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VOLUME XXVII

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EDITED BY THE SECRETARY

Twenty - Eighth Annual Meeting

HELD AT

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VOLUME XXxVII

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OFFICERS FOR 1905-1906

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Perens A, M. HOLMES 2... 0... ee ecb ee ccatccs dd: Denver, Col.

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Past Presidents still retaining membership in the Society

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at Indianapolis, Ind., 1878, and at Buffalo, N. Y. 1870.

J. D. Hyatt, of New Rochelle, N. Y.,

Apert McCatia, Ph.D., of Chicago, IIl.,

pe our.) PhyD.; of Urbana, IIl.,

at Chautauqua, N. Y., 1886, and at Buffalo, N. Y., 1904.

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at Detroit, Mich., 1890.

at Washington, D. C., 1801.

at Columbus, O., 188t.

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_ Frank L, James, Ph.D., of St. Louis, Mo.,

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

at Rochester, N. Y., 1892.

at Ithaca, N. Y., 1895.

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

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W. C. Krauss, M.D., of Buffalo, N. Y.,

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The Society does not hold itself responsible for the opinions expressed by

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TABLE OF CONTENTS

FOR VOLUME XXVII

The Annual Address of the President, The Relation of Animals to Dis-

ease, by Henry B, Ward's... 2.0.6.0) ss ve ede eeaeanaeaei ser eras ne 5

A Contribution to the Microscopic Study of Pen and Ink Lines, by Mar-

shall D. Ewell tic cc 2s cs» babes we cece ole « X= seek een reer ee 21

On the Occurrence of Trypanosomes in the Blood of Rana clamata, by

James H. Stebbins, Jr., with Plate I .......seseeeercewscewseceseee 25

Tapeworm Cysts (Dithyridium cynocephali n. sp.) in the Muscles of a

Marsupial Wolf (Thylacinus cynocephalus), by B. H. Ransom, with

LOUPE a hiey neers seseepennbawaavaeaanvirnce os 96 sahswh SPI meg sdoce Oe

Probstmayria vivipara (Probstmayr, 1865) Ransom, 1906, a Nematode

of Horses heretofore unreported from the United States, by B. H.

Ransom, with Plate II ......-.cesceseeceesscececeesecesseccccsess 33

Porocephalus constrictus in a native Filipino, by Maximilian Herzog and

Chas, Be Hare oi cb ick eee de ode teeth tens ce can sneneen tame eunamne 4I

The Structure and Classification of the Siphonales, by Charles E. Bessey,

with ‘Plate LIL o. is ci bewccesWeusecheck «un pacensesiunents ets SseatmnimeE 47

The Phylogeny of Bacteria, by F. D. Heald, with Plate IV.........-.+-- 63

A Biological Study of the Lakes of the Pike’s Peak Region—Preliminary

Report, by H. L. Shantz, with Plates V, VI, and VII.........--+-++> 75

Variations in the Vitellaria and Vitelline Ducts of three Distomes of the

Genus Opisthorchis, by F. D. Barker, with Plates VIII and IX...... 99

Some Studies on Trypanosoma lewisi, by Leroy D. Swingle, with

Plate Xciscaccccdccsccesdenequat fp pisces we kn st pee) t6Ré ae nse mer mEE III

New Forms of Volvox, by J. H. Powers, with Plates XI to XIV......... 123

New Apparatus, Reviews, etc. A Photomicrographic Outfit, by James H.

Stebbins, Jr., with Plate XV: The Pietzsch Microtome, by Edward

P. Dolby: New Model Projection Apparatus, Bausch and Lomb

Lantern “D”: The Microscopy of Vegetable Foods, by Andrew L.

Winton (John Wiley and Sons): Watson and Sons’ Instruments:

Ross Microscopes: Announcement by the Secretary ...--+++++++++++> I5I

Minutes of the Annual Meeting .........eeeee eee ce ceeeeeerereretccers 159

Treasurer's Report ..c.csccecoscrccedsancecvcoaes fem puignes/s aman maunnms 166

Custodian’s Report, Spencer-Tolles Fund ......:e eee eeeeeeeeecercerees 167

Constitution’ <chcwelsvuncccnccatevccoscecscnevbu ce owanisvnb te Same Um ummm 169

Byles fh eS ee eae een eps ae we 018 oe ate 170

List Of Metnbers ts «save cle cle vcis tence neccncce cs ae esine ss seme aes a 5s ear nee 173

List. of Subscribers cape ncbeccascessvevcss bene ens dwy wey «05's ¥u/amn umemaE 180

Tee i ee ee ale he Lhe wh ain ino ele 0. aed» pe ae 181

TRANSACTIONS

OF

The American Microscopical Society

TWENTY-EIGHTH ANNUAL MEETING, HELD AT THE LAKE

LABORATORY, SANDUSKY, OHIO, JULY 5 TO 8, 1905

THE ANNUAL ADDRESS OF THE PRESIDENT

THE RELATION OF ANIMALS TO DISEASE

By HENRY B. WARD

To the student of science who in a retrospective mood surveys

the records of the past, nothing is more striking than the failure

of medicine to attain the advancement reached in other fields. No-

where else can one find to-day among persons of education and

intelligence such a mass of fraud and superstition, such a volume of

ill-digested and undigested observations and conclusions as attach

themselves to this all-important subject and hinder its proper ad-

vance and well-balanced development. And this unfortunate condi-

tion is clearly not due to any notable difference between the con-

dition of medicine and that of other subjects in earlier days. Four

hundred years ago astronomy, chemistry and zoology were as hope-

lessly entangled in the meshes of the fabulous as was medicine, but

with the rise of science all have not advanced part passu. Whatever

the reasons for the difference, they stand on a very different basis

to-day, and, unfortunately, that subject which is of the most vital

importance for the human race has lagged in development far behind

its foundation sciences.

It is not my intention this evening to endeavor to analyze the

causes or explain the conditions which have led to this state of

6 HENRY B. WARD

affairs, but rather to discuss with you one phase of the great field

covering the etiology of disease in which the scanty and inaccurate

knowledge of the present day is especially apparent and in the

development of which the microscope plays an essential part.

The consideration of the relation of various factors to the cause

and spread of disease is of most recent origin. While popular

superstition, more often false than correct, has recorded even in

the most ancient history of medicine the source of various ailments,

it is only within the last century that there has been any critical

scientific’ study of the problem. Less than three score years cover

the epoch-making investigations of Koch, Pasteur, and their coad-

jutors, which have laid the foundations and built up the already

complex superstructure of bacteriology. By the efforts of these men

the relations of minute plant germs, unicellular organisms which we

call the bacteria, have been elucidated in great detail so as to justify

a new theory of the origin of disease and a new and successful line

of prophylaxis, or disease prevention.

Similar studies have not been made in the zoological field, but

recent discoveries seem to indicate the existence of important rela-

tions heretofore unsuspected and emphasize the hopeful character

of this new field for research. In order to secure a comprehensive

survey and place new items in their approximate position it is

fitting to review in toto the relations in which animals stand to dis-

ease, restricting the inquiry, however, for evident reasons, to such

ailments as affect mankind.

The simplest relation is manifested when the animal becomes a

carrier of disease germs. This is a purely mechanical function

and such disease-producing organisms as may be adherent to the

body of the carrier are transported unwittingly from one point to a

new environment where similar chance causes them to be deposited.

In this way such germs may be distributed widely from an originally

small focus and may be brought into inappropriate and unfortunate

relations to members of the human species. A very large number

of isolated cases might be cited to demonstrate such mechanical

transport by animal carriers. One of the best known is the trans-

port of typhoid germs by means of flies. The bacilli which are

found in excreta adhere to the feet and hairs of the fly walking

over such material, only to be carried by the next flight of the insect

to a dish of food or pan of milk left standing on a table in the

/

RELATION OF ANIMALS TO DISEASE 7

house or a bench at the door. In this new environment the germs

may multiply and with it gain entrance to the new human host with

disastrous results. Veeder has given a most vivid description of

the unsanitary conditions which actually existed in our army camps

in the Spanish-American War and which demonstrated on a large

scale this mechanical transfer of typhoid germs. Moreover, it can-

not be doubted that the bacilli actually are carried in such fashion,

for in experiments reported in the Transactions of this Society,

~ Maddox demonstrated that when such flies as have visited cultures

of disease germs, walk over sterile gelatine plates they leave foci

from which develop new colonies of bacteria of the specific sort.

Experimental evidence is wanting which shall determine the actual

extent of this infection, the distance to which such germs may be

carried, and the length of time in which they remain alive and

capable of producing an infection, as well as the other factors which

control the importance of such mechanical transport. That it does

play an important rdle can hardly be doubted, for to the numerous

instances cited in 1899 in Nuttall’s splendid monograph the inter-

vening years have added both numbers and weight. To the instance

already discussed where such transport was active must be added

the passive transfer, as of typhoid germs in oysters, which is well

established.

It should not be inferred from the preceding that only typhoid

bacilli are transported actively or passively by animal carriers. The

germs of cholera, anthrax, septicemia, pyemia, erysipelas, tubercu-

losis, and bubonic plague are said to have been transferred from one

host to another in the same way. In some cases the evidence seems

clear; in others a verdict of “not proven” must be entered; and

yet the observations already on record call for a most thorough

investigation and extended experimentation in order to reach a final

conclusion as to how widely the mechanical transfer of disease-

producing bacteria may extend. In many cases it is doubtless

purely accidental—casual—as in the hospital cases infected by flies

which Leidy records in Philadelphia, or by mosquitoes, as Giles

notes in India. In such transfer of disease germs not only are

flies the carriers, but also mosquitoes, bedbugs, fleas, and other

blood-sucking insects, though to a less extent, if present evidence

represents fairly the actual conditions. Probably such carriers of

disease will be confined chiefly to the insects and the passive agents

will be comparatively rare.

8 HENRY B. WARD

It has been observed, however, that such agents transmit disease

germs in other manner than merely adherent to the external parts

of the body. Many experiments have demonstrated that various

bacilli may pass unharmed through the intestine of the fly and be

distributed with the droppings of this insect to form centers of

development wherever they chance to be deposited. More extended

experimentation on this point is urgently needed, but one can hardly

doubt that other insects, and perhaps many invertebrates, function

in similar manner as distributors of infection. It should be noted

that this manner of distribution is not confined to bacteria alone,

although only scanty evidence is at hand concerning the mechanical

transport of other forms. Thus Grassi found that flies sucked up

with water eggs of various parasites, both tapeworms and round

worms (Taenia solium, Oxyuris, Trichuris), and that these eggs

were recovered unaltered from the dejections of the flies, while he

also caught some flies with the alimentary canal full of these eggs.

This is positive evidence that the fly is able to ingest solid bodies of

some size through the sucking proboscis. At the same time he saw

flies on his laboratory table feed on the eggs of Trichuris, and

later found the eggs in droppings deposited in the kitchen in the

story beneath, at a distance of ten meters from the place where the

insects had been feeding. Such internal transportion evidently

insures far greater freedom from damage and adverse conditions, as

well as much wider dissemination, than were the spores or eggs

merely adherent to external organs. Thus living cholera bacilli

have been voided by a fly some days after the original contamina-

tion. In the course of this period of time the fly could have wan-

dered to some distance from the place of infection.

Many investigations have shown, however, that small larvae or

adult worms like trichinae were digested by the various animals to

which they were fed, and have entirely disappeared in the course of

a few hours. Such experiments have been made with frogs, sala-

manders, land and water beetles, maggots, and earthworms. Stiles

tried some years ago a most interesting experiment which throws

much light upon this subject. He placed fly maggots with some

Ascaris lumbricoides and the latter were devoured, together with the

eggs they contained. Not only the fly larvae, but also the pupae

and the adult insects which developed from them were found to

contain eggs of the Ascaris. As the experiment was carried out in

RELATION OF ANIMALS TO DISEASE 9

very warm weather the Ascaris eggs developed rapidly and were

present in the insects in various stages. Evidently, then, the adult

fly would serve as a disseminator of the parasite, and if the eggs

obtained the proper stage of development the fly might infect man

directly by depositing them on articles of food. It is known that

certain seeds will develop only after having passed through the

intestine of birds, and it may well be that a similar biological envir-

onment is necessary in the case of some disease germs. Some such

condition would serve to explain the curious inability to infect

experimentally by direct transfer where the disease is yet readily

and abundantly transferred in nature. But the transferring insect

would not be a mere mechanical carrier; it would constitute a nec-

essary link in the life history. There are many such cases already

known, but in most of them, at least, the disease-producing organ-

ism passes through some phase in its life history in the dissemi-

nating animal, which thus becomes an intermediate host, a neces-

sary and not a casual element in the life cycle. Such forms are in

no sense mechanical carriers, and it is evident that the limits between

these two groups depend partly at least on the extent of our knowl-

edge, since a more careful investigation may show that some in-

stances of transfer which are regarded to-day as purely mechanical

involve in reality more complicated relations. It is of the greatest

importance that these relations be definitely established, for on them

depends the introduction of a rational hygiene, and yet even the

merely mechanical function of the fly in the dissemination of disease

calls for strict measures to abate this nuisance. Anyone may con-

vince himself, even by superficial observation, that both individuals

and communities through carelessness allow and produce conditions

which breed enormous numbers of unnecessary flies. Rational

hygiene calls for the removal of these conditions and the extermina-

tion of flies. Fortunately, to-day one does not need to emphasize in

civilized countries the undesirable character of bedbugs, cockroaches

and other vermin which doubtless play a part in the mechanical

transfer of disease germs, and probably are also associated more

intimately with certain maladies, as will appear in the succeeding

paragraphs.

Animals are also breeders of disease as well as carriers in a

mechanical sense; and the part they play as breeders of disease may

be either purely facultative, or, on the other hand, essential to the

fe) HENRY B. WARD

spread of the malady. Regarding the facultative rdle of animals in

breeding disease, comparatively little exact evidence is at hand. It

is somewhat generally maintained that various human diseases afflict

certain animals, and the domesticated animals which stand in such

close relations to man have been those against which up to the pres-

ent time such charges have been most frequently made. The evi-

dence is scanty, inconclusive and in some cases of no value at all;

and yet one cannot doubt that some of the germs which infect man

do live also in other animals. Even among the higher animal para-

sites but few species are confined exclusively to the human host, and

some, like Ascaris lumbricoides or Trichinella, may occur in a wide

range of hosts. It is an important duty for the students of com-

parative medicine to determine to what extent disease-producing

organisms may parasitize other hosts than man, for in this possi-

bility lies the secret of the transmission and appearance at isolated

points of new disease foci in some of the cases hitherto unex-

plained. It should be noted distinctly that when animals are facul-

tative breeders of disease they merely afford a suitable ground in

which the disease germs may multiply and an agency by which

they may be distributed. Such animals are not in any way neces-

sary to the existence or development of the germs; they only serve

to increase the percentage of infection or the area of distribution

characteristic of the disease. It is thus an important but not an

essential role. Without question it plays some part, but how weighty

its influence may be or in just what directions it may be exerted we

are at present entirely unable to measure or estimate. This is

unquestionably a most important and fruitful field for investigation.

In another sense, also, animals are breeders of disease, as when

some part of the life history of the disease-producing germs is

passed within the animal before that stage is reached in which the

germ may infect a new human host. Here the relation is an essen-

tial one, and the intermediate host is a condicio sine qua non for the

further spread of the disease. Such a relation is very widely known

among animal parasites. The embryo of the sheep liver-fluke, for

instance, must undergo certain phases of development and repro-

duction within a snail before it reaches that form which can re-infect

the sheep. The embryo of the unarmed human tapeworm must

enter another host, the beef, and grow to a bladder worm, and this

alone can produce an adult tapeworm in the human alimentary canal.

RELATION OF ANIMALS TO DISEASE It

The embryonic roundworms in the human blood must be drawn

into the stomach of the mosquito, wander out into the thoracic

muscles and grow to a definite stage of development before they

can again enter the human host and become sexually mature adults

which produce the blood-inhabiting embryos. In the case of ma-

laria, the germs of Plasmodium malariae must be drawn up into the

stomach of the Anopheles mosquito, and within the body of this

new host undergo a complicated series of changes before the new

generation of spores is ready to be injected with the saliva into the

blood of a man in whom these germs produce a new case of malaria.

Not only is the intervention of a biting insect essential, and we know

none other than the Anopheles mosquito which can ‘ fill the bill,’—if

you will allow the apparently appropriate expression,—but it is

equally true that the organism must pass through the complicated

phases of its life history in the mosquito before the latter can infect.

This is possibly still clearer in the case of yellow fever, even though

the specific organism which is the cause of the disease remains as

yet unknown. The mosquito which can transmit this disease is

also a specific type, Stegomyia fasciata, designated often as the

yellow fever mosquito. It acquires the power to transmit the dis-

ease by feeding on the blood of a yellow fever patient, but it can

infect a non-immune person only after a period of ten to twelve

days. Before that time the bite of this infected mosquito is harm-

less, and this condition can be explained only on the basis that the

organism of the disease passes through certain stages in its devel-

opment within the mosquito as a necessary preliminary to reaching

the condition in which it is able to reenter the human frame and

infect such persons as are susceptible. Until this period in the life

history of the disease germ has been completed, the mosquito re-

mains innocuous. On no other basis than this can the time interval

be explained during which the mosquito does not transmit the dis-

ease, while after that limit has been passed the insect remains

capable of infecting man up to the end of its existence, or at least

for more than two months.

The cases given illustrate in a representative way the phenomenon

of alternation of hosts as it occurs often in the life history of para-

sites belonging to different groups of animals. In some cases the

stay in the intermediate host is merely the occasion for growth and

metamorphosis as with the blood filariae in the mosquito or the

12 HENRY B. WARD

tapeworm embryo in the beef. But in other cases there is a repro-

ductive period in this intermediate host, so that the change of hosts

is associated also with alternation of generations or metagenesis.

By means of this new generation the number of spores, eggs, em-

bryos, or other infecting units is markedly increased and the com-

plicated and dangerous life cycle of the parasite finds its compen-

sating factor in multiplied numbers. Among the arthropod parasites

alternation of generations and change of hosts is rare; but among

the parasitic worms both phenomena occur frequently. Thus all

endo-parasitic flukes, so far as the life history is known, manifest

alternation of hosts and of generations; direct development has not

yet been shown to occur in any tapeworm, although there is only

rarely any new reproductive period in the life cycle. The round-

worms, or Nematoda, display every grade from the most extreme

simplicity and directness of development and transfer, to such com-

plicated changes and wanderings as have even yet eluded the scru-

tiny of the closest investigator or when announced have aroused

the ridicule of the scientific world on account of their improbability.

As an excellent instance of these complicated relations may be cited

the life history of the European hookworm, published by Looss,

little more than a year ago. Looss has followed the migration step

by step from the time the minute larvae penetrate the hair follicles

of the skin, enter a lymph space or a capillary, to be carried by the

current through the vessels ultimately into the right side of the

heart and from there into the lungs, where they desert the vascular

system and migrate into the air cells, and then wandering outward

along bronchioles, bronchi, and trachea pass over the dorsal margin

of the larynx and into the oesophagus, from which their pathway

lies directly back through the alimentary canal to their final location

in the small intestine. This migration requires ten weeks, during

which time they pass through moults and grow in size, attaining —

the adult form and sexual maturity only after arrival at the end of

the journey. Here the entire life cycle is passed in a single host,

but its different phases are associated with various organs. In still

other cases among the Nematoda a free-living generation alternates

with the parasitic generation, instead of two, which are found in

different hosts. |

Concerning conditions among the Protozoa, there is less definite

knowledge of the life history than among the higher groups, but

|

|

5

RELATION OF ANIMALS TO DISEASE 13

instances of all the conditions cited for the worms may also be

found here. Some species undergo direct development, others

make a single or even a double change of hosts, and in some two

generations of different type alternate in the complete life cycle

of the organism. Thus the amoeba of tropical dysentery (Entamoeba

histolytica) seems to develop directly ; the blood amoeba of malaria

(Plasmodium malariae) goes through an asexual reproductive

cycle in man, and another, the sexual cycle, in the mosquito. In

this case we know that the mosquito is not the mere mechanical

carrier of the disease germ, but that it is a necessary link in the

life history, a breeder as well as a transmitter of disease. Re-

garding the rdle of the cattle tick in Texas fever, it may be inferred

with great probability that it plays a similar part, even though

the history of the parasite within the tick has not yet been worked

out. In other diseases, such as sleeping sickness, where the para-

site, a flagellate protozoan known as a trypanosome, is transmitted

by a biting fly, familiarly called the tse-tse fly, there is less evidence

on which to base a conclusion. The tse-tse fly may be purely

mechanical in its intervention; it seems more probable, however,

that it plays a more intimate part. The instance shows very clearly,

however, that until the life history has been elucidated, it is impos-

sible to determine the relative importance of any element in the

series, or intelligently to combat the disease which evidently should

be attacked at its weakest point. This factor will be considered

more in detail later on.

But animals also stand in a causal relation to disease; certain

forms are definitely shown to be producers of disease and in

this aspect demand especial consideration. This fact has been gen-

erally recognized in the case of a few parasites from the earliest

days of medical history. The fiery serpent of the wilderness was

no doubt the guinea worm, of which the most ancient medical

writings make note; and in this instance not only the cause of

the malady, but also the general mode of infection through drinking

water, and the method of cure, the removal of the worm, were

known to the Egyptian as well as to the Greek physicians. But

such instances are rare. Regarding merely even the larger, more

conspicuous parasites of man the wildest ideas were formerly current

as to their origin and their effect on the system. Thus tape-

worms were supposed to originate from thickened mucus, or from

14 HENRY B. WARD

an abnormal condition of the alimentary canal; and various animal

parasites were from time to time regarded as the causes of cholera,

typhoid, and other similar diseases. Such views as these prevailed

generally even less than a century ago, and it is not strange if in

consequence of more accurate knowledge on these points and of the

rejection of such wild theories of disease, the pendulum has swung

to the opposite extreme and animal parasites have come to be con-

sidered of insignificant importance in the etiology of disease.

Two factors tended to strengthen this view and further belittle

the possible role of animals as disease producers. In the first

place, with the possible exception of malaria, no animal organism

was known to be the cause of any general disease; and while the

animal nature of the Plasmodium malariae was never doubted in

any considerable circle, the case stood so evidently isolated that

it emphasized all the more its own peculiarity. But even more

powerful than this was the rise of a new science, bacteriology.

Certain minute plant germs had been found to be the cause of decay,

why not of disease? In response to the needs of the case there

arose a new technique for handling and studying these forms, a

rigorously analyzed series of conditions for determining their pos-

sible relation to disease; and a new field of science was organized.

Discoveries followed one another with marvelous rapidity and every

year saw the elucidation of the cause of new maladies. It seemed

as if the secrets of disease had been laid bare; men had traced the

causes to bacteria in many cases with such success that they con-

tinued to follow the same line in other yet unexplained diseases,

confident that there was only some minor defect in technique which

would soon be overcome and the solution obtained. Indeed, the

very name “ disease germs” was regarded as equivalent to bacteria.

There is no doubt that success in this direction served to draw

attention away from the signs which presented themselves in other

fields and particularly to minimize the animal organism as a causal

factor in disease. Recent discoveries of great import which have

crowded hard upon each other, are disclosing here a new field and

stimulating the investigation of neglected territory. Let us now

examine sertatim the different groups of animals to secure a clear

idea of the role played by each in the production of disease.

The disease-producing organism works slowly, insidiously, saps

the vigor of the infected individual without consuming the substance

RELATION OF ANIMALS TO DISEASE 15

so as to destroy life by immediate destruction of the body. It

is clearly not carnivorous, but rather parasitic in habit; con-

sequently among the vertebrates, as well as among the largest and

most powerful invertebrates, one could not expect to find such

forms. These largest species might be carriers of disease or even

breeders of sickness, but they could not constitute the immediate

cause of the malady. It may be interesting to note in passing an

apparent exception to this rule. The lamprey attaches itself to other

fish and is directly the cause of the ulcers on the skin which mark

the points of the lamprey’s fixation, and of the anemia which fol-

lows its blood-sucking and often induces the death of its host. But

this instance stands alone.

In the great majority of cases the disease producers are small

organisms, or at least gain their entrance into the body of the host

in a form so minute as to defy detection. The arthropods furnish

many carriers of disease and breeders also, as the extended refer-

ences already made to these forms suffice to show. The more or

less perfectly acquired external parasitism of these forms is admir-

ably adapted to these functions, but such animals are not the imme-

diate cause of disease, and when sickness follows a bite of a fly, a

spider, or a tick, the effect is more probably due secondarily to the

bite and primarily to some other organisms introduced thereby.

In the case of the parasitic worms the conditions are decidedly

changed. Here are species which parasitize within the body, often

suck the blood of the host, lacerate delicate mucous membranes,

induce internal hemorrhages, in some instances feed upon the cells

of the tissues, and destroy important organs, or grow to such a

size as to encroach upon normal structures and functions. In addi-

tion to these anatomical interferences, some of the parasitic worms

are known to produce waste matter in their own biological processes,

toxins, which act deleteriously upon the host organism and evoke

abnormal and serious symptoms in it. Thus Vaullegeard has iso-

lated experimentally from tapeworms two chemical substances

which act upon the blood and nerve and when injected into experi-

mental animals subcutaneously produce the epileptic symptoms that

characterize severe cases of tapeworm infection. Then the physician

speaks of a “ bothriocephalus anemia” recognizing a definite group

of symptoms, a distinct disease produced by the parasitism in man

of the broad fish tapeworm (Dibothriocephalus latus). Here the

16 HENRY B. WARD

animal is the immediate cause of the disease and the removal of

the tapeworm is followed at once by the disappearance of the unde-

sirable symptoms.

While there are some animal parasites which are believed to be

harmless, or, better expressed, do not do any damage to the human

system, so far as present knowledge extends, yet the studies of

recent years have furnished constantly increasing evidence of the

pathogenic réle of these organisms. They do damage indirectly by

irritating the delicate mucous membranes and by lacerating them,

thus giving access to the omnipresent bacteria, a danger which has

been greatly underrated. But they are also the direct cause of

disease which in consequence of their part in its production the

physician names after the species of parasite, as trichinosis, uncinar-

iasis, hydatid disease, etc. Note further that they are not factors

of trivial importance in general hygiene or of little bearing upon

the welfare of a nation as a whole, and that a large percentage of

such diseases can be treated only by preventive medicine. Thus

trichinosis, which is caused by eating pork containing living trichinae

(Trichinella spiralis), is accompanied by a high mortality and even

yet is a serious disease in northern Germany; its prophylaxis 1s,

however, exceedingly simple and no one who is careful to avoid

under-done pork will ever suffer from its attacks. Again, hook-

worm disease, or uncinariasis, has been shown by the researches of

Stiles and others to be very abundant in certain parts of our

southern states. The presence in the alimentary canal of myriads

of minute hemorrhages caused by the action of these worms, results

in a chronic anemia which prevents the attainment of physical or

mental development, stunting the individual and leaving him on

arrival at years of maturity little more than a child in body and

in intellect. Much of the degeneracy of the poor white trash of the

South is due not to inherited defects or to family shortcomings so

much as to the presence of this parasite which from early childhood

has continually sapped the vitality of the individual. It needs no ex-

tended argument to demonstrate the sociological effect of the rec-

ognition and removal of this one cause of disease. Nor will anyone

doubt the desirability—yes, the necessity—for a careful investi-

gation into the life history and effects of these parasites. For

from the life cycle is to be obtained the clue to the means of

attack, to the weak spot in the armor of the disease, on which its

RELATION OF ANIMALS TO DISEASE 17

ultimate destruction depends; and everyone recognizes as the ulti-

mate goal of medicine as a science, the eradication of such diseases,

that the physical man may move forward toward the possibilities

in perfect development with which he is endowed.

That which I have outlined has been known in part for many

decades, even though the investigations of recent years have con-

tributed much towards a clearer comprehension of the question.

Among. the Protozoa, however, the last few years, and even months,

have brought discoveries of the most startling character regarding

their relation to disease. It was in 1880 that Laveran first discov-

ered the amoeboid parasite in the red blood cells now universally

recognized as the cause of malaria, and not until 1899 was its life

history clearly outlined, while even yet some minor details of the

picture are lacking. Since the opening of the new century there

has come the demonstration of the cause of sleeping sickness, a

terrible disease of tropical Africa, in a flagellate protozoon (Trypan-

osoma), other species of which in the blood of various domestic

animals have been shown to give rise to widespread and fatal epi-

demics in other countries; the parasite of smallpox has been found

to belong to this same group and its life history has been deter-

mined partly at least. The disease known as kala-azar, dum-dum

fever, or splenomegaly, a fatal malaria-like malady of India and

Africa, has been traced to another protozoan parasite; in yellow

fever it seems probable that such organisms are the exciting cause;

in various other diseases they have been seen, even though in some

cases subsequent investigation has failed to demonstrate the para-

sites and confirm the reports; and finally within this year accounts

by well-known German investigators proclaim the discovery of

the cause of syphilis in a hitherto undiscovered protozoon of the

order of flagellates. In all of these maladies the bacteriologists

have been searching with great care for the etiological factors, but

their efforts have been fruitless. It is apparent that the new field

will demand its own technique, and until that has been developed

and the proper standards of judgment formulated, much work will

necessarily go to waste and many errors be committed.

These organisms, the unicellular animals, are distinctly analogous

to the unicellular plants, among which the bacteria stand as the

characteristic disease producers. Indeed, the recent studies have

shown that one genus, Spirochaeta, long known and hitherto clas-

18 HENRY B. WARD

sified among the bacteria, is probably not such, but rather a flagel-

late protozoon. And possibly other genera of Protozoa are also

wrongfully assigned to the bacteria. On the other hand, zoologists

have long recognized certain forms of Protozoa as pathogenic, pro-

ducing disease among the various other animals, and this is at least

an indication of their filling a similar role in the human body. |

In consideration of these facts, it is not unreasonable to believe

that we stand now at the opening of a new field which is to make

of itself in the future what bacteriology has made in the last half

century. There is need of a Pasteur, a Koch and their confréres

to lay the foundations strongly and to analyze with equal sharpness

the relation of these animal micro-organisms to disease. Even now

the new field has been recognized and the London School of Trop-

ical Medicine has appointed this year an investigator in protozo-

ology—however unfortunate the form of the term may be.

There are already listed more than thirty of the Protozoa which

parasitize the human body. Regarding many of them our knowledge

is exceedingly scanty, but of others it may be affirmed definitely that

they are the cause of diseases which rank among the most dangerous

of those to which man is subject. Among these forms I have

included only those that are distinctly recognizable in structure as

Protozoa, though their life histories and exact relationships are yet

unexplained; but beyond these limits lies a vast horde of uniden-

tified structures, interpreted by some observers as parasitic Pro-

tozoa, but regarded by others as parasitic fungi, and by still others

as products of cellular degeneration or other pathological changes.

Such are the cancer parasites of several investigators, the organisms

of leukemia, scarlet fever, and other diseases. No doubt some of

these will be shown by further research to be in fact independent

organisms of parasitic habit and the cause of disease, and it seems

probable that many of them will fall within the group of Protozoa, —

the unicellular animals. Here has been opened up a new field in

which the microscope is the essential instrument of investigation.

All the work to be done in it depends upon this instrument, without

which the very existence of these organisms would have remained

unsuspected. Following close upon the wonderful discoveries of

the histologist, the pathologist, the bacteriologist, and the clinician,

these studies furnish new evidence of the supreme importance of

the microscope in the development of scientific medicine, in the

attainment and preservation of the health of mankind.

RELATION OF ANIMALS TO DISEASE 19

There is left but a paragraph in which to mention another aspect

of the subject of this address. Even under the narrow limits of

the topic—the relations of animals to disease—there is one phase

which in justice to them should not be entirely omitted. Animals

stand also distinctly as preventers of disease; and this in the first

place as destroyers of disease germs. Among the Protozoa, which

have already been exploited as the producers of disease, are found

also the organisms which play the most important part in the puri-

fication of sewage-contaminated streams by consuming the bacteria.

These forms are specifically ciliates, of which the common slipper-

animalcule (Paramecium) may serve as a typical form; they abound

in all waters, especially in those containing decaying matter, and

devour countless numbers of bacteria. Through their activity it

becomes possible for one city to drink the diluted sewage of another

city higher up on the watershed without losing all its citizens from

intestinal diseases.

Modern science has also made use of animals in combating dis-

ease; as producers of antidotes, either in the form of cOwpox or

-vaccine, or in the rdle of test animals and of serum producers

manufacturing antitoxins of various sorts; many animals discharge

in this way a most essential function in modern life. But the dis-

cussion of this phase lies beyond the demands of the present occasion.

In closing, let me call your attention to the bearing these studies

on the relations of animals to disease have on the science of medi-

cine. Any rational method of cure depends upon the distinct rec-

ognition of the cause of the malady. Any other basis gives unlim-

ited opportunity for chicanery and fraud and for the despoilation

of the people in the name of medicine so general at the present time.

But more than that, preventive medicine is to be the ultimate product

of the scientific studies of to-day; no one can question that it is

a far higher and more desirable type than curative medicine which

now generally seeks to remedy the ills begotten through ignorance.

The loss to the world by preventable disease is enormous ; it includes

many of the wise and the good, of the best products of human evolu-

tion during past centuries, for no selective action determines that

the worse element shall be wiped out. In truth, the delicate ner-

vous balance of the highly developed human organism seems to be

more easily disturbed by the attacks of disease than the grosser

clay in which all energy has gone to physical development. To stop

20 HENRY B. WARD

this loss is the greatest problem of the future in medicine. And the

very first step in this problem is the positive determination of causes

of disease and of the means by which they are transmitted and mul-

tiplied. Without this knowledge rational prophylaxis is impossible ;

before it and the results of associated investigations of purely scien-

tific character quackery must yield as the night before the day,

schools and theories will disappear, and medicine will take its right-

ful place among the sciences.

A CONTRIBUTION TO THE MICROSCOPIC STUDY OF

PEN AND INK LINES

By MARSHALL D. EWELL

In the third edition of a work entitled ‘‘ Bibliotics, or the Study

of Documents,” etc., by Dr. Persifer Frazer, the learned author has

devoted an entire chapter (Chapter X) to the study of so-called

serrations in ink and lead-pencil lines and has attempted to show

that by means of these serrations the authorship of a disputed

writing may be determined. The author in the chapter mentioned

calls these serrations “ provisionally, variations of nerve force,” and

states that “ the fact of the existence of these peculiarities is unques-

tioned and the value of the observations where proofs of identity or

non-identity are sought is unquestionable.” For a full presentation

of the author’s views reference should be made to Chapter X of

his work where a clear presentation of the theory profusely illus-

trated will be found.

The fact that the views of the author have, without, as it seems,

any independent investigation, received a quasi indorsement from

Drs. Wood, Mitchell, and Witmer, as appears from their letters pub-

lished in the preface and body of the work, and also from Winslow

in his recent work (1905) on ‘“ Applied Microscopy,” page 139,

together with the further fact that Dr. Frazer has in at least one

case! attempted by this method to establish the authorship of a dis-

puted writing, demands a careful investigation of the theory before

it is allowed any weight in jeopardizing human life or liberty.

That numerous irregularities in the contour of lines traced on

paper by a pen or pencil exist will not be denied. The grosser ones

are apparent to the naked eye and may be due to inexperience of

the writer or to an unstable condition of his nervous system, however

caused.

A microscopic examination of ink and pencil lines will also dis-

1The State of New York vs. Kennedy, tried in the City of New York in

Igol.

22 MARSHALL D. EWELL

close the existence of more minute irregularities or “ serrations,” if

one prefers that term; but that they have any significance or weight

whatever in determining the authorship of a disputed writing is

denied.

In order to settle this question the writer made a series of obser-

vations early in the present year which seem to him to bear directly

upon this question and to show that the claim has no reasonable

grounds to warrant its acceptance. The process of exclusion of the

various factors involved was the method of research adopted.

Granted the existence of the so-called serrations, what is their cause?

That they have in written characters any regularity of sequence

is denied, and in the writer’s judgment such regularity is not capable

of proof. In the use of a diamond tool in ruling micrometers on

glass or metal, or in turning metal in a lathe with a metal tool, if the

tool is not properly held or ground, it frequently happens that it

will chatter and produce regular serrations upon the glass or metal

surface upon which the work is performed. These serrations may

have rounded extremities or may be sharp and regular like saw-

teeth; or the surface may be irregularly cut or torn. Let anyone

who is the possessor of an Abbé test-plate examine it and he will

have a good example.

A microscopical examination of the engine-ruled lines on writing

paper will also disclose these so-called serrations in great numbers.

In the investigation of the question the writer first ruled lines on

paper with a pen and ink by means of a dividing engine drawing

the ruling carriage by a string in order to cut off any personal ner-

vous influence. The lines so drawn uniformly exhibited serrations.

The writer had years previously ruled lines on glass and through

metal films on glass and knew from experience thus gained that

serrations might be produced by a ruling tool improperly mounted ;

but that if the tool was properly held, a line could be produced that

showed not the slightest irregularity of contour under a power of

1500 diameters.

The next step in the investigation was to write with a pen not

charged with ink on a thin film on glass in the same manner that

ordinary writing is produced, the only difference being that the lines

were not produced on paper by ink. Some experiments were nec-

essary in order to produce a film that would exactly record the

movements of the pen perpendicular to and across the surface oper-

MICROSCOPIC STUDY OF PEN AND INK LINES 23

ated upon. Plate glass was held over a smoky flame and a very

thin film of carbon thereby deposited upon its surface. This made

a good medium for recording the action of the pen. The film must,

however, be very thin, as otherwise the carbon will gather on the

point of the pen and the accumulation will be deposited irregularly

along the edges of the line much as a snow plough deposits the

snow along the sides of the track, the difference being principally

one of degree. Another film was produced by flowing the glass

with a thin solution of wax and asphaltum in benzole. If too

thick a film is used the same phenomenon of the deposition of the

substance of the film along the edges of the line will be manifested.

Satisfactory films having finally been made, thin enough to record

the pen movements, they were written upon by various persons

under the writer’s direction with an ordinary dry steel pen held and

operated in the usual manner, and in every instance the margins

of the lines under a power of over one hundred and twenty diam-

eters were absolutely clear, sharp and free from serrations of any

sort. The ordinary writing of the same persons on paper with

pen and ink had previously shown abundant serrations.

A microscopic examination of ordinary writing paper will uni-

formly show irregularities of surface, consisting of furrows, pits,

and elevations. This fact was ascertained by a microscopical ex-

amination of the surface of the paper held at right angles to the

optic axis of the microscope and from the examination of sections

of the paper, the surface being parallel to the optic axis. These

pits, furrows, and elevations afford an ample reason for the exist-

ence of the serrations in question, and the experiments above re-

corded show that they are not due to variations in the nerve force

of the writer, but to irregularities in the surface of the paper itself.

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ON THE OCCURRENCE OF TRYPANOSOMES IN THE

BLOOD OF RANA CLAMATA

By JAMES H. STEBBINS, Jr.

WITH ONE PLATE

Some time ago, while investigating the subject of haemosporidia

in the blood of frogs, I accidentally came across a spring frog (Rana

clamata) which was not only abundantly infected with haemospor-

idia, but was also infected with another form of parasite which upon

further investigation was found to belong to the class Monanida, to

the family Trypanosomidae, and to the genus Trypanosoma. The

trypanosomes found in this frog’s blood differed so radically from

any with which I was acquainted that it was thought a short account

of the various forms of organisms observed might be of interest.

Although much has been written upon the morphology of the

trypanosomes of warm-blooded animals, very little on the other hand

has been said concerning the trypanosomes of cold-blooded animals,

and the one receiving the greatest amount of attention at the hands

of scientists is the Trypanosoma rotatorium, found in the blood of

the European edible frog, Rana esculenta, and to some extent also

in the blood of Rana temporaria and Hyla arborea.

This trypanosome is described by Stiles (Emergency Report on

Surra; Washington, 1902, p. 39) as follows:

“Trypanosoma.—4o-80 microns long, by 5-10 microns broad.

Flagellum 10-12 microns. Body compressed, semilunate, twisted.

The convex border membranous and undulating; the posterior ex-

tremity of the body portion pointed, curved inward; the opposite

one produced into a long tag or tail-like appendage which almost

equals in length the remainder of the body; surface of the body

coarsely striate longitudinally; endoplasm or parenchyma slightly

granular, endoplast ovate, central.”

The trypanosomes observed in the blood of Rana clamata were of

various shapes, depending upon which stage of their life cycle they

had reached, but among these, two forms seemed to predominate,

which, briefly described, are as follows:

26 JAMES H. STEBBINS, JR.

The smaller of these (pl. 1, fig. 1), which I name Trypanosoma

clamatae, in reference to the frog in which is was found, is long,

slender, and pointed at both ends, and looks very much like an eel

while gliding through the blood plasma. It propels itself with a

peculiar jerky, wriggling motion, and with sufficient force to easily

push aside any blood corpuscles which may happen to be in its path.

The anterior end of the parasite bears a long flagellum which is

moved so rapidly that it is almost impossible to see it while the

parasite is in active motion. The posterior end is extended into a

sharp point. The dorsal side of the body is surmounted by an

undulating membrane which extends from a point very close to the

centrosome to the anterior end of the body. A large oblong nucleus

is found on the ventral side of the body, not very far from the

anterior end. A large centrosome may be seen at a point almost

one-third the way from the posterior end, and occupying nearly the

full width of the body.

The flagellum starts at a point very close to the centrosome,

follows the outer edge of the undulating membrane till the ante-

rior end of the body is reached, and then becomes free. The body

cytoplasm is only slightly granular, and stains easily with polychrom

methylen blue and eosin, the Wright stain giving the best results.

The body length is about 21 microns, and the diameter 2.5-2.8

microns. The flagellum is 12-13 microns long, thus making the

total length of the body and flagellum 33.7-41.7 microns. This

organism is the full-grown, or mature trypanosome.

The larger of the two parasites (pl. 1, figs. 4 and 5), which I

have already referred to, when observed in the fresh unstained

blood, appears of a light greenish color; the body is as a rule semi-

lunate when in a quiescent state, but when in active motion it may

assume a great variety of grotesque shapes (pl. 1, fig. 5). The

posterior end is pointed, and extends into a tail-like appendage

which looks almost like a second flagellum. The convex side or

back of the parasite is surmounted by a large undulating membrane,

which is kept in constant very rapid motion. The anterior end of

the body is provided with a long, slender flagellum, which is like-

wise kept in constant whip-like motion. The flagellum starts at a

point near the centrosome, and extends along the outer edge of the

undulating membrane to the anterior end of the body, where it

becomes free. The ectoplasm is coarsely granular, while the endo-

TRYPANOSOMES IN BLOOD OF RANA CLAMATA 27

plasm is finely granular. A large spherical nucleus is usually found

at about the center of the body, but nearer the undulating membrane

than the opposite side. The body is also provided with a good-

sized centrosome which is located close to the convex side of the

parasite, about half way between the nucleus and the posterior end

of the body. The body length is 27.56-47 microns, and the diam-

eter at its widest part 16.78-28.51 microns; average length 35.74

microns. The length of the anterior flagellum is 5.96-14.79 mi-

crons, the average 10.37 microns. The length of the tail-like ap-

pendage is 3.07—4.47 microns, the average 3.77 microns. Therefore,

the total length of the parasite, including tail and flagellum, is

44.18 microns. The parasite just described is one of the division

forms of Trypanosoma clamatae.

I am unable to discover that any attempt has so far been made

to work out the life history of the trypanosomes of frogs, and per-

sonally I have been no more successful than those who have pre-

ceded me in this line of work, but by analogy with Trypanosoma

lewisi, found in the blood of rats, and whose morphology has been

pretty clearly worked out, we know that multiplication of the species

may occur by either longitudinal or transverse division, or by

segmentation. .

In the case of Trypanosoma clamatae, I have found that multi-

plication of the species also takes place by both longitudinal and

transverse division, but I have so far failed to observe any signs of

multiplication by segmentation.

According to Rabinowitsch and Kempner, after a study of many

stained and unstained preparations of Trypanosoma lewisi, the cen-

trosome and nucleus of these flagellates is an interdependent whole

which corresponds to the nucleus of the other flagellates. They

assume that in the early stages of development of the parasite the

centrosome and nucleus represent a whole which, as the parasite

matures, splits up into two parts which pass to either end of the

animal. ‘They also consider the nucleus of the trypanosome as being

made up of two parts more or less separated ; the small spot situated

in the posterior end of the parasite they consider to be a nucleolus,

while the larger structure, situated in the front end, they call the

chromatin heap. )

Wasielewski and Senn, on the other hand, consider that the cen-

trosome is in no way connected with the nucleus. According to

their ideas, the trypanosome is made up of two parts, the “ plasma”

28 JAMES H. STEBBINS, JR.

and the “periplast.” The “plasma” is the body of the parasite

containing the nucleus. The “periplast” is the outer covering of

the parasite and includes the centrosome, flagellum, undulating

membrane, and the outer coat investing the body of the organism.

According to this hypothesis the centrosome is intimately connected

with the undulating membrane, and has nothing to do with Rabino-

witsch and Kempner’s nucleolus, nor the micro-nucleolus of Plim-

mer and Bradford.

In multiplication by transverse division in the case of Trypano-

soma lewisi the first thing to be observed is a change in the out-

lines of the trypanosome. The sharp beak becomes more blunt and

the flagellate end becomes rounded ; the long, slender body becomes

thickened and swollen with a decided increase in size. There is a

multiplication of both nuclei and centrosomes which are arranged

in a line parallel to the longitudinal axis of the parent trypanosome.

The nuclei and centrosomes now divide, and new flagella are

formed, each of which is derived from a centrosome and emerges

from the parent parasite on the side which bears the undulating

membrane; after a while these attain full length, while the old

flagellum becomes destroyed and finally disappears. There is also

division of the protoplasm of the parent trypanosome along lines at

right angles to the longitudinal axis, so that each new segment of

protoplasm is supplied with a nucleus, a centrosome, and a flagellum.

The daughter cells soon become liberated in the blood and may be

recognized by their small size, but they gradually lengthen out into

the characteristic mature organism.

Multiplication by longitudinal division bears a certain analogy to

that of transverse division, there being the same change in outline

and the same multiplication of nuclei and centrosomes, but the

arrangement is in a transverse line parallel to the longitudinal axis.

The new flagella make their appearance at the anterior end of the

parent parasite, are closely arranged about the old flagellum, and

correspond in number to the centrosomes.

The foregoing account of the multiplication of Trypanosoma

lewisi closely resembles what I have observed in the case of Trypa-

nosoma clamatae, though I have not been so fortunate as to view

the latter parasite in all its phases of division. From what I have

seen, however, I feel confident that multiplication occurs by both

longitudinal and transverse division substantially as above set forth.

In what is to follow an attempt will be made to illustrate two

TRYPANOSOMES IN BLOOD OF RANA CLAMATA 29

phases in the multiplication of Trypanosoma clamatae: (1) The

swelling and increase in size of the organism prior to division ; (2)

The multiplication of the nucleus and centrosome, and the longi-

tudinal and transverse division of the body cytoplasm.

In pl. 1, fig. 2, we have a parasite preparing to divide. When

compared with the mature parasite in figure I, it will be seen

that the former has considerably increased in size, both longi-

tudinally and transversely, but as yet it shows no signs of division.

In figure 3, we have a trypanosome in a little more advanced

stage of multiplication as may be shown by the increased longi-

tudinal and transverse size of the parasite. The nucleus also

appears to have divided. In pl. 1, figs. 4 and 5, the two parasites

have become so greatly enlarged that it may be fair to assume that

they are just on the eve of division, though no actual signs of the

multiplication of nuclei and centrosomes, or division of the body

protoplasm, are visible. ‘The one in fig. 5 is of particular interest ;

it represents a parasite of peculiar shape preparing to undergo

division ; the body is greatly swollen and the centrosome is located

nearly in the center of it, while the nucleus is either absent or

invisible. In pl. 1, fig. 6, we have a trypanosome in a still more

advanced state of division; the body is greatly swollen and enlarged ;

the nucleus is also greatly enlarged, and evidently preparing to

divide; the centrosome, however, shows no signs of multiplication,

while the body cytoplasm shows distinct evidence of longitudinal

cleavage. In pl. 1, fig. 7, the parasite shown is somewhat similar

to that of fig. 8, but in this case the centrosome is clearly seen to

be double while the nucleus is very indistinct; the body cytoplasm

also shows signs of longitudinal cleavage. In pl. 1, fig. 8, we

have a rather peculiar form of division and one which is somewhat

difficult to understand. The body of the parasite, as in the preced-

ing illustration, is greatly swollen ; the centrosome is intact, but the

nucleus seems to have divided. The body cytoplasm clearly shows

transverse cleavage, one segment of which, carrying the undulating

membrane and flagellum, is probably just about to be split off from

the parent trypanosome to form a daughter parasite.

In the foregoing I have tried to show how multiplication in

Trypanosoma clamatae probably takes place, but, owing to lack of

material have been unable to follow the various stages of division

to completion. It is to be hoped, however, that further investi-

gation will throw more light upon the subject.

30 JAMES H. STEBBINS, JR.

EXPLANATION OF PLATE

Plate I

Photomicrographs illustrating the characters of Trypanosoma clamatae

nov. sp. Magnified 667 diameters. For explanation see the text.

PisAd es |

m

i.

i

oe

TAPEWORM CYSTS (DITHYRIDIUM CYNOCEPHALI N.

SP.) IN THE MUSCLES OF A MARSUPIAL WOLF

(THYLACINUS CYNOCEPHALUS)

By B. H. RANSOM

WITH FIGURE

At the autopsy of a captive marsupial wolf (Thylacinus cyno-

cephalus), which died in November, 1904, at the National Zoological

Park, Washington, D. C., where it had been kept since September

3, 1902, the voluntary muscles and heart were found by Dr. A. Has-

sall heavily infested with tapeworm cysts (specimens No. 3888 and

4060, Helminthological Collection, Bureau of Animal Industry).

These cysts, whitish in color, oval, measure I to 2 mm. in diam-

eter. The wall of the cyst, 50 to 150,» in thickness, is composed

of fibrous connective tissue. At each pole, amid the muscle fibers

in which the cyst is imbedded, there is a deposit of fat globules

as commonly occurs in connection with the cysts of Trichinella.

The larval tapeworm almost entirely fills the cavity of the cyst. It

is covered with a cuticle about 5,» thick. The parenchymatous

tissue composing the bulk of the body contains numerous calcareous

corpuscles. A caudal vesicle is lacking. The scolex in all of the

specimens examined was completely invaginated. It is furnished

with four well developed suckers 150 to 200 in diameter and bears

no trace of a rostellum. The excretory canals open into a tube-like

invagination at the posterior end of the body, lined with cuticula

and measuring 150 to 200 in depth and about 25 in diameter.

A number of similar larval forms have heretofore been reported

from lizards, snakes, monkeys, field-mice, rabbits, pole-cats, mar-

tens, weasels, birds, cats, dogs, foxes, ichneumons, and rats, and

have been grouped together in the genus Dithyridium Rudolphi,

1819, the name of which was improperly changed by Diesing (1850)

to Piestocystis. In most cases the worms have been found in rela-

tion with the body cavity, its membranes, or contained viscera, and,

except in the present instance, have not been reported in the volun-.

tary muscles nor in the heart.

32 B. H. RANSOM

The life history of none of the species included in Dithyridium is

known. It seems, however, safe to infer that these forms repre-

sent intermediate stages of tapeworms with four suckers and with-

out rostellum, occurring when adult in carnivorous animals. The

tapeworms of the genus Mesocestoides fulfill these requirements,

and Neumann, who was probably the first to suggest a connection

between Dithyridium and Mesocestoides, has recorded (1896) some

observations in support of the view that Dithyridium. bailleti, occur-

ring in dogs and cats, is a larval stage of Mesocestoides lineatus,

which occurs in the same hosts. The results of some feeding

experiments which Neumann performed, although not conclusive,

tend to show that he was on the right track, and offer encourage-

ment for further investigations along the same lines.

If Neumann is correct in his opinion as to the identity of Meso-

cestoides lineatus and Dithyridium bailleti, it may be inferred on

the basis of analogy that the tapeworm cysts found in the marsupial

wolf are intermediate stages of tapeworms belonging to Mesoces-

toides, or a nearly related genus. Although it is not impossible that

the wolf may have been already infested when received at the

Zoological Park, it seems more likely, since none of the cysts showed

degenerative changes as would be expected if the infection were of

old standing, that the infection was comparatively recent and the

chances favor the probability that the wolf became infected from

swallowing the eggs of tapeworms harbored by some of the neigh-

boring animals at the Park.

As a name for these larval forms I would propose Dithyridium

cynocephalh.

EXPLANATION OF FIGURE

Plate II

Fig. A. Dithyridium cynocephali encysted in muscle of marsupial wolf.

X 35:

|

:

PROBSTMAYRIA VIVIPARA (PROBSTMAYR, 186s)

RANSOM, 1907, A NEMATODE OF HORSES

HERETOFORE UNREPORTED FROM

THE UNITED STATES

By B. H. RANSOM

WITH ONE PLATE

Among the numerous species of roundworms which infest the

large intestine of the horse there is one species which is notable

on account of its very small size and the enormous numbers in

which is may occur.

This species was first described by a German writer, Probstmayr

(1865), under the name of Oxyuris vivipara. That author exam-

ined the caeca of fifty horses, and in a few cases the remainder of

the large intestine, during the months from November to May, and

found the worms in question present in seven caeca out of the

entire number examined. No specimens of the male sex were

encountered. The females measured 2.45 mm. long by 0.11 mm.

wide at the level of the bulb of the esophagus. The distance from

the anal opening to the tip of the slender-pointed tail was 0.86 mm.

Three stages in the development of the females were noted, namely,

immature females, pregnant females, and females which had given

birth to young. The pregnant females contained two embryos

which could be forced out of the vulva by pressure. These em-

bryos measured 1.48 to 1.59 mm. long by 0.04 to 0.10 mm. wide,

and were completely developed with the exception of the genital

organs.

Perroncito (1882: 337) noted the presence of this species in

Italy, having found millions of individuals in the large intestine

of a horse. He also makes reference to it in a later work (1886:

300-301) and considers that it should be placed more properly in

one of the two genera, Rhabdonema or Anguillula.

Fiorentini (1890: 21-25), who met with this species in two out

34 B, H. RANSOM

of a number of horses examined by him in Italy, has given, with

the exception of a few slight misinterpretations and omissions, a

fairly accurate and complete description based on specimens of the

female.

Railliet (1893: 549) gives a short description based on the

accounts of previous authors and assigns the worm definitely to the

genus Angutllula.

The very small transparent Oxyuris from the large intestine of

an Egyptian donkey, mentioned by Looss (1902: 119), possibly

belongs to the species under discussion.

Jerke (1902: 113-127), in post-mortem examinations of thirty-

eight horses and asses found the parasite in fifteen cases, distributed

throughout the entire large intestine. More fortunate than earlier

observers, he discovered the male, and in two cases found it in

rather large numbers, in the proportion of one male to about twenty-

fve females. He gives the maximum length of the full-grown

female as 2.9 mm. with a width of 0.095 mm. ; the male measured

2.6 mm. in length by 0.084 mm. in width. According to his inter-

pretation the mouth is surrounded by three lips. The spicules

Jerke describes as equal, measuring 0.072 mm. in length by 0.0075

mm. in average width. He saw no papillae on the tail of the male.

In addition to a number of pages of anatomical description with

figures he makes a few remarks in regard to the life history and

considers that infection takes place from the ingestion of food or

water which has been contaminated by feces from infected animals.

In support of this view he mentions the fact that the worms remain

alive in manure for four or five days after deposition. Having

gained entrance to the body the worms are able to multiply indefi-

nitely, completing their entire life cycle within the body and thus

differing from most parasitic nematodes which at some stage must

pass to the exterior and again re-enter a host before their develop-

ment is finished.

Linstow (1905: 534) places the form in the genus Strongyloides.

In June, 1904, I collected and preserved in formalin a large

number of specimens of the nematode in question from the large

intestine of a horse examined, post mortem, near Livingston, Mon-

tana, and have since collected it in the District of Columbia.

On the basis of this material I have found it possible to describe

this little nematode somewhat more fully than has been done hereto-

ee ee ee eee ee ee = —

ee

PROBSTMAYRIA VIVIPARA 35

fore, and as it presents rather marked differences from the types of

established nematode genera, I have ventured to propose for the

species vivipara a new genus, Probstmayria.

Genus PropstMayRIA Ransom, 1907

Generic Diagnosis.—Small spindle-shaped forms truncated ante-

riorly and with a long, gradually attenuated, acutely pointed tail.

Cuticula thin, transparent, and free from spines or setae. Mouth

with six lips. A small lateral organ of unknown function in rela-

tion with each lateral lip and the anterior portion of the body.

Pharynx elongated, cylindrical. Esophagus with two portions, an-

terior portion long and slender, posterior portion shorter, pyriform,

with a bulb containing a denticular apparatus. Anus about two-

thirds the length of the body from the anterior end. Male with two

short, nearly equal spicules, without bursa, and with a number of

pairs of small postanal papillae. Vulva of female at about the

middle of the body. Eggs few, hatching in the uterus, and devel-

oping into embryos which at birth exactly resemble the parent except

that the sexual organs are undeveloped.

Type species: Probstmayria vivipara (Probstmayr, 1865) Ran-

som, 1907.

Probstmayria vivipara (Pobstmayr, 1865) Ransom, 1907

SYNONYMY:

Oxyuris vivipara Probstmayr, 1865: 178-180. Ziirn, 1882: 252,

253. Perroncito, 1882: 337. Idem, 1886: 300-301 (under the

name Ossiuride vivipara). Idem, (?) 1902: 424-425. Neumann,

1888 : 370, 375. Idem, 1892 (French ed.) : 393, 399. Idem, 1892

(English ed.) : 403, 410. Fiorentini, 1890: 21-25, Pl. Railliet,

1887: 48 (as synonym of Rhabdonema vivipara). Idem, 1893:

549 (as synonym of Anguillula vivipara). Schneidemiihl, 1896:

324.

Rhabdonema vwipara (Probstmayr) Railliet, 1887: 48.

Anguillula vivipara (Probstmayr) Railliet, 1893: 549. Vaullegeard,

1901: 136. Jerke, 1903: 113-127, pl. 1, figs. 1-9.

[?] Oxyuris sp., Looss, 1902: 119.

Strongyloides viviparus (Probstmayr) Linstow, 1905: 534.

Probstmayria vivipara (Probstmayr) Ransom, 1907: pl. 11, figs. 1-8.

Specific Diagnosis.—Small, transparent, 2.5 to 3 mm. long when

mature, spindle-shaped, gradually diminishing in diameter towards

36 B. H. RANSOM

both ends, truncated anteriorly, ending in a very acute point poste-

riorly. Mouth surrounded by a very transparent cuticular rim or

collar, consisting apparently of six bulbous lips, two dorsal, two

ventral, and one on each side [according to Jerke (1902) there are

but three lips]. A slender filament extends through each ventral

and dorsal lip from the base to the anterior surface upon which is a

very small papilla in relation with the distal end’of the filament. A

similar filament in each lateral lip(?). In relation with and partly

: mbedded in the outer surface of the base of each lateral lip the bul-

bous cuticular expansion of an elongated organ, the remainder of

which is located behind the mouth collar beneath the cuticula.

Diameter of mouth collar 18» in immature individuals to 224 in

males and parturient females, height 5 to 7 p. Pharynx elongated,

cylindrical, lined with thick yellowish cuticula; 40» long in imma-

ture individuals and in the male, 40 to 50 long in parturient

females, and 9 to 13» in diameter, including an outer muscular

layer ; diameter of the tube formed by the chitinous lining, 4 to 6p;

chitinous portion of the anterior one-sixth of pharynx roughened,

irregularly thickened, slightly bulbous, and lacking the muscular

layer, with a number of irregular, tooth-like projections extending

outward into the substance of the mouth collar; posterior five-sixths

of chitinous lining of pharynx transversely striated with fine regular

corrugations. Esophagus consisting of two portions, a long, slender

anterior portion, and a short pyriform portion with a cylindrical

stem and with a denticular apparatus in the bulb; length of anterior

portion 240 to 255m in immature individuals, 295 to 320p in par-

turient females, and 300 in the male; diameter of anterior portion

near its middle 18, in immature individuals and in the male, and

18 to 22, in parturient females; posterior portion 80 to 88h long

in immature individuals, 105 to 115 in parturient females, and

105 in the male; diameter of the esophageal bulb 36 to 4ou in

immature individuals, 50 » in the male, and 45 to 55 » in parturient

females. Intestine, at its beginning, usually very wide, with a large

lumen; more slender and nearly uniform in diameter, with a narrow

lumen, throughout the remainder of its almost straight course to

the anus; diameter of intestine at its beginning 64 to 76m and near

the middle 30 to 54m in immature individuals, in the male 95 p

at its beginning and 65, near the middle, in parturient females

100 to 115 at its beginning and 62 to 72 near the middle; in the

PROBSTMAYRIA VIVIPARA 37

female there is a short, narrow rectum surrounded by 3 or 4 large

gland cells. Anus 1.35 to 1.4 mm. from the anterior end of body

in immature individuals, 1.75 mm. in the male, and 1.8 to 2 mm.

in parturient females. A nerve ring surrounds the anterior portion

of the esophagus near its middle at a distance from the anterior end

of the body of 168 to 185, in immature individuals, 200 in the

male, and 200 to 215, in parturient females.. Ventral of the

esophageal bulb a small vesicle opening to the exterior through the

excretory pore which is situated on the ventral surface of the body

335 to 3600p from the anterior end in immature individuals, 425 p

in the male, and 415 to 440 in parturient females.

Male.—Length 2.7 mm. Width at level of nerve ring 60, at

excretory pore 80p, almost uniform (95,) from a short distance

posteriad from the excretory pore nearly to the anus, in the region

of the latter becoming reduced to 75, and gradually decreasing

toward the tip of the acutely pointed tail. A single testicle extend-

ing anteriad as far as the esophagus, posteriorly its deferent por-

tion uniting with the end of the intestine on its ventral surface to

form the cloaca opening at the anus. Spicules two, similar, slightly

curved and pointed, nearly equal in size,67 and 58 p long, respectively,

and 6.7 in diameter. On the ventral surface of the tail six pairs

of small papillae, the successive pairs being from 20 to 25 » apart,

and the last or anterior pair 80, posterior of the anus.

Female.—Immature individuals 2 to 2.2 mm. long, with sexual

organs not yet apparent, measuring in diameter 45 to 55, at the

level of the nerve ring, 58 to 67 at the excretory pore, 64 to 76h

at a point about half way between the excretory pore and anus,

and 50 to 55 at the anus. Parturient females 2.7 to 2.9 mm. long,

measuring in diameter 63 to 72, at the level of the nerve ring, 88

to 100,» at the excretory pore, 100 to 115, at a point about half

way between the excretory pore and anus, 88 to 112» at the vulva,

and 72 to 82 at the anus. Vulva in parturient females located

about half way between the two extremities of the body, 7. e., 1.35

to 1.38 mm. posteriad from the anterior end. Vagina very short.

Uterus bicornate, one horn extending anteriad, the other posteriad

from the point of junction with the vagina, size and extent depend-

ing upon stage of development of the embryos contained therein.

Ovaries two, each consisting of a short coiled tubule, continuous

with the distal end of each horn of the uterus. Eggs elongate,

38 B. H. RANSOM

oval, with thin shell; size of eggs in uterus 58 by 40p to 100 by

75, according to stage of development. Only two to four eggs

present in the uterus at the same time.

Life History—Reproduction sexual. The eggs hatch in the

uterus and the embryos develop to a length of 1.8 mm. or more

before they are born. At the time of birth the latter exactly

resemble the parent with the exception that the sexual organs are

not yet developed. The embryos apparently may develop directly

into adults without leaving the intestine of the horse. Life history

outside the host unknown.

Hosts.——Horse (Equus caballus) ; ass (Equus asinus).

‘Habitat.—Large intestine.

Geographical Distribution—Germany (Probstmayr, Jerke), Italy

(Perroncito, Fiorentini), ?>Egypt (Looss), Montana, District of Co-

lumbia (Ransom).

BIBLIOGRAPHY

FIORENTINI, ANGELO.

1890. Sullossiuride vivipara (Oxyuris vivipara Probstmayr) cenni

descrittivi. Boll. scient., Pavia, XII (1): 21-25, 1 pl. 3 figs. Marzo.

JerKe, H. W. M.

1902. Eine parasitische Anguillula des Pferdes. Arch. f. wissensch. u.

prakt. Thierh., Berl, XXIX (1-2): 113-127, pl. 1, figs. 1-0.

Linstow, OTTo.

1905. Strongyloides fiilleborni n. sp. Centralbl. f. Bakteriol., Parasitenk.

fetc.], Jena, 1. Abt., XX XVIII (5) : 532-534, 1 pl. figs. 1-6. 15 April.

Looss, ARTHUR.

1902. The Sclerostomidae of horses and donkeys in Egypt. Rec. Egypt.

Govt. School Med., Cairo: 25-139, 13 pls., figs. 1-172.

NEUMANN, L. G.

1888. Traité des maladies parasitaires non microbiennes des animaux

domestiques. Pp. xv + 673, 306 text-figs. 8°. Paris.

1892. Traité des maladies parasitaires non microbiennes des animaux

domestiques. 2. Ed. Pp. xvi+ 767, 363 text-figs. 8°. Paris.

1892. A treatise on the Parasites and Parasitic Diseases of the domesti-

cated Animals. Pp. xxiii+ 800, 364 text-figs. 8°. London.

PERRONCITO, EDUARDO.

1882. I parassiti dell’uomo e degli animali utili. Delle pit comuni

malattie da essi prodotte profilassi e cura relativa. Pp. xii 506, 233

text-figs. 8°. Milano.

1886. Trattato teorico-pratico sulle malattie pi comuni degli animali

domestici dal punto di vista agricolo, commerciale ed igienico metodi

sui migliori metodi di disinfezione dei vagoni. Pp. xxiv + 434, 220

text-figs., I pl. figs. 1-4. 8°. Torino.

PROBSTMAYRIA VIVIPARA 39

(?)1902. I parassiti dell’uomo e degli animali utili e le pitt comuni

malattie da essi prodotte. Profilassi e cura relativa. Pp. xv-+632, 276

text-figs., 4 pls. 8°. Milano.

PRoBSTMAYR, W.

1865. Oxyuris vivipara. Wehnschr. f. Thierh. u. Viehzucht, Augsburg,

IX (23): 178-180. 8 Juni.

RAILLIET, A.

1887. [Note to Lutz, 1887, pp. 47-48.] Rec. de méd. vét., Par., LXIV

fais we 41, Cl) 345, y eoulan

1893. Traité de zoologie médicale et agricole. 2. fid., fasc. 1. Pp. 736,

494 text-figs. 8°. Paris.

Ransom, B. H.

1907. Probstmayria vivipara (Probstmayr, 1865) Ransom, 1906. A

nematode of horses heretofore unreported from the United States.

Trans. Am. Microsc. Soc., XX VII: 33-40, pl. 1.

SCHNEIDEMUHL, GEORG.

1896. Lehrbuch der Vergleichenden Pathologie und Therapie des

Menschen und der Hausthiere fiir Thierarzte, Arzte und Studirende.

2. Lief, Vergiftungen: Die durch thierischen Parasiten hervorge-

rufenen Krankheiten des Menschen und der Thiere. Die Konsti-

tutionskrankheiten. Die Hautkrankheiten. Pp. 209-448. 8°. Leipzig.

VAULLEGEARD, A.

1901. Etude expérimentale et critique sur l’action des helminthes. I.

Cestodes et nématodes. Bull. Soc. Linn. de Norm., Caen, 5. s., IV:

84-142.

Zurn, F. A.

1882. Die tierischen Parasiten auf und in dem Korper unserer Haus-

siugetiere sowie die durch erstere veranlassten Krankheiten, deren

Behandlung und Verhiitung. Pp. xvi+316, 4 pls. 8°. Weimar.

40 B. H. RANSOM

EXPLANATION OF PLATE

Plate II

Fig. 1. Lateral view of male. X 54.

Fig. 2. Lateral view of posterior end of male, showing the terminal por-

tions of the intestine and vas deferens, the cloaca and its opening, the spicules,

and the post-anal papillae. >< 190.

Fig. 3. Dorsal view of anterior end of body, showing the dorsal lips

with their papillae. > 400.

Fig. 4. Optical horizontal section through anterior end of body show-

ing the lateral lips, the lateral organs, the pharynx, and beginning of eso-

phagus. > 400.

Fig. 5. Lateral view of anterior end of body showing the lateral lip,

lateral organ, dorsal and ventral lips, and papillae. xX 400.

Fig. 6. Lateral view of immature form in which the genital organs are

not yet developed. X 54.

Fig. 7. Lateral view of young female, showing an egg in one horn of

the uterus. XX 54.

Fig. 8. Lateral view of parturient female with uterus containing a well

developed embryo, a second less developed embryo, and two eggs. XX 54.

a

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POROCEPHALUS CONSTRICTUS IN A NATIVE

FILIPINO

By MAXIMILIAN HERZOG AND CHAS. B. HARE

(From the Biological Laboratory, Bureau of Science, Manila, P. I.)

Pentastomum. constrictum, a parasite belonging to the order Lin-

guatulidae (tongue-worms), has been found only a few times in

human beings and then only in negroes living in Africa or once in

a negro recently migrated from this continent. A specimen of this

tongue-worm has, however, now been found in a native Filipino

and has been identified as such by Dr. Ch. Wardell Stiles.

According to Braun? the Linguatulidae, or tongue-worms, which

were formerly believed to be helminthes, belong to the class Arach-

noidea (or spiders, to which belong also the mites). They have, in

consequence of their parasitic nature, been profoundly modified, so

that their body is long, worm-like, either flattened or cylindrical,

and more or less annulated. Head, thorax, and abdomen are not

differentiated. Anteriorly on the abdominal surface there is an

elliptical mouth, surrounded by a ring of chitin and leading into a

straight gut which terminates at the hind end. On both sides of

the mouth are two hooks which can be retracted into two depressions

or grooves. These hooks are generally looked upon as the rudi-

ments of two pairs of legs; however, it seems more rational to con-

sider them the remnants of the palpi labiales and palpi maxillares

(Stiles). The whole nervous system consists of a mouth-ring.

Sense organs do not exist, except papillae at the anterior end.

Organs of circulation or respiration do likewise not exist. The two

sexes are separate. In the smaller male the sexual orifice is sit-

uated on the ventral surface, but anteriorly; in the female, it is

near the anus. The Linguatulidae lay eggs which in an interme-

diary host give rise to four-legged larvae. There is a second larval

1Braun. Die Thierischen Parasiten des Menschen, p. 336. Wuerzburg,

1903.

42 MAXIMILIAN HERZOG AND CHAS. B. HARE

stage and during it the parasite gets into its final host, mammalian

or reptilian, in whose body the full grown linguatulids live as

parasites.

The best-known and most widespread species of the Linguatu-

lidae is Linguatula rhinaria (Pilger), known since 1802, and also

described under the names Polystoma taentoides (Rud.), Lingua-

tula taenioides (Lam.), and Pentastoma taentoides (Rud.). It has

been found in man and in the dog, wolf, fox, horse, and goat. In

man it has been encountered, generally in the larval stage, in the

liver, kidneys, spleen, and intestines (Zenker, Heschl, Virchow,

Wagner, Frerichs, Klebs, Zaeslin). The rare linguatulid which we

found here in Manila is described by Braun under the name of

Porocephalus constrictus (v. Siebold, 1852), and, according to this

author, is known only in the larval stage. It is milky white with

golden-yellow hooklets, has 23 rings, is 13 millimeters long and 2.2

millimeters wide, and with no thorns at the posterior margin of the

body-rings. Braun says that cases in man have been reported by

Pruner, Bilharz, Fenger, Aitken, Girard, and Chalmers; and that

Pruner also found the parasite in a giraffe.

Chalmers,’ it appears, has been the most recent (1899) contrib-

utor, recording a case of Pentastomum constrictum found in a negro,

by race a Mendi, from Sierra Leone, Africa, who was admitted, sick,

into the hospital of the Gold Coast Colony and who died after a few

days. At the necropsy large numbers of the parasite Pentastomum

constrictum were found in the lungs, liver, alimentary canal, and

peritoneal cavity. Chalmers gives the following review of the lit-

erature of the subject:

“ Pentastoma constrictum was first discovered in 1847 by Pruner

in Egypt in the livers of two negroes upon whom he made post-

mortem examinations. He apparently did not recognize the zoolog-

ical position of the parasite, but sent specimens to Bilharz and Von

Siebold, who published an account of it and gave it the name of |

Pentastomum constrictum in their text-book of zoology.

“The next recorded investigation is that given by Aitken of a

case under the care of Dr. Kearney. This observer records the

clinical appearance of a negro private of a West Indian regiment

who died in January, 1885, at Jamaica, but who had come from St.

Helena only eight months previously.

*Chalmers. A case of Pentastomum constrictum. The Lancet, I: 1715.

1899.

POROCEPHALUS CONSTRICTUS IN FILIPINO 43

“ After this there is apparently a long interval without any further

records until Girard published an account of the illness and death

from purulent meningitis of a Senegalese.”

According to Chalmers these were the only cases which, up to

1899, had been reported in human beings. From Braun and Chal-

mers’ statements it appears that all cases reported occurred either in

Africa or in negroes which came from Africa. Of his own case

Chalmers says that on admission to the hospital the patient was much

emaciated and very weak. He had a markedly large and protuber-

ant abdomen, the temperature was slightly above normal and he

suffered from a hacking cough with expectoration of a little mucous.

The physical signs which were present in the respiratory areas were

those of congestion of the lungs with impaired resonance on both

sides, and a mucoid expectoration in which no parasites were to be

seen. The spleen was very much increased in size, and the liver

was enlarged but not tender ; the feces were not examined. Nothing

was found in the blood. The patient soon after admission devel-

oped a temperature of 104° F., rapidly became worse, and died.

The parasitic affection was only recognized during the post-mortem

examination, and then a large number of parasites were found

in the lungs, liver, alimentary canal, and peritoneal cavity. A large

number of parasites were found in the small intestine, with the

exception of the duodenum. “ Nothing,” the author says, “is known

at present as to the life history of this parasite or as to the means

by which it enters the body. It appears to me that it enters by

some means into the alimentary canal and that through this it

proceeds to the liver and forms cysts, which appear to be dilated

biliary ducts, and into the peritoneum. It also seems as though the

lungs were affected through the trachea, as cysts therein appear

like dilated bronchi. The presence of the parasite in the human

body sets up inflammation of the lungs and peritoneum but it does

not appear to irritate the alimentary canal or the liver. The only

possible means of diagnosis would be the presence of the parasite

in the feces in cases of wasting and emaciation in negroes. The dis-.

ease appears to be confined to negroes and the described cases came

from the Island of St. Helena, the City of Kayes, on the river

Senegal, in the French Soudan of West Africa, and from the English

Colony of Sierra Leone. Pruner does not state where the two

negroes who died in Egypt came from.”

44 MAXIMILIAN HERZOG AND CHAS. B. HARE

The case in which a Pentastomum constrictum was found in

Manila was the following:

Juan Flores, a native Filipino, had been a convict in Bilibid

Prison. He had been a sufferer from bone tuberculosis and was

admitted to the prison hospital in April, 1905. The clinical diag-

nosis in his case (Dr. W. R. Moulden) was tubercular abscess of

the thigh. The patient died after being in the hospital for fifty-one

days.

Post-mortem examination (Hare) on June 24, 1905—anatomic

diagnosis: Tuberculosis of the right femur, interstitial nephritis with

pyelitis of the left kidney, parenchymatous nephritis of the right

kidney, cirrhosis of the liver; great oedema of the brain.

Only a piece of moderate size of the liver, which macroscopically

presented the picture of a typical atrophic cirrhosis was kept for

histologic examination. This piece was hardened in formalin and

when later small cubical pieces were cut out of it a cavity, the size

of a millet seed, was cut into. Here the parasite, which had been

partly destroyed by the cutting knife, was found. A further search

for any more cavities or parasites was negative. Sections from the

liver exhibited the histologic picture of an atrophic cirrhosis.

Neither another Pentastomum nor any Schistosomum japonicum}

were seen.

The parasite found in the liver was sent to Dr. Ch. Wardell

Stiles of Washington, to whom we are greatly indebted for its iden-

tification. Dr. Stiles in substance reported as follows:

“ The parasite from the liver of a native is a tongue-worm. There

are two species of this group known to be parasitic in man, namely,

Linguatula taenioides and Porocephalus constrictus. This latter

species is looked upon as the larval stage of Porocephalus armil-

latus. Although the specimen sent is quite injured, it could be defi-

nitely recognized as not being Linguatula taenioides, and, so far as.

I can determine, from an examination of the fragments, it is the

larval form, generally known as Porocephalus constrictus. The

complete synonymy of the species is as follows:

POROCEPHALUS ARMILLATUS Wyman.

Synonyms—adult forms:

Linguatula armillata Wyman.

*Since Woolley reported in Vol. I, No. 1 of the Philippine Journal of

Science the first case of Schistosomum japonicum in a native Filipino, Herzog

has encountered a second case, likewise in a native Filipino.

POROCEPHALUS CONSTRICTUS IN FILIPINO 45

Pentastomum polyzonum Harley; idem, Leuckart.

Larval forms:

Pentastomum diesingt Van Beneden.

Pentastomum euryzonum Diesing.

Pentastomum. leonis Wedl.

Pentastomum constrictum Von Siebold.

Pentastomum protelis Hoyle.

Linguatula constricta Pruner; idem, Bilharz.

The adult stage is reported from the African pythons, Python

sebai and P. bivittatus (P. molurus), also from Felis leo. The lar-

val form is reported for Probeles ciristatus, Cynocephalus maimon,

and Homo sapiens. Pruner also reports the larval stage for the

liver of the giraffe. So far as I am aware, this is the first case

found in the Philippines.”

Neumann,! who a few years ago investigated the subject of

linguatulid infection of mammals, comes to the following conclu-

sion: The larvae of Porocephalus in the bodies of mammals do

not show a tendency to leave their cysts, hence they probably finally

perish in them. However, an observation on a lionness shows that

they are not absolutely harmless and may cause death in an animal.

This conclusion is probably applicable to human Pentastomum

constrictum. infection, since in only one of the reported cases does

death appear to have been directly attributable to these parasites.

1Neumann. Sur les Porocephales du Chien, etc. Archives de Parasit.,

II: 356. 1899.

ae

24

THe STRUCTURE AND CLASSIFICATION” OF. THE

SIPHONALES

WITH A REARRANGEMENT OF THE PRINCIPAL NORTH

AMERICAN GENERA.*

By CHARLES E. BESSEY

WITH ONE PLATE

The Siphonales consist principally of those algae in which the

plant body is a coenocyte, or a row or mass of coenocytes, in some

cases forming a globoid, or a simple filament, in others a branched

filament, which may be free or united into a compound body. They

are generally attached below by a simple or branched, usually color-

less, rhizoid. The plants range in size from those which are barely

visible to the naked eye up to those which are many centimeters in

length. For the most part they are marine, inhabiting the shal-

lower bodies of salt water, especially in the warmer portions of the

earth.

Each coenocyte is made up of a parietal layer of protoplasm which

lines the hyaline cell wall and surrounds the large vacuole. In the

parietal layer are numerous small nuclei and small rounded or

angled chromatophores, which may be united into a reticulum.

Although many of these plants are generally without division walls,

they can and do form them when the necessity for doing so arises.

Thus when a filament has been injured so as to threaten the loss

of its protoplasm a cross partition is quickly formed some distance

* This paper is in continuation of a series begun some years ago, and pub-

lished in these Transactions. Systematically the groups taken up are as fol-

lows: Protophyta, Vol. XXV, pp. 89-104; Protococcoideae, Vol. XX VI, pp.

121-136; Conjugatae, Vol. XXIII, pp. 145-150; Desmidiaceae, Vol. XXII, pp.

89-06; Bacillariaceae, Vol. XXI, pp. 61-85; Phycomycetes, Vol. XXIV, pp.

27-54.

48 CHARLES E. BESSEY

back of the point of injury. So, too, in the reproductive processes,

whether asexual or sexual, partitions are formed in order to cut

off the reproductive from the vegetative coenocyte.

In their reproductive processes, as far as known, the Siphonales

range from the fusion of similar biciliated gametes (isogamy) to

the union of a motile sperm cell (spermatozoid) with a non-motile

egg (heterogamy). Zoospores, usually biciliated, are commonly

present, but they are still unknown in some families.

These plants have generally been regarded as constituting a well-

defined group, in which the coenocytic structure is characteristic,

but within the past few years it has been found that it is by no

means as sharply set off from the other green algae as was formerly

supposed. In fact, it now appears probable that the coenocytic

structure has been gradually attained by the formation of fewer

and fewer partitions in the succession of filamentous plants.

In this paper I have made an attempt to arrange the coenocytic

algae in accordance with the theory that they have been derived

from multicellular filamentous algae of the Ulotrichoid type, among

the Confervales, where the segments of the filament are true cells,

each having a single nucleus. Near to these must be placed the

Cladophoraceae, in which the segments of the filaments are more or

less elongated coenocytes, each of which contains from one to many

nuclei. It has been the usual practice of algologists to regard the

Cladophoraceae as falling within the Confervales, to which, indeed,

they are closely allied in their general structure, and still more in

their reproductive processes. Recently it has been suggested that

the Cladophoraceae should be united with two or three other fami-

lies (Sphaeropleaceae, Valoniaceae, etc.), into an intermediate group

(Siphonocladiales) between the strictly cellular and the completely

coenocytic orders. This is essentially what is done by Blackman

and Tansleyt who divide the order (“Series”) Siphonales into the .

Siphoneae (including the families Protosiphonaceae, Derbesiaceae,

Caulerpaceae, Codiaceae, and Verticillatae) and the Siphonoclada-

ceae (including the families Valoniaceae, Gomontiaceae, Clado-

phoraceae, and Sphaeropleaceae). Nor is this very different from

the treatment of these families by Oltmanns,? who follows the order

*A Revision of the Classification of the Green Algae, by F. F. Blackman

and A. G. Tansley, in New Phytologist, Vol: I; 1902.

* Morphologie und Biologie der Algen, by Friederich Oltmanns, Vol. I;

1904: Vol. II; 1905.

STRUCTURE AND CLASSIFICATION OF SIPHONALES 49

Ulotrichales with Siphonocladiales (including the families Clado-

phoraceae, Siphonocladiaceae, WValoniaceae, Dasycladaceae, and

Sphaeropleaceae), leading to Siphonales (including the families Co-

diaceae, Bryopsidaceae, Caulerpaceae, and Vaucheriaceae). In both

cases an intermediate group between the cellular filamentous algae

and those which are wholly coenocytic is recognized, and in this

group are placed Cladophoraceae and Sphaeropleaceae. |

The arrangement in the present paper agrees with that of Black-

man and Lansley, and that of Oltmanns in including Cladophoraceae

and Sphaeropleaceae in the Siphonales, and placing them near the

lower end of the series. I have not, however, found it possible to

retain the two sub-orders, Siphoneae and Siphonocladaceae, since

to do so would separate families that appear to me to be distinctly

related, while at the same time it would bring together families

that have only a general relationship, especially in following Olt-

manns’s system.

In a general way I regard the Cladophoraceae as derived from the

Ulotrichaceae. The Cladophoraceae have given rise to two prin-

cipal phyla, the one characterized by the retention of a distinctly

filamentous structure, while in the other the coenocyte undergoes

great differentiation into “root,” “stem,” and “leaves.” The first

(including Sphaeropleaceae, Phyllosiphonaceae, Codiaceae, and Vau-

cheriaceae) culminates vegetatively in the complex plant body of the

higher Codiaceae, and sexually in the heterogamous Vaucheriaceae.

The second phylum (including Valoniaceae, and its miniature,

Botrydiaceae ; Derbesiaceae, Bryopsidaceae, Caulerpaceae, and Dasy-

cladaceae) finds its highest expression in the verticillate-branched,

basally rooted coenocytes of the Dasycladaceae. These relations are

roughly indicated by the accompanying diagram (Plater) ente

we designate these phyla by their highest representatives they may

be referred to as (1) the Vaucheria series, and (2) the Acetabularia

series.

The evolution of Cladophoraceae from Ulotrichaceae has been

almost wholly confined to the modification of the vegetative part

of the plants, resulting in less segmentation of the filaments, and

the consequent formation of elongated coenocytes. Asexual repro-

duction (“propagation’’) and sexual reproduction (“ generation ”’)

are essentially unchanged. The family Valoniaceae is a transition

group (if indeed it may not be regarded as including several

50 CHARLES E. BESSEY

groups) in which there is much diversity of form, but one can make

out at least an increasing tendency to non-septation of the filaments.

In Derbesiaceae we have a beginning of that regularity of the

branching of the erect filament (stalk) which is emphasized in

Bryopsidaceae, reaching its culmination in the extreme regularity

of the whorled branches of Dasycladaceae. Our very fragmentary,

knowledge of the modes of reproduction makes it impossible to

speak with certainty as to the evolution of asexual and sexual.

organs, but it must be said that what we do know indicates that

the development of these plants has been almost wholly in the struc-

ture of the plant body. It need only be said that the true position

of Caulerpaceae is problematical. All that we can yet say is that

it constitutes a side line which had its origin above Valoniaceae,

and certainly much below Dasycladaceae. The little family Botry-

diaceae is retained with its doubtfully related genera. It appears to

be related to the Valoniaceae, and for the present is placed near that

family. It should probably be broken up and its genera distributed

elsewhere.

From Cladophoraceae to Sphaeropleaceae the development has

been almost wholly confined to the sexual reproductive process.

There has been such a differentiation of the gametes that instead

of the free-swimming isogametes of Cladophoraceae we have small

free-swimming biciliated spermatozoids, and large, rounded non-

ciliated eggs, the latter not escaping from the pluriovulate gametan-

gium. The filament of coenocytes has undergone little change from

the type which is common in Cladophoraceae. In passing from —

Cladophoraceae to Vaucheriaceae the plant body has become almost i

completely non-septate, and at the same time there has been an im- —

provement over the heterogamy of Sphaeropleaceae in the special-|

‘ation of the antherid, and the reduction in the number of eggs in ©

the gametangium from several to one. The multiciliated zoospore :

of Vaucheria may be regarded as derived from the zoospores of Cla- ©

dophoraceae by a fusion of all of the biciliated zoospores of a Z00- i

sporangium into a compound, ovoid body, in which the component y

zoospores retain their individual cilia. The Codiaceae have devel- —

oped the plant body rather than the reproductive organs, and here, a

after the filaments’ became wholly non-septate, they were aggre i

gated into a plant body. The evolution of a more and more com-—

plex structure of the plant body is the characteristic feature in the

STRUCTURE AND CLASSIFICATION OF SIPHONALES np

Codiaceae. They are related to the other families in this phylum

in the tubular structure of their filaments, which are in others free

and separate, while here they are interwoven into definite forms.

From our meager knowledge of the zoospores and gametes it appears

that they are of the type of those occurring in the Cladophoraceae,

beyond which they have advanced only in reaching a differentiation

in the size, but not the form, of the gametes in some genera. Not

enough is known of the Phyllosiphonaceae to enable us to assign

them with certainty to a place in the system. They have generally

been regarded as related to the Vaucheriaceae, and on the supposi-

tion that they may have suffered degradation from a Vaucheria-like

type they are here placed near that group.

Order SIPHONALES

Plants coenocytic, filamentous, or saccate, often much branched,

and usually basally attached by rhizoids, from septate (consisting

of rows of coenocytes) to non-septate, the filaments single or aggre-

gated into a plant body of definite form: chromatophores discoid

or reticulated, parietal; propagation by: (1) The internal division

_ of the protoplasm of a part (sporangium), or of the whole plant

into spores,—in water into zoospores,—in the air into walled spores ;

or by (2) the contraction of definite masses of protoplasm into

agamic resting-spores (aplanospores or chlamydospores) ; genera-

tion by the union of (1) ciliated isogametes, (2) ciliated hetero-

gametes, or (3) spermatozoids with non-ciliated gynogametes (eggs),

or of (4) antherid nuclei (non-ciliated) with eggs, in all cases pro-

ducing zygotes.—Typically fresh-water and marine algae (holo-

phytes), from which many filamentous fungi (hysterophytes) have

been derived. (The latter are described in Volume xxtv of these

Transactions, pp. 27 to 54.)

There are eighteen pretty well marked families, of which eleven

are holophytic (algae), and seven hysterophytic (fungi). The algae

only are characterized in the following key.

KEY To THE FAMILIES.

A. Plants filamentous, septate, consisting of rows of coenocytes,

I. Filaments simple or branched, basally attached; isogamic,

Cladophoraceae.

II. Filaments simple, unattached; heterogamic, Sphaeropleaceae.

B. Plants filamentous, irregularly branched, non-septate,

I. Endophytic; no gametes known, Phyllosiphonaceae.

52 CHARLES E, BESSEY

I].Aquatic or terrestrial,

a. Filaments more or less compacted into a large plant body; isogamic,

Codiaceae.

b. Filaments single, free; heterogamic, V aucheriaceae.

C. Plants globular, minute, non-septate, terrestrial or aquatic, basally attached

Botrydiaceae.

by rhizoids,

D. Plants non-septate when young, usually becoming septate and compound when

mature; marine; zoospores biciliated, V aloniaceae.

E. Plants non-septate, regularly branched ; marine,

I. Filaments sparingly dichotomous, erect, with basal rhizoids; zoospores

multiciliated, Derbesiaceae.

II. Filaments pinnately branched, erect, with basal rhizoids; gametes bicili-

ated, Bryopsidaceae.

III. Filaments large, branched, creeping, with lateral rhizoids, and bearing

erect “ leaves,” Caulerpaceae.

IV. Filaments erect, regularly branched in whorls, with basal rhizoids,

Dasycladaceae.

FAMILY CLADOPHORACEAE

Plants filamentous, septate, simple or branched, mostly basally

attached by rhizoids; coenocytes with two to many nuclei, chroma-

tophores parietal, many, or united into a single reticulum; propa-

gation by (1) bi- or quadri-ciliated zoospores, which are produced

in undifferentiated segments, and (2) thick-walled aplanospores

developing from single segments; generation by the union of bi-

ciliated isogametes.

Key To THE GENERA."

A. Filaments unbranched,

I. Rhizoids at base of filaments,

a. Zoospores with four cilia, Urospora.

b. Zoospores with two cilia, Chaetomorpha.

II. Rhizoids lateral on the filaments, Rhizoclonium.

B. Filaments branched,

I. With quadriciliated zoospores, and biciliated gametes, Cladophora.

II. Only large, thick-walled aplanospores known, Pithophora.

III. Zoospores biciliated; plants minute, parasitic in shells, Gomontia. ©

1. Urospora Areschoug. Plant consisting of an unbranched,

basally attached filament, each segment of which is short and con-

1In the systematic treatment of the genera of Siphonales in this paper

while my plan has been to include only those known to have representatives

in America, I have not hesitated to include some foreign genera where they

help to a better understanding of the family, nor have I attempted to include

every genus, especially where they were either not common or had little

significance in the system.

STRUCTURE AND CLASSIFICATION OF SIPHONALES 53

tains several nuclei; zoospores ovoid, pointed, with four lateral

angles, and four sub-terminal cilia; zygote spherical—Species one,

in brackish or salt waters (New England). Filaments 1 to 8 centi-

meters long, and 10 to 70 (usually 20 to 40») broad.

2. Chaetomorpha Kuetzing. Plant an unbranched, basally at-

tached, thick-walled, filament, each segment short and containing

several nuclei; zoospores ovoid, pointed, not angled, and with two

sub-terminal cilia; gametes unknown.—Species many, mostly grow-

ing in brackish and salt waters, a few in fresh waters. Filaments

tufted or massed, coarse (40 to 500 broad) and rigid.

3. Rhizoclonium Kuetzing. Plant an unbranched, creeping, often

thick-walled, laterally attached filament, each segment short and

containing two to four nuclei; zoospores and gametes unknown.—

Species many, mostly growing in brackish and salt waters, a few in

fresh waters. Filaments entangled, 10 to 100» broad.

4. Cladophora Kuetzing. Plant usually a branched, basally at-

tached, thick-walled filament, each segment long and containing

many nuclei; zoospores ovoid, pointed and with four sub-terminal

cilia ; isogametes with two cilia——Species very many (200 to 300),

common in fresh, brackish, and salt waters. Filaments usually

coarse (25 to 30, to 400 to 500 broad), and forming large, float-

ing, entangled masses (“ water flannel ”) in strong currents of water.

5. Pithophora Wittrock. Plant a branched, thick-walled, basally

attached filament, each segment very long and containing many

nuclei; zoospores and gametes unknown; large thick-walled, ovoid

or barrel-shaped aplanospores occur, intercalated or terminally on

the filaments.—Species few, common in fresh waters. Filaments

coarse (50 to 175 » broad), the aplanospores larger and swollen.

6. Gomontia Bornet and Flahault. Plant a minute, branched fila-

ment, growing in the tissues of marine shells which they penetrate

by their rhizoids ; nuclei I to 5 in each segment ; zoospores pyriform,

with two cilia; gametes unknown.—Species one (New England),

inhabiting the tissues of living and dead molluscous marine shells,

to which they impart a greenish color.

FAMILY SPHAEROPLEACEAE

Plants filamentous, septate, simple, unattached (7. e., free-float-

ing), and without rhizoids; coenocytes much elongated, and con-

taining many small nuclei, chromatophores parietal, very numerous,

54 CHARLES E. BESSEY

disposed in rings at frequent intervals ; propagation unknown; gen-

eration by the union of biciliated spermatozoids with eggs which

remain within the gametangium and become zygotes.

There is but one genus.

1. Sphaeroplea Agardh. Plant an unbranched, unattached fila-

ment, each segment long and containing many nuclei; zoospores

unknown; spermatozoids narrowly ovoid, pointed, with two long

sub-terminal cilia, formed in great numbers in segments of the fila-

ment, from which they escape by lateral pores; eggs many in each

gametangium (oogone) ; zygotes red, germinating after a period of

rest, and then forming 2 to 8 rounded, biciliated zoospores, the latter

elongating into fusiform young filaments.—Species one, in fresh

waters. Filaments coarse (36 to 70 » wide), free-floating.

FAMILY PHYLLOSIPHONACEAE

Plants filamentous, non-septate, much-branched, living parasit-

ically in the leaf tissues of higher plants; nuclei many; chromato-

phores many, small, pale green; propagation by the formation of

numerous aplanospores in the branches; generation unknown.

There is but one genus.

1. Phyllosiphon Kuehn. With the characters of the family.—

Represented by but one species, which inhabits the parenchymatous

tissue of the leaves of Arisarum vulgare in southern Europe. It has

not yet been detected in North America.

FAMILY CODIACEAE

Plants consisting of branching, filamentous coenocytes which are

interwoven into an erect or decumbent general plant body of definite

form, which is rooted below ; chromatophores small, parietal; propa-

gation by biciliated zoospores formed in special branches; genera-

tion by the fusion of biciliated heterogametes. (Zoospores and

gametes are known in but few of the genera.) .

KEY TO THE GENERA.

A. Plant body cylindrical, spherical, or crustaceous,

I. Not stalked, cortical cells present, Codium.

II. Sometimes stalked, no cortical cells, Avrainvillea.

B. Plant body differentiated into stalk and crown or “leaf,”

I. Stalk cylindrical,

a. Crown of spreading, dichotomous, free filaments, Penicillus.

b. Crown of numerous small flat “leaves” of agglutinated filaments,

Rhipocephalus.

STRUCTURE AND CLASSIFICATION OF SIPHONALES 55

II. Stalk cylindrical or flattened, bearing a flat concentrically marked “ leaf ”

of agglutinated filaments, Udotea.

C. Plant body consisting of a branching series of wedge-shaped segments, usually

in one plane, Halimeda.

1. Codium Stackhouse. Plant body cylindrical and simple or

branched, or spherical, or crustaceous, attached basally by strong

rhizoids; inner filaments longitudinal, constituting the “ pith,’ and

giving rise to short, club-shaped branchlets which stand perpen-

dicularly to the surface and constitute the “cortex”; gametes pro-

duced in certain cortical branchlets, androgametes ovoid, smaller,

orange-colored, gynogametes rounder, larger, green.—Species many,

in tropical and temperate seas (South Florida and Pacific Coast).

Plant body from a few to many (5 to 30) centimeters long, spongy,

and dark green.

2. Avrainvillea Decaisne. Plant body an interwoven mass of fila-

ments, without differentiation into pith and cortex, and either with-

out a distinct stalk, or poorly differentiated into stalk and crown;

zoospores and gametes unknown.—Species few, in the warmer seas

(South Florida). Plant body several centimeters high.

3. Penicillus Lamarck. Plant body consisting of an upright,

cylindrical, basally attached, lime-encrusted stalk of interwoven fila-

ments, bearing a crown of spreading, dichotomous, free filaments ;

zoospores and gametes unknown.—Species about ten, in tropical

and sub-tropical seas (South Florida). Stalk from a few milli-

meters to several centimeters long; crown from one centimeter to

several centimeters in height and width.

4. Rhipocephalus Kuetzing. Plant body consisting of an upright,

cylindrical, basally attached, lime-encrusted stalk of interwoven fila-

ments, bearing an elongated crown of numerous small, flat “ leaves ”

of agglutinated filaments ; zoospores and gametes unknown.—Species

one, in tropical and sub-tropical seas (South Florida). Stalk 2 to 3

centimeters long; crown narrow, 6 to 8 centimeters high.

s. Udotea Lamouroux. Plant body consisting of an upright,

cylindrical, or flattened, basally attached stalk, bearing a flat, con-

centrically marked “leaf” of agglutinated filaments ; zoospores and

gametes unknown.—Species about ten, in tropical and temperate seas

(South Florida). Plants from a centimeter or so to 8 or Io centi-

meters in height.

6. Halimeda Lamouroux. Plant body consisting of a branching

series of wedge-shaped, lime-encrusted segments, composed of inter-

56 CHARLES E. BESSEY

woven filaments; segments usually in one plane, and the basal seg-

ment firmly attached by rhizoids; zoospores biciliated, produced in

rounded sporangia, marginal upon the segments.—Species many, in

tropical and temperate seas (South Florida). Plants from a few

to many (15 to 20 or more) centimeters in length.

FAMILY VAUCHERIACEAE

Plants filamentous, thin-walled, non-septate, and basally attached

by rhizoids; nuclei minute and very numerous; chromatophores

parietal, small, rounded, and very numerous; propagation by large,

oval, compound zoospores, produced singly in the ends of branches;

generation by the union of minute biciliated spermatozoids with

large eggs which remain within the lateral gynogametangia and

become thick-walled zygotes.

There is but one genus.

1. Vaucheria DeCandolle. Plant a bright green, branched fila-

ment, attached by a basal rhizoid; compound zoospores formed in

the end segments, cut off by a partition, and composed of very many

biciliated zoospores fused into an oval body, which escapes from the

segment by an apical opening, swimming for a time, then coming

to rest, withdrawing its cilia, covering itself with a cell wall and

elongating into a new filament; antherids lateral, forming many

minute, pointed spermatozoids, which have two lateral cilia; gyno-

gametangia (oogones) lateral, each forming one large egg; zygote

after a period of rest elongating into a new filament.—Species many,

common in fresh and brackish waters, or on wet ground. Filaments

coarse, soft-walled, usually felted (« green felt”), 5 to 20 centi-

meters long, and 50 to 200 p» broad.

FAMILY BoTRYDIACEAE!

Plants minute, globular or ovoid, non-septate, terrestrial or aquatic,

and basally attached by rhizoids; nuclei minute, many; chromato-:

phores parietal, minute and rounded, or united into a reticulum;

*The autonomy of this family may well be doubted. As said above

(page 50) it should probably be broken up and its genera distributed else-

where. Botrydium may eventually be placed in the Valoniaceae, near to

Halicystis. Whether Protosiphon is sufficiently distinct to be separated from

Botrydium can only be told after the latter has been as thoroughly studied as

the former. That Codiolum should be removed to Protococcaceae is most

likely. f

STRUCTURE AND CLASSIFICATION OF SIPHONALES 57

propagation by (1) zoospores, each with one or two cilia, (2) aplano-

spores which form in the rhizoids as well as in the plant above

ground; generation by the union of biciliated isogametes.

Kry TO THE GENERA,

A. Rhizoids freely branched, plants terrestrial, Botrydium.

B. Rhizoids simple,

I. Plants terrestrial, Protosiphon.

II. Plants aquatic, marine, Codiolum.

1. Botrydium Wallroth. Plant a minute, globular, green vesicle

on the surface of the ground, terminating below in a branched,

colorless rhizoid which penetrates the earth; chromatophores many,

minute; zoospores formed in the presence of sufficient moisture;

aplanospores formed when moisture is deficient; gametes not cer-

tainly known.—Species one or two, common on moist ground.

Globules 0.5 to I millimeter in diameter.

2. Protosiphon Klebs. Plant a minute, globular, green vesicle

on the surface of the ground, terminating below in a simple, color-

less rhizoid which penetrates the earth; chromatophores united into

a reticulum; aplanospores present; isogametes uniting to form

zygotes.—Species one. Globule about 0.2 millimeter in diameter.

This plant, which had been confused with Botrydium until sepa-

rated by Klebs in 1896, is now commonly regarded as quite distinct

from that genus, but if we maintain the family Botrydiaceae, I see

no need of transferring it to another family as is now usually done.

3. Codiolum A. Braun. Plant a minute, ovoid, green vesicle, ter-

minating below in a simple, solid, colorless rhizoid ; chromatophores

united into a reticulum; zoospores biciliated; gametes unknown.—

Species half a dozen, growing among other marine algae on stones,

piles, etc. Globule from very minute to a millimeter or two in

length. ,

This genus is very doubtfully related to the two other genera.

It appears to have closer affinities with the stalked Protococcaceae,

such as Characium, and probably should be placed there, as has been

done by Oltmanns.?

*Die Bedingungen der Fortpflanzung bei einigen Algen und Pilzen, by

Georg Klebs; 1806.

* Morphologie und Biologie der Algen, by Friederich Oltmanns, Vol. I;

1904: Vol. II; 1905.

58 CHARLES E. BESSEY

FAMILY VALONIACEAE

Plants filamentous and non-septate when young, basally attached

by rhizoids, usually becoming septate and branched, and often com-

pound when mature, the segments containing many nuclei; chro-

matophores parietal, small, numerous, rounded or angular; propa-

gation by (1) biciliated zoospores, and (2) aplanospores ; generation

unknown.—Marine plants.

“There is scarcely any family to which it is more difficult to

assign distinctive general characters, owing to the much varied

structure of the vegetative organs, and our ignorance of the repro-

ductive process in most of the genera. The thallus ranges in variety

from a single large cell [coenocyte] with rhizoids up to forms of

complex structure with stalk and frond’”’—(Murray’s Introduction

to the Study of Seaweeds).

KEY TO THE GENERA.

A. Plants septate and branching at maturity,

I. Septa at the bases of the branches,

a. But little and irregularly branched, Valonia.

b. Much and regularly branched, the branchlets uniting by rhizoids into

more or less definite shapes,

1. Branchlets united into stalked, leaf-like plants,

a. Branchlets forming an open network, Struvea.

b. Branchlets forming a solid structure, Anodyomene.

2. Branchlets united into leaf-like plants, which are sessile or pro-

cumbent, and attached by central rhizoids,

a. Segments (branchlets) alike, Microdictyon.

b. Segments (branchlets) unlike, long and short, Cystodictyon.

3. Branchlets uniting into irregular masses, Boodlea.

II. No septa at the bases of the branches,

a. Plant filamentous or columnar, Siphonocladus.

b. Plant a non-septated stalk, bearing a head of interwoven branches,

Chamaedoris.

B. Plants septate, unbranched, consisting of a layer of hexagonal cells united by

haptera, Dictyosphaeria.

C. Plants strictly non-septate,

I. Globoid, attached by a disk, Halicystis.

II. Cylindrical, much branched, : Apjohnia.

1. Valonia Ginnani. Plant irregularly tubular or vesicular, little

and irregularly branched, with a septum at the base of each branch;

zoospores biciliated, produced in ordinary cells; aplanospores pres-

ent.—Species many, marine, in the warmer seas (South Florida?).

Plants from a few millimeters to several centimeters in height.

STRUCTURE AND CLASSIFICATION OF SIPHONALES 59

2. Struvea Sonder. Plant consisting of a non-septate, erect

stalk, which bears a leaf-like structure composed of the regularly

pinnate branchlets which have united into an open network ; rhizoids

basal, well developed ; zoospores unknown.—Species few, marine, in

the warmer seas (South Florida?). Plants tufted, from a few mil-

limeters to one or two decimeters in height.

3. Anodyomene Lamouroux. Plant consisting of a short, erect,

compound stalk, which bears a leaf-like structure composed of re-

peatedly radiate branchlets which have united into a solid tissue;

zoospores not certainly known.—Species few, marine (South Flor-

ida). Plants a few centimeters high.

4. Microdictyon Decaisne. Plant a leaf-like, procumbent struc-

ture composed of irregularly placed similar branchlets, which have

united into an open network; rhizoids short, central; zoospores not

certainly known.—Species few, marine (Mediterranean Sea to Phil-

ippine Islands). Plants several to many (30) centimeters in diam-

eter, each forming a filmy, expanded network on the surface of rocks

or other algae.

5. Cystodictyon Gray. Plant a sessile or procumbent leaf-like

structure composed of irregularly placed, dissimilar (long and short)

branchlets, which have united into an open network; rhizoids short,

central; zoospores unknown.—Species one or two, marine_( Chinese

coast). Plants one to several centimeters in diameter. |

6. Boodlea Murray and DeToni. Plant an irregular spongy mass

composed of irregularly placed branchlets which have united into an

open network; rhizoids apparently wanting; zoospores unknown.—

Species one, marine (Japanese coast). Plants small.

7. Siphonocladus Schmitz. Plant at first non-septate, later abun-

dantly septate, erect or prostrate, sometimes saccate, often much-

branched; rhizoids basal, well developed; zoospores biciliated.—

Species few, marine (South Florida). Plants usually one to several

centimeters high.

8. Chamaedoris Montagne. Plant consisting of a non-septate,

erect, constricted and corrugated stalk, bearing a bushy-spreading

or cup-like head of interwoven branches; rhizoids at base of stalk;

zoospores unknown.—Species one, marine (South Florida). Plants

tufted, 3 to 5 centimeters high.

9. Dictyosphaeria Decaisne. Plant at first a hollow, cellular

globule with a basal rhizoid, at length bursting into an expanded

60 CHARLES E. BESSEY

layer of hexagonal cells united by haptera; zoospores unknown.—

Species few, marine (South Florida). Plants when mature irreg-

ular in outline and several centimeters in diameter.

10. Halicystis Areschoug. Plant non-septate, spheroidal, attached

by a stalk with a disk (rhizoid) below; zoospores unknown.—Spe-

cies one, marine (European). Plants 4 to 8 millimeters high.

11. Apjohma Harvey. Plant non-septate throughout, erect, cylin-

drical, repeatedly branched into a dendroid structure, with constric-

tions at the points of origin of the branches; rhizoids basal, well

developed; zoospores unknown.—Species one, marine (Australia).

Plants tufted, 8 to 15 centimeters high.

FAMILY DERBESIACEAE

Plants consisting of simple or sparingly dichotomous, erect, coeno-

cytic filaments, which are rooted below; chromatophores small, oval,

numerous; propagation by the production of zoospores in short,

rounded, lateral branches, each zoospore broadly ovoid, with a crown

of many cilia; generation unknown.

There is but one genus.

1. Derbesia Solier. Plant an erect filament, usually little

branched, bearing rounded lateral zoosporangia; zoospores with a

crown of cilia—-Species few, marine (South Florida). Plants

tufted, coarse, I to 5 centimeters high.

FAMILY BRYOPSIDACEAE

Plants consisting of erect, coenocytic stems, rooted below, and

regularly pinnate-branched above, the branches again sub-divided

into branchlets, the whole plant non-septate ; chromatophores small,

oval or elliptical, parietal; propagation unknown; generation by the

union of biciliated heterogametes which are produced in lateral

branchlets.

There is but one genus.

1. Bryopsis Lamouroux. Plant erect, very regularly pinnate-

branched, and attached below by well-developed rhizoids ; zoospores

unknown ; androgametes smaller, elongated, pointed, orange-colored,

biciliated; gynogametes larger, thicker, pointed, green, biciliated.—

Species many, marine (New York to Florida). Plants tufted or

single, from 2 to 15 centimeters high.

a 6 os

STRUCTURE AND CLASSIFICATION OF SIPHONALES 61

FAMILY CAULERPACEAE

Plants consisting of long, creeping, coenocytic, laterally rooted

stems which bear erect, simple, pinnate or variously lobed leaf-like

branches (“leaves”), the whole structure being non-septate, but

supplied with numerous “ cross-beams ” which prevent the collapsing

of the large coenocytes ; chromatophores small, parietal ; propagation

and generation wholly unknown.

There is but one genus.

1. Caulerpa Lamouroux. Plant a creeping rhizome-like stem

which is rooted at frequent intervals, and bears erect “ leaves”? which

may be simple or variously branched; neither zoospores nor gametes

are known, and it is supposed that new plants are produced by frag-

mentation—Species many (75 to 80), mostly in tropical and sub-

tropical seas (South Florida). Rhizomes from a few to many cen-

timeters long; “leaves” from small (a centimeter) to many centi-

meters high, and from simple, oblong, flat structures to those of

much complexity and beauty of pattern.

FAMILY DASYCLADACEAE

Plants consisting of erect, filamentous coenocytes, which are

basally attached by rhizoids, and bearing a succession of whorled

branches; chromatophores many, small, parietal; propagation un-

known; generation by the union of biciliated isogametes which

may form directly in gametangia, or aplanospores may. form and

give rise to gametes.

Key TO THE GENERA.

A. Stem mostly covered with whorls of persistent branches,

I. Whorls naked, free, not encrusted with lime, Botryophora.

II. Whorls united into a cortex, which is encrusted with lime,

a. Stem simple, unbranched, Neomeris.

b. Stem repeatedly dichotomous in one plane, Cymopolia.

B. Lower portion of stem naked, upper part bearing one or more whorls of

deciduous branches, Acetabularia.

1. Botryophora J. G. Agardh. Plant a simple, erect stem bear-

ing very many whorls of mostly persistent free branches which are

again branched dichotomously or trichotomously, not united into a

cortex, nor encrusted with lime; zoospores unknown; gametes not

yet observed, aplanospores present.—Species one, in sub-tropical

seas (South Florida). Plants tufted, 2 to 6 centimeters high.

62 3 CHARLES E. BESSEY

2. Neomeris Lamouroux. Plant a simple, erect stem, bearing

upon its whole length close whorls of dichotomous branches which

are united into a cortex, and encrusted with lime; zoospores and

gametes unknown; gametangia present.—Species few, in tropical

and sub-tropical seas (South Florida). Plants tufted, 2 to 3 centi-

meters high.

3. Cymopolia Lamouroux. Plant consisting of a main stem

which is repeatedly dichotomous in one plane, and covered for its

whole length with close whorls of dichotomous or trichotomous

branches which are united into a cortex and encrusted with lime;

zoospores and gametes unknown; gametangia present.—Species two,

in tropical and sub-tropical seas (South Florida). Plants tufted,

6 to 10 centimeters high.

4. Acetabularia Lamouroux. Plant consisting of a naked stem

which bears one or more whorls of deciduous branches on its upper

portion; fertile branches simple and agglutinated into a disk; zoo-

spores unknown; gametes biciliated, formed in aplanospores which

are produced in the disk branches.—Species few, in tropical and sub-

tropical seas (South Florida). Plants scattered or tufted, 5 to 12

centimeters high, bearing disks from 0.4 centimeter to 2 centimeters

in diameter, the whole plant encrusted with lime.

EXPLANATION OF PLATE

Plate III

CHART TO SHOW THE MUTUAL RELATIONSHIPS OF THE FAMILIES OF SIPHONALES.

The dotted line below marks the boundary between the Confervales and

Siphonales.

PLATE Ill

DASY CLADACEAE

CAULERPACEAE VAUCHERIACEAE

im

: CODIACEAE

a

Bee ee PHYLLOSIPHONAGEAE

DERIBES IACEAE

SPHAE ROPLEACEAE

VALONIACEAE

BOTRY Bec

CLADOPHORACEAE

SIPHONALES

—_— eee ee eee

CONFERVALES

ULOTRIC HACEAE

an ae eae a -

a

THE PHYLOGENY OF BACTERIA

By F. D. HEALD

WITH ONE PLATE

The hazy notions which seem to prevail in regard to the light that

modern physiology and bacteriological methods throw upon the

origin of the smallest of Nature’s creation, the bacteria, is the motive

which prompts this article. It is a generally conceded fact that

any system of classification should be as nearly as possible in har-

mony with the phylogeny. If this principle is followed it seems

quite certain that any scheme of classification which places the bac-

teria as a sub-class under the artificial group of the Fungi (Vines,

- 1902) is hardly the proper expression of their position in the plant

kingdom. It seems to the author equally absurd to place the bac-

teria and the Cyanophyceae in a heterogeneous group of poorly

defined genera—poorly defined, at least, from the bacteriological

standpoint (Cohn, 1875; Bessey, 1904). Neither should I agree

with the statement in regard to classification of the bacteria given

by Hueppe (Hueppe—Jordan, 1898). ‘“‘ Botanists are almost unani-

mous in accepting the essential features of the position taken by

Cohn, DeBarry, and Hueppe.”

The basis for the classification which places genera of bacteria

and blue-green algae side by side is the assumption that bacteria

are degraded blue-green algae that have lost their chlorophyll and

consequently their photosynthetic properties. The assumption is

based rather largely upon the similarity of reproduction by fission

(“fission algae,” “fission fungi”), upon the forms of the cells,

and upon the cell-grouping or the cell colonies. Neither these simi-

larities nor some others that may be mentioned appear to be of

sufficient weight to justify such an association. The only logical

course is to place the bacteria in an independent group parallel

with the Cyanophyceae and the various other groups of Thallophyta.

64 F. D. HEALD

The bacteria are called “fission fungi’’ even at the present time

by various authors. It is needless to discuss the fact that the simi-

larities are purely physiological, and that in the line of the mode

of nutrition. That physiological similarities alone are of little value

as indicators of genetic development is generally conceded. On the

basis of morphological characters the various fungi and the bac-

teria represent widely divergent forms. The rather ingenious theory

of Miller (Miiller, 1898) that bacteria originate from the spermatia

of various fungi, and that of Hallier (Hallier, 1895), which claims

their origin from the plastids of fungus cells, may be mentioned.

Notwithstanding the fact that both of these theories are supported

by extended laboratory investigations, they must be regarded as the

vagaries of distorted imaginations. In recent years some of the

true bacteria have been supposed to be simply stages in the growth

of a hyphomycete. The discovery of branched forms of the tubercle

bacterium, and some others has led to the creation of a new genus,

Mycobacterwuum (Chester, 1901; Lehmann-Neuman, 1896). It is

extremely doubtful if these branched forms represent normal indi-

viduals. They must rather be regarded as crippled and hypertrophied

forms of members of the bacillus group, that is, as involution

forms (Fischer, 1900). Species of Streptothrix which were for-

merly classed with the bacteria are the only forms which show a

similarity to the hyphomycetes and may very properly be excluded

from the bacteria.

A much closer relationship appears when we compare the bacteria

and that group of the infusoria known as the Flagellata. This is

especially prominent in the case of the Erythrobacteria ; for example,

Chromatium and Rhabdochromatium bear such striking similarities

to some of the Flagellata that it seems almost impossible to separate

them. The relationship is still further indicated by the fact that

bacteriopurpurin has been found to be identical with the pigment in

Euglena sanguinea, one of the recognized flagellates (Butschli, —

1883). But bacteria in general and the Flagellata appear to show

an equally close relationship. Some of the similarities and differ-

ences will serve to draw attention to this relationship.

First in the mode of reproduction by fission and the formation

of endospores we find a striking similarity. The former is of very

much less value than the latter, which is a pronounced morphological

character. All bacteria do not produce endospores (only some spe-

PHYLOGENY OF BACTERIA 65

cies of Bacillus, Bacterium, and Spirillum), neither do all flagellate

infusoria produce endogenous resting forms (Oicomonas, Pleuro-

monas, and Chromulina), but. that method of spore formation is

exemplified in both groups, although its details may vary somewhat.

Again, in the comparison of the organs of locomotion the simi-

larity is evident. The types of flagellation in the bacteria find a par-

allel in nearly every case among the Flagellata. The polar arrange-

ment of the flagella, as in Pseudomonas, Microspira, and Spurillum,

where we find one, two, or a tuft, is a common arrangement for

many flagellates. Thus, Oicomonas has one polar flagellum, Bodo,

Chilomonas, Cryptomonas and others have two, while Tetramitus

has four. The peritrichous type of arrangement of the flagella

characteristic for Bacillus is paralleled by Multicilia for the F lagel-

lata. A third similarity or agreement that is worthy of mention is

the character of the external membrane of the cell. The nitrog-

enous character of the cell wall or cell membrane in bacteria is

unique among plants. The so-called “cuticle” of the infusoria is

nitrogenous in character, but is not a definite membrane from which

the protoplasm can be separated by plasmolysis as in the case of the

bacterial cells; it is only the modified external layer of the cell

protoplasm.

The main differences that should be noted between the bacteria

and these low forms of animal life are in the cell contents. In all

except a very few of the flagellate infusoria that are either marine

Or parasitic, a contractile vacuole is a characteristic structure; no

such feature has ever been observed in any bacterial cells, although

non-contractile vacuoles may be noted in some of the larger forms.

A nucleus is a constant structural element in the F lagellata, while

such a structure is never present in a true bacterial cell.

In the matter of nutrition we find many striking parallels between

the bacteria and the flagellates. Very great emphasis should not

be laid on the similarities of nutrition, but they may at least be men-

tioned. The discovery of chemosynthetic flagellates is not beyond

the realm of probability and will only serve to intensify the fairly

well pronounced morphological relationships, as it seems that such

forms would be nearer the ancestral type, at least as far as their

nutrition is concerned.

Some of our present day botanists make the statement without

reserve that bacteria are degenerate and degraded blue-green algae,

66 , F. D. HEALD

absolutely excluding the possibility of their independent origin.

Before the discovery of the chemosynthetic type of carbohydrate

manufacture such an unqualified assumption would have been ex-

cusable, and even fairly probable, but in the light of these recent

advances in the nutrition of certain soil bacteria (Micrococcus

nitrosus, Pseudomonas europea, Bacterium nitrobacter), the inde-

pendent development of the bacteria, especially of the Haplobac-

teria, seems highly probable.

A fairly close analysis of some of the morphological and struc-

tural characteristics of the bacteria and the Cyanophyceae might

still leave some doubts as to their relationship, if chemosynthetic

food manufacture were unknown. A brief review of the differences

as well as the similiarities of the two groups brings out very sug-

gestive, but not conclusive, evidence. First, as regards the internal

structure and organization of the protoplasmic cell contents. The

most perfected cytological methods have failed to reveal the pres-

ence of a nucleus in any bacterial cell, notwithstanding the fact

that several investigators of prominence have made claims in that

direction (Butschli, 1896; Fischer, 1897), and until recently the

majority of evidence was in favor of a similar condition in the blue-

green algae. Some recent exhaustive work (Kohl, 1903; Olive, ©

1904), however, renders this supposed similarity more unlikely, since —

both authors claim to have discovered a distinct nuclear structure :

in many of the Cyanophyceae. This nucleus, if present (Fischer, ©

1905), is of a primitive type and may well be taken to represent |

one of the first steps in the evolution of the typical structure. The i

bacterial organism, occupying a lower rung in the ladder of life, ©

has not yet risen to the point where such a morphological differen- —

tiation has become necessary in the life of the cell. If the nucleate i

cell is the true morphological conception of the blue-green algae, ]

it appears difficult to understand how the adoption of a different |

mode of nutrition has caused the disappearance of the nucleus, as :

would be necessary if the bacteria are really degraded blue-green ©

algae. The above-mentioned conception of the bacterial cell as a

more primitive form, seems to the author to be much more probable. P

Parasitism or saprophytism does not lead in other cases to a dis- }

appearance of the nucleus, although it may lead to its diminution ©

in size. :

The mode of spore formation is one of the fallen strongholds of 4

PHYLOGENY OF BACTERIA 67

the advocates of the degeneration or degradation from the Cyano-

phyceae. The common production of arthrospores by various spe-

cies of the two groups was regarded as one of the strong links in

the chain of evidence. The pathway of science is strewn with the

wrecks of discarded hypotheses and erroneous observations, and

here is no exception. Although we find noted investigators (De-

Bary, 1884, 1887; Hueppe—Jordan, 1899) as champions of the

arthrospore theory for bacteria, the general consensus of opinion

among modern bacteriologists is that their observations were the

result of distorted imaginations or a misinterpretation of what they

actually saw. The fact that some of the Cyanophyceae form resting

spores from certain cells of a filament was quite probably the guid-

ing impulse in the discovery of the so-called “ arthrospores ” in bac-

teria. The only bacterium in which arthrospore formation is at all

probable is Streptothrix mesenteroides.1 Granting, then, that the

arthrospore is a delusion (Fischer, 1900), the similarity of spore

reproduction has disappeared, since none of the blue-green algae

form endospores. It would seem rather difficult to understand how

these poor degraded blue-green algae have either gained or retained

the power of producing endospores while their more specialized

and normal progenitors have either lost or never acquired the power.

This condition of spore formation is perfectly intelligible if we

assume an independent origin of the bacteria and the blue-green

algae from a common primitive ancestor. It is suggestive to note

in this connection that the formation of the so-called “ aplanospores ”

by some of the Chlorophyceae offers a certain morphological simi-

larity to the production of endospores by bacteria.

The common method of vegetative reproduction by fission in bac-

teria and Cyanophyceae is no argument for the origin of bacteria

from blue-green algae ancestors. It may be one of the links in the

chain of evidence, but it is rather frail. This common method of

propagation is no more an indication of relationship than it is of

a degradation from the Protococcoideae among the Chlorophyceae,

where a similar propagation prevails as the primitive type, in addi-

tion to analogous, if not homologous, cell forms.

A comparison of the two groups in regard to motility indicates a

rather close relationship between some of the Trichobacteria and

*The spore formation in Crenothrix and Cladothrix is not included in

this statement.

68 F. D. HEALD

certain of the blue-green algae. Species of Beggiatoa and Oscil-

latoria offer a striking similarity, not only from the morphological

standpoint, but from their peculiar oscillating and progressive move-

ments. In general the Leptothrix forms are quite similar to the

Oscillatoriaceae. On the other hand, when the Haplobacteria are

considered, we find motile and non-motile forms among coccus,

bacillus, and spirillum groups that are actively motile because of

the development of special organs of locomotion, the flagella; but

in the motile Cyanophyceae, no special organs are developed, only

the peculiar and unexplained oscillating or gliding movements being

exhibited. If the Haplobacteria are degenerate Cyanophyceae, it

is a peculiar case of degradation that has resulted in such a remark-

able degree of specialization in the production of monotrichous,

lophotrichous, and peritrichous forms. Such an idea is certainly

not in harmony with the much more evident cases of degenerate

forms.

In the character of the cell walls we find a similarity which is

rather largely due to similar physical properties, while from the

side of chemical composition a difference must be noted. The more

or less mucilaginous modification of the cell wall in the Cyano-

phyceae is a prominent character, and compared with the zoogloea

formation by many of the bacteria has been used as an important

feature in indicating relationship. It should be noted that many

of the Protococcoideae among the Chlorophyceae also show a muci-

laginous modification of their walls. If, instead of laying emphasis

on the similar physical properties of the modified wall, we consider

the chemical differences, a considerable divergence is indicated.

The normal wall in bacteria is unique among plants in being of

the nature of a proteid, while in the blue-green and other algae it

is a carbohydrate (cellulose). In the modified walls or the jelly

masses the same chemical differences prevail in probably the major-

ity of cases. It is true that in some cases it has been claimed that

bacteria form cellulose walls, and also that the jelly masses are of

a carbohydrate nature (Migula, 1897). Even if the statements are

reliable, the great bulk of something like one thousand species never

rise to that distinction. It may be that degeneration would result

in the loss of power to form a cellulose or even of any kind of a

carbohydrate wall, but it does not seem to me that it would operate

in that way. At least if this were true, we might reasonably expect

PHYLOGENY OF BACTERIA 69

to find more of these poor degenerates that had not yet been degraded

to the nitrogenous level.

The main stronghold of the advocates of the degradation of the

blue-green algae is apparently the similarity of external forms and

cell grouping in the supposedly related forms. It seems to me that

this apparent similarity has been allowed greater value as an indi-

cator of relationship than is really justifiable. However, before men-

tioning in some detail these similarities, it should be remembered

that almost analogous forms may be selected from the Chlorophy-

ceae. In the first place, it should be kept in mind that the coccus

or globular forms, and the rod or cylindrical forms, of the cells are

the most easily assumed and normal forms for free cells. Also the

simplest possible unions of cells are filaments, or linear aggregates,

cell-plates or superficial aggregates, and cell-packets or cell-masses.

The morphological similarities of the bacteria and the blue-green

algae have been dwelt upon more or less since the time of Cohn,

and there seem to be botanists at the present who consider these

similarities all-sufficient in determining the line of descent. Begin-

ning with the coccus forms we may compare Micrococcus and

Aphanocapsa, the cell division taking place in three planes, but

forming irregular masses. The diplococcus forms are quite closely

simulated by a form like Synechocystis aquatilis. Again, the tetrad

forms of Micrococcus and Placoma and Merismopedia agree in the

formation of groups of four cells, although distinct colonies are

formed by the algae, hollow irregular colonies by Placoma, and cell-

plates by Merismopedia. The colorless relatives never form distinct

colonies. Streptococcus forms and many of the Nostocaceae, for

example Anabaena, present a striking similarity. The morpholog-

ical similarities of Nostoc and Streptococcus mesenteroides are even

more pronounced. Although we find among the Cyanophyceae such

forms as Gloeocapsa, Gomphosphaeria, and Microcystis, in which

the cell division takes place in three planes with the formation of

colonies of cells, none that form definite cubical packets like Sarcina

have ever been recorded. It may be of interest to note in this con-

nection that such a form has recently been discovered by Shantz

in his plankton studies of Colorado lakes.t_ Those who regard the

bacteria as degraded blue-green algae will welcome this as the long-

* The form has not yet been published.

70 F. D. HEALD

looked-for Sarcina form and will place it as another link in the

chain of morphological evidence.

Considering next the bacillus forms of the bacteria, it may be

noted that Aphanothece resembles quite closely many of the zoogloea-

forming rod bacteria. Gloeothece may be compared with the cap-

sule-forming species of Bacillus and Bacterium.

Cells or cell-filaments showing the spiral form are found in the

two groups, although the differences are somewhat greater than

similarities of external form would indicate. It is quite suggestive

to compare the more or less curved species of Dactylococcopsts

with species of Spirosoma; also Spirulina and Arthrospira with

Spirillum and Spirochaeta. It is certainly a possibility that these

similarities indicate a definite genetic relationship, but the value of

such a comparison is somewhat lessened by the fact that quite strik-

ing parallels may be found in the more primitive Chlorophyceae.

If the Trichobacteria alone could be considered the degradation

hypothesis might still remain unshaken, for Beggiatoa and Oscil-

latoria, Cladothrix and Scytonema, Crenothrix and Tolypothrix

show a remarkable similarity. The corespondence is equally pro-

nounced between Phragmidiothrix and Chamaesiphon.

The differences between the Haplobacteria and some of the Proto-

coccoideae are but little more pronounced than the differences be-

tween the same forms and the blue-green algae, while similarities

of morphological characters are manifest although perhaps to a less

degree. If there were no blue-green algae, it would not be long

until some Cohnian systematist would have such forms as Tetraspora,

Palmella, Apiocystis, and Pleurococcus arrayed with the coccus

forms of the bacteria.1_ The relationship of the rod-bacteria (Bacil-

lus and Bacterium) to such forms as Dactylothece and Chlorococcum,

representing capsulated forms, and Stichococcus, a non-capsulated

form, would certainly not be overlooked, for the similarities of form

and cell-grouping are certainly well marked. It is also quite pos- ~

sible that he would see in the gracefully curved Raphidiwm the pos-

sibilities of a Spirosoma or a Spirillum. a

It seems to the writer that a consideration of the foregoing state-

ments must at least shake the foundations of the degradation hypoth-

1Tt seems quite probable that Bacterium viride, Bacillus virens, and Bac-

terium chlorinum, minute chlorophyll-bearing forms, are not bacteria but

primitive Protococcaceae.

PHYLOGENY OF BACTERIA 71

esis, if not to render it highly improbable. This hypothesis is the

logical outcome of the former physiological necessity of considering

the first and most primitive plant forms as chlorophyll-bearing

(Allen, 1895). The natural supposition was that the bacteria, sapro-

phytic and parasitic, had originated from these chlorophyll-bearing

photosynthetic ancestors. With the present physiological knowledge

(Pfeffer, 1897) such a supposition concerning the primitive forms

is no longer tenable, since there are bacteria that are capable of

utilizing carbon dioxide (CO,) as one of the crude substances in

food manufacture without the presence of chlorophyll, and that by

the process known as chemosynthesis. The first colorless forms

were not dependents but were capable of leading an independent

existence. In such primitive forms we have then the ancestors of

the present bacteria and the blue-green algae. Starting with this

chemosynthetic, non-motile, coccus form, the development of the

modern bacterial branch has arisen, on the one hand, while the

evolution of the Cyanophyceae has proceeded along another and

somewhat parallel line. The possibility of the relation of the Pro-

tococcoideae as well as that of the Flagellata and the Chytridaceae

to these primitive forms is by no means excluded.

It is interesting to note that in each of the suggested lines of

development there are indications that the chemosynthetic mode of

nutrition has been abandoned for the photosynthetic. There are

undoubted flagellate infusoria that contain chlorophyll (Phacus,

Euglena) ; the Erythrobacteria that apparently represent somewhat

separated forms have made photosynthesis possible by the produc-

tion of a purple pigment having chlorophyll-like properties, while

the blue-green algae exhibit the typical photosynthesis of higher

forms. The physiological and morphological relationship of the

groups in question may be expressed as follows:

Photosynthetic nutrition

Saprophytic nutrition

Parasitic nutrition

Chemosynthetic nutrition (?)

Chemosynthetic nutrition

Photosynthetic nutrition

Haplobacteria.4 Saprophytic nutrition

Parasitic nutrition

Symbiotic nutrition

Photosynthetic nutrition

Saprophytic nutrition (?)

Trichobacteria Saprophytic nutrition only.

Flagellata. ....

Primitive bacterium: coccus

form, non-motile, nutrition

chemosynthetic.

Cyanophyceae.

72 F. D. HEALD

The relationships of the Trichobacteria must be looked upon as

rather problematical. It is the opinion of the writer that they are

not directly related to the Haplobacteria. If there are any bacteria

that are degraded blue-green algae, they are certainly the ones,

although the possibility of a direct descent from the Haplobacteria

should not be absolutely excluded.

Taking into consideration the morphological characters (Migula’s

Classification—Migula, 1900), the phylogeny of the Haplobacteria

may be expressed according to the diagram in the accompanying

plate.

BIBLIOGRAPHY

ALLEN, GRANT.

1895. The Story of Plants. D. Appleton & Co.

BEsseEy, C. E.

1904. Classification of the Protophyta. Trans. Am. Mic. Soc, XXV:

89Q-104.

BUTSCHLI, O.

1883. Bronn’s Classen u. Ordnungen des Tierreichs, I (2). 1883-1887.

1896. Weitere Ausfuhrungen iiber den Bau der rafentanra und Bac-

terien. Leipzig, 1896.

CHESTER, F.

1901. Determinative Bacteriology. Macmillan Co.

Coun, F.

1875. Beitr. z. Biol. d. Pflanzen, I (3).

De Bary, A.

1884. Morphologie u. Biologie der Pilze, Mycetozoen, u. Bakterien.

1887. Lectures on Bacteria. Clarendon Press; 1887.

FIscHer, A.

1897. Untersuchungen iiber den Bau der Cyanophyceen und Bakterien.

Jena, 1897.

1900. Structure and Functions of Bacteria. Clarendon Press; 1900.

1905. Die Zellen der Cyanophyceen. Bot. Zeitung, LXIII: 51-120.

HALLIER, E.

1895. Pestkrankheiten der Kulturgewachse. Verlag von Erwin Nageli;

Stuttgart, 1895.

HUEPPE-JORDAN.

1899. The Principles of Bacteriology. Open Court Pub. Co.; 1809.

Kou. :

1903. Ueber die Organization und Physiologie der Cyanophyceenzelle und

die mitotische Teilung ihres Kerns. Gustav Fischer; Jena, 1903.

LEHMANN-NEUMAN.

1896. Bak. Diag., 1806.

Micuta, W.

1897. System der Bakterien, I. Gustav Fischer; Jena.

1900. System der Bakerien, II. Gustav Fischer; Jena.

PHYLOGENY OF BACTERIA 72

MULLER.

1898. SBacterien und Eumyceten. Fischer; Berlin, 1808.

Otive, E. W.

1904. Beihefte zum Botanischen Centralblatt, XVIII (1): 9-44.

PFEFFER, W.

1897. Pflanzenphysiologie, I. Wilhelm Engelmann; 1897.

VINES, S.

1902. Student’s Text-book of Botany. Macmillan Co.; 1902.

EXPLANATION OF PLATE

Plate IV |

A Diagram to show the Phylogeny of the Haplobacteria.

PLATE IV

SpiTochaeta

seem Microspira

Spirosoma

Bacillus

i Pseudomonas

OOF

'

'

'

A

\

1

)

}

’

i

Bacterium

GED]

Planococcus

Planosarcina

Micrococcus 20

Streptococcus 46%

Primitive Form

A BIOLOGICAL STUDY OF THE LAKES OF THE PIKE’S

PEAK REGION—PRELIMINARY REPORT

By H. L. SHANTZ

WITH THREE PLATES

The Pike’s Peak region lies in the east central part of Colorado,

about 38° 30” north latitude and 105° west longitude. Here, within

a few miles of each other, are two groups of lakes representing

quite different types, the alpine and those of the plains (pl. v).

The alpine lakes lie far up on the mountains at altitudes varying

from 3110 m. to 3625 m., with typical alpine surroundings. The

plains lakes lie on the western edge of the Great Plains at altitudes

varying from 1800 to 2000 m., and with conditions which are in no

wise alpine. They are lowland lakes. |

THe ALPINE LAKES

In the region south and east of Pike’s Peak there were a number

of natural bodies of water which were known as Lake Moraine,

Dead Lake, and the Seven Lakes (pls. v and vr). In addition to

these there have been constructed from time to time a number of

reservoirs. Lake Moraine has been changed to a reservoir and

within the last two years Seven Lakes have been much altered by

the construction of dams which have converted three of these lakes

into one large reservoir (pl. vit), and have destroyed two others. Of

these lakes the writer chose six as representative and in addition a

small pond near the top of Bald Mountain. These seven lakes, from

which collections were made, were Moraine, Dead, Mirror, Ribbon,

Isoetes, Michigan, and Bald Mountain. Several collections were

also made from Marsh Lake.

These lakes, with the exception of Bald Mountain, have been

described by Ward (1904). In addition to those studied by the

writer, Ward also included Lake of the Rocks and Lake Ramona—

mere ponds—which were destroyed in the construction of Reservoir

No. 4.

76 H. L. SHANTZ

The alpine lakes studied, together with the elevation, the area,

and the depth of each, are given in the list below:

Lake Elevation Area Depth

Moraine 3110 m. 39.65 ha. 8.5 m.

Isoetes 3285 m. 1.00 ha. 2) om.

Michigan 3285 m. I.I ha. Lig:

Marsh 3287 m. 0.5 ha. 0.5 m.

Ribbon 3290 m. BATA: 4.5 m.

Mirror 32902 m. a3: tes 8 m.

Dead 3350 m. Tera ia: Mads v9 &

Bald Mountain 3625 m. 3. ha. (of small pools) 0.5 m.

(Reservoir No. 5 3292 m. SOP. i ha: 18 m.)

(Reservoir No. 4 3290 m. SO wena: Sey Oy

Reservoir No. 5 was built on the site of Marsh, Ribbon, and Mirror

lakes. Although the dam was constructed in 1905 the water did

not rise high enough to include Mirror and Ribbon lakes until the

spring of 1906. Collections were made from the original lakes up

to that time. The collections made during 1906 were, however,

from the reservoir and on the sites of the original lakes. During

1905 collections were taken from reservoir No. 4, but during 1906

this reservoir was dry.

As pointed out by Ward (1904: 138) these lakes are higher

than any from which plankton studies have been reported, either

in this country or in Europe. The difference in altitude between

these lakes and the European lakes studied by Zschokke (1900: 1)

is very marked. The highest lake included in his lists was Fuorcla

de Flex, 3050 m., which is 60 m. lower than any of the alpine

lakes studied by the writer. On the other hand, it is 242 m. below

Mirror Lake and 576 m. below Bald Mountain Lake. Zschokke

(1900: 2, 40) includes all lakes above 1500 m. as alpine, but such

a limit is not to be applied in this portion of North America, as

already pointed out by Ward (1904: 133). Latitude as well as

general climatic condition should be taken into account. Perhaps

a far better method of comparing alpine lakes would be to take

the distance above or below timber line. This line, although not

absolutely uniform, is to a great degree an invariable altitude in

each section of the country. In a way, it is a measure of the

alpine condition and a lake lying above timber line, area and depth

being equal, will show more extreme mountain conditions than one

lying below, even though the altitude is less.

LAKES OF PIKE’S PEAK REGION 77

Timber line on Pike’s Peak is at an elevation of 3573 m. (11,720

ft., Hayden Survey). In the following table the distances above or

below timber line are given:

Lake Moraine................ 363 m. below timber line.

Isoetes Lake...............-.- 288 m. below timber line.

DATCOIO ATEN AKON Yi chi cracer oa 288 m. below timber line.

erste ake bos kc riuleaene 290 m. below timber line.

Ribbon Lake................. 293 m. below timber line.

Wirerori Leakey 3/04: Woretur cc ores 295 m. below timber line.

BGA Lakedsin swum eer crac 275 m. below timber line.

Bald, Mountain Uakew 4.02. 52 m. above timber line.

Many of the lakes of the Alps lie above timber line and while

they do not have as high an elevation the alpine conditions are much

more extreme than are those in the region under consideration. A

glance at this table will show only one lake, Bald Mountain, above

timber line and this lake is really only a series of pools.

Bald Mountain Lake is surrounded by an alpine meadow, a

thick turf of mat plants and grasses. In addition to this there are

gravel slides which, however, do not reach to the water’s edge.

There are also many large rock fragments lying in and about the

ponds. Mirror and Ribbon lakes were largely surrounded by moun-

tain meadow, but a coniferous forest covers Bald Mountain, which

rises rather abruptly above the site of Mirror Lake, and this forest

formerly extended along the edge of Ribbon Lake. This portion

of the forest has since been cut away and the site of Ribbon Lake

is now several hundred meters removed from forest growth.

Isoetes Lake has a forest lying on the south side, Marsh Lake was

surrounded by swamps, and Michigan Lake is bordered by swamp,

thicket, and forest. Dead Lake lies near a dense coniferous forest,

but is also bounded by thicket and meadow (Shantz, 1905: 256).

Lake Moraine is bounded by thicket, coniferous and aspen forest,

and, on the lower side, by a dam. Bald Mountain and Michigan

lakes receive practically full sunlight, as did also Marsh Lake.

The others were shaded during part of the morning. This was

true of Mirror Lake especially.

Shore and Bottom.—The shore of Mirror Lake was of clean

gravel, with the exception of a small portion, where a certain

amount of plant remains and ooze was carried in from a swampy

area. There was no growth of higher plants and no algae were visi-

ble in the littoral region. The bottom of the lake was of blue

78 H. L. SHANTZ

clay. Ribbon Lake had practically the same condition of shore and

bottom as had Mirror Lake. In Isoetes Lake the shore line is partly

swampy and partly gravel, while the bottom is covered to some

depth with ooze. Within the last two years the water has been

filled with silt. This is due to the fact that Marsh Lake had been

drained into this lake and also that a reservoir was being con-

structed just above it. The level has been lowered so as to expose

the whole shelf of the lake. Here we find a dense growth of

Sparganium, forming a zone around the lake and also an inner,

much interrupted, zone of Batrachium. .

Michigan Lake is completely overgrown with Sparganiwm and

contains, in addition, a great deal of moss. The bottom is covered

to some extent with organic remains, but the net also brings up clay

and gravel. Dead Lake has a rather clean gravel and clay bottom,

but in certain places there is a considerable amount of organic ooze.

Lake Moraine has a clean gravel bottom, but at the upper end the

growth of Carex has produced great masses of peat which float out

into the lake and which are removed from time to time by workmen.

Water Supply—The water supply of Dead Lake and Michigan

Lake is entirely from springs, as was also that of Mirror Lake, with

the addition of the run-off from a very small area. Since the sur-

rounding area is well covered with vegetation, this surface water

does not bring an appreciable amount of silt to these lakes. Ribbon

Lake was supplied directly from Mirror Lake, while Marsh and

Isoetes lakes received the overflow of Ribbon Lake. Lake Moraine

is fed by Ruxton and Beaver creeks, the water of the latter being

ditched and piped into the former. Bald Mountain Lake is fed

entirely by springs. All of these lakes have an outflow with the,

single exception of Dead Lake which is a closed lake.

Temperature.—The following table gives the temperature read-

ings in Centigrade units recorded during Ig04:

Lake May 20 June4 Junex7 July 12 July 29 Aug. 12 Aug. 26 Sept.17 Oct.3 ©

Bald Mountain DAO. i 3 au © 6.8°

Dead Tt A rO.oiulere! Mi OLR ES Of) sh SOe il 3.5 nek SD 9°

Mirror 11-48 6 8 16 T2040 uT 210 oat 3 1T.520).10.8

Ribbon 113.2 0105.5 9 10:2 20 EA i sO WaT II 12

Michigan 12-58 6.5 Sa nee T4604 :0.16 15 15.2) /150

Tsoetes 98 5 9 TO) walOie an TS 13.0)5rw 15

Moraine I.I -6 8 OALM IS 13 13.3

On May 20 all of the lakes with the exception of Isoetes Lake

|

{

; 4

LAKES OF PIKE'S PEAK REGION 79

were partly covered with ice. The highest temperatures obtainable

at this time were taken near the edge in the most open part of the

lakes.

The high temperature recorded for Isoetes Lake on May 20, 1904,

was due to the drainage of Marsh Lake into this lake at that time.

On June 4 a heavy snow storm lowered the temperature of all the

lakes, and there was still some ice remaining at portions of the

shore. On June 17 Bald Mountain Lake was covered with a thin

ice sheet. |

In respect to temperature Zschokke (1900: 20) divides alpine

lakes into three categories: First, lakes of considerable size and

depth; second, ponds or small, shallow lakes; and third, glacial

lakes—lakes fed directly from ice and snow. Although much

smaller than the lakes considered by Zschokke the following may be

placed in the first category: Mirror Lake (4.8°-13.6°) Moraine

Lake (6°-13.3°), and Ribbon Lake (3.2°-14.8°). Dead Lake

(4.7°-15.6° ), although very small, does not properly belong in either

of the other classes and may be included here. All of the other

lakes belong to the second category: Isoetes (5°-17.6°), Bald

Mountain (1°-12.2°), and Marsh lakes.

~The temperature of these ponds is surprisingly low when we con-

sider that there are here many conditions favorable for a higher

temperature of the water. Doubtless one reason for the low tem-

perature of these ponds is to be found in the low relative humidity

of the air, the rarity of the air, and other climatic conditions which

bring about very rapid evaporation and the resulting cooling of the

water surface. The air temperature is, however, never very high.

‘Air temperatures taken at the time the water temperature was read

range from 0.2° at Michigan Lake, June 4, 3.30 p. m., to 19.4°

at Michigan Lake, July 12, 12.30 p.m. With the exception of the

last reading the highest temperature recorded was 16°. Another

reason for the low temperature of these ponds is to be found in the

low temperature of the spring water which supplies them. <A very

large spring which empties directly into Michigan Lake was found

to have a temperature below 6° even in the warmest part of the

year.

Temperature readings taken during 1905 and 1906 are about the

same as those given for 1904 for corresponding periods of the year.

In none of these lakes was a thermocline found. The bottom

So H.'L: SHANTZ

temperature differed very little from that of the surface. The fol-

lowing readings were made July 29, 1904:

Lake Surf. Im. 2m. 3m. 4m. 5 m. 8m.

Mirror 13.6° 13 On het ay 13.0 VoNEg.6% N00 5.5, cael

Ribbon 14.8 14.6 14.5 13.8 13.8

Dead 13.6 13.4

Readings made on September 17 did not show any relative change

in temperature.

For convenience we may divide these lakes into a number of

types—closed lakes, open lakes, ponds and reservoirs. Dead Lake,

the only closed lake studied, has been described in some detail by

Ward (1904: 131-135, pls. XxIv, XxXv, XxIx) and Shantz (1905:

256). Of the open lakes there were two—Mirror Lake (Ward, 1904:

132-135, pls. XXIV, XXV, XXVI, XXVII, XxvIII) and Ribbon Lake

(Ward, 1904: 131-135, pls. XXIV, XXV, XXVI, xxviI). Mirror

Lake was the larger and had no inlet but was connected with

Ribbon Lake through its outlet, and Ribbon Lake in turn, through

Marsh Lake and Isoetes Lake, with Beaver Creek and thence

ultimately with the Arkansas River. Lake Moraine is a reser-

voir but does not differ essentially from the open lakes just dis-

cussed except that there is no overflow, the water issuing from a

pipe at the bottom of the lake. This allows the bottom water to

escape rather than the surface and would not remove from the lake

the true plankton. The remaining lakes, Michigan, Isoetes, and

Bald Mountain, and formerly Marsh as well, may well be called

ponds. They are small, shallow, partly filled with plant growth and

organic ooze, and reach a relatively high temperature.

If we compare these lakes with the general description of a

typical alpine lake as given by Zschokke (1900: 40), many points

will be found to agree. The water basins lie above 1500 m. and are

small and of varying depth. The bottom and shore show manifold -

local differences as do also the outward conditions. Of the flora,

mosses and algae are abundant in the smaller pools. The littoral

flora is almost entirely wanting in the larger lakes. The inflow is

of water poor in nourishment (varying from 0.6° to 10° except in

the case of the lakes which are supplied by streams). The outflow

is underground in the case of Dead Lake and also Lake Moraine

(an artificial condition in the latter case). The surface is practi-

cally undisturbed. The water temperatures are low even in the

LAKES OF PIKE’S PEAK REGION 81

middle of the summer, surface temperatures being but little higher

than the deeper strata of the lakes of the lowland. The summer

maximum and the winter minimum lie close together. The period

of ice cover is of long duration. The chemical composition of the

water is variable.

A comparatively few points do not agree, but this is to be ex-

pected, since the description is of a typical alpine lake and is not

intended to include all the conditions which might be found in moun-

tain lakes. The chief points of difference are these: There is no

immediate danger of the extermination of these lakes by drought,

rock slides, or avalanches. The Characaceae do not occur and, in

the greater number, mosses and algae are not present in sufficient

numbers to be noticeable. Under natural conditions the level is

fairly constant for all lakes under discussion. In only one case is

the outflow subterranean.

To a certain extent glacial conditions are present in these lakes.

The period during which the lakes are covered by ice sheets has not

been definitely determined. It is of comparatively long duration.

On May 20, 1904, Mirror and Ribbon lakes were still covered with

the ice sheet, except at the very edges, while Moraine and Michigan

were partly covered. At Dead Lake the wind had broken up the

ice sheet in the early part of the day. Even on June 4 a consid-

erable amount of ice still remained on the protected shores of the

lakes. No attempt was made to collect from Bald Mountain before

June 17, at which date there still remained a portion of the old ice

sheet and the lake had been frozen over anew on the previous night.

THe PLains LAKES

These lakes are without exception artificial and have been built

largely for purposes of irrigation, with the exception of three, which

are used for the storage of drinking water.

Without exception these lakes lie in the open where they receive

full sunlight and are not protected in any degree from the wind.

The bottoms were originally very uniform. As a rule they were

built in gravel areas, but clay or cement has been used to prevent

leakage.

Those which were taken as types are given in the following table:

82 H. L. SHANTZ

Reservoir Elevation Area Depth (original) -

Mesa No. 1 (Palmer’s) 2000 m. 1.06 ha. 7.92 .m.

Mesa No. 2 (Palmer’s) 2000 m. 2.04 ha. 7 Geet,

Mesa No. 3 (Palmer’s) 2000 m. 4.04 ha. 9.14 m.

Prospect (Lake) 1830 m. 20.94 ha. 0.14:

Portland 1820) Mine cpa 3,5 anal

Boulder 1829 m. fae has TSa0nar,

Colorado Springs No. 2 1890 m. 1.61 -“ha. 5.48 m.:

Colorado Springs No. 1 1886 m. Ora: 3.64' m.

Colorado Springs (on Mesa) 1900 m. 1.82* ha. 6.1* m.

Becker’s 1950 m. 1.07 na. 2.43* m.

Broadmoor 1950 m. Asa nae 2.43* m.

Jenk’s (pond) 1800 m. Ts sania TLOts im,

* Approximate.

The altitudes given are also only approximate. They are computed from

the U. S. topographical map.

Besides those given in the table above, collections were made from

two small reservoirs in Stratton Park, one at Pike View, and a

number of other small nameless reservoirs. The depths given are the

original depths and in the case of the older reservoirs, this figure

probably exceeds the present depth. In Prospect Lake, which origi-

nally had a depth of 9.14 m., the writer was unable to find a depth

greater than 5.5 m. It is probable that measurements of the other

reservoirs would have also shown that there had been some filling

in since the first construction.

The following table gives the age, in 1904, counting from the year

first filled, as well as the source of the water supply:

Reservoir Age in Years Source of Water Supply

Colorado Springs No. 1 28 Ruxton, Beaver, and Bear creeks,

through city water system.

Colorado Springs No. 2 TON RGAssabove!

Colorado Spgs. (on Mesa) rf Ruxton and Beaver creeks, through

city water system.

Prospect 13 As above.

Mesa No. 1 5 Camp Creek, through Glen Eyrie water

system.

Mesa No. 2 5 As above.

Mesa No. 3 I As above.

Broadmoor 17 Cheyenne Creek.

Portland 4 Bear Creek, through a ditch.

Becker’s 16 Sutherland Creek.

Boulder 26 Fountain Creek, through El Paso ditch. —

Jenk’s very old Small, unnamed stream. |

*First filled in 1904.

LAKES OF PIKE’S PEAK REGION 83

The following table gives the temperatures recorded during 1904:

‘ Date Ae Rhea Prospect Boulder Portland Sereee: ee

May 19 2. Wea

23 a LL.O:

27 13.6° TIO.

June 8 15.0° 14.8° 12.6° 1307

15 1 eye

22 18.2° 18.2° 23a 15.0° 14.8°

24 18.4°

Wily 14 Zi.Or 21.6 a0.Ga

21 202. 19.0

Aug. 6 21.0° 21.6°

19 25.00

20 20.6° 2As

Sept. 15 102° 18.4°

16 T7An

Oct, 2 14.6° VI Nye 13.6° 15.63

4 16,.2° 15.0°

Readings taken during 1905 and 1906 do not present any new

variations. Some of the smaller ponds not given in the table above

show temperatures as high as 25°.

For convenience these reservoirs may be divided into three types:

| (1) Those into which the mountain water is piped and which are

used for the temporary storage of drinking water; (2) those into

which the mountain water is piped, which are used for purposes of

irrigation, etc., and in which the water remains for a considerable

length of time without change; and (3) those supplied by pipes or

ditches from the streams as they leave the mountains and which also

receive a great amount of surface water during heavy rains.

In reservoirs of the first type the water is only temporarily stored,

there being a constant inflow and outflow. As a consequence, the

temperature does not rise above 15°. The water is clean and pure.

The bottom is of clean gravel or clay, the banks either of gravel or

of granite riprap. There is no plant growth at the shore or bottom.

Reservoirs belonging to this class are Colorado Springs No. 1 and

No. 2. The Colorado Springs (Mesa) reservoir will doubtless

belong to this type, but during 1905 and 1906 the new bed made the

water unfit for use and as a consequence it was allowed to remain

until it reached a high temperature. Under this condition it would

belong to the second type.

In reservoirs of the second type the water supply is the same as

84 H. L. SHANTZ

‘n that of the first. The difference is due to the greater length of

time which the water remains in these reservoirs. As far as the

writer has been able to ascertain, very little water has been taken

out for any purpose and consequently the water has risen to a tem-

perature of 21.4°. In this type there were also found differences of

shore and bottom, due to the increase in plant growth resulting

from the more favorable temperature. In the older reservoirs of

this type (Prospect) the shore is well covered with low-growing

sedges and rushes and the bottom often covered with Nitella and

Potamogeton. The younger reservoirs show very little growth

along the shore line, but the bottom is usually well covered with

Riella and Chara. This type includes Mesa Nos. I, 2 and 3, Pros-

pect, Broadmoor, and Colorado Springs (Mesa). The most favor-

able conditions for plant and animal growth are found in reservoirs

of the third type. During heavy rains great quantities of silt and

organic matter are carried into the supplying ditches and often di-

rectly into the reservoirs. The water is stored throughout the sea-

son, or very nearly, and reaches a temperature as high as 25°. The

shore line is overgrown as a rule with rushes, sedges, willows, etc.,

and the reservoir filled with growths of Potamogeton, Philotria,

Batrachium, and Chara.

The plains lakes differ from the alpine lakes in having a higher

temperature, and also in the fact that this high temperature con-

tinues for a longer period. Like the alpine lakes they are subjected

to low relative humidity of the air and consequently rapid evapora-

tion.

PLANT GROWTH

Aside from the microscopic algae, perhaps the most abundant

plant to be found in the plains lakes is Chara foetida. The flora of

the reservoirs of the third type is largely Potamogeton lucens,

together with a lesser amount of Batrachium flaccidum, and Philo-

fria angustifolia. These plants, together with Cladophora, Sptro-

gyra, and Zygnema often form a dense growth on the surface of the

pond. ‘This is especially true at Boulder and Jenks. Reservoirs of

the second type seldom show plant growth at the surface, but an

examination of the bottom usually reveals a luxuriant growth of —

Chara foetida, Potamogeton perfoliatus, Myriophyllum spicatum,

Nitella, and Riella. In reservoirs of the first type no macroscopic —

plants were found.

LAKES OF PIKE’S PEAK REGION 85

Turning now to the alpine lakes, we find in the smaller bodies, or

ponds, an abundant growth. According to Clements (1904: 353)

the most important plants in these alpine lakes are, in order of their

importance, Sparganium angustifolium and Potamogeton alpinus,

and those of secondary importance, Utricularia vulgaris, Callitriche

bifida, and Isoetes lacustris paupercula. In Isoetes Lake there is in

addition to those cited above a rather rank growth of Batrachium

flaccidum near the center of the lake, inside of a broad zone of Spar-

ganium. Michigan Lake is completely overgrown with Sparganium

angustifolium, and also shows a rather abundant growth of moss.

At Bald Mountain Lake the mosses constitute the only macroscopic

flora. The other types of alpine lakes show no flora aside from

the algae.

VERTEBRATE FAUNA

The vertebrate fauna is never abundant. Rana pipiens is present

in small numbers in the reservoirs of the third class. Amblystoma

and Bufo larvae, although present, are never abundant. Here are

also found a few fish, chiefly suckers. In reservoirs of the second

class only a few larval salamanders and suckers have been noted.

Exceptions should be made in the case of Prospect Lake, in which

tadpoles are at times very abundant and which has also been stocked

with trout. Reservoirs of the first type have no vertebrate fauna.

The most abundant vertebrate form of the alpine lakes is the

larva of Amblystoma. These were especially abundant in Mirror

and Ribbon lakes and are occasionally found in Isoetes and Michi-

gan lakes. The writer has observed a few large trout in Mirror,

Ribbon, and Isoetes lakes. In the latter they are probably not

found at the present time because of the changes in the character

of the water. These lakes are all connected with Beaver Creek,

from which mountain trout are still occasionally taken. In none

of these lakes has the writer ever seen minnows. Lake Moraine,

on the other hand, is fairly well stocked with trout and suckers, and

here are observed great numbers of minnows. Dead Lake has no

vertebrate fauna (Shantz 1905: 258).

THE PLANKTON

It is evident that in such small bodies of water no sharp line can

be drawn between the plankton, the bottom, or the littoral fauna

and flora. Even in the largest lakes a better classification would

be into plankton and littoral, since the bottom is never more than a

86 H: Li. SHANTZ

few meters from the surface, and littoral forms are always abundant

here.

Collections were taken with a Birge net, from both surface and

the bottom, and in only a few cases are the plankton forms absent

in the bottom hauls.

The alpine lakes—Mirror, Ribbon, Dead, and Moraine,—and the

lowland lakes of the first and second types——Mesa Nos. 1, 2 and 3,

Colorado Springs Nos. 1 and 2, Prospect, and to a lesser extent

Portland,—showed a plankton comparatively free from insect larvae

and other forms characteristic of bottom and shore.

There is great variation in the amount and kind of plankton

found in the different lakes. In none of the lakes studied does a

“bloom” appear. Even in the smaller ponds there is never an

abundant development of Cyanophyceae. Except for a very short

time at the end of summer the plants are never a prominent part of

the plankton.

Plankton of the Alpine Lakes

Dead Lake.—The plankton of this lake is far more abundant than

that of any other alpine lake studied. Collections taken on May 20,

1904, show a plankton almost entirely of larvae of Diaptomus

shoshone and Branchinecta coloradensis. On June 5 Daphnia was

present in small numbers and this form had increased greatly by

June 17. Diaptomus and Branchinecta were full grown on July 12.

These species are large, noticeable forms, visible to the naked eye.

The former is of a very deep red color. At this time there was also

an increase in plant growth. Cyanophyceae and diatoms were

present and Charactum was more or less abundant upon the appen-

dages of the crustacea. By July 29 the algae were more noticeable

and on August 12 they were a rather important part of the plank-

ton. By this time Branchinecta had entirely disappeared and Diap-

tomus and Daphnia were present in about equal numbers. On

August 13, 1903, and August 15, 1905, the plankton consisted

almost entirely of Diaptomus. The next collections were made on

August 26, but this time there were about three Diaptomus to one

Daphnia. A few specimens of Chydorus were also taken. On

September 17 Spirogyra and Oedogonium were present in large

numbers and these, together with other algae, constituted the greater

part of the plankton. Diaptomus still continued to be more numer-

ous than the Cladocera. The latest collections were made on Octo-

LAKES OF PIKE’S PEAK REGION 87

ber 4. This collection consisted largely of the algae, Spirogyra,

Oedogonium, and diatoms. Diaptomus still remained the most im-

portant element of the fauna, Daphnia had decreased, while Chydorus

was found in considerable numbers.

Throughout the summer months Diaptomus shoshone is abundant

at all times in the plankton of this lake. In all probability it con-

tinued for a considerable period after the last collection. Material

collected at this lake on September 17, 1904, was taken in a small

vial to Lincoln, Nebraska, placed in an icebox with a temperature

ranging from I1° to 13°, and kept in a living condition for some

time. In this vial there were two specimens of Diaptomus shoshone,

which remained alive and active until April 5, 1905, having lived

for six and one-half months in a three-drachm vial. About two

weeks later two larvae with the characteristic red color were noticed,

but these soon died. No winter collections were taken from this

lake, but it seems likely from the above experiment that this form

would be taken during the winter months.

In general, we may say that Diaptomus and Daphnia continue

throughout the season, that Branchinecta is present during the spring

_ and early summer, and that the algae become the most important

- constituent during the autumn.

Mirror and Ribbon Lakes.—A very meager plankton was found

in these lakes. Conditions for life were almost identical in the

two. Three good hauls with the net seldom gave more than

enough to cover the bottom of the vial. They were separated by

a narrow bar, about 50 m. in breadth, through which they were con-

nected by a stream from Mirror to Ribbon. |

In amount the plankton does not vary to any very great extent

from month to month. Ribbon Lake showed on May 20, 1904, a

few larval copepods, a few Daphnia, and also a few Anuraea coch-

Jearis and diatoms. On June 5 practically the whole collection was

of Diaptomus, with a very few Anuraea cochlearis and Anuraea sp.

On June 17 the only change noted was the appearance of a few

Cyclops and diatoms. Anuraea cochlearis was most important in

the plankton on July 12. Diaptomus was also present, as well as

Anuraea sp., but on July 29 Diaptomus was again more abundant

and on August I2 practically the whole collection was of Diaptomus..

An increase in the number of Diaptomus was noted August 26.

At this date there were also a small number of Daphmia and Cyclops.

88 H. L. SHANTZ

On September 17 the forms found were, in order of their importance,

Diaptomus, Daphnia, Anuraea sp., diatoms, and Zygnema. Anu-

vaca sp. was the most important form on October 4, and A. coch-

learis and Diaptomus were present in small numbers. In Mirror

Lake no Daphma were taken. This agrees with the report by

Birge (1904: 149) on Ward’s collections. In these lakes the chief

element of the plankton is Diaptomus. During June, July, and

August Anuraea was found, and in the latter part of the summer

algae,—Spirogyra, Zygnema, and diatoms,—had increased and con-

stituted a considerable part of the plankton.

Lake Moraime—tThis lake shows much less plankton than any

of the others. The vials showed no visible forms and almost no

organic fragments. A meager plankton was taken on May 20,

1904, consisting entirely of Rotifera, Asplanchna and Anuraea

being the most abundant. By June 5 Anuraea cochlearis consti-

tuted practically the whole plankton, with the exception of an occa-

sional diatom, Asplanchna, and one or two other rotifers. An in-

crease in the number of Asplanchna was noted on June 17, while

on July 12 there was a marked increase in the amount of plankton,

the dominant species being Anuraea cochlearis tecta. The plank-

ton on August 26 was by no means as abundant, there being only

an occasional Anuraea cochlearis, Pediasirum, or diatom. The last

collections made showed very little life of any kind, only an occa-

sional diatom or rotifer.

The most noticeable feature of the plankton of this lake is the

entire absence of crustacea. Very few Cladocera were taken in the

bottom hauls, but at no time were they present in the plankton. A

possible explanation of this may be found in the presence in this

lake of great numbers of minnows. This lake is also different from

the others under consideration in that the plankton is almost exclu-

sively of Rotifera and that even in autumn there is no appreciable

amount of algae developed.

The plankton of the alpine lakes is never abundant. For the

greater part of the year animals are the chief constituents. Only

late in the fall do the algae become dominant. Dead Lake shows

the richest plankton, and Lake Moraine the least amount, while

Mirror and Ribbon were intermediate as to the amount of plankton

produced. In Mirror and Ribbon the plankton was largely Diap-

tomus, in Dead Branchinecta and Daphnia also become important

'

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i

‘

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LAKES OF PIKE'S PEAK REGION 89

parts, while in Moraine the plankton is almost entirely of Rotifera.

The maximum amount of plant growth is not at the time of highest

temperature, but occurs much later.

Plankton. of the Plains Lakes

Portland.—This is the only reservoir of the third type which can

be said to have a plankton distinct, to some degree at least, from the

plant and animal life of bottom or shore. The collections on July

14 showed great numbers of Daphnia. There were also present

Anuraea cochlearis, Anuraea sp., Cyclops, and a few threads of

Spirogyra. The plankton at this time was more abundant than in

any other collection taken from this lake. By the nineteenth of

August Diaptomus was the principal form, together with Daphma,

Anuraea, Difflugia, Pediastrum, Dictyosphaerium, and Volvox.

The plankton of this collection was about one-fourth the amount of

that taken July 14, while on October 2 not more than one-eighth the

original amount was present, made up largely of Daphnia, Diap-

tomus, and diatoms. (The collection on September 2, 1903, showed

an exceedingly rich plankton, in which the following elements were

of about equal importance: Daphnia, Diaptomus, and Volvox, with

a lesser amount of Pedtastrum.) %

Mesa Reservoir No. 3—Although very young this reservoir

shows a comparatively abundant plankton. Collections made May

19, 1904, were largely of Daphnia, with a very few Cyclops. Nine-

tenths of the plankton on June 8 was Daphnia, but Diaptomus,

diatoms, and desmids also occurred in considerable numbers. The

most abundant plankton was collected June 22, when there were

present, in addition to those already cited, many larval copepods,

and also Chydorus and Cosmarium. The next collection was only

about one-third as large and was taken on July 21. Daphnia had

almost entirely disappeared; Diaptomus characterized the plankton,

but Volvox and Conochilus were also numerous, while diatoms and

Cosmarium were present in considerable numbers. The plankton

on August 20 was made up almost entirely of Diaptomus and Cono-

chilus, Volvox having disappeared. The next collection, on Sep-

tember 15, showed a decrease in the animal and an increase in the

plant constituents of the plankton. The forms found were, in the

order of their importance, diatoms, Oedogonium, Diaptomus, Chy-

dorus, another cladoceran, Cosmarium, and Difflugia. The latest

/

go H. L. SHANTZ

collections were on October 4, and showed Diaptomus, Conochilus,

Daphnia, diatoms, and filamentous algae.

It will be seen from the above that Daphnia dominates in the

spring plankton, Diaptomus in the summer, and Conochilus and

algae late in the summer and autumn. At the very end of the season

there was again an increase in crustacea.

Mesa No. 1.—The earliest collections from this reservoir were on

May 19, 1904. At this time there was almost as much plankton as

at any period during the year. Chydorus, Daphnia, Diaptomus, and

copepod larvae were present in about equal amounts. There was a

considerable amount of Mesocarpus. Daphnia and Diaptomus were

about equal on June 22, at which time Chydorus and several ostra-

cods were taken. The plankton was more abundant on July 21

than at any other time, and was made up of the same constituents

as noted above. On August 20 Cladocera had entirely disappeared,

the bulk of the plankton being of Diaptomus and of Conochilus. A

marked decrease in the amount was noted on September 15. Diap-

fomus, diatoms, and two Daphnia were found, while on October 4

the plankton was made up of a very small number of diatoms.

Collections made during 1903 and 1905 do not show any marked

variation from those of 1904. Mesa No. 2, in a general way, shows

very little variation from that of No. 1.

Prospect Lake.—This lake has never yielded as much plankton as

Mesa Nos. 1, 2 or 3, or Portland. On May 27, 1904, Cyclops, dia-

toms, rotifers, and Chydorus constituted the very meager plankton.

Bosmina was by far the most important element of the plankton taken

June 24 and July 14. Cyclops, diatoms, Cyanophyceae, and Anu-

raea cochlearis, and several rotifers were present. The most abun-

dant plankton ever taken from this lake was collected August 6,

1904. It was almost entirely of Bosmina with a few Cyclops,

Anuraea, Rattulus, and Peridinium. The character of the plankton

had greatly changed on October 2. The forms, in order of their

importance, were Peridinium, Bosmina, copepod larvae, diatoms,

Pediastrum, Merismopedia, Staurastrum, Cosmarium, Oscillatoria,

and Cyclops. Although the last collection was made up of a number

of elements the total amount was very small.

The plankton of this lake contrasts sharply with that of the

other lakes in the fact that here for the first time Bosmina is found.

All of the lakes considered up to this point, with the single excep-

LAKES OF PIKE’S PEAK REGION QI

tion of Lake Moraine, show a plankton dominated by Diaptomus.

In this lake, however, Diaptomus is almost entirely lacking. Cyclops

is present but, with the exception of the last collection and that taken

May 27, Bosmina was at all times the most important element. The

presence of Peridinium and its dominance in the October collection

is also a notable feature.

Colorado Springs Reservoir No. 2.—Before considering the

plankton of this reservoir it will be well to mention one or two facts

with respect to the water supply. The water which supplies this

reservoir passes first through the pipes of the Colorado Springs

water system. These go direct to the consumer, and it is only the

excess of water which passes into this reservoir or reservoir No. I.

While these reservoirs are continually being supplied with water

from the pipes, they are also being drawn upon by the consumer.

In consequence, the water may remain in the reservoir for only a

few hours. This fact alone is probably sufficient to explain the very

meager plankton found here. Many collections show absolutely no

living forms, but occasionally an organism will be taken. About the

only plankton taken from reservoir No. 2 was on June 8, when a

very few rotifers were present, as well as one larval copepod. On

June 22 no organism was found, but on October 2 a few specimens

of Peridinium were noted in the collection.

Reservoir No. 1.—The collection on June 8, 1904, in this reservoir,

as well as that on June 22, showed no organisms. The most abun-

dant plankton was found on October 2. Here the collection showed

an occasional Peridinium, Merismopedia, or rotifer. The bottom

collection on this date showed a rather large number of Merismo-

pedia, with a few diatoms, rotifers, and insect larvae.

It will be remembered that the water supplying these two reser-

voirs comes originally from the intake in Bear Creek and Ruxton.

The water supply at Bear Creek is from a mountain stream, in

which plankton forms would not be found. That supplied from the

intake on Ruxton has come from Lake Moraine, a distance of about

three miles in an open stream. It will also be remembered that

Lake Moraine showed almost no plankton aside from the Rotifera.

Under these conditions one would not expect plankton to develop

in reservoirs Nos. 1 and 2.

92 H. L. SHANTZ

LITTORAL FLORA AND FAUNA

Here it seems best to include the animal and plant forms which

occur in such numbers in the mountain ponds and the plains reser-

voirs of the third type.

Bald Mountain Lake.—Of the mountain ponds perhaps the most

interesting, because of its great altitude, is that found on Bald Moun-

tain. Collections from this pond made June 17, 1904, at which time

ice still covered the pond, were largely of Cyclops, Chydorus, and

nematodes. There were also many diatoms and insect larvae as well

as occasional rotifers. Dinobryon and Macrobiotus macronyx were

also noted. July 12 showed Dinobryon, diatoms, Zygnema, Meso-

carpus, as well as copepod larvae. There were also noted Macro-

thrix montana and Chydorus. On August 12 Mesocarpus and

Zygnema, together with a number of the Cyanophyceae, constituted

the principal part of the flora, while the fauna was almost entirely

of Chydorus, Cyclops, and Rotifera. Life was not as abundant on

August 26, when larvae, diatoms, Gomphiosphaerium, and Cyclops

were the principal forms. The September 17 collection showed an

abundance of algae, chiefly Mesocarpus and Zygnema, filamentous

and other diatoms, and Gompluosphaerium. Isopods, rotifers and

copepod larvae were also taken. Collections on August 15, 1905,

showed a number of Ostracoda in addition to those already men-

tioned.

Michigan Lake.—Of the alpine lakes Michigan shows the greatest

abundance of living forms. ‘The earliest collections, made on May

20, show chiefly rotifers. Dinobryon was the most important form

on June 17, while on July 12 a very few Cyclops and Diaptomus

were taken. On this date Dimobryon had disappeared. On July

29 Daphnia was the most abundant form, Anuraea cochlearis was

next in importance, and Cyclops was also found. A very few speci-

mens of Bosmina were taken in this lake. The collection on August

12 was much richer than any of those previously taken. Diaptomus,

Daphma, and Anuraea cochlearis were the most abundant. About

the same forms were found on August 26, but there was consid-

erable increase in the amount of copepod life. Algae were dominant

on September 17, chiefly Spirogyra, and a lesser amount of Zygnema

and diatoms. Of animal forms, Daphnia and Diaptomus were of

the greatest importance. The latest collections were made on Octo-

ber 4. These were almost entirely of Spirogyra, with a consider-

able number of Daphnia and Diaptomus. Insect larvae were abun-

dant in all of the collections.

LAKES OF PIKE’S PEAK REGION 93

Tsoetes Lake.—In this lake the collections on May 20, June 5 and

17, July 12 and 29, showed only an occasional rotifer, Diaptomus,

Dinobryon, diatom, or insect larva. The collections were largely of

silt and organic remains. On August 12, however, Chydorus was

rather abundant, as were also Rotifera, Dinobryon, diatoms, and

insect larvae. The collection on August 26 was largely Diaptomus,

Chydorus, Cyclops, and Anuraea. On September 17, and again on

October 4, a small amount of Spirogyra and Zygnema made up the

ereater part of the haul.

Although the conditions in these alpine ponds seem especially

favorable for growth, it will be seen from the above that in them

life is not abundant during the early part of the summer period.

During the latter part of the summer and continuing until the

approach of winter the algae are developed in considerable num-

bers, in some cases filling up the pond almost completely.

Boulder Lake.-—Of the lowland lakes of the third type, Boulder

is perhaps the best example. A collection made here on May 27,

1904, showed only a few larval copepods. On June 23, however,

Cladocera were abundant, chiefly Bosmina. Cyclops also was rather

abundant at this time. Collections on June 14 were especially rich

in Diaptomus, Daphnia, other Cladocera, Cyclops, Ostracoda, Spiro-

gyra, Oedogonium, and insect larvae. The same might be said to

be true of the collection on August 6, except that the insect larvae,

Diaptomus, and Ostracoda were more abundant. The richest col-

lections were taken on September 16, when the surface showed

chiefly Daphnia, Diaptomus, Cyclops, Anuraea, and many algae.

The collections on October 2 did not differ markedly from those of

September 15. This pond also showed Volvox, C oleochaete, Bulbo-

chaete, and Cladophora.

In reservoirs of the second type the abundant growth of Chara

and other macroscopic plants insures an abundance of insect larvae,

Cladocera, diatoms, and desmids. Without exception all of the

plankton forms were also taken from the bottom. Colorado Springs

reservoirs Nos. 1 and 2 showed a very little growth of any kind

from bottom hauls. Merismopedia, an occasional diatom, rotifer,

or insect larva were the elements of the bottom hauls.

Of the alpine lakes, exclusive of the ponds, Dead Lake showed

the most abundant life at the bottom. In addition to the plankton

forms already mentioned insect larvae and Turbellaria were most

94 H. L, SHANTZ

abundant. From this bottom one small bivalve mollusk was taken,

this being the only bivalve mollusk found by the writer in any of

the lakes under consideration.

Bottom hauls from Mirror and Ribbon lakes did not differ essen-

tially from the plankton hauls except in the greater abundance of

diatoms.

RESERVOIR No. 5

This reservoir covers the old sites of Mirror, Ribbon and Marsh

lakes, as well as a considerable atnount of mountain meadow and

thicket region lying near them. It was completed in 1904, but it

was not until the middle of the summer of 1905 that this reservoir

was filled. All shrubs lying within the basin were cut down and

the roots and plant remains removed in so far as possible. In many

places all surface vegetation was removed, leaving a clean gravel

bottom. Near the upper end of the reservoir and also along many

of the sides, the water has spread out over the thick mat of mountain

meadow. ‘This meadow is a dense growth of grasses and herbs,

which never rise more than a few centimeters above the surface of

the soil and form a thick mat-like layer. The old bottoms at

Mirror and Ribbon lakes have been left undisturbed. Marsh Lake,

on the other hand, was completely drained and the bottom scraped

out and lowered several meters. |

We may distinguish three types of bottom in this reservoir: First,

the original clay bottoms of Mirror and Ribbon lakes: second, the

clean soil or gravel bottom; and, third, the submerged mountain

meadow.

On July 14, 1906, water was turned into this large reservoir and

by August 21 the lakes had been completely covered. On this date

the water level over the former Mirror Lake was about 5 m. above

that of the old lake surface. According to the report of the man in

charge of this reservoir the water was rising at this time at the rate —

of about 3.8 cm. each day. At the edge of the reservoir and extend-

ing in to the depth of a meter or more could be seen many flowering

plants and fungi, which had not yet decayed as the result of having

been covered with water. |

The water supply of this reservoir comes from the stream leading

from Windy Point on Pike’s Peak and is consequently free from

plankton forms. In the old basins of Mirror and Ribbon lakes there

still remained a large amount of water, but this, by the addition of

LAKES OF PIKE'S PEAK REGION 95

water from the mountain streams, was diluted many times. This

would naturally result in a great decrease in the amount of plank-

ton in proportion to the amount of surface.

Collections were made in this reservoir on August 21 and Sep-

tember 24, at which time the temperature was 15°C. No very

noticeable difference was found in the plankton of these two dates,

with the exception that the algae were somewhat more abundant on

the latter date. At this time plankton taken over the old bed of

Ribbon Lake showed Daphnia, Diaptomus, Anuraea cochlearis, and

several other rotifers. Unicellular algae were also rather abundant

in this collection. At the same time a collection over the Mirror

Lake basin showed Daphnia, Diaptomus, Chydorus, Mesocarpus,-

Zygnema, Anuraea cochlearis, and Dinocharts. This plankton was

much more abundant than that taken over the former Ribbon Lake.

Several reasons may be given for the abundance of the plankton

over the old Mirror Lake basin. The wind usually blows from the

former to the latter and this would carry with it the greater part

of the plankton. Around the old bed of Mirror Lake there is sub-

merged a large area of mountain meadow; this would mean a rich

supply of food for both plant and animal life in the region adjacent

to it. .

At the lower end of the reservoir, at the place where. Marsh Lake

was formerly located, Anuraea cochlearis and an occasional Diap-

tomus or Cyclops constituted the entire plankton. Plankton taken

on the new gravel bottom was even less abundant than that taken

from over the former Marsh Lake, although made up of the same

elements. The conditions over the latter area differ very little from

those over the new bottom, because of the fact that the old bed of

this lake was entirely removed.

The most abundant plankton was taken over submerged mountain

meadow. Here there was an abundance of Mesocarpus and Zyg-

nema, Diaptomus, desmids, and rotifers. In addition to these forms

there was also present a great amount of plant fragments, chiefly

anthers, pollen, etc.

The life at the bottom of the old lake beds had changed very

little. The two principal changes in habitat, increase in food and

increase in depth, had operated for only a short time and the changes

were only a very slight increase in the amount of life, chiefly algae.

On the gravel bottoms living forms were hardly to be found, but

96 H. L. SHANTZ

on the old submerged meadow life was very abundant. The chief

growth was of the algae, Zygnema and Mesocarpus. The animal life

was almost entirely Diaptomus, Daphnia, Chydorus, Anuraea, and

other rotifers.

By comparing the collections taken from these lakes in 1904 and

1905 with those taken from the reservoir in 1906 a great increase

is noticed in the amount of plankton. There has also been an in-

crease in the number of species and genera represented. ‘This result

would naturally be expected when we consider the conditions of

the two regions. The old lake beds of Mirror and Ribbon lakes had

particularly clean bottoms and shore lines. There was no notice-

able plant growth and the microscope revealed only a few diatoms

and occasional filaments of algae. The reservoir, on the other hand,

has a bottom largely made up of abundant plant and animal remains

and these form a most abundant food supply for both plants and

animals.

COMPARISON OF PLAINS LAKES AND ALPINE LAKES

In comparing the life of the plains lakes with that of the mountain

lakes, the most striking thing is the greater abundance of life in the

plains lakes. The plankton collections from these lakes contain

many times the amount of material shown in collections from the

alpine lakes. Exception should be made in the case of Prospect

Lake, which at no time gave more plankton than did Dead Lake.

This is also true of Colorado Springs reservoirs Nos. 1 and 2, but

this condition is explained by the peculiar conditions of water supply.

In a very general way the plankton of Lake Moraine resembles

that of the two reservoirs just mentioned. It is very meager in

each case and is largely of Rotifera. The alpine lakes, Mirror,

Ribbon, and Dead, were or are dominated by Diaptomus and Daph-

nia, and are to some extent comparable to Mesa reservoirs Nos. 1,

2 and 3, and to Portland Lake, which are dominated by species of.

the same genera. Prospect and Boulder lakes are alike in having

the plankton largely of Bosmina. Portland Lake, a very young

reservoir, will in a few years approach more closely to Boulder

Lake, and Mesa reservoirs Nos. 1, 2 and 3 will in time become like

Prospect Lake. 3

ACKNOWLEDGMENTS

The writer wishes to express his appreciation of the assistance

given by Professor Henry B. Ward. The study of these lakes was

LAKES OF PIKE'S PEAK REGION 97

suggested by him and for two years was carried on under his super-

vision. Acknowledgment should also be made of the assistance

given by Mr. H. I. Reed, civil and consulting engineer, of Colorado

Springs, Colorado, who has kindly furnished much of the data on

the age, area, and depth of the reservoirs under consideration.

PAPERS CITED

Birce, E. A.

1904. Report on the Cladocera (in Ward, 1904). Trans. Amer. Mic.

Soc., XXV: 149-151.

CLEMENTS, F. E.

1904. Formation and succession Herbaria. Univ. Neb. Studies, IV:

329-355: :

SHANTZ, H, 1:

1905. Notes on the North American species of Branchinecta and their

Habitats. (Studies from the Zoological Laboratory, The University

of Nebr., No. 62.) Biol. Bull, IX (4): 249-264, Pls. X-XII.

Warp, H. B.

1904. A Biological Reconnaissance of some Elevated Lakes of the

Sierras and the Rockies. (Studies from the Zoological Laboratory,

The University of Nebraska, No. 60.) Trans. Amer. Mic. Soc.,

XXV: 127-154, 12 pls.

ZSCHOKKE, F. i

1900. Die Tierwelt der Hochgebirgseen. Neu. Denkschr. allg. schweiz.

Ges. ges. Nat., XXXVII. 400 Pp., 8 pls., 4 maps.

98 H. L. SHANTZ

EXPLANATION OF PLATES

Plate V

A map of the Pike’s Peak Region showing the relative position of the

principal lakes and reservoirs. The lakes and reservoirs are relatively large

in comparison with the other features shown.

Plate VI

Seven Lakes as seen formerly from Bald Mountain. 1, Lake of the

Rocks; 2, Ramona Lake; 3, Michigan Lake; 4, Isoetes Lake; 5, Marsh Lake;

6, Ribbon Lake; 7, Mirror Lake. (Taken by permission from Ward, 1904,

Pl, XXVI; photographed by Professor F. E. Clements in 1899. )

Plate VII

A view of Reservoir No. 5 from Bald Mountain. The region shown is

the same as that shown in Plate VI and the members are the same as for

that plate. Marsh Lake, Ribbon Lake and Mirror Lake form one reservoir,

while a second one (Reservoir No. 4) has been formed, marked 8 in the

plate. (Photograph by Professor F. E. Clements, Aug. 28, 19006.)

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PLATE VIL

VARIATIONS IN THE VITELLARIA AND VITELLINE

DUCTS OF THREE DISTOMES OF THE GENUS

OPISTHORCHIS

By F. D. BARKER

WITH TWO PLATES

In an investigation of the variations in the various organs of

Opisthorchis pseudofelineus Ward, one is particularly impressed

with the interesting and striking variations which occur in the

vitellaria and vitelline ducts of this species. These variations are

described in the following paper and for the purpose of comparison,

the results of the work of two other investigators, Weski (1900)

and Muhling (1896), are also given. Weski and Muhling have

worked out the variations in the vitellaria and vitelline ducts of two

species of the genus Opisthorchis, closely related to O. pseudo-

felineus.

Ward (1895: 241) describes the vitellaria of Opisthorchis pseu-

dofelineus as follows: “The glands begin about half as far behind

the acetabulum as that is from the oral sucker, and extend to the

middle of the space between the two testes, or even sometimes as

far as the anterior edge of the posterior testis. One break in the

line of acini may always be recognized as most prominent; it is

located just opposite the ovary on each side, and is in length more

or less equal to the diameter of the ovary. This space divides the

vitellarium into two portions, which may be distinguished as anto-

varial and postovarial. While it is in some cases possible to dis-

tinguish in the antovarial portion groups of acini, they seem to be

usually rather indistinct, or at least very unequal in size, as if adja-

cent groups had become confluent by the growth of interlying acini.

The postovarial portion, however, is usually distinctly divided into

two or three groups of acini, though even these may be obliterated.

In two or three cases a small group of acini was found, on one

side only, in this intermediate space opposite the Ovary, and was

I0O F. D. BARKER

clearly separated from both antovarial and postovarial portions by

a small space. Corresponding to the two portions of the gland, one

Gnds on each side two ducts which, extending obliquely toward

the ovary, from a short distance before and behind it, form a ‘Y’

or ‘V’ according as they meet before or not until after reaching

the ovary. These ducts on either side of the ovary form one of the

most characteristic appearances of the specimen.” Stiles (1904:

32) gives the following description of these glands: “ Vitellaria

divided each side into an antovarial portion with about five acini

and a postovarial portion with two or three acini, each portion

provided with its own vitelloduct ; the antovarial portion extends

from the ovary cephalad to about the boundary between the anterior

and equatorial thirds of the body; the postovarial portion extends

caudad from the ovary to about the anterior plane of the posterior

testicle.”

Looss (1899: 675) considers the vitellaria the most constant spe-

cific character, not only in the genus Opisthorchis, but also in all

distomes, and holds that the form and size of the other genital

organs are of much less value as specific characters. It is of great

importance to determine in specific cases just how far this view 1s

correct.

The results of the examination of more than one hundred speci-

mens of Opisthorchis pseudofelineus show that there is considerable

variation even in the vitellaria, and make a few slight changes in

the descriptions given by Ward and Stiles seem advisable.

The extent of the vitellaria, caudad and cephalad, is one of the

most important characters of the vitellaria, if not the most impor-

tant, and is of special significance as a specific character. Because

of the specific value attributed to the extent of the vitellaria, we

should expect to find their limits rather definite and constant; on

the contrary, we find that they vary considerably. The vitellaria.

never extend to the acetabulum, but their position posterior to the

posterior margin of the acetabulum on the right side varies from

0.3 mm. to 4.5 mm., the mode being 1.10 mm., with a frequency

of 23 or 23 per cent. On the left side the position varies from 0.3

mm. to 3.7 mm., the mode being 0.7 mm., with a frequency of 18

or 18 per cent. The class 1.1 mm. had a frequency of 12, the class

1.3 mm. of 14, and the class 1.5 mm. of 15. The modes for the

right and left sides show that the vitellaria extend farther cephalad

VARIATIONS IN VITELLARIA AND VITELLINE DUCTS 1OL

on the left than on the right side. In individual specimens the

extent of the vitellaria cephalad may be equal on the right and left

sides or greater on the right than on the left side.

A comparison of the length of the anterior portion of the body

(from the anterior end to the ovary) with the position of the vitel-

laria, posterior to the ventral sucker, shows that the extent of the

vitellaria cephalad varies independently and is not affected by the

degree of contraction of the anterior portion of the body.

In three living specimens which were observed, those regions of

‘the worm anterior to the acetabulum and posterior to the ovary

were found to be most distensible and consequently to vary most in

length with the contraction and expansion of the worm. In view

of these facts Ward’s (1895) description of the anterior limits of

the vitellaria, “the glands begin about half as far behind the ace-

tabulum as that is from the oral sucker,” and Stiles’ (1904) de-

scription, “the antovarial portion extends from the ovary cephalad

to about the boundary between the anterior and equatorial thirds of

the body,” are not sufficiently definite, since the relative position of

the acetabulum and the oral sucker and the boundary between the

anterior and equatorial thirds of the body will vary with the con-

traction and expansion of the anterior region of the body. The

limits of the vitellaria cephalad, though they vary with reference

to the posterior margin of the acetabulum, are more definite if

given in millimeters, inasmuch as that portion of the body posterior to

the acetabulum is not so much affected: by the state of contraction

or expansion of the anterior region as that portion of the body

anterior to the acetabulum.

It is convenient to state the extent of the postovarial portion of

the vitellaria with reference to the testes and inasmuch as the rela-

ive position of these organs is fairly constant, such a designation is

sufficiently definite.

The extent of the vitellaria caudad varies from the anterior

margin of the anterior testis to a plane 0.07 mm. to 0.30 mm. pos-

terior to the posterior testis, the mode being the anterior margin of

the posterior testis, with a frequency of 28 or 28 per cent. for the

right side.

On the left side the extremes were found to be a plane 0.16 mm.

anterior to the anterior testis and a plane 0.07 mm. to 0.25 mm.

posterior to the posterior testis, the mode being the anterior margin

102 F. D. BARKER

of the posterior testis, with a frequency of 22 or 22 per cent. The

planes and margins used in designating the limits of the vitellaria

are the transverse planes of the body of the worm. |

The extent of the vitellaria caudad is, in the aggregate, approxi-

mately the same for the right and left sides. The vitellaria extended

to or beyond the anterior margin of the posterior testis in seventy-

one specimens on the left side and in seventy specimens on the right

side. In 12 per cent. of the specimens the vitellaria extended to the

posterior margin of the posterior testis on the left side and in 9 per

cent. on the right side. In 8 per cent. the vitellaria extended from

0.07 mm. to 0.25 mm. beyond the posterior testis on the left side

and in 7 per cent. from 0.07 mm. to 0.30 mm. beyond it on the

right side. The extent of the vitellaria may be equal on both sides,

as was the case in 27 per cent. of the specimens, or the extent may

be greater on the left side than on the right side, as found in 38 per

cent., or the extent may be greater on the right than on the left

side, as found in 38 per cent.

The posterior third of the body was found to be the most dis-

tensible in living specimens and consequently the position of the

posterior testis and the posterior limits of the vitellaria vary greatly

with reference to the posterior margin of the worm, but the rela-

tive position of the testes and the limits of the vitellaria remain very

constant.

The occurrence, extent, and position of the break between the

antovarial and postovarial portions of the vitellaria were found to

vary considerably. On the right side, in two specimens, no break

occurred, the last group of the antovarial portion and the first group

of the postovarial portion being in contact, but not connected by a

duct. This complete obliteration of the break is evidently not due

to the contraction of the worm, for in both of these specimens there

was a distinct break on the opposite side (pl. vim, fig. 11). In .

ten specimens a distinct break occurred, but the two regions were

connected by a vitelline duct similar to the ducts which connect the

groups of either region (pl. vii, figs. 4, 7, 9).

On the left side the antovarial and postovarial regions were in

contact in one specimen and connected by a vitelline duct in four

specimens. The extent of the break between the postovarial and

the antovarial portions was found to vary from 0.08 mm. to 1.3 mm.

on the right side, 0.2 mm. being the mode, with a frequency of 33

VARIATIONS IN VITELLARIA AND VITELLINE DUCTS 103

or 33 per cent. On the left side the extremes were 0.06 mm. and

1.9 mm., the mode being 0.2 mm., with a frequency of 26 or 26

per cent. In nine specimens the break was less than 0.1 mm., this

being less than the average break between the groups of either

the antovarial or the postovarial region. The middle third of the

body is least affected by the contraction and distension of the worm.

A comparison of the extent of this break with the length of the

worm shows that the extent of the break is not affected by the

degree of contraction of the worm, but that it varies independently.

The position of this break in relation to the ovary varies some-

what, but is more constant than the characters already mentioned.

The extremes for the position of the break were found to be a

position more or less anterior to the anterior margin of the ovary,—

in one specimen (pl. vil, fig. 3) far anterior,—and a position more

or less posterior to the posterior margin of the ovary, with all grada-

tions between these extremes. The mode was a position opposite

the middle of the ovary, with a frequency of 49 per cent. on the

right side and 55 per cent. on the left side.

The distinctness and consequently the number and size of groups

in the antovarial and postovarial region varies considerably. In

some specimens the groups appear to be fused and form one or more

continuous masses. As Ward (1894) suggests, this is probably due

to a confluence of close-lying groups or the growth of intermediate

groups. The groups are more distinct and the number more definite .

in the postovarial regions than in the antovarial regions.

In the specimens in which the groups were definite the number

varied from one to six groups in the right antovarial region, five

groups being the mode, with a frequency of 61 per cent., and either

two or three groups in the right postovarial region, there being an

equal number having the two groups and the three groups. The

combination of five groups in the antovarial and three groups in

the postovarial region was found to be the most common, 30 per

cent. having this combination.

On the left side the number of groups varied from one to six

in the antovarial region, 67 per cent. having five groups. In the

postovarial region the number of groups was either two or three,

58 per cent. having three groups. For the left side the most fre-

quent combination was found to be five groups in the antovarial

and three groups in the postovarial region, 43 per cent. having this

104 F. D. BARKER

combination. The arrangement of groups is seen to be more con-

stant on the left than on the right side.

In eight specimens a single group, which Ward (1894) mentions,

was found in the space between the antovarial and postovarial por-

tions of the vitellaria and distinctly separated from both portions

by a small space. This intermediate group occurred in two speci-

mens on the right side and in six specimens on the left, never occur-

ring on both right and left sides in the same specimen and was in

every case the sixth group counting from the anterior (pl. vit,

figs. 5, 8).

In one specimen there were eight distinct and normal groups,

three groups in the postovarial and five groups in the antovarial

region, on the left side, and only five distinct and normal groups,

two groups in the postovarial and three groups in the antovarial

region, on the right side. The two anterior groups of the antovarial

region on the right side were vestigial; the first group was simply

a mass of yolk cells, with no distinct acini; the second group con-

sisted of a few acini arranged along the longitudinal duct. The

longitudinal duct was distinct and normal (pl. rx, fig. 4).

In another specimen there were on the right side four or six

groups, two or three groups (the last group probably being two

groups lying very close together) in the postovarial region and

two or three groups in the antovarial region, and only four groups

on the left, three groups in the postovarial region and one group in

the antovarial. The longitudinal duct, filled at intervals along its

course with yolk cells, extended as far cephalad as the gland on the

right side, but the groups of acini were entirely lacking (pl. Ix,

fig. 2).

In one specimen the third group of the postovarial region on the

left side was vestigial, the longitudinal duct alone remaining.

One of the most characteristic features of this species is found

in the vitelline ducts, which extend obliquely toward the ovary

from each portion of the vitellaria. These ducts form a “V” or

a “Y” as they meet after or before reaching the ovary. These

types combine in several ways to form four distinct classes. First,

a “ Y” type of duct on both the right and left sides (pl. vit, fig.

I); second, a “V” type of duct on both sides (pl. vitt, fig. 2);

third, a “ V” type of duct on the left side and a “ Y” type on the

right side (pl. vitz, fig. 4); fourth, a “V” type on the right and

VARIATIONS IN VITELLARIA AND VITELLINE DUCTS 105

a “ Y” type of duct on the left side (pl. vit, fig. 6). The follow-

ing table shows the occurrence of these various classes in one hun-

dred specimens:

Class Number of Specimens

Visi type | tight’ andi trtypededty oa poe ae ey Co es

Wxgetepe wright: ands Visite wlettsiicias ek Wie de! (lah een

PiVistypen. tight. ancivae Vaumauine verti li orice al i ukaal na eR nS

Peay pe TATE! ATICL Sy MAM EV DEELEY hones Oe dale aie ae ean 18

The study of these ducts revealed some striking variations. In

those specimens in which there was an intermediate group between

the antovarial and. postovarial regions of the vitellaria this group

receives a branch vitelline duct from the principal antovarial duct

in four specimens (pl. vii, figs. 2, 5), and from the postovarial

duct in two specimens. In one specimen the vitelline ducts were

three in number, of which the median duct passed to this interme-

diate group (pl. vit, fig. 8). |

In three specimens a short branch duct passed caudad from the

right antovarial vitelline duct, immediately after it left the group,

to the first group of the postovarial region (pl. vit, fig. 6). A

slight shifting in the position of the origin of this branch duct

would bring about the condition, which has already been mentioned,

viz., the connection by a duct of the antovarial and postovarial

regions of the vitellaria (pl. vit, fig. 9).

In one specimen the postovarial duct connected with this inter-

mediate duct instead of passing backward to the first postovarial

group (pl. vim, fig. 7). In one specimen an accessory duct was

found which connected the antovarial and postovarial ducts (pl.

vill, fig. 10). In four specimens, only one duct was found on

one side, in two cases on the right side and in two cases on the left

side. This condition is possibly due to an extreme variation of the

“Y” type of duct, in which the arms of the “ Y” form right angles,

instead of acute angles, with the stem of the “Y,” in this way

producing a “ T” type of duct (pl. vit, figs. 3, 11, 12). One finds

all possible degrees of variation from one extreme, the “ T” type

of duct passing through the “ Y ” type, to the “ V” type, the other

extreme (pl. vill, figs. 12, 13, 14, I5).

In summarizing the above discussion we find, for O. pseudo-

felineus, that the position of the break between the antovarial and

the postovarial regions of the vitellaria is one of the most constant

106 F. D. BARKER

characters; that the extent of the vitellaria caudad is extremely

variable, and therefore a characteristic which in itself is of very

little specific value; that the number of groups in each region of

the vitellaria is fairly constant, five groups in the antovarial and

three groups in the postovarial region, though variations may occur

through the fusion of groups and the appearance of accessory

groups; that the “V” type of vitelline duct predominates.

It is of value to compare with the variations noted the variations

‘n the vitellaria of two other species of the genus Opisthorchis,

which are closely related to Opisthorchis pseudofelineus and closely

resemble it in appearance and in the arrangement of their various

organs. Miuhling (1896: 260) describes the vitellaria and ducts of

Opisthorchis felineus Riv. as follows: “The vitellaria lie in the

middle third of the body, lateral to the intestinal crura and consist

generally of eight groups of small transversely arranged acini. One

rarely finds nine or seven such groups. The acini (groups) are

connected by a longitudinal duct. The paired vitelline ducts gen-

erally arise from the longitudinal duct, which connects the last two

eroups of the vitellaria, and pass obliquely backward to the ovary,

uniting in the median line to form a single short duct” (pl. 1x,

fig. 3).

Mithling (1896: 261) examined sixty specimens of O pisthorchis

felineus and found the following variations in the vitellaria and

ducts. In fifty-four cases, or 90 per cent. of the number examined,

the vitellaria were composed of eight acini (groups) ; in one speci-

men there were nine acini (groups) on the right side and in another

specimen there were six acini (groups) on each side; in five speci-

mens there were six acini on each side, but in one of these specimens

the first acinus was lacking on one side, but the longitudinal duct,

filled with yolk cells, was still present.

In 88 per cent. of the specimens the vitelline duct arose from the

longitudinal duct connecting the last two groups; in nine cases the

vitelline duct arose from the longitudinal duct which connects the

next to the last groups, namely, the sixth and seventh; in four cases

the vitelline duct arose from the sixth group; and in one specimen

the two vitelline ducts, one of which arose from the connecting duct

between the last two groups, the other duct arising. from the last

group, coalesced before passing to the ovary.

In comparing the vitellaria of Opisthorchis pseudofelineus and

VARIATIONS IN VITELLARIA AND VITELLINE DUCTS 107

Opisthorchis felineus, one notes that there is considerable difference

in the general appearance of the glands. These differences would

eliminate certain variations in the one that are found in the other.

On the whole, there seems to be less variation in the glands and

ducts of Opisthorchis felineus than in Opisthorchis pseudofelineus.

In certain features the vitellaria of Opisthorchis lancea Dies. are

very similar to the glands of Opisthorchis pseudofelineus. Weski

(1900: 580) describes the glands of Opisthorchis lancea as follows:

“The paired vitellaria lie on either side, lateral to the intestinal

crura, and extend from a plane back of the acetabulum, equal to

the diameter of the acetabulum, to the end of the intestinal crura.

They are composed of small, transversely placed acini, which appear

in more or less distinct groups. On each side there are eight

groups, which are divided into two distinct regions, an anterior and

a posterior region, by a large break between the fourth and fifth

groups. The groups of each half are connected by a median canal,

the union of which forms the transverse vitelline duct” (pl. 1x,

fig. 1).

Weski (1900) examined a large number of specimens (400) of

Opisthorchis lancea Dies. and reports the following variations in the

vitellaria and ducts.

In 158 cases there were two regions, each region having four

groups, or eight groups in all, on the right side. In 116 cases

these same conditions were found on the left side. In 22 cases

the groups were distinct, but there was no break between the ante-

rior and posterior regions on the right side, and this same condition

was found in four cases on the left side. In 119 cases the groups

coalesced on the right side and in 35 cases on the left side and no

break occurred between regions on either side. In 71 cases the

groups coalesced on the right side but the break between regions

was distinct ; this condition occurred on the left side in 245 cases.

The break occurred between the fifth and sixth groups instead of

between the fourth and fifth in 21 cases. In nine cases there were

only seven groups on the right side, the break occurring between

the fourth and fifth groups. Weski also states that there is gen-

erally a difference in the extent of the caudad half of the glands on

the right and left sides, the caudal half of the right gland gener-

ally being the shorter.

The extent and similarity in the variations of the vitellaria and

108 F. D. BARKER

ducts of Opisthorchis pseudofelineus and Optsthorchis lancea are

very striking, this condition probably being due to the great simi-

larity in the general appearance and normal condition of these

organs in the two species. One does not find such similarity in the

variations of the position of the other genital organs of these two —

species; this will be fully discussed in a paper, nearly ready for pub-

lication, on “ Variations in the position of the genital organs of

trematodes. |

I desire here to acknowledge my indebtedness and appreciation to

Professor Henry B. Ward, the Director of the Zoological Labora- :

tory, the University of Nebraska, at whose suggestion I began this |

investigation and with whose helpful co-operation I have been able :

|

.

to complete it.

PAPERS CITED

Looss, A.

1899. Weitere Beitrage zur Kenntniss der Trematoden-Fauna Aegyptens.

Zool. Jahr., Syst., XII: 521-784, Pl. 24-32.

MUBLING, P. )

1896. Beitrage zur Kenntniss der Trematoden. Arch, fiir Naturgesch., —

LXII (1): 243-276, Taf. XVI-XIX. J

STILES, CHAS. W.

1904. Illustrated Key to the Trematode Parasites of Man. Bull. No. 17,

Hyg. Lab., U. S. Pub. Health and Mar.-Hosp. Serv.

Warp, Henry B.

1895. The Parasitic Worms of Man and the Domestic Animals. Ann.

Rep. Nebr. State Bd. Agric., 1894: 225-348, text-figs. 1-81, 2 Pls.

WESKI, O. .

1900. Mitteilungen iber Distomum lancea Dies. Centralbl. f. Bakt., etc.,

XXVII (Abt. 1): 579-583.

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PLATE VIII

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VARIATIONS IN VITELLARIA AND VITELLINE DUCTS 109

EXPLANATION OF PLATES

ABBREVIATIONS

ac. Acetabulum. p. b. Pharynx.

es. Esophagus. r. s. Receptaculum seminis.

ex. c. Excretory canal. Ss. g.., Shell gland.

ex. p. Excretory pore. t. Testis.

g. p. Genital pore. ut. Uterus.

4. Intestine. vd. Vagina.

LT. ¢c., Laurer’s canal. v. d. Vas deferens.

mM. Mouth. v. dt. Vitelline duct.

o. s. Oral sucker. v. e. Vas efferens.

ov. Ovary. v. g. Vitelline gland.

Plate VIII

Variations in the vitelline glands and ducts of Opisthorchis pseudofelineus

Ward. Only those groups of the vitellaria which are near the ovary are

shown, the underlined figures indicating the number of the group, counting

caudad in the antovarial region and cephalad in the postovarial region.

Fig. 1. “Y” type of vitelline ducts, ventral view. X 14.

Fig. 2. “‘V” type of vitelline ducts, ventral view. X 21.

Fig. 3. Variation in position of break between the regions of the vitel-

larium, ventral view. X 14.

Fig. 4. “V”-“Y” type of vitelline ducts, dorsal view. X 14.

Fig. 5. Median group of vitellarium, opposite ovary, dorsal view; 2, 3,

5, 6, number of vitelline glands. X 14.

Fig. 6. Accessory vitelline duct, dorsal view. X 14.

Fig. 7. Regions of vitellarium connected by duct, ventral view.

Fig. 8. Median group of vitellarium opposite ovary, dorsal view.

Fig. 9. Regions of vitellarium connected, dorsal view. X 14.

Fig. 10. Accessory vitelline duct, dorsal view. X 14.

Fig. 11. Single vitelline duct on left side, ventral view. X 14.

Fig. 12. “T” type of vitelline duct, dorsal view. X 14.

Fig. 13. “Y” type of duct, drosal view. X 14.

Fig. 14. “V” type of duct, dorsal view. X 14.

Fig. 15. “V” type of duct, dorsal view. X 14.

x X

RE

I10 F. D. BARKER

Plate IX

Fig. 1. Opisthorchis lancea Dies., ventral view. After Weski, 1900:

582. X Io.

Fig. 2. Opisthorchis pseudofelineus Ward, dorsal view, showing degen-

erate vitelline glands. X II.

Fig. 3. Opisthorchis felineus Riv., ventral view. X II.

Fig. 4. Optsthorchis peluH drole Ward, dorsal view, showing degen-

erate vitelline glands. X II.

Fig. 5. Opisthorchis pseudofelineus Ward, ventral view. X 11.

Fig. 6. Opisthorchis pseudofelineus Ward, acetabulum and genital pore.

X 30.

PUATESIX

F D.Barxer del

SOME STUDIES ON TRYPANOSOMA LEWISI

By LEROY D. SWINGLE

WITH ONE PLATE

During the last four years the literature on trypanosomes has

increased marvelously. New species are continually being added to

the list, while the morphology and life history of the older forms are

being now worked out. Schaudinn’s remarkable memoir (1904) on

the life cycle of Trypanomorpha noctuae and Trypanosoma ste-

mann has paved the way for the discovery of the life histories of

the other forms, and since his work appeared, Prowazek (1905) has

discovered that the rat trypanosome passes through stages in the

rat louse similar to those found by Schaudinn in the mosquito. It

is with the rat trypanosome that the present paper deals.

Woodcock (1906), who has admirably reviewed the present know!l-

edge of haemoflagellates, substitutes the term trophonucleus for

nucleus and kinetonucleus for blepharoplast, since the former has

to do with nutrition and reproduction, and the latter with locomo-

tion. The fact that the locomotor apparatus develops from the

kinetonucleus, as Schaudinn has shown, would seem to justify

Woodcock’s terminology. For reasons too numerous to give here,

he also changes the orientation of the trypanosomes, designating the

end formerly considered anterior, as posterior, and vice versa. In

this discussion I follow Woodcock’s terminology, but for reasons

which will be given later do not accept his orientation.

The trypanosome of the rat has a geographical distribution coex-

tensive with that of its host. To the recorded places where rats are

known to be infected I can also add Seattle, Washington. But, as

has already been noted by other authors, the infection in a given

city is localized. In Lincoln, Nebr., rats caught at two places a

square apart showed infection, while none was found in those from

other parts of the city. At one of these centers thirty-six per cent.

I12 LEROY D. SWINGLE

of the rats were infected. Adults seemed to be exempt from the

disease, but fifty-three per cent. of the rats about half grown and

twelve per cent. of those about one-fourth grown were infected.

- Twenty-three per cent. of all the rats caught were harboring the

parasite.

Two field mice and six house mice were examined for trypano-

somes, but with negative results. However, the culture method for

the detection of the parasite was not used.

A two-thirds grown rat, heavily infected, became sick and died.

Several hours afterwards an examination of various parts was made.

The parasites were very numerous in the liver, the veins of the fore-

limb, and the blood sinuses of the brain. They were less abundant

in the spleen, the kidneys, the lungs, and a lymph gland from

the axilla. The parasites possessed great activity in spite of the

fact that the rat had become very cold and stiff. No hypertrophy of

the organs could.be detected. The parasites could be found easily

in sections of the tissue by staining with carbol thionin. No intra-

cellular stages were discovered.

With regard to technique, iron haematoxylin and eosin give fair

results. They bring out the trophonucleus, kinetonucleus, and flagel-

lum plainly. But the trophonucleus takes the stain too readily for

the cytoplasm. If the specimen is destained enough to differentiate

the chromatin of the trophonucleus, the cytoplasm is not sufficiently

stained. Goldhorn’s stain was used with mediocre results. But the

best differentiation was obtained by the use of Wright’s stain. This

stain, however, is quite variable, due to the evaporation of the alco-

hol. For the best results the alcohol must be sufficiently saturated

with the stain to give a rich reddish-purple when water is added.

In this condition, if the stain be left on the smears from ten to

fifteen minutes, the flagellum will take on a deep red color, the cyto-

plasm a light blue, the trophonucleus a rose-red and the kineto-

nucleus a darker red. When the parts are thus stained, the connec-

tion between the flagellum and kinetonucleus is plainly shown, the

flagellum staining right up to the latter.

Authors are very well agreed in regard to the morphology of

the adult trypanosome. MacNeal (1904), however, noted that some

of the commonly called adults were enlarged and possessed two tro-

phonuclei as late as the one hundred and tenth day of the infection.

He explains this as a slow and retarded longitudinal division. Sev-

STUDIES ON TRYPANOSOMA LEWISI 113

eral days after the period of multiplication had passed, I found some

parasites with two kinetonuclei and others with two trophonuclei

(pl. x, fig. 1). This is not a result of mechanical division of the

trophonucleus by smearing, but must be regarded either as a longi-

tudinal division or as an abnormality.

There is considerable disagreement among workers about the

mode of multiplication of the trypanosome in the blood of the rat.

Rabinowitsch and Kempner (1899) have described three types:

longitudinal division, transverse division, and segmentation in which

the undulating membrane and flagellum disappear. In their later

work (1903) they modified their views, but still held that the forma-

tion of rosettes in which the mother cell could not be found does

occur. Francis (1903) described and figured the same modes of

division as Rabinowitsch and Kempner in their first work. But he

says: “ These three forms of division are separated only for descrip-

tion. They may all be seen side by side in the same preparation

and in some instances the distinction between them is not altogether

clear, especially between the longitudinal and transverse modes.

Division by segmentation begins with a curving of the parasite

which continues until the two ends approach each other and finally

meet, thus giving the parasite a globular form. 'The flagellum be-

comes lost and the undulating membrane disappears.” Laveran and

Mesnil (1904) recognized two types of division which they desig-

nated as group aand group b. In the first group the mother trypan-

osome can be recognized, being larger than the other individuals,

while in the second group the individuals are all the same size.

Danilewsky distinguished two modes of division, namely, longitu-

dinal division and segmentation. In the latter he says that the

flagellum and undulating membrane disappear. MacNeal’s obser-

vations (1904) agree with those of Wasielewski and Senn (1900),

who maintain that unequal longitudinal division is the only method

of multiplication.

My observations agree most nearly with those of Laveran and

Mesnil. In both living and stained preparations I found each kind

of rosette. By an unequal longitudinal division of the mother try-

panosome a small cell is formed (pl. x, fig. 3). Several such

divisions give rise to a rosette consisting of a number of small, equal

cells and the large mother trypanosome (pl. x, fig. 4). Further,

each one of the small individuals may divide by equal longitudinal

114 LEROY D. SWINGLE

division before separating. In fact small cells divide regularly by

this method, and couplets thus produced are found in the blood again

and again. And so, when such division proceeds without a com-

plete separation of the individuals, a rosette lacking a large mother

trypanosome is formed. A similar rosette would be formed if the

mother trypanosome of group a should become broken off. Thus,

rosettes lacking the mother trypanosome belong to group b and are >

derived from group a, as Laveran and Mesnil have already pointed

out. A study of rosettes in living preparations gives quite a differ-

ent idea of their formation from that gained by the study of smears.

The individuals do not lie in a single plane, as they are seen ina

smear (pl. x, fig. 2), but lie parallel to each other with their

flagella all directed the same way (pl. x, fig. 4), unless division has

more or less completely taken place. Such rosettes would probably

be crushed in spreading and thus give rise to the many fantastic

shapes which one always finds in smears. However, if division has

proceeded till the individuals are free except at their posterior tips,

then in spreading a more or less perfect rosette would be formed

(pl x,t. 2)\

But there is another divisional form which, as far as I am aware,

has not been described by any previous author. A one-fourth

grown rat was inoculated subcutaneously February 26 with trypa-

nosomes. On March 3 multiplication forms, both large and small,

appeared simultaneously. On March 5 the blood showed a heavy

bacterial infection which increased till March 7, when the blood

presented a very light pink or almost milky color. Not only the

tail blood had this appearance, but also the blood taken from the

feet. It began to clear up on March 8 and the following day re-

sumed its normal color. The rat died March 11 without showing other

symptoms than sleepiness. Examination of the fresh blood revealed

peculiar forms, some of which looked as if there was a flagellum at

each end, the motion being quite different from the usual character.

The true structure of these peculiar forms was not evident, until

stained preparations were studied, when it was found that there

were two types present. The individuals belonging to the first type

are somewhat smaller than an adult trypanosome (pl. x, fig. 5);

those belonging to the second type larger than an adult (fig. 6).

The forms belonging to these types were quite numerous, not fewer

than thirty of each kind having been found in a few smears. They —

Nee eee 2

STUDIES ON TRYPANOSOMA LEWISI I15

could be picked out very easily, being so similar to each other and

so different from the ordinary divisional forms which existed side

by side with them (compare pl. x, figs. 6, 7, 8, Io and 11, also

figs. 5 and 9g). Although these represent various stages of division,

yet it is evident that they all belong respectively to the given types.

The interpretation that should be put on these forms seems doubt-

ful. Evidently the intense bacterial infection was a factor in the

cause for their existence, since they did not occur before the appear-

ance of the bacteria. It might be conceived that under this adverse

condition to which they were subjected they took to conjugation.

But since Prowazek’s work on the trypanosome in the rat louse,

this interpretation would hardly seem reasonable. The explanation

which appears most satisfactory is that they are cases of equal longi-

tudinal division, where the flagellum and undulating membrane split

their entire length. MacNeal (1904) contends that the flagellum

does not divide, but that the new flagellum grows out of the new

kinetonucleus in such close proximity to the old one that it appears

to split off from it. Laveran and Mesnil (1904) say that the new

flagellum is formed by the division of the old one from the kineto-

nucleus out to the undulating membrane, at which point it breaks

out and then lengthens by growth. At any rate, the new flagellum

in ordinary cases can be distinguished by the fact that it is shorter

and thinner than the old one. But in these cases the flagella are

equal in length and thickness, and neither a mother flagellum nor

even a mother trypanosome can be distinguished. That the flagel-

lum does actually split is substantiated by many forms (pl. x, figs.

10, 11). In some (fig. 10) the flagellum has split from the kineto-

nucleus to a point anterior to the trophonucleus, and in others (fig.

II) nearly to the trophonucleus. After the flagellum has split its

entire length one can easily see how a bilaterally symmetrical form

would result in spreading (figs. 6, 7 and 8). In those cases (fig.

5) where the division of the flagellum seems to have been from the

anterior to the posterior, it will be noticed that the kinetonucleus is

tardy in division, and we would therefore expect the flagellum to

divide in this way, the trophonucleus and anterior cytoplasm having

divided before the kinetonucleus. Whatever may be the true expla-

nation of these forms, the fact remains that they are entirely dis-

tinct from any form which occurred in the other rats which were

inoculated.

116 LEROY D. SWINGLE

Byloff (1904) described and figured a kind of mitosis. Many of

my preparations show: forms quite similar to his figures, but they

occur most abundantly toward the close of the multiplication period

and are generally very vacuolate. Moreover, there is not enough

definiteness about the arrangement of the chromatin to lead me to

regard them as mitotic figures. |

In several rats parasites in actual process of division were very

scarce in the peripheral circulation, and yet the number of trypano-

somes in the blood was increasing rapidly. To be sure, many en-

larged forms which were preparing for division and a very few

small rosettes were found. Two of the rats were killed and exam-

ined. In one rosettes were found to be quite abundant in the spleen ;

in the other the bone marrow was teeming with them, amply suffi-

cient in number to account for the rapid increase of the parasites

in the peripheral blood.

Agglomeration was secured under the following conditions: A

one-fourth grown rat was inoculated intraperitoneally with a few

drops of blood bearing a rather light infection. Twenty hours iater

the trypanosomes were found in the blood from the tail and resem-

bled those inoculated. The next day agglomeration began and

twenty-four hours later rosettes consisting of as many as sixteen

sndividuals were found. A few days later the parasites had entirely

disappeared. No divisional forms were discovered, but, judging

from the small number inoculated and the comparatively large

number agglomerated, a multiplication must have taken place some-

where in the body. The agglomeration presented some unique

forms. Two of the parasites would unite by the posterior and

anterior ends, respectively, so that as they moved through the field

they gave the appearance of a loop increasing and diminishing with

the change in motion of the bodies. Such couples were quite numer-

ous, and were never seen to separate from each other at either end

(pl. x, fig. 12). |

As MacNeal has observed, the contact of individuals is not merely

by their posterior tips, but they overlap. He says that in Trypa-

nosoma brucei a perfect fusion takes place, but does not make it

clear whether that is the case with Trypanosoma lewist. At any

rate, in some cases of the latter a complete fusion takes place and

the kinetonuclei come to lie close together or apparently fuse (pl.

x, fig. 12). Sometimes, when several are agglomerated, the out-

Se ee

2 eas Ee

STUDIES ON TRYPANOSOMA LEWISI 17

lines of the posterior ends are entirely lost and the kinetonuclei all

lie close together in a central mass.

As to the mode of transmission, natural infection is doubtless

accomplished by the bites of blood-sucking insects, such as lice and

fleas. Rabinowitsch and Kempner (1899) succeeded in transmitting

the disease by placing on a fresh rat fleas which were taken from an

infected one. Similarly, MacNeal (1904) obtained a positive infec-

tion by the use of lice. But up to the year 1905 none of the inves-

tigators had found other than adult stages in infected lice and fleas.

In 1905, however, Prowazek published an account of the life cycle

of the parasite observed in the louse. My observations show that it

probably passes through a similar cycle in the flea.

Examination of fleas teased in normal salt solution imme-

diately after their removal from an infected rat showed many small

oval trypanosomes which had lost their flagella, spindle-shaped

forms, and large active ones resembling the adults in the rat.

Several with long flagella and with the posterior half of the body

bent forward on the anterior part were found. In a flea examined

twenty-four hours after it was taken from an infected rat rosettes

of four and eight trypanosomes respectively were found. The cells

were somewhat thickened and from their motion it was evident that

the flagella were centrally located. In another flea two instances

were found where agglomeration by the posterior ends had taken

place and been followed by a rounding off of the ends, the flagella

still persisting. Rosettes of at least seventy-five individuals with

their flagella centrally located were found in another flea at the

same time.

By sectioning and staining with iron haematoxylin the viscera of

the infected fleas, the parasites and their relation to the host could

be made out. In the case of a flea which was sectioned forty hours

after its removal from the rat very few trypanosomes were found

in the stomach, but great masses lined the wall of the rectum. Be-

sides the large masses attached to the wall, here and there small

groups were found attached by their flagella (pl. x, fig. 13). Such

ones were also found attached to the epithelial cells of the stomach.

Rosettes in which the flagella were directed centrally were found

free in the rectum (pl. x, fig. 14). Between the epithelial cells of

the stomach and in the intestine masses of trypanosomes were found

in which the flagella had entirely disappeared and the cells become

spherical (pl. x, fig. 15).

IIS LEROY D. SWINGLE

In this connection I wish to call attention to two of the reasons

why Woodcock regards the non-flagellate end as the anterior. His

argument is as follows: When herpetomonadine forms, such as

Trypanomorpha, enter a resting phase, they attach by the flagellate

(anterior) end. Also the heteromastigine form, Trypanophis, at-

taches by the anterior end. But, when Trypanosoma ziemannt

enters a resting phase, it attaches by the non-flagellate end. More-

over, there are certain other species of trypanosomes which are

known to attach to blood corpuscles by the non-flagellate end. Now,

since forms in which the orientation is known attach by their ante-

rior ends, he believes that all trypanosomes attach by the anterior

end, and since in all cases where the trypanosome is known to attach,

it attaches by the non-flagellate end, he would regard this end as

the anterior. The second reason is based on the mode of agglom-

eration. In Trypanomorpha, agglomeration takes place by the fla-

gellate (anterior) end. In sharp contrast to this the trypanosomes

agglomerate by the non-flagellate end. This end must therefore be

the sensitive pole. Since forms in which the orientation is known

agglomerate by the anterior end, and since the end by which agglom-

eration takes place must be considered the sensitive pole, he regards

the non-flagellate end of the trypanosome as the anterior.

My observations, however, show that Trypanosoma lewnsi at-

taches by the flagellate end (pl. x, fig. 13) and sometimes forms

rosettes with the flagella centrally located. Perhaps these rosettes

are not agglomerations, but nevertheless the ends directed centrally

should be regarded as the sensitive poles as much in this case as in

cases of agglomeration. Woodcock gives other reasons which may

still justify him in considering the non-flagellate end of trypano-

somes as the anterior, but these exceptions would seem to invali-

date in a measure his generalizations.

The rosettes found in the flea may be merely cultural forms sim-.

ilar to those found by Novy, MacNeal, and Torry (1906) in the

stomachs of mosquitoes fed on infected blood. But in view of

Schaudinn’s work on the owl trypanosome in the mosquito and

Prowazek’s work on the life cycle of the rat trypanosome in the

louse, it is very probable that other than mere cultural rosettes are

to be found in the flea. The discovery of attached forms as already

described in this paper seems to confirm this opinion and to demon-

strate that the flea also acts as an intermediate host.

STUDIES ON TRYPANOSOMA LEWISI 119g

A bit of circumstantial evidence to the effect that the flea plays

an important role in the transmission of the parasite is worthy of

mention here. Of the seventeen one-fourth grown rats examined

in the autumn and winter not one was infected with either lice, fleas,

or trypanosomes, while of the seven caught the following spring at

the same place all had fleas and four were harboring the parasite.

This was the only place in the city where infection was found to

exist, and, moreover, neither lice nor fleas were found on those rats

caught in other parts of the city. From this evidence and also from

the fact that fleas were found on infected rats much more often than

lice, one would conclude that the flea served as the intermediate

host more often than the louse. While the life cycle of the parasite

in the former is doubtless quite similar to that in the latter, yet an

excellent field for further investigation is presented here.

This work was done in the Zoological Department of the Univer-

sity of Nebraska under the direction of Dr. Henry B. Ward, to

whom I am indebted for interest and assistance in the investigation.

LITERATURE CITED*

By Lorr, K.

1904. Ein Beitrag zur Kenntniss der Rattentrypanosomen. Sitzungsb.

k. Akad. Wissensch. Math.-naturw. Kl, Wien, (Abt. III.) CXIII:

III-138.

Francis, E.

1903. An experimental Investigation of Trypanosoma Lewisi. Bull. Hyg.

Laby., No. 11: I-26.

LAVERAN, A., AND MEsSNIL, F.

1904. Trypanosomes et Trypanosomiases. Masson et Cie.; Paris. (8°,

418 pp., I pl., 61 figs.)

MacNEAt, W. J.

1904. The life-history of Trypanosoma Lewisi and Trypanosoma Brucet.

Jour. Infect. Diseases, I: 517-543.

Novy, F. G., MAcNEAL, W. J., AND Torrey, H. N.

1906. Mosquito Trypanosomes. Science, N. S., XXIII: 207-208.

PROWAZEK, S.

1905. Studien iiber Saugetiertrypanosomen. Arb. kaiserl. Gesundheits-

amte, XXII: 351-395.

RaBINOwITSCcH, L., AND KEMPNER, W.

1899. Beitrag zur Kenntniss der Blutparasiten, speciell der Rattentry-

panosomen. Zeitschr. f. Hyg., XXX: 251-204.

Since Laveran and Mesnil (1904) have monographed the trypanosomes,

and Woodcock (1906) has given an extended bibliography in connection with

his review, there is need to cite only the papers directly bearing on this article.

120 LEROY D. SWINGLE

1903. Die Trypanosomen in der Menschen- und Tierpathologie, sowie

vergleichende Trypanosomenuntersuchungen. Centralbl. f. Bakt.,

XXXIV: 804-822,

ScHAUDINN, F.

1904. Generations- und Wirtswechsel bei Trypanosoma und Spirochaete

(Vorl. Mitteilung). Arb. kaiserl. Gesundheitsamte, XX: 387-438.

WASIELEWSKI, V., AND SENN, G.

1900. Beitrage zur Kenntniss der Flagellaten des Rattenblutes. Zeitschr.

f. Hyg., XXXIII: 444-472.

Woopncock, H. M.

1906. The Haemoflagellates: a Review of Present Knowledge relating to

the Trypanosomes and allied forms. Quart. Jour. Micr. Sci, L:

151-331.

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STUDIES ON TRYPANOSOMA LEWISI I21

EXPLANATION OF PLATE

Plate X

All figures are magnified approximately 1500 diameters.

Fic. 1. Adult in process of division several days after multiplication

period.

Fig. 2. Multiplication rosette showing mother cell and several daughter

cells.

Fic. 3. Early stage of unequal longitudinal division as seen in living prep-

arations.

Fig. 4. A later stage than Fig. 3. The cells lie parallel to each other, the

anterior ends in the same direction.

Fic. 5. Equal longitudinal division of a form smaller than an adult. The

flagellum has split from the anterior to the posterior before the kinetonucleus

has divided.

Fic. 6. Equal longitudinal division of a form larger than an adult. It

presents bilateral symmetry.

Fics. 7, 8. Forms corresponding to Fig. 6. In the first, the kinetonucleus

has divided twice.

Fic. 9. An earlier stage than Fig. 5. Cytoplasm and trophonucleus not

yet divided.

Fics. 10, 11. Earlier stages than Fig. 6. Flagella in process of division.

Fic. 12. Agglomeration of two individuals by anterior and posterior ends

respectively.

Fic. 13. Three trypanosomes attached by their flagella to the wall of the

rectum of the flea.

Fic. 14. Rosette free in the rectum. The kinetonuclei lie anterior to the

trophonuclei; the flagella are directed centrally.

Fic. 15. Mass of trypanosomes found free in the intestine. The indi-

viduals have become spherical and lost their flagella, but retain their kineto-

nuclei.

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NEW FORMS OF VOLVOX

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WITH FOUR PLATES

(Studies from the Zoological Laboratory, the University of Nebraska, under

the direction of Henry B. Ward, No. 75.)

The typical genera of the Volvocineae, Pandorina, Eudorina, and

V olvox, are cosmopolitan and well known types. It has even been

suggested that Volvox at least is so cosmopolitan that new species

are not likely to be met with. Yet within a few years the writer

has found, in one vicinity and without especial search, new forms,

or at least decidedly new phenomena, in connection with each one

of the above genera. Pandorina has shown a wholly unwarranted

method of asexual reproduction, a method in which single cells,

instead of all of the cells, gave rise to new colonies, suggesting, at

least, the parthenogenesis of Volvox. LEudorina has shown all pos-

sible transition gradations between its proper self and the newly

discovered form—supposed to be a separate genus—of Pleodorina,

the four “ somatic cells ” of the latter proving somatic or not somatic

according to the degree of their differentiation in size.

Lastly, Volvox, upon the two occasions of its occurrence within

the writer’s observation, has differed so fundamentally from hitherto

described forms as to suggest the necessity of specific separation.

Even in the two instances in which it has occurred, in ponds but a

few miles from each other, the forms were not only new, but they

differed from each other in certain important respects as much as

all previously described forms have differed in regard to the same

characters. Unfortunately, in the case of both of these finds, it was

impossible to follow the species for any length of time to note con-

stancy or inconstancy throughout changing seasonal conditions.

One pond was dry within a week after the Volvox was noted.

The other was inconvenient of access, and the organism was not

*I owe the first demonstration of this fact to my former pupil and assis-

tant, Mr. George R. LaRue of Crete, Nebraska.

124 J. H. POWERS

found until late in the season. For these reasons I have delayed

describing the forms, hoping to hit upon more material and work

out more fully the relation of the types to the known forms of the

genus. But our seasons are irregular and Volvox, everywhere pre-

carious, seems but an occasional visitor with us. A description of

the forms as found may draw attention to the possibilities that await

a fuller search elsewhere. Moreover, the forms are not uninterest-

ing taken by themselves. I will describe the two forms separately,

comparing each with the forms of the genus hitherto recorded, viz.,

Volvox globator; V. aureus (==V. minor), with its important

variety described by Klein; V7. cartert, the doubtful species de-

scribed from India; and lastly V. tertius, discovered and described

by Arthur Meyer? in Germany in 1806.

The material upon which the description of the first form is based

consists at present of thirty mounted slides, containing several thou-

sand colonies and displaying in abundance all phases of sexual and

asexual reproduction. The collection was made several years ago,

at about midsummer, in the shallow remnant of a prairie pond. It

was without drainage and contained considerable alkali. The pond

was exceedingly variable, covering several acres during a good part

of some seasons, while remaining nearly or quite dry throughout

others. It is a favorite resort of water birds during their migra-

tions, and the Volvox may thus easily have been introduced from a

distance. The material, after considerable examination in the fresh

state, was fixed successfully in a number of reagents, the most per-

fect results being obtained with Flemming’s strong formula diluted

with several volumes of water. Stains worked easily and well, and

mounts were made both in glycerine and balsam, chiefly in the

former.

Of a number of features in which the Volvox in question differs

from the members of the genus already described, the most interest-

ing is the manner of formation of the sperm cells. These elements

arise in all the above mentioned forms by the rapid division of

enlarged cells of a parent coenobium. Sperm platelets are thus —

formed, containing variable numbers of sperms, and lying, until

Ludwig Klein: Neue Beitrage zur Kenntniss der Gattung Volvox,

Berichte d. deutschen botanischen Gesellschaft, VII: 42.

? Arthur Meyer: Die Plasmaverbindungen und die Membranen von Vol-

vox globator, aureus, und tertius, mit Riicksicht auf die thierische Zellen,

Botanische Zeitung, XI and XII. 1806.

NEW FORMS OF VOLVOX 125

their final extrusion, within the common wall of the parent colony.

The number of such sperm platelets may vary within wide limits.

In V. globator and V. carteri it remains but few. In V. aureus and

V. tertius it may become very great, until in rare cases of so-called

“ Sphaerosira colonies” of V. aureus two-thirds of the original num-

ber of the cells of the colony (1,000 to 1,100 in all, according to

Klein) may be transformed into sperm mother cells and so ulti-

mately into sperms. It should be noted, however, that even in such

cases the typical character of the colony is preserved. It retains its

free locomotion and especially its polarity. The sperm bundles are

most numerous at the posterior pole, diminishing toward the anterior

pole, which always remains free from them. The sperms, too, do

not develop simultaneously ; those at the posterior pole are developed

and discharged first.

In the form which I am to describe, the phenomena are so differ-

ent as, at first sight, to appear unrelated. Not a single instance

could be found of a sperm platelet developing at the periphery of

a free-swimming, mature colony. I examined many hundreds, if

- not thousands, of the living coenobia, and have also examined every

coenobium of suitable development among my mounted prepara-

tions, probably not less than a thousand in all. The great majority

are in sexual reproduction, yet not an instance of typical sperm pro-

duction occurs. Instead I meet everywhere, within the older coeno-

bia, with daughter colonies, or with what look like daughter colonies,

every cell of which gives rise to a sperm platelet. (Pls. x11 and XIII,

figs. 7, II, 12, 14, 15, 16, 17.) 1 am obliged to choose a new term

to designate these premature, exclusively reproductive, and exclu-

sively male colonies, if such they truly are. The name “ sperm

colony” would be most natural, had it not been applied by several

writers to the single sperm platelet; while the term “ male colony ”

has been applied to developed and usually independent coenobia

whose reproductive cells become sperms only, instead of eggs or

parthenogonidia. I choose for this purpose the designation “ sperm

spheres,” or the phrase “ pure male colony.” As already suggested,

I find this method of sperm production invariable. All in all I have

certainly examined hundreds of sperm spheres in their mature con-

dition. e., made up of platelets of ciliated sperms,—and of the

more or less immature stages I have examined many hundreds more ;

yet among them all I have been unable to find a single ‘“ somatic

126 J. H. POWERS

cell” ;* every cell, or every “ individual” of the young colony, in-

variably becomes a sperm mother cell; none remain as ova; each

becomes a platelet of sperms. The entire uniformity of this method

is, if possible, even more significant than its novelty. And it is this

fact which, under any interpretation which may be put upon it, sepa-

rates this form of Volvox sharply from the others which most nearly

approach it.

T'wo very different interpretations may, in fact, be placed upon

this new instance of sperm development. We may, on the one hand,

consider it an instance of acceleration and, in a sense, hypertrophy :

acceleration, because the sperms are matured in very young colonies

which pass all their life within the parent coenobium; hypertrophy,

because, instead of a few or even the majority of the cells becoming

sperm bundles, all of them become sperm bundles. On the other

hand we may, in a measure, reverse our mode of interpretation and

consider these sperm spheres, not as instances of acceleration, but

of retardation in sperm production; they become, if thus viewed, not

true colonies at all, but instances of the multiplication of primary

reproductive cells—‘“ spermogonia”’—comparable to that which

takes place in the metazoon testis. As we shall see later there are

facts which point toward both of these interpretations, and perhaps

the majority point toward the latter. Doubtless these two hypothe-

ses are not, in truth, as different as they really seem—not, that is,

for an organism on the developmental plane of Volvox. None the

less, they are not one and the same, and from the standpoint of our

definitions they make a great difference; the one throws this Volvox

back closer to the Protozoa and relates it far more closely to Eudor-

ina, the other makes it quite violently metazoon. For the present

let us assume, as the least radical, the first of these hypotheses, z7z.,

that these sperm spheres are young colonies disintegrating wholly —

*I shall deliberately use, in this paper, such expressions as “somatic

cells,” “reproductive cells,” etc., and I shall also use, with equal freedom,

the terms “colony,” “coenobium” and the like. Such expressions are flatly

contradictory, in a sense; but so are the facts. They confuse no one familiar

with the different points of view from which Volvox may be considered. The

very beauty of Volvox and its group lies in the happy way in which they over-

ride “fundamental distinctions.” Out of the seeming chaos, therefore, of

terms—terms old and terms new, terms botanical and terms zoological, terms

metazoon and terms protozoon,—I choose those most convenient and useful

for the context in question.

NEW FORMS OF VOLVOX 127

into sperms, and make comparison with the known forms which

most nearly approach it.

These forms are Volvox tertius as described by Meyer on the one

hand, and, more particularly, the extreme form of V. aureus as

described by Klein on the other. The facts presented by these types

are very important for the understanding of the form I have dis-

covered, for, if the hypothesis we are assuming be correct, they cer-

tainly constitute connecting links between it and the more typical

forms of the genus. A brief résumé of the facts recorded by Klein

will suffice equally well for both forms, the two being much alike

in the matter of sperm production and Klein’s description being

much the more adequate. Probably, too, the variation which he

found in V. aureus was more extreme, and resembled more the form

found by the author than did V’. tertius found by Meyer.

After spending much time in the study of the genus, and espe-

cially in investigating the variation of the species V. aureus in

diverse localities and under diverse conditions, Klein published an

extended monograph! upon the subject. In this he recorded a con-

siderably wider range of variation for V. aureus than had hitherto

been known. He showed conclusively, what had been already sus-

pected, that the species was not necessarily, though it was typically,

dioecious, and he increased considerably the number of sperm plate-

lets which might be found in a single colony. The results of his

extended research did not, however, go farther than this. After

the close of his own investigations, however, he received a few pre-

pared slides from a friend in Marburg, which showed a form of V.

aureus far more suggestive of the Nebraska type. From his de-

scription and figures we gather that it was a small representative of

the species (approximately 250 to 350 p), this fact being due rather

to a meagre development of the gelatinous matrix than to a reduc-

tion in the number of cells, or the size of the cells, in the colonies.

The surprising fact, however, was the great precocity of the develop-

' ment of the daughter colonies while within the parent coenobia.

They showed the ordinary cell-numbers of the species V. aureus, but

they became of rather large size for young colonies, and showed a

surprising acceleration in the development of reproductive cells.

*Ludwig Klein: Morphologische und biologische Studien uber die Gat-

tung Volvox. Pringscheim’s Jahrbiicher fiir wissenshaftliche Botanik, XX:

133.

128 J. H. POWERS

Ova reached a considerable size within the young coenobia, a size

much beyond the ordinary, while the sperm platelets were frequently

constituted of completely developed, ciliated sperms. These sperms

and the platelets were a little smaller than is typical for the species

V. aureus, but not smaller than is sometimes found. The only

noticeable peculiarity was the invariable number of sperms in a

platelet,—thirty-two,—instead of the usually variable assortment of

large and small bundles.

The apparent resemblance of this interesting variation (which in

these respects runs nearly parallel to the new species V. tertius) to

the form found by the author is plain. In each we have an absence

of sperm platelets in the original mother (grandmother?) coenobium.

In each the sperm platelets, or at least a part of them, are formed

and matured in daughter colonies, while within the original coeno-

bium. In each we find, if we count the sperm bundle as the homo-

logue of a colony, in the words of Klein, “drei in einander einge-

schachtelte Generationen; von denen jede vollkommen ausgebildet

ist.” In each we have the “ grauelvolles Familienbild” presented

by the possibility that, within the body of the grandmother volvox,

sperms should fertilize ova which stand to them in the relation of

grand aunts. For ova are present in the older coenobia side by side

with the precocious daughter colonies.

But a closer comparison shows at once that the points of differ-

ence are as many and even more significant than are the points of

similarity.

In Klein’s V. aureus colonies there was an irregular but general

acceleration in the development of all reproductive cells, sperms,

ova, and, presumably, parthenogonidia. In the form I am describ-

ing there is an actual and general retardation in the development of

parthenogonidia greater than seems to take place in any other de-

scribed Volvox, and there is at least no acceleration of the ova. I.

can never distinguish between ova and parthenogonidia in the undis-

charged daughter colonies. The development of sperms only is

hastened, if such be the true interpretation, and, as we shall see, they

are discharged from the “individuals” that produce them before

these “ individuals,’—the sperm spheres,—are born. Here lies a

difference that separates the two cases widely. The daughter col-

onies described by Klein, although not observed by him in the living

state, were, he assumed, still capable of an independent existence

and were destined to escape from their parent coenobia and lead

NEW FORMS OF VOLVOX 129

such an existence, even though some of their mature sperm platelets

should have become free and functional before their birth. I paid

careful attention to this point when examining the living material.

But an extended search did not discover a single sperm sphere in

the free-swimming condition. On the other hand I did find sperm

spheres which were breaking up into theit component sperm bundles

while still within the parent coenobium. The disruption begins by

a more or less general loss of form and shifting of position of the

individual bundles due to the action of their cilia, after which they

escape, singly or a few at a time. My permanent preparations show

an abundance of such stages. (PI. xuu, fig. 17.) I regret that I

did not make the experiment of freeing these sperm spheres artifi-

cially, to see whether or not they were capable of locomotion as a

whole. In any case such motion is not natural to them, although

the spheres, just before the final cell divisions, do show cilia, or at

least rudiments of them. (Pl. xu1u1, fig. 14. The cilia could be

made out with difficulty and do not show in the figure.) Correlated

with the entire loss of somatic cells and of the power of independent

existence goes the loss, or absence, of another otherwise universal

character of Volvox colonies, vig., their polarity. Even Klein’s pre-

maturely developed daughter colonies, which, as he says, certainly

passed an unwonted amount of their existence within the parent

coenobium, yet showed distinctly the polar differentiation. Many

of them are figured as ellipsoid, and they evidently all retain the

characteristic area of exclusively somatic cells at one end. Whether

or not the sperms ripened, as usual, in succession, from the more

reproductive toward the somatic pole, his figures do not show, but

had it been otherwise he probably would have mentioned the fact.

As above suggested, no trace of this polarity seems to be retained

by the sperm spheres which I am describing. The sister coenobia,

which often develop side by side with them in the same parent, show

such polarity strongly. (PI. x1, figs. 1 and 3, and pl. xu, figs. 7, 8

and 9.) They are usually strongly ellipsoidal and their reproduc-

tive anlagen are always confined to one-half or two-thirds of the

ellipsoidal area. The pure male colonies, on the contrary, are

characteristically spherical in form even from the first, unless they

are flattened against the wall of the parent colony. (Pl. x1, figs. 2

and 3; pl. xu, figs. 7, 8, 11 and 12; and pl. x11, figs. 14, 15 and 16.)

They lose this form only when approaching disruption. (Pl. xz1,

fig. 17.) Not only are they without the somatic area, being without

130 J. H. POWERS

all somatic cells, but the sperm platelets and the preceding genera-

tions of cells which produce them all develop with great uniformity,

and all become finally mature at the same time. (PI. xu, fig. 12;

pl. xi, figs. 14, 16, 17.) The only exception is the temporary

irregularity in the sizes of the cells during some of the last divisions

that form the sperms (pl. x11, fig. 15) ; but this irregularity is tem-

porary only, and has nothing to do with the perfect equality of all

sides of the sperm sphere as such.

From all of these facts it becomes plain that the sperm formation

of the Volvox we are describing is very different from even the

types which most closely resemble it. If these sperm spheres are

phylogenetically mere parthenogenetic daughter colonies it is plain

that they are daughter colonies that have become so subordinated to

the single function of sperm production that they stand to the parent

organism upon the functional plane of a mere spermary. Only the

tangled web of homologies prevents our so recognizing them at —

once. The facts of development will throw a little more light upon

the question. To them we shall shortly return, but first a fuller —

description of the sperm spheres.

The number of sperm platelets in the sperm spheres follows very ©

closely if not exactly the theoretical numbers 64, 128, and 256, there ©

being these three sizes and these only. I have made many counts

which give me sometimes the exact numbers, although it is difficult —

to avoid a certain range of error which is of course greatest with ©

the highest numbers. These results may be checked somewhat by

counting the numbers in the hemispheres in plate x11, figures 14, .

15, and 16. Plate x11, figure 12, may also be used. Figure 14

represents one-half of a large-sized sperm sphere in the spermo- |

gonia stage and will be found to show very close to 128 cells; figure

12 is a mature sphere of corresponding size and shows a similar num-_

ber ; figures 15 and 16 are of intermediate colonies and give approxi- |

mately one-half the number of individual groups. Near the center of -

figure 16 a blurred space may, be seen large enough to contain two”

sperm bundles. Under the microscope it did show these bundles, but

they were disintegrating and plainly degenerate cells. It is probable |

that a few cells may degenerate at any stage and so reduce, in some

cases, more or less the final number of sperm platelets. My count indi-

cate this. Nevertheless the number seems usually regular. As to.

the frequency of these three sizes of sperm spheres, the smallest

ee

NEW FORMS OF VOLVOX 131

are much the most infrequent, and while the largest are frequent,

by far the greater number are of the intermediate size.

The 64-celled sperm sphere, in the spermogonia stage, is interest-

ing, if we interpret it as a colony, for it reduces to a minimum

hitherto not met with, the possible size of a Volvoxr ageregation.

Klein regarded as such a minimum the small, sexually mature col-

onies formed within the parental coenobia in the special variation

of V. aureus before mentioned. These were about 100 uw in diameter

with approximately 180 to 200 individual cells, of which about a

third became spermogonia (“ Androgonidien ”’) and developed into

sperm platelets. The number 200 was the minimum, for the cells

of a free-swimming colony, which his earlier research gave him.

The diameter of these sperm spheres is not much less than 100 LL,

even for the smallest size. The three sizes range from about 80 pz

for the smallest to 180 » for the largest. But the cell number of

the smallest size is but a third of that in Klein’s smallest colony and

brings the cell number for a Volvox colony to but double that of

Eudorina. Indeed the resemblance of some of these smallest sperm

spheres, while in the spermogonia stage, to large compact colonies

of Eudorina which chance to be in some of my preparations is strik-

ing in the extreme. Even the size and shape of the cells and their

reaction to stains differ but little.

As to the genesis of these sperm spheres nothing is easier than

to trace it throughout all of its stages except the very first, and even

these are tolerably plain. As a glance at the plates will show, these

sperm spheres develop usually, though not always, in the same

coenobia and side by side with ordinary vegetative colonies.

(PI. x1, figs. 2 and 3; and pl. xu, figs. 7 and 8.) Their periods of

development, moreover, coincide as a rule closely, though not exactly.

For the examination of a very large number of cases discloses the

curious fact that well nigh the first indication as to the ultimate

character which the segmenting reproductive bodies within a coeno-

bium are to assume lies in the fact that certain ones are one or two

cell divisions ahead of the others, say, in the 16-cell or 32-cell stage

as opposed to two, four, or eight. These slightly more advanced

anlagen invariably, it seems, become the true vegetative colonies,

while the slightly laggard ones, seeming at first sight to differ in no

other respect from them, almost as invariably become the sperm

spheres. The instances confirmatory of this rule are very numer-

ous; those which deviate from it are few and usually, when they do

132 J. H. POWERS

occur, are great deviations—cases of irregular reproduction. As to

the enlarged cells which develop into these vegetative colonies, on

the one hand, and into the sperm spheres on the other, I at first

thought their appearance identical. And as we shall see later, in

no other form of Volvox is the parity (homology) between the three

forms of reproductive cells so close as in this type. Yet a further

study of these cells showed evidence that a slight inner differentia-

tion, accompanied usually with difference in size, had taken place.

These cells, in general, measure from 30 to 36 p. The more slowly

segmenting ones are the smaller by about one to three microns, their

nuclei are perhaps a trifle larger and, plainest of all, their cytoplasm

is denser, the transparent area between nucleus and chromatophores

being noticeably less. This somewhat denser character of the cells

is evident throughout all the developmental stages in the sperm

sphere.

These facts point strongly toward the interpretation that the

sperm spheres are not daughter colonies with an extreme and pre-

cocious sexual development, but, on the contrary, are really spermo-

gonia, checked before their final division into sperms, and stimulated

to undergo several generations of what we may call vegetative

division. The phenomenon is evidently comparable to the isolated

cases, to be described later, in which cells that had attained the full

size and character of ova then began vegetative segmentation ; al-

though what the result of such segmentation may be I cannot say.

Another fact indicates still more strongly this interpretation, v2z.,

the size of the cells throughout the entire development of the sperm

sphere. At no time does this fall below 6 », whereas in the young

vegetative colonies, after the first slower segmentation up to the

32-cell stage, it passes by rapid division to the small size of 3 p long

before the young colony has closed in to form a complete sphere.

From this time the cells of the young vegetative colony increase

slowly in size again throughout its entire growth period. (Pl. x,

fig. 10.) No such repeated division, rapid reduction, and slow in-

crease in size occurs in the cells of the young sperm spheres at any

stage, and it is the large size of the cells which renders them in-

stantly recognizable among the young daughter colonies, however

closely they may resemble these in general size and other respects.

(Pl. x1, figs. 2 and 3; pl. xu, figs. 7 and 8.)

On the whole, then, my general conclusion in regard to the rela-

tionship of this form, which perhaps 1 may well state now, is that

7 ’

Sn ee ee

ee eS ee a ee

NEW FORMS OF VOLVOX 133

it is not derived either from Volvox tertius or from the special form

of Volvox aureus, to both of which it shows a superficial resem-

blance, but from the typical Volvox aureus by a variation of an

opposite nature. Instead of precocious development with early

maturing sexual cells, there has been an increase in vegetative

growth, a retardation in the specialization, together with an increase

in the number of the sex cells, that is, of spermogonia and finally

of sperms. The facts which look the other way are the general

form of the sperm sphere, which makes it closely resemble a colony,

and cilia which it certainly bears at some stages of its development.

But the spherical form is characteristic in some degree even of the

sperm bundles; and cilia, likewise, probably occur, at least in some

stage, in all volvox cells. It will be seen that nearly all the other

characters of the form bear out, at least indirectly, this general view.

As to the number, size and structure of the sperms themselves,

they agree in the main with those of Volvox aureus. The single

sperm bundles contain invariably 32 sperms (pl. x1, fig. 12; pl. x1,

figs. 16 and 17), agreeing with Klein’s special form of V. aureus,

but disagreeing with both V. aureus and V. globator as usually

found. The numbers found in the former are usually 16 and 32,

the variation occurring in one and the same individual. Klein found,

rarely, as few as eight. V. globator commonly shows either 64 or

128 sperms in one platelet, while it is claimed the number may rise

to 256. As to the whole number of sperms simultaneously ripened

in a single coenobium, it greatly exceeds, in the form I am describ-

ing, anything hitherto found. The maximum number which I actu-

ally find matured in any one parent colony in my preparations is in

a medium-sizéd individual whose sole content consists of eight ripe

sperm spheres, all of the maximum size. Assuming each sphere to

be, as it looks, perfect, and hence to contain 256 sperm bundles, we

find the total number of simultaneously maturing sperms to be

8 X 32 X 25665536. I think this theoretical number very close

to the fact in this case. Judging, however, from the number and

size of young sperm spheres which I have seen in single coenobia, I

estimate that coenobia must occur bearing over twice this number.

In form and inner structure of the sperms themselves, they agree

fully with those of V. aureus. Overton’ has shown that the sperms

of V. globator and V. aureus differ markedly in structure: the for-

‘E, Overton: “ Beitrag zur Kenntniss der Gattung Volvox. Botanisches

Centralblatt XXIX: 30, etc. 1889.

134 J. H. POWERS

mer are slender with elongated nuclei and more or less laterally

attached cilia; the latter are less differentiated, being less elongate,

with spherical nuclei and terminal cilia. Careful attention to suit-

able bundles in figures 12, 16 and 17 will show that in this respect,

as in a majority of minor characters, this form relates itself closely

to V. aureus: the nuclei are spherical, the sperms compact and the

cilia terminal.

Turning to the more general features of the Volvox under con-

sideration, many other characters show themselves equally new, if

less striking. Thus the numbers and numerical relations of the

three sets of generative structures, eggs, sperm spheres, and vege-

tative daughter colonies, are again unique within the genus. The

maximum number of vegetative colonies is about twice that found

in any previously described form. V. globator shows regularly

eight parthenogenetic daughter colonies. V. aureus is highly varia-

ble. Four is often given as the common number, although this may

rise to six or seven under certain conditions. Overton, during long

search, found eleven the maximum. Oltmann’ gives twelve as the

maximum for the genus, although Klein had already, in his extended

research, raised the maximum in V. aureus to fourteen. Such num-

bers have, however, hitherto been wholly exceptional. With the

form I am describing they are frequent, and with the form I shall

describe at the close of this paper they may be multiplied five-fold.

In my examination of the fresh material, I remember distinctly hav-

ing counted 22 well-developed daughter colonies within one parent.

In my slides at present I cannot find so high numbers among the

developed daughter colonies. The greater proportion of the largest

coenobia may have been ruptured before my preparations were made,

But in the younger coenobia, both before and after birth, I find, not

infrequently, over 20 enlarged cells which must certainly in many

cases all become vegetative colonies. (Pl. x11, fig. 9 represents —

such a young coenobium with apparently 25, but really 24, repro-

ductive cells.) How high these numbers run may best be shown

by giving several concrete cases. A large coenobium, with ten

daughter colonies of maximum development before birth, the largest

measuring 210 by 240 p, gives the following counts: one daughter

colony showed but 16 enlarged cells, three showed 18, two 19, one

20, one 21, one 22, and one 23. Another coenobium, with nine

daughters of 205 to 225 m average diameter, showed four with 16

*Oltmann: Morphologie und Biologie der Algen. Jena; 1904.

NEW FORMS OF VOLVOX 135

enlarged cells, two with 17, two with 18, and one with 21. Another,

with six daughters of about 225 to 240 mw average diameter, gives

one with 17, two with 19 and three with 20. Yet another, with five

daughters of the maximum diameter of 250 p, gives the surprising

numbers of 15, 23, 23, 24, 24. The highest number that I find after

a moderate search is 25. There is no reason to think that in times

of exclusively parthenogenetic reproduction this number would be

reduced. More probably it would be augmented. In the case of

my material, however, the more richly reproductive coenobia seldom

remain exclusively vegetative.

In the number of ova or oosperms this Volvox again shows itself

independent. V. globator shows, according to Overton, 12 to 40,

usually about 30; according to Butschli, about 50. V. aureus pos-

sesses, according to Biitschli, never more than 8; according to Over-

ton, usually 5 or 8, sometimes 4 or even 3, and in partially vegetative

colonies, even one. Klein figures as many as nine. I find a number

intermediate between the maxima of V. globator and of V. aureus,

18 and 19 being the highest numbers at present found in my prep-

arations. Numbers approximating these are common, although the

number may fall in mixed coenobia as low as one. Obviously, here

again, this form is closest to V. awreus, in that the number of egg

cells has the same upper limit as has the number of vegetative col-

onies; while in V. globator the two show widely divergent relations.

In size, however, the eggs of this form are smaller than those of

V. aureus and resemble those of V. globator, being 51 to 54 p as

against 60 » for V. aureus. The walls of the mature zygotes are

again of an intermediate type. They agree with those of V. aureus

and differ from those of V. globator and V. carteri in that they are

plain instead of crenate or wavy. But they differ from those of

V. aureus and agree with those of V. globator in that the cell is not

asymmetrically but symmetrically placed within its inner wall.

(Pl. x1, figs. 5 and 6.)

The numbers of the sperm spheres also range very close to those _

of the ova and of the vegetative daughter colonies. Frequently a

single one is present in a parental colony bearing other reproductive

bodies as well (pl. x11, fig. 8), while the maximum number may rise

well up toward that of the ova or of ordinary vegetative colonies.

Fourteen sperm spheres, together with two ordinary daughter col-

onies in one maternal coenobium (pl. x11, fig. 7), seems to be the

highest number I have recorded, although I think higher ones occur.

136 J. H. POWERS

The close approximation to parity between the number of primi-

tive reproductive cells, which may develop into parthenogonidia, into

Ova, or into sperm sheres, is again a peculiarity of this Volvox. In

V. globator and V. cartert the anlagen of eggs and sperms are about

equal in number, but greatly surpass the maximum number of

parthenogenetic cells. In V’. aureus and apparently in V. tertius

it is the true ova and the parthenogonidia that run in parallel num-

bers, while the anlagen of the sperms may exceed these many-fold.

In the Volvox before me the numbers, at least the limiting numbers,

of all are closely parallel. This parallel in numbers, taken with the

close simultaneity in origin, which is the rule, and with the close,

though not entire identity in structure, seems to the writer to have

a not unimportant bearing on the vexed questions of homology be-

tween the different reproductive bodies in all members of the genus.

To make more complete as well as more detailed these relations

between the various reproductive bodies, I will give the results of

an examination of 100 independent coenobia taken serially as they

occurred on three of my slides, the only ones rejected being such

as were too young to admit certain determination of their reproduc-

tive contents. Of the 100 coenobia, 18 bore only vegetative daugh-

ters, the maximum number being 14 and occurring twice, the mini-

mum being 5 and occurring but once, the average being 9.5. The

number of exclusively female coenobia, 7. ¢., those containing ova

or oosperms only, was five, the numbers being 7, 8, 14, 15, 16; aver-

age 12. No coenobia occurred among the 100 showing exclusively

sperm spheres as reproductive contents, although such are not in-

frequent. (Pl. xu, fig. 11.) Likewise none occurred showing

sperms and ova without the presence of vegetative offspring as well;

such I have never seen. Over one-half of the colonies, 55, showed

the combination of sperm spheres with vegetative daughter colonies.

The maximum number of sperm spheres in one of these coenobia —

was II and occurred twice; the minimum number one, which oc-

curred fourteen times; the average was 4. Among the vegetative

daughter colonies of these same coenobia the maximum number was

14, occurring but once, and the minimum number I, occurring but

once; the average 7. The last possible combination of the repro-

ductive bodies was represented by 12 colonies which contained vege-

tative daughters together with both sperm spheres and eggs or

oosperms. ‘These were plainly larger than the average of the others.

The extremes among the eggs (or oosperms) being 1 and 7, the aver-

NEW FORMS OF VOLVOX 137

age being 3.3. The extremes among the sperm spheres being 1 and 8,

with the same average; while among the vegetative daughters

occurred the numbers 2 and 13, with an average of 7.2. These facts

are fairly typical, although the number of coenobia examined is of

course too low to represent other than an approximation to the real

numerical relations involved. The coenobia containing eggs alone

were never large; those containing all three classes of reproductive

bodies usually were large; those containing sperm spheres together

with ordinary vegetative colonies were of all sizes. A rather novel

fact, however, is the tendency of these latter coenobia to one extreme

or the other. One finds repeatedly a single sperm sphere among

a number of ordinary young (pl. x11, fig. 8), and not infrequently

the opposite relation (pl. x11, fig. 7), while evenly balanced coenobia

are less frequent. But two of the above 55 showed an equal num-

ber of ordinary daughter colonies and of sperm spheres. As to the

time of maturing of ova and sperms, the former are, in the rule,

fertilized and have become thin-walled oosperms before the sperm

spheres of the same parent coenobium have become fully mature.

This follows from the simple fact that the primitive sex cells start,

as a rule, almost simultaneously, and it is obviously a shorter route

to develop into a ripe ovum than to pass through both the growth

and the differentiating processes necessary for the production of a

complete sperm sphere with its multitude of differentiated sperms.

Notwithstanding this, instances occur of the simultaneous ripening

of ova and sperms, as in the coenobium represented in plate x1,

figure 3, where a-ripe ovum, apparently unfertilized, is in practical

contact with a colony of very fully developed sperm platelets. In

such cases as this the anlage of the sperm sphere undoubtedly started

ahead of that of the ovum. Such non-simultaneity, though as I

have said, not the rule, is not very infrequent among all the classes

of reproductive bodies.

As to the size of the adult colonies of this Volvox I can furnish

less exact data, all of the large colonies in my preparations having

been at least somewhat flattened. Measurements which I made on

the living material are not now at hand. They showed, however,

that the size exceeded the usual maxima given for any species of the

genus. Oltmann gives the highest maximum I have seen, I mm.

It is safe to place this as the upper limit of the present form. The

lower limit for colonies containing mature reproductive bodies is

much above the usual limit for V. aureus, 680 w being the lowest

138 J. H. POWERS

measurement I have found in my preparations, from which some-

thing may be subtracted for a little flattening, leaving as a minimum,

say, not less than 500 ». Klein found full-grown V. aureus as smail

as 175 yp.

In contradistinction to this large size of the adult coenobia, the

daughter coenobia are never as large or as fully developed as fre-

quently occurs in V. aureus. Doubtless this goes hand in hand with

the increased number of the daughter coenobia, for the same varia-

tions are carried to an astonishing degree in the next form I am to

describe. The maximum size of the daughter colonies before birth

is, as I have indicated above, about 250 pw, and the smallest free col-

onies are but now and then a little below this. The relative retarda-

tion of the daughter colonies, say at birth, is farther shown by the

fact that the parthenogonidia in this form never segment before the

daughter colonies containing them leave the parent coenobium.

This is very common in the ordinary forms. But in the type I am

describing segmentation of the reproductive cells only takes place

after considerable growth in the free state. Measurements on a

number of young colonies and their contained parthenogonidia show

this plainly. A few colonies are found above 400 p» in which but

part of the parthenogonidia are beginning to segment, but the ma-

jority show some segmentation at 350 ». Only daughter colonies

which contain very few reproductive cells, and which doubtless

escaped from smaller parents at a correspondingly smaller size, show

segmentation at 255 » or possibly at even less. The reproductive

cells—parthenogonidia, etc.,—within the largest unborn daughter

colonies measure but 15 to 18 p, while the regular size in the free

colonies that are beginning to show segmentation is, as I have said,

exactly twice this, viz., 30 to 36 ». These last figures are on the

whole very constant, but in rare cases a singular variation occurs—

a further retardation. I find in a very few coenobia cells in various —

stages of division (pl. x11, fig. 18), which, before dividing, have

reached nearly the size of ova and oosperms, 7. @., 51 ». Such phe-

nomena are very striking, and suggest the possibility that these may

not be parthenogenetic cells, but fertilized ova which, instead of

entering the usual resting stage, are developing at once. This is,

however, but conjecture, and is less probable than the hypothesis

of mere delay with continued vegetative growth.

Concerning the number of cells in the colonies, | am unable to

give accurate figures. To count the larger colonies correctly is not

NEW FORMS OF VOLVOX 139

possible, and estimates—e. g., according to the method of Klein—

all imply the sphericity of the colonies. As mine are flattened to a

certain though unknown extent, estimates become precarious. Such

as I have made, however, indicate that the number of cells is well

within the limits given by Klein for V. aureus. Indeed I think that

nothing comparable to his widest extremes—200 to 4400—occurs.

Certainly nothing is found approaching his lower limit for free col-

onies. Well grown daughter coenobia like plate x11, figure 13, show

about 1700 or 1800 cells, as may be verified by counts on the figure.

The largest colonies probably hardly exceed 3000. The extra large

size of these colonies is plainly due to the development of the gela-

tinous matrix.

Touching the finer structure of the colony, I shall omit such points

as are not characteristic of this form as well as those which my

preparations are not adapted to disclose. The size of the individual

cells in adult colonies ranges from 6 to Io p, most of them being

of the smaller size.1_ The distances separating them may be as great

as 50 p, but are usually from 28 to 40 p. The vegetative cells in

daughter colonies at birth are about 5 p» and increase to nearly or

quite full size by the time the parthenogonidia enter on division. In

shape these cells are more nearly spherical than is usually the case

even in V. aureus, resembling most of all the figure which Arthur

Meyer gives for V. tertius. Moreover, a relationship to this species

is further shown in the utter absence, as far as my investigation

goes, of connecting strands between the separate cells. Meyer failed

to find such connections only in V. tertius in the adult condition, the

younger colonies still showing them. I fail to find them at any stage

whatever. It is true I have not employed the technique resorted to

by Meyer, but I used reagents which ordinarily suffice, and on living

material. Many of my permanent preparations also seem well

adapted to the purpose, especially material fixed in the diluted Flem-

ming’s and strongly stained with rose bengal. This gives an admir-

* The larger cells are seldom wholly wanting in the larger coenobia, which

fact suggests the possibility that they are, in reality, another and much later

generation of reproductive cells. Such an interpretation would imply the

probability that this Volvox does not always die after the birth of one brood

of daughter colonies, but may live and develop another. My preparations

show a number of large coenobia destitute of ordinary contents, yet evidently

no longer ruptured. These, therefore, suggest the same possibility; while the

study of the second form of Volvox described is still more suggestive of such

a possibility.

140 J. H. POWERS

able fixation, seemingly devoid of shrinkage, and the stain takes

heavily in the cytoplasm while leaving, as most stains do, the matrix

perfectly colorless. Yet neither in old nor younger colonies at any

stage can I discover a trace of the connecting fibers, even under a

Zeiss apochromatic. The connecting strands in V. aureus are said

to be of about the same diameter as the cilia. Several observers

have failed to demonstrate them, but in some such cases their prep-

arations have failed to show cilia as well. In mine the cilia, both

within and without the matrix, are plainly visible with ordinary

magnification. Doubtless more investigation will be necessary

before concluding that these structures, supposably so characteristic

of the genus, may be entirely wanting. Meyer would not assume

their real absence in adult V’. tertius despite his elaborate technique

and his total failure to demonstrate them. Yet his refusal to accept

his own evidence seems to have been for rather theoretical reasons.

No one claims that the closely allied genus Eudorina possesses them.

Yet it shows the same phenomena of apparent coordination, etc.

Concerning the structure of the gelatinous walls and intercellular

substance I will say little, because I have not used the technique

which Meyer has shown to be necessary to discover the details of

the structure. Most of the reagents used by me were rather inef-

fectual in this regard. They did little more than emphasize the

structure which was faintly visible in the younger living coenobia,

vig., a separation of the surface into clearer, spherical areas, about

the individual cells, and grayer, intervening paths of connecting sub-

stance. This is shown to some extent in several of the figures.

(Especially pl. x11, fig. 10.) Such surface fields are wholly unlike

V. globator, but resemble those of V. aureus, and are identical with

those given by Meyer for V. tertius. Other indirect reasons lead

me to suspect that so far as the wall is concerned, V. tertius is the

most nearly allied to the form I am describing.

A few further words might now naturally follow, as to the sys-

tematic position of this Volvox, whether it constitute a new variety

(mutation?), species, or genus? But I will postpone this consid-

eration until I have briefly described the second form which came

under my observation.

I owe the discovery of this second form of Volvox again to my

pupil and assistant, Mr. George R. LaRue of Crete, Nebraska. It

was not in sexual reproduction at the time (September) when it was

discovered. But despite this fact it is little less noteworthy than

NS Se ee

NEW FORMS OF VOLVOX I4!I

the form just described and is especially interesting taken in con-

nection with it, as certain lines of variation which the first form

shows but moderately are in this one carried to surprising extremes.

Its size attracted the attention instantly, even when in the pond. I

judged that many of the colonies could not be less than 2.5 mm. in

diameter.1. In well preserved formalin material, showing a little

shrinkage, I readily find coenobia over 2 mm. in diameter and the

majority closely approach this limit. I examined thirteen colonies

taken at random before I found one as small as 1.8 mm. The aver-

age size of this Volvox, then, judged by its adult colonies, is more

than twice the size previously recorded for any species of the genus,

and three to four times the average size recorded for V. aureus.

The size, however, was dependent in this case even less than in the

preceding upon an increased number of cells. On the contrary I

find the number very low, often below a thousand even in the large

colonies and seldom much above it. They are about 12 » in diam-

eter, as against 7.5 to 8.5 » for the preceding form, and are separated

by varying intervals of from 50 » to as much as 200 p, in comparison

with 28 to 50 p» for the preceding form. If connecting strands of

cytoplasm of quite invisible fineness (I can make out no trace of

them) connect these cells, they are surely something of a phe-

nomenon.

A consequence no doubt of the large size and scattering cells of

these colonies was their extreme fragility. I lost most of the mate-

rial of my collection by entrusting it to a fixing fluid which has

often given me admirable results with related forms—leaving the

matrix of Eudorina, for example, wholly unharmed. But these

over-expanded colonies were quite unable to withstand its action.

Correlated also no doubt with the size of these colonies, though not

wholly thus explicable, was the most. surprising feature they dis-

closed, viz., the number of parthenogonidia or of vegetative colonies

they contained. In not a few cases both the parthenogonidia and

the young colonies had been partially or wholly destroyed by some

agency, possibly a parasite. In such colonies the remaining number

of daughter colonies, for example, might sink to any number, and

1 Professor Charles E. Bessey, of the botanical department of the Univer-

sity of Nebraska, informs me that he has also observed Volvox colonies in

this vicinity which were at least close to 2 mm. in diameter. This is interest-

ing as showing that the form I am about to describe is not an isolated or acci-

dental variation.

142 J. H. POWERS

in one instance one alone remained in a developed healthy condition.

Omitting such colonies, the count of ten instances gave the follow-

ing numbers of parthenogonidia or daughter colonies: 10 (very old

and probably not perfect), 15, 29, 30, 33 (not full number), 35, 41,

49, 49, 71. It is not probable that these numbers represent the

maximum,’ since the material examined was insignificant in amount.

But as they stand they are wholly without precedent, trebling, as

they do, the number found in the preceding form and multiplying

five or six-fold that hitherto found in any species or variety. As

indicated, ‘this high number of parthenogenetic reproductive bodies

may well be correlated with the large size of the colonies, or perhaps

with the wide sundering of the cells and absence or functional weak-

ness of the connecting strands allowing of less specialized concen-

tration of nutritive excess upon a few individuals. That it was not

purely a matter of size, however, is shown by the fact that the

reproductive bodies are not, as usual, confined to one-half of the

volvox sphere, but, in case of the higher numbers at least, were

distributed over two-thirds or four-fifths of the surface (pl. x1v,

figs. 19, 20, 21 and 23), simulating the distribution of the sperm

bundles in VY. aureus when these are very abundant.

With the great number of the daughter colonies and the feeble

nutrition of the flimsy parent coenobium was no doubt correlated

two of the remaining peculiarities of the form, 7. e., the insignificant

size of the daughter colonies and the complete retardation, until

after birth, of their reproductive cells. None of the daughter col-

onies which [ examined were above 150 p» in diameter. The major-

ity measured but 125 » or even less. I deemed these at first to be

merely very immature parthenogenetic progeny. But despite the

large size of the coenobia and the fact that some of them appeared

to have discharged a part or all of their contents, I could find none

with larger daughters. This led me to question whether this were

not the size at which these colonies were freed from the parent

sphere, and such seemed finally the inevitable conclusion. Unfor-

tunately but few immature, free colonies could be found. The small-

est, however, measured but 279 pw and yet was plainly an independent,

free colony. Its individual cells were 7 mw in diameter and were

already separated by intervals of 9 to 15 pw. (PI. xiv, fig. 25.)

*In the course of photographing the coenobia shown on plate xiv an-

other was found in the same small lot of material showing 78 parthenogonidia ~

or young colonies. (Pl. xiv, fig. 20.)

NEW FORMS OF VOLVOX 143

Not only, then, do the daughter coenobia here escape at a very

small size as compared with that of the adult, but they are equally

or even more retarded with regard to their reproductive cells. Not

one of the developing daughter colonies showed a trace of differen-

tiation of parthenogenetic cells of the next generation (pl. xtv, fig.

24), while in the smallest free colony, above mentioned (fig. 25),

they were just making their appearance, 30 of the component cells

having enlarged to from 9 to 10.5 pw, as compared with 7 mw for the

remainder. As we have seen, in ordinary species the partheno-

genetic ova differentiate, reach maturity, and even begin to divide

before the birth of the daughter coenobia containing them. In the

special form described above they reach. but half their full diameter

by the time the young coenobia are freed. While here in this form

they are not even distinguishable at this period. Their time of

maturity and division is indeed not until apparent maturity of the

coenobium, to judge by the material examined by me. A number

of almost the largest colonies (fig. 19) showed undivided, though

mature, anlagen; in others they were in early stages of segmentation

(fig. 20), while in the oldest coenobia (figs. 21 and 22), containing

young colonies of full size, there might still be present few or many

parthenogenetic cells of an altogether later generation. This varia-

tion constitutes almost as striking a change of habit for the genus

as did the mode of developing male gametes in the preceding form.

It would seem, too, of doubtful advantage, for these large fragile

colonies might easily be destroyed before the maturity of their late-

developing progeny. It is possible, as I have indicated before, that

more than one generation of young is here produced.

But one further peculiarity was noticed, wiz., the early age at

which the increase in gelatinous matrix began to separate the com-

ponent cells of the young daughter colonies. Ordinarily this takes

place hardly at all before their birth. Here it begins apparently as

soon as the young colonies are closed spheres. Unclosed colonies

showed the cells compactly pressed as usual, and this stage was

sometimes 60 » in diameter. Complete daughter colonies were

found, however, in the same parental colony, as small as 75 yp, in

which the separation had already begun, the cells being completely

rounded and none of them in contact. As above noted, in the small-

est free colony, of 279 » (pl. xiv, fig. 25), the intervals between the

cells have already increased to more than their own diameters. The

final result of this early-begun production of intercellular material

144 J. H. POWERS

is the production of colonies which, under the microscope, are

scarcely recognizable as volvox aggregations. Indeed many of the

older coenobia would not have been so recognized without study

(fig. 22). They had become practically motionless, were befouled

by adherent particles, their component cells, when not obscured,

showing only as pale green dots scattered about the surface.t| So

large, thin-walled and delicate were they that when resting in water

on the bottom of a watch glass they were flattened somewhat by

their own weight, filling more than the low power field of the micro-

scope and responding to every jar by vibrations and minor changes

of form. Yet even such colonies as these might contain at least

some healthy daughter aggregations. (Pl. xiv, fig. 22, near left and

lower margins. )

What of the affinities of these two forms of Volvox? It is em-

barrassing at the start that there are two of them. Are the two

distinct or only diverging forms of one species. They are certainly

different enough to be distinct. Yet there are many points of

resemblance, and the extreme variation of the second from the first

does but carry out some of the same lines of divergence which sepa-

rate this from V. aureus. It is safest to assume that the two forms

are closely allied. An added reason for this is found in the condi-

tions under which they occurred, the second form having developed

in much smaller numbers and in deeper water than the first. And

Klein has shown that V. aureus differs greatly in size, even in sim-

ilar and adjoining ponds, according to its sparsity or abundance.

The least crowding reduces its dimensions very much. This fact

explains something, if but little.

Taking, then, the two forms as, provisionally, one, what is their

relation to the genus Volvox and its previously described species?

Their relation to them is obviously close. Yet it is equally evident

that they do not fit into any previous specific definitions, and, no less,

that they quite transcend the definitions previously given for the

genus. This is true whichever of the two interpretations we put

upon the peculiar reproductive phenomena described for the first

form. Still this fact does not demonstrate this new-found Volvox

1 Without careful attention many of the component cells of the colonies

may well be missed in all the coenobia, except the last two, of plate XIV.

They may be seen however as small dots about 2 mm. apart. Others have

been retouched and somewhat distorted. Figures 21 and 22 show them only

near the center of each.

NEW FORMS OF VOLVOXx 145

to be even a new species. For generic distinctions may, no doubt,

be overstepped as easily by variation as may specific ones. Witness

the case, already mentioned, in the same family, that the new genus

(as supposed) Pleodorina intergrades perfectly with its well known

relative Eudorina. So these peculiar types of Volvox may well

prove to be only extreme variations (“ mutations,” if you will?

of Volvox aureus. Very pronounced variations they are, sharp and

clean and influencing numerous characters, well up to the mutation

standard. Should such prove to be the case, it would certainly be

the happiest consummation, for it would give us in this interesting

family of organisms a species of almost unheard-of variability and

thus open the way for promising experimental work.

In conclusion, I wish especially to thank Dean Charles E. Bessey,

of the botanical department of the University of Nebraska, for his

most courteous assistance in searching for the literature of the sub-

ject. The bulk of this lies on the botanical rather than the zoolog-

ical side.

146 J. H. POWERS

EXPLANATION OF PLATES

All of the figures are from photographs made by the simple, if some-

what unsatisfactory, expedient of a “ Brownie” camera inverted over a Zeiss

or a Leitz microscope. The unsightly quadrangular areas are the result, not

alone of an effort to save space in the plates, but of the fact that the camera

would not cover the full width of the larger microscopic fields. This latter

is indicated by the curved corners.

Plate XI

FIRST FORM OF VOLVOX DESCRIBED

Fig. 1. A medium-sized coenobium containing five egg cells (one parti-

ally obscure) and thirteen daughter coenobia, within which can be seen the

numerous anlagen of the next generation. Groups of minor organisms—

Euglena, Pandorina, Eudorina, etc.,—are adherent to the colony, or in the

field. Magnification about 56 diameters.

Fig. 2. A medium-sized coenobium containing ten immature oosperms,

one daughter colony (near lower margin) and three sperm spheres in dif-

ferent stages of development. The one farthest to the right is a medium-sized

sperm sphere showing platelets of mature sperms; to the left of this is a

large-sized sphere of dividing spermogonia, while above and to the left is an-

other medium-sized sphere of undivided spermogonia. Groups of minor or-

ganisms in the field and adherent to the colony, as in the last figure. In

upper right-hand corner portion of a young free colony with reproductive cells

still undivided. Magnified about 37 diameters.

Fig. 3. Small to medium-sized coenobium containing six daughter coe-

nobia, five apparently unfertilized ova, and two sperm spheres of fully ma-

tured sperms. Magnified 37 diameters.

Fig. 4. Small coenobium with 18 young oosperms. Portion of larger coe-

nobium with mixed contents and adherent groups of organisms above and

to right. Magnified about 37 diameters.

Fig. 5. Medium-sized coenobium (much obscured by groups of Trache-

lomonas, etc., as above) containing 17 mature oosperms. To left and below,

portion of still larger coenobium with very large daughter. The spherical

body in this latter is not related to the volvox. Magnified 65 diameters.

Fig. 6. Detail from the last. Magnified 319 diameters.

PLATE: Xt

. i cicx)S

am, * Sant it

PLATE XII

NEW FORMS OF VOLVOX 147

Plate XII

Fig. 7. Immature coenobium containing 14 young sperm spheres and two

ordinary daughters. These may be distinguished by their even outline, and

one by its ellipsoid form and larger size. Magnified about 74 diameters.

Fig. 8. Coenobium containing 17 ordinary daughters and one young sperm

sphere. Euglenas, etc., adhere to the exterior of the colony. Magnified about

50 diameters.

Fig. 9. Large daughter colony, with small portions of two others; seen

through the wall of the parent. It contains 24 undifferentiated reproductive

cells; there appear to be 25 but one is a similarly sized trachelomonas. The

four large cells which obscure a portion of the colony are extraneous. Mag-

nified about 108 diameters.

Fig. 10. Portion of young free coenobium which contained nine very

young daughter colonies and five very young sperm spheres. Four of the

former are shown, mainly in upper half of the figure; their cells are already

very small. Four of the young sperm spheres show in lower half of the fig-

ure, together with a part of a fifth. They show cells about the size of those

of the parent colony. The figure also shows the characteristic areas in the

gelatinous wall of the parent coenobium. Magnified about 217 diameters.

Fig. 11. Medium-sized coenobium containing eight of the largest-sized

sperm spheres, all with fully mature sperms. Extraneous groups of organ-

isms as before adherent to surface of coenobium. Portion of another coeno-

bium above. Magnified about 37 diameters.

Fig. 12. Detail from the last, 7. e., single sphere of sperm platelets situ-

ated, in last figure, near left side of the parent coenobium. Magnified about

217 diameters.

148 J. He POWERS

Plate XIII

Fig. 13. Large daughter coenobium, photographed through the wall of

the parent, the cells of which may be seen in the field as well as overlying

the younger colony. The daughter colony is seen from one end and appears,

therefore, spherical; its enlarged reproductive cells are somewhat greater

than those of the parent coenobium. Magnified about 319 diameters.

Fig. 14. Large-sized sperm sphere in spermogonia stage; from same

parent coenobium as the last. Magnified about 319 diameters.

Fig. 15. Medium-sized sperm sphere, the units of which are undergoing

their final segmentations. The cells within the individual groups vary in

size owing to a varying rate in segmentation which is, however, soon over-

come. A portion of a daughter colony shown also; both photographed

through the parent coenobium, some of the cells of which show in the figure.

Magnified about 319 diameters.

Fig. 16. Medium-sized sperm sphere of completely developed sperm plate-

lets. The wisps of cilia faintly visible in places, as also, in some cases, the

spherical nuclei of the individual sperms. Each bundle contains 32 sperms.

The whole aggregation was freed from the parent coenobium by pressure.

Magnified about 319 diameters.

Fig. 17. Another sperm sphere, like the last, but showing the beginnings

of disruption while within the parent colony, the cells of which are but faintly

in focus. Three sperm bundles had escaped from the aggregation and were

free within the coenobium; they are not shown. Nuclei plainly visible in

some bundles. Magnified 319 diameters.

Fig. 18. Detail from a large, but young coenobium showing unusual in-

stance of very large reproductive cells undergoing segmentation. Compare f

their size with the similarly magnified zygotes in plate xt, figure 6. Magni-

fied about 319 diameters.

PLATE XIII

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PLATE OALYV

NEW FORMS OF VOLVOX 149

Plate XIV

SECOND FORM OF VOLVOX

Fig. 19. Typical colony. Some of its individual cells are only seen upon

careful attention, showing a little less than 2 mm. apart. Others have been

retouched by the engravers so as to exaggerate their size and somewhat dis-

tort their form. The larger black dots, 30 or more in number, are the par-

thenogonidia, unsegmented or in early division stages. Only a portion of

them were in focus. Magnified about 17 diameters.

Fig. 20. A colony much like the last, but showing maximum number of

reproductive bodies found—78. A perfect count is not possible in the figure

but was easy under different foci, The two larger bodies are well grown

daughter colonies (fig. 24) ; of the others, one was an undivided parthenogo-

nidium, the others from the two-cell to approximately the 64-cell stages.

Many of the cells of the colony are nearly or quite invisible; others have

been retouched as in last figure. Magnified about 11 diameters.

Fig. 21. Large coenobium containing 49 well developed daughters, one

much smaller, unclosed daughter colony, and six young parthenogonidia

(easily recognizable by their strong staining reaction) of two to three times

the diameter of the vegetative cells. The last two classes of bodies do not

show in the figure. The daughter colonies are somewhat distorted by the

retouching done by the engravers. The wrinkling of this coenobium in the

preserving fluid displays the extreme tenuity of the colony membrane. The

individual cells are visible but here and there as minute dots. Magnified

about 17 diameters.

Fig. 22. An old coenobium characteristic of much of the material col-

lected. Most of the cells obscured by the light layer of adherent debris.

They may be seen, however, near the center of the colony where a few have

been slightly retouched. As sole reproductive content the colony contains

two large healthy daughters (below and to the left), four healthy, undivided

parthenogonidia and a few that were degenerating. Magnified about 22

diameters.

Fig. 23. Another colony much like figure 21, but containing 71 well de-

veloped daughters most of which may be counted in the figure. Only a few

of the individual cells of the colony may be seen. Magnified about 13

diameters.

Fig. 24. Daughter colony magnified to show early separation of cells and

entire absence of reproductive differentiation. Photographed through the

parent coenobium. A single cell of the overlying parent colony shows above

and to the left of the center. Magnified about 319 diameters.

Fig. 25. Smallest free coenobium found. Showed the beginning of the

differentiation of reproductive cells, 30 in all. Magnified about 108 diameters.

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PLATE XV

NEW APPARATUS, REVIEWS, ETC.

A PHOTOMICROGRAPHIC OUTFIT

By JAMES H. STEBBINS, Jr.

WITH PLATE XV

After experimenting for a long while on kerosene, Welsbach

burners, and acetylene as a source of illumination, I have finally

settled down to the arc light as the best all-round light for photo-

micrographic work.

My outfit consists of the following parts: a Fox automatic arc

light; a Zeiss optical bench supplied with a set of three Kohler

condensing lenses, specially designed for the use of the arc light,

and also manufactured by Zeiss; a water cell for cooling the

rays; an iris diaphragm for regulating the size of the cone of light;

a unicum shutter; and two cells for holding colored solutions,—in

other words, two ray filters. The microscope used with this outfit

is a Zeiss photomicrographic microscope, with a large short tube,

designed expressly for this purpose, and permitting the use of the

Zeiss planars for very low power work. The coarse adjustment of

the microscope is connected by means of a belt passing over one of

milled heads of the coarse adjustment to a pulley connected with

a long brass rod on the underside of the microscope and camera

stand and terminating at its farther end with a large milled head,

- by which means the microscope may be coarsely focused from the

farther end of the camera. The fine adjustment is operated in a

similar manner from the farther end of the camera. The camera

in use in my outfit is one that was especially made for me by Folmer

and Schwing. It is an 8x 1o inch camera, with a bellows drawing

out to about five feet, and is so arranged that the bellows can be

extended from either end.

The camera-bed is extended out far enough in front of the cam-

era to hold the platform upon which the microscope rests, and I have

found that by this means vibration is reduced to a minimum,

The camera, microscope and optical bench all rest on a single,

light, long, wooden table, and although I have worked in localities

*Cf. Minutes of the meeting, p. 159, and announcement, p. 159.

152 NEW APPARATUS, REVIEWS, ETC.

‘

where there was much vibration from passing trucks, trolleys, etc.,

yet owing to the peculiar arrangement of my outfit, I have had but

very little trouble in this particular direction, even with very high-

power work.

It will hardly be necessary for me to say anything in regard to

the plates and developer I use, as each person has his pet plates and

formulas, but in the work which I have to do, I have found that

the Cramer isochromatic slow and the Cramer trichromatic plates,

using glycin as a developer, have given the best results.

I trust that this rather erratic description of my photomicrographic

outfit may be comprehensible.

THE PIETZSCH MICROTOME

By Epwarp P. Do.LBy

In undertaking to improve the rotary microtome was realized the

necessity of making it as simple in construction as efficient, and as

practical as possible, and we feel satisfied that there have been added

many appreciable advantages to the excellent invention of Professor

Minot.

First of all it was endeavored to construct a knife holder which

would serve to operate both paraffin and colloidin preparations, thus

doing away with the expense and inconvenience of an extra holder,

while moreover affording the advantage to the operator of being

able to read the angles of inclination, so that the most favorable

angles for different kinds of work may be noted, and taken when

doing again the same work.

Another important advantage of this microtome is that the active

part of the knife may be restricted in order to obtain the greatest

rigidity and to use the entire length of the edge before resharpening.

It will be well to notice that the knife is clamped against a three-

point plane, logically the only secure clamping device for a honed

knife, which only by accident is a perfectly straight plane. Even

a new knife is seldom of a perfect, straight plane, an old one never.

The object carrier is exceedingly rigid and simple, though it per-

mits the adjustment of the object in every plane as well as the most

complicated heretofore constructed, and has besides the advantage

of being clamped by only one screw instead of two or three, and

NEW APPARATUS, REVIEWS, ETC. 153

also that of allowing a horizontal adjustment of the preparation in

order to bring it close to one of the knife clamps.

The coarse adjustment is certainly one of the most convenient

devices which has been lately invented for microtomes. It is a great

time saver, effecting a rapid advance or retrograde movement of the

object carriage in order to bring the preparation quickly, though

exactly, near the knife when beginning, or to cut off worthless sec-

tions, etc., without releasing the automatic feed.

Moreover all gearing is reduced, both as to size and to number

of parts, as much as possible and either so encased or so placed as

to guard against danger to the operator and important mechanisms.

The automatic feed is entirely new, and the most accurate ever

constructed, in which when working the shock on the feeding screw

and its bearing is entirely avoided. It is provided with a worm,

thus working with the greatest smoothness and accuracy up to the

present time attained. It is an inclined plane gliding on another

inclined plane,—the worm gear,—and therefore there is nearly no

friction or wear—wear being proportioned to friction. It is the

only device which by the use becomes more accurate, as the two

inclined planes gliding always on each other become more and more

smoothed by the wear. This feed is the only one which advances

only when the object is clear off the knife. Finally, only the worm

gives the advantage gained by cutting by hand of producing the

finest sections of such different thicknesses as may be required.

NEW MODEL PROJECTION APPARATUS—BAUSCH

AND LOMB LANTERN “D”

The value of the projection lantern for educational purposes is

becoming daily more apparent and has resulted in a demand for a

simple and inexpensive lantern for use in schools of all grades, for

colleges, for lecture platforms, and for societies. This demand has

been increasing rapidly and at last has been met.

The Bausch and Lomb Optical Company have just put upon the

market a new model, which while it is simple and inexpensive, is

at the same time efficient, portable, of scientific accuracy, and of

pleasing design.

The most important characteristic of this lantern is its converti-

bility. The various parts are made interchangeable so that one lan-

154 NEW APPARATUS, REVIEWS, ETC.

tern may be used for ordinary, micro, vertical, or opaque projection

with comparatively little manipulation. The lathe bed construction

has been used in this model, thereby securing rigidity, stability, and

perfect alignment so that no recentering is required. The lantern

is provided with a cooling cell which makes it possible to use living

objects. Without this device they would be unable to stand the

sntense heat. The lamp box is of unique design, which renders it

very efficient in carrying off the heat generated. The carrying case

may be used as a base or as a shipping case. An entirely new con-

struction has been used in the dissolver, which gives a perfectly

dissolving view. The change from ordinary to micro-projection 1s

easily effected by simply removing the front board carrying the pro-

jection lens and replacing it by the one carrying the microscope, the

front boards being held securely by the spring catch. The micro-

scope is of new design, the coarse adjustment having an unusually

long range and the sliding arrangement of the body tube furnishing

an additional means of adjustment for very low power lenses.

Where the high price has hitherto prevented a very extensive use

of the projection lantern that objection has now been removed with-

out in any way effecting the efficiency of the apparatus.

A new supplement to the Bausch and Lomb Projection Catalogue

has been issued describing this new model, and this will be sent on

request to any one desiring further information with regard to this

lantern.

THE MICROSCOPY OF VEGETABLE FOODS

Noteworthy among recent books of interest to all microscopists

ss one entitled The Microscopy of Vegetable Foods, with special

reference to the Detection of Adulteration and the Diagnosis of

Mixtures, written by Dr. Andrew L. Winton, in charge of the

Analytical Laboratory of the Connecticut Agricultural Experiment

Station, with the collaboration of the eminent Austrian authority,

Dr. Josef Moeller, and published by John Wiley and Sons, of New

York City.

The dearth of literature, in English, on a subject of such scien-

tific and technical importance is noteworthy, and the present work

most timely. It is designed for the use of the food analyst, the

agricultural chemist, the pharmacist, and others engaged in the

NEW APPARATUS, REVIEWS, ETC. 155

examination of foods, as well as the physician who may be called

upon to identify vegetable substances in stomach contents and feces,

and the author is to be congratulated upon the very happy manner

in which he has presented his material.

The work is divided into the following parts: I. Preliminary

Equipment, Methods and General Principles. II. Grain: its Prod-

ucts and Impurities. III. Oil Seeds and Oil Cakes. IV. Legumes.

V. Nuts. VI. Fruit and Fruit Products. VII. Vegetables.

VIII. Alkaloidal Products and their Substitutes. IX. Spices and

Condiments. X. Commercial Starches. General Bibliography.

Glossary. The descriptions of edible fungi in Part VII, of leaves

in Part VIII, and of barks, roots, leaves, and flowers in Part IX,

and the analytical key to the commercial starches in Part X, are by

Professor Moeller.

The text is arranged in a very systematic manner, the treatment

of each substance including a brief statement of its source, the dis-

tribution of the plant, its varieties, and the botanical characters of

the part used, followed by a detailed account of the histology of the

part and its diagnosis, ending in every case with a bibliography.

The work is profusely illustrated and numerous analytical keys add

to the value of the book as a reference guide. While the evident

intent has been to make the text in the highest degree practical, this

has been secured without in any way interfering with a thoroughly

scientific treatment of the subject.

WATSON & SONS’ INSTRUMENTS

A feature which is worthy of special notice in connection with

recent introductions by the opticians, W. Watson & Sons, of 313

High Holborn, London, England, is a long range of movements on

mechanical stages.

With the rapidly increasing demand for instruments affording

facilities for systematic examination of blood streaks, bacteriological

specimens, etc., a necessity has arisen for mechanical stages giving

a very long traverse. In Watson & Sons’ instruments this is accom-

plished in a strikingly efficient manner. In one stage which they

call the ‘‘ Scop” the milled heads controlling the vertical and hori-

zontal movements are mounted on the same stem, on the Tyrrell

system, and set at an angle on the right-hand side of the stage com-

156 | NEW APPARATUS, REVIEWS, ETC.

pletely out of the way and yet in the most convenient position for

working. With this stage any desired length of traverse can be

provided, but the ordinary stock pattern gives two and one-fourth

inches. |

The other stage is the one fitted to their “ Bactil”’ microscope and

might be described as a semi-detachable stage. The vertical move-

ment is operated by a rack and pinion and the upper travelling plate

is exactly like that of the plain stage of an ordinary microscope.

The horizontal fitting is carried completely as a separate mechanism

and is attached to the plate just referred to by means of two thumb-

screws. When in position the object can be carried horizontally

rather more than two inches. This stage is peculiarly convenient

because it combines all the advantages of the best type of mechan-

ical stage, and when the horizontal mechanism is removed the clear

surface of an ordinary plain stage instrument is at once available

with the added convenience of a vertical mechanical movement.

These are only two items from a catalogue, which comprises 178

pages, that is published by W. Watson & Sons, and which we are

sure it would interest the majority of our readers to apply for and

read. Messrs. Watson offer to send this catalogue free by mail to

members of the American Microscopical Society who may apply

for it.

ROSS MICROSCOPES

Ross microscopes have always been distinguished for possessing

the essential principles of first-class instruments. Superior work-

manship and strict attention to detail in construction, the results of

long experience,—the firm having been established as far back as

1830,—have given them their universal reputation. Careful study

of modern requirements has led to the production of the up-to-date »

Ross “Standard” model. It embraces the most recent ideas and

many improvements not to be found in instruments of other manu-

facture. The “ Standard,” with two achromatic objectives, may be

purchased at a price which is very moderate for a microscope of this

type, while other objectives, focusing swing-out substage, con-

denser, mechanical stage, and other accessories, are readily added at

any time, with equal efficiency, so that the same instrument can be

subsequently used for the most advanced work.

Messrs. Ross, manufacturing opticians, of 111 New Bond Street,

NEW APPARATUS, REVIEWS, ETC. 557

London, England, will forward free by post, on application, a very

concise little explanatory booklet.

-—

ANNOUNCEMENT BY THE SECRETARY

From a number of our members have come to the Secretary ex-

pressions showing a desire for a few pages in our Transactions of

a less formal character where may be inserted brief notes on micro-

scopical technique, modifications of microscopical apparatus, or on

investigations not of so serious or extended nature that they seem

to justify the preparation of a formal contribution. Especially has

this desire been evident recently, since this volume has become the

only publication in this country, of any kind, devoted to microscopy.

Moreover, it has seemed to us that much of good would accrue

both to our members and to our advertisers if in such pages the

latter could be offered the opportunity to call attention to new pieces

of apparatus or new modifications of old, or to present other matter

which would enable members, especially those more isolated, to keep

in touch with the progress in microscopical appliances and methods.

Accordingly at the Sandusky meeting the Secretary was instructed

to set aside a limited number of pages for brief notes on methods,

on new apparatus, books, etc., to be contributed by members and

advertisers. To both the invitation is given to contribute freely,

but at the same time the request is made that the contributions be

brief and to the point, that they be given in the spirit of helpfulness.

To advertisers the suggestion is also made that to offer that which

is essentially advertising matter will defeat the very end in view,

that of bringing them into closest sympathy with the individual

members of the Society. The generous support accorded by those

who make use of our advertising pages is a very material aid in the

publication of our Transactions, the success of which is vital to the

welfare of the organization; every member should feel a personal

interest in our advertisers, and a personal responsibility in the mat-

ter. Each one is performing a service for this Society when he

bestows his patronage accordingly, and when in his dealings he

makes it known that he is a member of this Society.

‘hed ne § at

ee

PROCEEDINGS

OF

The American Microscopical Society

MINUTES OF THE ANNUAL MEETING

HELD AT

THE LAKE LABORATORY, SANDUSKY, OHIO, JULY 5 to 8, 1905

The twenty-eighth annual meeting of the Society was called to

order in the Auditorium at the Pavilion, Cedar Point, Sandusky,

Ohio, at 11.15 A.M., Wednesday, July 5, 1905, for a business session,

with President Henry B. Ward in the chair. The report of the

Secretary was presented informally. A suggestion included in the

report to the effect that a limited number of pages in the Transac-

tions be set aside for brief notes on methods, on new apparatus,

books, etc., to be contributed by members and advertisers, was favor-

ably received and the Secretary instructed accordingly. The Treas-

urer being prevented by illness from attending, no report from him

was presented, but on motion he was instructed to close the books

at the end of the fiscal year and to submit them to an Auditing Com-

mittee to be appointed by the President. The report of the Cus-

todian was deferred to a later session and the Society adjourned

at 11.45 A.M. |

SECOND SESSION

The second session was held at the Lake Laboratory of the Ohio

State University, being called to order by President Ward at 2 P.M.

of the same day.

The Society was first welcomed by Prof. George L. Dietrich on

behalf of the city of Sandusky. Then by Prof. Herbert Osborn

on behalf of the Lake Laboratory, the University of Ohio, and the

State Academy of Science. In his address Professor Osborn called

attention to the associations connected with this locality, which was

160 PROCEEDINGS OF THE

selected by a former prominent member of this society, Prof. David

S. Kellicott, as the site for the laboratory. These addresses were

warmly responded to by President Ward, who, after having fittingly

acknowledged the sentiments expressed, continued as follows:

I cannot refrain from saying a few words with regard to the pecu-

liar propriety of the meeting place in which we are assembled. It

is, of course, true that for many years this Society has devoted con-

siderable work, through its members and officers and through the

limnological commission which it has organized, to the furtherance

of work in our own country on fresh-water biology. This alone

would be sufficient cause for our meeting at this point, but there are

even more cogent reasons.

In the first place, we are met on the invitation of the first perma-

nent lake laboratory on the American continent. To be sure there

have been temporary institutions maintained at various points, one,

indeed, by the United States Government, for several years, on

Put-in-Bay Island, not far removed from this very point, but the

temporary organization of this laboratory considerably antedates

most of these other enterprises, and the building in which we are

met was planned and erected for the specific purpose of constituting

a permanent station for fresh-water biology. Its inauguration under

the auspices of the University of the great state of Ohio insures it

the permanency it deserves, and promises to science the inestimable

results that are sure to follow the careful and continued investiga-

tion of lacustrine biology.

But there is a personal reason far deeper and in my opinion more

powerful, a reason which of itself alone would not only justify but

demand our meeting in this place. This annual session may be

rightly said to be a recognition of the work done by Prof. David S.

Kellicott for our Society and for American biology. He was not

only one of the founders of the American Microscopical Society

but one of its first secretaries, and as an editor of a series of six

volumes he made our annual report more than a mere pamphlet

record of proceedings. Both by the papers which he solicited and

no less by those which he himself contributed, was he successful

in raising our annual volume to a reputable position among the pub-

lications of learned societies. As twice president of the American

Microscopical Society, as a constant attendant upon its meetings

and worker in its councils, the Society owes him a debt of gratitude

AMERICAN MICROSCOPICAL SOCIETY 161

which it can never fully measure, and which it is both our duty and

our privilege to recognize to-day. It was Professor Kellicott who

first conceived the idea of a laboratory on Sandusky Bay, and who

for many years without assistance conducted a little experiment sta-

tion here on the shores of Lake Erie. The results of these investi-

gations are to be found in his many contributions to our own and

other periodicals, but it was he also who conceived of this perma-

nent laboratory, who sought to win friends and support for the

realization of his idea. It was he who in a true sense laid the

unseen but stable foundations on which arise to-day a worthy organ-

ization for the prosecution of biological research, and it is not too

much to say that if he had planned less broadly or founded the walls

less firmly and deeply we should not be able to enjoy so fine a

building or so comprehensive and permanent an organization for

biological research as that in which and with which we are met

to-day.

The following papers were presented at this session:

Prof. Henry B. Ward: “ Trypanosomes and Disease.” Discussed

by Prof. Osborn, Prof. Walton, and Mr. Slocum.

Prof. F. L. Landacre: “ Chlamydomonas and its effect on the dis-

posal of Sewage.” Discussed by Dr. Slocum, Mr. Pflaum, Prof.

Henry B. Ward, and Dr. R. H. Ward.

Prof. L. B. Walton: “Some Notes on Microscopical Methods.”

Discussed by Mr. Pflaum, Prof. Henry B. Ward, Prof. W. F. Mercer,

Prof. Osborn, Prof. W. F. Kellicott, Dr. Slocum, Dr. R. H. Ward,

and Prof. Wolcott.

THIRD SESSION

The third session, in the evening, was occupied by the presidential

address, delivered in the Auditorium of the Carnegie Library, San-

dusky, by Prof. Henry B. Ward, on the subject: “ The Relations of

Animals to Disease.” It was a most interesting fact that Dr. R. H.

Ward, father of Prof. Ward, and first president of the Society, pre-

sided. The admirable address was listened to by a large and appre-

ciative audience, including many of the physicians of Sandusky and

_ vicinity.

FOURTH SESSION

The fourth session was called to order at the Lake Laboratory at

9.30 A.M., Thursday, July 6, and the following papers were presented:

Prof. R. H. Wolcott: “A New Mite Parasitic on the Carnation.”

162 PROCEEDINGS OF THE

Mr. B. H. Ransom: “ Haemogregarines (Haemogregarina py-

thonis) in a snake tick (Amblyomma sp:).” Discussed by Prof.

Henry B. Ward, Prof. Osborn, and Prof. Walton.

Mr. Chas. Brookover: “The Preservation of Protozoa.” Dis-

cussed by Prof. Henry B. Ward and Prof. Wolcott. |

Prof. B. L. Seawell : “ The Biological Survey of the Pyrtle Spring

Lakes, Missouri.” Discussed by Prof. Henry B. Ward, Mr. Pflaum,

Prof. Walton, Prof. Osborn, Prof. Bleile, and Prof. Kellicott.

Mr. B. H. Ransom: “ A Nematode (Probstmayria vivipara) in

Horses, hitherto unreported in the United States.”

Mr. B. H. Ransom: “ A larval cestode (Dithyridium cynocephalt)

‘1 the muscles of a marsupial wolf (Thylacinus cynocephalus).”

Prof. F. L. Landacre: “A Bibliography and Check-List of North

American Protozoa.” Discussed by Prof. Henry B. Ward, Mr.

Ransom, Prof. Osborn, Prof. Wolcott, and Mr. Pflaum.

Prof. Henry B. Ward: “The Erection of a Memorial to Ernst

Abbé.” In presenting this subject and urging upon the members

the appropriateness of contributing to the fund to provide this memo-

rial, Prof. Ward said:

It is peculiarly appropriate that the Society should be called upon

to contribute towards the Memorial Fund for Professor Abbé.

Through his inventions the work which we are enabled to do with

the microscope has been increased many fold in accuracy and it is

not too much to say that his work exceeds in value that of any

one man since the discovery of the instrument. Not only in inven-

tion, but also in the organization of the business and in the inception

of the great cooperative undertaking known as the Carl-Zeiss-

Stiftung, has Abbé been the genius of the work, and yet his name

appears nowhere.

After a discussion of the matter it was moved to authorize the

officers of the Society to act as a committee to collect funds for the

purpose proposed and the motion prevailed. The Society adjourned

at 11.45.

FIFTH SESSION

The members were again convened at the Lake Laboratory at

2.40 P.M.

The appointment of committees was announced as follows: Au-

diting committee on the Custodian’s report: Prof. W. F. Mercer,

Mr. E. P. Buffet ; committee on resolutions : Prof. C. H. Eigenmann,

Prof. B. L. Seawell, Mr. B. H. Ransom.

AMERICAN MICROSCOPICAL SOCIETY 163

The following were on motion elected members of the nominating

committee: Dr. R. H. Ward, Prof. A. M. Bleile, Mr. Magnus

Pflaum, Prof. C. H. Eigenmann, Prof. H. S. Osborn.

A symposium followed on animals as parasites, participated in by

Mr. B. H. Ransom, who showed a large and valuable collection of

specimens loaned by the Bureau of Animal Industry of the U. S.

Department of Agriculture, and Prof. Henry B. Ward, who exhib-

ited a beautiful collection of slides and specimens contributed by

numerous members and other persons from various parts of the

world. The discussion which followed was joined in by most of

the members present. The members were adjourned at 4.15 P.M.

to gather again later in the evening at a very delightful compli-

mentary luncheon tendered the Society by the Ohio State University.

SIXTH SESSION

The Society was called to order for the sixth session at the Lake

Laboratory at 9.30 A.m., July 7.

The report of the Custodian was presented and with it the report

of the committee appointed to investigate the condition of the

Spencer-Tolles Fund investment, which showed the money to have

been most wisely and carefully invested by the Custodian. The

report of this committee was accepted and approved and the report

of the auditing committee on the Custodian’s report was also made

and accepted.

The matter of sale of Proceedings and the increase of the Spencer-

Tolles Fund were both referred to the Executive Committee.

The report of the Spencer-Tolles Fund Committee was read, rec-

ommending the appropriation of Grant No. 4 to Prof. Henry B.

Ward to assist in the publication of an extended paper on fresh-

water investigations during the last five years. On motion the grant

was allowed.

The report of the committee on resolutions was read, congratu-

lating the Society upon the choice of meeting-place and expressing

' the thanks of the Society to the Ohio State University, to the Ohio

Academy of Science, and to the citizens of Sandusky for the many

courtesies extended; congratulating the State and the University

upon the establishment of the Lake Laboratory and its present suc-

cessful administration; and expressing the thanks of the Society to

the officers and investigators at the Laboratory for their generous

and abundant hospitality.

164 PROCEEDINGS OF THE

The report of the nominating committee was presented and the

following officers elected:

President, Prof. S. H. Gage, Ithaca, N. My

Vice-Presidents, Dr. A. M. Holmes, Denver, Colo/: Prof. Haas.

Weber, Columbus, O. |

Treasurer, Mr. J. C. Smith, New Orleans, La.

Members of the Executive Commitee, Prof. M. J. Elrod, Missoula,

Mont.; Prof. Herbert Osborn, Columbus, O.; Mr. B. H. Ransom,

Washington, D. C.

A vote of thanks was tendered President Henry B. Ward. There

still being time left in the session, the following papers were pre-

sented :

Prof. R. H. Wolcott: “A Second Report on the Biology of the

Sand-hill Lakes of Nebraska.”

C. H. Eigenmann: “ Factors Controlling the Distribution of

Fresh-water Forms, as illustrated by Collections from Patagonia.”

With the second paper were presented resolutions addressed to

the President of the United States calling his attention to the desira-

bility of a comprehensive biological survey of the Panama canal

zone and the immediately adjoining territory for the following rea-

sons: Panama is a point of strategic importance in the study of the

distribution of fresh-water organisms in South and Middle America. —

It is certain that the Pacific slope fresh-water fauna of South and

Middle America was derived from the Atlantic slope fauna. The

Isthmus of Panama is one of the possible routes of migration. The

Panama Canal, when completed, will destroy natural barriers and

cause the faunas of the two slopes to mingle to a great extent. It

will thus permanently obliterate the natural and primitive conditions

and it is highly desirable that a biological survey of this region be

made before the completion of the canal.

The following papers were read by title:

Mr. Clark Bell: “The Relation of the Microscope to Medical

Jurisprudence.”

Mr. J. C. Smith: “ On the Occurrence of an Amoeba sp. parasitic

in Volvox globator.”

Mr. J. C. Smith: “On a Probable Sexual Life History of the

Amoeba.”

Mr. J. C. Smith: “ Three Species of Synchaeta new to Science,

from the brackish waters of Lake Pontchartrain, La.”

AMERICAN MICROSCOPICAL SOCIETY 165

Dr. M. D. Ewell: “A Contribution to the Microscopic Study of

Pen and Ink Lines.”

Prof. C. E. Bessey: “ Structure and Classification of some lower

Algae.”

Mr. W. H. Seaman: “ Microscopical Reminiscences.”

Prof. Henry B. Ward and Dr. D. C. Hilton: “A new Bothrio-

cephalid Tapeworm of Man, from the United States.”

The Society then adjourned sine die at 12.30 P.M.

In the afternoon of this day the members enjoyed a complimen-

tary excursion tendered by the Lake Laboratory. The route was

to Sandusky and from thence to Johnson’s Island, and a most de-

lightful outing and collecting trip it proved to be.

In the evening a social session on the beach was participated in

by all of those present, who thoroughly enjoyed the good fellowship,

the stories, the jokes, and the songs, every member being obliged to

participate in some manner in the impromptu program.

On the following day, July 8, the meeting was brought to a

fitting conclusion by a second very enjoyable excursion, also ten-

dered the Society by the Lake Laboratory, to Kelley’s Island and

Put-in-Bay.

Rosert H. Wotcorrt,

Secretary.

166 PROCEEDINGS OF THE

TREASURER’S REPORT

FROM MARCH 30, 1905, TO JUNE 18, 1906

DR.

By Balance from last Report ......... see cesses tree ees $ 29 61

To Membership tes) (TODS 1 .iaut cota etislg Shue isin: sien eth Wipiagciede ae ee nO

To Membership dues, 1904 ......-. cc escecccescveieeress (22 00

To (Membership) Ques) \BOO5 (hte go ele tis eile b's wo ali oley sie mp Aco eer

To Membership (dies) TOO) isi validate ewes nbs wmeniae oe 91 OD) OO

To Membership dues, 1907 .........scececessceceseecces 4 00 386 00

To UA AIS SHORE WREES |e Gitctale Maca te Wiel ISM mb. be ik lecabere dep ole ote eee 51 00

Ts ‘Subscribetss | Vike | Vee Va iatene mimi st ku de Obs on elo im wat be robin 2 00

To Subscribers VOL (ure W Lids ennai viata nee» binsw viele ele ala letelaielw ld EO ae 32 00

TO LA CVEPHSeES) VOL UR es Vibiabrdibk isin a 0's vaibelewtesiaaia ely I2 00

TOW OlMMES BOL eee iieia Wie oitiela ie lw mcoih feb sie h Sle Colm foie 4 65

DT ADOT ATTONN Whose Gale pun ote Ceci ay Rascal etd beepers wlieln ty os) Sieh ellos 6 Sui I 00

$516 26

CR.

By Postage, Secretary .......ccces sees cece seer eeecens $216

By Postage, Treasurer .........c cece ee ce ec te cere er ences 15 00 $17 16

By Expressage, Secretary ........eceeeee cece cree ee ceees 35 42

By Expressage, Treasurer ........ce ce ceee cece cece teers 2 70 38 I2

By Stationery and Printing, Secretary .........---+-+e+s 12 20

By Stationery and Printing, Treasurer ........-++++++e-- 6 60 18 80

By Typewriting, Secretary ......... cee cece cece ec eeceees 36 30

By Sundries, Secretary .......c cee ee cece ee cece ener eens 3 00

By Plates for Vol. XXVI ....... see ee cece ence ecenecees 34 43

By Fare to Buffalo Meeting, Secretary ........-..++.0+- 25 00

By Fare to Sandusky Meeting, Secretary ...........++-- 24 20

By Balance on hand ..........eeeececece cece cecns 319 25

; $516 26

J. C. Smrrsz,

Treasurer.

We do hereby certify that we have examined the Treasurer's books and

the vouchers submitted therewith and find the accounts correct in accordance

with the foregoing statement.

Henry B. WArpD,

CHARLES FORDYCE,

Auditing Committee.

AMERICAN MICROSCOPICAL SOCIETY 167

CUSTODIAN’S REPORT FOR YEAR ENDING JULY

I, 1905

SPENCER-TOLLES FUND

meported at buffalo Meeting cig vedi ded ac eleven bes $1916 59

BAPPRICHUSHITOCEIVEU Cou che hmncrrety ante Ut ean n AD nl Aa feat nie 2 125 69

PCO LE TUCECCING Sy lor etl oink tees valde eile cay tues 60 00

PEPIDINTONE, a whee diablo s eure Oe Ue eee ode ale as Seed 5 00

1 CTA cer Ne hrc Bl oR, ah BLE G9 MUA Sn Lo IG SY $25 00

CSS ME MHER SES Y Altay oc dan Wma uA ie Livia was altic daa 5 00 30 00

PLGTALTINVEStEG Stas terme tate ie se glarstar wk 2077 28

MDA Ale OM UA cinta cles ciel ete tales iu Oi, he ylalakins 3 81

EE OUTRO LUUETC Mant iscctere ie Ail ts cole Ni SN aN Tithe t $2081 09

RaVrOSSMITICSCASE: KUTINs YEAR GUN Cu uci eiel eel bel'és sarclaldalitere 194 50

PECTIC REASG CUEING VEAL) oui ay Uisie ci eiethid aialele\clclwal oye Cass 164 50

ANNUAL GROWTH

Year Increase Total

CREE ry aie ga tale a} Bide MOA Cuig treme ee ewe ok $ 60 20

Coe a ES pli MEN AE AY AE RS Lag) 1 BA aE Ie aL PL ON RS av $ 25 00 85 20

MRRP tL oy 4.0 bie Stok ev Hiare e aividlakdiae aldaen eats IO 00 95 20

OS) ah LR CURR a A i GS Rg 52 66 147 86

CEC TES oe OCT NUS han ann AE ORS RO AN ta Cs A 76 00 223 86

SPRUE ROTO tis ar Le arial sila Ui Wats thy batt aeaie tea hl eaeal pA have ek Sane 30 00 253 86

UIA ESAT e s cioe ee ood Sa betes We eaie ren nell ecm 39 02 292 88

BROADEN eB Rod osteo ila ake DPE AR Coe soa 19 12 312 00

BaSEs MRT Pred SE are hd so ehe a enrat eu Aue RU NCne Mary alse chee) 18 06 330 06

USEE YT ORIN 0A Bek el a eg LU Re mana AM 19 32 349 38

RI a hee ae ytd gckigiaies Smita natn ae Talk Tall asia pheloli «Bi 22 89 372) 27

EERO ee Se lus se rale and ON i ate eae se OND EO alae hata 50 77 423 O4

OSPR TET MS WLhNreaiuceca'e i Siar higtw al efale alam tore ie did aia eisie ce ie 45 99 469 03

MMP LON Scone sigh ie eli ac ia stata e arate a Bist etipia ald a yale nace 86 43 555 40

eI er eye NA ss tar Ih it aye aia BE aN URLOAVA LN at nit ace oat ges alae 97 90 653 36

BERR ERNE A, 5 2h Vieaislalmie ela b Maat ME CREP OUR SIRE Aor 102 65 756 OL

SETS SSG 8 INGER ACA a ARE SF a PS WR 388 II 1144 12

Dae et ee ein ain a a aie'a Gs na las Acabatalelanels ins!» 9a 275 12 I4IQ 24

He ya ol kMtors IR le CPA a I ARB alias AN RA a 284 55 1703 79

PARE Te et ere ate Walaa a's «ai nis's/s, 2 vie traiole telat njghele sian! a's 212 80 1916 59

BMV LIN kale tan leit ale: oe oe Solaire eietatae aly alm dis tere 164 50 2081 09

168 PROCEEDINGS OF THE

CONTRIBUTORS TO SPENCER-TOLLES FUND GIVING $50 OR

OVER (CONSTITUTION, ARTICLE VII)

John Aspinwall Robert Brown

Troy Scientific Association

Macnus PFLaum,

Custodian. .

SANDUSKY, Ouro, July 7, 1905.

We, the undersigned committee, hereby certify that we have carefully

examined accounts of the Custodian as given in the foregoing report, have

compared the same with the vouchers, and have found the same to correspond

and to be correct.

W. F. MERcER,

E. P. BuFFET,

Auditing Committee.

CONSTITUTION

ARTICLE [

This Association shall be called the AMERICAN MICROSCOPICAL

Society. Its object shall be the encouragement of microscopical

research.

ARTICLE II

Any person interested in microscopical science may become a

member of the Society upon written application and recommenda-

tion by two members and election by the Executive Committee.

Honorary members may also be elected by the Society on nomina-

tion by the Executive Committee.

ARTICLE III

The officers of this society shall consist of a President and two

Vice-Presidents, who shall hold their office for one year, and shall

be ineligible for re-election for two years after the expiration of

their terms of office, together with a Secretary, a Treasurer, and

a Custodian, who shall each be elected for three years, be eligible

for re-election and whose terms of office shall not be coterminous.

ARTICLE IV

The duties of the officers shall be the same as are usual in similar

organizations ; in addition to which it shall be the duty of the Presi-

dent to deliver an address during the meeting at which he presides ;

of the Custodian to receive and manage the property and permanent

funds of the Society under the direction of the Executive Commit-

tee and in conjunction with a permanent committee to be called the

Spencer-Tolles Fund Committee, and to make a full and specific an-

nual report of the condition of all the property, funds, and effects

in his charge; and of the Secretary to edit and publish the Trans-

actions of the Society.

ARTICLE V

There shall be an Executive Committee, consisting of the officers

of the Society, three members elected by the Society, and the past

Presidents of the Society and of the American Society of Micro-

scopists who still retain membership in this Society.

170 CONSTITUTION AND BY-LAWS

ARTICLE VI

It shall be the duty of the Executive Committee to fix the time and

place of meeting and manage the general affairs of the Society.

ARTICLE VII

The initiation fee shall be $3, and the dues shall be $2 annually,

payable in advance. But any person duly elected may upon payment

of $50 at one time, or in instalments within the same year, become

a life member entitled to all the privileges of membership, but ex-

empt from further dues and fees. All life membership fees shall

become part of the Spencer-Tolles Fund, but during the life of such

member his dues shall be paid out of the income of said fund. A

list of all life-members and of all persons or bodies who have made

donations to the Spencer-Tolles Fund in sums of $50 or over, shall

be printed in every issue of the Transactions. ‘The income of said

fund shall be used exclusively for the encouragement and support of

original investigations within the scope and purpose of this Society.

The principal of the fund shall be kept inviolate.

ArTICLE VIII

The election of officers shall be by ballot.

ARTICLE IX

Amendments to the Constitution may be made by a two-thirds

vote of all members present at any annual meeting, after having

been proposed at the preceding annual meeting.

BY-LAWS

ARTICLE [I

The Executive Committee shall, before the close of the annual

meeting for which they are elected, examine the papers presented

and decide upon their publication or otherwise dispose of them.

All papers accepted for publication must be completed by the

authors and placed in the hands of the Secretary by October Ist

succeeding the meeting.

ArTICLE II

The Secretary shall edit and publish the papers accepted, with the

necessary illustrations.

CONSTITUTION AND BY-LAWS I71

ARTICLE III

The number of copies of Transactions of any meeting shall be

decided at that meeting. But if not decided, the Secretary shall,

unless otherwise ordered by the Executive Committee, print the

same number as for the preceding year.

ARTICLE IV

No applicant shall be considered a member until he has paid his

dues. Any member failing to pay his dues for two consecutive

years, and after two written notifications from the Treasurer, shall

be dropped from the roll, with the privilege of reinstatement at any

time on payment of all arrears. The Transactions shall not be sent

to any member whose dues are unpaid.

ARTICLE V

The election of officers shall be held on the morning of the last

day of the annual meeting. Their terms of office shall commence at

the close of the meeting at which they are elected, and shall con-

tinue until their successors are elected and qualified.

ARTICLE VI

Candidates for office shall be nominated by a committee of five

members of the Society. This committee shall be elected by a

plurality vote, by ballot, after free nomination, on the second day

of the annual meeting.

ARTICLE VII

All motions or resolutions relating to the business of the Society

shall be referred for consideration to the Executive Committee before

discussion and final action by the Society.

ArTIcLE VIII

‘Members of this Society shall have the privilege of enrolling mem-

bers of their families (except men over twenty-one years of age)

for any meeting upon payment of one-half the annual subscrip-

tion ($1).

ARTICLE IX

There shall be a standing committee known as the Spencer-Tolles

Fund Committee to take general charge of the fund and to recom-

mend annually what part of the income shall be expended for the

172 CONSTITUTION AND BY-LAWS

encouragement of research, but the apportionment of the sum thus

set apart shall be made by the Executive Committee.

The Spencer-Tolles Fund Committee shall also have general

charge of the expenditure of such money as may be apportioned,

under the conditions laid down by the Society for its use.

The Custodian shall be an ex-officio member of this committee.

ARTICLE X

The Executive Committee shall have the power annually to ap-

point two members to represent the Society on the Council of the

American Association for the Advancement of Science, in accord-

ance with the regulations of the latter organization.

Revised by the Society, July, 1903.

LIST OF MEMBERS

(Corrected to December 1, 1906)

HONORARY MEMBERS

Crisp, FranK, LL.B., B.A., F.R.MLS.,

5 Landsdowne Road, Notting Hill, London, England

DALLINGER, Rev. W. H., F.R.S., F.R.M.S.,

Ingleside, Lee, S. E., London, England

TIVATIO ..69 Burling Lane, New Rochelle, N. Y.

Warp, R. ite ke M., ‘MD. FR. Deeb ety st 53 Fourth St., Troy, N. Y.

LIFE MEMBERS

Pee Tee ST AVON sd ary a eich a wim Saas ee Os ele 489 Fifth Ave., New York City

BIR WE IT AAS OBERT <5 s*bcrs 6b dleieied, ve tam ee es Observatory Place, New Haven, Conn.

E.viott, Pror. ArTHUR H...................4 Irving Place, New York City

MRL TOTIN Cactcae Sil an aps vie fA de te aot Chicago Beach Hotel, Chicago, IIl.

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 ELECTED DURING THE YEAR 1905-6

(For addresses see regular list)

ARNOLD, SETH F. Hyatt, J. D.

Brown, J. STANFORD (Honorary Member)

(Life Member) LANE, MICHAEL A.

Do.tspy, Epwarp P. Porter, ALBERT B.

Etxtiotr, ArTHUR H. Rocers, B. F., M.D.

(Life Member) SwINcLe, Leroy D.

HATELY, JOHN C. WELLMAN, F. C., M.D.

(Life Member) WricHT, SYDNEY A.

HEALD, FrepertcK D., Ph.D.

ALLEGER, WALTER W., M.D., ’94......---143 U St., N. W., Washington, D. C.

ALLEN, WYNFRED E., A.M, ’04......-0.020200. State Normal, Kearney, Neb.

UROL eres OE 120, ase 5)‘ s slic an bain S 155 W. Concord St., Boston, Mass.

ANSPIN WALL, JOBNe: Nis, (MCE), 700). sue aise Sean swe deed wees Newburgh, N. Y.

POOOD Fost St PO Cea eres a ein weseieik aime Highlands P. O., Monmouth Co., N. J.

Atwoop, H. F., 78........+++++++e+++-16 Seneca Parkway, Rochester, N. Y.

174 _ AMERICAN MICROSCOPICAL SOCIETY

BARCLAY) LOUIS Pic UM Des (OS. cane vale mine oe 537 St. Paul St., Rochester, N. Y.

BARKER, ALBERT S., ’07..........1033 Witherspoon Bldg., Philadelphia, Pa.

BARKER, FRANKLIN D., A.M., ’03...... University of Nebraska, Lincoln, Neb.

BAUSCH, EDWARD, Fos vale Gu viek Hewie ob whe 179 N. St. Paul St., Rochester, N. Y.

BAUSCH, HENRY, "86.00 gece cs cee sa vves 0735 Ne ot. Paul st, Rochester, Now.

BauscH, WILLIAM, ’88..... Shile ee ales 4 ek VU e eC OCIES LET. WIN cate

BEAL, Pror. JAMES HARTLEY, 266, SLB IC SPD bey EAN A) Scio College, Scio, Ohio.

BEARDSLEY, | PROF, WAGE Crea culiceh Dewidiak okie’ 1412 Tenth St., Greeley, Colo.

BELL, ALBERT T., B.S., A.M., ’03..Neb. Wesleyan Univ., University Place, Neb.

Bert) CLARK) WS: FLU vase an os aisle gres 39 Broadway, New York City

BENNETT, HEnry C., ’93......Hotel Longacre, 157 W. 47th St., New York City

BENSLEY, B. A., Ph.D. (Col.), ’05..Biol. Dept., Uni. of Toronto, Toronto, Can.

PERING Ji EDWARD OOLAN Wa bk dem he sae ees 421 W. William St., Decatur, Ill.

BEsSEY, ProF, CHARLES Epwin, Ph.D., LL.D., ’98............... Lincoln, Neb.

BEYER, Pror. Geo. E., ’99..............Tulane University, New Orleans, La.

Birce, Pror. E. A., Sc.D., UL.D., ’99...... Univ. of Wisconsin, Madison, Wis.

BiscoE, Pror. Tuomas D., ’ol...............-404 Front St., Marietta, Ohio

BLeILeE, A. M., M.D., ’81............Ohio State University, Columbus, Ohio

BopINnE, Pror. DoNALDSON, ’96........303 W. Main St., Crawfordsville, Ind.

BootH, Mary A., F.R.M.S., ’82........ 60 Dartmouth St., Springfield, Mass.

Bover, C. S., A.M., ’92. 2... 0000600000. 3223 Clifford’ St, ' Philadelphia, \Pa:

Beri; (GRO PF OOu cals cidves auane ew tea kk beninel ee ka Reais Sistersville, W. Va.

BROMLEY, Rosert Innis, M. D., a Appr maratapaie payline ergs Mews. £ Sonora, Cal.

Brooxkover, CHas., A.B., M.S., wie -Buchtel Coll., Akron, Ohio

Brown, N. How ann, ’gr. pilots or Chestnut ote Phitadel penal Pa.

BrunpacE, A. H., M.D., aid vied by ures Bushwick Ave., Brooklyn, N. Y.

SUP RET Par Lae OG us uk law s beaten aun se Weare 108 Fulton St., N. Y. City

Bult, JAMES Enaar, Esq., ’92.. .seeeee+.-I41 Broadway, New York City

Burritt, Pror. T. J., Ph.D., 8 ..Urbana, III.

Burt, Pror. Epwarp A., Ph. D., hits) Niadlebuey ‘caine Middlebury Vt.

Busu, Miss Bertua E., M.D., 498 PN a ae aie eae 808 Morse Ave., Chicago, IIl.

TRS). Oe eo ee etn ne a ttyonh ange 114 W. Second St., Oil City, Pa.

CALDWELL, Otis W., Ph.D., ’03................State Normal, Charleston, Il.

CARPENTER, THOS. B., M.D., ’99.............-533 Franklin St., Buffalo, N. Y.

CarRTER, JOHN E., ’86..Knox and Coulter Sts., Germantown, Philadelphia, Pa.

CLARK, GAYLORD P., M.D., ’96.......... 619 W. Genesee St., Syracuse, N. Y.

CiarK, GrorcE Epw., M.D., ’96. .Genesee St., Skaneateles, Onondaga Co., N. Y..

CLEMENTS, Freperic E., A.M., Ph.D., ’98....Univ. of Nebraska, Lincoln, Neb.

CLEMENTS, Mrs. Epira ScHwartz, A.M., Ph.D., ’o3,

University of Nebraska, Lincoln, Neb,

COATS: Aico 1s) static nace a tai ices cian on ine a Central Y. M. C. A., Chicago, Ill.

Cocks, Pror. REGINALD S., ’99....McDonogh High School, New Orleans, La.

CorFin, RoBERT, ’00.............-R. F. D. 2, Bedford City, Bedford Co., Va.

Cotton, A. I., M.D., ’o4.......:.:......Ferry and Otis Place, Buffalo, N. Y.

Crosny, (Gyes HRT ALB OS ol ia miele areas teab en 43 East Ave., Ithaca, N. Y.

Coucu, Francis G., ’86,

Kalish Pharmacy, 100 E. Twenty-third St., New York City

LIST OF MEMBERS 175

Cox, Cuas. F., F.R.S.M., ’85........Grand Central Station, New York City

Craic, THOMAS, ’03..............597 Sherbrooke St., W., Montreal, Canada

ERANDALES CrE0. C4) Boos, MDE vas ya) eee 4287 Olive St., St. Louis, Mo.

SRR LLL. OO. ss wsadine eae eal ttn’ 209 Locust St., Evansville, Ind.

DisBrow, WILtiAM S., M.D., Ph.G., ’or........ 151 Orchard St., Newark, N. J.

PO EDWARD Pc OO... ba Uedaae o's 6 ois 4 tac e's 948 N. 43d St., Philadelphia, Pa.

Dorp, Wo. H., ’o4. . Seed okies «20% Clampshire St. putaien avait.

Dorr, S. Hopart, Ph. G, 19s. Rea taa tbe tone t 907 Seventh St., Buffalo, N. Y.

OO RA TEL DAES 6p MeN OP Sol tls a DD se De ER Box 1033, Rochester, N. Y.

DUNCANSON, Pror. TAENBYS Bes) By fen OSs os State Normal, Peru, Neb.

Par PE RVR) FOETENO Vir Df) 00d shite Werte ac Vigl tid died o's ek San José, Costa Rica

Epcar, Tuos. O., B.S., ’04.. FSA Ne .123 E. 25th St., Chicago, Ill.

EIGENMANN, Pror. C. H., 95. ; +650 Rinne Ave., Bloomington, Ind.

ELuiotr, LuTHER B., ’o08. . es .17 Birr St., Rochester, N. Y.

Exrop, Pror. Morton J., M.A. MS. 108,

University of Montana, Missoula, Mont.

Eusner, JoHNn, M.D., ’83.............+.---1014 Fourteenth St., Denver, Colo.

(OMS TS a a ahh Re 8 OR A es ee a aE Ra 16 Pearl St., Council Bluffs, Iowa

EweE.t, MarsHALt D., M.D., LL.D., ’85............59 Clark St., Chicago, Ill.

Eyre, Joun W. H., M.D., M.S., F.R.M.S., ’o9,

Guy’s Hospital, London, E. C., England

PREPTOU DOL PTI). 28 toc so ok wie 5 vie < elem ed 520 E. Main St., Columbus, Ohio

ett. Geo I, M.D, FORMS, ’78... 02685) Miller Blk., North Yakima, Wash.

FELLows, Cuas. S., F.R.M.S., ’83,

g12 Chamber of Commerce, Minneapolis, Minn.

SORECUISOM VEBADE D1 .o., 5 FILLES 09254 2 cabas a's orn a ve ne sigie'aintl wait Blacksburg, Va.

Ferris, Pror. Harry B., M.D., ’96.......... 118 York St., New Haven, Conn.

SPEDE Rs AW My | Boy Dass OG sien ve ae poe e aie aviclsie 2 Union Place, Troy, N. Y.

Fiscu, Cart, M.D., Ph.D., 03 Bes ie Rikon wets 6 3212 Pine St., St. Louis, Mo.

FiscHEr, ALF., ’02. seccescccscccesesee. 046 Broadway, Milwaukee, Wis.

Fiscuer, Cuas. E. M., iets LIE Pare ae | Spree CPN 259 S. Clinton St., Chicago, Ill.

FiscHER, MAX, ’93.. ; ..Zeiss Optical Works, Jena, Germany

FISHER, REv. Seinen S., “ScD., DD, ith Ae eters ce West Lafayette, Ohio

Fuint, James M., M.D., ’or. ..Stoneleigh Court, Washington, D. C.

BO ei MDs, OLS aes ek nes ney 202 S. Thirty-first Ave., Omaha, Neb.

Forpyce, Cuares, B.S., A.M., Ph.D., ’08,

Nebraska A as University, University Place, Neb.

Foster, EDWARD, ’99... ..The Daily Picayune, New Orleans, La.

FuLier, CHAs. G., MD. ERMS, 81 AES oh Reliance Bldg., Chicago, Ill.

Futon, Harry R., A.B., . La. Agric. Exp. Sta., Baton Rouge, La.

Furniss, H. W., M.D., fed vont EO Oke eb Se Gonshlate.. Gantacesra ar

Gace, Pror. Stmon H., B.S., ’82.........-..-Cornell University, Ithaca, N. Y.

Gace, Mrs. SUSANNA PHELPS, ’87.....-+-0eee00e 4 South Ave., Ithaca, N. Y.

GaLLoway, Pror. T. W., ’or..........James Millikin University, Decatur, Ill.

CATES PL MEB SOO ii wise vic pines vlneic viele atinleg males eulbgyaln ants Chevy Chase, Md.

176 AMERICAN MICROSCOPICAL SOCIETY

MAEETY, JOHN, D1) "OD. ep itve sso scutes VENT ORNS Eee . Sparta, Kent Co., Mich.

Gittmore, Miss GERTRUDE A., B.A., ’03......27 Charlotte Ave., Detroit, Mich.

RRA TAC OR. an Ueldbth dente adel varied 424 E. Fourteenth St., Oakland, Cal.

GrosskKopF, Ernest C., M.D., ’90.........2.00. 187 36th St., Milwaukee, Wis.

SEAAG?. DD. FE, MIU OOrissne ve ee ceectinh wake eb Nin abet’ Liberty Center, Ohio

BIRT VICTOR: 5 COLD aclan 409 oiaiee sien telens 191t Webster St., San Francisco, Cal.

HANAMAN, C. E., F.R.MLS., ’79.......2.. State and Second Sts., Troy, N. Y.

HANKINSON, T. L., ’03.........0.0+e+e00+e++-e9tate Normal, Charleston, Ill.

LIATFIELD, JOHW J.) Diy CAV abled Leesa sed HOLS eben Malott Park, Ind.

Pktp: FE: DL PRS POO. isu aces een boveds te Univ. of Nebraska, Lincoln, Neb.

HERTZLER, ARTHUR E., M.D., ’96......... 508 Altman Bldg., Kansas City, Mo.

Hertzoc, MAximiLian, M.D., ’or,

Room 800, 103 E. Randolph St., Chicago, III.

His, Hiseeeero MPa err en oo Lo ea ely 24 High St., Buffalo, N. Y.

Hixtton, Davin Ciark, A.M., M.D., ’OI......... e000: 1240 O St., Lincoln, Neb.

TOOK AE L208 iL ida abe dalek Oice ces as td beah pa eee Jefferson City, Mo.

HorrMANn, Jos. H., M.D., ’96.. i .111 Steuben St., Pittsburg, Pa.

Ho tis, Freperick S., Ph.D., ‘99. . ) Wale Medical School, New Haven, Conn.

FIOLMRS,; PAS sg a OR wack csseakk cease 205 Jackson Block, Denver, Colo.

Hoskins, WM., ’79.. .-Room 54, 81 S. Clark St., Chicago, III.

How.anp, HEnry R., Ta yee 8... . .367 Seventh St., Buffalo, N. Y.

Humpurey, Pror. O. D., Ph.D., 195. eas State Normal School, Jamaica, N. Y.

Ives, Freperic E., ’o2.........Woodcliff-on-Hudson, Weehawken P. O., N. J.

Jackson, Daniet Dana, B.S., ’99........941 President St., Brooklyn, N. Y.

JAMES, FrankK L., Ph.D., M.D., ’82......... 514 Century Bldg., St. Louis, Mo.

Jounson, Frank S., M.D., F.R.MLS., ’93...... 2521 Prairie Ave., Chicago, IIl.

Jounson, Wo. D., M.D., ‘08. PPL Seperate Ay ec SL h, Batavia, N. Y.

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KRAFFT, Worse a SA Bie AAR EARS 3 411 W. Fifty-ninth St,, New York City

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Lanpacre, F. L., B.A., ’03.. ...+.-Ohio State University, Columbus, Ohio

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INDEX.

Acetabularia, 62

Alsire lakes, of Pike’s Peak region,

biology of, 75 et seq.

Alternation of hosts in life history of

parasites, II

Animals, as breeders of disease-

germs, 9; as carriers of disease-

germs, 6; as preventers of disease,

19; as producers of disease, 13; re-

lation of, to disease, 5

Announcement by the Secretary, 157

Anodyomene, 59

Apparatus, I5I

Avrainvillea, 55

Bacteria, phylogeny of, 63; relation

to Flagellata, 64; relation to Cyano-

phyceae, 65

Barker, F. D., Variations in the Vitel-

laria and Vitelline Ducts of three

Distomes of the Genus Opis-

thorchis, 99

Bausch and Lomb, New Model Pro-

jection Apparatus, Lantern “D,”

153

Bessey, Charles E., The Structure and

Classification of the Siphonales,

etc., 47

Biological Study of the Lakes of the

Pike’s Peak region, A,—Prelim-

inary Report, by H. L. Shantz, 75

Boodlea, 59

Book Review, 154

Botrydiaceae, characters, 56; key to

genera, 57

Botrydium, 57

Botryophora, 61

By-Laws, 170

Caulerpaceae, characters, 61

Caulerpa, 61

Chaetomorpha, 53

Chamaedoris, 59

Cladophoraceae, characters and key to

genera, 52

Cladophora, 53

Codiaceae, characters and key to

genera, 54

Codiolum, 57

Codium, 55

Colonies, of Volvox, finer structure

of, 139; number of cells in, 138;

reproductive cells in, 134

Colorado. Biological study of lakes

of Pike’s Peak region, 75

Constitution, 169

Contribution to the Microscopic

Study of Pen and Ink Lines, A, by

Marshall D. Ewell, 21

Custodian, report of, Magnus Pflaum,

167

Cvanophyceae, relation of bacteria to,

65

Cymopolia, 62

Cystodictyon, 59

Dasycladaceae, characters and key to

genera, 61

Dictyosphaeria, 59

Disease, the relation of animals to, 5

Disease germs, carried externally by

animals, 6; carried internally by

animals, 8

Dithyridium cynocephali n. sp., 32

Dolby, E. P., The Pietzsch Micro-

tome, 152

Ducts, vitelline, of Opisthorchis spp.,

99

Ewell, Marshall D., A Contribution

to the Microscopic Study of Pen

and Ink Lines, 21

181

182

Fauna, of lakes of Pike’s Peak re-

gion, 85, 92

Filipino, Porocephalus constrictus in

a native, 41

Flagellata, relation of bacteria to, 64

Flora, of lakes of Pike’s Peak region,

84, 92

Frog, trypanosomes in the blood of,

25

Gomontia, 53

Halimeda, 55

Hare, Chas. B., with Herzog, Maxi-

milian, Porocephalus constrictus in

a native Filipino, 41

Heald, F. D., The Phylogeny of Bac-

teria, 63

Herzog, Maximilian, and Hare, Chas.

B., Porocephalus constrictus in a

native Filipino, 41

Horses, a nematode of, heretofore un-

reported from the United States,

33

Lakes, of Pike’s Peak region, biolog-

ical study of, 75

Legal microscopy. Microscopic study

of pen and ink lines, 21

Lines, pen and ink, microscopic study

of, 21

Linguatulid, Porocephalus constric-

tus, in a native Filipino, 41

List, of members, 173; of officers, 3;

of subscribers, 180

Liver, linguatulid parasite in human,

4I

Meeting, twenty-eighth, minutes of,

159

Members, list of, 173

Microdictyon, 59

Microscopy of Vegetable Foods, The,

by A. L. Winton, review, 154

Microtome, Pietzsch, 152

Minutes of the twenty-eighth annual

meeting, 159

Muscles, of marsupial wolf, tape-

worm cysts in, 31

INDEX. |

Nebraska. New forms of Volvorx

from Crete, Nebr., 123

Nematode, of horses, heretofore un-

reported from the United States,

33

Neomeris, 62

New apparatus, Reviews, etc., 151

New Forms of Volvox, by J. H.

Powers, 123

Occurrence of Trypanosomes in the

Blood of Rana clamata, On the, by

James H. Stebbins, Jr., 25

Officers, list of, 3

Opisthorchis felineus, Riv., vitellaria

and vitelline ducts, 106

Opisthorchis lancea Dies., vitellaria

and vitelline ducts, 107

Opisthorchis pseudofelineus Ward,

vitellaria and vitelline ducts, 99

Parasites, alternation of hosts in, II;

as producers of disease, 16; effects

of, 14; in relation to diseases, 9 et

seq.; linguatulid, in native Filipino,

41; nematode, of horses, 33; tape-

worm, in muscles of a marsupial

wolf, 31; trypanosome, in blood of

frog, 25; trypanosome, in rat, III

Penicillus 55

Pflaum, Magnus, Report of the Cus-

todian, 167

Photomicrographic Outfit, A, by J.

H. Stebbins, Jr., 151

Phyllosiphon, 54

Phyllosiphonaceae, characters, 54

Phylogeny of Bacteria, The, by F.

D. Heald, 63

Pietzsch Microtome, The, by E. P.

Dolby, 152

Pike’s Peak region, biological study

of lakes of, 75

Pithophora, 53

Plains lakes, of Pike’s Peak region,

biology of, 81 et seq.

Plankton, of lakes of Pike’s Peak

region, 85

Porocephalus armillatus Wyman, 44

INDEX.

Porocephalus constrictus von Siebold,

4I

Porocephalus constrictus in a native

Filipino, by Maximilian Herzog

and Chas. B. Hare, 41

Powers, J. H., New Forms of Vol-

VOX, 123

President, Annual Address of, by

Henry B. Ward, 5

Probstmayria nov. gen., 35; P. vivi-

para (Probstmayr), 35 ify

Probstmayria vivipara (Probstmayr,

1865) Ransom, 1907, a Nematode of

Horses heretofore unreported from

the United States, by B. H. Ran-

som, 33

Projection apparatus, new model,

Bausch and Lomb, Lantern “D,”

153 .

Protosiphon, 57

Protozoa, as producers of disease, 17

Ransom, B. H., Tapeworm Cysts

(Dithyridium cynocephali n. sp.) in

the muscles of a Marsupial Wolf~

(Thylacinus cynocephalus), 31;

Probstmayria vivipara (Probst-

mayr, 1865) Ransom, 1907, a Nema-

tode of Horses heretofore unre-

ported from the United States, 33

Rat, trypanosome of, III

Relation of Animals to Disease, The,

by Henry B. Ward, 5

Report of the Custodian,

Pflaum, 167

Report of the Treasurer, J. C. Smith,

166

Reproductive colonies, in Volvox, 134

Reviews, 154

Rhipocephalus, 55

Rhizoclonium, 53

Ross Microscopes, 156

Magnus

Secretary, announcement by the, 157

Shantz, H. L., A Biological Study

of the Lakes of the Pike’s Peak

Region—Preliminary Report, 75

Siphonales, characters and key to

183

families, 51; structure and clasifica-

tion of, 47

Siphonocladus, 59

Smith, J. C., Report of the Treasurer,

166

Some Studies on Trypanosoma

lewisi, by Leroy D. Swingle, 111

Spencer-Tolles Fund, report of, 167

Sperm development, in Volvox, 124

Sphaeropleaceae, characters, 53

Sphaeroplea, 54

Stebbins, James H., Jr., A Photo-

micrographic Outfit, 151; On the

Occurrence of Trypanosomes in

the Blood of Rana clamata, 25

Structure and Classification of the

Siphonales, The, with a Rearrange-

ment of the Principal North

America Genera, by Charles E.

Bessey, 47

Struvea, 59

Subscribers, list of, 180

Swingle, Leroy D., Some Studies on

Trypanosoma lewisi, 111

Tapeworm, cysts in muscles of a

marsupial wolf, 31

Tapeworm Cysts (Dithyridium cyno-

cephali n. sp.) in the Muscles of a

Marsupial Wolf (Thylacinus Cyno-

cephalus), by B. H. Ransom, 31

Treasurer, report of, by J. C. Smith,

166

Trypanosoma clamatae n.

multiplication of, 27

Trypanosome lewisi, 111; agglomera-

tion, 116; morphology of, 112, 118;

multiplication of, 113; transmission

of, 117

Trypanosomes, in the blood of Rana

clamata, 25; multiplication of, 27

Spe}

Udotea, 55

Urospora, 52

Valonia, 58

Valoniaceae, characters and key to

genera, 58

184

Variations in the Vitellaria and Vitel-

line Ducts of three Distomes of the

Genus Opisthorchis, by F. D.

Barker, 99

Vaucheria, 56

Vaucheriaceae, characters, 56

Vitellaria, of Opisthorchis spp., 99

Vitelline ducts, of Opisthorchis spp.,

99

Volvox, new forms of, 123; number

of cells in, 138; reproductive colo-

INDEX.

nies in, 134; sperm development in,

124

Ward, Henry B., Annual Address of

the President—The Relation of

Animals to Disease, 5 .

Watson and Sons’ Instruments, 155

Winton, A. L., The Microscopy of

Vegetable Foods, review, 154

Wolf, marsupial (Thylacinus cynoce-

phalus), tapeworm cysts in muscles

of, 31

THE AMERICAN MICROSCOPICAL SOCIETY I

SPENCER

MICROSCOPES

EXCEL IN OPTICAL PERFECTION,

BEAUTY AND PRACTICAL UTILITY

EACH OF | WO STYLES or

FINE ADJUSTMENTS

IS THE

PROTECTED “™

BY A HANDLE

PROVIDED FOR CARRYING

THE MICROSCOPE

THE STAGES ARE THE LARGEST, ARE COMPLETELY COVERED

WITH VULCANITE; AND THE SUBSTAGE EQUIPMENT

1S THE MOST CONVENIENT YET PRODUCED

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OF MICROSCOPES AND ACCESSORIES

SPENCER LENS CO., BUFFALO, N. Y.

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II THE AMERICAN MICROSCOPICAL SOCIETY

THE KNY-SCHEERER CO.

AND THE

LABORATORY AND SCHOOL SUPPLY CO.

in combination with

The United Factories of Laboratory Supplies

Vereinigte Fabriken fur Laboratoriumsbedart

Proprietors

MAX KAEHLER & MARTINI—DRS. PETERS & ROST

BERLIN N.

One of the world’s largest manufacturers of

Chemical, Physical and Biological Apparatus

are prepared to bill either import orders ot orders from New York stock to the

satisfaction of the customer who desires

A SUPERIOR QUALITY OF GOODS

Also: Anatomical Models, Osteological, Biological and Anatomical

Preparations, N atural History Specimens, Wall Charts, etc.

For up-to-date new catalogue address

THE KNY-SCHEERER CO.

225 to 233 Fourth Ave.

Department of Natural Science and Laboratory Supplies New York City

James H. Stebbins, Jr., S.B., M.S., Ph.D.

CHEMIST AND MICROSCOPIST

Cee eee ee —__—_______ ee

3 West 2gth St., New York

Chemical and bacteriological examination of water,

milk, and all matters relating to .public health.

Microscopic examination of papers, fibers, starches,

food products, drugs, metals, etc.

Photomicrographs, and Lantern Slides made to order

at short notice

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THE AMERICAN MICROSCOPICAL SOCIETY

Microscopes

for Physicians and all Scientific Purposes

Photo-micrographic

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Visible and

Ultraviolet Light.

©utfits for the

Investigation of

Ultramicroscopical

Particles.

Epidiascope and

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——— — Write for

i Cc. LEISS ‘6 99

C—————— . Catalogue “MA.

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*London., Vienna. St. Petersburg.

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IV THE AMERICAN MICROSCOPICAL SOCIETY

BG Medals and Highest Awards at all the Great International Exhibitions “@%

© ROSS? “stanparo”

{\\ MICROSCOPES,

SPECIALLY DESIGNED aft, THE Usk OF

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serviceable English In-

struments ever offered at

such moderate prices.”’

PRICE LISTS OF

ss Microscopes,

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: = POST FREE. = ==

"PHOTOGRAPHIC OUTFITS, ETC.

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(esTaBLISHED 1830) GLAND.

BIOLOGISTS, MICROSCOPISTS

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——— IN ALL BRANCHES

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- THE AMERICAN MICROSCOPICAL SOCIETY V

Bausch & Lomb

Microscopes

are used in the majority of college

laboratories because they are the

best, optically and mechanically.

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New York Boston Chicago San Francisco

Frankfurt a/M. Germany.

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VI THE AMERICAN MICROSCOPICAL SOCIETY.

EDWARD PENNOCK

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=. OF.THE LEADING (Ye SSS

Sole American

se rer” “Tallquist Hemoglobin Scale

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Makersace Microscopes and Objectives

Have constructed a New Semi-Apochromatic Homogeneous Im-

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long or short tube—a fine lens for bacteriological work. The

glass used in its construction is free from deterioration. Price, £5.

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New dry Apochromatic Condenser corrected for thickness of

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THE AMERICAN MICROSCOPICAL SOCIETY VIl

LEITZ’S

New Microscopes

With most improved form of

Side Fine Adjustment, Extra Large

Stage, Dust Proof Nosepiece

We carry in stock a complete line of

Microscopes and Accessories

Projection Apparatus

for episcopic, microscopic and lantern slide projection

‘ Microtomes and Knives

Bacteriological Apparatus, Laboratory Supplies

Dr. Grubler’s Stains

east wnst. ERNST LEITZ ¢sscer

)

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