, three

tapeworms (length, 10 mm.) in the small intestine.

Kitten 23, examined fifty-two days after first feeding, four

tapeworms (length, 4, 5.5, 12.5, 22 mm.) in the small intestine.

Kitten 22, examined ninety-eight days after first feeding, no

tapeworms.

1Contribution No. 17 from the Zoological Laboratory, Kansas State Agricultural

College. Aid of Experiment Station.

94 J. E. ACKERT AND A. A. GRANT

Of the cestodes obtained from these kittens, two from kitten

23 were developed to the extent of having reproductive organs fully

formed. A description of them will be given later.

To ascertain in what part of the digestive tract evagination of

the cysticerci occurs, kitten 26 was given ten of these cysts on the

morning of July 24. Upon examination five and one-half hours

later, four young tapeworms (evaginated cysticerci) were found in

the posterior part of the duodenum attached to its wall. The re-

maining six cysts were in the stomach still in the invaginated con-

dition. Thus, it is obvious that the evagination of Cysticercus pisi-

formis occurs in the duodenum of the young cat.

Most of the tapeworms obtained in the experiment were young,

but two specimens had the reproductive organs fully formed. The

description here given includes the number and size of the ros-

tellar hooks of all of the tapeworms obtained (twenty-nine). Head,

0.846 mm. in diameter; rostellum powerful, armed with a double

row of 38 to 44 hooks, alternating large (225 to 270 microns) and

small (117 to 171 microns) ; ventral root of small hooks bifid. Neck,

1.75 mm. long, slightly narrower than the head. Genital pores irreg-

ularly alternate, very prominent. Proglottid at first narrow and

much shorter than broad, becoming nearly square in the posterior

region; testes numerous, filling interior of proglottid, but not -oc-

curring laterad of the excretory canals; vas deferens coiled before

reaching cirrus pouch; vagina leading to two ovaries in the distal

half of the proglottid; uterus single and median, not filled with

oncospheres. Length of strobila 22 mm. The lack of eggs, of

course, indicates that the specimens are not wholly mature, and

this accounts in part for their smaller size. All of the other char-

acters, however, agree so closely with those of T. pisiformis Bloch

that these tapeworms, apparently, are of that species.

The control kittens which were kept in the experimental cages,

but not fed cysticerci, did not contain a tapeworm, while seven of

the eight kittens that were fed Cysticercus pisiformis became infect-

ed with tapeworms, all of which agree with T. pisiformis in number

and size of rostellar hooks, and two larger specimens agree so closely

with this species that they are considered by the writers to be slightly

immature specimens of T. pisiformis Bloch.

CESTODE FROM THE YOUNG CAT 95

The larval form of this tapeworm develops occasionally in hares

and in mice (Stiles, 1906:43), but it is well known that its usual

intermediate host is the cottontail. Considering the abundance of

the latter and the large numbers of cats that have been examined

for parasitic worms, it is evident that T. pisiformis rarely develops

in the cat. However, this may be accounted for in part by the fact

that after the young cottontail has eaten the tapeworm oncospheres,

from six weeks to two months are required for the cysticerci to

develop and by this time the cottontails are usually large enough to

evade the cat.

This cestode is sufficiently generalized to develop also in the

fox (Cobbold, 1876 :674), and Benoit-Bazille and Dramard (1905:

10) report it (under the name of T. serrata Goeze) from the tiger.

On the other hand, evidences of specialization are not lacking. Sev-

eral attempts to infect man have failed according to Hall (1913 :43)

who ingested three cysticerci of this species with negative results,

and Scott’s (1913:263) attempt to infect swine likewise failed.

SUMMARY

The dog tapeworm, Tenia pisiformis Bloch, may develop in

the young cat (Felis domestica).

Evagination of Cysticercus pisiformis occurs in the duodenum

of the domestic kitten.

LITERATURE CITED

BenotT-BAzitte, H., and Dramarp, J.

1905. Deux nouveaux parasites du tigre royal. Naturaliste, (2) 28:10.

CopsoLp, Tu. S.

1876. Remarks on the Study of Parasites, with Suggestions in Reference

to the Management of Sheep suffering with Nematoid Worms.

Veterinarian, Lond., 49 :673-676.

Hatz, M. C.

1914. Experimental Ingestion by Man. of Cysticerci of Carnivore Tape-

worms. Jour. Parasit., 1:5-9,

96 J. E. ACKERT AND A. A. GRANT

Scorr, J. W.

1913. The Viability of Certain Cysticerci in Pigs and in Young Dogs.

Science, N. S., 37 :263.

Stites, C. W.

1906. Illustrated Key to the Cestode Parasites of Man. Bull. 25, Hyg.

Lab., U. S. Pub. Health and Mar.-Hosp. Serv., Washington,

pp. 1-104. |

NOTES, REVIEWS, ETC. 97

DEPARTMENT OF NOTES, REVIEWS, ETC.

It is the purpose, in this department, to present from time to time brief original

notes, both of methods of work and of results, by members of the Society. All

members are invited to submit such items. In the absence of these there will be given a

few brief abstracts of recent work of more general interest to students and teachers.

There will be no attempt to make these abstracts exhaustive. They will illustrate progress

without attempting to define it, and will thus give to the teacher current illustrations, and

to the isolated student suggestions of suitable fields of investigation.—[Editor.]

ENTOMOLOGICAL NOTES

Hemoglobin im Chironomus.—Leitch (716, Journ. Physiology,

50 :370-379) reports results of investigations on “The Function of

Hemoglobin in Invertebrates with special Reference to Planorbis

and Chironomus Larve.’ Hzmoglobin occurs in certain Chzto-

poda, Nemertea, Hirudinea, Gephyrea, Lamellibranchiata, Gastro-

poda, Brachiopoda, Ostracoda, Copepoda, the larve of Musca, and

the larve of Chironomus. Obviously, zoological kinship is not cor-

related with the presence of hemoglobin, but it appears that its

occurrence among invertebrates is correlated with a habitat defi-

ciency in oxygen. In connection with experiments to determine

the use and suggested storage function of hemoglobin, it was found

that the hemoglobin makes available “by its power of binding oxygen

chemically, a quantity of oxygen sufficient for the needs of the ani-

mals at oxygen tensions so low that the necessary amount is not

supplied by physical solution.” There is no storage of oxygen but

the action consists in the ‘‘constant binding of oxygen at the sur-

face of the body and the constant giving up of it in the interior:

a continuous mixing and interchange of oxidized and reduced blood

kept in motion by the beating of the heart.”

Reactions of Drosophila——Cole (717, Journ. Animal Behavoir,

7 :/1-80), in an experimental study of the reactions of Drosophila

ampelophia, finds that this insect, when creeping, shows negative

reaction to gravity, to centrifugal force which is equal to or slightly

greater than gravity, and to currents of air without regard to other

stimuli. It is shown that gravity is both a kinetic and a directive

98 AMERICAN MICROSCOPICAL SOCIETY

stimulus. Apparently, the stimuli causing these responses are re-

ceived by the sensory nerves of the leg muscles. However, it seems

probable that the flying reactions of this insect are not influenced

at all by gravity.

Insect Blood Cells in Vitro—Glaser (’17, Psyche, 24:1-6) has

experimented with the cultivation of insect blood cells in artificial

cultures. Using the caterpillars of Malacosoma americanum, Cwr-

phis unipuncta, Laphygma frugiperda, and Porthetria dispar as

sources of material, cultures were made of the corpuscles in their

own plasma, in Locke’s solution, or in a mixture of the two. Prep-

arations made from the second and third named species were kept

alive for over a month without washing out the cultures or trans-

ference to fresh media. Cultures of blood cells taken from the

last-mentioned species were kept alive for seventeen weeks with-

out washing or renewal of the media. In such cases, the cells were

no longer vigorous and exhibited signs of degeneration, but when

these old cultures were washed out and renewed with fresh media

the cells grew and multiplied as before. The ordinary amcebocytes

were the only structural elements in the blood which multiplied in

tissue cultures. Evidence leads to the conclusion “that the blood

cells, after their differentiation from the mesoderm during embry-

ological development, simply maintain their numerical equilibrium

in larve and adults by dividing mitotically at certain intervals.”

Development of Color Markings.—Onslow (’16, Biochemical

Journal, 10:26-30) has studied the development of the black mark-

ings on the wings of Pieris brassice and finds that they are caused

by the oxidation of a “colourless chromogen by a tyrosinase,” a

ferment which is supplied from the body-lymph of the pupa, “pos-

sibly by means of the wing-nervures,” to the chromogen which has

previously appeared in the regions destined to become black. The

form of the markings is determined by the particular position of

the chromogen. The oxidation occurs just previous to the emer-

gence of the imago and as soon as the oxygen of the air comes in

contact with the surface of the wing.

Effect of Cold on Malaria Parasites—King (717, Journ. Exp.

Medicine, 25:495-498) has investigated the validity of the older

assumption that the development of malaria parasites in Anopheles

NOTES, REVIEWS, ETC. 99

is arrested at a temperature of about 60 degrees F. and that the

parasites themselves are destroyed at lower temperatures. A series

of experiments with Plasmodium vivax (the parasite of tertian

malaria) in the mosquito Anopheles quadrimaculatus showed that

it can survive exposure to a temperature of 30 degrees F. for 2 days,

31 degrees F. for 4 days, and a mean temperature of 46 degrees F.

for 17 days. Ina smaller series of experiments with Plasmodium

falciparum (the estivo-autumnal parasite) in the same species of

mosquito, the sporonts showed a 24 hour resistance to temperatures

as low as 35 degrees F.

Nuclear Size in Nerve Cells—Smallwood and Phillips (16, ’

Journ. Comp. Neurology, 27:69-75) have studied the nuclear size

in the nerve cells of the antennal lobe of the honey-bee. Material

from twelve different stages of the life cycle showed a definite

variation in the size of the nuclei which cannot be explained as due

to the effect of fatigue or old age, but may be interpreted as “the

normal variation present in these cells.” It is thought that the

cells and their parts, such as nuclei, are the variable factors that

are responsible for the variation in the tissues and organs. “In

place of definite nuclear change with age, we find a constant varia-

tion which tends to rhythmic.”

‘Classification of Diptera—Malloch (17, Bull. Ill. State Lab.

Nat. Hist., 12:161-409) has recently published a very useful paper,

“A Preliminary Classification of Diptera, exclusive of Pupipara,

based upon Larval and Pupal Characters, with Keys to Imagines

in certain Families.” This paper is intended to meet the growing

need of taxonomic and economic entomologists, animal ecologists,

and others whose work would be facilitated by a synoptic treatment

of the immature stages of Diptera. It is rich in keys and descrip-

tions of the larval and pupal stages of species found in central Unit-

ed States, containing, in addition, concise information for the

different groups on habits, habitats, geographical distribution, and

important literature. Twenty-nine excellently executed plates in-

cluding three hundred seventy-three drawings of various life his-

tory stages and structural detail add much to the usefulness and

clearness of the paper.

100 AMERICAN MICROSCOPICAL SOCIETY rf

Biology of Fireflies —Williams (17, Journ. N. Y. Ent. Soc.,

25:11-33) reports: the results of an extended study on six species

of American fireflies, Photinus consanguineus, Photinus scintillans,

Ellychnia corrusca, Lucidota atra, Pyratomena sp., and Photurus »

pennsylvanica. Of these, the last named is the most brilliant and

the second named the faintest in luminescence in the adult stage.

The larval light producing organ in Photurus and Photinus differs

from the adult one which replaces it at maturity. The chief func-

tion of the light produced by adults is to bring the males and fe-

males together. The larve are all luminescent, the light appearing

at two points on the eighth abdominal sternite. Luminescence also

occurs in the pupal stage. Eggs are faintly luminous, at least for

a short time. The larve are probably all carnivorous, but among

the adults all seem to be herbivorous except the female of Photurus

which is carnivorous. In most cases, the larval life extends over

the greater part of two years. Ellychnia corrusca hibernates as an

adult, but all the others hibernate as larve.

PauL S. WELCH.

Kansas State Agricultural College.

NOTES ON OLIGOCH ATA

Development of Genital Organs.—Gatenby (’16, Quart. Journ.

Mier. Sci., 61:317-336) reports results of a study on “The Develop-

ment of the Sperm Duct, Oviduct, and Spermatheca in Tubifex

rivulorum.” The gonads are formed very early but not until the

worm is nearly half grown do the other parts of the genital organs

begin to appear and the ducts are not fully developed until some

of the genital products are ripe. The spermatheca begins to ap-

pear in X while the sperm duct is still incomplete and very rudi-

mentary. The oviduct is the last to be developed. “The more

highly differentiated structures begin to develop first, while the

oviduct, which is very simple, appears quite late.’ No satisfactory

evidence of dimorphism of spermatozoa, such as is reported by

Dixon (see below), was found. There is also disagreement with

Dixon’s statement that testes cannot be found in the adult because

they have been completely enclosed in the sperm sac, Gatenby hold-

NOTES, REVIEWS, ETC. 101

ing that the position of the testes is not affected in any way by

the formation of the sperm sac.

Tubifex.—Dixon (15, Proc. and Trans. Liverpool Biological

Society, L. M. B. C. Memoirs, No. XXIII, pp. 303-402) presents

results of an extensive morphological study of Tubifex. The ex-

ternal anatomy and all of the principal internal systems are described

in some detail, considerable attention being given to the reproduc-

tive organs. Among other things, the formation and transformation

of the sex cells are described and two distinct kinds of spermatozoa

are thought to have been demonstrated. A study of the parasites

of Tubifex showed the presence of the following internal parasites:

Urospora senuridis (gregarine) and Caryophylleus (cestode).

Opalina and Synactinomyxon tubificis, reported by other workers

in Tubifex, were not found. Certain fungi occur as external para-

sites, appearing, as a rule, in or near the setigerous sacs. Vorticelle

are frequently attached to the body-wall, but are not true parasites.

A bibliography of seventy-one titles and seven plates accompany

the paper.

PAauL S. WELCH.

Kansas State Agricultural College.

PARASITES IN THE MOUTH IN CASES OF PYORRHEA

Goodrich and Moseley (Jour. R. M. S., Dec. 1916) discuss

the parasites that are found accompanying pyorrhea. They hold

with Znamensky that pyorrhea always begins as an inflammation

of the margin of the gums, followed by gradual recession of the

gum. The tartar ridge at the margin of the gum is formed by the

bacterial Leptothrix which, if not regularly and completely removed

in the soft condition, becomes calcified and forms “hard” tartar.

The authors believe that the tartar is deposited chiefly or wholly by

Leptothrix, and note that excessive amount of tartar is an accom-

paniment of the disease.

They believe Leptothrix to be a pleomorphic organism appear-

ing in coccoid, fusiform, and filamentous form under different cir-

cumstances. The plant is found in all recessions and pockets of

the gums, in crypts of the tonsils, and may occur abundantly in

the bronchial tubes.

102 AMERICAN MICROSCOPICAL SOCIETY

Aside from Leptothrix the authors discuss Entame@ba gingivalis,

which a year or so ago was announced to be the cause of pyorrhea ;

Trichomonas hominis; and numerous bacteria, yeasts, and. some

spirochetes.

While not prepared to prove that any particular organism is

the primary cause of pyorrhea, the writers are disposed to incrim-

inate the Leptothrix.

Drew and Griffin (Jour. R. M. S., April 1917) by devising an

improved technic have been able to present the best microscopic pic-

ture yet published of the parasites of the mouth in pyorrhea. They

have studied systematically some 300 cases. The following are the

outstanding results: (1) amebe were found in every diseased

mouth, tho their numbers bear no relation to the severity of the dis-

ease, and were never found in normal mouths; (2) the same micro-

scopic population was found in all cases; (3) tartar is not always

found in diseased mouths ; (4) Leptothrix is invariably present; (5)

Trichomonas was found in 10% of the cases; (6) immense numbers

of spirochetes and treponemata, representing at least six species,

are characteristic; (7) bacteria and streptococci are invariably pres-

ent and often in immense numbers.

The authors do not feel ready to speak strongly about the

clinical aspects of pyorrhea. They think that the amebz have little

to do in causing the disease. They suggest that mechanical injury

plays an important part in the inception of the disease. When in-

jury once occurs, it is believed that the spirochetes play the chief

role in the disease by the destruction of tissue and the formation

of pockets. The ameboid cells are thought to destroy the red cells,

and the bacteria to form toxins. The authors claim that arsenic

preparation, such as atoxyl and salvarsan are of value in treatment,

that emetin is worthless, and that vaccine treatment is helpful.

The technic is as follows:

The material is removed from the pockets in the gums by a

Pasteur pipette whose fine extremity is drawn out to capillary thick-

ness. A centimeter of this is left on the pipette and turned at right

angles to it. A little 0.5% saline solution is allowed to enter this

fine extremity. This is then used as a probe in the pockets, whence

the infected material is drawn into the salt solution.

NOTES, REVIEWS, ETC. 103

For permanent mounts, a drop or two of this material well

mixed in the solution, is spread over a perfectly clean cover glass.

Then,— |

(1). Drop film side down in Schaudinn’s fluid for fixation.

(2). One half hour in each,—30, 50, 70, 80% alcohol to ab-

solute.

| (3). Pass back thru these grades, one half hour in each, to

distilled water.

(4). Place for 10 or 12 hours or more in 2% iron-alum so-

lution.

(5). Stain in 1% hematoxylin for 24 hours.

(6). Differentiate by immersing in 2% iron-alum, watching

the decolorization under the microscope until the nuclei are sharply

differentiated.

(7). Wash, pass thru grades of alcohol,—15 minutes in each,

—and clear in xylol. Mount in balsam.

FACTORS INFLUENCING THE SPORANGIAL CHARACTERS OF MYCETOZOA

A. E. Hilton (Jour. Q. M. C., Nov. 1916) gives a very sugges-

tive analysis of the factors whose interplay produces the interesting

variety we see in the sporangia of Myxomycetes. In some detail

he shows how surface tension, gravity, lateral compression, capil-

larity, desiccation, internal precipitation of solids,—and all the con-

ditions which modify any of these,—operate to produce the variety

of depressed, globular, cylindrical forms of these plants, with or

without stalks. It is a very good illustration of an intelligent effort

to show how “simple combinations of well known forces produce

complicated results.”

PROTOPLASMIC CONTINUITY IN EARLY EMBRYONIC DEVELOPMENT

Cameron and Gladstone (Jour. Anat. Physiol. Vol. 50: p. 207)

advance the view that the blastoderm of early animal embryos

does not show the cell demarcations which we habitually assume.

_ On the contrary the cytoplasm surrounding the nuclei is continuous,

and the nuclei themselves should be looked upon as the units. They

104 AMERICAN MICROSCOPICAL SOCIETY

regard the typical cellular structure as a derived and somewhat

degenerating process. Regarding the nucleus as central, both struc-

turally and functionally, the nascent endoplasm immediately sur-

rounding the nucleus is derived from the nucleus which has manu-

factured it from the food taken in by the cytoplasm. The nascent

endoplasm is gradually transformed into the outer maturer endo-

plasm, and this gives rise in turn to the ectoplasm. The functions

are less and less active, passing from nucleus to ectoplasm. The

authors therefore look upon the early embryo as a differentiating

plasmodium. They claim to have traced this condition into the

three-layered stage in vertebrate embryos.

FACTORS CONTROLLING THE RATE OF REGENERATION

Zeleny (Ill. Biol. Monog., Aug. 1916) continues his studies

on regeneration and the factors, internal and external, that may

influence it. In this monograph of 170 pages the author investigates

the following points upon amphibian larvae :—

1. Rate of regeneration from new tissue compared with that

from old tissue. The general conclusion is that the rate of regen-

eration is independent of the age of the cells near the cut surface,

except in those early stages where cell migration rather than cell

division secures the regeneration, in which case the rate of regener-

tion may be greater from new tissue.

2. Rate of regeneration as determined by successive removal.

The rate of successive regeneration is found to decrease with suc-

cessive removals in the same individual. The factor of age of

course enters into such a case. By eliminating the age factor the

investigator found that there is no decrease in rate for the second

and third regenerations. Indeed the second has an advantage over

the first; and the third somewhat less over the second.

3. The effect of the level of the cut on the rate and complete-

ness of regeneration. In general it was found that the rate of

regeneration varies directly with the amount of material cut away,—

the deeper the cut the more the regeneration both in rate and

amount. But in any event the regeneration stops short of com-

plete replacement.

NOTES, REVIEWS, ETC. 105

4. Effects of degree of injury. Within reasonable limits re-

generation of a part is not retarded by the simultaneous removal

of other and different parts. The removal of similar parts may

even accelerate the regeneration; e. g., the right foreleg regenerates

more rapidly when the other foreleg is also removed than when the

right alone is removed.

The author finds that the rate of regeneration varies,—starting

slowly, increasing rapidly until near its maximum, then decreases

rapidly, and finally decreases slowly to zero. The forces that cause

the cessation of regeneration seem to stop the process short of com-

plete regeneration.

THE HEAD AND MOUTH PARTS OF DIPTERA

Peterson (Ill. Biol. Monog. III:2, Oct. 1916) presents the re-

sults of a study of 53 of the 59 families of North American Diptera.

Twenty-five plates with more than 600 figures accompany the mono-

graph.

The plan consists in constructing a “hypothetical type” for the

head capsule and each of the mouth parts, with which to compare the

particular forms. This hypothetical type is formed by consideration

of the parts of generalized groups of insects and of the less special-

ized conditions in the Diptera themselves. All the different parts

are brought into comparison with this hypothetical type as well as

with one another.

Modification of the fixed and movable parts from this general-

ized type usually take the form of reduction, change of shape, loss of

chitinization, or expansion of the membranous areas.

The results are too technical and of too much detail to report in

brief space and apart from the figures. While the mouth parts show

wide modifications in the order, all of them, including the epipharynx

and the hypopharynx, retain their relative position,—although they

are sometimes extruded a considerable distance from the head

capsule.

The compound eyes are unusually well developed. They show

secondary sexual characters in a larger number of species than do

any other of the head parts.

106 AMERICAN MICROSCOPICAL SOCIETY

EARLY CASTRATION OF THE VERTEBRATE EMBRYO

Reagan in a paper before the American Society of Zoologists,

after reviewing the various suggestions as to the place of origin of

the germ cells of the vertebrate embryo, supports the contention

of Swift that in birds the primordial germ cells are originated in a

crescentic area of entoderm very early in the chick embryo, then

enter the mesoderm which later invades the region, thence enter

the blood stream by way of the blood vessels forming in the meso-

derm, and migrate from the base of the mesentery into the position

of the gonad.

By cutting out the crescentic area very early Reagan found that

sex cells were not to be seen in the gonads at an age when in normal

individuals many large sex cells are present in them.

The writer believes that this early castration tends to show that

the only primary sexual characters are the sex cells themselves, and

that even such organs as the efferent ducts are secondary, dependent

on secretions from the gonad. If these secretions are from the

interstitial cells rather than the sex cells, it seems that the presence

of the sex cells themselves, in the gonad, is necessary in order to in-

duce it.

THE SEGREGATION AND RECOMBINATION OF CHROMOSOMES

Corothers (Proc. Am. Soc. Zool. 1916:22) reports on the

behavior of the chromosomes in two species of short horned grass-

hoppers. Because of the double fact (1) of constancy in the size,

shape, and number of the homologous chromosomes in each indiv-

idual, and (2) of variations in the homologous chromosomes in dif-

ferent individuals resulting in certain constant differences (the latter

fact not having been reported before), she finds that the segregation

of the chromosomes may be traced in the gametes derived from both

parents. This is found to take place according to the law of chance.

A further study shows their recombination in such complexes as

would result from random unions and as would be demanded in

Mendelian inheritance.

NOTES, REVIEWS, ETC. 107

LAYING CYCLES IN BIRDS

Cole (Proc. Am. Soc. Zool. 1916: p. 32) reports on two distinct

conditions shown in the cycle of egg-laying in birds. In one the

number of eggs is definite and predetermined when laying begins;

in the other it is indeterminate and the cessation of laying and

beginning of brooding is determined by stimuli that operate during

the process of laying. In the latter case the number of eggs in the

nest operates as such a stimulus. If the eggs are removed as

deposited the period will be lengthened. The pigeon and English

sparrow illustrate the determinate, and the common fowl and the

house wren the indeterminate cycle.

THE PINEAL GLAND AND PIGMENTATION

McCord and Allen (Jour. Exp. Zool., May 1917) give the results

of experiments upon tadpoles through feeding upon pineal gland

substance. It was found that such feeding had no effect upon pig-

mentation up to the tenth day. From this point to the end of meta-

morphosis animals fed upon pineal food, or absorbing acetone ex-

tractives from pineal gland. through the gills and skin, pass through

a striking cycle of pigment changes. Soon after feeding they begin

to lose color and within thirty minutes all macroscopic appearance of

pigment is lost. Unless more pineal substance is added complete

restofation of color occurs in three to six hours.

Microscopic examination shows that the change is due to the

aggregation of the pigment in the center of the large, branching,

moss-like melanophores of the sub-epithelial tissues. It is found

also that the pineal extracts cause contraction in smooth muscle

fibres as well as in melanophores.

The theoretical bearing of’ these experiments comes from the

view,—as is shown in some lizards,—that the original pineal body

had an optical function. The possible adaptive value of pigment de-

pends upon light. Eyes are essential controlling factors in the

adjustment of color to the environment. But in many types there

are still pigment changes even in blinded specimens. Theoretically

the pineal body might retain enough of its light sensitiveness to be

stimulated and thus to exert an influence upon the pigment cells.

108 AMERICAN MICROSCOPICAL SOCIETY

If not, it nevertheless secretes materials which may produce pig-

ment changes independent of the conditions of the environment.

These facts suggest explanation of the somewhat diverse and

even at times discordant behavior of pigments.

COOPERATIVE TECHNIQUE

McClung (Anat. Rec. 12:3, April 1917) suggests the need and

value of combination and cooperation in respect to microscopical re-

search, similar to that which has proved so effective in business. To

this end he proposes a concerted series of studies upon the technical

processes as such, by people prepared to extend these over a wide

range of research. These results would be reduced to the most com-

pact form and published together. In addition there might be built

up at some central place a collection of actual preparations illustrat-

ing the use of these methods, each the work of an expert in the

particular field. These slides (or other form of object) might be

subject to loan to other microscopic anatomists.

FIXATION OF MAMMALIAN CHROMOSOMES

Hance (Anat. Rec. 12:3, April 1, 1917) reports studies illustra-

tive of the view-point of Professor McClung in the preceding ab-

stract. His task was to find processes that would give best results

in revealing chromosome conditions in mammals. Two desiderata

are held in view:—the general fixation must be such as to avoid

shrinkage and distortion; and the method must so far as possible

lead to distinctness, differentiation, and separateness of the chromo-

somes in the various stages.

The method most fully meeting these ends is outlined as fol-

lows:

1. To be sure of getting one or more specimens in a “cycle of

division,” material should be taken from as many animals as pos-

sible.

2. Place small or finely teased pieces of fresh tissue immedi-

ately into cold Flemming’s solution, plus urea. This should act

20-24, or more, hours.

NOTES, REVIEWS, ETC. 109

3. If the above method does not succeed, allow the small pieces

of fresh tissue to remain exposed to the action of the air for ten

to twenty minutes before placing in the Flemming solution. This

should be at a temperature of 4° C. Time as in 2.

4. Wash in water about 24 hours.

5. Dehydrate by very gradual steps.

6. Clear from 95 per cent alcohol in cedar oil followed by xylol.

7. Imbed in paraffin.

QUINCE JELLY AS A CULTURE MEDIUM FOR EUGLENA

Turner (Anat. Rec., 12:3, April, 1917) develops a mode of

culturing Euglena and other green protozoa in quince jelly, which

has practical value for the general laboratory because of the ease

of making and preserving the cultures. Cultures have been kept for

more than a-year, and may be successfully transplanted.

The methods and results may be summarized as follows:

1. Twenty grams of dry quince seed boiled for half an hour

in 1.5 liters of distilled water produces enough of the exudate, aiter

the seeds are strained out, to make a total culture mass of 2% liters,

by the addition of distilled water.

2. A still more fruitful medium was made by grinding the

seeds to a fine powder or meal and leaving it in the jelly. Water to

21% or 3 liters may be used with this amount of meal. This medium

has the disadvantage of being opaque.

3. Cultures develop more rapidly in a slightly thinner medium,

but more lasting cultures call for a thicker medium.

4. The medium should be made slightly alkaline, and the

cultures kept at room temperature, in moderate light.

5. Test tubes, vials, and similar vessels furnishing large ver-

tical range and little exposed surface are best.

6. The reactions of Euglena are modified in interesting ways,

as compared with their usual behavior in water.

“VITAL DYES” IN TELEOSTS

Wislocki (Anat. Record. XII, 4, May 1917) reports on experi-

ments with benzidine dyes injected into the peritoneal cavity of

110 AMERICAN MICROSCOPICAL SOCIETY

teleosts. These dye-stuffs form ultra-microscopic solutions which

are non-toxic and are taken up and housed by various mono-nuclear

cells. Trypan blue in a few hours shows as a pale blue in all body

tissues except the central nervous system and fatty tissues. The

particulate storage of the dye does not show until the third day and

onward, reaching a maximum in about a week.

The building and storage (phagocytizing) of the dyes in teleosts

is seen in the endothelium of the hepatic sinuses ; by the reticulo-

endothelium of the spleen; the endothelium of the renal portal sys-

tem (although it does not occur in the Wolffian body of amphibia

nor in the metanephros) ; and in the endothelium of the lymphatic

vessels.

ACID COLLOIDAL DYES IN BLOOD AND TISSUE CELLS

Downey (Anat. Rec., XII:4, May 1917) holds that the ability

to ingest and store the colloidal dyes is not diagnostic, as has been

thought, in distinguishing between phagocytes of tissue origin and

those that have come from the blood stream. He believes that their

state of differentiation and the special conditions under which they

happened to exist at the time of experiment, and not their origin, de-

termines their reaction to the dye. Any phagocyte may store the dye.

He agrees that the coloring is merely a process of ingestion and

storage, and not in any sense a real staining of preformed structures.

GENETICS AND EUGENICS

No department of Biology has in recent times more appealed

to the popular imagination or has been more widely and _ thoroly

treated by capable writers than that of inheritance. This is not

surprising when we recall the great importance of heredity to the

whole philosophy of life and further realize that more progress

has been made in clearing up and organizing the phenomena of

inheritance in the last 25 or 30 years than in all the rest of human

history put together. The coincidence of this clearer knowledge

of genetics with several movements for human betterment has in-

sured that the application of these newly discovered principles of

animal and plant breeding should be made to human beings,—at

NOTES, REVIEWS, ETC. 111

least in theory. In this way Eugenics has seized the human imag-

ination,—Eugenics meaning “The study of agencies under social

control that may improve or impair the racial qualities of future

generations, either physically or mentally.”

Professor Castle, after developing somewhat historically the

contributions of various investigators to our ideas of evolution and

heredity,—including Darwin, Lamarck, Weismann, Spencer,—dis-

cusses the great movements of biometry and experimental breeding

and their contribution to the fundamental problems. His treatment

differs from that found in the similar books largely in the omission of

the customary diagrams and formule; in a larger introduction of

photographs illustrating animal genetics ; in furnishing numerous ta-

bles giving the unit characters which have been subject to experiment

in various plants and animals, together with the fact and degree of

dominance in these characters as revealed by the experiments; in

organizing the presentation around the discoveries themselves rather

than in accordance with a priori analysis of the subject matter, and

thus giving it a rather more objective and concrete character.

The section on Eugenics includes chapters on Human crosses;

Physical and mental inheritance in man; Heredity of general men-

tal ability, insanity, feeble-mindedness; and the Possibility and

prospects of breeding a better human race.

The book concludes with an appendix containing a translation

of Mendel’s original paper—‘‘Experiments in Plant-hybridization” ;

and an extended bibliography.

The author intends the book as a text book for college stu-

dents, as a reference book for breeders of plants and animals, and

as a summary of present views for the intelligent person interested

in this field of research. In the opinion of the reviewer the book

is well calculated to succeed in meeting these purposes.

Genetics and Eugenics, by W. E. Castle. Illustrated; 346 pages. Harvard Uni-

versity Press, Cambridge, Mass. 1916. Price, $2.00, postpaid.

112 AMERICAN MICROSCOPICAL SOCIETY

SPENCER-TOLLES FUND: REPORT FOR 1916

Owing to the failure to hold the annual meeting for 1916 the following

report of the Custodian has not been acted upon. It is published now that

the Society may realize that the $5000 goal has been reached. The year 1916

saw the largest annual addition ever made to the fund. It is the hope and

expectation that the income of the Fund will from this time forward: be

made effective in advancing the interests of microscopic research, and thus

reinforcing the work of the Society. Only by so doing can we make the

Fund a real memorial to these early pioneers of microscopy.

Reported at Columbus meeting ............. $4489.32

Dividends PECEived’ biecu4 Fade lene be fon ts See $ 416.22

Shed OLN. CANSACLIONS LUV aaa ss oie eealne Ae 120.00 536.22

$5025.54

GrAND TOTALS

All contributions to date ........ccceeecenee .$ 800.27

All sales iol) PAnSaChONnS 5) uséh cn sagie ge ta ee 878.38

All ife-memberships Ui) sc even vecasveuwwy wep 300.00

All interest and dividends ................00- . 3236.89 $5215.54

LESS

All grants: to’ date -4)\ 25 sas Vives preneaeenas $ 150.00

All life-membership dues paid ........-.+2+45 40.00 190.00

$5025.54

Life-members: (Robert Brown, dec’d.) ; J. Stanford Brown; Henry B.

Duncanson; A. H. Elliott; John Hately; Seth B. Capp.

Contributions of $50 and over: John Aspinwall; Iron City Micro-

scopical Society; Magnum Pflaum; Troy Scientific Society.

Macnus Priaum, Custodian.

CHARLES EDWARD HANAMAN

NECROLOGY

Charles Edward Hanaman was born in Port Schuyler near

Watervliet, Albany County, New York, on Noveniber 19, 1848.

He was educated at St. Paul’s School in Troy and at the Troy

Academy, and expected after graduation at the latter institution to

attend college. His father had large interests in the milling indus-

try and wanted the assistance of his son in the business so much

that the boy surrendered his idea of a college career and em-

barked upon active business life. Those who knew Mr. Hanaman

well were always conscious of the real sacrifice that he made, but

they were equally convinced that he never expressed any real regret

at the change of plan. His interests in mercantile life were sur-

rendered in 1888 in order that he might assume the position of

Secretary and Treasurer of the Troy Savings Bank, and after serv-

ing four years in that capacity he was elected President of the bank,

a position which he retained continuously until his death which oc-

curred on September 2, 1916.

In 1900 he was selected as third Vice-President and member

of the Executive Committee of the Savings Bank Association of

New York state. Later he was twice elected President of the same

Association. His business ability was utilized in other directions

also for the benefit of the public as he served for twenty years

as Treasurer and for five years as Vice-President of the Troy

Orphan Asylum. He was also for twenty years Treasurer of St.

John’s Episcopal Church. In all of these capacities he served the

community not only faithfully but with marked ability, and the

different institutions which enjoyed his services bear lasting evi-

dence of his integrity and efficiency as‘a financial leader.

His interest in scientific things was developed at an early age.

It is recorded that when nine years old he started out with the

use of the microscope and became absorbed in its revelations so

that he continued work with it throughout his entire life. He found

in this study his greatest pleasure and an absorbing interest which

led him to devote every spare moment to it. For his own use he

built up in the course of years a splendid collection of the most

114 AMERICAN MICROSCOPICAL SOCIETY

varied instruments of the biological laboratory and tried out per-

sonally all, or nearly all, of the methods that have been developed

within the last fifty years for the preparation of materials and the

investigation of the microscopic structures of organisms. His large

cabinet of prepared specimens was opened with delight for any who

expressed the slightest interest and even the technically trained

man found in it something that well repaid the time spent in an

examination of the materials.

In spite of the insistent demands of a busy commercial life he

subscribed to many leading biological journals and perused them

so as to become thoroughly familiar with their contents. He de-

lighted in nothing more than to review current theories and recent

discussions of the most technical type with those familiar with such

subjects, who crossed his path and in such discussions displayed

a clearness and breadth of knowledge that was as surprising as

unusual in a man of affairs. He was one of the early members

and officers of the Troy Microscopical Society and a charter mem-

ber of the American Society of Microscopists, the predecessor of

the American Microscopical Society. He was also a Fellow of

the Royal Microscopical Society of London and the American Asso-

ciation for the Advancement of Science, and a member of the

Biological Society of Washington.

Mr. Hanaman was a splendid sample of a type rare in this

country, for although prominent and in marked degree successful

in business life he had an avocation in his scientific studies which

he pursued with equal zeal and success. He will long be remem-

bered as one who stimulated and encouraged by his interest and

vigor the scientific workers with whom he came in contact. Science

needs a multitude of such men and owes a great debt of gratitude to

the few who in this way gain interest in its work and contribute in

many ways to the support and extension of that work.

Personally Mr. Hanaman was quiet and unobtrusive so that

few except those who were brought in immediate contact with him

knew of his thorough knowledge of biological investigation and of

his personal interest in that field. In his death the American

Microscopical Society lost a warm friend and an active supporter.

NOTEWORTHY BOOKS

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ORGANIZED 1878 INCORPORATED 1891

VOLUME XXXVI

TRANSACTIONS

OF THE

AMERICAN

MICROSCOPICAL

SOCIETY

PUBLISHED QUARTERLY BY THE

AMERICAN MICROSCOPICAL SOCIETY

DECATUR, ILLINOIS

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Attention of members of the

AMERICAN MICROSCOPICAL SOCIETY

is called to the announcement of Grants

from the Spencer-Tolles Research Fund.

(See TRANSACTIONS, July, 1913)

These awards are to stimulate and aid research in

Microscopic work

1. The Grantee must be a member of the Society

2. Resulting discoveries must first be offered for publi-

cation in the Society Transactions.

3. Application should be made to Dr. Henry B, Ward,

Chm., Urbana. IIl.

NOTICE TO MEMBERS

It is a source of regret to the Editor that the Transactions can-

not be issued as dated. The issue is dependent, however, upon the

income of material from the members, and must await upon a certain

degree of balance in the material as well as quantity of material.

The Secretary will consider it a favor if members will notify

him of non-receipt of numbers of the Transactions, and of changes

of address.

| T. W. GALLoway,

Secretary-Editor.

Beloit, Wisconsin.

TRANSACTIONS

OF THE

American Microscopical

Society

ORGANIZED 1878 INCORPORATED 1891

PUBLISHED QUARTERLY

BY THE SOCIETY

EDITED BY THE SECRETARY

VOLUME XXXVI

NuMBER THREE

Entered as Second-class Matter December 12, 1910, at the Post-office at Decatur,

Ilinois, under act of March 3, 1879.

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Ree RPO adel VVC COALIOWAY «6570, Vidub ecw cwsledid wan iecere tual Beloit, Wis.

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EX-OFFICIO MEMBERS OF THE EXECUTIVE COMMITTEE

Past Presidents Still Retaining Membership in Society

R. H. Warp, M.D., F.R.M.S., of Troy, N. Y.,

at Indianapolis, Ind., 1878, and at Buffalo, N. Y., 1879

Apert McCa ta, Ph.D., F. R. M. S., of Chicago, Ill.

Gro. E. Fert, M.D., F.R.M.S., of Buffalo, N. Y.,

at Chicago, IIl., 1883

at Detroit, Mich., 1890

Suwon Henry Gace, B.S., of Ithaca, N. Y.,

at Ithaca, N. Y., 1895 and 1906

A. Cuirrorp Mercer, M.D., F.R.M.S., of Syracuse, N. Y.,

at Pittsburg, Pa., 1896

A. M. Brerte, M.D., of Columbus, Ohio,

at New York City, 1900

C. H. E1cENMANN, Ph.D., of Bloomington, Ind.,

at Denver, Colo., 1901

E. A. Birce, LL.D., of Madison, Wis.

at Winona Lake, Ind., 1903

Henry B. Warp, A.M., Ph.D., of Urbana, III,

at Sandusky, Ohio, 1905

HERBERT Ospzorn, M.S., of Columbus, Ohio,

at Minneapolis, Minn., 1910

A. E. Hertzier, M.D., of Kansas City, Mo.,

at Washington, D. C., 1911

F. D. Heap, Ph.D., of Philadelphia, Pa.,

at Cleveland, Ohio, 1912

CuHarLeEs Brooxover, Pu. D., of Little Rock, Ark.,

at Philadelphia, Pa., 1914

Cuartes A. Kororp, Ph.D., of Berkeley, Calif.,

at Columbus, Ohio, 1915

The Society does not hold itself responsible for the opinions expressed

by members in its published Transactions unless endorsed by special vote.

TABLE OF CONTENTS

FOR VOLUME XXXVI, Number 3, July, 1917

The Enchytreidz (Oligocheta) of the Woods Hole Region, by Paul

SC RAR ION ba Ss Diu tha A bia aiblela etal pie ule Ge. e’are.o fia’ ® Wisin cm pee hip bonis CNTs We

Chemical Microscopy, with Plates XII-XV, by E. M. Chamot............

Some Comparisons Between Nuclei of Nerve Cells, with Plate XVI, by

WA Eikiteres A ETSI R ra 3s 51k aR A Ce cbs Bk le eh 4 ba nica bi eolee bh bees

Notes and Reviews: Culturing of Microscopic Organisms for the

Zoological Laboratory, by George R. LaRue; Effects of Thyroid on

Paramecium; Are Conjugation and Encystment Necessary? Extra

Contractile Vacuoles in Paramecium; Preserving Fish without Ice;

Cytological Changes Accompanying Desiccation in Rotifers; The

Fundus Oculi of Birds, by Dr. Casey A. Wood.......-..---0 eee eeees

(This Number was issued on November 5, 1917.)

119

139

157

TRANSACTIONS

American Microscopical Society

(Published in Quarterly Installments)

per aren on a ena era eps eyepiece ienipones tthe)

Vol. XXXVI JULY, 1917 No. 3

THE ENCHYTRAEIDZ (OLIGOCHZTA) OF THE WOODS

HOLE REGION, MASS.*

By Paut S. WeEtcH

PAGE

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INTRODUCTION

During the summers of 1915-16, the writer carried on some

experimental studies with certain Enchytreide (Oligocheta) at

*Contribution from the Entomological Laboratory, Kansas State Agricultural College,

No. 29.

120 PAUL S. WELCH

Woods Hole, Mass. Owing to the fact that the known marine spe-

cies of that locality intermingle in the same habitats, and that some

of them have been reported from both fresh-water and terrestial

conditions, it was necessary to identify the enchytraids from the

various collecting grounds about Woods Hole in order to avoid

error in handling immature individuals and to insure accuracy in the

identity of the materials used. In spite of the fact that two species

occur in great abundance in the environs of Woods Hole and certain

other parts of the Atlantic Coast, they have received scant attention

from workers in any of the fields of zoology.

The writer wishes to acknowledge indebtedness to Director F.

R. Lillie for the privilege of using the facilities of the Marine Biolog-

ical Laboratory during the time this and other related work was in

progress.

ENCHYTRZUS ALBIDUS HENLE

Previous Records

Four records of this species on the Atlantic Coast occur in the

literature. Verrill (73) was the first to discover it in North Amer-

ica, describing it as Halodrilus littoralis, but its identity with Enchy-

treus albidus Henle was later established by Michaelsen (’00, p. 89).

Verrill reported it from “New Haven; Woods Hole; Casco Bay,

Maine; very common under dead sea-weeds and stones near high-

water mark.” Smith (95) made a careful anatomical study of this

annelid at Woods Hole, using the name Enchytreus littoralis Verrill.

He found the specimens extremely abundant in the same kinds of sit-

uations described by Verrill. Moore (’05) reported Enchytreus al-

bidus very numerous at Woods Hole as well as along the coast from

Casco Bay, Maine, to Sea Isle City, New Jersey. He not only found

them in the inter-tides zone of the shore but also “in moist spots

on farm lands on Martha’s Vineyard, where it could readily be intro-

duced in the large quantities of eel grass that are annually spread for

fertilizer.” He likewise states that “about Woods Hole it also lives

in damp, sandy woods and on the shores of fresh-water ponds, espe-

cially of one that formerly was connected with the Sound.” Passing

notice is made of all of the above-mentioned records by Sumner, Os-

THE ENCHYTR2IDZ OF THE WOODS HOLE REGION 121

burn, and Cole (713). Hunt (715) collected this species “in abund-

ance from the coarse gravel of the tidal zone on the seashore at Cold

Spring Harbor, Long Island, New York.”

HABITAT

Specimens of £. albidus were found in great quantities in sev-

eral of the protected coves about Woods Hole where the tide casts

up masses of eel-grass on sandy or gravelly beaches. They appear to

be restricted entirely to the inter-tides zone since no specimens were

collected in the comparatively dry sand above the high tide line and

none was observed beyond the low tide limit where the sand is con-

stantly covered with water. At low tide, the windrows of eel-grass

retain a considerable amount of moisture, thus keeping the under-

lying sand saturated and protecting it from the heat, light, and dry-

ing effect of the summer sun. At high tide, the entire zone occupied

by these worms is submerged. They were absent from sandy beaches

which lacked the masses of eel-grass, and even when the latter were

present, the worms were apparently much reduced in numbers when

the sand contained little or no organic debris. In July and August,

these enchytrzids occur in great numbers on the beach just opposite

the Lecture Hall of the Marine Biological Laboratory. The princi-

pal features of this habitat are a smooth, sandy-gravelly, gently slop-

ing beach; thick layers of eel-grass on the inter-tides zone at low

tide; quantities of fine pieces of coal and cinders mixed with the

sand; large amounts of organic matter in various stages of decom-

position; and complete protection from wave action. The worms

have a rather definite vertical distribution. In the matted eel-grass,

they usually occur in very small numbers, but appear in large quan-

tities in the first one to three inches of underlying sand and gravel.

Below these depths, the number decreases very rapidly. They show

a distinct’ tendency to accumulate about small rocks and pebbles

which are imbedded in the sand so that when such an object is

removed, the walls of the cavity are frequently covered with worms.

Several examinations of the various brackish, fresh-water, and

moist woodland habitats did not reveal the presence of these en-

chytrzids, although Moore (’05, pp. 394-395) reported them in such

places. However, it is very possible that a more thorough search

122 PAUL S. WELCH

would reveal them under the above-mentioned conditions, since in

addition to Moore’s record they are definitely known from such

habitats in other parts of the world.

METHODS OF CULTURE

Specimens were easily kept alive in the laboratory by placing

a number of them in shallow stender dishes containing sea water to

the depth of about one centimeter, to which had been added pieces

of fresh, green eel-grass from the drift on shore. In most of the cul-

tures used by the writer, the eel-grass was partly cleaned by washing

and wiping each blade, in order to avoid introducing into the culture

an excess amount of the extraneous matter adhering to the eel-grass

which interfered with the examination of the cultures for cocoons.

All of the cultures were kept covered, some being kept in darkness

while others stood in diffuse sunlight. In such cultures, the speci-

mens were active, cocoons were produced, and the mortality was

very low, the worms living until the cultures were purposely de-

stroyed, in some cases, after thirty days. Should it be desirable, the

water in the cultures can be reduced to the amount necessary only to

keep the worms and eel-grass thoroughly moist. Hunt (15, pp. 496,

501, 503) cultured this same enchytreid for regeneration expert-

ments by placing them “in small sterilized glass bottles, each contain-

ing a strip of filter paper and enough sterilized sea water to keep

the animals well moistened.” He also found that the worms could

be kept in such bottles moistened with “fresh water” in which they

regenerated as well as in the salt-water environment. It would ap-

pear from Hunt’s statement of methods that his specimens were not

submerged and he makes no statement as to the source of the “fresh

water.” The writer was unable to rear these enchytrzeids at Woods

Hole in the fresh water taken from the laboratory supply. In no

case did the worms, when submerged, live more than eighteen hours.

However, the addition of a very small amount of sea water to the

fresh water made the culture a fairly favorable one.

THE ENCHYTRZIDZ OF THE WOODS HOLE REGION 123

LuMBRICILLUS LINEATUS ( Mill.)

Identity

The final decision as to the identity of this annelid was reached

after the completion of a detailed study of the anatomy and a thor-

oughgoing examination of the literature dealing with a number of

European species which seem to exhibit various degrees of mutual

affinity. The complex character of the identification was due to the

existence of a number of very similar descriptions of forms under

several different names. The writer believes that this study of

Woods Hole material has helped to solve some of the difficulties and

to establish the identity of certain so-called species. While access to

the European material has not been possible, the descriptions, discus-

sions, and figures of the morphology of these foreign forms are suf-

ficiently detailed to serve as a working basis for comparison.

The anatomy of these worms agrees very closely with the de-

scription by Moore (’05, pp. 395-397) of a form which he collected

at Woods Hole and named Lumbricillus agilis. The only points of

disagreement have to do with the position of the clitellum, the posi-

tion and number of the ventral glands, and the character of the sper-

mathecal glands. These matters will be discussed later under their

respective headings and the apparent discrepancy accounted for, at

least in large part. It will suffice to state here that there seem to

be no grounds for a possible suggestion that the material studied by

Moore represented a species different from the one examined by the

writer.

An inspection of the descriptions of other species ascribed to

Lumbricillus reveals at least four foreign ones which, as described,

approach the Woods Hole form too closely to be disregarded, namely,

litoreus (Hesse), lineatus (Mull.), subterraneus (Vejd.), and verru-

cosus (Clap.). That some of the above-mentioned species are not

valid has been suspected before. Southern (’09, p. 150) pointed out

that the differences between /itoreus and lineatus seemed of no great

importance and that it “is doubtful whether there is sufficient justifi-

cation for keeping the two species apart” and Michaelsen (’O1 a, p.

207) has indicated the probability that /imeatus and subterraneus

should not be separated. Friend (16, pp. 23-24), in the examination

124 PAUL S. WELCH

of specimens from the sewage works of Manchester, England, found

that “every intermediate stage” between verrucosus (Clap.) and

lineatus could be detected. With respect to verrucosus, the only

points of distinction of any importance between it and lineatus have ©

been the number of sete per bundle and character of the ventral

glands. A careful parallel column comparison of the characters of

all the above-mentioned enchytrzids, together with those of the

Woods Hole specimens, shows that the range of variability of the

body length, setz, origin of the dorsal blood-vessel, shape of the mar-

gins of the brain, dimensions of the spermiducal funnel, and number

and development of the ventral glands are such as to link all of

these so-called species together. A detailed comparison is out of

place here, but reference to the earlier descriptions and a comparison

with the following discussion of the morphology of the Woods Hole

material will bear out this conclusion.

Since L. litoreus (Hesse), L. subterraneus (Vejd.), L. lineatus

(Miull.), L. verrucosus (Clap.), and L. agilis Moore must all be re-

garded as the same species, the question arises as to what name shall

persist to designate it. It appears that lineatus (Miull.) (1771) has

priority, but Ditlevsen (’04, p. 431) holds that lineatus should give

place to Levinsen’s name rivalis (1884): “Michaelsen hat gewiss

recht zu sagen, dass die Art von Levinsen dieselbe wie Pachydrilus

germanicus Mich. ist; aber nichts in der Beschreibung von O. Fr.

Miiller berechtigt ihn dazu, diese mit Lumbricus lineatus O. Fr.

Miller zu identifizieren. Die Charaktere, welche O. Fr. Miller

angibt, passen nur gemeinsam auf die Gattung Pachydrilus; es ist

aber durchaus unmoglich einige ihrer Arten danach zu bestimmen.

Da Levinsens Beschreibung die alteste ist, muss die Art den

Namen Pachydrilus rivalis behalten, und die Art Michaelsens

muss dieser zugeordnet werden.” The writer has examined the

description by O. F. Miller and its vagueness and lack of specific

detail is certainly apparent, giving ample reason for the numerous

attempts to place this uncertain “Lumbricus lineatus.’ The descrip-

tion might apply to almost any of the species assigned to Lumbri-

cillus. On such grounds it might appear that Ditlevsen’s contention

has some basis but it is evident that, if the synonymy proposed in

this paper is correct, the name could not be rivalis Levinsen since the

THE ENCHYTRZIDE OF THE WOODS HOLE REGION 125

name verrucosus has priority over it. At the present time there

seem to be grounds for regarding verrucosus as the valid name, but

since some uncertainty still exists concerning some of the other

apparently closely related species, the writer prefers to retain, for

the present, the old name of /imeatus.

There is growing evidence that an overindulgence in “species

making” has occurred in the genus Lumbricillus and the writer is

convinced that at least two other so-called species, and probably

more, are the same as lineatus. The genus needs extensive revision

and, when worked over in connection with complete and dependable

morphological data, it is very probable that the number of species

which the genus is now supposed to contain will be materially re-

duced.

With respect to the geographical distribution, these annelids

have been reported under the old names as follows: L. lineatus:

Denmark, Germany, North Russia, and England. L. litoreus: Italy

and Ireland. L. verrucosus: Hebrides, Terra del Fuego, Switzer-

land, Ireland, Scotland, and England. JL. subterraneus: England,

Ireland, Scotland, France, and Bohemia. L. agilis: Casco Bay to |

Vineyard Sound, Mass. Evidently this species is a cosmopolitan

form of wide distribution.

HABITAT

In connection with the above rearrangement, it is interesting to

consider, under the old names, the habitats from which these animals

have been reported. JL. lineatus has been recorded from debris and

under stones on the seashore, or river margins, and on aquatic plants

in fresh-water ditches. L. litoreus was reported on the seashore

under stones. L. verrucosus has been found on the seashore under

stones, in fresh-water situations, and, with some uncertainty, from

earth near manure heaps. L. subterraneus has been collected in

springs and aqueducts, in sewage works, in streams contaminated

with wastes, and on the seashore near high-tide mark. It thus ap-

pears that all of the hitherto supposedly different forms occur on the

seashore under similar conditions, and in addition certain fresh-

water habitats may be occupied, indicating a considerable range of

toleration for differences in the presence or absence of salt. In fact,

126 PAUL S. WELCH

Stephenson (11, p. 39) found that certain seashore forms from

Fintry Bay, which he recorded under the name L. subterraneus, “live

well for several days in a mixture of equal parts of salt and fresh

water, and equally well in altogether fresh water.”

Specimens of Lumbricillus lineatus were abundant, during July

and August, in a number of situations about Woods Hole, frequently

intermingling with specimens of Enchytreus albidus. In general,

the conditions of the habitats of the two species are much the same.

Protected coves with sandy, gravelly or rocky, sloping beaches, on

which the windrows of eel-grass accumulate near the upper tide limit,

provide favorable grounds for these worms. They seem to prefer

moderately moist conditions since the larger numbers occur near

high-water mark, and seldom at or below the low-tide limit. Their

usual position is just below the surface of the moist sand or gravel

and there is a distinct tendency to accumulate about the imbedded

stones or other larger objects to which they cling closely. Occasion-

ally, just after high tide, specimens may be found on the surface of

the sand below the overlying eel-grass which affords a protection

against sunlight and retains ample moisture to bridge over the low-

tide periods. However, specimens were found on semi-rocky

beaches where the eel-grass was quite thin or almost absent but the

worms were invariably deeper in the sand. The writer found this

worm very abundant in the protected coves in the vicinity of Hadley

Harbor. They also occurred in considerable numbers, intermingling

with E. albidus, on the beach just opposite the Lecture Hall of the

Marine Biological Laboratory. The two species can usually be

distinguished in the mature stage by the smaller size and particularly

by the reddish color of L. lineatus, E. albidus being invariably milk

white. However, it requires more careful scrutiny to distinguish

with certainty the immature forms since those of L. lineatus are

practically colorless.

METHODS OF CULTURE

These worms are readily kept alive in the laboratory for inde-

finite periods by employing the methods already described for E.

albidus.

THE ENCHYTRZIDZ OF THE WOODS HOLE REGION 127

MorPHOLOGY

Studies of the external and internal anatomy of these worms

corroborate, in most respects, the work of Moore. However, there

are some points of disagreement and, in addition, certain structures

and organs are described in detail for the first time. Therefore, only

those features of the morphology which demand special discussion

will receive consideration here and it will be understood that other-

wise the findings of the writer are in agreement with those of Moore.

Clitellum.—A well-defined clitellum is present in sexually ma-

ture specimens. Moore states that it “extends completely around

somites XI and XII.” This statement is evidently an error since

none of the numerous specimens examined by the writer has the clit-

ellum in that position but instead it occurs on XII and XIII. Its

completeness on the ventral side demands some additional descrip-

tion. Transverse sections through the penial bulbs or in their imme-

diate vicinity show the clitellum incomplete on the ventral side but

sections cephalad or caudad of the penial bulbs demonstrate that

this incompleteness is only partial and that the two ends of the clitel-

lum are continuous, although distinctly thinner on the ventral sur-

face.

Brain.—The description and figure given by Moore (05, p. 396;

fig. 24) present correctly the main features of this organ with the ex-

ception that only one pair of supporting strands are represented,

those extending from the posterior lobes. In reality, another pair

of slender supporting strands extends from the lateral margins, near

their middle, diagonally laterad to make attachment with the body-

wall. The posterior emargination, the angular form of the posterior

lobes bounding the emargination, and the very shallow concave form

of the anterior margin seem to be constant characters. The general

dimensions of the organ as a whole are subject to some variation, the

proportion of the length and width varying from 1.25:1 to 1.75:1.

Spermiducal Funnel.—The spermiducal funnel requires no addi-

tional description save in connection with the length. Moore (’05,

p. 397) states that this organ is about six to eight times as long as

thick. The writer has found considerable variation in the length,

depending upon the state of extension or contraction of the animal.

128 PAUL S. WELCH

In fully extended specimens, the above-mentioned dimensions are

approximately correct, but most of the specimens exhibit funnels the

length of which is only about four times the diameter.

Penial Bulb.—The external termination of the sperm duct is

described by Moore (’05, p. 397) as opening “into the small, de-

pressed, spheriodal, glandular and opaque atrium, which itself opens

on the medial side of a small bursa in the position of the ventral seta.

The bursa can be everted as a conical penis.” Properly translated

into the more modern terminology, the writer finds this very brief

description of the penial bulb apparatus to be correct as far as it

goes. The term “atrium” is applied to the organ which the writer

and others designate as the penial bulb, atrium being retained to

designate the enlargement of the sperm duct which appears in cer-

tain genera of enchytrzids just entad of the penial bulb. The

“bursa” evidently refers to the penial invagination.

Structurally, the penial bulb is simple and conforms to the lum-

bricillid type. Its transverse diameter is about one-fifth that of the

body in the same region. It is approximately globular and enclosed

in a thin musculature. The interior is filled with glandular cells of

one kind only. They differ somewhat in shape in the different parts

of the bulb but in general they are elongate, closely set together, and

most of them are arranged radially about the penial lumen. The

cells are distinctly nucleated, the nuclei being located for the most

part in the peripheral ends of the cells, although they appear in lim-

ited numbers nearer the penial lumen and well within the interior of

the bulb. The cytoplasm of the bulb cells takes artificial stains with

moderate intensity only. The sperm duct unites with the dorsal part

of the bulb near the body-wall and the former merges so gradually

into the latter that it is not possible to locate exact limits.

Ventral Glands.—These problematical organs, designated in the

literature under various names, such as ‘““Kopulationsdrisen’”, “copu-

latory glands’, ‘outgrowths of the ventral nerve cords”, “ventral

glands”, occur in this species but are subject to considerable varia-

tion in mature specimens. Well-developed glands occur in XIII and

XIV and sometimes in reduced form in XV. Most specimens con-

tain the first two but the one in XV appears only occasionally. The

shape and degree of development are also strikingly variable. Speci-

THE ENCHYTRAZIDA OF THE WOODS HOLE REGION 129

mens were examined in which the glands in XIII and XIV are pro-

duced into long wings extending dorso-laterad on either side of the

nerve cord, while the one in XV is diminutive and easily overlooked,

not extending above the level of the dorsal surface of the nerve cord

and developed mainly on one side only. Other specimens show only

the gland in XIV well developed and the one in XIII much smaller,

both being developed ventrad and laterad of the nerve cord but not

into wings, while no gland appears in XV. In all cases, the glands

are closely associated with the nerve cord, completely surrounding

it except for the median dorsal surface which is exposed. Moore

(705, p. 396) states that the “copulatory supra-neural glands are well

developed, especially in somites III, IV and V. They are slender

and elongated, not closely united with the ventral nerve, and open on

each side nearly at the ventral sete bundles.” The writer has found

nothing in III, IV, and V which can be identified as ventral glands,

the only thickenings of the nerve cord occurring in those somites

being due to the ganglia themselves.

Ovisac.—In the mature specimens in which there is an accumu-

lating mass of developing eggs, the septum XII/XIII is pushed

caudad to the posterior part of XIII. Whether this can justly be

designated as an ovisac may be open to question, but it probably rep-

resents the diminutive “ovisac” reported by Moore.

Spermathece.—The spermathecz are large, well-developed or-

gans lying in the usual position in V. Each is distinctly sigmoid in

shape and opens independently into the dorso-lateral aspect of the

digestive tract. There is no distinct demarcation between the duct

and the ampulla. The latter bears no diverticula, is fusiform in

shape, and contains a rather spacious lumen which corresponds to the

shape of the exterior. Both ectal and ental openings are distinct and

larger than usual. A complete crown of conspicuous glands sur-

rounds the ectal end of the duct and opens into the lumen of the lat-

ter at or just entad of the external surface of the body. This crown

surrounds practically the whole spermathecal duct, but is not directly

united with it except as above mentioned. The ental border of this

crown of glands is emarginate at more or less regular intervals, giv-

ing it a lobulate appearance and suggesting its origin from a number

of glands developed in very close proximity. This crown is made up

130 PAUL S. WELCH

of a large number of cells and each lobule is also multicellular. It

must be noted in this connection that Moore (’05, p. 397) found the

ectal end of each spermatheca surrounded by a “circle of glands

chiefly aggregated into an anterior and a posterior group.” His fig-.

ure of the same (Fig. 28) shows a very irregular crown of wnicellu-

lar glands, but the writer is inclined to suspect that imperfectly ma-

ture specimens might have been studied.

FRIDERICIA RATZELI (Eisen)

Identity

Worms, belonging to this species, were collected from the earth

and accumulating humus of the forest floor in Fay’s woods, back of

the village of Woods Hole. Difficulties arose in connection with the

identification of these enchytrzids, the chief obstacle being the far

too common one, namely, the vagueness and incompleteness of cer-

tain European descriptions. A complete morphological study.shows

that, with one minor exception, there is close agreement with those

characters of Fridericia ratzeli (Eisen) which have been described,

but, unfortunately, the anatomical discussions of the latter fail to

give any data concerning the following important features: origin of

the dorsal blood-vessel; position of the clitellum; extent and struc-

ture of the peptonephridia ; the chylus cells ; and the structure of the

penial bulb. In spite of these deficiencies, there seems no other

alternative, at present, than to consider the Woods Hole specimens

as belonging to ratzeli. The only point of disagreement is the origin

of the efferent duct of the nephridium near the posterior end of the

postseptal region, instead of near the septum as is described for the

foreign specimens of this species. The peptonephridia in the Woods

Hole specimens have fewer branches than seem to be called for in the

descriptions but it is very doubtful if any significance can be attached

to this variation. In addition, the posterior margin of the brain is

truncate rather than convex, but such variation is known to occur

in other species of Enchytrzidze. It seems, therefore, that the varia-

tions presented by the Woods Hole specimens are only of minor im-

portance and not of sufficient weight to demand a different identi-

fication. There is, of course, the possibility that when the foreign

THE ENCHYTR#IDZ OF THE WOODS HOLE REGION 131

representatives of F. ratzeli receive the much needed intensive mor-

phological study that a varietal difference between them and the spe-

cimens from Massachusetts may appear and, in order to facilitate

future comparisons, the morphology of the latter will be given in

some detail.

It should be noted that the specimens from Woods Hole corre-

spond very closely to an American species, F. californica, described

in detail by Eisen (’05, pp. 119-121) from specimens collected in the

vicinity of San Francisco, although certain points of disagreement

appear, such as the origin of the dorsal blood-vessel and the origin

of the nephridial duct. The chylus cells and the penial bulb are evi-

dently of the same structure in the California and Woods Hole speci-

mens. The writer is forced to the conclusion that californica is

identical with ratzeli, or if not identical, of no higher rank than that

of a variety.

This description records the second appearance of F. ratzeli in

North America, assuming that the writer is correct in placing

californica Eisen as a synonym of this species. The two localities

represented are separated by the width of the continent but this

might be expected since F. ratzeli is widely distributed, occurring in

England, Ireland, Scotland, Norway, Denmark, Germany, Switzer-

land, and Italy. The habits of this annelid appear to be much the

same in the different parts of its geographical range, being found

only in habitats of rich earth or damp moss.

EXTERNAL MorRPHOLOGY

The body is elongate, cylindrical, smooth, gradually decreasing

in diameter caudad of the clitellum. In nine sexually mature worms,

the length varies from 15 to 20 mm., the average being approximately

18mm. The diameter varies from 0.42 to 0.59 mm. Except in the

extreme anterior and posterior ends, the intersegmental grooves are

indistinct. The number of somites varies from 55 to 64, the aver-

age being 62. A well-developed clitellum occupies XII-XIII. The

sete bundles have the usual characters of the genus Fridericia. An-

terior to the clitellum, the ventral bundles contain 4-6 setz, usually

6, while the lateral bundles have a similar number, Caudad of the

132 PAUL S. WELCH

clitellum, the bundles contain 2-5 sete, the smaller groups occurring

near the posterior extremity. A head pore is present on 0/I and

dorsal pores begin on VII. The color of the living specimens was

very light yellow to milk white.

INTERNAL MORPHOLOGY

Brain.—This organ occupies a median, dorsal position in I and

II, chiefly in the latter. The anterior margin is distinctly convex,

the posterior margin truncate, and the lateral margins convex, the

maximum width being at a position about two-thirds the total length

‘from the anterior end. The length is about one and one-half times

the greatest width, typical measurements being as follows: length,

0.148 mm.; maximum width, 0.104 mm. Two pairs of supporting

strands extend from the organ to the body-wall, one from the lateral

margins near their middle, and the other from the caudo-lateral

angles. The description of F. ratzeli based on European specimens

calls for a convex posterior margin but since it is known that the

brain is subject to change of form under different conditions of con-

traction and extension, this variation can be of little significance.

Dorsal Blood-vessel—In the specimens examined, the exact

origin of the dorsal blood-vessel is a little difficult to determine. It

appears to become distinct from the perivisceral blood-sinus in XX,

or in the adjoining somites. The descriptions of ratzeli fail to in-

clude any statements as to the origin of this vessel in the European

specimens. Eisen (’05, p. 120) states that in californica it originates

in XVI.

Peptonephridia.—A pair of these organs occurs ventrad of the

digestive tract, one on the right side and one on the left. Each opens

separately into the alimentary canal in the extreme anterior part of

V and is confined to that somite. Sparse branching occurs at the

base and at the extremity of each. Aside from the branching, the

body of each gland is irregularly tuberculate. It is a matter of some

difficulty to determine just how closely these organs correspond to

the descriptions of foreign specimens of ratzeli. Michaelsen (00, p.

100) states that they are “mehrfach verzweigt”. Southern ('09, p.

164) finds them “freely branched” in specimens of ratzeli var. bed-

THE ENCHYTRZIDZ OF THE WOODS HOLE REGION 133

dardt from Ireland. Eisen (’05, p. 120) describes them in californica

as “narrow, slightly and irregularly branched”, although his figure

(Fig. 81, e) shows more branching than might be inferred from his

statement. The peptonephridia are known to be subject to a cer-

tain variation within a species and the relatively slight branching in

the Woods Hole specimens may merely represent one of the ex-

tremes.

Chylus Cells—The chylus cell region of the intestine occupies

XIV-XVI. Cells of two kinds are present, the chylus cells and the

ental epithelial cells. The former are somewhat flask-shaped, the

broader ends being ectad. The base of each is approximately trun-

cate and the sides converge gradually entad. The ental part of the

intracellular canal is usually straight or nearly so, but the basal por-

tion varies somewhat in its course, being usually bent to one side of

the cell and then curving abruptly, forming a right angle. The basal

part of the canal thus lies more or less parallel to the base of the cell

of which it isa part. This intracellular canal is lined by a relatively

thick specialized layer of cytoplasm which is everywhere of uniform

thickness and structure. The canal is ciliated for a part of its length.

A well-developed blood-sinus is present, surrounding the base of

each chylus cell and extending up the sides for about two-thirds of

its length. A conspicuous nucleus lies in the base of each chylus cell,

often a little to one side and in the angle formed by the chief bend

of the intracellular canal. The ental epithelial cells lie between the

apices of the chylus cells, the ental surfaces bounding a part of the

lumen of the intestine and bearing a thickly set coating of elongate

cilia. Interstitial cells are absent. The chylus cells are somewhat

longer in XIV than in XVI, making the whole layer thicker in the

anterior part of the chylus region.

A certain resemblance exists between the structure of the chylus

cell region in the Woods Hole specimens and in that described for

californica by Eisen. The exact position of the chylus cells, the

character of the intracellular duct, the general shape and character

of the ental epithelial cells, and the relative thickness of the layer are

the same, but lack of agreement exists in the California specimens in

the restriction of the blood-sinus to the bases of the cells, the close

apposition of the cells, and the approximately parallel sides. Chylus

134 PAUL S. WELCH

cells do not seem to have been described for the European specimens

and comparison is not possible.

Nephridia.—The anteseptal and postseptal parts are about the

same size although some slight variation occurs in the different —

somites. The efferent duct arises from the mid-ventral surface of

the postseptal part, near the posterior end. Some variation also

exists as to the origin of this duct but it was not found farther for-

ward than the middle of the postseptal region. The internal struc-

ture is very similar to the usual type found in the genus Fridericta.

Foreign specimens of ratzeli are described as having the origin

of the nephridial duct on the anterior portion of the postseptal part

near the septum. Eisen’s description of the nephridia in californica

contains no statement as to the exact origin of the duct but his figure

(Fig. 81, a) shows it as arising near the middle of the postseptal

region.

Spermiducal Funnel.—A pair of these organs lies in the usual

position in XI with the bases close to the ventral part of X/XI and

the free extremities directed cephalad. The length of each is about

two to three times the maximum diameter. A small, reflected collar

is distinctly set off from the body of the funnel by a constriction, its

diameter being only about one-half that of the body of the funnel.

Each sperm duct is confined to XII and presents a mass of convolu-

tions in the ventral part of the ccelom.

Penial Bulb.—Structurally, the penial bulb is of the lumbricillid

type. It is small, compact, globular, and completely invested by a

thin but definite musculature which is derived from the muscle-layer

of the body-wall. When completely retracted, the invagination is

longer than the transverse dimension of the organ, the latter lying

completely on the mesal side. The sperm duct unites with the penial

bulb on the dorso-ectal surface, penetrating it for a short distance.

Within the bulb, it is replaced by the penial lumen, which, curving

abruptly laterad, extends to the penial invagination, opening into its

ental part. The bulb is composed of cells of but one kind, these be-

ing large compared with the size of the organ and distinctly nu-

cleated.

The structure of the penial bulb is similar to that described by

Eisen (’05, p. 120) for californica in being composed of cells of only

THE ENCHYTR#ZIDA OF THE WOODS HOLE REGION 135

one kind, but in the retracted condition, the sperm duct cannot be

said to “enter the bulb near the base, splitting the bulb into two un-

equal parts.” Furthermore, if Eisen’s figure (Pl. XV, Fig. 9) was

made from a transverse section of the worm, a distinct difference

appears in the shape of the invagination and the organ as a whole.

Mention of the bulb (‘prostate’) in the descriptions of the Euro-

pean specimens is so brief that little can be done in the way of com-

parison. It is said to be almost as large as the spermiducal funnel, a

proportion which in the Woods Hole specimens would make the bulb

several times its present size.

Spermathece—A pair of well-developed spermathece is pres-

ent in V, each organ being composed of three regions: duct, diver-

ticula, and ampulla. The ectal opening of the duct is in the usual

position in the lateral aspect of IV/V and has associated with it two

small, multicellular, pyriform glands. The ampulla bears on its ectal

end a circle of irregular, sessile diverticula, usually eight in number,

which vary somewhat in shape but are mostly globular and pendant.

Excluding the expanded portion bearing the diverticula, the ampulla

_is distinctly spindle-shaped. Each ampulla opens independently into

the dorso-lateral aspect of the digestive tract. In the sexually ma-

ture specimens examined, the ampulle contained spermatozoa and,

in some cases, spermatozoa were not only in the process of passing

into the lumen of the alimentary canal, but masses of them were

found well within it.

Ventral Glands——In XIII, XIV, and XV, certain problematical

glands occur in connection with the ventral nerve cord. They com-

pletely surround the ventral and lateral parts of the latter, leaving

only the dorsal surface exposed. At first sight, they appear as

unusually developed ganglia, but stained, transverse sections show

the tissues of the nerve cord to be distinctly different from those

which immediately surround it. These glands are made up of large,

nucleated cells, massed irregularly together, which give off processes

extending ventrad through the body-wall to the hypodermis. The

latter assumes a finely striated appearance with hypodermal nuclei

appearing only at irregular intervals. These gland cells are many

times larger than the nerve cells immediately above them. The

136 PAUL S. WELCH

organs are not of uniform size, the one in XIII being larger than the

subsequent ones.

Similar glands, described under the various names mentioned on

a previous page, occur in certain other genera of enchytreids, namely |

Lumbricillus and Marionina (Welch, ’14, pp. 150-151), but their ap-

pearance in Fridericia is quite unusual, being known only in two or

three species. Ude (’01, pp. 6-7) found one ventral gland in XIII

in the specimens of ratzeli which he examined. They have been re-

ported in XIII-XV in F. antarctica Bretscher, a synonym, according

to Michaelsen (’00, p. 100), of ratzelt. Fridericia dura (Eisen), an-

other synonym of ratzeli, according to Michaelsen (’00, p. 100), but

a distinct species according to Ude (’01, pp. 6-7) possesses one ven-

tral gland in XIII.

It should be noted in passing that, in the Woods Hole specimens,

superficially similar swellings of the ventral nerve cord occur in II,

III, and IV. However, a study of sections through these anterior

swellings shows that they are nothing more than ganglionic assemb-

lages of nerve cells and not special masses of gland cells.

SuMMARY

1. The enchytrzid fauna of the Woods Hole region, as known

at present, comprises three species, Enchytreus albidus, Lumbricillus

lineatus, and Fridericia ratzeli. The first two are marine and occur

in great abundance in the inter-tides zone, especially in the protected

coves which accumulate windrows of eel-grass. The other species is

a terrestrial form, inhabiting the soil and humus of forest land.

2. Both of the marine species can readily be cultured indefin-

itely in the laboratory by placing them in shallow stender dishes con-

taining a small amount of sea water and pieces of green eel-grass

from the drift on shore. They could not be kept alive in the fresh

water taken from the Marine Biological Laboratory supply, although

when a few drops of sea water were added the unfavorable condi-

tions were relieved.

3. Detailed study of the morphology of the sexually mature

Lumbricillus yielded evidence that the foreign forms which have

been discussed under the names lineatus, subterraneus, verrucosus,

THE ENCHYTRZIDZ OF THE WOODS HOLE REGION 137

and litoreus are all one species, and that the American species, agilis,

originally reported from Woods Hole also has the same fate. These

should all be regarded as one species under the name lineatus which

has priority.

4. Fridericia ratzeli has evidently been discovered in North

America before, under the name of californica, which was applied to

specimens found near San Francisco by Eisen. Critical morpholog-

ical study of the Woods Hole specimens makes it almost certain that

californica is a synonym of ratzeli.

5. All of the three enchytrzids herein mentioned are widely

distributed in the Old World and evidently so in North America.

6. Morphological detail not readily summarizable are presented

for two of the species. Special consideration has been given to the

organs of the complex reproductive system, the chylus cells of the

digestive tract, and the ventral glands.

BIBLIOGRAPHY

Derguat, L.

1914. Gli Enchitreidi della Toscana. Estratto dal Montore Zoologico

Italiano, 25 :13-24.

DiTLEvseNn, A.

1904. Studien an Oligochaten. Zeit. f. wiss. Zool., 77 :398-480. 3 pl.

Ersen, G.

1905. Enchytrzide of the West Coast of North America. Harriman

Alaska Expedition, 12:1-166. 20 pl. New York.

Frienp, H.

1912, New British Oligochets. The Zoologist, (4) 16 :220-226.

1916. Notes on Irish Oligochets. Irish Naturalist, 25 :22-27.

Hunt, H. R. |

1915. Regeneration Posteriorly in Enchytreus albidus. Am. Nat.,

49 :495-503.

MICHAELSEN, W.

1900. Oligocheta. Das Tierreich, 10 Lief. XXIX+575 pp. 13 fig.

Berlin.

138 PAUL S. WELCH

1901a. Hamburgische Elb-Untersuchung. Zoologische Ergebnisse der

seit dem Jahre 1899 vom Naturhistorischen Museum unternom-

menen Biologischen Erforschung der Niederelbe. IV. Oli-

gocheten. Mitt. Nat. Mus. Hamburg, 19:169-210. 1 pl.

1901b. Oligocheten der Zoologischen Museen zu St. Petersburg und

Kiew. Bull. Acad. Imp. Sci. St. Petersbourg, (5) 15 :137-215.

1909. Oligochzta. Die Siisswasserfauna Deutschlands, Heft 13:1-66.

112 fig.

1914. Beitrage zur Kenntnis der Land— und Siisswasserfauna Deutsch-

siidwestafrikas. Ergebnisse der Hamburger deutsch-stidwest-

afrikanischen Studiesreise 1911. Oligocheta, 139-182. 1 pl.

Moors, J. P.

1905. Some Marine Oligocheta of New England. Proc. Acad. Nat. Sci.

Phil., pp. 373-399. 2 pl.

SmitTH, F.

1895. Notes on Species of North American Oligocheta. Bull. Ill. State

Lab. Nat. Hist., 4 :285-297.

SouTHERN, R.

1909. Contributions towards a Monograph of the British and Irish

Oligocheta. Proc. Royal Irish Acad., 27 :119-182. 5 pl.

STEPHENSON, J.

1911. On some Littoral Oligocheta of the Clyde. Trans. Royal Soc.

Edinburgh, 48:31-65. 2 pl.

Sumner, F. B., Ossurn, R. C., and Coxe, L. J.

1913. A Catalogue of the Marine Fauna. Section III, Part II. A Bio-

logical Survey of the Waters of Woods Hole and Vicinity.

Bull. Bureau of Fisheries, 31 :547-860.

Upr, H.

1901. Die arktischen Enchytraiden und Lumbriciden sowie die geo-

graphische Verbreitung dieser Familien. Fauna Arctica, 2:1-

S42 ph tin

VerriLL, A. E.

1873. Report upon the Invertebrate Animals of Vineyard Sound and the

Adjacent Waters, with an Account of the Physical Characters

of the Region. Rep. U. S. Fish Comm., 1871-72, pp. 295-747.

38 pl.

WE cu, P. S.

1914. Studies on the Enchytreide of North America. Bull. Ill. State

Lab. Nat. Hist., 10:123-212. 5 pl.

CHEMICAL MICROSCOPY*

By Dr. E. M. CHAmor

Department of Chemistry, Cornell University, Ithaca, N. Y.

A speaker who has the temerity to address a joint meeting of

two different technical societies always finds himself in an awkward

predicament. He feels that he must present his subject from the

viewpoint of each group of men, that he must lay equal emphasis

upon all branches of the sciences represented by his audience. I find

myself very much embarrassed, realizing this, and in doubt whether

I am here in the guise of a microscopist or of a chemist.

This, gentlemen, is the introductory paragraph of the paper I

had originally prepared. Since that time we have entered the great

war and I know where I stand. I come to you as a chemist to make

an appeal for a wider and more intelligent application of the micro-

scope in every day chemical practice.

If the talk appears rambling and fragmentary I trust you will

bear with me, for with several momentous issues on the hands of my

department I have had little opportunity to prepare a new paper and

none to make new lantern slides. I will, however, attempt to stick

to my text—Chemical Microscopy. To my mind there is no such

thing as microchemistry as opposed to macrochemistry and the term

microchemical methods is a misnomer. A microchemical reaction or

test may be one performed upon minute amounts of material with-

out necessarily having recourse to the microscope.

Chemical Microscopy on the other hand requires that some type

of magnifying optical instrument enters into the work. By chemical

microscopy, therefore, we simply mean the application of micro-

scopic methods to the solutions of problems arising in the chemical

laboratory or in the chemical industries.

No instrument at our command can do so much or throw so

much light upon obscure problems with so little expenditure of time,

labor and material. We chemists have been wasting golden hours

and slaving over sloppy methods to accomplish ends which could

*Address delivered before a joint meeting of the Chicago Section of the American

Chemical Society, and the State Microscopical Society of Illinois, at the City Club, Chicago,

on April 20th, 1917; Messrs A. V. H. Morey, Chairman of "the Chemical Society and

N. S. Amstutz, President of the Microscopic Society, presiding. A number of self explan-

atory plates accompanying this article are found as inserts following page 156.

140 E. M. CHAMOT

have been reached easily and leisurely and with a degree of certainty

unsurpassed by anything we have had at our command with test

tubes and beakers. Why spend hours upon a qualitative analysis

that can be better done through the medium of the microscope in

several minutes?

The time has come when we can no longer be satisfied with time-

consuming operations. Industries must be speeded up, production

increased, better inspection methods introduced, quality thereby

raised and final cost in consequence reduced. Iam simply preaching

good conservation. If we fail in this war and disappear as a nation

it will be because we have failed in our industries to produce the

necessary material and the requisite quality. I cannot recall a single

great industry today where microscopic methods intelligently applied

will not lead to more or less marked improvements.

The causes of our failure fully to appreciate the value of micro-

scopic methods are not hard to find. In the first place in the educa-

tional system of our chemists no adequate training has been given

in the multiplicity of uses of the microscope and its potential indus-

trial applications. In the second place too much emphasis has been

laid upon biologic microscopy; so that the generally accepted view

is, that this instrument is intended for studies in biology or medicine.

As a result, the development of the modern so-called high grade

microscope has followed strictly biological lines and has drifted far-

ther and farther away from stands applicable for general work in the

chemical laboratory. For example the best of our present day

stands no longer have the mirror mounted upon a swinging bar capa-

ble of movements far to one side, or even above the stage for oblique

illumination. In refractive index work, in the observation of melt-

ing points, in the study of fatigue failure in metals, in the general

examination of alloys, cements, protective coatings, etc., and in the

preparation of photomicrographs of certain preparations, this old de-

vice now abandoned is really essential, and it is necessary to remove

the mirror from the stand and fasten it to a holder of some sort that

it may be properly employed.

It is also unfortunate that the objective of small angle, long

available working distance and marked penetrating power are not

obtainable save at second hand. The modern microscope objective

CHEMICAL MICROSCOPY 141

is a marvel in its performance, yet it is limited to the study of

mounted materials covered wtih a standard cover glass if over a mod-

erate power is required for the examination; but unfortunately, we

chemists must work with uncovered preparations and we must sac-

tifice resolving power for penetrating power and for stereoscopic ef-

fects. We need instruments of moderate cost, substantially built,

and which will withstand the corrosive atmosphere of most of our

industrial laboratories. Thus the third reason for the backward-

ness of chemists to use the microscope has been the lack of suitable

models and accessories. Even our good friends and near chem-

ists, the petrographers, have never gone out of their narrow way,

to try to impress the chemist with the fact that the polarizing micro-

scope is an indispensable adjunct of the research laboratory. The

modern petrographic microscope is a measuring instrument of great

precision. By its means alone a vast number of chemical compounds

can be positively identified. The manufacturer of organic com-

pounds, especially, cannot afford-to ignore it as a means of increasing

the ease of control work. In the hands of a skilled worker this type

of instrument offers untold advantages.

The application of microscopic methods to analytical problems

should appeal to every chemist. Not only can he perform qualitative

chemical analyses easier, but he can measure refractive indices of

both solids and liquids, determine melting and boiling points with

exceptional accuracy and upon minute amounts of material which

cannot be isolated; determine molecular weights and can study the

structure of most of our commercial materials. Dr. Harvey W.

Wiley, in one of his happy moods, once defined chemistry as “the

astronomy of things infinitely small.” Our telescope is the micro-

scope. To make suitable progress we must, like the astronomer,

construct special instruments for special purposes and like the

astronomers we must become specialists in narrow fields within our

vast science. We chemists must have analogues to the students of

double stars, to the investigators of nebulz, to the seekers for comets,

etc., when this day comes the results reported will be comparable

to the discoveries of our astronomical friends. Americans make the

finest telescopes in the world. Why not microscopes as well? Un-

fortunately the scientific world in the United States has been

142 E. M. CHAMOT

obsessed with the idea that no microscopes are worth using, unless

made in Europe. We are all to blame for the present difficulties.

Can you obtain an ultramicroscope, a luminescense microscope or

even a polarization microscope in the United States this 20th day of —

April? Not one, nor can you obtain condensers, lenses and eyepieces

of quartz suitable for photography with ultra violet rays, nor spec-

troscopic oculars ; nor can be purchase a really satisfactory moderate-

priced metallograph, although in this line there is hope that instru-

ments will soon be on the market.

If each one of us here tonight will agree hereafter to stand by

American manufacturers and buy American made instruments, we

will soon have special microscopes and accessories ranking with the

best obtainable. Our artisans have no superiors and few equals, but

in order that we may persuade them to undertake the construction

of the apparatus we require it is essential that we must support them

with advice and hard dollars and not with empty applause. It is

easy to find fault and refuse to cooperate; but it takes time and tact

to call attention to defects and suggest improvements.

If those using special microscopes would stop and consider the

care and labor involved in their construction and would be a trifle

more tolerant toward mistakes in construction, far greater progress

would be made than at present. Let us all agree to try and stimulate

the development of American types of microscope which will do our

work better and easier, and cease being mere copyists. Let us be-

come “boosters” instead of “knockers”.

I have already asserted this evening that the chemical micro-

scope will do more for the chemist than any other instrument or

group of instruments, and it behooves me to prove my contention.

In the first place microscopic methods are the simplest and shortest

for the identification of a compound. Let us assume that the analyst

has in his hands a crystalline salt, and by qualitative analysis in the

usual manner he decides after about an hour’s examination that it

contains sodium and phospheric acid—nothing else. It is manifestly

a sodium phosphate, but which one? Mono, di or tri? This he can

answer satisfactorily by quantitative analysis, and actually only by

a determination of Na and PO,. If, however, he possesses a polar-

izing microscope, the problem is quite simple. The mono-sodium salt

CHEMICAL MICROSCOPY 143

is orthorhombic, the di-sodium, mono-clinic, while the tri-sodium

phosphate is hexagonal. He can clinch his opinion with one or more

simple optical measurements and prove his case by refractive index

determinations by the immersion method. Why is it that the chemist

never uses refractive index determinations by means of the micro-

scope as an aid in qualitative analysis? It is inconceivable that we

have had these methods used for years by mineralogists and petro-

graphers, yet never had sense enough to apply them to our own ends

and thereby save ourselves hours of time.

But to go back to our phosphate; had we made the qualitative

analysis by microscopic means it would not have taken us an hour,

but say not over half that time. We would have been seated com-

fortably at a table and would have satisfied ourselves in a very few

minutes that the salt was di-sodium phosphate badly effloresced and

of commercial, not C. P., quality. This case is actually a typical one

and very simple. I have selected it because it illustrates quite clearly

the way in which a simple salt may be identified. But I hear some of

you say this requires a knowledge of crystallography. What of it?

If this knowledge will save us time and labor let us by all means do

a little reading.

Of all the inorganic salts we will meet with in industrial work

only a very few belong to the isometric system and have no effect

upon polarized light. Very few tetragonal and triclinic and fewer

still hexagonal. There is rarely need for expert training to enable

the analyst to properly place the compound under examination in

one of these systems. Suppose again the analyst has an inorganic

crystalline salt, and under the microscope it separates from water

in what appears to be large colorless octahedra which are isotropic.

The salt must be an alum, or strontium, barium, or lead nitrate, or

one of several chlorostannates. The addition of a tiny drop of

nitron sulphate gives no crystalline precipitate, therefore it cannot

be Sr, Ba or Pb, or other nitrate. A little calcium acetate gives crys-

tals of calcium sulphate. The salt is presumably, therefore, an alum.

A refractive index determination will show which alum, or we can

go ahead and test qualitatively for the bases present.

The point I wish to emphasize is that in the identification of

many substances a systematic time-consuming analysis is unneces-

144 E. M. CHAMOT

sary. Note well also that all the work is done upon an object slide,

that only low powers are employed and the amount of reagent re-

quired is negligible. Five grams of practically any reagent used

should last an analyst, even in daily examination, almost a lifetime.

I find that the prevailing idea among chemists is that qualitative

analysis, by means of the microscope, has for its purpose, the detec-

tion of infinitesimal traces of material such that all other methods

fail. Although it is true that microscopic methods can be thus

employed, by far the greatest points in their favor are the rapidity

of obtaining results and the certainty of reaction. Actually the rela-

tive proportion of material to solvent is very great, we are apt to

be working with high concentrations. We take a fragment of the

unknown material, not quite as large as a pin-head, and dissolve it

in a minute drop of water or acid. The microscope enlarges this

drop of solution until it appears to have the diameter of a twenty-

five cent piece. This is almost equivalent to taking a handful of

the unknown and dissolving it in a litre of solvent. The identity

test is made by adding a reagent which will lead to the formation

and separation of a crystalline phase.

It is in the field of organic analysis that the microscope stands

without any possible competitor. Differentiation of isomeric com-

pounds, recognition of different degrees of sulphonation, nitration,

etc., is so simple in most cases as to be mere child’s play. Take the

case of the phenol sulphonic acids. Recognition of the different

acids, mono or di, ortho, meta or para or mixtures, was a stumbling

block for years until Pratt showed that the barium salts were easily

differentiated under the microscope. The di-acid salt forms stout

mono-clinic prisms, the mono-ortho acid long slender rods, and the

para acid tufts of fine needles. Quite recently the microscope was

called upon to aid a large plant in controlling the completion of a

certain process. It was found that the manganese salt of a certain

organic compound crystallized in plates with vivid polarization col-

ors; the other product which it was desirable to eliminate crys-

tallized only with difficulty in sphzro crystals polarizing feebly. A

glance under the polarizing microscope showed at once the stage in

the transformation of one form into the other. The older method

CHEMICAL MICROSCOPY 145

of control took not less than twenty-four hours; the new not over

twenty minutes.

By far the majority of organic compounds cannot be differen-

tiated and identified without time consuming quantitative determina-

tions. Judicious application of the methods of what Lehmann years

ago called crystal analysis yields the necessary information at once.

By means of an electrically-heated stage not only can we deter-

mine melting points with greater ease than by the usual methods,

but the accuracy of our observations is considerably increased.

Amounts of material so small as to be practically invisible to the

naked eye can be employed. A further advantage lies in the fact

that the melting points of several different substances existing in a

mixture may be ascertained without having recourse to long and

arduous separations, involving loss of material and time.

Very reliable melting points of fats may also be obtained as well

as the boiling and subliming points of small quantities of material.

The advantages of microscopic melting point determinations

will become more apparent when we recall that when we separate

one compound from another at our laboratory work table, we so

proceed that the final products stand upon our table a few inches

apart in suitable containers. If we spread a small quantity of the

original mixture upon a bit of cover glass and examine the prep-

aration with a magnification of say fifty diameters, a decided space

will be seen to exist between most of the different components.

Gentle tapping will usually increase this space. To all intents and

purposes the magnification has done exactly what we accomplished

in our long chemical separation ; i. e., removed the components from

apparent contact with one another, and interposed space between

them. In most instances even very rapid crystallization of two or

more salts upon a slide by quick evaporation of their solution will

yield a preparation in which the salts will be found to have separated

without intermixture, and with a sufficient space between them to

allow of a melting point determination being made.

This fact is clearly shown in Fig. 1. Evaporation has been

pushed so fast as to force the salts to crystallize in dendritic forms,

yet each group of dendrites is clear and distinct. Were such a

preparation heated carefully and watched with the microscope, I

146 E. M. CHAMOT

think you will all agree that as one of the components begins to

melt it will easily be discerned and will not interfere with the other.

When, however, the temperature is raised to the melting point of |

the second component, the chances are that the two liquids will

flow together. Nevertheless the moment of fusion is easily ascer-

tained. If in melting point observations, as I pointed out some

years ago, we make use of the polarization microscope, the transition

from a solid anisotropic body to a completely fused isotropic body is

instantly recognized.

Doubtless that branch of chemical microscopy of greatest gen-

eral applicability is in qualitative analysis. The addition of a suit-

able reagent induces the formation of a distinctive compound and its

subsequent separation as a solid crystalline phase. These crystals

are easily recognized and are so characteristic that there is little

danger of mistaking those given by one element or compound for

those of another. Add to the distinctive morphology the fact that

color also enters into the identification scheme, and it will be even

more apparent why microscopic methods offer such ready means of

identification. In a large number of cases one and the same reagent

will cause. distinctive crystals separations with a number of sub-

stances. One of the best examples of this is potassium (or sodium)

mercuric sulphocyanate, K, Hg(CNS), (or 2 KCNS. Hg (CNS)2)

which gives characteristic crystals with copper, yellowish green

(Fig. 2) ; cobalt, deep blue, (Fig. 3) ; zinc, white (Fig. 4) ; cadmium,

colorless (Fig. 5); lead, colorless (Fig. 6); manganese, colorless

(Fig. 7); gold, yellow; silver tiny colorless; and a red color with

iron. Thus the addition of a single reagent will show at once the

presence or absence of a number of elements, and at the same time

produce an identity test for each, thereby saving an enormous

amount of time and material. There are a number of such reagents

available, and by carefully choosing them we can complete in a

few minutes a qualitative analysis, intricate though it may be.

Time will not permit me to show slides of more than one or more

of these multiple test reagents. I have selected cesium chloride,

which gives us reactions for bismuth, antimony, tin, copper, silver

and lead, and occasionally, aluminum and magnesium. You will

note that the crystals obtained are just as different from each other

CHEMICAL MICROSCOPY 147

and just as easily recognized as those formed by the mercuric sulpho-

cyanates. The slides I now show you are the characteristic reactions

for the common acids. You will note that the crystal forms are so

different and so easily remembered that there can result no confusion

when the tests are properly applied.

The amount of material required for our tests is shown by the

tiny fragment clinging to an ordinary No. 7 sewing needle (equiva-

lent to a fragment whose diameter is approximately that of a period

(.) on this page). The manner in which a test is performed I have

tried to show in this lantern slide, Fig. 8. An ordinary 3x1” object

slide with the steps in the analysis of an alloy has been photographed

natural size in order that the relative sizes of the drops may be bet-

ter judged. The tiny black spot (0.8 mm. x 0.1 x 0.1 mm.) is a piece

of the alloy of the exact size of that employed for the analysis. The

large spot at the corner is the space occupied by the solution of the

alloy after repeated evaporations with tiny drops of nitric acid to

render insoluble any tin present. The nitric acid soluble portion

has been decanted to the second spot. The residue has been tested

and found to contain tin, antimony, and copper. The nitric acid

solution has been divided into three drops as seen on the slide; the

first shows the dry residue after finding lead, copper and antimony

present; the third spot is what remains on the slide after testing

for other elements and finding zinc and iron in traces, in addition to

those already found. The fourth spot has been used for testing for

the remaining possible metals which were not disclosed in the other

tests.

We have thus carried out upon an object slide the entire quali-

tative analysis of a bearing metal containing tin, antimony, lead, and

a small amount of copper, and having traces of iron and zinc present.

Actually it took no more time to perform the analysis than it has

taken time to tell it. The worker has been seated at a small table

and has used less than five cents worth of reagents and gas. Instead

of cutting off a small portion of the alloy we could just as well

have rubbed it over a piece of ground glass or unglazed porcelain,

dissolved off portions of the streak with acid and made our analysis

as just outlined. The next lantern slide shows this method together

with the results obtained and the time required for the analysis.

148 E. M. CHAMOT

These cases are fair illustrations of what is possible in the saving

of time, money and labor, through the employment of microscopic

methods.

Other valuable applications of qualitative tests are those involv-

ing testing for the purity of precipitates in gravimetric analyses in

order to avoid the time and trouble involved in resolution and re-

precipitation, in testing for complete precipitation, especially in elec-

trolytic analysis and also in testing for complete washing.

Another valuable application of microscopic chemical methods

is in the analysis of the total solid residue in water analyses. We

generally speak of the hypothetical combinations present. I do not

wish to raise the question of reporting ions or combinations, but I

do desire to lay emphasis upon the fact that it is possible and practi-

cable, in most cases, to identify the salts present in the solid residue

through their habit and optical properties, providing the work is

properly done. I know of several instances where identification of

the principal compounds present threw much light upon obscure

problems. Traces or more of the heavy metals, such as lead, cop-

per, etc., are far more readily detected by microscopic qualitative

analysis methods than by any other means at our disposal.

Water analysts pay altogether too little attention to microscopic

examinations of sediments and suspended matters, to the deposits

at the bottom of springs, wells and cisterns. Much valuable informa-

tion is also to be derived from the examination of the muddy ooze

at the bottom of streams, ponds and reservoirs, and from the study

of the coated sands from rapid filter beds, to learn the extent and

character of the adsorption of aluminum hydroxide by the sand

grains. When we speak of the “microscopy of drinking water”,

we generally mean researches upon the flora and fauna giving rise

to disagreeable odors and tastes, but this is in reality only a very

narrow portion of a huge field which is by no means restricted to

biological problems or even to investigations made with the ordi-

nary microscope, since it comprises problems soluble only by means

of such specialized instruments as the ultramicroscope and the lum-

inescence microscope. |

The remaining lantern slides have been chosen to illustrate the

application of microscopic methods to the solution of problems in

CHEMICAL MICROSCOPY 149

some of the great industries. I can do no more than touch upon

them. Permit me, therefore, to merely outline the nature of the

information given.

Abrasives: Proper grinding requires adequate speed without

undue heating, cutting of uniform depth, wheels which wear well.

In other words the selection of the proper sort of wheel and speed

for the specific purpose. There enters in addition, the size of par-

ticles of abrasive and the nature of the bonding material giving a

hard or soft wheel. Much of the manufacturing has been done upon

a purely empirical basis and by rule of thumb methods.

Microscopic examinations of particles torn off shows how the

wheels have acted. While a similar examination of the ground sur-

face shows the character of the cutting done.

It is surprising how much information may be gained in this

way, and how it may be used to guide one in making proper

selections. | |

I will be able to demonstrate that a grinding wheel of a certain

kind will tear off the surface of tool steel in such a manner as to

heat the steel and draw its temper to such a degree that the particles

you will see under the microscope have been fused into tiny spheres.

Another wheel rotating with the same surface velocity will cut off

the material in ribbons. You will note how few melted fragments

are present, as shown by the absence of tiny spherical masses. Such

a wheel can be employed for purposes for which the other is obvi-

ously unsuited.

Cement, Concrete, Ceramics, etc.—By microscopic examination

it is possible to determine the character of the final product, its com-

ponent parts and their volume per cent; the prevalence of an undue

proportion of air and water voids; the thoroughness of wetting of

the cement mixtures, etc.

Not less important is the recognition of improper bonding and

valuable information is obtainable as to the actual strength of the

concrete or its liability to failure. There is here a huge field for

the investigator offering untold possibilities.

The whole field of ceramics, both clay products and porcelains,

needs intensive microscopic research. Even our ordinary bricks

offer a most attractive subject for the investigator.

150 E. M. CHAMOT

The thickness of glaze and the thoroughness of its bond with

the body-making material can readily be determined. I have pre-

pared slides of two high-grade porcelains; in one of these, Fig. 9,

you will note the glaze is thick and between it and the porcelain

are a vast number of tiny gas voids. In some cases these gas voids

penetrate the glaze as infinitely fine capillary tubes. The other

porcelain of far higher quality has a much thinner glaze and as you

see, (Fig. 10), has almost no gas voids, the bonding is almost per-

fect. Examinations of this sort, employed in the industry, go a

long ways to improve the products turned out.

Foods and Beverages: Doubtless the earliest application of

microscopic methods by the chemist was in the examination of

foods, food accessories and drugs for adulteration or deterioration.

Examinations of this sort are based largely upon vegetable and

animal histology, and differ but little, if any, from the ordinary

technique of histology. No other methods are available to accom-

plish the ends in view, and we can therefore assume that in this

line at least the microscope is indispensable, training in the neces-

sary technique imperative, and every food analyst and expert must

perforce train himself to undertake studies of this sort.

This field of activity is so well understood and so firmly estab-

lished that we can dismiss it without further comment. But there

is in this whole question of the microscopy of foods and drugs,

another phase, which chemists have greatly neglected, that of the

amazing possibilities of microscopic qualitative and quantitative

analysis. To even enumerate the list of substances whose detection

becomes simpler, more certain and much more rapid than by our

ordinary routine tests, would require more than the remaining time

at my disposal. These “micro” tests are applicable to organic, as

well as inorganic substances, as I have already pointed out. We

have nothing better than microscopic methods for the detection of

the poisonous metals, for the recognition of the organic acids, for

the detection of preservatives, for the vegetable alkaloids, glucosides,

and other active principles of plants; nor can we find other methods

available for the quantitative analysis of starch mixtures and for

similar analyses of mixed powders and meals of vegetable origin.

CHEMICAL MICROSCOPY 151

The recognition of our commercial synthetic drugs also is con-

siderably simplified. In fact it is no exaggeration to say that proper

analyses of this sort cannot be conducted unless the microscope is

employed. .

In the canning industry, especially that employing tin cans and

other containers, the microscope gives information of untold value.

Soldered and crimped joints yield up their secrets, as also the tinned

surface or other protective coating which may have been applied

to metal or paper surfaces. The possibilities, are in fact, without

end.

Metallurgical Industries: The microscopic study of metals and

alloys has been so firmly established within the last few years, and

the close relation between structure and physical properties so gen-

erally recognized and its importance proved in practice, that I need

not spend time upon this question. Although these methods have

been placed upon a firm foundation in the iron and steel industries,

there is much work to be done in the great field of commercial alloys.

We generally think of heat treatments in terms of steel only. As

a matter of fact many alloys may be greatly improved by carefully

conducted heat treatments. In order that such work may be properly

done, microscopic studies of structure are imperative. This is well

shown in photographs of a copper, zinc, and of a copper-aluminum

alloy, which I will show you.

Often the microscopic appearance of a roughly polished and

etched specimen taken in conjunction with a hasty qualitative analysis

will give the investigator all the information he may require to de-

duce the quantitative composition, and to make a shrewd guess at

the physical properties. |

A most fruitful field leading to improved practices is the study

of welds and brazes under the microscope. At least one expert in

welding by means of oxy-acetylene owes much of his remarkable

success to microscopic studies of welded materials. |

Not infrequently the microscope shows that a poor product is

due to improper temperatures of casting or coating, and not to bad

raw materials or wrong percentage composition. This is especially

the case in babbitts and in tinned goods. We have here photographs

152 E. M. CHAMOT

showing the great difference in the structure of a babbitt cast at too

high a temperature and the same one poured just right. It is obvious

that in the one case friction will be considerably greater than in the

other, particularly if in a high speed bearing. i,

Paints, Pigments, Protective Coatings: The microscopic studies

of materials falling in this class may be classed under three heads:

(1) The examination of the raw materials, (2) that of the coated

surfaces and (3) studies of the methods and results of applying the

coatings to the objects to be protected.

Briefly stated the raw materials under the microscope (chiefly

pigments, etc.,) reveal their source and nature; often the process

of manufacture and their suitability for the purposes for which they

have been purchased. Take for example the mere question of size

in the selection of the pigments for a mixed color paint. It is a sim-

ple matter to obtain a whole series of different shades by using, say

two pigments and having them vary in the ultimate size of their

particles, although the per cent by weight of each remains constant.

The actual shape of the particles also probably seriously effects

the length of time the paint really acts as an efficient protector,

especially in paints containing silica, graphite or both. The micro-

scope also throws light upon the nature of the changes taking place

in the pigments of paints on being exposed to air, light and the

weather. A good illustration is to be found in the study of the cause

of the darkening of lithopones.

Examination of the weathered, coated, objects, both of the

surfaces and of section, cut through the thin films of coatings, not

infrequently will permit the formation of an immediate opinion as

to the quality of the paints or coatings, and the skill of the workmen

who applied them.

Too little attention has been paid in the past to the study of

sections. 1 believe you will all be interested in the prepara-

tions I have to show you. I have selected them because they exhibit

in a striking manner the differences between good and bad paints,

and between good and bad workmanship in their application. You

‘will also note that the appearance under the microscope of these

samples of japanned steel show very clearly the superiority of the

CHEMICAL MICROSCOPY 153

one method of baking over the other: in actual practice the supe-

rior surface costs less to apply than the inferior and wears longer

and better.

The microscope in the hands of the chemist dealing with this

class of commercial material enables him to rapidly evaluate the

products handled by his firm, to improve faulty products and to

determine whether the materials supplied to the trade are being

properly applied.

Nor must we omit from mention the valuable information we

may obtain from the microscopic study of woods to which fire or

decay-proofing substances have been applied.

I have thus far failed to mention one great industry based

largely upon microscopic control—the Paper Industry. Practically,

an analysis of paper without reference to its ultimate composition

as shown by the microscope is of little or no value. Actually much

paper is manufactured and employed for various purposes without

studies under this instrument. As a necessary consequence we fre-

quently meet with paper-fibre goods manufactured with little regard

for the ends to be attained other than to sell at a profit. A fair

criticism of our American paper products are that they are too good.

That is to say, the quality is higher than need be, and the cost to

the consumer is greater than it should be for many of the objects

to be attained. By that I mean that a less expensive product would

serve equally well and not infrequently better. This is poor busi-

ness and poorer conservation. Let me cite a case in point.

A few years ago a firm manufacturing a product (which must

be nameless, since the investigation was conducted in confidence)

appealed to the laboratory for advice. Their product, and that of

their competitors also, was failing to stand up unaer new condi-

tions of use. In desperation the chemist of the firm wanted to know

whether the microscope would reveal the source of the trouble.

Critical study for a day or two of new and failed material showed

that changes made by the paper firms were probably the cause of the

trouble. A commercially better grade of paper was being supplied.

The matter was taken up with the paper firm. The answer was

quick and to the point. The paper supplied was the highest grade

154 E. M. CHAMOT

that could be produced at that price and further they didn’t pro-

pose to have any men, mere analysts, tell them their business. They

had been manufacturing papers before the questioning men were

born, etc., etc. In fact the same old story and the same old trouble -

with many well-meaning American firms. All you industrial men

have had similar experiences. I need not go further.

A small firm was prevailed upon to make a paper of a kind

which, it was believed, would eliminate certain features which the

microscope seemed to indicate to be the cause, or at least one of the

causes of the troubles. This new paper was then treated in the

proper manner and tested out. The results were so satisfactory that

a contract was placed to take the entire output of the paper firm

with specifications as to the kind of paper needed. The net result

was that a product was obtained in which, not only were the old

defects eliminated, but the cost of production was decreased, the

final profit greatly increased, and the stability of this industry

assured. But I am not sure that the paper firm which lost a large

contract is even today convinced that the new methods of micro-

scopic investigations are of value.

There is little doubt that similar conditions obtain in many of

the other varied paper-fibre industries. Microscopic methods are

the only ones which enable the analyst to identify the nature of

the paper and to indicate its fitness and adaptability for the specific

uses to which it will be put.

The technique for the recognition of the nature of the fibres

present and for their quantitative determinations are fairly well

established and are on the whole, quite satisfactory. But a phase

of microscopic investigation has been neglected. A study of the

finished surface with reference to the uses to which the paper is

to be applied. A study of paper surfaces with vertical illuminator

and with oblique light yields most interesting results. Were these

methods more often employed there would be, in many cases, a

decided modification in certain papers on the market.

The Textile Industries: Like paper, the analyses of textiles

and the recognition of the fibres of commerce are dependent entirely

upon the proper application of microscopic methods. At the present

CHEMICAL MICROSCOPY 155

time no other satisfactory methods are available for differentiating

between the species of fibres employed, the specific treatment they

have received, or the loom arrangement by which the yarns have

been woven into fabrics. In not a few instances the information

thus obtainable may go even farther and disclose the nature of the

method used in dyeing the yarn or the fabric. Uneven adsorption,

variable penetration, etc., are easily recognized. The skilled investi-

gator may go even farther and discriminate between different quali-

ties of the same species.

The technical microscopy of the textile fibres is still in its

infancy, its literature teems with inaccuracies and contradictions.

Too little attention has been devoted to the investigation of the reac-

tions of reagents and the selection of proper differentiating stains,

and the potential possibilities of dark field condensers having very

oblique ray illumination (ultramicroscope) and of luminescence

illumination (ultra violet rays) have not yet received the attention

they deserve.

By way of illustration of what the microscope reveals, I call

your attention to several lantern slides selected to show how neglect

to employ this instrument led to the failure of a manufacturer to

reproduce a fabric which his firm was called upon to manufacture

because of war conditions. The reproduction involved producing

a similar yarn from like fibres, a similar weave in the fabric, and

a similar colored printed pattern. You will see that in no case was

he successful and that his different attempts were a waste of time,

material and energy, since he apparently lacked the fundamental

microscopic information necessary for success.

I trust that in these rambling remarks I may have converted

some skeptics to a belief in the importance of chemical microscopy

in our industries, and may stimulate a wider interest in a branch

of chemical analysis whose value has been greatly underestimated

and whose development has been sadly neglected.

156 E. M. CHAMOT

EXPLANATION OF PLATES

Pirate XI. Fig. 1. Crystallization upon an object slide even so rapid as to

lead to the formation of dendrite crystals rarely results in contact between

the crystal groups. Melting points may often therefore be determined

without the necessity of separating constituents. In the field photo-

graphed are crystals aggregates belonging to the isometric and to the

monoclinic systems.

Pirates XI, XII, XIII, XIV. Figs. 2, 3, 4, 5, 6, 7. A single reagent often gives

very different crystals with different elements. The illustrations show the

crystals produced by the action of potassium mercuric sulphocyante upon

solutions of salts of the elements listed below. Thus, by adding a single

reagent the presence or absence of many substances may be at once

indicated effecting a saving of time, labor and material.

Fig. 2. Reaction with Salts of Copper, giving Cu(CNS). °Hg(CNS)-

°H20, color, greenish yellow.

Fig. 3. With salts of Cobalt, Co(CNS)2°Hg(CNS).; color, deep blue.

Fig. 4. With salts of Zinc, Zn(CNS)2°Hg(CNS).; color, white.

Fig. 5. With salts of Cadmium, Cd(CNS).°Hg(CNS)2; colorless.

Fig. 6. With salts of Lead, Pb(CNS).°Hg(CNS).; colorless.

Fig. 7. With Manganous salts, Mn(CNS)2°Hg(CNS)>; colorless.

Pirate XIV. Fig. 8. Illustrates the amount of space required to perform the

complete analysis of an ordinary commercial alloy (Babbitt). The pho-

tograph is that of the end of a standard 3x1 in. object slide upon which

an actual analysis has been effected. The spots show where the tiny

fragment of alloy was dissolved, reagents applied, and decantation of

drops from one place to another practiced. Since the quantities of material

required are small the time consumed and the amount of reagents required

are reduced to a minimum.

PLATE XV. Figs. 9, 10. Show the application of microscopic methods to the

study of ceramic glazes. Fig. 9 shows a thick glaze improperly bonded

and filled with gas bubbles. Fig. 10 a thin glaze closely adherent to the

porcelain and containing very few gas bubbles.

———— eee ee

PEATESS Ltt

cz)

SOME COMPARISONS BETWEEN NUCLEI OF

NERVE CELLS

By Witttam A. Hirton

Nuclei were compared from a large number of animals, chiefly

invertebrates. In this short paper nuclei alone are drawn. No

special method has been applied at this time. In all cases strong

fixing reagents and strong stains were used, such as Flemming’s

fluid or mercuric cloride. The stain used was hematoxylin in most

cases. Slight differences which one or another reagent gives will

not be entered into at present. The figures were made from prep-

arations which seemed to be typical cells for the animal. In the

methods used no great differences were seen in the appearances of

the nuclei except that by some methods they stained more intensely.

The study is particularly directed towards a comparison of

the distribution of the basichromatin in the nuclei. All drawings

are at the same scale and drawn with a camera lucida with the best

oil immersion objective available.

Many of us appreciate the advances which Biology has made

through the aid of chemical interpretations and physical and chemi-

cal explanations. Some are inclined to belittle pure morphological

studies, but it seems in many cases that we have to come back to

the purely anatomical and morphological investigations in order to

advance. Great attention has been given to the dividing period of

cell activities, but are not the periods between division more import-

ant in many respects? Is it not probable that this is the time when

the nature of the cell is determined? Evidence has gradually been

accumulating to show that the nucleus and the materials within it

are closely related to the cell body and its activities.

The nucleus is probably not concerned with conduction or the

special activities of the neurone, although it may have to do with

nutrition. Even in Protozoa Verworn ’88 (1), denies the nucleus

being the seat of any psychic activity as expressed by motion. The

nucleus seems to have nothing to do directly with irritability ; frag-

(1) Zeit. f. wiss. Zool. llvi.

158 WILLIAM A. HILTON

ments without nuclei respond as readily to stimuli as other parts.

That the nucleus controls or conditions the character of the cell

body seems true to some degree, but just how much, opinions differ.

In those cells which retain their power to divide the chromatin con-

tent remains large, in those which differentiate, this chromatin falls

proportionately, but in some cells it may be restored once more when.

certain stimuli or special nutrition prepares them for division again.

Nerve cells and some others do not as a rule divide after their

mature life is reached, and in these the chromatin content is espe-

cially reduced. E. Rohde ’11 (2) calls attention to the reduction

of chromatin in other than germ cells and compares the loss through

polar body formation to the loss in nerve cells, yolk cells and in

the extreme condition in the red blood cells of mammals.

Although nerve cells do not divide they may give out growths

during this period as shown by G. Marinesco and J. Minera ’14 (3),

who studied a culture of spinal ganglion cells in vitro. The con-

nective tissue cells did grow by multiplication, but the nerve cells

did not divide although they gave out processes. From other evi-

dence it seems possible also that in the adult central nervous sys-

tem there may be growth or change of position of fibers to make

new connections.

M. Muhlmann ’11 (4), has studied the structure of nerve cells

at different ages in mammals. He finds at first a large amount of

chromatin in the nucleus, later it became less, some. goes to form

a sort of nucleolus. The tigroid substance of the cell body gradu-

ally increases. Later Muhlmann ’14 (5) studied the composition

of Nissl’s granules and found that they have a nuclein content

along with globulin, but the nuclein of Nissl’s granules differs from

that of nuclei because the tigroid substance is soluable in alkalis

and the chromatin of the nucleus is not. Just what relation the

lost chromatin of nerve cells bears to the cytoplasm remains to be

clearly determined, but that there is a relation seems clear from

the work of many.

(2) Zeit. f£. wiss. Zool. xceviii.

(3) Anat. anz. 1914.

(4) Arch. mic. Anat. Ixxvii.

(5) Arch. mic. Anat. Ilvi.

COMPARISONS BETWEEN NUCLEI OF NERVE CELLS 159

H. Stauffacher *10 (6) studied the nuclear structure in animal

and plant cells and found the oxychromatic matrix substance of

the nucleolus connected by internal nuclear bridges with the oxy-

chromatin of the nucleus and that, with the oxychromatin of the

cytoplasm. Basichromatin always has an oxychromatin foundation.

Nucleoli arise from oxychromatin and pass to the nucleus and from

there into the cell substance. Nuclein has to do with growth and

metabolism. He does not believe in a nuclear membrane. The

drawings and photographs of Stauffacher look very convincing. I

have found similar conditions in some germ cells. J. Schaxel ’10

(7) speaks of the regulative emmission of chromatin to the cell

from the nucleus. W. Knoll 710 (8) has studied the leucytes in

bone marrow. Nuclear bridges connect contractile parts of the

nucleus with contractile parts of the cytoplasm. There are many

other papers with similar evidence which suggest either a close

chemical interrelationship or definite organic connection between

cytoplasm and nucleus.

That nuclei may receive again from the. cytoplasm, or that the

cell body aids the nucleus in the growth of its chromatin seems clear

from the recent work of V. Danchakoff ‘16 (9). “Accumulations

of basophilic substance are found in the cytoplasm * * *. The small

achromatic nucleus of the mature egg both after fertilization and

at the beginning of parthenogenesis seem to exert a strong attrac-

tion on the basophilic accumulations found in the cytoplasm. This

substance is soon displaced and localized around the nucleus. At

the same time a radiation around the nucleus is perceptible, which

is apparently the expression of the flowing of the basophilic sub-

stance towards the nucleus * * *. ‘The close relation bétween the

true chromatin bodies within the nucleus and the basophilic chro-

matic substance in the cytoplasm is easily traced.”

In a study of a large number of nerve nuclei such as shown

in figs. 1 to 17 and fig. 22, it is found that in most nuclei there is

but a scanty distribution of nuclear material which is proportionately

(6) Zeit. f. wiss. Zool. xcv.

(7) Verh. viii Intnat. Zool. Kong. zu Graz. 1910,

(8) Zeit. wiss. Zool. xcv.

(9) Jour. morph. vol. xxvii, no. 3,

160 WILLIAM A. HILTON

less in the larger nuclei of cells that have very large-cell bodies.

In most cells there is a well-marked, usually clear nucleolus. In many

there are many fine strands of linin visible, usually radiating from

the center of the nucleus. The smallest animals have the smallest

cells and these have the smallest nuclei. Very often the smallest

nuclei do not look much like those of nerve cells, such as those of

Podura (fig. 13) and Aphorura (fig. 12), seem more like neuro-

blasts or young cells although their chromatin content is small. In

the larger animals there was considerable variation noted in cells

from the same animal, such as in L. grandis, fig. 6, Trantula fig. 10,

Corydilis, fig. 16.

In all the preparations probably the more granular nuclei are

those from neuroblasts and a few of the clear nuclei may be of

neurogolia cells. In the nerve nuclei of a young salamander

Desmognathus, fig. 22, the chromatin is very abundant and the cells

with such nuclei are capable of active multiplication. In figs. 23

and 24 from muscle and connective tissue cells, the scanty chro-

matin would be insufficient for immediate division. Figs. 18 to

21 are from another salamander, a larval Amblystoma. Fig. 18

from the retina might soon have more chromatin and divide, one

of the epithelial cell nuclei of fig. 19 could soon divide and fig. 20

from the blood has a mass of chromatin and the muscle nucleus of

fig. 21, is not near division.

No more than a general idea of the distribution of the chro-

matin in the nuclei of nerve cells can be learned from these figures.

A general type of nucleus shows fairly well in all but the smallest

and youngest specimens. The type is that usually recognized for

the nerve cells of vertebrates, that is a large rather clear nucleus

with a nucleolus which is also rather clear. There seems to be

nothing especially peculiar to nerve cell nuclei which would pre-

vent the cells of the adult from receiving a stimulus of some sort

to increase the chromatin of the nucleus and allow further division

of the cells.

Whether the arrangement of the chromatin means anything or

not in determining the nature of the cells cannot be told from these

preparations, but I do not doubt that this as well as any particular

COMPARISONS BETWEEN NUCLEI OF NERVE CELLS 161

or peculiar arrangement of the nuclear parts may be of importance

at one time or another in determining the nature of cytoplasmic

manifestations.

Zoological Laboratory of Pomona College,

Claremont, Califorma.

162 WILLIAM A, HILTON

EXPLANATION OF FIGURES

All figures drawn by the aid of a camera lucida by means of an oil

immersion objective. All enlarged eight hundred diameters.

Fig. 1. Nerve nuclei from the starfish, Orthasterias.

Fig. 2. Nerve nuclei from the starfish, Piaster capitatus.

Fig. 3. Nerve nuclei from the crustacean, Nebalia.

Fig. 4. Nerve nuclei from the crustacean isopod, Lygida.

Fig. 5. Nerve nuclei from the crustacean isopod, Lygida 4mm. long young.

Fig. 6. Nerve nuclei from the crustacean decapod, Loxorhynchus grandis.

Fig. 7. Nerve nuclei from the pycnogonid, Lecythorhynchus marginatus.

Fig. 8. Nerve nuclei from the whip-scorpion, Tythreus pentapelltis.

Fig. 9. Nerve nuclei from the pseudoscorpion, Chelifer.

Fig. 10. Nerve nuclei from the tarantula spider.

Fig. 11. Nerve nuclei from the thysanuran insect, Campodea.

Fig. 12. Nerve nuclei from the collembolan insect, Aphorura.

Fig. 13. Nerve nuclei from the collembolan insect, Podura.

Fig. 14. Nerve nuclei from the collembolan insect, Sminthurus.

Fig. 15. Nerve nuclei from the thysanuran insect, Machilis.

Fig. 16. Nerve nuclei from the larval insect, Corydalis.

Fig. 17. Nerve nuclei from the ascidian, Tunica.

Fig. 18. Nucleus from one of the retinal cells of the larval salamander

Amblystoma 15 mm. long.

Fig. 19. Nuclei from epithelial cells of larval Amblystoma, 15 mm. long.

Fig. 20. Nucleus from the blood of a 15 mm. larval Amblystoma.

Fig. 21. Nucleus from the muscle of a 15 mm. larval Amblystoma.

Fig. 22. Nuclei from brain of a young salamander, Desmognathus.

Fig. 23. Nucleus from striated muscle from young Desmognathus.

Fig. 24. Nucleus from connective tissue from young Desmognathus.

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DEPARTMENT OF NOTES, REVIEWS, ETC.

It is the purpose, in this department, to present from time to time brief original

notes, both of methods of work and of results, by members of the Society. All

members are invited to submit such items. In the absence of these there will be given a

few brief abstracts of recent work of more general interest to students and teachers..

There will be no attempt to make these abstracts exhaustive. They will illustrate progress

without attempting to define it, and will thus give to the teacher current illustrations, and

to the isolated student suggestions of suitable fields of investigation.—[Editor.]

ese

NOTES ON THE CULTURING OF MICROSCOPIC ORGANISMS FOR THE

ZOOLOGICAL LABORATORY

General.—The culture of micro-organisms, especially the Pro-

tozoa, is an old topic among biologists. Leeuwenhoek in his quaint

language records not only the finding of Protozoa in various natural

waters but also in infusions of pepper. He was thus the first to

make cultures of the Protozoa. Leeuwenhoek’s cultures were largely

due to a series of accidents or to haphazard experiments. Many

zoologists of today use this haphazard method of making cultures.

It is the usual practice to collect a mass of vegetable material from

a pond, stream or other body of water. This mass is placed in

almost any kind of a dish, and left in strong light or in darkness,

covered or uncovered as long as the preparator can stand the odor.

From these haphazard methods he gets some surprising results

but quite frequently not at the time when wanted. If he has made

these cultures with some particular protozoan in mind, he is quite

likely to find the species mixed (if he finds it at all) with several

other species. When through using these cultures or when they

are spent he throws them out and makes more in the same haphazard

fashion.

These have been and are yet to a large extent the methods of

culturing Protozoa as practiced by the zoologist. Bacteriologists,

however, have developed very careful methods of culturing bacteria

and some few species of Protozoa. They make use of media con-

taining known and measured ingredients, kept under controlled con-

ditions. The response of the organisms to particular media and

to particular conditions is characteristic and the habit of growth

164 AMERICAN MICROSCOPICAL SOCIETY

is an aid in diagnosis of the species under observation. The bacteri-

ologist cannot use haphazard methods and secure results. Botanists

also are making use of exact methods in the culturing of plant organ-

isms as alge, fungi and even higher green plants. The use of exact

methods by botanists has been promoted by the fact that many of

them are trained in bacteriological methods and in the exact methods

of plant physiology. Their research in the life histories and in the

metabolism of lower plants has demanded the use of exact methods.

The zoologist, too, must come to the use of quantitative methods

in the culturing of many of the lower forms of laboratory animals.

He is, however, more or less dependent on the bacteriologist and

the botanist to show him the methods of rearing the food organisms

for his Protozoa or his worms. The zodlogist can, indeed, help

himself very greatly if he will learn to use the cultural methods of

the bacteriologist and the botanist. It is not enough that some

zodlogists make use of these methods but having worked out methods

that are successful they should by early publication make their meth-

ods available to others. The writer has no sympathy with the disin-

clination of some workers to make their methods known. There is

a real need among amateur zoologists, teachers and investigators

for a greater knowledge of technical methods and such informa-

tion ought to be made readily available.

Some criticisms of the old methods may be given and some

suggestions offered concerning food supply, dishes, light, inocula-

tion and culture media.

Food Supply.—One of the great faults of the old methods of

culturing micro-organisms was in the failure to maintain a stock of

food materials in cultures that had begun to yield. When the culture

was made it was heaped high with food materials which were never

replaced as they became exhausted by the action of bacteria. With

depletion of the organic matter the crop of bacteria or of unicellular

alge which furnished pasturage for many of the Protozoa rapidly

diminished in numbers and the Protozoa decreased to the vanishing

point. For this reason it has been the custom to bring in cultures

every few days in the hopes of having one culture ready when

wanted. It should be borne in mind that if the number of animals

in a culture is to be maintained their food supply must be replen-

NOTES, REVIEWS, ETC. 165

ished. This can be done by adding hay, manure from various

sources or manure solution, or other vegetable or organic material

which does not quickly become acid in the process of decay. Rapid

souring of culture medium is. fatal to most Protozoa. The writer

has never been very successful in growing Protozoa in media con-

taining much of such carbohydrates as occur in wheat.

Culture Dishes ——Even the kind of a dish in which the culture

is maintained is of importance. The best dishes are shallow and

have a diameter considerably greater than their depth. Covered

glass dishes ca 200 mm. in diameter by 80 mm. in depth and known

as bacteria dishes or culture dishes are excellent but not indispen-

sable. The writer has used Stender dishes 50 mm. in diameter by

30 mm. in depth with success although a small culture is more diffi-

cult to maintain than a larger one. Fruit jars, battery jars and

even aquarium jars are satisfactory if the liquid of the culture is

not too deep. Earthenware jars and crocks are satisfactory for

forms preferring the dark. Keep all cultures covered.

Light.—As to light, diffuse light or even darkness may be used

for most forms except those which feed on a green or blue-green

plant or which like Euglena need light for photosynthesis. Even in

these cases the diffuse light and more even temperature of a north

window are to be preferred to direct sunlight and the accompany-

ing violent fluctuations of temperature.

Inoculation—If in the wild cultures the desired organisms

appear in numbers new cultures may be inoculated with a small

quantity of material from the old culture, taking precaution to add

the food organism also and to avoid contamination with other organ-

isms. Thus one may become reasonably free from the necessity

of frequent trips to pond or stream to collect vegetation for cul-

tures. The culturist should study the food habits of the organism

and strive in preparing the new culture to provide the proper medium

for the development of the food organism and then he must pro-

vide proper conditions of light, temperature and gaseous exchange.

If by observation or experiment he can determine the ingredients

to be used in his culture and the conditions for the growth of the

food organisms he should be able to maintain cultures almost

indefinitely.

166 AMERICAN MICROSCOPICAL SOCIETY

Culture Media—Various culture media have been suggested for

general and special Protozoa cultures. The writer has confined his

efforts to a few media. One of the most successful of these for

general cultures is made of sterile hay and filtered tap-water. To

prepare this medium timothy hay is made into small compact bun-

dles which are tightly wrapped in several layers of cheese cloth.

The bundles are sterilized once in a steam autoclave at 15 to 17 Ibs.

pressure for 15 minutes or more, then dried over a steam radiator

or in a drying oven. This sterilization is not intended to kill bac-

teria or spores of fungi but to kill encysted Protozoa if any be

present. The tap-water is filtered through a paper filter, preferably

‘nto sterile dishes. The idea here is to free the water of any Pro-

tozoa which are always present in tap-water of this city but not

necessarily to get rid of the bacteria. Ten grams of the sterile hay

to two liters of filtered tap-water makes a very satisfactory medium

for many kinds of Protozoa. These proportions, however, can be

varied considerably. When a culture has been started in this medium

by inoculation from a previous culture it may be maintained by the

addition of one or two grams of sterile hay per week and as much

filtered water as is necessary to maintain the original level of liquid.

Boiled hay solution has been tried repeatedly and has never

given good results, apparently because of the development of toxic

substances. It usually soon acquires an odor somewhat resembling

butyric acid and when in this condition if Paramecia are placed in

it they die in a very few minutes.

Manure solution made by boiling fresh horse or cow manure

or human feces in tap-water and diluting with filtered tap-water _

has given excellent results in culturing Euglena and some unicellular

green alge as Scenedesmus. Any of these may serve as food organ-

isms for some of the Protozoa. Unfortunately the proportions in

which the manure solutions were made have not been kept. It has,

however, been the experience of the writer that the proportions

may be widely varied. Manure solutions may be added to the hay

cultures with beneficial results.

The writer has found the filtered tap-water superior to rain-

water which is lacking in many of the salts necessary for most

organisms. Distilled water has never been used because it has been

NOTES, REVIEWS, ETC. 167

simpler to use the filtered tap-water than to add salts in the proper

proportions to the distilled water. Boiled tap-water is inferior to

filtered tap-water probably because boiling precipitates a large pro-

portion of the salts and drives off the oxygen and carbon dioxide.

In the event that the tap-water has been treated with chlorine it

may be necessary to use filtered pond or stream water or spring

water.

Culture of Amaba.—Two of the writer’s students, Miss

Knisely and Mr. Welch, have worked out methods of culturing

ameba on solid media. The results of this work were published in

in these Transactions, vol. 34, pp. 21-25.

The writer’s work in culturing ameba has been incidental to

the securing of materials for class use and some of the results

attained have been more or less accidental. Ameba cultures have

been secured in many ways of which only a few are related.

During the summer of 1916 small amebz, probably soil amebz,

occurred in great abundance in a culture made from unwashed rad-

ishes pulled from a damp garden. No efforts were made to main-

tain the culture and the amebz soon disappeared.

On Aug. 29, 1916, some lily-pads and pond water were taken

from First Sister Lake, near Ann Arbor. This culture yielded

among other Protozoa a fair number of large amebe. On Dec. 29,

1916, a sterile-hay-filtered-tap-water culture was made up in a bac-

teria dish and some scum from the preceding culture was added.

In this culture a slender diatom and the ameba established them-

selves and have thriven to date. This ameba is a large rounded

sluggish form measuring nearly 100 microns in extreme dimension.

Its surface is wrinkled. It puts forth few pseudopodia, has a thick

hyaline ectosarc, finely granular entosarc, large contractile vacuole,

and a well-defined ellipsoidal nucleus within a wider clear area.

Specimens have usually been very numerous in this culture and

since there have been very few other organisms present it has been

excellent for student use. To this culture were added at intervals

of one to several weeks a few grams of sterile hay until June 21,

1917. At this time the culture was in good condition. The diatoms

were very numerous on the bottom of the dish where the sluggish

_amebze were abundant. There were also many more active amebz

168 AMERICAN MICROSCOPICAL SOCIETY

resembling A. proteus. From June 21 until Aug. 30 no hay was

added to this culture and it was not examined meanwhile. On the

latter date amebz of the above mentioned kinds were found but

not abundantly. About 2 grams of hay were added Aug. 30 and on

Sept. 19 both kinds of amebe were quite abundant.

On Jan. 9, 1917, a subculture in a sterile-hay-tap-water medium

was made from the culture above mentioned. For a long time it

yielded amebe altho less abundantly than the parent colony, but on

June 21 it was producing quantities of diatoms upon which large

numbers of the sluggish ameba were feeding. This culture had

more hay than the parent culture. It was uncared for from June

21 until Aug. 30 when a few amebe were found. Hay was added

Aug. 30 and on Sept. 19 the amebe were more abundant.

The above cultures have always been kept near a north window

in diffuse light and have always been kept covered with a glass

plate. Recent comparison of these amebe with amebe secured on

Sept. 12, 1917, from the original source of these cultures shows

that they have not changed in form or size.

Culture of Euglena—Euglena occurs most abundantly in foul

waters and is well known as an inhabitant of sewage-polluted waters.

Euglena cultures have been maintained in luxuriance with almost no

attention on the sterile-hay-filtered-tap-water medium for more than

a year in the diffuse light of a north window. They also have been

transplanted into manure solutions where they have produced a dense

green scum over the surface of the water and down the sides of

the jar and as far above the water on the sides of the jar as the

film of moisture extended. After a time large numbers of these

Euglenz become encysted. It has been found by Dr. A. F. Shull

of our laboratory that such Euglenze soon emerge from the cyst

if placed in a fresh manure solution. Thus the providing of Euglena

for class use which formerly was the bugbear of the general zoology

course is now a very simple matter.

Culture of Tetrastemma.—A fresh water nemertean worm

which resembles Tetrastemma and has provisionally been assigned

to that genus has appeared in cultures taken from First Sister Lake,

near Ann Arbor, on July 12, 1916. Until September, 1916, this

culture was maintained in a covered pint fruit jar, then was put in

NOTES, REVIEWS, ETC. 169

a bacteria dish with filtered tap-water and some sterile hay. Tetra-

stemma was first noted in November. The culture was maintained

by adding a few grams of sterile hay at intervals of one or more

weeks. During most of the winter this organism was abundant.

On March 23, 1917, this worm was present in large numbers, but

by June 21 its numbers were greatly reduced. On Aug. 30 a single

specimen was found in a subculture.

Culture of Pristina—In the culture in which Tetrastemma

appeared and at the same time the oligochete worm, Pristina sp.

appeared and was maintained in considerable abundance until late

February and although on March 23 it was on the decline it was still

present in cultures on June 21. Subcultures were made from time

to time during the winter in which this worm thrived. The species

has disappeared in these cultures during the summer. The cultures

of Tetrastemma and of Pristina were maintained just as Paramecium

cultures were maintained. In fact during the greater part of the

time Paramecia were very numerous in all the cultures of these

worms. .

University of Michigan, GeorGE R. LaRue.

Ann Arbor, Mich.

EFFECTS OF THYROID ON PARAMECIUM

Shumway (J. Exp. Zool., Apr., 1917) cites a number of inter-

esting results from culturing Paramecium in emulsions and suspen-

sions of thyroid. Among these are,—that the division rate is in-

creased 65% over the controls; that this acceleration of rate is

greatest at the time when the controls themselves are dividing most

rapidly, and hence the curves show the same general rhythms as

those of the controls; that the number of contractile vacuoles

increases from two to three, indicating a disturbance of the excre-

tory function; that large vocuolation takes place in the protoplasm,

which is believed to have an excretory significance, altho the vacuoles

do not pulsate. They were often observed to burst, freeing their

contents internally.

170 AMERICAN MICROSCOPICAL SOCIETY

Of all the hormone-secreting glands the thyroid is the only one

to produce the above results. Boiling does not destroy the activity

of the material. Iodine and iodothyrin do not produce it. )

When the treatment is discontinued the strain of Paramecium

reverts to the division rate of the controls. The other mutations

brought about by the feeding do not seem to persist.

ARE CONJUGATION AND ENCYSTMENT NECESSARY ?

Mast (J. Exp. Zool., July, 1917) concludes that neither conju-

gation nor encystment is necessary for the continued existence of

Didinium. By starting new groups of lines from old ones both

near and remote from encystment and propagating these in parallel

series, it seems that these data have no constant or appreciable

effect on fission rate, or on variations in this rate. In these experi-

ments a group was carried an average of 1646 generations without

encystment. In one group of lines, 721 generations after conju-

gation and 196 generations after encystment, a mutation suddenly

appeared in which the rate of division for 315 days was more rapid

than the slower lines in the ratio of 836 generations to 634.

EXTRA CONTRACTILE VACUOLES IN PARAMECIUM

Hance (J. Exp. Zool., July, 1917) gives the account of the

origin of a race of Paramecium with supernumerary vacuoles, appar-

ently arising thru subjection to temperatures higher than usual. The

race was unusually strong, large, and resistant to extreme conditions.

The extra vacuoles, ranging as high as five, usually occurred in

the posterior end of the cell. They may increase during the vege-

tative life of the individual; and the number is modified by the rate

of division, the age of the culture medium, and the presence of kata-

bolic products in the environment. The author holds that the “poten-

tiality for this organ has not been lost, tho the extra vacuoles may

not actually appear for several generations.”’

PRESERVING FISH WITHOUT ICE

Sherman’s Fish Sterilizing Co. owns a process for preserving

fish without ice (Pacific Fisherman, July, 1917) which is claimed

NOTES, REVIEWS, ETC. 171

to keep the fish fresh and firm with all the original flavor for a fort-

night or longer. The process takes about 3 hours. The fish are

first put in a cooling tank filled with water and brought to a low

temperature, where they are left for one-half hour. It is next placed

in a tank of cold sea water or fresh water strengthened with salt.

In order to prevent freezing in the brine the solution is kept agi-

‘tated by pumping. It is claimed that the low temperature salt solu-

tion closes up the surface pores, preventing saturation and also

acting as an antiseptic protection on the outside.

It is claimed that the process is cheap in installation and opera-

tion and will enable the small producer to get his catch on the market

in good condition, thus aiding increased production and safe mar-

keting.

CYTOLOGICAL CHANGES ACCOMPANYING DESICCATION IN ROTIFERS

Hickernell (Biol. Bul. June, 1917) gives a most interesting

account of the desiccation changes in the rotifer Philodina roseola,

with special reference to cell behavior. Attention is called to the

fact that rotifers are resistant not merely to drying. They are resist-

ent to great cold (even — 40° F does not kill them), to great heat,

to strong brine, or to rapid changes from one to another of these

unfavorable conditions. Murray has summarized this hardiness

thus: “Such is the vitality of these little animals [certain south

polar Bdelloids] that they can endure being taken from ice at a minus

temperature, thawed, dried, and subjected to a temperature not

very far short of the boiling point, all within a few hours.”

The structural changes in Philodina during desiccation are

summarized as follows: The animal as a whole contracts into a

form similar to that of extreme contraction in life; no special pro-

tecting membrane is formed,—contrary to the common view; the

tissues and cells maintain their identity during the drying process;

the cytoplasm becomes more dense, and the chromatic part of the

nucleus changes from a single large karyosome, separated from a

distinct nuclear membrane by a clear space, to chromatic network at

the periphery with the clear space internal.

172 AMERICAN MICROSCOPICAL SOCIETY

The author believes that some metabolism goes on during the

dry condition. The evidence for this lies in the fact that food gran-

ules in intestinal cells disappear during desiccation in a manner |

similar to that seen in starving specimens.

It is suggested that the movement of the chromatin to the peri-

phery of the nucleus during desiccation facilitates cell oxidations

during the period. Their change in position takes place at the very

beginning of the drying process. On recovery from desiccation the

changes are the reverse of those described above.

The acceleration of reproductive activity often noted in rotifers

soon after recovering from drying is credited to an increase in

ovarian nuclei that occurs during the period of recovery. When

death results from desiccation any of the following causes may oper-

ate:—mechanical injury due to too rapid drying; starvation due

from a lack of reserve food material; poisonous effects of metabolic

products; insufficient time during early drying to allow the nucleo-

cytoplasmic reorganization.

THE FUNDUS. OCULI OF BIRDS

In a most attractive atlas Dr. Wood has presented the results

of his studies upon representatives of all the leading orders of birds,

including more than 100 species. He rightly takes the view that a

group of animals with such highly developed and varied vision as

birds offers a most fruitful field for the study of all the elements

entering into the structure and activity of the eyes.

His method provides for study with the self-luminous ophthal-

moscope, macroscopic examination of excised eyes, and microscopic

examination of special portions. Owing to the fact that the mydri-

atics usually used for man have little or no direct effect on the

sphincter iris musculature of the bird, it was found necessary to

secure dilation by the use of such agents as galvanism, nicotine,

curare, and of drugs that render the bird unconscious without actu-

ally killing it. It was also found that maximum dilation is to be

had a few moments before and after the death of birds, where these

are being killed for detailed studies of eye-structure.

NOTES, REVIEWS, ETC. 173

The evidences are abundantly manifest of close and effective

cooperation between the author and the artist, Mr. Arthur Head of

London, in the remarkable series of colored plates of some 58

species of birds and 3 species of reptiles, not to mention more than

100 black and white drawings.

The chief variable features of the fundus oculi capable of study

by their combination of methods are:—the color and form of the

internal background ; the shape, size, and degree of development of

the pecten ; the optic nerve entrance; the choroidal and retinal blood

vessels ; the details of distribution of the retinal elements, including

the opaque nerve fibres; the shape and size of the optic discs or

areas of acute vision.

Some of the outstanding conclusions of the author are:—

1. Such examination of the fundus as has been indicated fur-

nish very different and characteristic fundus pictures for different

groups of birds; sufficiently so in many cases for the immediate

recognition of the species. These fundus pictures will rank with

other taxonomic indicia in classification. The author is able to sug-

gest certain broad classifications of fundi.

2. There is. great variety among birds in respect to the size,

shape, and position of the areas of distinct vision. This localization

corresponds strikingly with the habitats and habits of the birds,—

especially to manner of getting food, escape from enemies, migra-

tion, reproduction, etc.

3. Domestication or prolonged captivity seems to bring changes

in the eye of birds. These abnormalities include choroidal diseases,

opaqueness of media, less constancy in many of the variable ele-

ments of the fundus.

4. Study of the fundus of birds in comparison with those of

other vertebrates may be expected to throw light upon the origin

and kinship of the group. |

The chapters are:—1. Introduction; 2. Summary of Conclu-

sions; 3. Collection, Selection, and Preparation of Material; 4.

Review of Anatomy and Physiology of the Fundus Organs; 5. Oph-.

thalmoscopy ; 6. Ophthalmoscopy of the Fundus in Living Birds;

7. Macroscopic Appearances; 8. Effects of Domestication on the

174 AMERICAN MICROSCOPICAL SOCIETY

Fundus Oculi; 9. Appearance of the Fundus Oculi in Various Orders

of Birds; 10. Classification of Ocular Fundi; 11. Ocular Fundus

in Relation to Classification of Aves; 12. Relation of Reptilian to.

Avian Fundi; 13. Atlas of Colored Plates of Fundus Oculi in Birds.

The mechanical and artistic qualities of the book are of the

first rank. It is an ungracious thing to refer to small matters of

incompleteness where the larger things are so painstakingly and

admirably cared for. There is, however, incompleteness and lack

of uniformity in the use of subdivision headings and numerals in

Chapter VI particularly, and a lack of correspondence between these

and the table of contents.

The Fundus Oculi of Birds, Especially as Viewed by the Ophthalmoscope, by Casey

Albert Wood. Illustrated by 145 text drawings, and 61 colored plates. The Lakeside

Press, Chicago, 1917. Price $15.

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ORGANIZED 1878

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1917

VOLUME XXXVI

TRANSACTIONS

OF THE

AMERICAN:

MICROSCOPICAL

SOCIETY

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TRANSACTIONS

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ORGANIZED 1878 INCORPORATED 1891

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TABLE OF CONTENTS .

FOR VOLUME XXXVI, NUMBER 4, OCTOBER, 1917

Rh opsie of the Blue-Green Alge—Myxophyceex, with Plates XVII—XXIX,

79 Micures, bys J osepnine:; Fr TuGeniiiiy vss bos soe ole tee ta hab euie ee oes 179

Notes and Reviews: Technical Methods, with Plate XXX, by H. E. Met-

calf; Further Notes on Rearing Volvox, George R. LaRue; Pyridine Sil-

ver for Bone Sections, by Dr. J. S. Foote; Entomological Notes, by Paul

S. Welch; Photomicrographs in Color; a Short Method of Preparing His-

tological Material, by H. R. Eggleston; Organic Evolution (Lull) ; Origin

and Evolution of Life (Osborn) ; The Microscope (Gage).

TECPOL ER ay Hed lala icia share Gite Moat, jibe Wha kiss © AD wrela a bi dlacerabagett od eats et a Pa 267

Te ee A ee Se ae Ms ir Ea ee ae Ee Set he ire Ore atk ate a ee Le 285

NotTicE TO MEMBERS

The Annual Business Meeting of the American Microscopical

Society for 1917, will occur in affliation with the A. A. A. S. at

Pittsburg, Pa., during Convocation Week, as follows:

Friday, Dec. 28, 12:30, Executive Committee will lunch together

at the Hotel Schenley.

Saturday, Dec. 29, 12:15 P. M., the Business Meeting will occur

in Room 304 Applied Industries Building, following meet-

ing of Ecological Society of America.

All members present at the Pittsburg meeting are requested to

participate.

T. W. GALLoway,

Secretary-Editor.

(This Number was issued on December 18, 1917.)

TRANSACTIONS

American Microscopical Society

(Published in Quarterly Installments)

Vol. XXXVI OCTOBER, 1917 No, 4

SUMMARIES IN MICRO-BIOLOGY

The Secretary has been seeking to secure for this Journal and its Department of

Summaries, a series of papers from biologists dealing with the chief groups of microscopic

plants and animals. It has not been the purpose to present a complete survey of any of the

groups. The wish has been rather to bring together in one article a statement of the fol-

lowing things:—general biology, the method of finding, the methods of capture and of keep-

ing alive and cultivating in the laboratory; how best to study; the general technic; the most

accessible literature; and a brief outline of the classification, with keys for the identification

of at least the more representative genera and species of the micro-organisms likely to be

found by the beginning students in the United States.

It has been felt that the getting together of such data as this, while not a contribution

to science, would be a contribution especially to isolated workers and to teachers and stu-

dents in the high schools and smaller colleges.

Papers have already appeared treating the aquatic Oligochetes, the Melanconiales, the

Rusts, the Black Moulds, the Powdery Mildews, the Cephaline Gregarines, the Conjugate

Algz, and the Free-swimming, Fresh-water Entomostraca. The following is the ninth

paper of the series. It is proposed to have such synopses from time to time until the

more common American species of such groups as the following have been covered: The

non-conjugating Green Alge, Downy Mildews, Yeasts, other Hyphomycetes, Smuts,

Rhizopods, Infusoria, Turbellaria, Bryozoa, Water Mites, etc. [Editor.]

SYNOPSIS OF THE BLUE-GREEN ALG#

—MYXOPHYCE/E

By JosEPHINE E, TiLpENn

The Myxophycee (Cyanophycez, Schizophycee) in structure

and function are the simplest and most primitive of Alge. They

are deserving of study for several reasons. In some ways at least

they appear to be somewhat closely related to the Bacteria. Having

the advantage of larger size and more definite structure their life

histories and habitats are of interest to students of bacteriology and

medicine. They are of economic importance in connection with the

problem of pure water supplies for cities. Geologists now consider

that large existing deposits of marl, travertine and silicious sinter are

the result of the work of these minute plants. To students and

180 JOSEPHINE E. TILDEN

teachers of botany the study of the blue-green alge forms a useful

introduction to the higher forms.

Collecting Blue-green Alge

The common Blue-green alge are to be searched for in ponds,

in small pools on swampy ground or along roadsides, on moist and

dripping rocks, at the edges of lakes and rivers. Along the seashore

they will be found in tide-pools, on rocks, and clothing the fronds

of larger alge. Some of these alge are able to live in hot waters

and a study of the vegetation of hot springs and geysers forms an

interesting problem.

It is usually not difficult to find sufficient material for class use

even in the winter months, in springs, on dripping rocks, or at any

rate in tanks and vessels in green houses. It is, however, easier to

preserve specimens when they are found in abundance and so have

them on hand when needed. To collect them one needs some wide-

mouthed vials, a knife or scalpel for scraping damp boards and rocks,

a spoon for dipping minute floating forms from the water. If one

desires to preserve the specimens for future microscopic work, they

should be immediately placed in vials containing a 2 or 3 per cent

formaline solution. A descriptive label should be inserted in each

vial, containing date of collection and notes on habitat, color, form,

locality, etc. On the other hand, specimens designed to be kept alive

in the laboratory for class work should be transferred as soon as pos-

sible from the collecting vials to broad glass tanks or jars which

should stand in or near a north window where they will not receive

too much sunlight. It is well to cover these jars with glass plates.

If one desires to determine the species of these algz it is nec-

essary to possess an eyepiece micrometer or a micrometer disc

(which fits inside standard eyepieces).

Structure

During the last few years a number of investigators have given

their attention to determining the nature of the protoplast of the

Blue-green alge. The majority of them now consider that there is

a definite cell wall, an “incipient nucleus,” cytoplasm, and three main

pigments.

SYNOPSIS OF THE BLUE-GREEN ALG#—MYXOPHYCEZ 181

The cells in this group consist of a protoplasmic mass sur-

rounded by a wall or membrane and containing a single “incipient

nucleus.” Protoplasmic connections between cells have been dem-

onstrated in certain cases. In an actively growing condition the

protoplasm fills the cell. In long continued dryness it contracts away

from the wall. Young growing cells contain many granules, but

when older their contents become homogeneous. The cytoplasm con-

tains chlorophyll, phycocyanin, and carotin (pigments), several kinds

of granules, a slime substance, vacuoles, oil-drops, glycogen and

sugar. Probably sugar is the first product of photosynthesis. The

sugar is changed into glycogen as a carbohydrate reserve. Starch is

not found in the Blue-green alge. The cyanophycin granules are a

product of assimilation and represent reserve albumen.

True chromatophores are not present. The green chlorophyll

combined with the bluish phycocyanin give the characteristic blue-

green color to the cells of these plants. Red or pink tints are due

to the predominance of carotin. The chlorophyll and carotin appear

to exist in the form of very minute granules, while the phycocyanin

occurs in a diffuse state. To the age of the cell and the color of the

sheath is due the difference in the color of the protoplasm in different

parts of the same plant. It is generally more greenish in young parts

which are covered by a colorless sheath, while in older ones it is olive

or yellowish-green. Sometimes the terminal cells of a trichome, in

certain genera, show a rose-colored tint.

By the use of various stains it has been found that each cell con-

tains a single nucleus, the so-called “central body.” It is much sim-

pler than the nucleus of other plants, since it possesses neither a

nucleolus nor a nuclear membrane, and the best descriptive term now

in use seems to be “incipient nucleus.” The resting nucleus consists

of a slightly staining ground mass in which numerous granules lie

imbedded. These granules are not true chromatin but nothing much

so far is known about them. During cell division the incipient nuc-

leus seems to divide without mitosis. It constricts and the new cell-

wall first appears as a ring midway between the daughter-nuclei.

This becomes gradually filled in towards the center until the wall

between the two daughter-cells is complete.

182 JOSEPHINE E. TILDEN

At first the cell wall is composed principally of cellulose. Later

it becomes changed in character, resembling fungus-cellulose or the

cuticle of higher plants. It may in some cases contain silica and in

others chitin. The outer portion is mucilaginous. Blue-green alge

in all cases secrete mucilage and are usually found imbedded in

masses of mucus. In filamentous plants the mucus forms an en-

veloping “sheath” which may dissolve and become invisible, or in

other cases may undergo a toughening process. Such a sheath is a

secretion of the cells of the plant.

The heterocyst is a special cell usually larger than the ordinary

cells. It is formed from a vegetative cell by the breaking down of

the incipient nucleus, a loss of the granular character of the cell

contents, and the thickening of the cellulose wall. The position of

the heterocyst in the trichome varies, sometimes it being confined to

the end of the row of cells, and in other cases it lies between two ordi-

nary cells. Although usually colorless, the watery contents of the

heterocyst sometimes have a more or less decided yellowish tint.

Heterocysts are usually solitary but they may occur in short chains.

While it is a fact that the function of the heterocyst is at present

not at all understood, it is doubtless concerned with the breaking up

of the trichome into hormogones. By some it is considered to act

as a storehouse for reserve substances.

In the unicellular forms the terms “plant” and “cell” are synony-

mous. Therefore when the cell divides the result is not growth but

multiplication, for two distinct new plants have been formed. In all

other cases the cells cling together after division and are arranged

in rows or strings. The “plant,” then, consists of a row of cells.

The entire number of cells in a blue-green multicellular plant (includ-

ing heterocysts and gonidia if present) taken together, is known as

the “trichome.” In many genera the trichome is surrounded by a

gelatinous sheath or envelope. The trichome together with its sheath

is called the “filament.”

In the multicellular forms the plant grows by the transverse

division either of all the cells, as in Nostoc, or of a group of cells

occupying a definite position, as in Rivularia. Ordinarily division

takes place as soon as the cell has attained its maximum size; but

under favorable conditions secondary divisions occur in the young

SYNOPSIS OF THE BLUE-GREEN ALG#—MYXOPHYCEZ 183

cells and when this happens the cells are much shorter than broad,

that is, the cross walls appear quite close together.

Reproduction

In the unicellular forms multiplication is the result of simple

cell division. In some genera gonidia, formed within gonidangia,

occur. |

The multicellular Blue-green alge reproduce commonly by

means of hormogones. A hormogone is a short chain or row of cells

broken off from a mature plant. For a short time it has the power

of motion. Its cells rapidly divide and thus a new plant is developed.

A second method of reproduction in the multicellular forms,

is by means of gonidia. These are either formed within gonidangia,

or, as is most commonly the case, developed from vegetative cells

into thick-walled resting cells.

Water-Supply Algz

A number of species belonging to the Blue-green Algz are of

interest because of their effect upon drinking water.

Some of these algz possess a natural odor and taste, due to the

presence of an oil, which upon being liberated produces a most dis-

agreeable result. The odor is like raw green corn, or still more dis-

agreeable. In others no particular odor is noticeable while the plants

are alive, but when the water dries up leaving the algz to die, a

“pig-pen odor” is given off and the water remaining in the pool is

unfit to be used for drinking purposes by animals. This effect is the

result of the breaking down of highly organized compounds of sul-

phur and phosphorus in the presence of the large amount of nitrogen

which these plants contain.

The question of how to keep drinking water pure is creating

much interest at the present time. Water Boards and Boards of

Health in large cities are establishing laboratories where chemists,

botanists and bacteriologists may carry on study and investigation

under the best advantages. Experiments are being made to discover

the best methods of building reservoirs so that light may be shut off

and the water kept free from organic matter, for alge require light

and nutriment for their development.

184 JOSEPHINE E. TILDEN

Thermal Algz

In general all living things are soon killed when subjected to

high temperatures, but certain kinds of algze are able to exist in

water heated almost to the boiling point. Blue-green alge are almost -

the only kinds found in hot waters. In the hot springs and geysers

of Yellowstone Park, those of Iceland, New Zealand, Japan, tropical

Africa, Carlsbad, Arkansas and California, are found these plants.

True hot water alge are those living in water of a temperature rang-

ing from 40° to about 75° C. The upper limit of temperature is not

determined exactly at present.

Calcareous Algz

All green land and water plants—that is, all plants containing

chlorophyll—take in carbon dioxide gas, making the carbon and

some of the oxygen atoms into new compounds for themselves, set-

ting free the rest of the oxygen atoms. The calcium bicarbonate in

water is held in solution by carbonic acid gas. Then anything that

causes the withdrawal of the carbonic acid will cause the lime car-

bonate to be precipitated. Certain alge, in some way that we can not

explain, have this power of withdrawing carbonic acid from the

water in which they live. As the plants grow they become encased

in calcareous sheaths. Perhaps the colony begins to develop on a

piece of shell. The filaments radiate outwards in all directions, each

independent of the others and all precipitating calcium carbonate

tubules. Gradually a pebble of lime is formed looking like an ordi-

nary water-rounded pebble. These algal calcareous pebbles show

both radial and concentric structure. The bluish-green radial lines

are probably formed by growth of filaments. The concentric lines

may represent seasonal or annual periods of growth of plants. These

pebbles have been found in several marl lakes in Michigan and in a

Minnesota lake. Sometimes, also, blue-green alge form calcareous

layers instead of rounded masses or pebbles.

Calcareous hot waters are rare in Yellowstone Park, the one

locality where deposits of travertine have been formed being Mam-

-moth Hot Springs. The travertine deposit here is about two miles

in extent. The thickness of the deposit is variable but is not known

to exceed two hundred and fifty feet. There are about seventy-five

SYNOPSIS OF THE BLUE-GREEN ALGA:-—MYXOPHYCEZ 185

springs. The red, orange and green colors on the overflow slopes are

due to the presence of microscopic alge. This deposit of travertine

is formed chiefly by the activities of alge in withdrawing carbon

dioxide from the water, thus separating carbonate of lime.

The four Geyser Basins of Yellowstone Park are characterized

by the large amount of silica contained in solution by the hot waters,

They are the Norris, Upper, Middle and Lower Basins. Their

waters are alkaline. It is not yet known just how the alge cause

the precipitation of silica from the hot water.

Family Chroococcacez

These plants are unicellular, that is, each individual consists of

a more or less spherical mass of protoplasm, faintly tinged with

bluish-green or purple tints. The wall is usually gelatinous. The

plants exist in an isolated condition or they live together in masses

or colonies imbedded in the gelatinous substance secreted from the

outer layer of their walls. The common method of multiplication is

by simple cell division in one, two or three directions. In Synecho-

coccus division takes place in one direction only and the daughter-

cells remain clinging to one another for a brief period—thus indicat-

ing the origin of the trichome or row of cells. In Merismopedia the

plants form flat, plate-like colonies. The individual plants are

arranged very regularly in the plate, as a result of division taking

place in two directions only. In Aphanothece and Glceocapsa, div-

ision takes place in three directions, so that the colony formed is

approximately spherical.

These plants grow in moist places on rocks, on tree trunks, or

in pools. Many of them are covered by a thick envelope of gelatine

which enables them to endure dryness for a long time.

Merismopedia violacea sometimes forms rose-colored fragile

masses on the bottom of fish tanks or pools.

Microcystis eruginosa is a common pest in many city water

reservoirs during the hot summer months. Ccelosphzrium is also a

common species in city water supplies. The colony is gelatinous,

spherical and hollow. The small, globose, blue-green plants lie in the

peripheral portion immersed in the gelatine. They may be scattered

or lie in pairs or in fours.

186 JOSEPHINE E. TILDEN

Family Oscillatoriaceze

Each trichome is a single plant. Numbers of them form a soft,

filmy dark bluish-green or blackish layer. Some species live in salt

water, but they are particularly common in fresh water stagnant

pools, in ditches, on wet ground, around pumps, cisterns and tanks,

and are often seen as a coating on the inside of wooden watering

troughs.

In structure these plants are the simplest of the filamentous

forms. The trichome is straight or somewhat curved, cylindrical,

composed of a single row of cells placed end to end. In some species

the individual cells are slender and longer than wide, forming a very

thin hair or thread. In other species the cells are broad and short,

and the plant then appears to be made up of a series of discs placed

face to face. The plant is in general cylindrical in shape, but often,

according to the species, it may taper either gradually or abruptly at

each end. The end, or apical cell itself, has a characteristic shape in

the different species. It must always be remembered, when trying to

determine the species of these plants, that they break apart very

easily, thus leaving broken ends which of course do not show the

specific characters of the true apical cell.

In some species a row of granules will be seen on either side of

the common wall.

The family, Oscillatoriacez, takes its name from the fact that

certain alge belonging to it have the power of motion. The move-

ment is a peculiar one and as yet not at all understood. The plant

may move through the water with a gliding motion; it may oscillate

or vibrate; or it may merely bend or wave back and forth at one

extremity, the other end being apparently attached to some object in

the water.

These plants have no heterocysts and no gonidia. They repro-

duce only by means of hormogones. The plant separates into short

segments consisting of a few cells each. These short segments are

motile for a time and are known as hormogones. The cells compos-

ing them begin dividing at once and the hormogone soon attains the

size and appearance of the parent plant.

SYNOPSIS OF THE BLUE-GREEN ALG—MYXOPHYCEZ 187

It is an alga in this family, Trichodesmium erythreum, which

has caused the Red Sea to be so named. It floats in the surface

waters, giving them a red color.

Family Nostocaceze

To this family belong such common plants as Nostoc, Anabzena,

and Cylindrospermum. They are simple in structure, all the cells of

the trichome being somewhat similar in size, shape and appearance. |

The plants are not attached but are isolated or they live in gelatinous

colonies. The sheath surrounding the trichome is either gelatinous

or mucilaginous, and may or may not be visible. The heterocysts

occur at intervals in the trichome, or they may be terminal. The

intercalary heterocysts have two thickened cellulose buttons at the

point of attachment to the adjoining cells. These are wanting in the

terminal heterocysts. When a Nostoc plant forms hormogones, the

sheath softens, allowing the segments of the trichome to escape into

the water. To determine genera and species, the position, form, size

and color of the gonidia must be noted. The character of the wall,

as well as the relation of the gonidium to the heterocyst, must be also

taken into consideration.

Nostoc colonies generally occur as more or less rounded masses,

varying from the size of a pea to that of a plum. Anabzna and

Cylindrospermum form yellowish or bluish-green layers. It is very

important to note that the individual plants in these three genera

resemble each other very closely. Before beginning the study of the

microscopic characters, one must determine whether the colony has

a definite form or whether it occurs merely as a layer without definite

shape.

Some of these plants live inside the tissues of higher plants—in

the roots of Cycas, in the leaves of Azolla, in the fronds and rhizoids

of Liverworts.

Family Scytonemaceze

The filaments (plants) have a base and an apex and are gen-

erally branched. In Scytonema the branches arise in the middle of

the interval comprised between two heterocysts. In Tolypothrix the

branches depart from the sheath immediately below a heterocyst.

The branches may be solitary or in pairs (“twin” branches). The

188 JOSEPHINE E. TILDEN

method of branching, known as “false” branching, is somewhat

peculiar. The growth area occurs midway between two heterocysts

where the cells will be found to be shorter and more crowded. Since

the heterocysts are stationary within the sheath, the increase in the

number of cells and the growth in length of the individual cells create

a strong pressure at this middle point, and it is here that the sheath

will finally give way, allowing the trichome to form a loop outside.

With further growth this ruptures at the bend, so that in general

“twin” false branches are formed. In other cases only one branch

may continue to grow, the other remaining undeveloped. Hormo-

gones, with rounded extremities, may be found within the sheath in

many plants. The sheath is tubular and continuous. It may be

colorless or it may be brownish or yellowish. The heterocysts are

intercalary or basal, solitary or in series. Gonidia have been seen

in some species.

Family Stigonemacez

The most important characteristic of this family is the forma-

tion of true branches. This is effected by a division of the cells in

a direction parallel to that of the main axis. The branch arises by

a development of one of the lateral cells. Thus, a certain lateral

cell divides in the longitudinal direction of the plant body, instead of

transversely. Growth continues in this direction so that there is

formed, projecting at right angles to the main filament, a single row

of cells which is a “true” branch. In species in which the trichome

consists of a single row of cells the heterocyst occupies the place of

an entire cell. When the cells divide longitudinally and there are

several longitudinal rows within the sheath, the heterocyst stands in

the place of one of the lateral peripheral cells. In certain species

having a very strongly colored sheath, they are but slightly apparent.

In general the heterocysts are smaller than the ordinary cells and

have a brighter, more yellowish color. Hormogones are formed at

the ends of branches or in special short branches.

Family Rivulariacez

The plants in this family are distinguished from other forms

of the class by the trichome terminating in a hair. In other words

the plant is wider at the basal portion and tapers towards the apex.

SYNOPSIS OF THE BLUE-GREEN ALGZ—MYXOPHYCE@ 189

The series of cells forming the apical portion, or “hair,” contain little

protoplasm and are not capable of further growth. The cells are

long, slender, straight and colorless. When hormogones are pro-

duced this “hair” falls away. Heterocysts are sometimes wanting.

Sometimes only basal ones exist. Sometimes in the same species

there are both basal and intercalary heterocysts. The plants are

sometimes simple and sometimes they form false branches. Branch-

ing results generally through the growth of the summit of the lower

segment of the trichome, just below an intercalary heterocyst. Rarely

the trichome breaks out in the interval between two heterocysts (most

frequently at the base of old plants of Calothrix). Then the branches

resemble those of Scytonema. Sometimes the branches break 1mme-

diately from the main filament. Sometimes they remain inside the

sheath.

The sheath is tubular, continuous, gelatinous, or membranaceous,

homogeneous or lamellose. The interior layers are often capable of

much swelling. Sometimes particles of lime are deposited upon the

sheath. Then the colony becomes transformed into a stony mass.

The sheaths are colorless or tinted with yellow or brown. Gonidia

are known in but few cases. These are generally submerged plants.

MY XOPHY CEA:

The Myxophycee are typically blue-green, the coloring matter

being a mixture of two pigments, chlorophyll and phycocyanin; pigments

of other colors sometimes present.

The plant body is unicellular or multicellular, sometimes endowed

with a peculiar motion; plants existing usually in-gelatinous masses,

sometimes solitary among other alge.

Reproduction is always asexual, either by simple cell division in one,

two or three directions of space, or by means of hormogones (multicellu-

lar fragments of the plant body, at first motile, afterwards coming to

rest), or by means of non-motile gonidia formed within gonidangia, or by

means of resting gonidia formed from ordinary cells.

Habitat: Plants are found in fresh, brackish or salt water, in hot

springs, in mineral springs, in «rial situations, or as endophytes.

Order I. Coccogonez. Plants unicellular, single or associated in fam-

ilies or colonies which are usually surrounded by a copious gelatinous

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SYNOPSIS OF THE BLUE-GREEN ALG—-MYXOPHYCE 191

integument, rarely forming filaments; reproduction occurs commonly by

the vegetative division of cells, rarely by the formation of non-motile

gonidia from the division of the contents of a gonidangium (mother-cell).

Order II. Hormogonez. Plants multicellular, filamentous, attached to

a substratum or free-floating; filaments simple or branched, usually con-

sisting of one or more rows of cells within a sheath; reproduction occurs

by means of hormogones or resting gonidia. Page 209.

Order I. COCCOGONEZ

Family I. Chroococcacee. Plants showing no difference between basal

and apical regions, solitary or associated in families or colonies; reproduc-

tion by vegetative division of cells in one, two or three directions of space.

Family II]. Chamesiphonacee. Plants often showing a difference be-

tween basal and apical regions, solitary or associated in families or col-

onies, usually epiphytic or attached to shells; reproduction by means of

non-motile gonidia formed by the division of the contents of a mother-

cell (gonidangium). Page 205.

Family I. CHROOCOCCACE/:

I. Plants solitary or associated in small, indefinite families or colonies,

not surrounded by a common (colonial) gelatinous tegument

1 Cells spherical; reproduction by cell division in three directions

Chroococcus

Plate XVII, Fig. 1

Cells spherical; reproduction by cell division in one direction only

Synechocystis

Fig. 2

3. Cells oblong, ellipsoidal or cylindrical; sheath wanting; reproduc-

tion by cell division in one direction only

Synechococcus

| Fig. 3

4 Cells cylindrical or oblong-conical; sheaths thick, hyaline; repro-

duction by cell division in one direction only

Chroothece

Fig. 4

II. Plants associated in families or colonies, surrounded by a common

gelatinous tegument

1 Colonies without definite shape

(1) Individual sheaths usually thick, remaining through many divi-

sions,. sheath of original mother-cell surrounding entire

colony

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PLATE XVIII

SYNOPSIS OF THE BLUE-GREEN ALG#—MYXOPHYCEZ 193

A Cells spherical

a Cells enclosed in a vesicle-like, thick, colorless or colored

sheath, spherical (after division oblong), single or in col-

onies; cell contents blue-green, or of various colors

Glceocapsa

Fig. 5

b Cells surrounded by an elliptical membrane, forming colon-

ies, arranged in short filaments Entophysalis

Fig. 6

ce Cells surrounded by thick sheath, forming spherical colonial

masses; plant mass cushion-like, cartilaginous, incrusted

with lime at base, curled at periphery

Chondrocystis

Figs. 7, 8

B Cells elongate

a Cells cylindrical-oblong, surrounded by a thick, mucous

sheath, solitary or forming small colonies

Gleeothece

Fig. 9

(2) Individual sheaths not distinct; colony surrounded by com-

mon tegument formed of dissolved individual sheaths

A Cells spherical (or angular from mutual pressure); cell divi-

sion in all directions Aphanocapsa

Fig. 10

B Cells oblong; cell division in one direction

Aphanothece

Plate XVIII, Fig. 11

2 Colonies having a definite characteristic shape

(1) Colonies free-floating

A Cells having an indefinite arrangement, forming several layers

a Cells spherical or oblong; colony spherical or oblong, solid

Microcystis

Fig. 12

b Cells spherical; colonies of variable shape, at first solid, be-

coming saccate and clathrate Clathrocystis

Figs. 13, 14

ce Cells pear-shaped or heart-shaped; colony spherical or ellip-

soid, solid Gomphospheria

Fig. 15

B Cells having a definite arrangement, forming a single layer or

cube

a Colonies spherical, hollow

194 JOSEPHINE E. TILDEN

(a) Cells spherical, lying just within the periphery of the

colony Ccelospherium

Fig. 16

(b) Cells spherical or elongate; individual sheaths distinct -

Celosphezriopsis

Fig. 17

b Colonies flat

(a) Cells of some definite or symmetrical shape, quadran-

gular or triangular, solitary or forming colonies

| Tetrapedium

Fig. 18

(b) Cells spherical; colonies rectangular

Merismopedium

Fig. 19

c¢ Colonies cubical, solid; cells spherical or elliptical

Eucapsis

Fig. 20

(2) Colonies adherent to substratum

A Cells spherical or elongate, regularly arranged in radial rows;

colonies cushion-like, hard, leathery, verrucose

Oncobyrsa

Fig. 21

B Cells spherical or oval, irregularly arranged in radial rows;

colonies irregularly lobed, epiphytic Chlorogleea

Fig. 22

Genus CHROOCOCCUS

Plate XVII, Fig. 1

Plants either free-floating or forming a gelatinous or crust-like plant

mass in damp places, in fresh or salt water, or within the tissues of other

plants, occurring as spherical or angular cells, each surrounded by a more

or less definite sheath, solitary or united in twos, fours, eights, etc., but not

held together in- definite colonies by a common gelatinous tegument;

sheaths thin or wide, homogeneous or lamellose, colorless or colored; cell

contents homogeneous or granular, usually of a blue-green color, some-

times violet, olive-green, orange or yellowish; reproduction by succes-

sive division of the cells alternately in three directions of space.

I, Sheaths hyaline, often lamellose; cell contents orange or yellowish.

1 Cells less than 3 mic. in diameter C. rubrapunctus

2 Cells more than 15 mic. in diameter

(1) Plant mass yellowish green; cells 25-50 mic. in diameter

C. macrococcus

(2) Plant mass orange-colored; cells 19-34 mic. in diameter

C. turicensis

SYNOPSIS OF THE BLUE-GREEN ALGH—MYXOPHYCE 195

II. Sheaths hyaline, yellowish or brownish, often lamellose; cell con-

tents blue-green, rarely olive-brown, reddish-green, brownish-

violet or copper-red

1 Cells not embedded in a gelatinous mass, mostly solitary among

other alge

(1) Sheaths thick, distinctly lamellose; cell contents blue-green

A Sheaths colorless; cells 13-25 mic. in diameter

C. turgidus

B Sheaths yellowish or brownish; cells 5.8-11 mic. in diameter

C. schizodermaticus

(2) Sheaths not lamellose

A Cells 5-7 mic. in diameter C. minutus

B Cells 1.7 mic. in diameter C. multicoloratus

C Growing in hot water; cells 1-1.5 mic. in diameter

C. thermophilus

2 Cells embedded in a gelatinous mass, not free-floating

(1) Sheaths lamellose

A Sheaths slightly lamellose; plants 4-8 mic. in diameter

C. varius

B Sheaths lamellose, finally irregularly peeling off; plants 6-11

mic. in diameter C. decorticans

(2) Sheaths not lamellose, sometimes scarcely visible

A Plants 5 mic. in diameter, mostly subquadrate, often triangu-

lar, rarely multiangular; sheaths scarcely perceptible

C. refractus

B Plants 4-7.5, rarely 9 mic. in diameter, spherical

C. helveticus

C Plant mass pale yellowish; sheaths oblong-elliptical; cells

7.5-13 mic. in diameter; cell contents blue-green, yellowish

or orange C. pallidus

D Plant mass green, later becoming black; sheaths distinct,

ellipsoid; cells 2.7-6.6 mic. in diameter; cell contents blue-

green C. cohzrens

E Plant mass blue-green or olive; sheaths scarcely visible;

plants 3-4 mic. in diameter; cell contents blue-green

C. minor

F Plant mass lead-colored or green becoming black; sheaths

thick, mucous; plants 3-8 mic. in diameter; cell contents

blue-green C. membraninus

196 JOSEPHINE E. TILDEN

3 Cells embedded in a gelatinous, free-floating mass

(1) Plants 8-13 mic. in diameter, much crowded; cell contents

green or blue-green C. limneticus

(2) Plants 13 mic. in diameter, usually in groups of two; groups

lying apart from each other; cell contents grayish-purple

C. purpureus

Genus SYNECHOCYSTIS

Plate XVII, Fig. 2

Plants always globose; sheaths none; cell walls thin not diffluent;

cell contents blue-green; reproduction by division of the cells in one direc-

tion only.

Submerged; plants 5-6 mic. in diameter, single or in pairs; cell walls

hyaline, very thin; cell contents pale blue-green. S. aquatilis

Genus SYNECHOCOCCUS

Plate XVII, Fig. 3

Plants oblong, cylindrical or ellipsoidal, usually single, occasionally

forming families of two or four united in a row or chain; sheaths none;

cell walls thin; cell contents blue-green, sometimes yellowish, pinkish or

pale orange; reproduction by division of the cells in one direction only.

I. Cell contents blue-green

1 Cells 7-15 mic. in diameter, 14-26 mic. in length

S. #ruginosus

2 Cells 2 mic. in diameter, 4-6 mic. in length SS. racemosus

3 Growing in hot salt water; cells 3 mic. in diameter, 6 mic. in length

S. curtus

Genus CHROOTHECE

Plate XVII, Fig. 4

Plant mass somewhat gelatinous, dark-yellowish; plants cylindrical

or oblong-conical, with rotund ends, single or in pairs; sheaths wide,

lamellose, hyaline, increasing greatly in thickness at one pole; cell con-

tents distinctly granular, bright blue-green or orange-yellow; reproduc-

tion by division of the cells in one direction only.

I. Plants 18-24 mic. in diameter C. richteriana

II. Plants 1.5 mic. in diameter C. cryptarum

III. Plants 11-12.5 mic. in diameter C. monococca

SYNOPSIS OF THE BLUE-GREEN ALG7Z—MYXOPHYCEZ 197

Genus GLG@OCAPSA

Plate XVII, Fig. 5

Plants spherical (or immediately after division oblong), either single

or a number associated in families; each cell enclosed in a vesicle-like,

strongly thickened, usually distinctly lamellose sheath; sheaths often very

thick, colorless or colored, usually lamellose; lamelle often peeling off;

cell contents blue-green, bluish, steel-blue, reddish, yellowish, etc.; repro-

duction by division of the cells alternately in three directions.

When a cell divides into two daughter-cells, each one secretes a

sheath about itself, the two still being enclosed by the sheath of the

mother-cell. As division goes on, the sheath of the original cell remains

enveloping the entire family, and in fact all the sheaths remain in exist-

ence. Therefore, there will always be one less than twice as many

sheaths as there are cells in the family (in a family of four cells there

will be seven sheaths; in a family of sixteen cells there will be thirty-one

sheaths). Later generations of cells are smaller than the first ones.

I. Sheaths colorless

1 Sheaths lamellose

(1) Sheaths wide

A Plant mass steel blue, green, olive or dull yellow; plants 7-8

mic. in diameter; sheaths very wide, indistinctly lamel-

lose; cells 3-5 mic. in diameter G. granosa

B Plant mass dull green or olive; plants 3-4.5 mic. in diameter;

sheaths very thick, with numerous concentric lamellz

G. polydermatica

C Plant mass green; plants 7-15 mic. in diameter; sheaths very

thick, more or less distinctly lamellose; cells 2.2-3.4 mic.

in diameter G. fenestralis

D Plant mass somewhat olivaceous; plants 6-17 mic. in dia-

meter; sheaths thick; cells 3.7-6 mic. in diameter

G. arenaria

(2) Sheaths narrow

A Plant mass pale yellow becoming greenish; growing in hot

water; plants 19-39 mic. in diameter; cells 3-6 mic. in dia-

meter . G. montana

B Plant mass mucilaginous, dull green or gray becoming black-

ish, or red becoming brownish; plants 7-11 mic. in dia-

meter; cells 3-4.5 mic. in diameter G. quaternata

C Plant mass a calcareous crust, light gray or green; plants 6-9

mic. in diameter | G. calcarea

198 JOSEPHINE E. TILDEN

D Plant mass gelatinous, brownish, growing on Zostera; sheaths

numerous, distinct; cells 9-11 mic. in diameter, 19-26 mic. in

length G. zostericola

2 Sheaths sometimes lamellose

(1) Plant mass blue-green or greenish; sheaths not distinctly

lamellose

A Free-floating; cells .75-2.8 mic. in diameter

G. punctata

B On wet rocks; plants 48 mic. in diameter; cells 2-3 mic. in

diameter G. zruginosa

(2) Plant mass olive or green; plants 62-10 mic. in diameter;

sheaths narrow, lamellose when old; cells 2.5 mic. in dia-

meter G. gelatinosa

(3) Plant mass dull olive; plants 7-11 mic. in diameter; sheaths

thick, not at all or scarcely lamellose; cells 3-6 mic. in diam-

eter G. conglomerata

3 Sheaths not lamellose

(1) Plant mass fiesh-colored to yellowish; plants 2.5-5.5 mic. in

diameter; cell contents flesh-colored to honey-colored

G. mellea

(2) Plant mass black; plants 9-14 mic. in diameter: cells 3.5-4.5

mic. in diameter; cell contents pale blue-green

G. atrata

II. Sheaths yellowish or brownish

1 Sheaths lamellose

(1) Plant mass dull olive to brownish-green: sheaths colorless

or yellowish G. muralis

(2) Plant mass grayish-brown to black: sheaths very thick, yel-

lowish or orange, becoming darker G. rupestris

2 Sheaths sometimes lamellose

(1) Plants 4.5-5.5 mic. in diameter; sheaths usually not lamellose:

cells 1.5-2 mic. in diameter G. fusco-lutea

(2) Plants 12 mic. in diameter; sheaths homogeneous or lamel-

lose; cells 3-4.5 mic. in diameter G. sparsa

(3) Colonies subglobose: sheaths somewhat lamellose; cells 9-15

mic. in diameter G. gigas

3 Sheaths not lamellose; plant mass olive-green; plants 5-8 mic. in

diameter G. crepidinum

SYNOPSIS OF THE BLUE-GREEN ALG#—MYXOPHYCEZ 199

III. Sheaths violet, purple or red

1 Sheaths lamellose

(1) Plant mass purple, sometimes becoming black

A Sheaths deep purple or copper-brown; plants 6-12 mic. in

diameter G. magma

B Sheaths violet or reddish-purple; plants 7.5-12 mic. in diame-

ter; cells 2-4.5 mic. in diameter G. janthina

C Sheaths very thick, opaque, intensely lamellose; plants 10-17

mic. in diameter; cells 4-7 mic. in diameter

G. ralfsiana

2 Sheaths sometimes lamellose; plant mass colorless or dark purple,

growing in hot water; plants 6-7.8 mic. in diameter; cells 1-2.6

mic. in diameter G. thermalis

3 Sheaths not lamellose

(1) Plant mass violet becoming gray or black

A Plants 4-8 mic. in diameter; sheaths violet, thick, often

opaque; cells 1.8-2.5 mic. in diameter G. ambigua

B Plants 10-17 mic. in diameter; sheaths violet or rose-colored;

cells 3.5 mic. in diameter G. violacea

(2) Plant mass reddish-orange, dark red or black

A Plants 11-24 mic. in diameter; sheaths very thick, soon peeling

off G. dubia

B Sheaths intensely blood-red, very wide; cells 3.5-9 mic. in

diameter G. sanguinea

Genus ENTOPHYSALIS

Plate XVII, Fig. 6

Plant mass globose, cartilaginous, including numerous, more or less

confluent small families of cells; cells spherical, each surrounded by an

elliptical sheath, associated in families.

I. Plant mass crustaceous; cells 2-5 mic. in diameter

E. granulosa

II. Plant mass mucous; cells 4-6 mic. in diameter

E. magnoliz

Genus CHONDROCYSTIS

Plate XVII, Figs. 7, 8

Plant mass cushion-shaped, widely expanded, up to 35 cm. high,

cartilaginous, soft, fragile, encrusted with lime at the base, curled up at

periphery; families consisting of spherical masses of cells lying free, the

membranes of which seem to be thickened into one layer.

200 JOSEPHINE E. TILDEN

Plant mass rose-colored to red, thick; cells somewhat spherical

or elongate, 2 mic. in diameter, 3-5 mic. in length; sheath thick,

C. schauinslandii

Genus GLG@OTHECE

Plate XVII, Fig. 9

Colonies embedded in a common gelatinous tegument; cells cylindri-

cai-oblong, rounded at the ends, each surrounded by a wide mucous

homogeneous or lamellose sheath; reproduction by transverse division

of the cells in one direction only.

I. Individual sheaths colorless

1 Cells .8-2.5 mic. in diameter, 10.5-18 mic. in length

G. linearis

2 Cells 1.6-3 mic. in diameter, 2.2-7.5 mic. in length

G. confluens

3 Plants 8-12 mic. in diameter, 12-36 mic. in length; cells 4-5.5 mic.

in diameter, 6-15 mic. in length G. rupestris

4 Cells 4-5 mic. in diameter, 6-10 mic. in length

G. membranacea

5 Cells 2.5-2.7 mic. in diameter, 4.8-5.7 mic. in length, somewhat

crescent-shaped with acute apices G. lunata

II. Individual sheaths partly or entirely colored

1 Plant mass usually free-floating

(1) Sheaths colorless at margins; cells 3-4 mic. in diameter

G. magna

(2) Sheaths usually brownish or yellowish; cells 4-5.5 mic. in

diameter, 6-11 mic. in length G. fuscolutea

Genus APHANACAPSA

Plate XVII, Fig. 10

Plant mass more or less expanded, colorless or blue-green, yellow

or brown; plants spherical or angular from mutual pressure, single or in

pairs; individual sheaths thick, very soft, colorless, not distinct, confluent

into a mucous, amorphous, homogeneous colonial tegument; tegument

colorless or tinted brown or blue-green; reproduction by successive divi-

sion of the cells alternately in three directions.

I. Plant mass colorless

1 Cells 1.5-2 mic. in diameter _ A, elachista

2 Cells 10-16 mic. in diameter A. zanardinii

SYNOPSIS OF THE BLUE-GREEN ALG—MYXOPHYCEZ 201

II. Plant mass green or blue-green

1 Plant mass globose, gelatinous, dirty green; cells 3.2-5.6 mic. in

diameter A. grevillei

2 Plant mass hemispherical, gelatinous, blue-green; cells 5-6 mic.

in diameter A, rivularis

3 Plant mass amorphous, gelatinous, dirty green or olive

A. virescens

III. Plant mass brown

Cells 4.5-5.5 mic. in diameter A. brunnea

Genus APHANOTHECE

Plate XVIII, Fig. 11

Plant mass more or less expanded, somewhat spherical or without

definite shape; individual sheaths thick, not distinct, confluent into a

mucous, amorphous, homogeneous colonial tegument; cells oblong; re-

production by division of the cells in one direction only.

I. Plant mass without definite shape

1 Cells 1-2 mic. in diameter A. saxicola

2 Cells more than 2 mic. in diameter

(1) Plant mass dirty green or olive brown; cells 2.5-3 mic. in

diameter A. conferta

(2) Growing in very salt water; cells 5 mic. in diameter, hardly

longer than broad A. utahensis

(3) Cells one to three times as long as broad

A Plant mass colorless; cells 4-4.5 mic. in diameter

A. microscopica

B_ Plant mass colored

a Plant mass blue-green, olive or yellowish-brown; cells 2-3.5

mic. in diameter A. castagnei

b Plant mass pale blue-green; cells 3-8 mic. in diameter

A. pallida

c Plant mass pale yellowish-green or olive; cells 2-3 mic. in

diameter A. microspora

d Plant mass yellowish-brown or olive; cells 4-4.5 mic. in

diameter, irregularly scattered A, negelii

II. Plant mass more or less spherical

1 Plant mass pale blue-green; cells 3-5 mic. in diameter

A. stagnina

2 Plant mass bright or dark emerald green; cells 5-6.5 mic. in diam-

eter A. prasina

202 JOSEPHINE E. TILDEN

Genus MICROCYSTIS

Plate XVIII, Fig. 12

Colonies spherical or somewhat spherical, solid, finally becoming

hollow and lobed, single or associated in clusters, containing large num-

bers of cells, surrounded by a colorless, gelatinous tegument; cells spheri-

cal, oval or elliptical; cell contents green or blue-green, often showing

vacuoles; reproduction by cell division in three directions.

I. Cells spherical

1 Colonies more or less spherical, usually containing several daugh-

. ter colonies each surrounded by its own tegument; cells 2-4 mic.

in diameter M. ichthyoblabe

2 Plant mass dull yellowish becoming olive; colonies 30-70 mic. in

diameter; cells 2.2-4 mic. in diameter M. donnellii

3 Colonies spherical, flattened, orbicular, lens-shaped, sometimes

confluent, surrounded by a thick, lamellose common tegument;

cells 3-4 mic. in diameter M. marginata

4 Colonies more or less spherical or oblong, with an indistinctly

limited tegument, pale or yellowish-green; cells 4-6.5 mic. in

diameter M. flos-aquz

II. Cells oval or oblong, sometimes almost spherical

1 Colonies spherical, oblong or flattened, sometimes containing sev-

eral daughter colonies each surrounded by its own tegument;

cells 1-1.5 mic. in diameter, 3-5 mic. in length, oblong

M. elabens

2 Plant mass pulverulent, bright glaucous or whitish blue-green;

colonies spherical or oblong; cells 2-3 mic. in diameter, some-

what spherical or oval M. pulverea

3 Plant mass irregular, firm, gelatinous, pink, brown or green, grow-

ing in very salt water; cells 2.5-4 mic. in diameter, 6-7 mic. in

length, oblong or elliptical M. packardii

4 Plant mass mucous, floccose, amorphous, sky-blue; colonies some-

what spherical, distinctly limited; cells somewhat spherical or

ellipsoid (size unknown) M. piscinalis

5 Colonies irregular in shape, with an indistinctly limited tegument;

cells 5-5.5 mic. in diameter, spherical or oval

M. pallida

Genus CLATHROCYSTIS

Plate XVIII, Figs. 13, 14

Colonies of variable shape, at first solid soon becoming saccate and

clathrate, (“fragments of the broken fronds occurring in irregularly lobed

SYNOPSIS OF THE BLUE-GREEN ALG#—MYXOPHYCEH 203

forms”), surrounded by a colorless, gelatinous, indistinctly limited integu-

ment; cells spherical, numerous.

I. Cells 3-4 mic. in diameter, spherical C, zruginosa

II. Cells 6-9 mic. in diameter, spherical or oval C. robusta

Genus GOMPHOSPHAERIA

Plate XVIII, Fig. 15

Colonies spherical or ellipsoid, mucous, solid, free-swimming; tegu-

ment colorless or yellowish, usually thick, soon diffluent; cells pear-

shaped or heart-shaped, rarely somewhat spherical, grouped in pairs, few

in numbers, disposed chiefly towards the periphery of the tegument; cell

contents often granular, bluish or greenish; reproduction by cell division

alternately in three directions.

I. Cells 4-5 mic. in diameter, 8-12 mic. in length G. aponina

II. Cells 3.2-4 mic. in diameter, spherical; cell contents pinkish or

brownish G. rosea

Genus CHLOSPHZRIUM

Plate XVIII, Fig. 16

Colonies spherical, mucous, hollow, free-swimming, containing many

small cells; tegument mucous, soon confluent; cells globose, elliptical or

ovoid, arranged just within the periphery of the tegument; cell contents

granular, with gas vacuoles; reproduction by cell division, at first in one

direction, afterwards alternately in three directions.

I. Colonies 30-90 mic. in diameter; cells 2-5 mic. in diameter

C. kuetzingianum

II. Colonies about 150 mic. in diameter; cells 5-7 mic. in diameter

C. dubium

Genus CHLOSPHZRIOPSIS

Plate XVIII, Fig. 17

Colonies spherical, gelatinous, hollow; families clustered; cells

spherical or elongate, arranged in a single peripheral layer; reproduction

by cell division.

Colonies 30-500 mic. in diameter, spherical, gelatinous; cells 6 mic.

in diameter, 6-9 mic. in length, spherical or elongate.

C. halophila

Genus TETRAPEDIUM

Plate XVIII, Fig. 18

Cells solitary or occurring in families of from 2-16 each, compressed,

quadrangular or triangular, equilateral, becoming subdivided into quad-

204 JOSEPHINE E. TILDEN

rate or wedge-shaped segments or rounded lobes, either by deep verti-

cal or oblique incisions or by wide angular or rounded sinuses; cell con-

tents blue-green; reproduction by cell division. (Single cells break apart

by the incisions into four daughter-cells each, the daughter-cells after

division forming separate individuals. The direction of the incisions is

either perpendicular to the lateral margin-or bisects the angles.)

Cells 3.6x7.2 mic., triangular, with conclave sides and somewhat

rotund angles, elliptical in side view; cell contents homogeneous, pale

blue-green. T. trigonum

Genus MERISMOPEDIUM

Plate XVIII, Fig. 19

Colonies flat, rectangular, free-floating; tegument somewhat thick,

confluent; cells spherical, before division oblong, arranged in a rectilinear

series in a single layer; cell contents usually without gas vacuoles, blue-

green, rarely violet, rose-pink or red; reproduction by division of the

cells in two directions.

I. Cells 5-7 mic. in diameter, 6-9 mic. in length

1 Colonies 30 mic. in diameter; cells 5 mic. in diameter, somewhat

spherical; cell contents blue-green or violet

M. zrugineum

2 Cells spherical or oblong M. elegans

3 Cells oval M. novum

II. Cells 3-6 mic. in diameter

1 Colonies 45-150 mic. in diameter; cells spherical or oval

M. glaucum

2 Colonies large, more or less convolute; cells spherical or oblong

M. convolutum

III. Cells 1.3-2 mic. in diameter, somewhat spherical

M. tenuissimum

Genus EUCAPSIS

Plate XVIII, Fig. 20

Colonies cubical, usually consisting of 32-128 cells, but ranging from

8-512 cells, free-floating; tegument uniform, colorless, gelatinous; cells

spherical, sometimes elliptical or flattened by mutual pressure, forming

cubical families; cell contents finely granular, blue-green; reproduction by

cell division in three planes.

Colonies 30-80 mic. in diameter, usually containing 32-128 cells, free-

floating; cells 6-7 mic. in diameter. E. alpina

Oe ee

SYNOPSIS OF THE BLUE-GREEN ALGAt—MYXOPHYCEZz 205

Genus ONCOBYRSA

Plate XVIII, Fig. 21

Colonies cushion-like, hard, leathery, adherent; sheaths thick, gelat-

inous, confluent; cells spherical or elongated, usually regularly arranged

in radial rows; cell contents blue-green or violet

I. Cells pale blue-green, sometimes violet O. rivularis

II. Cells bright blue-green O. cesatiana

Genus CHLOROGLG@A

Plate XVIII, Fig. 22

Colonies irregularly lobed; tegument thin, not lamellose; cells spher-

ical or oval, arranged in radiating series; reproduction by cell division in

one direction.

Colonies disc-shaped, epiphytic, greenish; cells 1-1.5 mic. in diam-

eter, 2 mic. in length; ellipsoid, after division somewhat spherical.

C. tuberculosa

Family II. CHAMAZESIPHONACEZ:

Plants often showing a difference between basal and apical regions,

solitary or associated in families or colonies, usually epiphytic or attached

to shells; reproduction by cell division, by division of filaments into frag-

ments, or by means of non-motile gonidia formed by the division of the

contents of a mother-cell or gonidangium.

I. Reproduction by cell division and by gonidia; cells usually united in

colonies

1 Colonies somewhat spherical or hemispherical, usually consisting

of several layers of cells Pleurocapsa

Plate XIX, Fig. 23,

2 Colonies disc-shaped, usually consisting of a single layer of cells

Xenococcus

Fig. 25

3 Colonies forming branched filaments Hyella

Fig. 26

II. Reproduction by gonidia only

1 Gonidia formed by simultaneous division of the entire contents of

gonidangium Dermocarpa

Fig. 27

2 Plants not usually united in colonies; gonidia formed by successive

constrictions of apical portion of contents of gonidangium

Chamesiphon

Figs. 28, 29

SYNOPSIS OF THE BLUE-GREEN ALG—MYXOPHYCE 207

Genus PLEUROCAPSA

Plate XIX, Figs. 23, 24

Colonies usually crustaceous, made up of vegetative cells and gonid-

angia; plants united in short filaments, parallel or scarcely distinct, radiat-

ing, often dichotomously divided; cells spherical or angular, rarely oval or

polyhedral; cell contents blue-green, olive, yellowish or violet; gonidangia

furnished with thick sheaths, producing numerous, spherical gonidia;

reproduction by cell division in three directions, by division of filaments

into fragments, and by gonidia formed by division of the contents of a

gonidangium.

I. Cells arranged in straight rows; growing in fresh water

Pl. concharum

II. Cells not arranged in straight rows

1 Growing in hot water; cells 4-6 mic. in diameter

Pl. caldaria

2 Growing in salt water

(1) Cells 5-20 mic. in diameter; cell contents golden yellow, fawn-

colored or dull violet Pl. fuliginosa

(2) Cells 10-13 mic. in diameter; cell contents violet

Pl. amethystea

(3) Cells up to 15 mic. in diameter; cell contents dull blue or slate

color Pl, crepidinum

Genus XENOCOCCUS

Plate XIX, Fig. 25

Colonies disc-shaped or crustaceous, attached; cells somewhat spher-

ical, or angular with rounded apices, crowded, forming a parenchymatous,

one-celled layer, later several cells in thickness; tegument colorless or yel-

lowish; cell contents homogeneous, blue-green or violet; reproduction by

cell division in three directions or by means of gonidia developed in large

peripheral cells; gonidia usually spherical, sometimes 32 developed in a

gonidangium.

I. Colonies disc-shaped, composed of one layer of cells; tegument sur-

rounding base of cells; cells 3-4 mic. in diameter, 5.5-7 mic. long,

pear-shaped X. laysanensis

II. Colonies spherical, solitary or confluent and completely surrounding

the filaments of the host; cells 4-9 mic. in diameter, spherical or

flattened X. schousbeei

IlI. Colonies irregularly expanded, one or several layers in thickness;

cells 4-6 mic. in diameter, 4-9 mic. in length

X. kerneri

208 _ JOSEPHINE E. TILDEN

Genus HYELLA

Plate XIX, Fig. 26

Colonies radiately expanded, orbicular, composed of two kinds of

filaments; primary filaments horizontal, tangled, twisted, finally becom-

ing a very densely woven felty mass; secondary filaments vertical, devel-

oped from primary; branching true; tegument septate, thicker at base of

filament, narrower above; cells disconnected, not joined in chains, lower

ones short, sometimes divided longitudinally, upper ones longer; repro-

duction by means of vegetative cells set free from tegument and by means

of gonidia formed in gonidangia by successive division of contents.

I. Colonies yellowish or olive, at first forming minute patches or dots,

later becoming membranaceous or cushion-shaped; erect fila-

ments usually parallel; vegetative cells usually 5-6, sometimes up

to 10 mic. in diameter. H. cespitosa

II. Colonies immersed in substance of shell, brownish-gray or bright

blue; vegetative cells 5-10 mic. in diameter H. fontana

Genus DERMOCARPA

Plate XIX, Fig. 27

Colonies usually epiphytic, forming a somewhat indefinite layer;

cells spherical, egg-shaped, pear-shaped, oval or oblong, solitary or united

in a layer; cell contents usually blue-green or violet; reproduction by

means of gonidia formed by simultaneous division of contents of the

gonidangium; gonidangia oval or elongate, dissolving at apex to allow

the escape of the gonidia.

I. Cells somewhat oval or oblong, not contracted at base to form a

stalk

1 Cell contents blue-green, green, olive or brown

D. prasina

2 Cell contents rose-colored or violet

(1) Cells 4-5 mic. in diameter D. rosea

(2) Cells 8-28 mic. in diameter D. violacea

II. Cells contracted at base to form a stalk

1 Colonies dark violet-brown; cells 18-25 mic. in diameter, 40-60

mic. in length D. fucicola

2 Colonies irregularly outlined; cells 8.5-11 mic. in diameter, 16.5-33.5

mic. in length D, smaragdinus

3 Colonies minute; cells 9.5-17 mic. in diameter, 13-25 mic. in length

D. olivaceus

4 Cells 18-24 mic. in diameter, 17-24 mic. in length

D. leibleiniz

' var. pelagica

SYNOPSIS OF THE BLUE-GREEN ALG4—MYXOPHYCEZ 209

Genus CHAMASIPHON

Plate XIX, Figs. 28,29

Plants epiphytic, erect, cylindrical, somewhat filiform, club-shaped

or pear-shaped, attached at base, widening upwards to free apex, solitary

or aggregated; sheaths present; cell walls very thin; cell contents homo-

geneous, blue-green, violet or yellow; reproduction by one-celled, non-

motile gonidia which are successively cut off from the upper portion of

the contents of the gonidangium, gradually escaping from the open apex.

I. Gonidangia usually 1-2 celled Ch, incrustans

II. Gonidangia many-celled Ch. curvatus

Order II. HORMOGONEZ

Plants multicellular, filamentous, attached to a substratum or free-

floating; filaments simple or branched, usually consisting of one or more

rows of cells within a sheath; reproduction occurs by means of hormo-

gones or resting gonidia.

Family I. Oscillatoriacez. Filaments frequently branched, containing

one or more trichomes; sheaths variable, more or less gelatinous; tri-

chomes consisting of a simple row of cells uniform along their entire

length, except for the apical cells which sometimes taper more or less;

heterocysts absent; reproduction by means of vegetative division and

hormogones.

Family II. Nostocacez. Sheaths very delicate, mostly confluent, usually

not visible; trichomes usually twisting and entangled, consisting of a sin-

gle row of uniform cells, with heterocysts; reproduction by means of

vegetative division, hormogones and gonidia. Page 231.

Family III. Scytonemacez. Filaments with a false branch system;

sheaths firm and tubular; trichomes consisting of a single row of cells,

but not of uniform thickness, with heterocysts; reproduction by means of

vegetative division, hormogones and gonidia. Page 241.

Family 1V. Stigonemacee. Filaments frequently branched; sheaths

thick, firm, often irregular; trichomes consisting of one or several rows

of cells, with heterocysts; reproduction by means of vegetative division,

hormogones and gonidia. Page 247.

Family V. Rivulariacee. Filaments tapering from the base to the

apex, ending in a multicellular, colorless hair; heterocysts usually present,

basal; reproduction by means of vegetative division, hormogones and

gonidia. Page 251.

Family I. OSCILLATORIACEZ

Filaments frequently branched, containing one or more trichomes;

sheaths variable, more or less gelatinous; trichomes consisting of a simple

SYNOPSIS OF THE BLUE-GREEN ALGEH—MYXOPHYCEZ 211

row of cells uniform along their entire length; except for the apical cells

which sometimes taper more or less; heterocysts absent; reproduction by

means of vegetative division and hormogones.

I. Sheaths not present

1 Trichomes straight or nearly so, never forming a regular spiral

(1) Trichomes cylindrical, usually without sheaths, free; apex of

trichome straight or curved Oscillatoria

Plate XX, Figs. 30,

31, 32

(2) Trichomes cylindrical, without sheaths, united in free-swim-

ming scale-like masses Trichodesmium

Fig. 33

2 Trichomes forming a regular, more or less lax spiral

(1) Trichomes multicellular Arthrospira

Fig. 34

(2) Trichomes unicellular Spirulina

Fig. 35

II. Sheaths present

1 Filaments simple or branched; sheaths cylindrical, firm; trichomes

single within the sheath; apex of trichome straight

(1) Filaments simple, more or less agglutinated by their mucous

sheaths Phormidium

f Fig. 36

(2) Filaments simple, free, free-floating or forming a matted mass

Lyngbya

Fig. 37

(3) Filaments often branched, forming erect tufts; false branches.

solitary Symploca

Fig. 38

(4) Filaments simple; sheaths usually purple or flesh-colored;

apical cell not capitate Porphyrosiphon

Fig. 39

2 Filaments frequently branched; sheaths firm, lamellose, trans-

parent or colored; trichomes several within the sheath

(1) Sheaths more or less mucous, colorless, diffluent; trichomes

few within the sheath; apex of trichome capitate

Hydrocoleus

Figs. 40, 41

(2) Filaments prostrate, woven into a solid membranaceous mass,

often slightly branched; sheaths solid, always thin, color-

less; plants terrestrial or aquatic Hypheothrix

Plate XXI, Fig. 42

(3) Filaments prostrate at the base, above forming erect tufts;

sheaths solid, transparent; plants terrestrial

Symplocastrum

Figs, 43, 44, 45

212 JOSEPHINE E. TILDEN

(4) Filaments tufted, often much branched; sheaths transparent or

scarcely colored; plants low, aquatic Inactis

Fig. 46

(5) Filaments branched; sheaths solid, closed at the apex, of vari- »

ous colors; trichomes densely aggregated within the sheath

Schizothrix

Fig. 47

(6) Sheaths wide, transparent or yellowish brown; trichomes

very few within the sheath, very loosely aggregated

Dasygloea

Fig. 48

(7) Sheaths mucous, not lamellose, always transparent; trichomes

many within the sheath Microcoleus

Fig. 49

3 Colonies somewhat spherical, elliptical or spindle-shaped; fila-

ments solitary or aggregated in colonies; sheaths thick, gelatin-

ous

(1) Sheaths very thick; trichomes usually single or in scattered

fragments Catagnymene

Fig. 50

(2) Colonies somewhat spherical; sheaths thick; trichomes curved,

radiating Haliarachne

Genus OSCILLATORIA

Plate XX, Figs. 30, 31, 32

Trichomes cylindrical, free, motile, without a sheath or rarely en-

closed in a very thin, fragile, mucous sheath, often constricted at the

joints; apex of trichome straight, curved, or more or less regularly

spiralled, often tapering; outer wall of apical cell often thickened, form-

ing a calyptra.

I. Plants living in fresh water, floating; apex of trichome constantly

straight, gradually tapering, obtuse, finally capitate; cells some-

what quadrate or shorter than the diameter, never very short

1 Plant mass purple; trichomes 2.2-5 mic. in diameter; cells some-

what quadrate or longer than the diameter

O. prolifica

2 Plant mass light blue-green; trichomes 4-6 mic. in diameter; cells

somewhat quadrate or twice as short as the diameter

O. agardhii

II. Plants living in fresh water, sometimes in hot water; trichomes

large or very large; apex of trichome straight, curved or spiral,

not at all or briefly tapering, obtuse; cells very short

SYNOPSIS OF THE BLUE-GREEN ALG4—MYXOPHYCE2 213

1 Transverse walls never granulated

(1) Trichomes 16-60 mic. in diameter; apex of trichome slightly

tapering, somewhat capitate, hooked O. princeps

(2) Trichomes 12-15 mic. in diameter; apex of trichome tapering,

capitate, hooked or loosely terebriform

O. proboscidea

2 Transverse walls frequently granulated

(1) Apex of trichome straight

A Trichomes 10-20 mic. in diameter, constricted at joints; apex

of trichome very briefly tapering, somewhat capitate

O. sancta

‘\

B. Trichomes 11-20 mic. in diameter, not constricted at joints;

apex of trichome neither tapering nor capitate

O. limosa

(2) Apex of trichome spiral, rarely hooked

A. Trichomes 10-17 mic. in diameter, not constricted at joints;

apex of trichome not capitate O. curviceps

B Trichomes 18-23 mic. in diameter; apex of trichome slightly

tapering, obtusely rounded, usually straight

O. major

C Trichomes 9-11 mic. in diameter, slightly constricted at joints,

here and there interrupted by inflated, refringent cells;

apical cell not capitate O. ornata

D Trichomes 6-8 mic. in diameter, not constricted at joints,

here and there interrupted by inflated, refringent cells;

apical cell capitate O. anguina

III. Plants living in salt water; trichomes always constricted at joints,

rarely straight or spiral throughout; apex of trichome scarcely

tapering, very gradually curved, obtuse

1 Trichomes twisted into a regular spiral O. bonnemaisonii

2 Trichomes not spiral, gradually curved in apical portion, rarely

straight

(1) Plant mass dull red; trichomes 16-24 mic. in diameter

O. miniata

(2) Plant mass olive green; trichomes 17-29 mic. in diameter

O. margaritifera

(3) Plant mass dark olive green; trichomes 7-11 mic. in diameter,

straight, fragile QO. nigro-viridis

(4) Plant mass thin, fragile; trichomes 9.6-11.9 mic. in diameter,

sometimes spirally coiled, sometimes curved or even nearly

straight O. capitata

214 JOSEPHINE E. TILDEN

(5) Plants epiphytic; trichomes 6-10 mic. in diameter, flexuous,

flexible O. coralline

IV. Plants living in fresh water, sometimes in hot water; trichomes

straight or curved, not tapering at the apices .

1 Trichomes 8.5 mic. in diameter, straight or slightly flexuous

O. nigra

2 Trichomes 4-10 mic. in diameter, usually slightly constricted at the

joints, often curved at the apices; transverse walls usually fur-

nished with two rows of granules O. tenuis

3 Trichomes 2-3 mic. in diameter, not constricted at joints, curved

at the apices; transverse walls commonly marked by two

protoplasmic granules O. amphibia

4 Trichomes 1-1.5 mic. in diameter, straight or rolled in a circinate

manner O. subtilissima

5 Trichomes 2.3-4 mic. in diameter, curved, very much constricted at

joints; transverse walls pellucid, not granulated

O. geminata

6 Trichomes 2.5 mic. in diameter, especially constricted at joints;

transverse walls pellucid O. minnesotensis

7 Trichomes 3.5-4 mic. in diameter, not constricted at joints;

transverse walls pellucid, not granulated O. chlorina

8 Trichomes .6 mic. in diameter, flexible, elongate, tangled, not con-

stricted at joints O. angustissima

V. Plants living in fresh water, hot water, rarely in salt water; tri-

chomes tapering, more or less pointed, hooked or flexuous, not

entirely spiralled (except O. chalybea); cells longer or shorter

than the diameter, never very short

1 Apical cell capitate

(1) Trichomes 2-3 mic. in diameter; cells longer than their diam-

eter O. splendida

(2) Trichomes 2.5-5 mic. in diameter; cells somewhat quadrate

O. amena

2 Apical cell not capitate

(1) Piants living in salt water

A Trichomes 4.7-6.5 mic. in diameter, flexible, undulating; apex

of trichome very gradually tapering, very flexuous

O. subuliformis

B. Trichomes 4 mic. in diameter, somewhat flexuous, sometimes

coiled in a regular circle, very much constricted at joints;

apex of trichome tapering, slightly curved, obtuse

O. salinarum

SYNOPSIS OF THE BLUE-GREEN ALG2—MYXOPHYCE 215

C Trichomes 3-5 mic. in diameter, fragile, straight; apex of tri-

chome briefly tapering, hooked or undulating ,

O. lete-virens

(2) Plants living in fresh water, often in hot water, rarely in

brackish water

A Trichomes 3-5 mic. in diameter; apex of trichome briefly

tapering, very sharply pointed, hooked; cells usually longer

than their diameter O. acuminata

B Trichomes 3-4 mic. in diameter; apex of trichome briefly

tapering, very sharply pointed, hooked; cells usually short-

er than their diameter O. animalis

C. Trichomes 4-4.5 mic. in diameter, straight, entangled; trans-

verse walls granulated; cell contents violet or sky-blue

O. violacea

D Trichomes 4-6.5 mic. in diameter, here and there interrupted

by inflated refringent cells; apex of trichome briefly taper-

ing, hooked or flexuous; cells three times shorter than

their diameter O. brevis

E Trichomes 4-7 mic. in diameter; apex of trichome obtuse,

straight, rarely slightly curved O. cruenta

F Trichomes 4-6 mic. in diameter, slightly constricted at joints;

apex of trichome briefly and somewhat obtusely tapering,

hooked; cells quadrate or one-half as long as wide

O. formosa

G Trichomes 2.5-4 mic. in diameter, constricted at joints; apex

of trichome very gradually tapering, hooked or undulating;

cells quadrate or longer than the diameter

O. numidica

H Trichomes 5.5-8 mic. in diameter, slightly constricted at

joints; apex of trichome very gradually tapering, hooked or

undulating; cells quadrate or longer than the diameter,

very long near the apex O. cortiana

I Trichomes 5.5-9 mic. in diameter, constricted at joints; apex

of trichome very gradually tapering, undulating and finally

hooked; apical cell obtuse; cells shorter than their diameter

O. okeni

J Trichomes 8-13 mic. in diameter, scarcely constricted at

joints, sometimes twisted in loose spirals; apex of trichome

briefly or gradually tapering and hooked; apical cell ob-

tuse; cells shorter than their diameter

O. chalybea

216 JOSEPHINE E. TILDEN

K Trichomes 8-10 mic. in diameter, straight, somewhat con-

stricted at joints; apex of trichome often slightly tapering,

obtuse, straight or curved O. subsalsa

L Trichomes 15.5-18.5 mic. in diameter, straight; apex of tri-

chome usually curved, somewhat tapering, obtuse-truncate

O. percursa

VI. Plants living in fresh water, sometimes in hot water; trichomes

regularly terebriform in apical portion or forming a spiral

throughout their entire length, more or less tapering in the

apical portion

1 Trichomes 6-8 mic. in diameter, forming a lax and regular spiral

through their entire length, or straight and hooked at the apex;

apical cell pointed, not capitate O. boryana

2 Trichomes 4-6.5 mic. in diameter, flexuous, straight below, loosely

spiralled and terebriform above; apical cell obtuse, not capitate

O. terebriformis

Genus TRICHODESMIUM

Plate XX, Fig. 33

Plants forming scale-like, disconnected, free-floating colonies quick-

ly dissolving into mucous; trichomes cylindrical, without sheaths; apex of

trichome straight, tapering, slightly capitate; apical cell truncate-conical,

furnished with a convex calyptra.

Floating in great abundance in the ocean, especially in equatorial

regions.

I. Trichomes straight T. erythreum

II. Trichomes flexuous or spirally twisted

1 Colonies up to 6 mm. in length; trichomes 7-16 mic. in diameter,

not constricted at joints, those in center of colony having the

form of twisted ropes, free at the ends T. thiebautii

2 Colonies spirally twisted, light yellow; trichomes 16-25 mic. in

diameter, twisted together into cords T. contortum

Genus ARTHROSPIRA

Plate XX, Fig. 34

Trichomes multicellular, cylindrical, without a sheath, forming a

very regular, more or less loose spiral; apex of trichome sometimes taper-

ing; apical cell rotund; calyptra none.

I. Trichomes 5-8 mic. in diameter, forming a loose spiral 9-15 mic. in

diameter, the distance between the turns being 21-31 mic.

A. jenneri

SYNOPSIS OF THE BLUE-GREEN ALG7—MYXOPHYCEZ 217

II. Trichomes 2.5-3 mic. in diameter, forming a rather loose spiral about

6 mic, in diameter, the distance between the turns being 16-18

mic. A. gomontiana

Genus SPIRULINA

Plate XX, Fig. 35

Trichomes unicellular, cylindrical, without a sheath, forming a regu-

lar, more or less loose or close spiral; apex of trichome not tapering;

cell contents homogeneous or slightly granular.

I. Turns of the spiral not close together

1 Trichomes 1.2-1.8 mic. in diameter, forming a more or less loose,

somewhat irregular spiral 3.2-5 mic. in diameter, the distance

between the turns being 3-5 mic. S. meneghiniana

2 Trichomes 1.2-1.7 mic. in diameter, forming a somewhat loose,

regular spiral 2.5-4 mic. in diameter, the distance between the

turns being 2.7-5 mic. S. major

3. Trichomes 2 mic. in diameter, forming an especially regular spiral

5 mic. in diameter, the distance between the turns being 5 mic.

S. nordstedtii

4 Trichomes .6-.9 mic. in diameter, forming an especially regular

spiral 1.5-2.5 mic. in diameter, the distance between the turns

being 1.2-2 mic. S. subtilissima

5 Trichomes .4 mic. in diameter, forming an especially regular spiral

1.4-1.6 mic. in diameter, the distance between the turns being

1 mic. S. tenerrima

6 Trichomes .9 mic. in diameter, forming a very loose spiral 1.5 mic.

in diameter, the distance between the turns being 3.2 mic.

S. caldaria

Il. Turns of the spiral close together

1 Trichomes 1.2-1.8 mic. in diameter, forming a dense regular spiral

3-4.5 mic. in diameter, the turns being contiguous

S. versicolor

2 Trichomes 1-2 mic. in diameter, forming a somewhat irregular

dense or rarely regular spiral 3-5 mic. in diameter, the turns

being contiguous or nearly so S. subsalsa

III. Trichomes forming slender, flat, continuous bands (when untwisted

forming a complete ring), normally flattened and twisted, with

one to four or more turns S. duplex

Genus PHORMIDIUM

Plate XX, Fig. 36

Filaments simple, forming a woolly or felt-like layer or rarely float-

ing, attached at the base with free ends torn and ragged; sheaths thin,

transparent, mucous, agglutinated, partly or entirely diffluent; trichomes

218 JOSEPHINE E. TILDEN

cylindrical, in some species constricted at joints, never distinctly spiral;

apex of trichome often tapering, straight or curved, capitate or not capi-

tate; outer membrane of apical cell thickened into a calyptra in many

species.

I. Trichomes especially constricted at joints, even moniliform; apex of

trichome neither curved nor capitate

1 Trichomes scarcely 4 mic. in diameter

(1) Plants living in hot or in brackish water; trichomes 1.2-2.3

mic. in diameter; cells somewhat quadrate

| P. fragile

(2) Plants living in salt water; plant mass rose-colored; trichomes

1.7-2 mic. in diameter; cells longer than the diameter

P. persicinum

(3) Plants terrestrial, nestling in pits in rocks; trichomes 1.5 mic.

in diameter; cells somewhat quadrate P. foveolarum

2 Trichomes 6-8.5 mic. in diameter P. tinctorium

II. Trichomes rarely or scarcely constricted at joints; apex of trichome

straight or curved, capitate in many species

1 Trichomes scarcely 3 mic. in diameter

(1) Plant mass purplish violet, reddish brown or scarlet

A Filaments somewhat straight; trichomes slightly constricted

at joints; transverse. walls not granulated

P, luridum

B Filaments somewhat straight; trichomes fragile, frequently

interrupted, not constricted at joints; transverse walls

rarely visible P, rubrum

’ C Filaments very much twisted; trichomes not constricted at

joints; transverse walls marked by four protoplasmic

granules P, purpurascens

D Filaments curved, entangled or arranged parallel with each

other, trichomes not constricted at joints; apex of trichome

Straight, neither tapering nor capitate

P. crosbyanum

(2) Plant mass blue-green or olive

A Plants living in hot water; plant mass expanded, lamellose,

composed of many superposed papery layers; trichomes

.6-.8 mic. in diameter, not constricted at joints; apex of

trichome straight, not tapering P, treleasei

B Plant mass thin, membranaceous; trichomes 1-1.5 mic. in

diameter, not constricted at joints; apex of trichome

straight, tapering; transverse walls granulated

P, laminosum

SYNOPSIS OF THE BLUE-GREEN ALG7Z—MYXOPHYCEZ 219

C Plant mass thin, membranaceous; trichomes 1-2 mic. in diam-

eter, slightly constricted at joints; apex of trichome finally

becoming tapered and bent; transverse walls not granu-

lated P, tenue

D Plant mass thick, leathery; trichomes 2-2.5 mic. in diameter,

not constricted at joints; apex of trichome straight, obtuse

P. valderianum

E Plant mass lamellose; trichomes 2-2.8 mic. in diameter, slight-

ly constricted at joints; apex of trichome gradually taper-

ing, bent or twisted P. subuliforme

2 Trichomes more than 3 mic. in diameter

(1) Apex of trichome straight, not capitate

A Apical cell obtuse conical

a Plant mass encrusted with calcium carbonate

P, incrustatum

b Plant mass not encrusted with calcium carbonate

(a) Filaments somewhat straight; trichomes 3-5 mic. in

diameter; transverse walls covered by protoplasmic

granules P, inundatum

(b) Filaments flexuous; trichomes 3-4.5 mic. in diameter;

cells 3.4-8 mic. in length; transverse walls conspicu-

ous P. corium

eter; cells 2-4 mic. in length; transverse walls con-

spicuous P, papyraceum

(d) Plant mass membranaceous, mucous; trichomes 5-6.5

mic. in diameter, interrupted; apex of trichome

straight, obtuse; transverse walls sometimes finely

granulated P, interruptum

(e) Plant mass thin; trichomes 16-18 mic. in diameter, fre-

quently interrupted P, naveanum

B Apical cell not or scarcely tapering, truncate

a Sheaths thin, fragile, soon diffluent; trichomes 4.5-12 mic.

in diameter; cells 4-9 mic. in length P. retzii

b Sheaths firm, or mucous and diffluent, at times thick and

lamellose; trichomes 4-6 mic. in diameter; cells 1.5-2.7

mic. in length P, ambiguum

(2) Apex of trichome straight, capitate

A Trichomes slightly constricted at joints P. submembranace-

JOSEPHINE E. TILDEN

B Trichomes not constricted at joints

a Plants epiphytic, living in salt water; trichomes 4-4.5 mic.

in diameter, irregularly curved, very rarely straight; apex

of trichome gradually tapering; cells 4-11 mic. in length

P. laysanense

b Plants living in warm or fresh water; trichomes 4.5-9 mic.

in diameter, elongate, flexuous; apex of trichome gradu-

ally tapering; cells 3-7 mic. in length

P. favosum

ec Plants living in warm or fresh water; trichomes 6-8 mic. in

diameter, apex of trichome scarcely tapering; apical cell

oblique, depressed, conical P. calidum

d Plants living in fresh water; trichomes 5.5-11 mic. in diam-

eter, straight, fragile; apex of trichome briefly tapering;

apical cell straight, conical; cells 2-4 mic. in length

P. subfuscum

(3) Apex of trichome more or less curved, capitate

A Plant mass blue-green or dark brown

a Plants living in fresh water; trichomes 6-9 mic. in diameter;

apex of trichome curved or briefly spiraled

P. uncinatum

b Plants living on damp soil or on rocks, rarely under water;

trichomes 4-7 mic. in diameter; apex of trichome scarcely

curved, sometimes straight P. autumnale

B_ Plant mass dark purple P. setchellianum

Genus LYNGBYA

Plate XX, Fig. 37

Filaments free, unbranched, free-floating or forming a densely intri-

cate floccose or expanded mass; sheaths firm, of variable thickness, some-

times lamellose, colorless or rarely yellowish brown; trichomes some-

times constricted at the joints, either cbtuse or slightly tapering at the

apices; outer wall of apical cell sometimes thickened forming a calyptra.

Trichomes not more than 2 mic. in diameter

1 Plants living in salt water, epiphytic; transverse walls marked by

two refringent granules

(1) Filaments 1.5 mic. in diameter, trichomes .5 mic. in diameter,

cylindrical, not constricted at joints; cells 1.5 mic. in length

i L. mucicola

(2) Filaments 1.5-2 mic. in diameter; sheaths very thin, scarcely

visible; cells 2-7 mic. in length L. perelegans

SYNOPSIS OF THE BLUE-GREEN ALG—MYXOPHYCEZ 221

2 Plants living in fresh water

(1) Sheaths thin, colorless

A Filaments coiled or spiraled

a Filaments coiled; trichomes .8 mic. in diameter, constricted

at joints; cells 2.3-3.2 mic. in length

L. rivulariarum

b Filaments more or less regularly spiraled, sometimes

straight; trichomes 2 mic. in diameter; cells 1.2-3 mic. in

length L. lagerheimii

B Filaments straight or curved

a Filaments 1.9 mic. in diameter; apex of trichome bluntly

rounded; cells quadrate or a little longer than diameter

L. nana

b Filaments solitary and scattered; trichomes 1.5-1.8 mic. in

diameter, somewhat flexuous; cells up to 3.6 mic. in length

L. subtilis

c Filaments 1.8 mic. in diameter, at first attached, afterwards

free, short, straight or slightly curved; cells about equal

in length to the diameter L. distincta

(2) Sheaths more or less thick and gelatinous

A Plant mass ochre-yellow in color; sheaths at first thin, color-

less, later thick and yellowish; trichomes .9 mic. in diam-

eter, especially constricted at joints, frequently interrupted

L. ochracea

B Plant mass rust-colored; sheaths at first thin, colorless, later

thicker and rust-colored; trichomes .8-.9 mic. in diameter,

not constricted at joints, continuous

L. ferruginea

II. Trichomes 2-6 mic. in diameter; sheaths usually thin and colorless,

sometimes becoming thick and yellowish

1 Plants living in salt water, sometimes in brackish, fresh or hot

water

(1) Filaments coiled, densely entangled; sheaths thin, colorless,

later becoming thick and lamellose; trichomes 2.5-6 mic. in

diameter, not constricted at joints; cells 1.5-5.5 mic. in length

L. lutea

(2) Plants living in salt water, epiphytic; sheaths thin, delicate;

trichomes 3-4.5 mic. in diameter, constricted at joints; cells

up to 13 mic. in length L. holdenii

222 JOSEPHINE E. TILDEN

2 Plants. living in fresh water; sheaths usually thin and colorless

(1) Plant mass cespitose, light green; trichomes 2-3 mic. in diam-

eter, not constricted at joints; cells 1-3.7 mic. in length

L. digueti

(2) Plant mass cespitose, blue-green; sheaths delicate, smooth, |

usually inconspicuous; trichomes 3.2-3.5 mic. in diameter,

somewhat rigid, forming tufts L. penicillata

(3) Plant mass dull blue-green; trichomes 4-6 mic. in diameter, not

constricted at joints; cells 2.3-3 mic. in length

L. zrugineo-czru-

lea

(4) Plants epiphytic; filaments straight or sharply curved and

twisted; trichomes 5-6 mic. in diameter, not constricted at

joints L. cladophorz

(5). Plant mass at first adherent, afterwards free, rust-colored on

the outside, olive green within; sheaths colorless, sometimes

yellowish, slightly mucous and agglutinated, thick; trich-

omes 2.8-3.2 mic. in diameter, not constricted at joints; cells

2-6.4 mic. in length L. versicolor

III. Trichomes 5-60 mic. in diameter

1 Plants living in salt water, epiphytic; plant mass cespitose

(1) Plant mass purplish-violet; trichomes 5-8 mic. in diameter;

constricted at joints; cells 2.8-4.6 mic. in length; cell con-

tents rose-colored L. gracilis

(2) Plant mass dull blue-green; trichomes 6.5-8 mic. in diameter,

constricted at joints; cells 2-4 mic. in length; cell contents

pale blue-green L. meneghiniana

(3) Plant mass dark or dull yellowish green; trichomes 14-31 mic.

in diameter, evidently constricted at joints; cells 4-10 mic.

in length; cell contents frequently showing scattered coarse

granules, olive green L. sordida

2 Plants living in salt water; plant mass czspitose, extended, mu-

cous, dull yellowish or dark green, when dried becoming dark

violet; sheaths thick

(1) Trichomes 5-12 mic, in diameter; apex of trichome slightly

tapering, capitate; cells 2-3 mic. in length

L. semiplena

(2) Trichomes 9-25 mic. in diameter; apex of trichome not taper-

ing, not capitate; cells 2-4 mic. in length

L. confervoides

SYNOPSIS OF THE BLUE-GREEN ALGE—MYXOPHYCEX 223

3 Plants living in salt, brackish, fresh or warm water or on moist

earth

(1) Trichomes 8-24 mic. in diameter; apex of trichome slightly

tapering, capitate; apical cell truncate, rarely somewhat

acute-conical; cells 2.7-5.6 mic. in length

L. xstuarii

(2) Trichomes 16-60 mic. in diameter; apex of trichome not taper-

ing, not capitate; apical cell rotund; cells 2-4 mic. in length

L. majuscula

4 Plants living in fresh water, often in hot or warm water

(1) Plant mass cespitose, dull or dark green or blue-green

A Sheaths thickened and roughened with age; trichomes 6-10

_ mic. in diameter, not constricted at joints; apex of trichome

not tapering, not capitate; cells 1.7-3.3 mic. in length

L. martensiana

B_ Sheaths colorless, thin, papery; trichomes 7.5-13 mic. in diam-

eter, especially constricted at joints; apex of trichome not

tapering, not capitate; cells 3-10 mic. in length

L. putealis

C Sheaths colorless, thick, roughened; trichomes 11-16 mic. in

diameter, not constricted at joints; apex of trichome slight-

ly tapering, somewhat capitate; cells 2-3.4 mic. in length

L. major

(2) Plant mass floating, olive green; filaments forming a regular

loose spiral throughout the whole or a portion of their

length; trichomes 14-16 mic. in diameter, not constricted at

joints; cells 3.4-6.8 mic. in length L. spirulinoides

(3) Plant mass formed of loosely entangled filaments, dark green;

filaments 15-19 mic. in diameter; trichomes 12.5 mic. in

diameter; cells very short L. arachnoidea

Genus SYMPLOCA

Plate XX, Fig. 38

Filaments branched, ascending from a prostrate base, acai

together in erect or anastomosing fascicles, or wick-like bundles, more

or less procumbent, coalescing; false branches solitary; sheaths thin,

colorless, firm or somewhat mucous; apex of trichome straight, some-

times a little tapering; outer membrane of apical cell slightly thickened

in some species.

I. Plants living in salt water

1 Fascicles erect

(1) Plant mass blackish green; trichomes 4-6 mic. in diameter,

constricted at joints throughout entire length

S. atlantica

224 JOSEPHINE E. TILDEN

(2) Plant mass dull or dark lead-colored; trichomes 6-14 mic. in

diameter, constricted at joints near apices

S. hydnoides

constricted at joints S. lete-viridis

II. Plants living on moist earth, or in fresh or hot water

1 Trichomes 1-3 mic. in diameter |

(1) Plant mass blackish green; trichomes 4.6 mic. in diameter,

sometimes constricted at joints S. thermalis

(2) Plant mass compact, fibrous; trichomes 1.5-2.5 mic. in diam-

eter, not constricted at joints S. dubia

(3) Plant mass blue-green, changing to brown; trichomes 2-3 mic.

in diameter S. fuscescens

2 Trichomes 3-8 mic. in diameter

(1) Fascicles short, erect, spine-shaped; trichomes 3.4-4 mic. in

diameter, not constricted at joints; cells somewhat quadrate

or shorter than the diameter S. muralis

(2) Fascicles tapering from a broad base to a loose, somewhat

penicillate apex; trichomes 5.6 mic. in diameter; cells a little

longer than their diameter, after division shorter

S. borealis

(3) Fascicles elongate, usually procumbent, spine-shaped; trich-

omes 5-8 mic. in diameter,:not constricted at joints; cells

somewhat quadrate or longer than the diameter

S. muscorum

Genus PORPHYROSIPHON

Plate XX, Fig. 39

Filaments unbranched; sheaths firm, solid, lamellose, usually purple

or flesh-colored; trichomes solitary within the sheath; apical cell not

capitate.

Plant mass expanded, cushion-shaped, dark purple; sheaths firm,

finally becoming very thick, lamellose, with the apex tapering and

fibrillose; trichomes 8-19 mic. in diameter; cells 4.5-12 mic. in length

P, notarisii

Genus HYDROCOLEUS

Plate XX, Figs. 40, 41

Plant mass forming a cxespitose cushion, very rarely hardened with

calcium carbonate, or cxspitose but somewhat indefinite, or even not

at all cespitose, but Phormidium-like; sheaths always colorless, cylindri-

SYNOPSIS OF THE BLUE-GREEN ALG4—-MYXOPHYCE Zao

cal, somewhat lamellose, more or less mucous or somewhat amorphous,

later entirely diffluent; trichomes few within the sheath, often loosely

aggregated; apex of trichome straight, more or less tapering, capitate;

outer membrane of apical cell thickened into a calyptra; cells shorter than

the diameter of the trichome, in some species very short.

I. Plants living in salt water

1 Plant mass cxspitose

(1) Plant mass green becoming violet; sheaths cylindrical, moder-

ately mucous; trichomes 14-21 mic. in diameter

H. comoides

(2) Plant mass blackish green; sheaths irregular in outline,

strongly mucous; trichomes 18-24 mic. in diameter

H. cantharidosmus

2 Plant mass cespitose or forming an expanded mucous stratum,

blackish green; sheaths irregular in outline, strongly mucous or

even entirely diffluent; trichomes 8-16 mic. in diameter

H. lyngbyaceus

3 Plant mass mucous 7

(1) Plant mass yellowish brown or dull green; sheaths somewhat

amorphous or entirely diffluent;, trichomes 14-21 mic. in

diameter H. glutinosus

(2) Plant mass pale blue-green; sheaths agglutinated, forming a

diffuent, amorphous layer; trichomes 25-30 mic. in diameter

H. holdenii

II. Plants living in fresh water

1 Trichomes 6-8 mic. in diameter; apex of trichome gradually taper-

ing, evidently capitate; cells somewhat quadrate or one-half the

diameter of the trichome in length H. homeotrichus

2 Trichomes 12 mic. in diameter; cells quadrate or two or three

times shorter than the diameter H. ravenelii

3 Trichomes 16-19 mic. in diameter; apex of trichome somewhat

tapering, scarcely capitate; cells 2-5: times shorter than the

diameter H. heterotrichus

Genus HYPHEOTHRIX

Plate XXI, Fig. 42

Plants living on moist earth or dripping rocks; filaments prostrate,

commonly slightly branched, woven into a more or less compact mass;

sometimes hardened with calcium carbonate; sheaths always colorless.

I. Filaments very much twisted, scarcely flexible, ruptured if disen-

tangled

SYNOPSIS OF THE BLUE-GREEN ALGA!I—MYXOPHYCE Vt §

1 Plant mass thin, somewhat gelatinous, papery-membranaceous,

very hard when dry, not encrusted with calcium carbonate;

sheaths firm; trichomes 1-1.7 mic. in diameter, usually one or two

within the sheath H. calcicola

2 Plant mass flocculent, waving, light fawn-colored; filaments

1.2-1.8 mic. in diameter; sheaths inconspicuous; transverse walls

not visible H. hinnulea

3 Plant mass forming a small mat; filaments 1.5-2 mic. in diameter;

sheaths closely adherent, entirely diffluent

H. gleophila

4 Plant mass thin, cushion-shaped, mucous; filaments 1.8-2.2 mic. in

diameter; sheaths close H. herbacea

5 Plant mass sometimes expanded, forming loosely interwoven

masses or small cushion-shaped clusters; filaments 3.5-4 mic. in

diameter; sheaths firm, close H. tenax

6 Plant mass somewhat spherical, hollow, tough, yellowish or light

straw-colored; filaments 4-6 mic. in diameter; trichomes 1.5-2

mic. in diameter H. bullosa

7 Plant mass compact, leathery, brick-colored; filaments up to 7.5

mic. in diameter; sheaths wide, membranaceous, firm, homogene-

ous, smooth; trichomes 3.2-4 mic. in diameter, here and there in-

terrupted H. turicensis

8 Plant mass more or less expanded, olive green; filaments 8-11

mic. in diameter; sheaths moderately wide; trichomes 3.5 mic.

in diameter, here and there interrupted, often constricted at

joints H. aikensis

II. Filaments long and flexible, disentangled without rupturing

1 Plant mass encrusted with calcium carbonate; trichomes 1-1.7 mic.

in diameter; cells longer than the diameter

H. coriacea

2 Plant mass not encrusted with calcium carbonate; trichomes 1.5-2

mic. in diameter; cells longer than the diameter

H. lardacea

3 Plant mass not encrusted with calcium carbonate; trichomes 1.5-3

mic. in diameter; cells longer than the diameter

H. arenaria

4 Plant mass compact, leathery, roughened; trichomes 2.3-2.8 mic.

in diameter; cells a little shorter than the diameter

H. vulpina

228 JOSEPHINE E. TILDEN

5 Plant mass membranaceous, firm, smooth, pale rose or dark red;

trichomes 5.6-8.3 mic. in diameter; cells somewhat quadrate

H. pallida

Genus SYMPLOCASTRUM

Plate XXI, Fig. 43, 44, 45

Plants terrestrial or living on damp rocks; filaments twisted and en-

tangled, ascending from a prostrate base, agglutinated together in erect

fascicles; sheaths colorless.

I. Plant mass blue-green; trichomes 1.4-2 mic. in diameter, constricted

at the joints; cells shorter than the diameter

S. fragile

II. Plant mass flesh-colored or reddish; trichomes 1.6-2 mic. in diam- -

eter; cells usually longer than the diameter

S. rubrum

III. Plant mass gray or yellowish; trichomes 1.9-2.3 mic. in diameter;

cells longer than the diameter S. cuspidatum

IV. Plant mass blackish, olive or lead-colored; trichomes 3-6 mic. in

diameter; cells somewhat quadrate or longer than the diameter

S. friesii

Genus INACTIS

Plate XXI, Fig. 46

Plants growing in moist places or in rivers; filaments cespitose,

often with numerous false branches, forming cushions which finally often

become encrusted with calcium carbonate and hardened, zonate within,

or aggregated into penicillate, floating fascicles; sheaths colorless or

nearly so. ;

I. Plant mass cushion-shaped, tufted

1 Plant mass strongly encrusted with calcium carbonate, stony; fila-

ments straight, somewhat simple; trichomes 1-2 mic. in diam-

eter; cells somewhat quadrate I. pulvinata

2 Plant mass strongly encrusted with calcium carbonate, stony; fila-

ments slender, simple in basal portions, fasciculately branched

above; trichomes 1.4-3 mic. in diameter; cells somewhat quadrate

or longer than the diameter I. fasciculata

3 Plant mass cushion-shaped or crustaceous, not hardened with cal-

cium carbonate; filaments forming trunk at base, very much

branched in upper portions; trichomes 1-1.5 mic. in diameter;

cells longer than the diameter I. lacustris

4 Plant mass somewhat hemispherical, plano-convex; filaments more

or less branched, growing in tufts; cells two or three times

longer than broad I. austini

SYNOPSIS OF THE BLUE-GREEN ALG7Z—MYXOPHYCEZ 229

II. Plant mass forming penicillate fascicles, floating

1 Plant mass submerged, attached; filaments very long; trichomes

1.4-2.4 mic. in diameter, constricted at joints

I. tinctoria

2 Plant mass submerged, epiphytic on other alge; trichomes 3-6

mic. in diameter, usually constricted at joints

I. simmonsize

3 Plant mass submerged, attached; filaments very long; trichomes

6 mic. in diameter, constricted at joints I, mexicana

III. Filaments solitary, growing in gelatinous mass formed by other

alge; trichomes 1.5-2 mic. in diameter, not constricted at joints

I, hawaiensis

Genus SCHIZOTHRIX

Plate XXI, Fig. 47

Plants living on moist earth or in water, or in inundated places,

rarely entirely aquatic; filaments forming erect or prostrate, Symploca-

like fascicles or a pannose stratum, rarely floating free; sheaths in the

beginning colorless, finally becoming yellowish brown, purplish pink or

bluish.

I. Cells somewhat quadrate or shorter than the diameter

1 Plant mass thin, encrusted, often widely expanded or in tangled

tufts among other alge; sheaths colorless, very transparent;

trichomes 1-1.5 mic. in diameter, constricted at joints; cells

somewhat quadrate S. hyalina

2 Plant mass cespitose or appressed, semiorbicular; sheaths very

thick, lamellose; trichomes 4-9 mic. in diameter, usually solitary

within the sheath S. thelephoroides

3 Plant mass indefinite, sheaths purple, orange or rose-colored;

trichomes 6-8 mic. in diameter, many within the sheath

S. purpurascens

4 Plant mass indefinite, woolly, lead-colored; sheaths very thick,

lamellose; trichomes, 7.5-8.5 mic. in diameter

S. chalybea

5 Plant mass not cespitose; sheaths yellowish orange; trichomes

7-13 mic. in diameter; cells somewhat quadrate or twice as short

as the diameter S. muelleri

II. Cells longer than the diameter

1 Filaments very long; sheaths dark lead-colored, irregular in out-

line; trichomes 1.7 mic. in diameter S. braunii

230 JOSEPHINE E. TILDEN

2 Filaménts forming a loose, cobwebby mass within sandstone rock;

sheaths cylindrical, rough, usually colorless and not lamellose,

sometimes brownish and lamellose; trichomes 3.5-4.8 mic. in

diameter; cells quadrate or a little longer than the diameter

S. rupicola

Genus DASYGLC@A

Plate XXI, Fig. 48

Sheaths very wide, colorless or yellowish brown; trichomes very few

within the sheath, very loosely aggregated; apex of trichome straight, not

capitate; cells often longer than the diameter.

Plant mass amorphous, gelatinous; filaments twisted, entangled,

divided into fringes at the apex; sheaths sometimes somewhat lamellose;

trichomes 4-6 mic. in diameter, constricted at joints; apex of trichome

sometimes very gradually tapered; cells 4-13 mic. in length; cell contents

coarsely granular D. amorpha

Genus MICROCOLEUS

Plate XXI, Fig. 49

Plants living on soit’ in fresh water or sometimes in salt water; fila-

ments simple or vaguely branched, creeping on the ground, sometimes

growing among other algx; sheaths colorless, more or less regularly

cylindrical, not lamellose, in many species finally diffluent; trichomes

many within the sheath in well developed filaments, closely crowded,

often twisted into rope-like bundles; apex of trichome straight, tapering;

apical cell acute, rarely obtuse conical, in one species capitate.

I. Plants living in salt water; apical cell not capitate, pointed

1 Trichomes 1.5-2 mic. in diameter, constricted at joints

M. tenerrimus

2 Trichomes 2.5-6 mic. in diameter, constricted at joints

M. chthonoplastes

II. Plants living on soil; apical cell capitate M. vaginatus

III. Plants living in fresh water; apical cell not capitate

1 Sheaths mucous, diffuent; trichomes 4-5 mic. in diameter, especial-

ly constricted at joints M. lacustris

2 Sheaths somewhat mucous, not or scarcely diffluent; trichomes

5-7 mic. in diameter, not constricted at joints

M. paludosus

3 Plant mass large, cushion-like; trichomes 5-6 mic. in diameter

M. pulvinatus

4 Sheaths very mucous and agglutinated; trichomes 6-10 mic. in

diameter, especially constricted at joints M. subtorulosus

SYNOPSIS OF THE BLUE-GREEN ALGA7—MYXOPHYCE# 231

Genus CATAGNYMENE

Plate XXI, Fig. 50

Filaments multicellular, floating free, surrounded by thin, close

sheaths, enclosed in widely expanded, gelatinous diffuent envelopes, sep-

arating easily into fragments through the death of cells.

I. Gelatinous envelope 93-100 mic. in diameter; trichomes up to 16 mic.

in diameter, straight or curved C. pelagica

11. Gelatinous envelope 150-168 mic. in diameter; trichomes 20-22 mic.

in diameter, spirally coiled C. spiralis

Genus HALIARACHNE

Filaments multicellular, floating free, in somewhat globose or elon-

gate, gelatinous colonies, arranged in two layers, radiating from the cen-

ter, hooked at the apex; reproduction by division of the colony.

Colony lenticular, 450-700 mic. in diameter; apical cell possessing a

calyptra; cells about 8 mic. in diameter, 4-7 mic. in length; cell contents

showing gas vacuoles H. lenticularis

Family II. NOSTOCACEZE

Sheaths forming a more or less distinct mucous, gelatinous or mem-

branaceous tegument, mostly confluent, often not present; trichomes con-

sisting of a single row of uniform cells, with heterocysts, usually twisting

and entangled, not branched, showing no differentiation of base and apex;

reproduction by means of vegetative division, hormogones and gonidia.

I. Sheaths inconspicuous, or mucous and diffluent, or gelatinous, firm

and thick

1 Trichomes flexuous and contorted, forming a plant mass or colony

of definite shape

(1) Colony usually of a rounded or expanded character, gelati-

nous, made up of dissolved individual sheaths, attached to

the substratum or floating free in water; heterocysts inter-

calary Nostoc

Plate XXI, Fig. 51

(2) Colony tubular, cylindrical; filaments somewhat straight,

parallel, agglutinated Wollea

Plate XXI, Fig. 52

2 Trichomes more or less straight, free, or forming a thin mucous

layer of indefinite shape

(1) Heterocysts and gonidia intercalary

A Trichomes free; cells disc-shaped; shorter than their diam-

eter; gonidia seriate, remote from the heterocysts

Nodularia

Fig. 53

232 JOSEPHINE E. TILDEN

B. Trichomes naked, or with a thin mucous sheath, free or ag-

gregated without order to form a flocculent mass; cells

equal to or longer than their diameter; gonidia solitary, in

pairs or in short series Anabzna

Plate XXII, Fig. 54

C Trichomes short, aggregated in parallel bundles to form thin,

feathery, plate-like masses Aphanizomenon

Fig. 55

(2) Heterocysts terminal and the gonidia always contiguous to

them Cylindrospermum

Fig. 56

II. Sheaths thin, membranaceous, persistent; filaments free or agglutin-

ated in a parallel manner

1 Sheaths not present; trichomes single, endophytic; heterocysts

terminal Richelia

Fig. 57

2 Trichomes single within the sheath; heterocysts intercalary

Aulosira

Fig. 58

3 Trichomes single within the sheath; heterocysts intercalary and

terminal Microchete

Plate XXIII, Fig. 59

4 Trichomes usuaily many within the sheath, forming a membran-

aceous or filamentous mass Hormothamnion

Fig. 60

Genus NOSTOC

Plate XXI, Fig. 51

Plant mass or colony at first globose or oblong, afterwards assum-

ing various forms (globose, foliose, filiform, bullose) in different species,

solid or hollow, mucous, gelatinous or leathery, made up of tangled trich-

omes and their more or less dissolved sheaths; filaments flexuous, curved,

entangled, coalesced; sheaths sometimes distinct, sometimes invisible;

trichomes often torulose; cells depressed spherical, barrel-shaped or cylin-

drical; heterocysts intercalary and (in younger plants) terminal; gonidia

spherical or oblong, developed centrifugally in series between the hetero-

cysts.

I. Plants living in fresh water; forming minute, disc-shaped specks or

patches on aquatic plants; plant mass growing at the periphery;

filaments closely contorted N. cuticulare

II. Plants living in fresh water, microscopic, granular, aggregated, hav—

ing the appearance of Aphanocapsa; filaments very closely en-

tangled; trichomes scarcely distinct N. punctiforme

SYNOPSIS OF THE BLUE-GREEN ALG#—MYXOPHYCEZ 233

III. Plants living in fresh water, very minute; trichomes 2-3.5 mic. in

diameter, distinct

1 Plant mass very minute, punctiform; filaments loosely flexuous;

trichomes 3-3.5 mic. in diameter; gonidia about 4 mic. in diam-

eter, 6-8 mic. in length, oblong N. paludosum

2 Plant mass small, adherent, somewhat globose; orange or green;

trichomes 2-2.5 mic. in diameter, very short, strongly curved

N. aureum

3 Plant mass small, gelatinous, membranaceous, soft, green, blue-

green or brownish; trichomes 3-4 mic. in diameter, flexuously

curved, somewhat densely entangled N. comminutum

IV. Plants living in fresh water; plant mass large, gelatinous, fragile,

at first spherical, afterwards becoming torn and irregularly ex-

panded

1 Filaments numerous, abruptly contorted, entangled; trichomes

3.5-4 mic. in diameter; gonidia 6-7 mic. in diameter, 7-8 mic. in

length N. linckia

2 Filaments flexuous, loosely entangled

(1) Gonidia 6-7 mic. in diameter, spherical; wall of gonidium

smooth; trichomes 4 mic. in diameter N. piscinale

(2) Gonidia olong; wall of gonidium smooth

A Trichomes 4-4.2 mic. in diameter; gonidia 6-8 mic. in diam-

eter, 7-10 mic. in length, contiguous; wall of gonidium be-

coming brownish or colorless N. rivulare

B Trichomes 3.5-4 mic. in diameter; gonidia 6 mic. in diameter,

8-10 mic. in length, not contiguous; wall of gonidium color-

less N. carneum

C Trichomes 4 mic. in diameter; cells different in shape, some

cylindrical, others barrel-shaped or spherical depressed;

gonidia 6-7 mic. in diameter, 10-12 mic. in length, not con-

tiguous; wall of gonidium colorless or becoming yellowish

N. spongizforme

V. Plants living on soil; colonies gelatinous, soft, at first spherical,

soon confluent and flattened, attached to soil or mosses

1 Cells cylindrical; trichomes 4 mic. in diameter; gonidia 6-8 mic. in

diameter

(1) Gonidia 14-19 mic. in length; wall of gonidium smooth

N. ellipsosporum

(2) Gonidia 8-14 mic. in length; wall of gonidium furnished with

short spines N. gelatinosum

234 JOSEPHINE E. TILDEN

2 Cells oval, spherical or spherical depressed

(1) Trichomes 3-4 mic. in diameter; gonidia 4-8 mic. in diameter,

8-12 mic. in length, oblong, in a catenate series

N. muscorum

(2) Trichomes 2.2-3 mic. in diameter; gonidia 4 mic. in diameter,

6 mic. in length, oval N. humifusum

VI. Plants living on soil, sometimes submerged; colonies free, at first

spherical, then expanding symmetrically or irregularly; cells

somewhat globose

1 Colonies gelatinous, spongy, lacunose, somewhat pellucid, green,

olive or brownish; trichomes 4 mic. in diameter; gonidia 7 mic.

in diameter, 7-10 mic. in length, often oval; wall of gonidium

smooth, colorless N. foliaceum

2 Colonies expanded, irregular or orbicular, very thin, small, mem-

branaceous, pellucid, blue-green; trichomes 4 mic. in diameter

N. punctatum

3 Colonies at first spherical, afterwards becoming flattened and final-

ly spreading out into irregular, membranaceous sheets; sur-

rounded by a firm outer layer; trichomes 4-5.6 mic. in diameter

N. commune

4 Colonies free, spherical, becoming irregularly plicate-tuberculate,

thick, solid, surrounded by a firm outer layer; trichomes 4-5 mic.

in diameter; gonidia 5 mic. in diameter, 7 mic. in length, oval;

wall of gonidium thick, smooth, becoming brownish

N. sphericum

5 Colonies spherical, finally becoming flattened, membranaceous;

trichomes 2.5-3 mic. in diameter N. minutum

6 Plants living in hot water; colonies indefinitely expanded, lacini-

ate; filaments 2 mic. (?) in diameter N. calidarium

7 Colonies somewhat spherical, small, very hard, sometimes soft,

with surface often corrugated; trichomes 6.5-8.2 mic. in diam-

eter N. austinii

VII. Plants living on soil or in fresh water; colonies spherical, sur-

rounded by a firm outer layer

1 Plants living on soil

(1) Colonies small; trichomes 8-9 mic. in diameter; gonidia some-

what spherical, two or three times larger than the cells; wall

of gonidium thin, very smooth N. macrosporum

(2) Colonies spherical or oblong, rarely beyond 1 cm. in diam-

eter, somewhat pellucid; trichomes 5-8 mic. in diameter;

gonidia 6-7 mic. in diameter, 9-15 mic. in length, oval

N. microscopicum

SYNOPSIS OF THE BLUE-GREEN ALG——MYXOPHYCEH% 235

(3) Colonies small or of medium size, spherical; trichomes 4-7

mic. in diameter; gonidia 6-7 mic. in diameter, exactly spher-

ical; wall of gonidium somewhat thick, rough

N. spheroides

2 Plants living in fresh water

(1) Colonies irregularly somewhat orbicular, gregarious and some-

times aggregated; trichomes 5 mic. in diameter

N. depressum

(2) Colonies spherical, usually aggregated in grape-like clusters;

trichomes 3.5-4 mic. in diameter N. glomeratum

(3) Colonies gregarious, pellucid, sky blue or blue-green; trich-

omes 5-7 mic. in diameter; cells barrel-shaped

N. czruleum

(4) Colonies spherical, surrounded by a leathery outer layer;

trichomes 4-6 mic. in diameter N. pruniforme

VIII. Plants living in fresh water, attached; colonies somewhat spher-

ical, bullate, rarely disc-shaped, surrounded by a firm outer

layer; trichomes slender

1 Trichomes 3-3.5 mic. in diameter, especially cylindrical; gonidia 5

mic. in diameter, 7 mic. in length; wall of gonidium smooth

N. verrucosum

2 Trichomes 2-3 mic. in diameter, distinctly torulose; gonidia 3-4

mic. in diameter, 5-6 mic. in length; wall of gonidium smooth,

brown N. amplissimum

3 Filaments radiating from the center, flexuous, very densely

twisted and entangled near the surface; trichomes 4-4.5 mic. in

diameter; gonidia 4-5 mic. in diameter, 7-8 mic. in length, oval;

wall of gonidium smooth, yellowish N. parmelioides

Genus WOLLEA

Plate XXI, Fig. 52

Plant mass or colony tubular, cylindrical, somewhat membranace-

ous, soft; filaments somewhat straight, parallel or slightly curved, ag-

glutinated; sheaths confluent; heterocysts intercalary; gonidia catenate,

contiguous to the heterocysts or remote from them.

Trichomes 4-5 mic. in diameter, numerous, erect, parallel or slightly

curved; cells oblong or cylindrical, closely connected; heterocysts 6 mic.

in diameter, oval or somewhat spherical, yellow or pale orange; gonidia 7

mic. in diameter, 15-22 mic. in length, numerous, cylindrical, catenate

W. saccata

236 JOSEPHINE E. TILDEN

Genus NODULARIA

Plate XXI, Fig. 53

Filaments free; sheaths colorless, close, usually thin, mucous, some-

times diffluent; trichomes more or less straight; cells short, depressed, .

disc-shaped; heterocysts depressed; gonidia spherical, somewhat spher-

ical or disc-shaped, developed in series between the heterocysts; wall of

gonidium smooth.

I. Trichomes less than 8 mic. in diameter

1 Filaments 4-6 mic. in diameter; gonidia 6-8 mic. in diameter, some-

what spherical N. harveyana

2 Filaments 6-7 mic. in diameter; gonidia 7-10 mic. in diameter,

spherical depressed N. spherocarpa

3 Trichomes 6-8 mic. in diameter; cells short, about half as long as

wide N. paludosa

II. Filaments more than 8 mic. in diameter

1 Trichomes 7.5-9.5 mic. in diameter; cells nearly as long as broad

before division N. hawaiiensis

2 Filaments 10-11 mic. in diameter; gonidia 10-12 mic. in diameter, 9

mic. in length, spherical depressed, in series

N. armorica

3 Filaments 8-18 mic. in diameter; gonidia 12-15 mic. in diameter,

6-10 mic. in length; somewhat spherical or elliptical

N. spumigena

4 Trichomes (?) 33-38 mic. in diameter; cells short

N. mainensis

Genus ANABZAINA

Plate XXII, Fig. 54

Sheaths not present or when present often diffluent; trichomes equal

throughout or tapering at the apices, usually rigid and fragile, sometimes

circinate, free or aggregated without order to form a flocculent mass; cells

equal to or longer than their diameter; apical cells sometimes conical;

heterocysts numerous and intercalary; gonidia variously disposed, some-

times solitary, sometimes lying on each side of a heterocyst, rarely in

short catenate series.

I. Gonidia oval or spherical

1 Gonidia oval or barrel-shaped, remote from the heterocysts in

catenate series

(1) Wall of gonidium smooth A. variabilis

(2) Wall of gonidium papillose A. hallensis

2 Gonidia spherical, contiguous to heterocysts, solitary or in short

series, 12-20 mic. in diameter A. spherica

SYNOPSIS OF THE BLUE-GREEN ALGAE—MYXOPHYCE 237

II. Gonidia variously disposed, sometimes contiguous to heterocysts,

sometimes remote from them, cylindrical, straight or curved

1 Trichomes usually circinate; gonidia curved, obliquely truncate at

the apices

(1) Gonidia 7-13 mic. in diameter, 20-50 mic. in length, curved,

oblique, inequilateral, contiguous to or rarely remote from

the heterocysts; wall of gonidium smooth, colorless or yel-

lowish; trichomes 4-8 mic. in diameter

A. flos-aquz

(2) Gonidia 16-18 mic. in diameter, up to 30 mic. in length, curved,

oblique or cylindrical, the younger ones somewhat spherical,

usually remote from the heterocysts; wall of gonidium

smooth, colorless; trichomes 8-14 mic. in diameter

A. circinalis

2 Trichomes straight; gonidia cylindrical, straight, usually remote

from the heterocysts, solitary or in series

(1) Trichomes 4-5 mic. in diameter; sheaths sometimes present;

gonidia 14-17 mic. in length A, inzqualis

(2) Trichomes 5-8 mic. in diameter; sheaths occasionally present;

gonidia 7-10 mic. in diameter, up to 30 mic. and more in

length A. catenula

(3) Trichomes 4.2-6 mic. in diameter; sheaths present; gonidia 6

mic. in diameter, 14-20 mic. in length A. laxa

III. Gonidia contiguous to heterocysts on each side, developed centripe-

tally, cylindrical or somewhat cylindrical

1 Gonidia 7-12 mic. in diameter, 18-28 mic. in length, short, some-

what cylindrical, often slightly constricted in the center; apical

cell conical A. torulosa

2 Gonidia 8-10 mic. in diameter, 20-40 mic. in length, especially cylin-

drical; apical cells obtuse A. oscillarioides

3 Gonidia 15-20 mic. in diameter, 50-90 mic. in length, cylindrical or

more commonly tapering slightly from the middle to the

rounded ends A. bornetiana

Genus APHANIZOMENON

Plate XXII, Fig. 55

Colonies thin, feathery, plate-like or spindle-shaped bundles, blue-

green, floating; sheaths not present; trichomes short, tapering at the ends,

agglutinated; heterocysts scattered; gonidia cylindrical, much elongated,

solitary, developed sparingly between the heterocysts.

Colonies small, aggregated in membranaceous flakes, fragile; trich-

omes 5-6 mic. in diameter, rigid; cells somewhat quadrate, 5-15 mic. in

238 JOSEPHINE E. TILDEN

length; heterocysts 6-7 mic. in diameter, 15-20 mic. in length, somewhat

cylindrical; gonidia 7-8 mic. in diameter, 60-80 mic. in length, containing

granular protoplasm; wall of gonidium smooth, colorless

A. flos-aquz

Genus CYLINDROSPERMUM

Plate XXII, Fig. 56 ,

Plant mass expanded, indefinite, mucous; sheaths not present; trich-

omes equal, short, embedded in an amorphous mucus; cells cylindrical,

longer than their diameter; heterocysts terminal, solitary; gonidia devel-

oped from the cell or cells next the heterocyst, generally solitary, rarely

seriate.

I. Gonidia solitary

1 Gonidia cylindrical, up to 40 mic. in length

(1) Gonidia 10-16 mic. in diameter, 32-40 mic. in length

C. stagnale

(2) Gonidia 11-12 mic. in diameter, 23-24 mic. in length

C, comatum

2 Gonidia oblong or ventricose-elliptical

(1) Wall of gonidium punctate

A Gonidia 10-15 mic. in diameter, 20-38 mic. in length, ventri-

cose-elliptical; wall of mature gonidium rough, punctate

C. majus

B_ Gonidia 6-6.5 mic. in diameter, 16-19 mic. in length, elliptical;

wall of gonidium very finely granular

C. minutum

(2) Wall of gonidium smooth

A Gonidia 8-9 mic. in diameter, 18-20 mic. in length

C. minutissimum

B_ Gonidia 9-12 mic. in diameter, 18-20 mic. in length

C. muscicola

C Gonidia 12-14 mic. in diameter, 20-38 mic. in length

C. licheniforme

II. Gonidia seriate C. catenatum

Plate XXII, Fig. 57

Sheaths not present; trichomes single, endophytic; heterocysts soli-

tary, situated at the base of the trichome.

Trichomes 5.6-9-8 mic. in diameter, 50-105 mic. in length, short,

straight or nearly straight, thickened at the apices; heterocysts 9.8-11.2

mic. in diameter R. intracellularis

SYNOPSIS OF THE BLUE-GREEN ALG#—MYXOPHYCE® 239

Genus AULOSIRA

Plate XXII, Fig. 58

Filaments free, equal, scattered or in fascicles; sheaths membranace-

ous, close; cells cylindrical or barrel-shaped; heterocysts intercalary; gon- —

idia developed at intervals between the heterocysts, remote from or conti-

guous to them, cylindrical, in catenate series.

Filaments 10-11 mic. in diameter; trichomes 9.5 mic. in diameter;

cells 3 mic. in length; heterocysts usually 9.5 mic, in diameter, 11 mic.

in length, always intercalary A. schauinslandii

Genus MICROCHZETE

Plate XXIII, Fig. 59

Plants small, living in fresh or salt water, aggregated into star-

shaped or cushion-shaped tufts; filaments unbranched, erect, attached at

the base; sheaths present; trichomes single within the sheath; heterocysts

basal and intercalary; gonidia developed from the lower cells.

I. Plants living in fresh water; heterocysts basal and intercalary

1 Filaments 4.4-5.1 mic. in diameter; sheaths colorless, wide

M. tenuissima

2 Filaments 10 mic. in diameter; sheaths simple, thin, close

M. tenera

3 Filaments 16-18 mic. in diameter; sheaths at first thin, later be-

coming lamellose, colorless M. robusta

II. Plants living in salt water; heterocysts basal

1 Plant mass densely cespitose; filaments 6-7 mic. in diameter, thick-

ened into a bulb at the base M. grisea

2 Plant mass loosely cespitose; filaments 7-9 mic. in diameter, flex-

uous, scarcely thickened at base M. vitiensis

Genus HORMOTHAMNION

Plate XXIII, Fig. 60

Plant mass formed from filaments growing together in a longitudinal

manner, sometimes developing as an expanded layer, sometimes erect,

filiform, torn and branched, not surrounded by a common gelatinous tegu-

ment; sheaths membranaceous, thin, often diffluent, colorless; trichomes

moniliform; usually many within the sheath; heterocysts intercalary; go-

nidia not known.

I. Plant mass floccose, entangled; trichomes 9-12 mic. in diameter

H. solutum

II. Plant mass erect, cxspitose, resembling Symploca; trichomes 6-7

mic. in diameter H. enteromorphoi-

des

Wein are

uisigiet

a RS

Pie pS

PLATE XXII

SYNOPSIS OF THE BLUE-GREEN ALGH—MYXOPHYCEZ 241

Family III. SCYTONEMACEZ:

Filaments branched; false branches formed by the perforation of the

sheath by the trichome which thereupon issues as one or two long, flex-

uous branches each developing a sheath of its own; sheaths homogeneous

and colorless, or lamellose and yellowish or brownish, firm, tubular; trich-

omes consisting of a single row of cells, one or more included in a sheath;

heterocysts and gonidia variously disposed; reproduction by means of

vegetative division, hormogones and gonidia.

I. Trichomes single within the sheath

1 Heterocysts not present; filaments free or forming felt-like masses,

branched; false branches often in pairs Plectonema

Plate XXIV, Fig. 61

2 Heterocysts present

(1) False branches usually arising between two heterocysts, single

or in pairs; sheaths delicate or very thick, parallel, or more

or less diverging towards the apex Scytonema

Plate XXV, Fig. 62

(2) False branches usually arising in the immediate region of the

heterocysts, single; sheaths somewhat thin, flexible, more or

less fragile Tolypothrix

Fig. 63

II. Trichomes or filaments several within the sheath

1 Filaments straight, associated in tufts; sheaths thin; trichomes

two or more within the sheath; heterocysts basal

Desmonema

Fig. 64

2 Filaments several contorted within a common tegument, asso-

ciated in a gelatinous stratum; trichomes single within the

sheath Diplocolon

Fig. 65

Genus PLECTONEMA

Plate XXIII, Fig. 61

Filaments free or forming felt-like masses, branched; false branches

solitary or in pairs; sheaths firm, colorless or rarely yellowish orange;

trichomes frequently constricted at the joints; apex of trichome straight,

very rarely tapering; calyptra none.

I. Plants large, cespitose; trichomes 3 mic. and more in diameter

1 Plant mass cespitose, rotund, light green; trichomes 5-10 mic. in

diameter, here and there constricted at joints

P, tenue

2 Plant mass cespitose, indefinite, brownish green; trichomes 11-22

mic. in diameter P, tomasinianum

Q42Py a

NOSE NE BN eres

SCE IR gs

PiLate XXIII

SYNOPSIS OF THE BLUE-GREEN ALGA:—MYXOPHYCE® 243

3 Plant mass widely expanded, indefinite, blackish, rarely yellowish

green; trichomes 28-47 mic. in diameter, not constricted at

joints P. wollei

II. Plant mass very thin, not cespitose; trichomes 1-4 mic. in diam-

eter

1 Filaments somewhat flexuous, immersed in dead shells; trichomes

9-1.5 mic. in diameter, not constricted at joints

P. terebrans

2 Filaments somewhat straight, growing among various gelatinous

algw; trichomes 1-1.5 mic. in diameter P. nostocorum

3 Filaments usually strongly flexuous, densely entangled in a rose-

colored membrane; trichomes 1.2-1.8 mic. in diameter

P, roseolum

4 Filaments long, entangled, flexuous, much branched, forming a

rose-colored or reddish brown mass adhering to rocks or larger

alge; trichomes 1.2-2 mic. in diameter P. golenkinianum

5 Filaments very long, entangled in dense balls; trichomes 2-2.5 mic.

in diameter P. calothrichoides

6 Filaments long, flexuous, much branched, forming a black or

brownish green mass; trichomes 2-3.5 mic. in diameter

P. battersii

Genus SCYTONEMA

Plate XXIV, Fig. 62

Filaments branched; false branches usually arising between two

heterocysts, solitary or in pairs, formed by the lateral perforation of the

sheath by the trichome; trichomes single within the sheath, straight;

hormogones terminal, solitary; gonidia spherical or oval, observed in a

few species; wall of gonidium thin, smooth.

I. Sheaths homogeneous or formed of parallel layers

1 Plants living in fresh water

(1) Filaments 5-8 mic. in diameter S. conchophilum

(2) Filaments 12-16 mic. in diameter S. arcangelii

(3) Filaments 18-24 mic. in diameter S. coactile

(4) Filaments about 30 mic. in diameter S. rivulare

(5) Filaments 36 mic. in diameter S. occidentale

(6) Filaments 16-36 mic. in diameter S. crispum >

2 Plants living in warm water

(1) Filaments 16 mic. in diameter S. caldarium

(2) Filaments 25 mic. in diameter S. azureum

eet

EAT

AUT

ebs¢ z

ae La Ls

STEELE

PLaTE XXIV

SYNOPSIS OF THE BLUE-GREEN ALGE—MYXOPHYCE

245

3 Plants living on soil, rocks, or bark, not submerged

(1)

(2)

(3)

(4)

(5)

(6)

(7)

Filaments

7-15 mic. in diameter

9-15 mic. in diameter

12-15 mic. in diameter

10-18 mic. in diameter

up to 20 mic. in diameter

15-20 mic. in diameter

S. hofmanni

S. varium

S. javanicum

S. ocellatum

S. intertextum

S. austinii

15-21 mic. in diameter; cells compressed

wm Pk WH eK

ott.

S. millei

(8) Filaments 15-21 mic. in diameter; cells somewhat quadrate or

elongate

(9) Filaments 19-24 mic. in diameter

(10) Filaments 20-25 mic. in diameter

(11) Filaments 16-30 mic. in diameter

soil or rocks, not submerged

Filaments

10-15 mic.

12-18 mic.

15-21 mic.

18-36 mic.

40-75 mic.

Sheaths thick, lamellose, forming wings or membranaceous expan-

in diameter

S. guyanense

S. amplum

S. wolleanum

S. stuposum

Sheaths lamellose, with diverging layers; plants usually living on

S. tolypotrichoides

S. flavo-viride

S. mirabile

S. myochrous

S. badium

sions (ocree); branches in basal portion of filament issuing in

pairs, those in the upper portion solitary

1 Plants living in water

(1) Plants living in salt water; filaments 28-50 mic. in diameter

2 Plants living on damp rocks or on bark

S. fuliginosum

(2) Plants living in fresh water, on dripping rocks or submerged

(1) Filaments 12-16 mic. in diameter

(2) Filaments 15-30 mic. in diameter

(3) Filaments 24-40 mic. in diameter

Genus TOLYPOTHRIX

Plate XXIV, Fig. 63

S. alatum

S. junipericolum

S. crustaceum

S. densum

Filaments branched; false branches usually arising in the immediate

region of the heterocysts, rarely between two heterocysts, single; sheaths

246 JOSEPHINE E. TILDEN

somewhat thin, flexible, more or less fragile; gonidia spherical, oval or

elliptical, often many in a series; wall of gonidium smooth, thin.

I. Sheaths thin

1 Plants living in water

(1) Filaments 8-10 mic. in diameter T. tenuis

(2) Filaments 9-12.5 mic. in diameter T. lanata

(3) Filaments 10-15 mic. in diameter T. distorta

(4) Filaments 12-17 mic. in diameter T. penicillata

2 Plants living in moist places r

(1) Filaments 10-15 mic. in diameter T. byssoidea

(2) Filaments 15-25 mic. in diameter T. ravenelii

II. Sheaths thick

1 Plants living in water

(1) Filaments 5-6 mic. in diameter T. setchellii

(2) Filaments 12-15 mic. in diameter T. limbata

2 Plants living in moist places; filaments 12-15 mic. in diameter

T. rupestris

Genus DESMONEMA

Plate XXIV, Fig. 64

Plant mass cespitose, penicillate; filaments somewhat dichotomously

divided, straight; sheaths thin; trichomes two or more within the sheath;

heterocysts basal; gonidia large, oval or elliptical, single or in short series;

wall of gonidium somewhat thick.

Plant mass 5-6 mm. in height, formed of penicillate fascicles; fila-

ments erect, repeatedly sub-dichotomously branched; trichomes 9-10 mic.

in diameter, constricted at the joints; cells three times shorter than the

diameter; heterocysts one, two or none D. wrangelii

Genus DIPLOCOLON

Plate XXIV, Fig. 65

Plant mass gelatinous, terrestrial; colonies irregular in shape, con-

stricted, somewhat club-shaped; filaments several, contorted within a

common tegument, branched; false branches solitary or in pairs, usually

arising between two heterocysts, but rarely in the immediate region of the

heterocysts; trichomes single within the sheath.

Colonies club-shaped, gelatinous, irregularly dilated, up to 1 mm. in

thickness, yellowish brown; filaments 20-28 mic. in diameter; trichomes

constricted at joints; cells and heterocysts 6-10 mic. in diameter, some-

what spherical D. heppii

SYNOPSIS OF THE BLUE-GREEN ALG#Z—MYXOPHYCE#® 247

Family IV. STIGONEMACEZ:

Filaments free, rarely laterally aggregated, scattered, frequently

branched; sheaths thick, firm, often irregular; trichomes consisting of one

or several rows of cells, with heterocysts; heterocysts often lateral, some-

times intercalary; reproduction by means of vegetative division, hormo-

gones and gonidia.

I. Sheaths distinct, definite

1 Filaments free

(1) Trichomes consisting of one row of cells

A. Branches of two kinds, the one cylindrical, the other flagel-

liform; heterocysts terminal or lateral

Mastigocoleus

Plate XXIV, Fig. 66

B Branches unilateral, usually tapering at the apex; heterocysts

intercalary Hapalosiphon

Plate XXV, Fig. 67

(2) Trichomes consisting of one to several rows of cells

A Branches unilateral, thin, finally forming hormogones

Fischerella

Figs. 68, 69

B Branches scattered; hormogones formed in the apices of the

branches or in special short branches

Stigonema

Fig. 70

2 Filaments growing together forming a cushion-like mass

Capsosira

Fig. 71

II. Sheaths confluent into a gelatinous amorphous mass

Nostochopsis

Plate XXVI,

Fig. 72

Genus MASTIGOCOLEUS

Plate XXIV, Fig. 66

Filaments free, irregularly branched; branches of two kinds, the one

cylindrical, the other flagelliform, tapering off into a hair-like apex;

sheaths continuous; trichomes, except in the branches, single within the

sheath; heterocysts single, rarely in pairs, terminal or lateral, sometimes

intercalary; gonidia unknown; reproduction by means of hormogones; cell

contents homogeneous.

Filaments 6-10 mic. in diameter; trichomes 3.5-6 mic. in diameter;

heterocysts 6-18 mic. wide and long; cell contents greenish

M. testarum

SYNOPSIS OF THE BLUE-GREEN ALGH—MYXOPHYCEZ 249

Genus HAPALOSIPHON

Plate XXV, Fig. 67

Plant mass cespitose-floccose, thin, aquatic; filaments free, not

growing together laterally, branched, consisting of a single row of cells,

rarely of two rows, enclosed within a sheath; branches erect, usually

about the same thickness as the creeping primary filament, commonly

unilateral, long, flexuous, very slightly tapering; sheaths continuous,

strong, of uniform thickness; sheaths of the branches thinner than those

of primary filaments, usually colorless; heterocysts intercalary; wall of

gonidium thick, yellowish brown.

I. Plants living in fresh water

1 Filaments decumbent, branched on all sides; branches 6-8 mic. in

diameter; cells elliptical depressed H. flexuosus

2 Plant mass cespitose, orange brown; primary filaments 11.5-12.5

mic. in diameter; trichomes 7.5-8 mic. in diameter

H. aureus

3 Plant mass floccose, czspitose, dull blue-green; primary filaments

21-24 mic. in diameter H. fontinalis

II. Plants living in hot water

1 Plant mass cushion-like, irregular or expanded, blue-green; prim-

ary filaments 3-6 mic. in diameter H. laminosus

2 Plant mass widely expanded, bright blue-green; trichomes 3-11

mic. in diameter H. major

III. Plants living on bark of trees

1 Plant mass cespitose, small, blue-green; filaments 4-7 mic. in diam-

eter H. intricatus

2 Filaments 7-10 mic. in diameter; trichomes 7-9.5 mic. in diameter

H. arboreus

Genus FISCHERELLA

Plate XXV, Figs. 68, 69

Plant mass forming a continuous, more or less expanded layer, ter-

restrial; filaments of two kinds; primary filaments creeping, containing

one or two rows of cells, unilaterally very much branched; branches or

secondary filaments erect, elongate, more slender than the primary, con-

taining very long hormogones within the sheath.

I. Plants living in moist places; primary filaments 6-9 mic. in diameter

F. ambigua

II. Plants living in moist places or in hot water; primary filaments 10-13

mic. in diameter F. thermalis

250 JOSEPHINE E. TILDEN

Genus STIGONEMA

Plate XXV, Fig. 70

Plants terrestrial or aquatic; plant mass rigid, blackish brown, or

cushion-like and soft; filaments free, rarely laterally aggregated, scat-

tered; trichomes, in the larger filaments, consisting of two or several rows

of cells; heterocysts often lateral, here and there intercalary, hormogones

developed in the apices i vegetative branches or in short special

branches.

I. Trichomes in the mature filaments consisting usually of a single row

of cells

1 Filaments 7-15 mic. in diameter; sheaths usually colorless

S. hormoides

2 Filaments 25 mic. in diameter; cells 14 mic. in diameter, 6-8 mic. in

length S. zrugineum

3 Filaments 24-26 mic. in diameter; sheaths thick, lamellose

S. panniforme

4 Filaments 14-38 mic. in diameter, sheaths yellowish brown

S. tomentosum

5 Filaments 24-45 mic. in diameter; sheaths thick, lamellose, color-

less or yellowish brown S. ocellatum

II. Trichomes in the mature filaments consisting usually of two or

several rows of cells

1 Filaments up to 35 mic. in diameter

(1) Filaments 18-28 mic. in diameter; cells often surrounded by a

special darker colored envelope S. minutum

(2) Filaments 27-37 mic. in diameter; cells throughout the entire

length of the filament uniformly divided

S. turfaceum

2 Filaments 40-90 mic. in diameter

(1) Hormogones 45 mic. in length, terminal, solitary or in series

S. informe

(2) Plants rigid; hormogones 45 mic. in length, lateral

S. mamillosum

Species not well understood S. brandegeei

Genus CAPSOSIRA

Plate XXV, Fig. 71

Plant mass hemispherical, cushion-like, attached by lower surface,

formed of filaments growing together laterally, aquatic; filaments erect,

branched, composed of a single row of cells; sheaths septate; heterocysts

SYNOPSIS OF THE BLUE-GREEN ALG2#—MYXOPHYCE% 251

intercalary and lateral; hormogones composed of from 10-20 cells; gonidia

spherical; wall of gonidium thick, brownish.

Filaments 7.5 mic. in diameter; cells 4-5 mic. in diameter; heterocysts

lateral C. brebissonii

Genus NOSTOCHOPSIS

Plate XXVI, Fig. 72

Plant mass or colony gelatinous, definite, aquatic; trichomes formed

of a single row of cells, branched; heterocysts intercalary and lateral,

pedicellate or sessile.

Colony, hollow; trichomes 4-9 mic. in diameter, 1 mm. in length;

branches unilateral, fastigiate, cylindrical below, torulose in upper por-

tions, somewhat club-shaped; heterocysts lateral, exserted, or intercalary

N. lobatus

Family V. RIVULARIACEZ

Filaments tapering from base to apex, ending in a colorless hair,

simple or branched; false branches due to development of new trichome

from a cell of the main trichome, usually occurring immediately under

an intercalary heterocyst—rarely by the perforation of the sheath between

two heterocysts by the trichome, as in Scytonema—either separating im-

mediately and forming a new sheath, or remaining for some time within

the original sheath; heterocysts usually present, usually basal, occasional-

ly intercalary; reproduction by means of vegetative division, hormogones

and gonidia.

I. Heterocysts not present Amphithrix

Plate XXVII

Fig. 73

II. Heterocysts present

1 Filaments free, simple or coalesced into a branched plant mass

(1) Sheaths cylindrical

A. Filaments simple or branched; false branches distinct, free

Calothrix

Fig. 74

B Filaments branched; false branches several (two to six) re-

maining within the original sheath or common tegument

Dichothrix

Fig. 75

C Filaments branched; false branches many (up to a hundred)

remaining within the original sheath or common tegu-

ment Polythrix

(2) Sheaths thick, saccate Sacconema

Plate XXVIII

Fig. 76

Pe ety S ath ec Gy Te Sea

ae ee

*« “ & ti os Gs fn

ope ne . : a a, = : <3 i

poe, pal ans Oe as

. aT se

a gpa

Sp abs

PLATE XXVI

SYNOPSIS OF THE BLUE-GREEN ALGE—MYXOPHYCEZ 253

2 Filaments coalesced into a crustaceous, spherical or hemispherical,

mucous or gelatinous plant mass or colony

(1) Heterocysts basal

A Filaments simple, parallel, associated in a crustaceous layer

Isactis

Plate XXIX, Fig. 77

B_ Filaments branched, radially arranged, associated in a spher-

ical or hemispherical colony Rivularia

Fig. 78

(2) Heterocysts intercalary Brachytrichia

Fig. 79

Genus AMPHITHRIX

Plate XXVII, Fig. 73

Plant mass crustaceous or cespitose, thin, expanded, of a purple

or violet color, consisting of two layers: the lower layer composed of

densely interwoven filaments or of minute, radiately disposed series of

cells; the upper layer consisting of simple erect filaments, closely packed

together and tapering to fine points; sheaths thin, close, continuous;

hormogones solitary or in series; heterocysts not present.

I. Plant mass crustaceous; filaments 1.5-2.2 mic. in diameter; 3-5

decimill. in length; cells equal to the diameter in length

A. janthina

II. Plant mass cespitose; filaments 2-3 mic. in diameter, 1-3 mm. in

length; cells shorter than their diameter A. violacea

Genus CALOTHRIX

Plate XXVII, Fig. 74

Plant mass consisting of penicillate tufts or a soft velvety expansion;

filaments simple or slightly branched; heterocysts basal or intercalary, ab-

sent in a few species; gonidia basal, seriate.

I. Heterocysts not present C. juliana

II. Heterocysts present

1 Plants living in salt water

(1) Heterocysts basal

A Plants fasciculate or penicillate, parasitic

a Filaments 12-15 mic. in diameter C. confervicola

b Filaments 21-29 mic. in diameter C. consociata

B Plants cespitose, often growing on rocks

a Filaments 8-12 mic. in diameter; cell contents violet

C. fusco-violacea

b Filaments 10-18 mic. in diameter; cell contents olive green

C. scopulorum

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PLATE XXVII

SYNOPSIS OF THE BLUE-GREEN ALG#—MYXOPHYCE/ 255

c Filaments 9-15 mic. in diameter; cell contents olive green

C. contarenii

d Filaments 15-20 mic. in diameter; cell contents olive green

C. pulvinata

e Plants parasitic; filaments 9-15 mic. in diameter, thickened

into a bulb at the base; cell contents blue-green

C. parasitica

(2) Heterocysts basal and intercalary

A Filaments 9-12 mic. in diameter, scarcely thickened at base

C, zruginea

B_ Filaments 15-18 mic. in diameter; false branches solitary

C. prolifera

C Filaments 12-21 mic. in diameter; false branches fasciculate at

the apex of the filament C. fasciculata

D Filaments 12-24 mic. in diameter; false branches in pairs, aris-

ing between two heterocysts C. vivipara

E Filaments 10-40 mic. in diameter, interwoven at base, decum-

bent C. pilosa

F Filaments 12-40 mic. in diameter; sheaths yellowish brown’

C. crustacea

2 Plants living in fresh water

(1) Plants epiphytic

A Filaments 5-7.5 mic. in diameter; trichomes 3.4-4 mic. in diam-

eter C. epiphytica

B Filaments 7-8 mic. in diameter; heterocysts basal, usually in

pairs C. scytonemicola

C Filaments 8-10 mic. in diameter; trichomes 6-9 mic. in diam-

eter, especially constricted at joints; heterocysts basal, in

pairs C. stagnalis

D Filaments 10-12 mic. in diameter, curved and bulbous-inflated

at the base; trichomes 7-8 mic. in diameter

C. fusca

E Filaments 5-15 mic. in diameter, sometimes thicker at the

base; trichomes 3.5-5.5 mic. in diameter

C. sandwicensis

F Filaments 15-16 mic. in diameter at base; sheaths thick, lamel-

lose, finally becoming brownish black; cells very short

C. breviarticulata

256 JOSEPHINE E. TILDEN

G Filaments 15-18 mic. in diameter; sheaths wide, often trun-

cate, almost colorless; trichomes 7-9 mic. in diameter

C. violacea

H Filaments 18-24 mic. in diameter; sheaths thick, gelatinous,

lamellose, finally ocreate C. adscendens

(2) Plants living in warm or hot water

A. Filaments 8-10 mic. in diameter; sheaths somewhat thick,

uniform, transparent, sometimes yellowish at base; heter-

ocysts basal, rarely intercalary C. thermalis

B Filaments 8-10 mic. in diameter; sheaths close, ocreate, trans-

parent, becoming yellowish brown; heterocysts basal and

intercalary, spherical or quadrate C. calida

C Filaments 10-11 mic. in diameter; sheaths close, thick, lamel-

lose, ocreate, transparent and yellowish

C. kuntzei

(3) Plants living on stones and wood

A. Filaments 9-10 mic. in diameter; sheaths narrow, close, uni-

form, colorless; trichomes 6-7 mic. in diameter

C. braunii

B Filaments 10-12 mic. in diameter; sheaths thick, close, uni-

form or ocreate, yellowish brown C. parietina

C Filaments 12-13 mic. in diameter; sheaths thin, close, uniform,

colorless or yellowish C. castellii

Genus DICHOTHRIX

Plate XXVII, Fig. 75

Plant mass cespitose, penicillate or pulvinate; filaments more or less

dichotomously branched; trichomes often several (two to six) enclosed

within the original sheath or common tegument; heterocysts sometimes

basal, sometimes intercalary, in one species not present.

I. Plants living in fresh water

1 Sheaths close, gradually tapering at the apex

(1) Plants living in hot water; filaments 15-25 mic. in diameter,

trichomes 5-6 mic. in diameter D. montana

(2) Filaments 10-12 mic. in diameter, flexuous, erect, radiating

D. orsiniana

(3) Plant mass encrusted with calcium carbonate; filaments 9-12.5

mic. in diameter, prostrate, not rigid D. calcarea

(4) Plants living in fresh or rarely salt water; filaments about 15

mic. in diameter; trichomes 5-9 mic. in diameter, constricted

at joints D. baueriana

SYNOPSIS OF THE BLUE-GREEN ALGE—MYXOPHYCEH 257

(5) Filaments 12-15 mic. in diameter; trichomes 10-15 mic. in

diameter, not constricted at joints D. olivacea

2 Sheaths lamellose, funnel-shaped at apex

(1) Filaments 9-12 mic. in diameter; trichomes 6 mic. in diameter

D. compacta

(2) Filaments 13 mic. in diameter; trichomes 6.5-7.5 mic. in diam-

eter D. meneghiniana

(3) Plant mass usually encrusted with calcium carbonate; fila-

ments 15-18 mic. in diameter; trichomes 6-8 mic. in diameter

D. gypsophila

(4) Filaments 25-28 mic. in diameter; trichomes 10-12 mic. in

diameter, bulbously inflated at the bases of the branches;

heterocysts light blue in color D. hosfordii

II. Plants living in salt water

1 Filaments 15-22 mic. in diameter, trichomes 7-9 mic. in diameter;

heterocysts basal D. rupicola

2 Filaments 20-30 mic. in diameter; trichomes 17-22 mic. in diam-

eter; heterocysts basal and intercalary D. fucicola

3 Filaments 25-35 mic. in diameter; trichomes 15 mic. in diameter;

heterocysts oblong, solitary D. penicillata

4 Filaments 22-30 mic. in diameter; trichomes 7.5-12.5 mic. in diam-

eter; heterocysts basal and intercalary D. utahensis

Genus POLYTHRIX

Plant mass filiform, branched, consisting of numerous filaments fas-

ciculately arranged, included within a common tegument; filaments dense-

ly crowded, branched; heterocysts terminal and intercalary.

Trichomes 5-6 mic. in diameter, tapering into a thin hair at the

apex; heterocysts somewhat spherical P. corymbosa

Genus SACCONEMA

Plate XXVIII, Fig. 76

Plant mass or colony small, gelatinous, lobed or torn; common tegu-

ment lamellose, very much folded and saccate, finally dissolved at apices,

containing from two to many trichomes; trichomes irregularly aggre-

gated, somewhat czspitose; false branches short, moniliform, not

coalesced; heterocysts basal, spherical; gonidia present.

Trichomes 8 mic. in diameter; heterocysts basal, spherical; gonidia

15 mic. in diameter; wall of gonidium roughened S. rupestre

mee i's

—~ Seer

A CY

~~ (#5) fi OY

XXVIII

260 JOSEPHINE E. TILDEN

Genus ISACTIS

Plate XXIX, Fig. 77 :

Plant mass flat, crustaceous, orbicular, thin, adhering by the lower

surface, growing at the margin; filaments parallel, erect, unbranched or

rarely sparingly branched; heterocysts basal; gonidia unknown.

I. Filaments decumbent at base; trichomes 7-9 mic. in diameter

I, plana ;

II. Filaments slightly swollen at base; trichomes 8-10 mic. in diameter

. I. centrifuga

Genus RIVULARIA

Plate XXIX, Fig. 78

Colonies spherical, hemispherical or inflated and lobed, solid or hol-

low, sometimes confluent into an indefinite mass; filaments radiating from

the center, repeatedly branched; sheaths conspicuous near the base of the

trichomes, near the periphery of the colony gelatinous and confluent;

heterocysts basal; gonidia more or less cylindrical and elongate, not

known in all species.

I. Filaments flagelliform, tapering towards the apex; gonidia present

1 Colonies hard; trichomes 4-7 mic. in diameter; gonidia 9-15 mic. in

diameter, especially cylindrical R. pisum

2 Colonies soft; trichomes 7-9 mic. in diameter; gonidia 10-18 mic. in

diameter, larger at the base R. natans

3 Colonies firm, solid, light green; gonidia cylindrical, frequently

curved, about nine times as long as broad R. incrustata

4 Colonies soft, solid; trichomes 8-10 mic. broad at the base; gonidia

8-18 mic. in diameter, cylindrical, straight or slightly curved

R. echinulata

II. Filaments gradually tapering; gonidia unknown

1 Colonies hollow when old

(1) Colonies soft; trichomes 4-5 mic. in diameter in lower portion,

somewhat constricted at joints R. polyotis

(2) Colonies soft; trichomes 2-5 mic. in diameter, cylindrical

R. nitida

2 Colonies solid

(1) Colonies not encrusted with calcium carbonate

A Plants living in fresh water

a Trichomes 4 mic. in diameter R. borealis

b Trichomes 6-10 mic. in diameter R. compacta

c Trichomes 9-12.5 mic. in diameter R. minutula

— oe,

SYNOPSIS OF THE BLUE-GREEN ALG#—MYXOPHYCEZ 261

d Trichomes continuous or indistinctly divided; heterocysts

10-12 mic. in diameter R, paradoxa

B Plants living in salt water; trichomes 2.5-5 mic. in diameter

R. atra

(2) Colonies encrusted with calcium carbonate

A Colonies hemispherical, finally confluent and forming a hard,

stony crust; trichomes 4-7.5 mic. in diameter

R. hematites

B Colonies small, somewhat hard; trichomes 4-9 mic. in diam-

eter R, dura

C Colonies at first hemispherical, afterwards forming a gelatin-

ous crust, indurated with calcium carbonate in the interior;

trichomes 5-9 mic. in diameter R. coadunata

D Colonies spherical, hard; trichomes 4-16 mic. in diameter

R. bornetiana

Genus BRACHYTRICHIA

Plate XXIX, Fig. 79

Colonies at first solid, finally becoming hollow, made up of Nostoc-

like filaments embedded in gelatin; filaments flexuously curved, parallel,

above tapering and drawn out into a hair at the apex, very much

branched; sheaths distinct in the young filaments, tubular, finally becom-

ing confluent and invisible; heterocysts intercalary, arranged without any

order.

Colonies up to 5 cm. in diameter, plicate-expanded and bullate, con-

fluent, blackish green. B. quoyi

Department of Botany,

University of Minnesota.

GLOSSARY

The proper terms to be used in connection with the blue-green

alge have not yet become definitely established. The terms and defin-

itions given in this synopsis are merely provisional, in case better ones

can be found. Some difficulty has been experienced with the terms:

“plant mass,” “colony,” “family,” “thallus,” etc. The definition of each,

as the author conceives the meaning, is given in the glossary.

Plant mass, the usually shapeless mass of individual plants remaining in

close proximity to each other after their formation, either because

nothing occurs to separate them or because they are definitely held

together by a gelatinous excretion

Colony, a mass of plants of more or less definite shape, large enough to

be detected by the naked eye

262

JOSEPHINE E. TILDEN

Family, a mass of plants of microscopic size and somewhat definite

shape, quite evidently arising from the division of a single cell

Plant, in the coccoGoNE# a single cell; in the HORMOGONEZ a single tri-

chome. In the latter case it may be thought better by some to

consider “plant” and “filament” as synonymous terms.

Adherent, clinging to, or united

with

Adnate, touching closely or

broadly

Agglutinated, glued together

Aggregated, forming a mass or col-

lection, but not cohering

Amorphous, structureless

Anastomose, to run together in a

netlike manner

Angular, having angles; sharp cor-

nered

Apex, the end opposite the point of

attachment; tip

Appressed, pressed closely against

Approximate, near, about

Aquatic, living in water

Arachnoid, cobwebby

Articulate, jointed with cells

Asexual, without sex

Base, the point of attachment

Brackish, somewhat salty

Bulbous, with a bulb

Bullate, swollen

Bullose, swollen

Czspitose, in tufts or dense

bunches

Calcareous, composed of or con-

taining lime

Calyptra, a cap or lid

Capitate, furnished with a globose

head

Carneous, fleshy

Cartilaginous, firm and tough like

cartilage

Catenate, joined in a continuous se-

ries; in a chain

Cell, a closed sac, surrounded by a

wall of cellulose, containing pro-

_toplasm and a single nucleus

Cell sap, the watery fluid of a cell

which separates from the proto-

plasm as one or more vacuoles

Cell wall, the membrane enclosing

the cell contents

Cellulose, the cell wall substance of

plants

Centrifugally, from the center

Centripetally, toward the center

Chlorophyll, the green coloring

matter contained in plants; leaf-

green

Chromatophore, a plastid, contain-

ing a coloring matter

Cilium (pl. cilia), one of the vibra-

tile, protoplasmic processes

which serves to propel zoogonid-

ia through the water

Circinate, rolled from the end

Clathrate, with openings like lat-

tice work

Clavate, club-shaped

Coalesced, grown together, united

Coalescence, the complete union of

similar things

Collateral, side by side, secondary

Colony, a group of independent

cells surrounded by a common

investment; a mass of plants of

more or less definite shape, large

enough to be detected by the

naked eye

Concentric, with a common center

Confluent, growing or running to-

gether

Conidium, gonidium; a gonidium

which is abstricted from the apex

of a stalk

Constricted, narrowed

places

Contiguous, near or in contact

Contorted, twisted

Contractile, able to contract

Convolute, rolled together

Coriacious, leathery, tough

Crenate, wavy

Crisped, curled

Crustaceous, crust-like

Cuspidate, pointed, with a tooth

Decumbent, lying down

Deliquescent, dissolving

Dense, crowded together

Depressed-globose, globular, with

the poles slightly flattened

Dichotomous, two-forked; furcate

Dichotomy, division into two

branches

Diffluent, dissolving .

Disc, any flat circular area

in certain

SYNOPSIS OF THE BLUE-GREEN ALGA7E—MYXOPHYCE

Disc-shaped, flat and circular

Dissepiment, cross wall

Distal, pertaining to the apex

Divaricate, spreading

Diverging, separating

Eccentric, without a common cen-

ter

Elongate, lengthened, very long

Endophyte, a plant living within

another organism, usually as a

parasite

Entire, not toothed

Epiphyte, a plant growing upon the

outside of another plant, but not

nourished by it

Equilateral, with equal sides

Family, a mass of plants of micro-

scopic size and somewhat def-

inite shape quite evidently aris-

ing from the division of a single

cell

Fascicle, bundle

Fasciculate, in bundles

Fastigiate, tapering to a point

Fenestrate, window-like

Fibrille, little threads

Fibrillose, made up of small fibers

Fibrous, of fibers

Filament, the trichome together

with its sheath; a fine thread

Filamentous, thread-like, composed

of filaments

Filiform, thread-shaped

Fission, splitting; cell division in

which cell separates into two

nearly equal portions, especially

as a mode of reproduction

Flaccid, soft, flabby

Flagelliform, whip-like

Flexuous, flexible

Floccose, composed of matted,

woolly hairs

Flocculent, woolly

Foliaceous, leaf-like

Foliose, leaf-like

Gelatinous, jelly-like

Geminate, paired

Geniculate, bent abruptly like a

bent knee

Genuflexuous, bent abruptly

Glaucous, sea-green, gray-green

Globose, like a ball

Globular, spherical or nearly so

Gonidangium, the cell in which go-

nidia are produced

263

Gonidium, a reproductive cell de-

veloped asexually; a specialized

reproductive cell capable by it-

self of giving rise to a new or-

ganism

Granular, with granules

Granule, a small grain

Granulose, with granules

Gregarious, growing in association,

but not matted together

Grumose, grumous, like a cluster

of grains

Habit, the general appearance or

characteristic manner of growth

of a plant

Habitat, the locality or region, or

the kind of situation in which a

plant is naturally found

Heterocyst, a cell uniformly larger

than its neighbors, but of doubt-

ful function

Hirsute, with coarse hairs

Homogeneous, uniform in charac-

ter or substance

Hormogone, a short chain of cells

broken off from a mature plant,

by means of which vegetative

multiplication is effected

Host, a plant which supports a

parasite (or an epiphyte?)

Hyaline, clear and colorless, trans-

parent

Immersed, sunken below the sur-

face

Impregnated, filled with

Indurated, hardened

Inequilateral, with unequal sides

Inflated, swollen

Integument, any outer covering

Intercalary, inserted between

Intricate, tangled, involved

Inundated, flooded

Investment, a covering

Laciniate, torn

Lacunose, hollowed

Lamelliform, plate-like

Lamellose, with plates or blades

Lenticular, lens-shaped

Lubricous, slippery, slimy

Lumen, cavity

Mammillate, mammillose, with nip-

ple-like projections

Mammilliform, nipple-like

Marginal, at the edge

Membranaceous, papery

Moniliform, chain-like

264

Motile, able to.move

Mucilaginous, jelly-like

Multicellular, of several to many

cells

Nodule, a little knot or lump

Nucleus, a differentiated round or

oval body embedded in the pro-

toplasm of a cell

Obovate, ovate, but with the point

of attachment at the lower end

Ocrea, a sheath

Ocreated, sheathed

Orbicular, circular

Oval, elliptical

Ovoid, egg-shaped

Pannose, ragged

Papillose, with a little point or nip-

ple

Parasite, a plant that lives on or in

some other organism from which

it derives its nourishment for the

whole or a part of its existence

Parenchyma, the soft, thin-walled

cellular tissue of plants

Pedicel, a small or delicate sup-

porting stalk

Pedicellate, stalked

Pellucid, clear

Penicillate, like a brush

Periphery, edge

Phycocyanin, a blue pigment con-

tained in the chromatophores of

the blue-green alge

Pigment spot, a specialized mass of

cytoplasm permeated by a red

coloring matter, present in the

motile cells of many algx; eye-

spot ;

Piliferous, bearing hairs

Pilose, hairy

Plant, in the COccoGoNEz a single

cell; in the HORMOGONEZ a single

trichome

Plant mass, the usually shapeless

mass of individual plants remain-

ing in close proximity to each

other after their formation,

either because nothing occurs to

separate them or because they

are definitely held together by a

gelatinous excretion

Plicate, folded or ridged

Polar, at the end

Polygonal, many-sided

Polyhedral, many-angled

JOSEPHINE E. TILDEN

Polymorphous, of many forms

Proliferated, grown out

Protoplasm, the viscid, contractile,

semiliquid, more or less granular,

substance that forms the princi-

pal portion of an animal or vege-

table cell

Prostrate, flat, lying down

Pseudo-parenchymatous, like par-

enchyma

Pubescent, finely hairy

Pulverulent, powdery

Pulvinate, cushion-like

Punctate, dotted

Punctiform, dot-like

Pustular, like a swelling

Pyrenoid, a small colorless mass of

proteid substance seen in many

alga, which may be regarded as

reserve material

Quadrate, square, in fours

Radial, pertaining to a radius, as of

a circle or sphere

Rectilinear, straight

Refractive, refringent, bending or

turning aside as a light ray

Reniform, kidney-shaped

Reproduction, the development of

one or more new organisms from

the whole or from a part of the

protoplasm of a parent organism

Rotund, round

Rugose, furrowed, roughened

Saccate, sack-like

Segment, one of the parts into

which an object is naturally di-

vided

Septate, divided by partitions

Seriate, in a row

Sessile, without a stalk

Sheath, a gelatinous, usually tubu-

lar, envelope surrounding a plant

Silicious, containing silica

Sinuate, snake-like, twisted

Sinus, a gulf or indentation

Spatulate, shaped like a spoon

Spherical, ball-like

Spongiose, spongy

Stellate, star-like

Stratified, in layers

Stratum, a layer

Striated, having fine markings

Sub, slightly, somewhat

Submerged, sunken

Substratum, surface on which the

plant grows

SYNOPSIS OF THE BLUE-GREEN ALG2—MYXOPHYCE/E

Superposed, placed one above an-

other

Tegument, covering

Tenacious, firm, tough

Terebriform, screw-like

Terminal, end

Terrestrial, growing on the ground

Thallus, a plant-body without true

root, stem or leaf; used incor-

rectly instead of “plant mass”

Tomentose, closely hairy

Tortuous, twisted

Torulose, chain-like

Trichome, the entire number of

cells of a multicellular plant, not

including the sheath

Truncate, cut off abruptly

265

Tuberculate, tuberculose, warted

Tubular, tube-like

Ultimate, last end

Uncinate, hooked at the end

Undulate, wavy

Unicellular, one-celled

Unilateral, one-sided

Vacuole, a cavity containing a wat-

ny fluid in the protoplasm of a

Ventricose, a swelling out on one

side or in the middle

Verrucose, warted

Vesicle, a small bladder-like cavity

Vesicular, bladdery

Villous, long hairy

Zonate, disposed in the form of

zones

BIBLIOGRAPHY

Fritscu, F. E.

Studies on Cyanophycee.

New Phyt. 3:85-96. f. 76.

Beihefte Bot. Centralbl. 18’:

Ap 1904; III.

194-

Cytological Studies in Cyanophycee. Univ. Calif. Pub. Bot. 2 :237-296.

32216, pl. 10... D 91904: TL,

21a pl. Fae 1905

GARDNER, N. L.

pl. 21-26. N 1906

HARSHBERGER, J. W.

Algal stalactites in Bermuda.

Murray, G.

Torreya 14:195-197.

O 1914

Calcareous Pebbles formed by Algze. Phycological Memoirs, London.

April 1895

NE son, N. P. B.

Observations upon some Algze which cause “Water Bloom”.

f. 1-3. Mr 1903

Stud. 3:51-56. pl. 14.

Puiurps, O. P.

Minn. Bot.

A comparative study of the Cytology and Movements of the Cyanophy-

cee. Trans. and Proc. Bot. Soc. Pa. 1:237-335. pl. 23-25.

Roppy, H. J.

1904

Concretions in streams formed by the agency of Bluegreen Alge and

related Plants.

Proc. Am. Philos, Soc. 54:246-258. f.1, 2. Au 1915

266 JOSEPHINE E. TILDEN

Spratt, E. R.

Some Observations in the Life-History of Anabena Cycadee. Ann. Bot.

25 :369-380. pl. 32. Ap 1911

TiLpEN, J. E.

Some new species of Minnesota Alge which live in a Calcareous or

Siliceous Matrix. Bot. Gaz. 23:95-104. pl. 7-9. F 1897

Trven, J. E.

On some Algal Stalactites of the Yellowstone National Park. Bot. Gaz.

24 :194-199. pl. 8. S 1897

TILpEN, J. E.

Observations on some West American Thermal Alge. Bot. Gas. 25 :89-

105. pl. 8-10. F 1898

TiLpEN, J. E.

The Myxophycee of North America and adjacent regions, including

Central America, Greenland, Bermuda, the West Indies and Ha-

waii. Minnesota Alge. 1:1-328. 1910

TILpEN, J. E.

Index Algarum Universalis. 1915-1917

WEED, W. H.

The Formation of Travertine and Siliceous Sinter by the Vegetation of

Hot Springs. U. S. Geol. Survey. Ann. Report ix. 1887-8.

West, G. S

Alge. I. 1916. (Myxophycez. 1-48)

DEPARTMENT OF NOTES, REVIEWS, ETC.

It is the purpose, in this department, to present from time to time brief original

notes, both of methods of work and of results, by members of the Society. All

members are invited to submit such items. In addition to these there will be given a

few brief abstracts of recent work of more general interest to students and teachers.

There will be no attempt to make these abstracts exhaustive. They will illustrate progress

without attempting to define it, and will thus give to the teacher current illustrations, and

to the isolated student suggestions of suitable fields of investigation.—[Editor.]

TECHNICAL METHODS

1. Electrifying the Microtome

As this laboratory is specializing on a collection of serial sections

of pig embryos, it became necessary in some manner to reduce the

amount of time consumed as much as possible. After a careful

study of the problem, it was decided to attach an electric motor to the

microtome, thus leaving both hands of the technician free to handle

the ribbons as they came off the knife. In this manner over one-

third of the time ordinarily spent in section cutting was saved.

As the microtome used was an old type, of the steam engine

crank motion with a large counter balanced fly wheel, it was rather

hard to find the right kind of motor, especially as the fly wheel was

not grooved for a belt, and could not be as its walls were too thin.

Also there was no cheap and handy controller for ordinary motors to

be had, except a belt tightener such as usually supplied on factory

electrified microtomes.

Seeing the Western Electric sewing machine motor advertised,

I asked the agent for a demonstration, and since then our motor

trouble has been solved. The device consists of a small motor with

a cork friction wheel, and mounted on a base so that when put under-

neath a sewing machine fly wheel a strong spring in the base will

press the cork friction wheel against the fly wheel of the sewing

machine. The motor was turned on its side with its base bolted to a

block, and that block bolted to the table. The microtome wheel was

then backed up onto the cork friction wheel of the motor until the

requisite tension was secured, and then it also was bolted to the table.

Thus both motor and microtome are both rigidly fastened down

(Plate XXX).

Proceedings from the Zoological Laboratory of the North Dakota Agricultural Col-

lege, No. 3.

268

NOTES, REVIEWS, ETC.

TTA

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PLATE XXX

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AMERICAN MICROSCOPICAL SOCIETY 269

Variations in speed are obtained by means of the excellent foot

controller which accompanies the motor. Although this controller

allows only six variations in speed, I have always been able to regu-

late with complete satisfaction. While cutting serial sections my

assistant never has to touch the fly wheel with his hands, as he is

able merely by using the foot control to move the paraffin block a

fraction of an inch at a time, as well as being able to cut the sections

one by one if necessary. In this manner I have been able to cut

celloidin sections with a slanting knife.

The outfit comes complete with motor, foot controller, and

connecting cord, from the Western Electric Company or any of its

agents, for $15. By its use I have already saved its price in the

amount of time saved by my assistant, who is paid by the hour.

2. Chart Making

At best, chart making is laborious, and if the maker is not

somewhat versed in the art of freehand drawing, the results are not

all that could be wished. Especially is this true when the charts are

copied from text book illustrations by students or student assistants.

To obviate this difficulty I have provided this laboratory with

a “pantograph,” an ingenious arrangement of levers costing $1.00,

by which illustration and drawings may be copied exactly up to a

magnification of eight diameters, which is usually all that is desired.

If, however, a higher magnification is necessary, the drawing may

be remagnified with the pantograph to any degree wanted. One

merely provides a pencil on the end of one of the levers, sets it for

the required magnification, and then traces over the drawing which

is to be copied with a pointer, and the enlarged drawing is made by

the pencil. In this way, and by using gummed black letters for label-

ing, charts, perfect in all respects, may be obtained by assistants who

may never have had any artistic training.

It was found that charts hanging on the wall in the laboratory

were attacked by the numerous “silver fish” (Lepisma saccharina),

and the ink and colors eaten off. In the endeavor to stop this, every

chart was sprayed with shellac, but as this did not entirely abate the

nuisance, arsenic in small quantities was mixed with the inks and

paints. At least the silver fish will have but one meal.

270 NOTES, REVIEWS, ETC.

3. Histological Technic

All tissues while dehydrating are kept in a rubber noppered

bottle connected with a water jet vacuum pump. Thus while the

tissues are dehydrating they are also being freed of air bubbles.

Under a rather low vacuum it is surprising the amount of air which

can be taken from most tissues.

In imbedding ordinary small blocks of tissue, benzine is used in

place of xylol or benzole. In fact I have been using benzine for

imbedding, dissolving paraffin off sections, cleaning slides, etc., where

xylol was formerly used. In this way the cost of imbedding and

section making is greatly reduced, as xylol is so very high in price

at the present time. Xylol is used only in clearing sections before

being mounted in xylol-balsam.

As a large number of horse tissues are sectioned and mounted

each year for my class in veterinary histology, I have obtained very

good results by using the paraffin method rather than the usual

celloidin procedure. As these tissues are very tough, careful technic

is necessary with paraffin, but the results, ie., clearer, thinner sec-

tions, and greater ease of handling, fully repay the added trouble.

These tough tissues, after dehydration and a stay in absolute

alcohol, are then placed in a beaker in which there are two separate

layers, one of cedar oil on the bottom, and one of absolute alcohol on

top. The block is then placed in the beaker in the absolute, resting

on the surface of the cedar oil. Gradually the block becomes infil-

trated with cedar oil, sinking further and further into it. When the

block comes to rest upon the bottom of the beaker it is then fully

cleared and ready to be infiltrated with low melting point paraffin.

Finally it is imbedded in hard paraffin, and sectioned with a slanting

knife, keeping the sections from curling with a camels hair brush or

a needle.

The paraffin used is the commercial PAROWAX, its melting

point lowered if necessary by adding liquid petrolatum, or raised by

adding olive oil. This is fully as efficient as the higher priced im-

bedding paraffins.

I have obtained an alcohol which approaches absolute alcohol

for most practical imbedding purposes by distilling 95% alcohol, and

AMERICAN MICROSCOPICAL SOCIETY 271

storing the distillate in bottles, the bottoms of which are covered with

a layer of anhydrous copper sulfate.

HERBERT EpMonp METCALF.

North Dakota

Agricultural College.

FuRTHER NOTES ON THE REARING OF VOLVOX

In the April number of these Transactions for the year 1916, I

related some experiences in the rearing of Volvox aureus. Since the

two cultures whose histories were there given have ceased to yield

and have been destroyed, the concluding chapter concerning those

cultures may now be written. A summary of their histories from

the time of writing the earlier account is as follows:

Culture No. 1, collected Oct. 27, 1915, on Apr. 27, 1916, con-

tained many thousands of Volvox colonies. Then followed a period

of decline until on June 7 not over six colonies were seen. During

June there was a rapid increase. On June 17 a few colonies were

seen. On June 19 over a hundred were counted and on July 10

they were very numerous. This was followed by a decline, for on

Aug. 9 only about one hundred colonies were noted, but on Aug.

17 these had increased to more than a thousand, and on Sept. 7 the

number was estimated at three to five thousand. At the time of

the next observation, Oct. 13, very few colonies were present, but

desmids had become very numerous. Thinking that the cool tem-

perature outside might stimulate the Volvox to grow, the culture was

put outside the window on Oct. 16. Eight days later there were sev-

eral hundred colonies. Ice was found on the top of the water of the

culture on the 16th of Nov., when the culture was brought inside.

On the following day a few colonies were seen. Examinations were

made on the following dates but no colonies could be found: Jan.

18, 1917, Feb. 16, Mar. 26, June 21. The culture was found with

blue-greens on the last date and was destroyed.

In this culture to which no fresh water, no organic matter and

no salts had been added since its establishment, Volvox grew for a

little over a year and at times even thrived. Toward the close of

the period the amount of available food materials must have been

very slight.

272 NOTES, REVIEWS, ETC.

Culture No. 2 established at the same time as No. 1, and kept

under like conditions, ran a somewhat similar course, but died out

earlier since no colonies were seen after Aug. 9, 1916. Apr. 20,

1916, several thousand colonies were seen and on May 4 they were

even more numerous. On May 16 they were on the decline, but on

May 22, June 17 and June 30 the record states that there were sev-

eral thousand colonies present. On July 10 three or four thousand

colonies were seen. These had all disappeared by Aug. 9, and no

more were seen on the following dates of examination: Sept. 9,

Oct. 18, Nov. 16, Jan. 18, 1917, Feb. 16, Mar. 26, June 21, 1917.

The culture which now was foul with blue-greens was destroyed on

the last date.

A number of cultures of Volvox globator were started July 12,

1916, and kept under the same conditions as the preceding cultures.

The colonies soon disappeared, however, and none have since ap-

peared. The cause of their disappearance is not even conjectured.

Zoological Laboratory, GerorcE R. LARUE.

University of Michigan.

PyRIDINE SILVER FOR BONE SECTIONS

As the different walls of a bone véry often show unlike differ-

entiations, an entire section should be prepared and not a small

piece from one wall.

Grind one surface until smooth and perfectly flat. Fasten to a

clean slide in hard balsam. Grind to microscopic thinness and polish.

Make a 1% solution of silver nitrate crystals in pyridine, in which

the crystals are soluble. Place slide and section in the solution and

let stand over night. During the night the section will probably be |

released from the slide. Clean and dry the section. Mount on a

clean slide in hard balsam with cover glass. If the preparation is

made by daylight, keep the silver solution in a dark place. By this

method I have made beautiful sections of those I had earlier dis-

carded as worthless.

J. S. Foote.

AMERICAN MICROSCOPICAL SOCIETY 273

ENTOMOLOGICAL NOTES

Parthenogenesis in Aleurodes.—Williams (717, Journ. Genetics,

6 :256-267), in a paper entitled “Some Problems of Sex Ratios and

Parthenogenesis,” presents results of a study of sex relations in a

common and widely distributed white-fly, Aleurodes vaporariorum,

which occurs in North and South America, the West Indies, Europe,

and New Zealand. This insect has two races, one in England which

produces females parthenogenetically, and the other in the United

States produces males parthenogenetically. Fertilized eggs yield an

equality of the sexes, and since the eggs would produce one sex

only if unfertilized, it is argued that there must be two kinds of

sperm, both of which develop in equal numbers. The similarity to

the case of Phylloxera caryecaulis is indicated, the chief differences

being the production of both sexes in sexual families and the ab-

sence of regular alternation of sexual and parthenogenetic genera-

tions. The occurrence of “male-” and “female-producing” races

of a single species, each of different geographical distribution and

varying sex proportions, raises the question of their origin. “Ex-

ternal conditions may alter the sex ratio in a colony by preventing

pairing.” The effect of partial or complete failure to pair on the

sex ratio in both races is discussed. ‘‘Male-producing” species or

races are more liable to die out, while in the “female-producing

races, the male sex is gradually lost, the race becoming in time com-

pletely parthenogenetic.” The original home of a species may be

indicated by the presence of both sexes, in case it is represented in

another locality by only one sex. An illustration is drawn from

the Pear Thrips of California, Teniothrips inconsequens, which

occurs in the United States, Canada, and Europe. In America,

males are not known, but in Europe both sexes are common. This

is regarded as evidence that the original home of the species was

in Europe and that females carried to Amreica have continued to

reproduce parthenogenetically. “Maleness” would be introduced if

a male be imported and subsequent mating with a female accom-

plished. Males might then become quite general. “The resulting

events would, however, depend on the sexes produced by fertilized

eggs and the relative fecundity of fertilized and parthenogenetic

females.’”’ Another example closely analogous seems to appear in

274 NOTES, REVIEWS, ETC.

a second species of thrips, olothrips fasciatus, common in England

and represented usually by males and females, the latter exceeding

the former slightly in number. This same species is also rather

common in the United States but, of the large number of individ-

uals studied from different localities, only two males have thus

far been recorded. Mention is also made of the chalcid, Tricho-

gramma pretiosa, which occurs in two morphologically indistin-

guishable races, one in America and the other in Europe. The former

produces males only by parthenogenesis, while the latter gives

females only or a mixture of males and females.

Sex in Anthothrips—Shull (17, Genetics, 2:480-488), in a

study of “Sex Determination in Anthothrips verbasci,”’ finds that

males and females are abundant, that fully formed spermatozoa

are produced by the male, and that copulation is of frequent occur-

rence in nature. Furthermore, virgin females also deposit unfer-

tilized eggs and the results of experiments show that such eggs

produce only individuals of the male sex. Females, which were

known to have mated either in captivity or in nature, were isolated

and the immediate progeny examined. In some cases, both sons

and daughters were produced; in others, only sons appeared, the

interpretation of the latter being the development from unfertilized

eggs even though the mother had been fertilized. Evidence is pre-

sented to support the conclusion that the fertilized eggs produce

only females. This case is then a parallel to that of the honey-

bee, and it seems that there is “some means of withholding the

spermatozoa, so that unfertilized eggs may be laid even while sperma-

tozoa are present.”

Amitosis in Adipose Cells—Nakahara (’17, Anat. Rec., 13:81-

86) finds that the adipose cells of various insects furnish “good evi-

dence that amitosis does not mean the approach of degeneration,

or aberration at all, but this kind of nuclear division may be chiefly,

if not entirely, to secure the increase of the nuclear surface to meet

the physiological necessity which is due to active metobolic inter-

changes between nucleus and cytoplasm.” Beginning at an early

stage, the adipose cell nucleus continues to divide amitotically, while

late in the third larval stage the cell stores up albuminous granules

in the cell-body. Thus the cell-nuclei undergoing amitosis have not

AMERICAN MICROSCOPICAL SOCIETY 275

degenerated but the cells proceed with their usual active processes.

Furthermore, it is claimed that the nuclei ‘themselves take a direct

part in the formation of the albuminous granules.”’

Nucleoli of Silk-gland Cells—Nakahara (’17, Journ. Morph.,

29:55-73) has made a morphological and chemical study of the

nucleoli of the silk-gland cells in the larve of a butterfly, Pieris

rape, and those of a caddis-fly, Neuronia postica. These cells are

characterized by multiple nucleoli, and while nucleoli are now gen-

erally considered as passive by-products of nuclear activity, their

role in the process of secretion has not been fully established. In

the insects mentioned, the nucleoli increase in number by the division

of the preexisting ones and increase quantitatively as the gland

becomes more functional. Some of the nucleoli migrate into the

cell-body, this migration continuing throughout the life of the func-

tional cell, and apparently constitute at least one source of the

secretion product of the cell. Migrating nucleoli appear to “give

off phosphorus to form themselves one of the lowest members of

the nuclein series.”

Origin of Photogenic Organs.—Hess (17, Ent. News, 28 :304-

310) has studied the “Origin and Development of the Photogenic

Organs of Photuris pennsylvanica De Geer (Col.).” Three con-

flicting views exist concerning the origin of these organs: (1) that

they are modified hypodermal cells, (2) that they are formed from

both ectoderm and mesoderm, and (3) that they originate from fat

cells. Recent papers by Vogel (1912) and Williams (1916) present

evidence in support of the view that the light producing organ arises

from fat cells. Hess finds that in 14-day embryos the hypodermis

on the ventro-lateral portion of each side of the eighth abdominal

segment shows a proliferation and enlargement of the cells. As

development goes on, these regions appear successively as a distinct

nodule on the inner surface of the hypodermis ; as organs completely

separated from the hypodermis except at the two ends of each

where they remain attached; and as distinct, two-layered organs

supplied with tracheal and nerve connections. It thus appears that

“instead of arising from fat cells, the embryonic organ is formed by

a proliferation of hypodermal cells, which are ectodermal ia origin.”

276 NOTES, REVIEWS, ETC.

Hibernation.—Baumberger (17, Annals Ent. Soc. Am., 10:179-

186) has studied certain insects with reference to the phenomenon

of hibernation, inspecting, among other features, the commonly con-

sidered hypothesis that low temperature or low mean temperature

is conducive to its appearance. Experiments with the banana fly,

Drosophila melanogaster, showed that in no case was a persistent

quiescent condition developed by the low temperatures. “Larve

and adults after twenty days in ice box were immediately activated

by high temperature.” Humidity experiments with the larve of

Isia isabella and Apantesis nais, which hibernate at a definite date

irrespective of low temperatures, showed that “high temperature,

abundant food and any relative humidity is not sufficient stimulus

to overcome the ‘tendency’ of the insects to hibernate.” It is there-

fore concluded that this quiesence is predetermined and in those

insects having a definite periodical hibernation, the “quiescence can

only be overcome by a certain period of low temperature and the

organism then by a compensatory lengthening of the next stage

regains its normal rhythm. Other insects are more plastic and

instead of showing a definite period of hibernation, merely remain

quiescent during periods of low temperature, and are active imme-

diately after the temperature is raised.” The latter class can be

reared continuously in the greenhouse. Every degree of this peri-

odicity is represented among insects. It is concluded that insects

hibernate: (1) as adults when the food habits permit the proper

oviposition at the earliest warm weather, (2) as larve when a

protection from low temperature permits feeding at the latest date

possible, and (3) as pupe or eggs “because they are nonfeeding

resistant stages.’’ It appears from the evidence that the phenom-

enon of hibernation is the result of repeated effect of winter, and

that the rhythmic character has been determined by the habits of

the species.

Nature of the Cervical Region—Crampton (717, Annals Ent.

Soc. Am., 10:187-197) presents a critical discussion and review of

the evidence bearing on the true nature of the neck region of insects

(Veracervix; Cervicum; “Microthorax”). The presence of plates

in the neck region has led to the development in the past of theories

(1) that the cervicum represents a “labial segment,” or (2) that it

AMERICAN MICROSCOPICAL SOCIETY 277

is a vestigial segment of the thorax. Crampton assembles the evi-

dence to show that the neck is in every way homologous with the

other intersegmental areas between the thoracic segments and does

not represent a segment at all; that this region, like other inter-

segmental ones, has no ganglia or other segmental structures, either

in the adult or embryonic stages; that the labium is not its append-

age; and that a labial segment already exists in the head to which

the labium is attached. While intersegmental plates between the

other thoracic segments have not been preserved in many of the

Pterygotan insects, and while only traces of them appear in still

others, yet they do appear in the Apterygotan insects, such as Japy+

and Eosentomon, and “since these are among the most primitive

of living insects, we are justified in assuming that the conditions

which they present approximate the original ones, in many respects.”

Inanition of Dermestid Larve.—Wodsedalek (17,) Science,

46 :366-367) reports remarkable viability in the larve of Tvogo-

derma tarsale (Dermestide). In starvation experiments, larve

lived without food as follows: newly hatched, four months; one-

fourth grown larve, about fourteen months; those about one-half

grown, almost three years; those three-fourths grown, four years;

most of the largest specimens, over four years, several over four

and one-half years; and one of the largest, five years and approxi-

mately two months. Gradual reduction in body-size was one char-

acteristic of the starvation effect. Ecdysis also occurred at inter-

vals, the successive cast skins being smaller and smaller. Starved

individuals which had almost attained the maximum decrease again

grew in size when supplied with plenty of food, alternate inanition

and feeding accompanied by alternate reduction and increase of size

was aecomplished three distinct times with the same individuals.

Development of Malpighian Tubules—Nelson (717, Science,

46 :343-345) contributes to the knowledge of the relation of the

Malpighian tubules to the hind-intestine in the larve of the honey-

bee. The mid-intestine is essentially a blind sac and subsequently

there is established a connection between the mid-intestine and the

hind-intestine. In the feeding larva of the honey-bee, the caudal

ends of the tubules are blind from the time of hatching up to the

sealing of the cell. After the cell is capped, a communication be-

278 NOTES, REVIEWS, ETC.

tween the mid- and hind-intestine is developed through which the

fecal accumulations of the former are expelled and, at the same

time, each of the four Malpighian tubules establishes connection

with the hind-intestine by a fine canal. ‘The history of the Mal- »

pighian tubules and that of the mid-intestine during the feeding

period of larval life are therefore parallel in that both, in addition

to performing their original functions, retain and store up the

accumulated excreta which is discharged only after feeding ceases,

when such discharge on the interior of the cell occupied by the larva

would not involve contamination of the food.”

Starvation and Wing Development.—Gregory (’17, Biol. Bull.,

33 :296-303), in a study of the effect of starvation on the wing devel-

opment of the green pea aphid, Microsiphum destructor, offers data

leading to the conclusion that external conditions control the appear-

ance of wings in this species. Under normal conditions, winged and

wingless individuals appear in varying numbers at different periods

in the life history, the wingless forms predominating in the early

part of the season, while the winged forms multiply toward the end

of the summer. The summary of all experiments on wingless par-

ents shows “that 25 starved individuals produced 1,257 young, 46

per cent. of which had wings; that 22 normal individuals produced

1,149 young, 9.7 per cent. of which were winged. The starved

mothers had about three times as many winged young as the con-

trol.” Since the food supply was the only variable factor, it alone

must be responsible for the results. Temperature may have an

indirect effect in decreasing the appearance of wings, and there is

some possibility that overcrowding on the plants out in nature may

also have an effect due to decrease of food supply. The suggestion

that the first born of the mother would be better nourished and

fewer wings would appear than in those born at the end of the

reproductive period is not supported, counter-evidence appearing in

the larger number of winged forms in the offspring of the older

parents. In experiments on winged mothers, starvation seemingly

had little effect. It thus appears that reduction of food supply is

the chief factor in bringing about the appearance of wings in the

progeny of wingless mothers. “The wing anlage appears to be

present in all the parthenogenetic females and depends directly

AMERICAN MICROSCOPICAL SOCIETY 279

upon the condition of the food supply of the mother for its stimu-

lation or suppression of development.”

Olfactory Organs of Lepidoptera.— McIndoo (717, Journ.

Morph., 29:33-54) made a morphological study of the olfactory

pores of forty-three species of Lepidoptera distributed among nine-

teen families in order to determine whether these organs are better

adapted structurally to receive olfactory stimuli than are the anten-

nal organs. It was found that the olfactory pores of Lepidoptera

occur on the legs, wings, and, in some species, on the mouth parts.

The disposition of these pores is similar to that of Hymenoptera and

Coleoptera. Structurally, they resemble those of other insects,

allowing for certain slight variation. Since in these olfactory pores

the ends of the sense fibers come into direct contact with the air,

they are better adapted structurally to receive olfactory stimuli than

are the antennal organs which are covered with hard chitin.

Paut S. WELCH.

Kansas State Agricultural College.

PHOTOMICROGRAPHS IN COLOR

Mees (American Photography 1917; and English Mechanic,

Sept. 14, 1917) communicates from the research laboratory of the

Eastman Kodak Co. the process of making lantern slides repre-

senting photomicrographs of stained sections. The print is made in

stained gelatin instead of the usual process.

The process of making such a print is as follows: Lantern

plates (Seed or Standard plates are satisfactory) are sensitised by

bathing for five minutes in a 2% per cent. solution of ammonium

bichromate containing 5 c.c. of strong ammonia to the litre, the

temperature of the bath being not above 65° F. The plates are

then rinsed for two or three seconds in clean water, drained, and

dried as uniformly as possible, being kept in the dark during drying.

The sensitised plates are then exposed through the glass under

the negative to the light of an arc lamp, the average exposure being

about three minutes at 18 inches distance. Printing cannot be

done by daylight, or sharp images will not be obtained. The exposed

plates are then developed by rocking in trays of water at about

280 NOTES, REVIEWS, ETC.

120° F. until all soluble gelatin is removed. Under-exposure is

indicated by the high-light detail washing away, and over-exposure

by the film being insoluble to too great a depth. The plates are

then rinsed in cold water, fixed in hypo, and washed free of the

hypo. They are then ready for staining.

The staining is done with a one per cent. solution of dye con-

taining one per cent. of acetic acid, the dye being selected to simu-

late most closely the original stain of the section, the time of dyeing

being chosen so that the necessary depth is obtained.

When sections stained with two different colours are being

photographed, negatives are made through suitable colour-filters,

and are then dyed in the two stains and placed face to face so that

a two-colour slide is obtained.

Suppose a section is stained red and green. Two negatives are

made on panchromatic plates—one with a red filter, which will cause

the green to appear as clear spaces in the negative and will not

record the red, and the other with a green filter, which will record

the red and not the green. The slides made as described from

these in bichromated gelatin are stained—that from the red negative

with the original green stain, and that from the green negative with

the original red stain.

The filters required can be chosen from the set of filters for

photomicrography prepared under the name of Wratten M filters.

The choice of the filter is decided by visual trial under the micro-

scope, the filters chosen being those which most nearly absorb one

colour and transmit the other. Thus, photographing a section

stained with Delafield’s hematoxylin and precipitated eosin, the A

filter (red) shows no trace of the eosin, and gives a good, strong

negative of the hematoxylin. The B and C filters are used together

for the other negative, giving a blue-green colour and recording

the eosin and hematoxylin both fully, and from these two nega-

tives positives are made and stained with a blue and a red dye.

A SHORT METHOD OF PREPARING HISTOLOGICAL MATERIAL

Dr. L. W. Strong, Pathologist, Woman’s Hospital, New York

City, published the following modification of current histologic

methods in the Journal of the American Medical Association for

AMERICAN MICROSCOPICAL SOCIETY 281

June 17, 1916. This method was devised to shorten the time for

making reports on histologic specimens, and I find that it is in every

respect equal to the regular routine procedure. The time is reduced

to three days together with a considerable saving in labor and re-

agents.

Fix: 10% liquor formaldehyde in 80% alcohol, over night.

95% alcohol, 8-10 hours.

Acetone, from one-half to two hours.

Chloroform-paraffin, over night in warm place.

Paraffin, four hours. 48° C., m. p., 2 hours; 52° C., m. p., 2 hours.

Embed.

Nm kW pw

H. R. Eqcieston.

Marietta College, Marietta, Oho.

ORGANIC EVOLUTION

This book is a comprehensive statement of the more important

facts and theories relating to organic evolution. It contains a per-

fectly stupendous amount of material, gathered and organized from

various sources, and is based on the author’s twenty-three years of

teaching. The excellence of the contribution is in this organization

rather than in originality of conception or statement. It is quite

safe to say that the result will prove a boon to the student and

teacher, as a carefully worked out compend.

This volume differs from the general run of such books pub-

lished in the last fifteen years, by its larger emphasis upon the geo-

logical illustrations and its fuller treatment of the palzeontological

data.

The book is divided into three parts. Part I includes six

chapters on the history of the idea of evolution; the organic king-

dom; classification; geographic, bathymetric and geological distri-

bution. Part IJ, under the general title “Mechanism of Evolution,”

treats in six chapters such subjects as natural, sexual and artificial

selection, variation and mutation, heredity, inheritance of acquired

characters, orthogenesis and kinetogenesis. Part III, of twenty-six

chapters, entitled ‘Evidences of Evolution,” is sub-divided into

three sections: ontogeny; morphology and adaptations; and Pale-

ontology.

282 NOTES, REVIEWS, ETC.

The evolution of the vertebrates is discussed at considerable

length, including chapters on the reptiles, the birds, the archaic

mammals, the modern mammals, the carnivores, the proboscideans,

the horses, the camels, and man.

It is well indexed and illustrated.

Organic Evolution, by Richard Swann Lull. Lllustrated by 253 figures; 729 pages.

The Macmillan Co., 1917.

ORIGIN AND EVOLUTION OF LIFE

In this book Osborn undertakes to deal with evolution in terms

of energy,—of action, reaction, and interaction,—rather than in

terms of mechanism. Life, whether simple or complex, is chiefly

concerned with capture of energy, storage of energy, and release

of energy. Mechanism has taken form about these physicochemical

processes. The physicochemical actions and interactions entering

into life-phenomena the author sums up as follows: (1) those

of the inorganic environment; (2) those of the developing organism ;

(3) those of the germ or heredity chromatin, and (4) the life en-

vironment.

Evolution is proceeding in each of these realms, and the sci-

entific and philosophic problem is to discover how these four evolu-

tions interact and are adjusted to each other. In particular it is

to be determined how the evolution of the germ plasm is influenced

by and adjusted to the changes in the other three.

The treatment of life origins revolves about the following

proposition: “Every physicochemical action and reaction concerned

in the transformation, conservation, and dissipation of energy pro-

duces also either as a direct result or as a by-product a physico-

chemical agent of interaction which permeates and affects the organ-

ism as a whole or affects some special part. Thru such interaction

the organism is made a unit, and acts as one, because the activities

of all the parts are correlated.” And thus as the evolution of action

and interaction, of receptors and effectors, has proceeded there has

been a corresponding evolution of interaction which is responsible

for the harmony within the organism. Heredity becomes, in this

conception, an outcome of the evolution of interaction.

AMERICAN MICROSCOPICAL SOCIETY 283

As instance and illustration of such regulating and balancing

of actions and reactions may be cited internal secretions, products

of life reactions which circulate in the system and secure coordina-

tion of activities at various points within it.

Clearly the present state of our knowledge will allow only the

most elementary elaboration of such a thesis. The author under-

takes specifically to suggest the manner in which the lower organ-

isms,—bacteria, alge, etc.,—capture electric energy, heat energy, the

energy of sunlight in the building up of the form of living substance.

In the second part of the book the evolution of the animal form

is traced in invertebrates and chordates as shown in the geological

record, in an effort to illustrate how adaptive form may arise in

connection with the essential actions, reactions, and interactions.

In this enterprise the author develops advantageously his well known

principle of adaptive radiation in the external body form as illus-

trated in vertebrate evolution.

All zoologists will be grateful to the writer for putting the

familiar material from a new point of view. It is a compelling

and stimulating statement. More than any recent writing on the

subject, this book synthesizes the evolutions of astronomy, geology,

physics, chemistry and biology.

The book is a beautiful and satisfying product of evolution, both

in respect to its energy and its mechanical form.

The Origin and Evolution of Life, by Henry Fairfield Osborn. Illustrated, 350

pages. Charles Scribner’s Sons, 1917. Price $3.00 net.

THE MICROSCOPE

This classic manual, produced by our long-time member and

past president, Professor S. H. Gage, is now in its twelfth edition.

This edition, which is largely revised, differs from previous ones

chiefly in the omission of matter pertaining to micro-chemistry and

metallography, and in the up-to-date treatment of those elaborations

of processes and apparatus that have taken place during the last

ten years. These include especially—the single objective binocu-

lar; the dark-field illuminator ; apparatus making possible the appli-

cation of powerful electric lights for demonstration and drawing ;

284 NOTES, REVIEWS, ETC.

improvements making for the development of photo-micrography ;

and the modification of artificial light to day-light perfection by

glass filters.

There are twelve chapters covering the usual range of the

microscopy. Each chapter is followed by a list of collateral ref-

erences, and a general bibliography concludes the book. Chapter

XII gives a brief history of lenses and other optical portions of the

microscope.

The user of Professor Gage’s book profits by the fact that

the writer is always alert to the practical bearing of any develop-

ment upon actual successful laboratory use rather than concerned

with the mere physical_or theoretical perfection of the device.

The Microscope, an Introduction to Microscopic Methods, and to Histology; by

Simon Henry Gage. Twelfth Edition; 472 pages, 252 figures. Comstock Publishing Co.,

Ithaca, N. Y., 1917. Price $3.00.

HPSS Pose Meiers NON Se REO Te eae

NECROLOGY

Information has come to the Secretary of the death of the fol-

lowing members :

Brown, Dr. Amos P., ’1l....... Germantown, Pa.

James Or. Wi Mi LZ ie ee ies Philadelphia, Pa.

COrentty AL Vi Oheiiaareevanusouen ',..Cleveland, O.

NSISHTING, (ite) Be LU ie seat Boston, Mass.

Ward, Dr. R. Halstead (Charter and Honorary

IEGINDEL) 0). Viner yiwraaner ep starale aunlaln th TroyyNay.

More extended notices will appear later.

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LIST OF MEMBERS

HONORARY MEMBERS

Crisp, FRANK, LLB, B.A. F.R.M.S.,

5 Landsdowne Road, Notting Hill, London, England

PFLAUM, MAGNUS Cai acls Soa be okies a eae alan paatnind ad as Mouttal Meadville, Pa.

LIFE MEMBERS

Brown, J. STANForD, Ph.B. A.M., P. O. Box 38, Far View, Black Hall, Conn.

CAPE SETH DUNRERICislc cviy dees veaas eens P. O. Box 2054, Philadelphia, Pa.

DUNCANSON, Pror. Henry B., A.M....... R. F. D. 3, Box 212, Seattle, Wash.

HULIOTT HPROP ARTHUR FL. scion oe adapts es 52 E. gist. St., New York City.

FR AMer PP TOH Nee ihe tia fale ods e'nie a Sesame Chicago Beach Hotel, Chicago, III.

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 ADMITTED SINCE THE LAST PUBLISHED LIST

Arnold, Wm. T. Page, I. H.

Busch, K. G. A. Pohl, John C., Jr.

Bunker, Geo. C. Roe, G. C.

Burke, G. E. Stephens, E.

Chambers, W. E. Stewart, T. S.

Frisch, John A. Taggart, R. B.

Hall, F. Gregory Warner, E. A.

Hitchens, Alfred B. Van Cott, H. A.

ACKERT, JAMES EDWARD, 'II............ Kas. State Ag. Col., Manhattan, Kas

ALLEN, HARRISON SANBORN, M.A., ’15, 442 Farmington Ave., Waterbury, Conn.

ALLEN, Wo. Ray, M.A, ’15...... 212 So. Washington St., Bloomington, Ind.

Atien| Wywrrep: Fi AcMs Vos lie vias ors siniec e's a e's High School, Fresno, Cal.

ADDERSOM p UM BEATING IO ira Rea ali Gly ea uta Station A, Lincoln, Nebr.

AnmpgaAs CoB AS ig er On cue awn ate ete 540 S. Main St., Manchester, Ill.

AUWOLD RAN 1A. iit cetee gaure itu wis ech 408 House Building, Pittsburg, Pa.

ARNOLD, W BEY Tio FTF wits Gs ela lek Ue Fa aieiats 702 Penn Ave., West Reading, Pa.

ATHERTON, Pror. L. G., A.B., M.S., ’12..State Normal School, Madison, S. D.

PUPMOOD, FA. Via 7O wanie un vie wien cokes Kore eh 16 Seneca Parkway, Rochester, N. Y.

Batpwin, Herpert’ Boge 35 Vis va etek cea 927 Broad Street, Newark, N. J.

BARKER, FRANKLIN D., Ph.D., ’03..... University of Nebraska, Lincoln, Neb

Sager WBS) Manas. Dumwwwrats sees sana: Clemson College, S. C.

acai. G MED. crs oe ware a eae 3515 Prytania Street, New Orleans, La.

Bauscu, EpwArb, ’78........66 steht sie: 179 N. St. Paul St., Rochester, N. Y.

288 LIST OF MEMBERS

BAUSCRy \Wittram, 8S, 1 0 We pil ik eg) E St. Paul St., Rochester, N. Y.

SRE ACUI UIMAS ree kuna kari 2811 Benvenue Ave., Berkeley, Cal.

Beck, WiLLiAM \A.,-M:Sc., 716. 8) St. Mary College, Dayton, Ohio

Bett, Ausert T., B.S., PAL IMS NE eae La. State Univ., Baton Rouge, La.

DEN NEHOPE, J. 1): MASH tay Lu i Alfred College, Alfred, N. Y.

BENNETT, Henry C., ’93....Hotel Longacre, 157 W. 47th St., New York City

BESTS, JOR NB ea icn Mona Cou ay AW Lat 111 Market St., Camden, N. J.

Binrorp, Raymonp, Ph.D., ’15............. 226 College Ave., Richmond, Ind.

Birce, Pror. E. A. ScD., LL.D., DOK a Cas eR 744 Langdon St., Madison, Wis.

BLACK, Jy EtG Woe OR IN OT LWA Nua) OL: 530 Wilson Bldg., Dallas, Texas

Bree ARMM Pag PR ity: Ohio State University, Columbus, Ohio

Bootu, Mary A., F.R.M.S., BORE By Bae Ay ee Eb ul ee a a

AOS We 4 'shba Wide avid Bata OM MMCRREGMN, | Ca 60 Dartmouth St., Springfield, Mass.

Bovey Ce Suit: Mi eae 6140 Columbia Ave., Philadelphia, Pa.

Brope, Howarp S., Ph.D., ’13....... 433 E. Alder Street, Walla Walla, Wash.

Brookover, Cuas., A.B., M.S., WS ag Univ. of Louisville, Louisville, Ky.

BRowny FR, ALB! feat BA ba William Nast College, Kiukiang, China

BROWNING, S1ipNEY Howanrp, ’I1. -Royal London Ophthalmic Hospital, London

Brunn, Cuarues A., LL.B., ’16.............. 14 E. 56th St., Kansas City, Mo.

Bryant, Pror. Eart R., A.M. ’Io...... Muskingum College, New Concord, O.

BuckInNGHAM, Epwin W., J8.,................ Va. Med. Col., Richmond, Va. ”

Buty, JAMES Epcar, Esg., ’92................. 141 Broadway, New York City

Butuitt, Pror. J. B., M.A,, SAB Mutha 1 MBN es (AR Mr Aly Ly fy Chapel Hill, N. C.

TOO ASE GS te CUE nA nH ae Gatun, Canal Zone

BUSCA) HOARL GUA A BL ar i uiganes oe ie hih Bixley Station, Columbus, O.

131504 ion Cau ONES G ORMUNMLN, ARMA He U. S. Naval Hospital, Las Animas, Colo.

BUS WELL ALM MAS 6h. Tae at Columbia Univ., New York City

CABALLERO, Peon.) GUSTAV (AL M16). 00 Ul. 4! sous dn uty ick kta at Or

PUPS Pa LEE MARL AL Ua ARN doulas pu ALLA a Dh be Fordham Univ., New York City

CAMPBELL, JoHN PENDLETON, Ph.D, ,13’... University of Georgia, Athens, Ga.

Gaaeson, (CoM HWA Bulsa lieu inthe foe Doane College, Crete, Nebr.

Caarrm, Prov, Ciarnes reiytiey yt eue le Parsons College, Fairfield, Ia.

Carter, JoHN E., ’86.......... 5356 Knox St., Germantown, Philadelphia, Pa.

Cuampers, W. E,, ’17.............. Dept. of Agriculture, Washington, D. C.

CHESTER, WAYLAND Morcan, M.A., ’15..Colgate University, Hamilton, N. Y.

Cuickerinc, A, M., A.M., '16............... 1141 Harrison Ave., Beloit, Wis.

CiarK, Grorce Epw., M.D., MOY Grae edd ace te hee Genessee St., Skaneateles, N. Y.

Crags, Howars We A:Mi trac. vhU Cn Nima ie Fairport, Iowa

CLEMENTS Magy! Fo Ph vitamin lid ahi a0 WL eae Tuscon, Ariz.

Com, Bu ALP h Oe a au LON cone Falls Church, Va.

CocHILL, Pror. Georce E., ed oh GI a R. F. D. 9, Lawrence, Kas.

Cotton, Haroxp S., Ph.D., ’11... -Zoological Lab., Univ: of Pa., Philadelphia

Con, (ALB irs WP NV MU Editorial Staff, “Lumberman,” Chicago, IIl.

AMERICAN MICROSCOPICAL SOCIETY 289

Concer ALLEN Ga MUA TS eed lsisisle'ele's P. O. Box 663, East Lansing, Mich

CCONLON (PASS IE TE De a ance arses 717 Hyde St., San Francisco, Cal.

Cooper, Artuur R., A.M., ’16....College of Medicine, Univ. Ill. Chicago, Ill.

CorNELL Univ. Lrprary (Pror. S. H. GAGE)..........-e ee eeee Ithacay: NivY;

CORT WV TeGe ne she ts o's k o2 Dept. Zool., U. of Cal., Berkeley, Cal.

Corr GRORGE BT oi ace tile ene agen eke 1001 Main St., Buffalo, N. Y.

Govier): Gromae Wie Piri y sess ea k ba Sue e ile waely College View, Nebr. ,

CROZIER, WILLIAM JOHN, PH.D., 715......cccceccccc cece esccerescsces

SPE Teh cea See ies Bermuda Biological Station, Agar’s Island, Bermuda

Darpaker, LEASuRE Kine, Ph.D., M.D., ’I1...... ce cece ee eee eee ee eees

Jane We aie Pe Qe aide id AMP a! 7025 Hamilton Ave., Homewood Sta.

DAVIS RON Eloy Rady) 02.) ok oe University of Florida, Gainesville, Fla.

Deere, Emit Oxar, A.M., S.M., ’13....Bethany College, Lindsborg, Kans.

DeWitt, Cuarres H., M.S., 711......0.00-- 355 College Ave., Valparaiso, Ind.

Disprow, WittiaM S., M.D., Ph.G., ’ol....... 151 Orchard St., Newark, N. J.

Teck, GARBOLE. Wiad. 6h Ni 0s Se gae se se Mo. Bot. Garden, St. Louis, Mo.

IDOLBEY, -UWARD Pag 060 fi SURE 3613 Woodland Ave., Philadelphia, Pa.

DOLE Fi WILBURS Dt Oy 13h. Vedi t ee desead ieee dices eal dads Fairfield, Iowa

DousLeDAY, ARTHUR W., M.D., ’16...... 220 Marlborough St., Boston, Mass.

DrescH_Er, W. E., ’87........ Care Bausch & Lomb Opt. Co., Rochester, N. Y.

Dusss, Lewis ALBERT) "16.0026 oc esce eens Kas. S. Ag. Coll., Manhattan, Kas.

TUpcKoN; WINFIELD! BIS) PPE eee on ae aie 932 E. 55th St., Chicago, Ill.

Duncan, Pror. F. N., Ph.D., ’16.......... So. Methodist Univ., Dallas, Tex.

Epmonpson, Cuarues H., Ph.D., ’15........--+- 1360 Alder St., Eugene, Ore.

POD UMICTONG Wh ER ern ide Cine inate aden ay wt elustoaas State College, Pa.

Eppy, (SAMUEL AL, 715. cece rs ote toe bet «sein ge oe omen deka ee Tower Hill, Ill.

Eecoteston, Hi RMA vis. ceases cee Marietta College, Marietta, Ohio

E1cENMANN, Pror. C. H., ’95........66> 630 Atwater Ave., Bloomington, Ind.

Exiiotr, FRANK R., M.A,, "15... .ccccccecescssecececceces Wilmington, Ohio

Exzis, Pror. M. M., Ph.D., ’12.......cecesseeees 1109 13th St., Boulder, Colo.

Errop, Pror. Morton J., M.A., M.S., 08........2eee eee eeecee rere eeeees

a ary, PIAA e cnet Deena erg SMe ET has University of Montana, Missoula, Mont.

BOWELL As 1a. SOcbs i eaacehabsveatenea te 210 The Normandie, Seattle, Wash.

EssEensBerc, Mrs. CuRIstINnE, M.S., ’16........ Scripps Institute, La Jolla, Cal.

ESTERLY, CALVIN O,, 'I5......ccccccccsoocees Scripps Institute, La Jolla, Cal.

Eyre, Joun W. H., M.D., M.S., F.R.MLS., '90..... 0 eee ce cere ee eee eens

Tak Haleue hs oot ee slo ee eee aa Se Guy’s Hospital, London, E. C., England

Fasrow, Prov, W.\G., 11 ei case eeetaaees 24 Quincy St., Cambridge, Mass.

Fatric, Pror. P. W., B.S., M.S., 12... cece see e eee c cece eer eeeeereneees

Ba ah ac be ROA aed eae State Normal School, Valley City, N. Dak.

Fetz; Gro. E., M.D., P.RiM.S, (18.0008. 309 Porter Ave., Buffalo, N. Y.

Fettows, Cuas. S., F.R.M.S., ’83...107 Cham. of Comm., Minneapolis, Minn.

Fercuson, Marcaret C., Ph.D., ’11..... ',..Botanical Dept., Wellesley, Mass

FERNANDEZ, Fr. MANUEL, B.S., ’16.San Juan de Latran College, Manila, P. I.

290 LIST OF MEMBERS

PINDLAY, (MERLIN CG. AyMy, Urs oie, canals Park College, Parkville, Mo.

PSC MERON IAE I OBL s'claty's e Wicg hil a wie ele eae ie is Wi Box 1608, Milwaukee, Wis.

HIT7Z-RANDOLPH, RAYMOND (Ba RR UMS) ir ce Ce Na ae

Pa Pa eee Wid aig Wide m NRE State Laboratory of Hygiene, Trenton, N. J..

Fiint, JAMES M., M.D., ’or............. Stoneleigh Court, Washington, D. C

BOOTED Se EDN Ora Re a Bek Griese 202 S. Thirty-first Ave., Omaha, Neb.

POSTERS VVILETAM CNS OIG On mn til 707 Coleman St., Easton, Pa.

PURNISS, Ee WieM Da PR Di toe OL ane 52 Vernon St., Hartford, Conn.

RTABBIEE MO PTO WNL Oy ty Mm tO 2659 California St., San Francisco, Cal.

Gace; Prov: Staton Hy BLS Bae yea 4 South Ave., Ithaca, N. Y.

GaALLOwsy, (Pron Ty Wes AIMS PRD orien Ca eek a No Nae Beloit, Wis.

GARRETSON) ISUGENE, [120400 ye), 428 Fargo Ave., Buffalo, N. Y.

GESNER, BROWER CLAIR, ’II........ 110 Steadman St., Moncton, N. B., Canada

GOLDSMITH CaO AU tah tas tiaturs S. La. Indus. Inst., Lafayette, La.

GOWENAERANCTS (HL Git ay ali ies Cn ew R. D. 1. Box 15, Exeter, N. H.

GRAHAM, CHARLES W., M.E., ’II..........%. 447 W. 14th St., New York City

GRAHAM) JOHN YOUNG, (PhDiiita iii seh ceed eeeus University, Alabama

LOY he gh dh OAC 6 7 ZUMA RAL ap RERUN panne Wat a 3535 Telegraph Ave., Oakland, Cal.

GRAY WILETAR \CACCEN DA EU eas Nis aN ie ae Lock Box 233, Tama, Iowa

GRIFFIN, LAWRENCE E., ’13.......... University of Pittsburg, Pittsburg, Pa.

GUTBERLET, JOHN E., Ph.D., ’11............ Carroll College, Waukesha, Wis.

Guyer, Micuaet F., Ph.D., ’11....... University of Wisconsin, Madison, Wis.

TEAGECSTEUN ROBERT 1 1G 5 RGU ORLA Mra oe Minneola, Nassau Co., N. Y.

PAGIReO Be MD rota OOP e nia Hagler Building, Springfield, III.

TTAGUE, PLOUENCE LAME. Whcy an ieena lope Nat. Hist. Bldg., Urbana, II.

TEAR AD Re COR Nu Sy Re OO Ay Ea oR Aa Milton College, Milton, Wis.

Hance, Rosert T., B.A., ’13........ Zool. Lab., U. of Pa., Philadelphia, Pa.

BEAN EIN BOM OCB CRTs MOOI T eCGOT TY 2 MEW atm arta Nyy oor Charleston, Ill.

HANNAH, MarGareT L., A.M., ’16............00005 Station A., Lincoln, Nebr.

ELANSEN DAMES! (RR io aan pon St. Johns Univ. Collegeville, Minn.

FEARDY PEUCESI ET Sine Ga Aee ae ek ee cae eed 1860 12th Ave., Moline, III.

HarMAN, Mary T., ’13....... Kansas State Agr. College, Manhattan, Kansas

Hayorn) ogace Bawiny Jeo ta ad ics ar ae We College Station, Texas

Harpies PR Depeche logan Wash. State College, Pullman, Wash.

HEIMBuRGER, Harry V., A.B., ’14........ 1843 Feronia Ave., St. Paul, Minn.

HENDERSON; WILLIAM, TED UO ei deena Millikin Univ., Decatur, Ill.

HertzoGc, MAxMILIAN, M.D., ’or.......... 3152 Cambridge Ave., Chicago, III.

FUE TON WAL ETA MA PID rr ae is fete a ie el aun ta Claremont, Cal.

FISSSONG) ROY ELD! BS 2 16 ee OO TA al aa Madison, So. Dak.

HircHins,, Aurrep B:,\ PhD.) '17.05..5 0 00s 2 Dwight Block, Binghamton, N. Y.

Hyorrm) Lboupwie iC. hi rend Meadowdale, Snohomish County, Washington

Hoty Cross CoLLecre, PRoFESSOR OF BIOLOGY................ Worcester, Mass.

Hosmins, (Wiatiloo ena lan Van teh LN ca 49 6th St., LaGrange, Til.

Howarp, Rosert Nessit, ’12..Ookiep, Namaqualand, Cape Province, S. Africa

AMERICAN MICROSCOPICAL SOCIETY 291

How anp, Henry R., A.M., ’08.............- 217 Summer St., Buffalo, N. Y.

FINGHESHIOALEN Ey LS Oiiod cle etd silt ales pita Grinnell College, Grinnell, lowa

TyPS RRR tC COS elation cig:d also devernere ey ye 1201 Race St., Philadelphia, Pa.

JERFS) PE ROR en LIMO pew sind Weel de altars Univ. of Okla., Norman, Okla.

JENNER Ee ere lace kines el suns aeaecioweuss Science Hall, Indianola, la.

JENNINGS, HENRY RALPH I5 fo ide 8 velreeieiets Freemont H. S., Oakland, Cal.

NOE NSONG Dy Nie Lec ere e Wee. Woh wisielelel diengter bein Joplin, Mo., R. F. D. 4-147

JOHNSON, Crake bey Dan EIR iG. ee ee 200 W. 72nd St., New York City

JOHNSON; PRANK Sci MD 03s ly Seis oe 2521 Prairie Ave., Chicago, IIl.

JORDAN, ROR TE kay (AS vee dy 6 sisal’ a's University Place, Charlottesville, Va.

JUDAY) GHARCEY) OO ey veces ddeds Biology Bldg., U. of W., Madison, Wis.

FRIRIGAM Td AR a ROTO NEY clits oot oe etn atace St. Procopius College, Lisle, Ill.

BERD Tet eke gOs ce cdieslevele shane 202 Manchester St., Battle Creek, Mich.

I PRNALL MORRIS PANE 88 os FRU aes Oe a PE TOS OF OU pe Bismark, No. Dak.

Kincarp, Trevor, A.M., ’12........ University of Washington, Seattle, Wash.

pC EST ORI Bsp OF AUN S UNSERE pa 9) (0h Mia a ORR SU RIT ls NRE a ALG TAR A Centerville, Iowa

PENG VV ILLIARINEW DOE Cele bss bier whe ens P. O. Box 261, New Orleans, La.

Kirscu, Pror. ALEXANDER M., M.G., ’16.......... Notre Dame (Univ.), Ind.

PNIGH TINE Ue EE AUT Die tag se bin Chai ee ek he ei ke 1015 Blondeau St., Keokuk, Ia.

Koroip, Cartes A., Ph.D., ’99....... University of California, Berkeley, Cal.

Korg Aa eM DeVOn or Win ditveena maleate ailaldts 32 S. Fourth St., Easton, Pa.

KRrecKER, FrepERIC H., Ph.D., ’15..... Ohio State University, Columbus, Ohio

Franck, LOUIS) Misi hain vesueyneiaen nea 520 Elm Street, San Jose, Calif.

LACY A PRANKUW 3 Sse eee kG U. S. Naval Hospital, Las Animas, Colorado

LAMBERT, GilALy IS 0 78 Say ee PARTE Sl oe based eewawels Bae ale iblele ofala aa wae

Eta URE Bank of New South Wales, Warwick, Queensland, Australia

LAND, WILLIAM JESSE GOAD, Ph.D., ’15...--ceeccecesececesccrrerecces

SLES AN TESTO ROS AM a OS By The University of Chicago, Chicago, IIl.

TANNER CIA VAN Vaipicete Bae Cok did aketinhan 6 waaay Univ. of Okla., Norman, Okla.

GAN TZ CYRUS OW PAM DET aN abate diate odoin ints ata’s University, Reno, Nev.

LaRur, Georce R., Ph.D., ’11....University of Michigan, Ann Arbor, Mich.

Laruam, Miss V. A., M.D., D.D.S., F.R.M.LS., ’88.......ccseseeeececocs

nh AN Ie tyA onal arg We) DON AN AS SN 1644 Morse Ave., Rogers Park, Chicago, IIl.

Latimer, Homer B., M.A., ’11......... Le ei 1226 So. 26th St., Lincoln, Nebr.

LEHENBAUER, Pup, A.M., ’I1...... Floriculture Bldg., U. of I., Urbana, III.

Lewis, Mrs. KATHERINE B., ’89............. Bellwood Farms, Geneva, N. Y.

ERWISS Lk RST ica a Chats ea eaten Okla. Ag. Exp. Sta., Stillwater, Okla.

Lirrerer, W:., A.M., MiD., 700.000) cece eects ces ececesessws Nashville, Tenn.

Lome ADOLPH, (Of: cedpaccesss ves 289 Westminster Road, Rochester, N. Y.

LoncFELLow, Rosert CaPLes, MSM De eros eens 1611 22nd St., Toledo, O.

Paver, HUGH Bis kien sice ce sick ak Gets a eee 2120 High St. Denver, Colo.

Lyon, Howarp N., M.D., ’84..........0+- 828 N. Wheaton Ave., Wheaton, IIl.

MacGitiivray, ALEXANDER D., ’12....603 W. Michigan Avenue, Urbana, IIl.

Mack, MarGARET EvizABETH, A.M., 713.....--. sere eee eee eee Dayton, Nevada

292 LIST OF MEMBERS

INEAA THORS aoe! AMS, ISS ih wae e Gay's Medical College, Us of I., Chicago, III.

Marr, Georce Henry, M.E., ’rt............. 94 Silver St., Waterville, Maine

MarsHALL, Couns, M.D., ’96........... 2507 Penn. Ave., Washington, D. C.

MARSHALL, Rute; Ph.Ds 07) 1.65 cies ba pales Lane Technical H. S., Chicago, Ill.

RRBRSHALL We S. PT e 1S inca uci Cousens 139 E. Gilman St., Madison, Wis.

MartTLANpD, Harrison S., A.B. M.D., ’14...... 1138 Broad St., Newark, N. J.

BLABSEY; PROB tdi tet lade « cade chev ou tae Clemson College, S. C.

PMATHER, FMD GPh Dy 262 os enon 228 Gratiot Ave., Mt. Clemens, Mich.

MAY, bibarey Gustay, Bis 775s ras ce cata iie pak dhs wl Rh ak Bee

Ae RTOs SHG eS th We IN Bur. Animal Industry, Zool. Div., Washington, D. C.

MAYHEW) Rov LotR S. STS eAILe uantat eee 1156 W. Decatur St., Decatur, Ill.

MAYWALD, FREDERICK J., ’02........ 1028 Seventy-second St., Brooklyn. N. Y.

McCatrra, Anpest, Ph.D.) "Bort. oi). sce ses 2316 Calumet Ave., Chicago, III.

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INDEX

Achert, J. E. and A. A. Grant. Cestode

from Young Cat, 93

Acid Colloidal Dyes in Blood and Tissue

Cells, 110

Adipose Cells, Amitosis in, 274

Aleurodes Parthenogenesis in, 273

Algz, Synopsis of the Blue-Green, 179

Amitosis in Adipose Cells, 274

Ameeba, Culture of, 167

Amceba, Growth of on a Solid Medium,

Animal Micrology (Review) M. F.

Guyer, 48

Another Cestode, from the Young Cat,

Anthothrips, Sex in, 274

Apparatus for Gradual Dehydration, 27

Bacteriology in Plant Pathology, 5

Barker, Franklin D., A Monostome

Lung Fluke from the Painted Terra-

pin C. Marginata, Agassiz, 55

Bath, An Individual Paraffin, 32

Biology of Fireflies, 100

Birds, Laying Cycles in, 107

Birds, The Fundus Oculi of, 172

Bishop, G. H., Apparatus for Gradual

Dehydration, 32

Blood Cells in Vitro, Insect, 98

Blood and Tissue Cells, Acid Colloidal

Dyes in, 110

Bone Sections, Pyridine Silver for, 272

Blue-Green Algz, Synopsis of the, 179

Brown, Dr. Amos P., 285

Castration of the Vertebrate Embryo,

Early, 106

Cervical Region, Nature of the, 276

Cestode from the Young Cat, 93

Chamot, E. M., Chemical Microscopy,

139

Chart Making, 269

Chemical Microscopy, 139

Chick Embryos for Demonstration,

Preparation of, 47

Chironomus, Hemoglovin in, 97

Chrisemys Marginata, Agassiz, A Mono-

stome Lung Fluke from the Painted

Terrapin, 55

Chromosomes, Fixation of Mammalian,

108

Chromosomes, The Segregation and Re-

combination of, 106

Chroococcacez, 185

Chrystallizable Chemical Salts, Photo-

micrographs of, 41

Ciliated Epithelium for

Study, 47

Classification of Diptera, 99

Clearing and Mounting Hydra, 46

Clearing Method, Potash, 46

Clearing Method, Substitute for Spalte-

holtz, 46

Cold on Malaria Parasites, Effect of, 98

Color, Photomicrographs in, 279

Color Markings, Development of, 98

Colloidal Dyes in Blood and_ Tissue.

Cells, 110

Comparison Between Nuclei of Nerve

Cells, 157

Conjugation, Are Encystment and Con-

jugation Necessary, 170

Contractile Vacuoles in Paramecium,

170

Cooperative Technique, 108

Culture of Ameeba, 167

Culture of Euglena, 168

Culture of Pristina, 169

Histological

300

Culture of Tetrastemma, 168

Cytological Changes Accompanying Des-

sication in Rotifers, 171

Dean, Carlton F., An Easily Adjusted

Imbedding Box, 38

Dermested Larve, Inanition of, 277

Dessication in Rotifers, Cytological

Changes Accompanying, 171

Development of Color Markings, 98

Development of Genital Glands, 100

Development of Malpighian Tubules,

277

Diptera, The Head and Mouth, Parts

of, 105

Doubleday, Arthur W., Photomicro-

graphs of Crystallizable Chemical

Salts, 41

Drosophila, Reactions of, 97

Early Castration of the Vertebrate Em-

bryo, 106

Eggs, A Method of Cutting Planorbis,

Electrifying the Microtome, 267

Embryonic Development, Protoplasmic

Continuity in Early, 103

Embryos of Chick for Demonstration,

Embryos, Early Castration of the Verte-

brate, 106

Enchytreide (Oligocheta) of

Woods’ Hole Region, 109

Enchytreide (Oligocheta), From the

Rocky Mountain Region, 69

Enchytreus Albidus, 120

Encystment, Are Conjugation and En-

cystment Necessary, 170

Entomological Notes, 273

Epithelium, Ciliated for Histological

Study, 47

Eugenics, Genetics and, 110

Euglena, Culture of, 168

the

INDEX

Euglena, Quince Jelly as a Culture,

Medium for, 109

Evolution of Life, Origin and ( ARE,

282

Factors Controlling the Rate of Regen-

eration, 104

Factors Influencing the Sporangial

Characters of Mycetozoa, 103

Fireflies, Biology of, 100

Fish Without Ice, Preserving, 170

Fixation of Mammalian Chromosomes,

108

Friderica Ratzelli, 130

Fundus Oculi of Birds, 172

Further Notes on the Rearing of Vol-

vox, 271

Genital Glands, Division of, 100

Genetics and Eugenics, 110

Gland and Pigmentation, The Pineal,

107

Gradual Dehydration, Apparatus for, 27

Grant, A. A. and J. E. Ackert, Cestode

from Young Cat, 93

Growth of Ameeba on a Solid Medium,

Hemoglobin in Chironomus, 97

Head and Mouth Parts of Diptera, 105

Head of Insects, Evidence as to the

Number of Segments in, 48

Henderson, Robert W., An Individual

Paraffin Bath, 36

A Microscopic Method of Reconstruc-

tion, 38

Henlea Urbanensis, Welch, 75

Hibernation, 276

Hilton, Wm. A., Some Comparisons

Between Nuclei of Nerve Cells, 157

Histological Material, A Short Method

of Preparing, 280

AMERICAN MICROSCOPICAL SOCIETY

Histological Technic, 270

Hitchins, Alfred B., A Device to Coun-

teract the Effect of Vibration on

Photomicrographs, 40

Hydra, Clearing and Mounting, 46

Imbedding Box, An Easily Adjusted, 38

Inanition of Dermested Larve, 277

Injections of Semi-circular Canals, 46

Insect Blood Cells in Vitro, 98

Laying Cycles in Birds, 107

Lepidoptera, Olfactory Organs of, 279

List of Members, 287

Lumbricillus Lineatus (Mull), 123

Lung Fluke from the Painted Terrapin

Chrysemys Marginata Agassiz, 55

Microscopic Method of Reconstruction,

36°

Malaria Parasites, Effect of Cold on, 98

Malpighian Tubules, Development of,

277

Mammalian Chromosomes, Fixation of,

108

Mesenchytreus Altus, 67

Methods Reported from the Zoological

Laboratory, University of Wisconsin,

Methods, Technical, 267

Microscopy, Chemical, 139

Microscopic Organisms for the Zoolog-

ical Laboratory, Notes on the Cultur-

ing of, 163

Microscope, The, Gage (Review), 283

Microtome, Electrifying the, 267

Monostome Lung Fluke from the

Painted Terrapin, Chrysemys Margin-

ata, Agassiz, 55

Mounting and Clearing Hydra, 46

Mounting Medium, A New, 42

301

Mouth in Cases of Pyorrhea, Parasites

in the, 101

Mouth Parts of Diptera, The Head and,

105

Museum Methods, 13

Mycetozoa, Factors Influencing the

Sporangial Characters of, 103

Myxophycez, Synopsis of, 179

Nature of the Cervical Region, 376

Necrology, 285

Nelson, T. C., Methods from Tooleeeal

Laboratory, University of Wisconsin,

Nerve Cells, Nuclear Size in, 99

Nerve Cells, Some Comparison Be-

tween Nuclei of, 157

Neurology, An Introduction to (Re-

view), Judson C. Herrick, 49

Norton, C. E. A New Mounting Medi-

um, 42

Nostocacez, 187

Notes on the Culturing of Microscopic

Organisms for the Zoological Labor-

atory, 163

Nuclei of Nerve Cells, Some Compari-_

sons Between, 157

Nucleoli of Silk-gland Cells, 275

Nutting, C. C., Museum Methods, 13

Olfactory Organs of Lepidoptera, 279

Oligocheta, Enchytreide (——), from

the Rocky Mountain Region, 67

Oligocheta, Enchytreide (——), from

the Woods’ Hole Region, 119

Orcutt, A. W., 283

Organic Evolution (Review), Lull, 281

Origin and Evolution of Life (Review),

Osborn, 282

Origin of Photogenic Organs, 275

Oscillatoriacez, 186

302

Inf

Paraffin Bath, An Individual, 32

Paramecium, Effect of Thyroid on, 169

Paramecium, Extra Contractile Vacuoles

in, 170

Parasites, Effects of Cold in Malaria, 98

Parasites in the Mouth in Pyorrhea, 101

Parsons, Suzanne, A Monostome Lung

Fluke from the Painted Terrapin,

Chrysemys Marginata, 55

Parthenogenesis in Aleurodes, 273

Photogenic Organs, Origin of, 275

Photomicrographs in Color, 279

Photomicrographs of Crystallizable

Chemical Salts, 41

Photo-micrography, A Device to Coun-

teract the Effect of Vibration in, 40

Pigmentation, The Pineal Gland and,

107

Pineal Gland and Pigmentation, 107

Place, J. A., The Morphology, Struc-

ture and Division of MHydractinia

Polyclina, 83

Planorbis Egg, A Method for Cutting,

Plant Diseases of Economic Import,

Table showing progress of knowledge

concerning American, 11

Plant Pathology, Bacteriology in, 5

Potash Clearing Method, 46

Preserving Fish Without Ice, 170

Pristina Culture of, 169

Protoplasmic Continuity in Early Em-

bryonic Division, 103

Pyridine Silver for Bone Sections, 272

Pyorrhea, Parasites in the Mouth in, 101

Quince Jelly as a Culture Medium for

Euglena, 109

Reactions of Drosophila, 97’

Reconstruction, A Microscopic Method

of, 36

INDEX

Regeneration, Factors Controlling the

Rate of, 104

Report for Spencer-Tolles Fund 1916,

112

Richards, A., A Method for Cutting

Planorbis Eggs, 44

Rivulariacee, 188

Roberts, E. W., Evidence as to the

Number of Segments in the Head of

Insects, 48

Rotifers, Cytological Changes Accom-

panying Dessication in, 171

Scytonemacee, 189

Segments in the Head of Insects, 48

Segregation and Recombination of

Chromosomes, 106

Semi-circular Canals, Injection of, 46

Sex in Anthothrips, 274

Short Method of Preparing Histological

Material, 280

Silk Gland Cells, Nucleoli in, 275

Spalteholz Clearing Method, Substitute

for, 46

Spencer-Tolles Fund, Report for 1916,

112

Sporangial Characters of Mycetozoa,

Factors Influencing the, 103

Starvation and Wing Development, 278

Stevens, F. L., Bacteriology in Plant

Pathology, 5

Stigonemacezx, 187

Synopsis of the Blue-Green Algz, 179

Technical Methods, 267

Technique, Cooperative, 108

Teleosts, Vital Dyes in, 109

Tetrastemma, Culture of, 168

Thyroid on Paramecium, Effects of, 169

Tilden, Josephine E., Synopsis of the

Blue-Green Algzee—Myxophycex, 179

Tubifex, 101

| AMERICAN MICROSCOPICAL SOCIETY 303

V Welch, M. W., Growth of Amceba on a

Vibration, A Device to counteract the Solid Medium for Class Use, 21

effect of, in Photo-micrography, 40 Welch, Paul S., Enchytreidze from the

Vital Dyes in Teleosts, 109 Rocky Mountain Region, 67; Enchy-

Volvox, Further Notes on the Rearing treide from the Woods’ Hole Region,

of, 271 119; Entomological Notes, 97; Notes

W on Oligocheta, 100; Wing Develop-

Ward, Dr. R. Halstead, 285 ment, Starvation and, 278

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