“ aves fe Z baal we . a - ~ : m ; “ : eet hs veal " 2 Pe ae personae ~ ~o 8 9 Pa srr yay opr LYFE Te ie seh rn ny ee da f icreiiian ek vi avis c v4 hoe tah OFA " , see, ‘i Uo ba 5 ut an 4 ey, . su bey Uy } 3 4 “ Bo ee ¥ Pay rs Ter] are \ : ins ie N ' tiv j wy Ake yp ae rs ve! 4 ube >. bey Fe 4 (Ay: ‘ADS ‘ n ' a ; , > ~ i inn i yi i a ft Oia | Ai ( ny ane) Ly & ») ie iy Pat re aA A Da NY ph) ae i) f ‘ i Be | eas A) PIT AT j < ne wes iy! ij 14 Wy ala l es vin) ra} 1 Five ] Tin | | Mat, eA aA wine) he bia, L # 8.) A ry / f 7: ‘ \ | ; f ie ry Ait / iA tr aL 4 / a we 1s aa : Ae , —_ == = 7 oi j Ail j v 1 ie | r Ty ls) I , rule a ny pir ; rh I ih a yy it oie hapen woe i? yap A, i Neti i nL f ! ‘ i i I t ‘ Ah ) ‘ Yh fi f } x 4 4 " : i , i ( ‘ i = V j i y Pare scu| \ al Pye y A j ' (i f ’ a i in j i j toh) | { , \ i {| Ve i Lae mee) : t(j ' | ‘id TRANSACTIONS OF THE American Microscopica! Society ORGANIZED 1878 INCORPORATED 1891 EDITED BY THE SECRETARY Twenty-Third Annual Meeting NEW YORK CITY, JUNE 28, 29, anv 80, 1900 9 VOLUME XXII LINCOLN, NEB.: STATE JOURNAL COMPANY, PRINTERS 1901 OFFICERS FOR 1900-1901 oe OH President: CH. EIGENMANN...6..00.5000000 Neo sales Bloomington, Ind. Vice Presidents: CHARLES M. VORCE.........4 oh Ol eat Cleveland, Ohio EDWARD PENNOCK.......... ‘ 5 rant Philadelphia, Pa. Secretary: HEenmse B. WARD... 5.0.06. 0 52 ae warar .Lineoln, Neb. TrEQsurers. JorCs SMITHS Ai esic/g las eisreteierel aoe aie ...- New Orleans, La. Custodian: MAGNUS PFLAUM.........-.....- cS Of. >... .Pittsburg, Pa. ELECTIVE MEMBERS OF THE EXECUTIVE COMMITTEE. JOHN AUSTEN WAM ire 2 svajet cera steers srctojsteyctclevate eerie oletehetererscherederete New York City CHARLES VAL TIMOROMD 5 aisle ysieriers alsa) te-ciepe elolekero eke lonvetel st olalleratete Berkeley, Cal. LAGS STEN TSDD So Sh lab eesie oi trae. eis tte iaiias Wiclioiete aceite lotlegereas etereeterale Des Moines, lowa EX-OFFICIO MEMBERS OF EXECUTIVE COMMITTEE Past Presidents still retaining membership in the Society RHC Wer Mey, Chine WES S25 OL MEO yeuNe N.5 at Indianapolis, Ind., 1878, and at Buffalo, N. Y., 1879. H. L. Smiru, LL. D., of Geneva, N. Y., at Detroit, Mich., 1880, and at Cleveland, O., 1885. J. D. Hyatt, of New York City, at Columbus, O., 1881. ALBERT McCALLA, Ph. D., of Fairfield, Ifa., at Chicago, Ill., 1883. T. J. BURRILL, PH. D., of Champaign, I1., at Chautauqua, N. Y., 1886. Gro. E. Brun, M: D.or. R: MAS: of Buitalo, No Y-.; at Detroit, Mich., 1890. Frank L. Jamus, Pu. D., M. D., of St. Louis, Mo., at Washington, D. C., 1891. MaRsHALL D. EWELL, M. D., of Chicago, M1, at Rochester, N. Y., 1892. Simon Henry GAGE, B. S., of Ithaca, N. Y., at Ithaca, No Y., 89a: A. CLIFFORD MERCER, M. D., F. R. M. S., of Syracuse, N. Y., at Pittsburg, Pa., 1896. E. W. CLAYPOLE, B. A., D. Se. (Lond.), F. G. S., of Pasadena, Cal., at Toledo, O.,'1897. WC. KRAUSS: MDs EL ROMA S:, of Buitalo, INOY:, at Columbus, O., 1899. A. M. BLEILE, M. D., of Columbus, O., at New York City, 1900. The Society does not hold itself responsible for the opinions expressed by mem- bers in its published Proceedings unless indorsed by a special vote. TABLE OF CONTENTS FOR VOLUME XXII The Annual Address of the President: The Detection and Recog- inibiikovey Nose IBlkoxoxol, Fenway MLB IeWeQea as a racgdaloodcodaunooeuboonodd 1 Some Advantages of Field-Work on Surface Water Supplies, by lEloMeo) ING GERIELCIOG Gadi nodboooddoocHRonCdouddmodesumoedocg ons 13 The Work of the Mt. Prospect Laboratory of the Brooklyn Water Works, by George C. Whipple, with Plates I to IV............ 25 Methods of Producing Enlargements and Lantern Slides of Micro- scopic Objects for Class Demonstrations, by John Aspinwall... 41 On the Distribution of Growths in Surface Water Supplies and on the Method of Collecting Samples for Examination, by Fred- erick: S. Hollis i with) Plates! Vi GOVE ca crayeislaretelerersiaarercieleteisterets 49 Limnological Investigations at Flathead Lake, Montana, and Vi- cinity, July, 1899, by Morton J. Elrod, with Plates IX to XVII 63 An Addition to the Parasites of the Human Ear, by Roscoe Pound. witley Plater RWW esperar eras ieveiet even eval ohare eenenecaisherelare sls orolevets 81 The Modern Conception of the Structure and Classification of Desmids, with a Revision of the Tribes and a Rearrangement of the North American Genera, by Charles E. Bessey, with HEM ENGIO NG hiivetcad Vat chavia otisuaneraienalshele al aveierere sTalel iene =,/a\sre lai evayererey slay stalate aheleys 89 Photo-spectography of Colored Fluids, by Moses C. White, with EATS PRONG yer rolrerciele) lol s)aictszal aeateiotalepohatetetsley ed ailetayeuel ola lelialayathave: avalfat Nob alol eval 99 Description of a New Genus of North American Water Mites, with Observations on the Classification of the Group, by RODEEU Hl. WoOleOtbe with) late eR eis cists sere: cline over eteielel vrenel aver 105 The Cladocera of Nebraska, by Charles Fordyce, with Plates PONG TT ty Op RONG Wc) cy stekeys evover ole buas eialevever ehoyaveuayetevelioiaverasialte) stahavatayayeiuns tanatet aa 119 Notes on the Parasites of the Lake Fish. III. On the Structure of the Copulatory Organs in Wicrophallus nov. gen., by Henry B. Wijsuas ch e wolbl Esl GE) RONG Ua. chavo yetaiaie: ein) aialas sine yaieereiel a°e la er detelatele akatalunsie; ts 175 Description of a New Cave Salamander, Spelerpes stejnegeri, from the Caves of Southwestern Missouri, by Carl H. Eigenmann, ivi bine ea GES SONOMA TMD Septal KE WANN sey ely ee aly 5 fal allaysalelieavayousley diareret 189 Revoriot the Limnological?Commiss1om.s/)5).\). 2/1 ')2)s10)s\\+1)s0e)e.10 = 0) 193 Necrology: Jacob Dolson Cox, with Plate XXIX.......:......... 197 Moses Clark White, with Plate XOXO, - hese. ce 5s 202 MIE U HES Om cine Acme ite LMC CLIO. |. a6, cja 0a lect «biel ensfiels siuieleistnralaliaie sl avavclere 205 PER SASIE STS EUG I Gdstinsy atotesre)s sl elsecalniera tel eters ralestavetellals ov cveuelaial duavies eet aerate taverns 209 Custodian’s Report, Spencer-Tolles Fund....................e0005 210 WOns LubWULOMN a ycrcosetdsieehe spalet ate sboietal che letancevonel eiarsiaie ts lekeracierertra els sorters 211 BES ys La Siiy ciate teste etal SRC Ce eM EV evel aiey sha ie isha Suueey elon ls, syste! ejay si ershwiar si ele costal olan euate 212 ETS ti Ota NUCTMD ETS etait setae ae tetany i slates sietarolielalatsaleratetaitsvel al atedelalareiata 215 MISE OF SWPSCEUDETS ies «cia lee aya sis iai slave oie.sie' tia \sin is) 8)s\e.e'sa\wleiie «armel whetalleli alae) ela 222 Biennial Index stor Volumes oxox lvama: XOX cfa'screie) «leheieveierchatsraverepeliere 223 ANCG INGE gt USES 0 EYE ON resplendent Aili ot es Ay Ae An Ry RN ea a DY A I _ ERRATUM ; i) \ Page 205, second paragraph, first line, for “Limne Committee” read “Limnological Commission.” TRANSACTIONS OF The American Microscopical Society TWENTY-THIRD ANNUAL MEETING, HELD AT NEW YORK CITY, NEW YORK, JUNE 28, 29, anp 30, 1900 THE ANNUAL ADDRESS OF THE PRESIDENT THE DETECTION AND RECOGNITION OF BLOOD By A, M. BLEILE Following in the path marked out by the steps of some of my predecessors, I have chosen for the subject of this address a theme which is not rigidly microscopical in all of its aspects; nor do I present it as based entirely or even for the largest part on own work or on original observation. Again, the treatment is not rigidly technical, since in a membership made up of persons engaged in very different lines of work, as is the case in this Society, such treatment could at best be of interest to only a very limited number. An attempt has been made to give a resume of a given question with here and there a state- ment based on own experience, and thus it is hoped that a somewhat wider interest, sufficient to hold during its presenta- tion, may be evoked in the subject chosen, which is “The De- tection and Recognition of Blood.” By the detection of blood I mean the finding of blood in a given object, a fluid or a stain; by recognition of blood is meant, in addition to the foregoing, the identification of the find as having come from a particular species of animal and from no other. 1 bo A. M. BLEILE In the detection and recognition of blood are presented two questions of supreme importance and interest in many cases to the physiologist, the physician, and the jurist. Often, too, the layman has in these questions,—even when not personally involved or where his own body or welfare are not, directly concerned, as in medico-legal cases,—at least a curiosity, if not an actual concern, born of that value we all set on affairs affecting the general public weal or woe. While the physiol- ogist is often enough called upon to determine the presence or absence of blood, still in his cases the amount available is in most instances large enough and the specimen fresh enough so that no unusual difficulties present themselves to him in the application and the interpretation of the recognized tests. The physician who wishes to determine the presence or absence of blood in the various exudates and secretions of the body, and on which he will form his diagnosis and even the treat- ment of a case,—two factors on which the whole future pro- gress of the individual may depend,—is apt to meet with diffi- culties more or less great. Sometimes the amount obtainable for examination is quite small; nearly always the blood is mixed with organic fluids of greater or lesser complexity whose own composition may offer obstacles to the use of the otherwise ready and sure tests, only to be overcome by ingenu- ity displayed in meeting the different complications as they ‘arise in individual cases. From a juridic point, however, these questions present a most important aspect and often great complications. In such cases is there many times the most communal interest. Matters of the most vital concern may be at stake in a single case, with a single small fragment at hand for examination, and here the observer meets with his greatest difficulties, not only in carrying on his work so that the results may be certain and satisfactory to himself, but also in carrying it on so that its outcome may be convincing to judge or jury. The problems presented for solution in such cases are two-fold. In many instances it may be sufficient to determine only whether or not blood be present in a given fluid or on a given object in the form of stains or spots, while sometimes there is added to this the desirability or need to recognize the blood as having come from man or a lower animal, and this is by far the more difficult and uncertain— under circumstances impossible—part of the riddle given. The way to its solution is beset with snares and pitfalls to be THE PRESIDENT’S ADDRESS 3 avoided only by most conservative and circumspect considera- tion of all difficulties and sources of error. As to the means at our command for the answer of the ques- tions propounded, you know that we have only two elements peculiar to the blood as contrasted with the contents of the other fluids found in the animal body. These are, on the one hand, the red blood-corpuscles, and on the other their unique constituent, the hemoglobin or oxyhemoglobin; and only the first of these, with present knowledge—the corpuscles, can come into play when it is attempted to recognize the blood as having come from a particular kind of animal. A question of such vital import in its answer has of course received much attention, and among the workers applying themselves to its solution we count several members of this Society. As aids here we have the well-known fact that the red corpuscles in man and all animals (except the Camelidae) are biconcave, non- nucleated, circular disks, and that in birds, reptiles, or fishes, they are biconcave, nucleated, oval (except in cylostoma). This, however, gives a distinction too broad to be of use in many cases, since the question is not often one involving such general differentiation, but narrows itself to the recognition, in nearly every instance, of mammalian bloods, where we have the same general form and where alone differences in size might be invoked to help to the identity of a given specimen. Accordingly the fact being fixed that different kinds of mam- mals do present differences in the sizes of the respective red corpuscles, much effort was given to their measurement in the hope that here might be found the much-desired means of recognition, and work along this line had not a little influence on the perfecting of the optical parts of the microscope and the measuring apparatus employed, since it was felt that only the use of the most exact and accurate appliances could lead to successful or trustworthy conclusions. In the prosecution of this work two ideas, fundamental for its import, seem to have been generally and tacitly accepted by some investiga- tors. The first one was that for every species of mammal there is a fixed size of the corpuscle presenting at most ex- tremely minute variations in different individuals of that species or in the same individual; the second was that in restoring corpuscles from old or dried states the means used were such as guaranteed the re-establishment of the former normal, fresh dimensions. While, then, measurements were 4 A. M. BLEILE being established, and on these men were found enthusiastic enough to declare even under oath that the results were posi- tive and trustworthy, thus taking upon themselves the great- est responsibility in vouching for the correctness of the state- ment that the blood in hand was or was not human blood, newer and better knowledge soon led to a more conservative position. For we know now that differences in size do exist in the corpuscles of one individual and in individuals of the same species, differences so great that they may easily overlap the average measurements given for another species, as for exam- plein dog and man. As to the second premise, that restoration of corpuscles to original volume would be completed by the means employed, few histologists familiar with the delicate nature of these bodies, and having in mind the readiness with which they respond in structure to even slight variations in their surrounding fluid, would be willing to subscribe to this proposition. This method of recognition, then, as between mammalian bloods has been generally given up as untrust- worthy; and while it is easy to distinguish between the oval and nucleated corpuscles of the ovipara and the circular non- nucleated of mammals, it is to the highest degree unsafe ac- cording to more conservative view to attempt more. Success can only be looked for in exceptional cases and under favor- able circumstances, with fresh specimens, where the question is not a general one, but where it is narrowed down to the distinction between two bloods with widely different corpus- cles, i. e., man, or mouse, or squirrel. The finding of the red corpuscles will therefore in nearly every instance mean the detection of hlood, with only a broad statement as to its source; and even in going so far corroborative tests are highly desirable, since the form alone of these bodies is not quite sufficient to establish their identity. In support of this state- ment it may be recalled that certain fungus spores present an appearance almost identical with that of the human red corpuscle, and showing the same dimensions, which have led to error in their interpretation. True, in many cases where such an error was made these bodies were globular and not biconcave, and an inspection by a trained observer would at once have set at rest doubts that might have arisen as to their nature; but in a few instances discoid bodies with an apparent central concayity have been found, thus giving a much closer resemblance to the red corpuscle, demanding a more rigid THE PRESIDENT’S ADDRESS 5 scrutiny for their recognition. That the danger of fallacious finding is a real and not infrequent one is apparent from a perusal of the literature on the subject, in which are given many instances of such mistakes; and one having even a limited experience in this line will be sure to have encoun- tered such bodies in one or two instances. In fact, Richard- son, referring to the various fluids recommended for the ex- traction of corpuscles from old stains, and speaking of one of them—Na,SO,—says, “It must, I think, owe its popularity chiefly to the fact that it contains large quantities of fungus, the spores of which resemble blood corpuscles both in size and appearance and have, I have no doubt, frequently been mistaken for blood-cells.” A very careful study of the chemi- cal composition of blood-cells shows that there are slight dif- ferences in the amounts of alkalies, phosphates, hemoglobin in different animals, but the amount of blood necessary for such determinations is so large as to preclude use of the facts for the purposes before us. Since, then, the answer to be obtained from a study of the corpuscles as such is limited, their available constituent, the hemoglobin, a crystallizable body, has been called upon with the hope of getting from it something definite or trustworthy. This interesting body, also known as the blood-pigment or blood-coloring matter, may therefore be considered in some of its properties, even at the risk of repeating what is everyday knowledge. Hemoglobin occurs in the red cells and belongs chemically to the proteid group, and can be obtained more or less readily in crystals. It is a very unstable body, readily undergoing change and decomposition by agencies inert to other physio- logical constituents; in fact, it owes its physiological value in the organism to the ease with which it may be changed. Heat, weak acids or alkalies will split it up into an albumen—globu- lin—and an iron containing colored organic body, the hematin, and this change will even take place in dried blood when long exposed to the air. It is well known that the form of the crys- tals and the ease with which they may be obtained will differ with the species of the animal from which the blood has come. Accordingly Guelfi made this a basis for some work bearing on the question of the recognition. A 2% NaFl. solution is used with an equal quantity of blood and held at a tempera- ture of 40°, when erystallization will soon take place. Thus there are procured from guinea pig blood tetra-hedra; from 6 A. M. BLEILE the dog’s, prisms; while some other bloods will give needles. Fresh arterial or venous human blood will give no crystals. Furthermore, pigs’ and dogs’ blood dried for periods of up to eight months will give crystals which, of course, after the statement made, could not be obtained from dried human blood after any lapse of time, though the remarkable fact is given that partially dried human blood will give needles. The conclusions drawn are that crystals obtained from older stains show that it is not human blood, as does also the find- ing of tetra-hedra or prisms in fresh fluids; though the finding of needles does not bindingly indicate that the blood is human. Such a test under circumstances may be useful, but it will require further observations to make the conclusions as given vonvincing, for the question of the crystallography of the hemoglobin is one on which there is yet no accord, some writers holding that different systems of crystallization do occur in different bloods, others maintaining that there is only a variation of form in one system—that all shapes are sphe- noids belonging to the rhombic system. Certainly the same blood can by different methods be made to yield crystals of different shapes; the squirrel’s blood giving, according to methods used, either hexagons, or prisms, or tetra-hedra. Proof therefore is still to be given that this particular method will always under all variations give the same shape of crystal for the same blood. Another proposition for the recognition of blood has been brought forward by Magnamini, who makes use of the statement that oxyhemoglobin from different bloods is decomposed at a varying rate by the action of acids or alkalies, a time which may be readily determined by noting the disappearance of the absorption bands from the spectrum given by such solutions. He finds, working with certain con- centrations of solution, that the bands will disappear from human blood in thirty-eight minutes, from dogs’ blood in 110 minutes, and in other bloods after three hours or more. The results were the same with stains up to sixty days old, but after that age oxyhemoglobin became progressively less re- sistant. The poetic statement that drops of different bloods in drying on a glass plate would give different figures, each one characteristic for a certain blood, thus leading to its identification, needs only to be mentioned to show that science is not always divorced from fancy. The second part of our question, the mere detection of THE PRESIDENT’S ADDRESS (0 blood, will have to do exclusively with the hemoglobin, though it follows of course that the positive find of red corpus- cle would at once include and settle this. The detection of this body or its derivatives depends largely on its or their spectroscopic behavior, though some other tests may be men- tioned. 1. The Guaiac test, characterized by the fine blue color which a blood solution will assume, if it is treated first with a fresh alcoholic solution of gum guaiac and then with H,O, or, better, with old oil of turpentine. There can be no ques- tion about the delicacy of this reaction if properly carried out though there has been much controversy over the reliability as a test for blood. Wormley has obtained this reaction in solutions containing 1 part of blood in 50,000 and with suffi- cient fluid it will show with one part blood in 100,000. It is - Stated that stains twelve years old gave the test, though Bab- cock had unsatisfactory results with stains over three years old. Unfortunately for the convincibility of this test, a num- ber of substances give the same reaction. Among these, as having particular bearing here, are pus, bile, nasal mucus, sweat stains, and, as was found during this work, formalde- hyde, now so largely used in the arts and laboratories. The value of this test is very slight, and its failure even does not conclusively show absence of blood, since alkalies and possi- bly other reagents interfere with the reaction. Ganntner treats suspected stains with a drop of weakly alkaline water, then with a drop of H,O, solution. If blood is present there will be an evolution of gas bubbles settling into a white persistent foam. Failure indicates absence of blood but—again a restriction—pus among other substances will give the same result. The hemin or Teichmann’s test comes down to us hallowed by the lapse of time. In this test are obtained the microscopic hemin or rather haematin hydrochloride crystals, distinguished by their black or deep brown color and their form, triclinic plates, prisms frequently crossed or in clusters. The procedure is a simple one. A fragment is placed on a slide with a minute crystal of salt, covered with glacial acetic acid and heated, the crystals ap- pearing on subsequent cooling. With fluids—Struve’s method —treating first with an alkali, then tannin, precipitating with acetic acid, then treating this dried precipitate as above, seems after many trials with other methods to give the best 8 A. M. BLEILE results. The delicacy of this test must be conceded—W ormley figures crystals obtained from 1-500 grain blood and says it is possible to get them from 1-1000 grain or a fluid of 1 blood in 50,000, yet—again restrictions—iron interferes with the test so that blood spots on rusted steel could not be detected, and often too it will fail with very old stains. Further, the con- ditions essential to its success, though not in all cases fully understood, must be so closely adhered to that in experienced hands even the test may fail from undetermined causes. To quote from Babcock: “In brief, crystals of hemin, if found, furnish conclusive evidence of the presence of blood; failure to obtain them is not conclusive as to its absence.” Various substitutes—and this is indicative of uncertainty in any pro- cedure—have been proposed, as the substitution of Nal or NaBr for NaCl and formic for acetic acid, but personal expe- rience has not established their superiority over the older re- agents. Coming next to the spectroscopic tests for blood or its coloring matter, it may be said that the apparatus necessary for the prosecution of this work need be neither complex nor costly. A large spectroscope provided with a scale may be convenient and even essential for the determination of the exact location of the absorption bands, but the ones involved in this kind of research are characterized in other ways and behaviors, so that a spectrometer may safely be dispensed with. Virtually a large spectrum, that is, one resulting from great dispersion, will in dilute solution show less, on account of the spreading or thinning out of the bands, than a short one where the lines are crowded together and in which conse- quently the bands would show narrower but more intense and better defined. And, while a spectroscopic eye-piece in the miscroscope is a great convenience, practically everything can be accomplished with the small, direct vision, so-called pocket spectroscope. This may be inserted in the miscroscope in- stead of the ordinary eye-piece, and with a % or 4 inch ob- jective will give excellent results. Hemoglobin, dark red in solution, is, as already stated, the mother substance from which the other bodies here concerned are derived. It is recognized by a broad, rather dim band beginning near the yellow or I) line and extending upward to near the E line, mean 2 550. On exposure to the air the solution assumes a brighter red color, due to the formation of oxyhemoglobin, THE PRESIDENT’S ADDRESS 9 which now gives two bands, one just above D 2 579, the other just below E 1 553.8, the lower ones alone persisting in very dilute solutions. On adding a weak reducing agent the two bands disappear to make way for the one band of the again formed hemoglobin; or since the hemoglobin band is not so perceptible in extremely dilute solution it may be that the oxyhemoglobin bands only may be made to disappear and re- appear. At any rate there is here a very definite deportment, diagnostic of the presence of blood pigment, and not to be confounded with other coloring matters grossly resembling it. The test is certain and quite delicate. The intensity of the bands will of course depend, (1) upon the strength of the solu- tion; (2) upon the thickness of the latter, or, what amounts to the same thing, the width of the slit in the spectroscope. With the appliances mentioned blood may be detected in a layer 15 mm. deep diluted 1:4000 or in a 40 mm. layer 1:5000, the actual quantity of fluid used being in the latter case equiva- lent to .0003 c.c. of blood, in the former .0001 c.c. However, as said repeatedly, hemoglobin is a fugaceous substance, and in fluids not neutral, or in old stains or heated or washed ones, this body has been decomposed, leaving commonly the hematin previously mentioned. This substance, soluble in acids and alkalies by means of whose action on hemoglobin it is obtained in the laboratory, also has a definite spectrum, its lower absorption band lying close to the D line. Spectro- scopically hematin is less sensitive than oxyhemoglobin, not showing in dilutions greater than 1:1000 (15 mm. layer), yet it possesses properties which make it admirably adapted as a witness in this question, and on which properties is based the propesition of the method for blood examination to be sug- gested. Unlike hemoglobin, hematin is a stable body not readily affected by agencies to which blood-containing fluids or spots are usually subjected. From it Hoppe, Seyler and Stokes first obtained by reduction a body known as reduced hematin—or better, hemochromogen—with its own spectrum and other properties which make its identification certain be- yond any doubt. For the production of hemochromogen the following method is well fitted: The solution or substance is treated with KHO solution 5¢, using heat with old and dried material. To the solution is added pyridin (1-10 its volume) and (NH,).S, when the previously greenish solution will turn cherry red and remain so if kept from the air. In the spec- 10 A. M. BLEILE trum can now be seen two well-marked extraordinarily intense absorption bands, the lower one of which is the more persist- ent and which alone need be considered here. It lies midway between the D and E lines, mean 2 557. On shaking with air the bands disappear to come again on resting the fluid. In this way can blood be detected, with a 15 mm. layer, in dilu- tion of 1 blood to about 20,000 of water, or, with a 40 mm. layer, 1 in 40,000 involving of actual blood about 5-100,000 c.c. The certainty and characteristic of this test is equal to that of oxyhemoglobin and it is from three to four times more delicate. Inquiry as to the availability of this test where the material to be examined had been so treated as to lead to a greater or lesser decomposition of the blood, was suggested by a case necessitating such an investigation under peculiar circum- stances. A triple murder had been committed, the instrument used being an ax shown to be the property of defendant in the trial. After the murder the house, a wooden one, contain- ing the bodies, was set on fire, burning to the ground. The ax had been thrown down outside about eight feet from the house, thus being subjected to a high heat, as further shown by the charred remains of the ax handle, which was burnt up into the ax. On the ax were found charred and brittle hairs and some brownish black spots which, if blood, were too much altered to yield hemoglobin and on account of the iron rust would presumably fail to give the hemin crystals. To test the resistibility of the coloring matters, though not bear- ing directly on this case, blood was first treated with various chemicals known to affect the hemoglobin, such as 10% solu- tions of KHO, NH,O, HCl, HNO,, H.SO,, HgCl., strong alcohol] and formalin. After six months’ maceration in these fluids it was easy to obtain, by the method given, the hemo- chromogen reaction with all its essential points. To test the influence of heat, dried blood was heated for ten minutes at temperatures from 100° up to 280° centigrade, and in all cases the reaction was obtained. though the color of the pyrogenous bodies formed at the highest temperatures em- ployed interfered with the spectroscopic examination and forced great dilution of the fluid. Age of material does not apparently interfere with this test. At least the bands were obtained from a stain on cloth sixteen years old and not over one mm. in diameter. Having in mind then the ease of appli- THE PRESIDENT’S ADDRESS 11 cation, delicacy, certainty, and freedom from influences by many disturbing agencies, of this test, an outline of a con- venient method for the detection of blood would be as follows: After an attempt to find red corpuscles, and with or against success in this direction, without wasting further time or material which may be at disposal in small quantities only in a search after the less delicate or less persistent hemoglobin or the hemin crystals so liable to fail, the substance is to be at once treated with KHO solution, heating if difficult of ‘solution or not already dissolved, and then adding pyridin and (NH,).S, as previously outlined, and observing the spec- trum. Where a small stain on a thin fabric is the object of study it can be placed on a cover glass, moistened with a drop of KHO solution and pyridin. After some minutes a drop of (NH,).S is to be added, the preparation inverted over a hollow ground slide, sealed with oil, and placed under the microscope, when the spectrum will show the hemochromogen band, disappearing on exposing the preparation to the air, if blood is present. t v4 ay ah tye lt, + re J AAA SOME ADVANTAGES OF FIELD-WORK ON SURFACE WATER SUPPLIES By HORATIO N. PARKER, BIOLOGICAL LABORATORY OF THE METROPOLITAN WATER WORKS OF MASSACHUSETTS There are several ways of procuring a water-supply. The simplest is by catching the rain as it runs from the roofs of houses and storing it in cisterns. Where provision has been made for passing the first portion of the rain, saving only that which falls later, and when the cistern has been carefully built, this is a satisfactory method. But the amount that can be obtained in this manner is small, and serves, at the most, to supply only two or three houses. Another source of supply is the underground water which is derived from springs, wells, and filter-galleries. Carefully operated works of this class usually afford a water of excellent quality and one which is very satisfactory to the consumers. Unfortunately, however, it, too, is limited in quantity, and besides, as larger demands are made on it, tends to deteriorate, becoming constantly harder and at the last carrying bacteria in high numbers. By far the greater part of our cities are supplied with surface water which is taken from rivers,—with or without filtration,—from natural lakes, or from artificially constructed storage reservoirs. It is the purpose of this paper to point out the necessity of carefully conducted field-work in those places whose supplies are obtained from the two last sources. As the rain falls to earth, it washes the dust, the spores of microscopic organisms and the bacteria from the air, and absorbs from it various gases. But these impurities are trifling as compared to those it acquires after it reaches the ground. The character of a water is determined by the water- shed from which it is collected. To make what follows perfectly clear, let us imagine a watershed which comprises all the features commonly found in the localities on which water-supplies are built. It has an area of about one hundred square miles and a somewhat 14 HORATIO N. PARKER diversified topography. There are a few high hills, one of which is crowned by a large town, which is sewered, and whose sewage disposal works are outside the watershed. Through the town runs a brook which flows onto English filters at the shore of a lake which forms part of the system of water supply. The effluent from these filters runs directly . into the lake. Ali of the hills slope abruptly to the flat land at their base. A river with its numerous tributaries courses through the middle of the watershed. On one of these streams whose drainage area is wild, wooded, gravelly land, and which has its rise in dense cedar swamps, a storage res- ervoir is built. A second and larger tributary flows through a cultivated district whose soil is a loam mixed with con- siderable clay. Here and there are farm houses with the ac- companying live stock, and some distance away is a hospital with its own water supply and English filters for sewage disposal. This stream has been dammed at three points, and each of the storage reservoirs thus formed has its own tribu- taries which for the most part are fed by springs. All of the brooks enter the reservoirs at the head, and on some of them dams have been built to form mill-ponds. These are now abandoned. The soil has been stripped from the bottoms of all the storage reservoirs, which vary from fifteen to fifty feet in depth, and have a capacity of two billion gallons. They are constructed so that water can be drawn from the surface, mid-depth and bottom. The lake is somewhat isolated from the storage basins; it has a capacity of three billion gallons, and the bottom has never been cleaned. The mains are built so that water can be drawn from all the supplies at once, or any source may be used independently of the others. No houses are allowed on the shores of these ponds, and the whole dis- trict is under rigid sanitary supervision. Let us first consider the chain of reservoirs which are built on the stream flowing through the inhabited area. The first rain that falls after a long dry spell will be soaked up by the earth, but as the storm continues the capacity of the ground to do this becomes exhausted, and the excess of water must flow away over the surface. Not in clear rivulets, how- ever, but in very dirty ones, for the continual beating and pelting of the rain loosens the clayey soil so that it is easily dislodged and carried away by the water toward the reser- voirs. Moreover, we have said that the valley of the stream , FIELD-WORK ON SURFACE WATER-SUPPLIES 15 is cultivated; this necessitates manuring, and if there are market gardens, heavy manuring, which means that some and possibly much night-soil is used. Thus a chance is afforded for disease germs to be washed into the reservoirs, and in any case many bacteria and nitrogen in the form of nitrates will be gathered from this source. So these little rivulets flow on with their ever increasing burden of mud and putrescible matter and deposit it in the feeders of the reservoirs. Of course the gross polluting centers, such as barnyards, privies, etc., have been kept from draining directly into the supply. It is only the small waste incident to life, to which man and beast alike contribute, that is carried into the storage basins. The swollen feeders are now adding bacteria to the reservoir and mud to make them turbid. This is an objectionable state of affairs, for from the head of the reservoirs the roily water, high in bacteria, moves onward till it is delivered to the consumers or passes out of circulation. Its progress through the reservoirs must be care- fully watched. It is not enough to wait till samples collected at the intake announce its appearance there. The superin- tendent not only ought to know the character of a water he is using at a particular time, but he should be kept informed as to whether ?#t is likely to improve or deteriorate in quality. Turbidity is a deterioration, and is justly complained of by the consumers; not that it is injurious to the health, but for aesthetic reasons. It makes the water unsightly, and imparts an earthy flavor to it. The degree of turbidity de- pends upon the amount of clay in suspension. The sand parti- cles soon settle out, but the clay, which is very finely com- minuted, does so very, very slowly. If a little clay is brought to the supplies by a short rain, it may sink to the bottom be- fore it reaches the mains, unless the reservoirs are kept stirred up by the wind. On the other hand, a severe storm will bring much clay, which will gradually pass through the reservoirs to the intake. It is evident, that careful records of the clearness of the water should be kept. There are several ways of estimating turbidity. The oldest is by determining the weight of sus- pended matter in a given weight of water. This method is being abandoned because of its inaccuracy. A small amount of sand would raise the weight of suspended matter very con- siderably without a corresponding gain of turbidity, while 16 HORATIO N. PARKER a large addition of clay would cause a marked increase of turbidity without materially increasing the weight of matter in suspension. Nowadays, the commonest ways of measuring turbidity are by means of the Silica Standards,* Diaphanometer,; and the Platinum Wire.t Each has its advantages, and that must be selected for use which is best adapted for the work in hand. Still another method was formerly employed on the Metropolitan Water Works; a disc,§ five inches in diameter and painted like a surveyor’s target, is lowered into the water, and the greatest depth at which the divisions can be distin- guished is recorded. In reservoirs several factors work toward reducing the bacteria. They tend to sink to the bottom, and, besides, the water is not as favorable a medium for them to grow in as is the land from which they have been washed; further, the sunlight-is inimical to them. So altogether they fare badly after they reach the supplies, and the chance of their reach- ing the intake are somewhat less than that of the clay. It is well after a severe storm to make a thorough examina- tion of the supplies. So we go in a boat from the foot to the head of the reservoirs, taking turbidity readings and bac- teria samples at frequent intervals. The bacterial examina- tions are mainly quantitative, supplemented by tests for coli reactions. It must be so, as a search for disease germs among the host of other bacteria would be as futile as the search for the needle in the haymow. This work serves to let us know the condition of the reservoirs soon after the rain. A few days later the trip should be repeated and, by comparing the results obtained then with the former ones, we can tell whether the chances favor increased bacteria and turbidity at the intake or whether they will disappear without occasion- ing disturbance. If one of the reservoirs is in an unsettled condition from these causes we shut it off for the time being, and draw from one which appears to be normal. In this way complaints from consumers can be avoided, and a feeling of security established in the community which would have been impossible without the field-work. We have taken up the two main factors which menace the * Technology Quarterly, Vol. XII, No. 4, p. 283. + Technology Quarterly, Vol. XII, No. 2, p. 145. t{ Hazen, Filtration of Public Water Supplies, 3d edition, p. 118. Whipple, The Microscopy of Drinking Water, p. 75. FIELD-WORK ON SURFACE WATER-SUPPLIES 17 supply in the inhabited district, but there remains to be con- sidered the abandoned mill-ponds and the hospital. The mill- ponds will be spoken of later. Let us turn our attention to the hospital. It is situated on high ground at some distance from the chain of reservoirs, but the effluent from its disposal works must finally find its way into the supply. For this reason it is imperative that the English filters at all times do their work perfectly. It will not do to trust entirely to the hospital authorities in this matter. They will undoubtedly be honest in their intention to run the filter beds properly, but the best of employes grow lax at times, and accidents will happen so that he who is responsible for the purity of the water supply should be able to say that at such and such a time the beds were doing thus, and so. Much tact is neces- sary, in a case of this kind, where a third person must concern himself with another’s business, but if all approach the matter in the right spirit there need be no friction. The field-work on the storage reservoir in the wild land takes on a different nature. As the soil is gravelly there will be little additional turbidity after rains, and as it is unin- habited no great increase of bacteria that may be objection- able. But this reservoir, in a small way, but in a serious one, is liable to contamination. We have indicated that it is wooded; its chief feeder rises in a cedar swamp, and its shores are covered with deciduous trees. This being so, it will be a resort for picnic parties, and for any one who wishes a day’s outing. They should not cause trouble, but unclean peo- ple will be among them, and they will not fail to arrange mat- ters so that some foecal matter will find its way into the sup- ply. The sick, and not the well, will offend oftenest. The danger is a real one, and the only relief is to have the shores so thoroughly policed that sinning will be difficult. It will not do to rely for protection on bacterial examinations. As has been said before, it is next to impossible to distinguish the disease germs among the shoal of other bacteria. We must keep them out of the supply, and not rely on hunting them down after they are once in. It may not be amiss to speak here of the danger of allow- ing skating, fishing, and camping on bodies of water which are used for drinking by man. The evil is the same as in the case of the picnickers, and so these sports should be pro- hibited. A public water-supply is not a plaything, nor a play- 9 —_ 18 HORATIO N. PARKER ground; it is an extremely sensitive and expensive plant, built to administer to certain definite needs of the entire people, and should not be sacrificed to the pleasures of a few. Of the same kind of nuisance is the inexcusable and unnecessary custom of manuring the sides of reservoirs to get thick grassy | slopes. The practice is a filthy one and dangerous besides, for in the majority of cases the origin of the manure is un- known, and it is not at all impossible that it contains disease germs. If it does the wind and rain will surely not leave them on the shores. When enrichment is desired it would seem per- fectly feasible to use some of the cheap chemical fertilizers. But to return to the reservoir once more. Color is the other point to be considered in regard to this source. We have said that the feeders rise in swamps; in flowing through them they will acquire a deep red color. This will be more noticeable some times than at others. When the water flows rapidly through the swamps, it will be much lighter colored than when the flow is sluggish, and time enough has elapsed for it to leach out the coloring matter from the roots and peat. In the late spring it will be particularly dark, for the water which comes out then has remained backed up in the swamps all winter, and besides the leaves which were shed by other trees than the evergreens the autumn before have disinte- grated and their coloring matter is added to that from other sources. _ When the influents reach the head of the reservoir, their course depends on their temperature and that of the reservoir itself. If the temperature of the feeders is such as to make their density less than that of the reservoir, the dark influent, unless disturbed by winds, will flow over the surface of the reservoir to the intake, arriving there almost as dark as when it entered at the head. If the density of the influent is greater than the reservoir water, it will flow along the bottom; if they are of the same density or high winds prevail, their waters will commingle. Sunlight bleaches the water somewhat so that the reservoir is likely to be lightest colored at its foot. At best, the water is too dark to be used by itself without dis- quietude on the part of the consumers; so when distributed it must be mixed, in such quantities as not to cause comment, with the light colored water of the supply. This is very nice work, and it makes it necessary for the color of the water to be accurately determined. FIELD-WORK ON SURFACE WATER-SUPPLIES 19 Not only must color samples be taken, but careful record of the temperature of the reservoir throughout its greatest depth must be kept, together with that of the influent streams. At the outlet color readings must be made on samples from the surface, mid-depth and bottom, in order that water may be drawn at that point where the color is the lightest. Also the color of the infiuents must be determined besides that of samples taken at various points intermediate between the foot and head of the reservoir. This is done to trace the progress of the water through the basin. All this work occa- sions frequent visits to the supply, but the time is profitably spent, for no change in a water is so quickly noted by the citizens as that in color, and superintendents value highly the information which enables them to anticipate criticism from this cause. Turning our attention next to the lake, we find as its most striking feature the proximity of the town on the high hill. The town is a constant threat to the purity of the lake. To be sure, the sewage is carried outside of the watershed, and the brook, which takes almost all of the surface drainage which escapes the sewers, is filtered. But the town with its busy life is there, and if an accident should happen to the sewerage which should let the sewage unobserved into the lake or some similar misfortune should occur, trouble would surely ensue. The best the water works managers can do is to watch the filters carefully, and to be constantly on the alert for escaping sewage from the sewerage system. Those places where it passes near brooks which empty into the lake should be especially guarded. Bacteria samples should often be taken from the brooks, and frequent trips for inspection should be made along their banks. To watch the filters is a comparatively easy matter. In the first place the plant should be in the hands of a competent person. Then bacteria samples should be taken from the effluents at regular intervals, and at any time the operator of the station may see fit to suggest. Whenever bacteria sam- ples are taken from the effluents a bacteria sample and a sample for microscopical examination should be taken from the applied water. As the lake is fed by springs, and as the only important influent is filtered there will be little trouble from turbidity and color. We have now taken up the salient features of each source 20 HORATIO N. PARKER of supply which necessitates peculiar field-work on their watersheds. One characteristic which is common to all re- mains to be discussed. I refer to the growth of microscopic organisms and the tastes and odors caused thereby. When we first began to strip our storage reservoirs, we found that we had removed the organic matter so thoroughly that large growths did not exist, and we hoped this condition would be permanent. Experience has proved otherwise. The organ- isms have gradually established themselves in reservoirs which were built in the most painstaking manner. Many causes contribute to this result. Perhaps the three most prominent ones are the increase of population on the water- shed, the slow accumulation in the reservoirs or organic mat- ter from the feeders, and the bringing in of microscopic forms by these same brooks. So we must acknowledge the growths to be common to all reservoirs. Not in equal degree, however, for we ordinarily expect them to be smaller, and to occur less frequently in the basins where the cleansing has been most perfect. It is the duty of the water works biologist to know the organisms that cause trouble, to study the conditions under which they occur, and to so draw from the supplies at his command that the water served to the community shall be palatable and wholesome. He can never do these things satisfactorily unless he has an intimate personal knowledge of the system, and this can only be gained by much field-work. It will not do to rely entirely on the analyses made in the laboratory of samples collected by another. They give no idea of the distribution of a growth, and at times are very mislead- ing as to its size. The collector may be attracted by little specks in the water and, knowing that they are of interest to the analyst, try to get as many as possible into the bottle. Or he may have been warned against doing this, and so, almost unconsciously, try to keep the sample clear. Again, he may be very much hurried, and take the sample without noticing the condition of the water at all, it being merely a matter of luck whether it is a representative one or not. In- deed, analyses of samples taken in this manner simply show what the collector dipped up. < —_____—_jg98 1896 Pen Ce e « D . ” yi ¥ 6 2 a i ¥ « ay a c- ® & - > oh 7 = — \ = ’ * é 7 . x a — C z - 7 s e am i “ METHODS OF PRODUCING ENLARGEMENTS AND LAN- TERN SLIDES OF MICROSCOPIC OBJECTS FOR CLASS DEMONSTRATIONS By JOHN ASPINWALL For convenience we will divide the subject of this paper into three parts: 1. Making the Photomicrograph. 2. Making the Lantern Slide. 3. Making the Enlargement. MAKING THD PHOTOMICROGRAPH The method used by the writer is the result of an attempt to produce photomicrographs of considerable magnification, and yet of great depth of focus, while using lenses of high re- solving power. In the ordinary process of photomicrography, the amplifica- tion is obtained in one of three ways: 1. A low power objective, and fairly high power ocular. 2. A high power objective and low power ocular, or none. 3. A great extension of the bellows of the camera, combined with the use of a projection eyepiece, or none. In all of these cases, amplification is obtained at the ex- pense of the focal depth; and, although there is often defini- tion over an extended area at right angles to the axis of the beam of light, the relation of the parts is only shown over a very thin area in the line of the axis of the instrument. The method of the writer is to use an objective of medium power, where fairly high amplification is desired, say a one- quarter Spencer, making the negative of a diameter of 13 to 2 inches. In a lantern slide camera, enlarge from one-half to three- quarters of the central area of its negative to twice its diameter upon a Paget lantern slide plate. By another en- largement from this positive, a negative of any diameter can be secured. 42 JOHN ASPINWALL This appears a roundabout method, but the object is ob- tained, viz., to get the maximum depth of focus and any de- sired amplification. It is important to make the positive upon a plate rich in chloride, such as the Paget, in order to obtain a deposit with- out grain, and capable of registering the minutest detail with no suggestion of film structure. It is also important that the enlarging lens be of the very best type, such as a Ross, a Guerz, or a Zeiss, and well stopped down. in making a negative for an enlargement on bromide paper, the same methods may be pursued. The writer prefers, in making the original negative, to use an objective without the ocular, and instead of the usual substage condenser of high angle, to employ an ordinary ob- jective of, say, one inch focus. For the best results, the beam of light approaching this objective condenser should be of a very low angle. This can be obtained in the ordinary photomicrographic outfit by the interposition between the lantern condenser and substage of a biconcave lens of suitable curvature. MAKING THE LANTERN SLIDE First of all, no American plate known to the writer is capable of producing the best grade of lantern slide. We pre- fer the Paget plate, which is made in England. It is important to depart from the beaten path, and leave, in most cases, the black and white effect, and reach into the warmer tones of brown, purple and red. This may sound esthetic, but the fact remains that the subject is given a look of life and substance by adopting a warmer tone. Nothing but a suggestion of death lurks in the chalky white and black tone of the ordinary commercial lantern slide. It is certain that two slides of a hand suffering from skin disease—one in black and white, and the other in a tone near to flesh tint—bear no comparison; one is the hand of the dead, the other that of some living being. In the matting, too, there is much to do with the ultimate results. This point cannot be too strongly enforced. Any subject surrounded by a mass of glaring white light will fail to show the most delicate lines and gradations of tone, owing to the eye being dazzled by the surrounding whiteness. ENLARGEMENTS AND SLIDES OF MICROSCOPIC OBJECTS 43 The proper mat is one capable of being cut to suit the sub- ject; such an one, for instance, as that known in the photo- graphic world as the Boston mat. The lines given on this mat enable one to shape the opening to suit the subject. The use of the mat of ordinary size, such as usually employed by the commercial lantern slide maker, would seem to indicate that the value of the slide was in proportion to the area of the opening; while the reverse is really the case in most in- stances. The Paget slow plate is capable of giving tones from black, through the browns and reds, into the purple. The developer used by the writer is made up as follows: Fiydrochinon yen. ine ecleriecee 100 grains Sodium sulphite (crystals).......400 grains Sodium carbonate (crystals)..... 400 grains WWiaiers Mater a aiunent ate a lareenty wal eearatetay « 20 ounces The exposure will run from 20 seconds to 15 minutes, ac- cording to the light or the tone desired. A long exposure and a weak developer, with bromide added, producing the warmer tones, and a short exposure, with strong developer, the blacks and whites. The dilution of developer is made as follows: 1. For black tone— BEVEL ER oa a et ek sigh iohette tale Lets 1 ounce WALGER ES Mos tunes sien pata mia ais to i cps avers 2 ounces 10% solution of bromide........ 1 to 2 drops 2. For brown tone— We wela Meng. ss ee o eae Book, gal 1 ounce IVF ATS Pi heoigs: fa Gist taketh Miche ARIAS oo Lin's wile 4 ounces EONAR es ph esialesies o4) dz lai OX LOS OLGDS 3. For red tone— UD Te PEI Ci eS ei Ae En 1 ounce WicGletiaameieer roy serch. ek ls Uk tha 8 ounces PREOUI GS ie ee IL es oll ows! 15 to 25 drops 4. For purple tone— Over exposure and the use of the No. 2 dilution. As Hydrochinon is inert at low temperatures, for uniform results the developer should be slightly warm in winter, so that it will be between 70 and 80 degrees F. 44 JOHN ASPINWALL The exposure, is made, say 20 seconds for black tones, 14 to 2 minutes for brown tones, and from 5 to 15 minutes for red ones. The quality of the negative counts, of course, and no rule can be given—the foregoing exposures being merely suggestive. The reader will have to work out the problem from the hints herein given. The time of development increases as the tones get warmer. The writer has obtained the best results in making slides by using a reducing and enlarging camera in preference to making slides by contact with the negative. It must be apparent to anyone that a slide from a photo- micrographic negative 24 inches in diameter will be superior in depth of focus and detail to one made by reducing a nega- tive six inches in diameter made of the same subject and area of subject where this increased size of negative is obtained by drawing out the bellows, or using a higher power objective. Quite a remarkable effect can be obtained in some cases where tissues are differentiated by highly stained nuclei, in the fol- lowing manner: A lantern slide is made with the reddest tone obtainable. After fixing and washing, but before the slide is dry, it is toned for a short time in a gold bath made as follows: 1. Sulphocyanide of ammonium... .200 grains The Ss eh OETA eee ee RAN 32 ounces Carbonate of soda (granuls)..... 2 grains 2. Chloride of gold (brown)........ 15 grains WWW OR eve tesnes et eis siete Woke! Lot 1 ounce For use take two ounces of No. 1 and four drops of No. 2, always remembering to add No. 2 to No. 1, and never revers- ing the operation. This amount of solution will tone one slide to a perfect blue throughout; but in our process, we only immerse the red slide, before spoken of, long enough to permit the gold to attack the lighter deposit of silver in the film. The result of this will be to give the lantern slide an appearance of a microscopic slide, which has a nuclear stain of carmine, and the deepest stain of methyl blue. The gold bath should be kept at a temperature of from 72 to 76 degrees—a lower temperature would result in failure. It would seem that the effect we obtain is not altogether permanent, as, after a couple of years or so, the blue appears ENLARGEMENTS AND SLIDES OF MICROSCOPIC OBJECTS 45 to gradually creep into the red nuclei and spoil the differ- ential effect. You must not fail to remember that in all these colored effects, any shadow in the negative caused by refraction will assume solid proportions when interpreted in the color of the subject. This, however, holds true somewhat of a black de- posit. I would advise using the more brilliant tones only upon subjects made with a low power objective, unless it be of a very thin section, and an image which it is clear cut and free from exterior refraction lines. A finely hand colored slide is probably the most perfect for the purpose of class demonstration, but such slides are very expensive, and require some time to prepare; while chemically colored slides are quickly produced, and cost practically no more than the ordinary kind. There is one point which is important in producing the very best grade of lantern slide with the Paget plate. This is the clearing operation which must follow the fixing. The devel- opment should be carried a trifle further than normal, and after the slide is fixed, but before washing, swab it over with a tuft of cotton immersed in a weak solution of ferricyanide of potassium (red prussiate of potash) of the color of very pale sherry. This will remove all chemical fog and clear up the whole image while reducing the entire deposit a trifle. Areas too prominent and calculated to divert attention from the greatest points of interest, may be either reduced or entirely wiped out. For instance, in the cross section of the skin, and the tissues below, it frequently happens that the portion outside of the tissue proper shows refraction areas, or specks of dirt, and bits of the tissue. These can be entirely removed by rubbing gently with the cotton dipped in a strong solution of ferricyanide, being careful to hold the slide so that the solution will not run down onto the image of the section. {t is frequently the case in sections through the skin and below, that the epidermis is brought out with too much prominence; while the tissues below are the subject of dis- cussion, and therefore, it is wise to reduce this superficial layer to a density which will not attract the eye away from the main subject. MAKING THE ENLARGEMENT A convenient form of enlargement for demonstration in a 46 JOHN ASPINWALL small class is a circle of about 18 inches in diameter, mounted upon a very heavy square card with a white margin of about an inch on each side. These can be either set up before the class, or handed around. The method of producing these does not differ materially from that usually employed. I use a rather weak negative, such an one as would give an Aristo print of fine gradations, with no portion very dense. Parallel light is obtained by means of an are lamp and a condenser so arranged that the arc is at its focus. Only the central portion of the condenser is used; i. e., we would use an 8-inch condenser to project a negative image of not more than 4 inches in diameter. The finest medium focus, double series, view lens, is used for projection, and it is well stopped down, say to F.-16. A paper made by Eastman, of quick speed for the class of negative employed, is tacked to a board absolutely at right angles to the axis of the beam of pro- jection, and enough time is given to insure the obtaining of every detail of the image. Where there has been oyer-staining in certain areas of the sections, portions may be shaded to prevent false effects. Developing is done in adurol: one portion of developer to about 30 of water, and bromide added according to the character of the image required. After development, wash and place in a weak solution of hypo, with a saturated solu- tion of chrome alum added, say in the proportion of 1 to 20, in order to prevent blistering. The quantity of chrome alum to be added depends some- what upon the temperature. A solution of formaldehyde, made very weak, may be used after the print has been par- tially washed upon removal of the hypo solution. Wash thoroughly and hang up to dry. Adurol, if properly handled, gives a brownish tone just off a black, and adds life to the enlarged image. In making the enlargements from a negative with clear glass surrounding the round microscopic image, a piece of dense paper, preferably the yellow post office paper, is cut to whatever size we desire, and placed back of the negative so as to cut off the light of the arc lamp from the surrounding area. It is well to reduce somewhat the size of the image upon the negative by allowing the paper to lap down upon the image. This gives a clear cut edge to the circle when enlarged ENLARGEMENTS AND SLIDES OF MICROSCOPIC OBJECTS 47 upon the bromide paper. The writer believes that if this system is followed out, with such modification as may occur to the manipulator, the result for class demonstrations with lantern slides and enlargements will be superior to that now generally obtained. ON THE DISTRIBUTION OF GROWTHS IN SURFACE WATER-SUPPLIES AND ON THE METHOD OF COLLECTING SAMPLES FOR EXAMINATION By FREDERICK S. HOLLIS WITH FOUR PLATES The purpose of the study of the micro-organisms floating in a body of water may be two-fold. It may be conducted for purely scientific information or for practical purposes, as a means of determining the total amount of material which is available as food for higher forms of life, or the results of the study may be used as a guide in properly conducting a system of water works. In the case of the study of the micro- organisms in connection with water-supplies, they are to be regarded as deleterious, and the determination of the exact position and recurrence of growths becomes of the utmost importance as a means of avoiding them. For such practical ends in this connection, the determina- tion of the micro-organisms is only a part of the necessary study, and such determinations should be supplemented by chemical and bacteriological examinations of the water. The micro-organisms are, indeed, to be considered only as a phase or form of the organic contents of the water, which, in this form, is objectionable as a source of odor and taste caused either by the characteristic odor of growth of the particular form or resulting from its decay, and as a source of food which will during its decay give rise to an abnormally large bacterial growth. The relation between the micro-organisms and the other forms of impurities of a water is best seen in a study of the nitrogen contents of the water. Starting, say with an organic growth, the nitrogen is in combination with the other constituents of the organic bodies and appears in the chemical analysis as albuminoid ammonia. After the death of the organism it becomes disengaged as a result of decomposition and exists, first as free ammonia and, as the 4 50 FREDERICK S. HOLLIS result of various states of oxidation, as nitrites and nitrates, in which last stage it is available as plant food to be built up again into organic bodies. Individual samples must be taken for the chemical and bacteriological examinations and, in order that the com- parison may be made between the organic life and the im- purities in the other forms, the samples for microscopical examination must be identical with those taken for the other examinations. Microscopical examinations have been made for the past ten years of the water of the reservoirs of the Massachusetts Metropolitan Water Works, which passed from the control of the City of Boston on January 1st, 1898. Samples are taken regularly once a week from the surface, mid-depth and bottom at the deepest point of the reservoirs, which is com- monly near the gate-house or outlet. These results have been supplemented, when necessary, by samples taken every few feet and, during periods of growth, by regular inspection of the sources and the collection of samples at various parts of the reservoir, as a means of determining the rate of exten- sion of growths through the reservoir. Chemical samples are taken less frequently and also ocea- sional bacterial samples at the surface, mid-depth and bottom for comparison. The results obtained from the samples collected in this way and their usefulness as a means of avoiding growths which would be objectionable if taken into the distributing reservoirs have convinced us that the information which is most to be desired is best obtained from such samples. The samples for the microscopical and chemical examina- tion are taken by lowering a collecting bottle to the desired depth in a weighted cage and, by means of a separate cord, withdrawing a cork stopper which has been substituted for the ground glass stopper. The neat form of collecting cage in which a spring releases the stopper, thus making unneces- Sary a separate cord, was devised by Mr. G. C. Whipple. The eight reservoirs of the Metropolitan Water Works offer uncommon advantages for the study of the surface water of that section of New England. Each receives surface water colored more or less according to the season of the year by the peaty matter of the valley through which the influent flows, but almost entirely free from the turbidity a eo. GROWTHS IN SURFACE WATER-SUPPLIES Dil caused by the presence of clay, which is noticed in other sections of the country. Such slight turbidity as is caused by the spring rains is due largely to the presence of fine sand or rock flour and subsides so quickly that it rarely reaches the outlet end of the reservoir. Lake Cochituate is formed of a chain of three lakes which were deepened considerably by building a dam across the outlet of the lowest one fifty years ago, when water was first taken from this section for the supply of the City of Boston. Whitehall Reservoir was also formed by enlarging a natural pond, but it is deepened to such an extent that it is prac- tically an impounding reservoir. Framingham Reservoirs Nos. 1 and 2 were formed by constructing dams across the main stream of the Sudbury River. Sudbury Reservoir, Framingham Reservoir No. 3 and the Ashland and Hopkinton Reservoirs were formed by building dams across the various feeders of the Sudbury River. Water from the south branch of the Nashua River, which will eventually be impounded in the largest reservoir of the series, has been collected for more than two years by means of a temporary dam and deflected to the Sudbury Reservoir, the largest present member of the series, of which it has become the principal feeder. Depth, when Contents in filled, at billion gallons. deepest point. Mae y OCH IAUWAEE) |: io5) ce sete s Mab dao aes 2.9 60 ft. Framingham Reservoir No. 1.......... 0.3 15 Tt: Framingham Reservoir No. 2.......... 0.5 Wa EG, Framingham Reservoir No. 8.......... b2Z 21 EC. SME NCHEE VOI i c.5/ 2.4 sricye veal eieis aie a, 6 7.6 about 55 ft. FPOPKIMEGN, TRESEEVOIT * 5 .)5.0)056 6 «cw ses oasis 1s about 52 ft. POM MEA FRESE MONT ne oc) 50° van, e 55.0 0 3 6 aha 1.4 49 £t. Whitehall Reservoir 2... b. cece eee 1.6 25. Lt The water from the Nashua River, together with that col- lected from the water shed of the Sudbury Reservoir passes, after storage for a considerable period, through Framing- ham Reservoir No. 3, the next lower reservoir of the series, to the entrance of the pipe line and aqueduct leading to Chestnut Hill Reservoir. Water from the other reservoirs passes through Framingham Reservoir No. 1 and to the same 52 FREDERICK S. HOLLIS aqueduct. A separate aqueduct leads from Lake Cochituate to Chestnut Hill Reservoir. From Chestnut Hill Reservoir it runs directly to Boston in pipes or is pumped to the various distributing reservoirs of the Metropolitan district. All save Lake Cochituate and Whitehall Reservoirs have gates for drawing the water from both the surface and bot- tom, and the deeper and more important ones have also a gate at the mid-depth. The surface soil has been completely removed from the entire area of the more important reservoirs, and in some cases the influent streams have been diverted from the swampy areas. which caused an increase of color, by ditching. The water of a few of the brooks, more likely to be contami- nated than the others, ‘is filtered before it is received into the reservoirs. All of the examinations haye been made by the Sedgwick- Rafter method which commends itself because of its accuracy and the comparatively small factors used in converting the recorded results of the observations into standard units per ce. The Jackson funnel is used and the degree of concentra- tion most commonly employed is 500 to 10. All results are expressed in terms of the standard unit per cc. (1 standard unit—400 sg. microns) as proposed by Mr. G. C. Whipple. Results expressed in standard units per cc. are an approxi- mation to a quantitative estimation in which the same num- ber of standard units of the different forms express as nearly as possible equal amounts. Results so expressed agree more closely with the results of chemical analysis than those ex- pressed in numbers per cc., and are to be preferred greatly for accuracy and usefulness. From a study of the growths of the principal reservoirs it is seen that they may be divided into groups which show a different development and distribution of growths. In those in which water is collected and held until used, the water is quiescent except for the action of the wind and the overturn at spring and autumn due to temperature changes. In such reservoirs the development of the growths is a normal one and, in general, a marked difference is noticed between the abundance of the organisms at different depths. In those in which the water passes through the reservoir at a considerable rate, growths are brought in and mingled with those of the reservoir and a normal development is prevented by the circulation of the water. GROWTHS IN SURFACE WATER-SUPPLIES 53 On the accompanying plates this is shown by the average of weekly analyses from 1895-9 inclusive for six of the reser- voirs. (Plates V and VI.) Calling the average number of organisms for the year of each source at the surface as 100, the following table shows the average yearly number of organisms of the mid-depth and bottom of each source expressed in percentages of the surface growth: SURFACE MID-DEPTH BoTTom Per cent. Per cent. Per cent. Organ- of Organ- of Organ- of isms surface isms surface isms surface Sudbury Res. ....339 100 226 67.5 169 49.2 Hopkinton Res. . .550 100 276 50.2 214 38.9 Ashland Res. .....178 100 123 69.1 95 53.5 Lake Cochituate.. .672 100 576 84.9 608 88.0 Fram. Res. No. 2. .158 100 143 90.5 107 67.9 Fram. Res. No. 3. .581 100 510 87.7 487 83.8 The Sudbury, Hopkinton and Ashland Reservoirs belong to the first group in which normal growths are possible and do occur. Framingham Reservoirs No. 2 and 3 are as ordinarily conducted members of the second group. ‘The organisms of Framingham Reservoir No. 3 have been much lower since water has been supplied from the Sudbury Reservoir and the Nashua River than when filled with water from its own water shed. Lake Cochituate, while it would seem to fall under the second group, does, in reality, belong as far as most of the growths are concerned to the group in which there is a normal development of growths. Several causes act to make the average number of organisms irrespective of species similar at the surface, mid-depth and bottom. The lake at its deepest point where samples are collected is sixty feet deep and the bottom at this point is ‘such that marked stagnation effects follow the quiescent state of the water during the summer and to.a lesser extent during the winter when the surface is covered with ice. When the water at the surface reaches the temperature of greatest density which commonly hap- pens in November and again in the spring soon after the ice leaves the reservoir, there is a complete mixing of the water of all depths. Crenothrix, which has become abundant at the bottom, is brought up and distributed quite evenly throughout the water at all depths. 54 FREDERICK S. HOLLIS The food material which has accumulated at the bottom during the period of stagnation is also distributed through- out the water by the overturn, thus supplying abundant food for the support of a large diatom growth, which has com- monly commenced before the time of the overturn. As the water remains in circulation until the surface water be- comes enough colder to make it less dense than that of the lower layers, the diatom and other growths become generally quite evenly distributed. One of the characteristics of the stagnant layer of water is a marked increase of color. The temperature and color at the surface, mid-depth and bottom of Lake Cochituate for a year, indicating the quies- cent state and the sprimg and autumn overturns are given on the accompanying plates. (Plates VII and VIII.) The distribution of the micro-organisms before and at the time of the autumn overturn of the water for the same year is shown by the following analyses: LAKE COCHITUATE—1896 MID-DEPTH BOTTOM SURFACE i Pe ahive | a a 2 Py } ; @ 2 | n <>) DATE a18ie!1 |x gsi ¢ lx|l al si 2 As a eae = ee = | ed a es |= ge} S) 5) Sg] &| S18] Sigel 8| &| Ble g2 2\5 a) ajee 2/8/83) ISB 2] 8) 8] a Si een Relea Sel Stee abs al rela seve ie Sa SS Ba?) @| S| 8} 6 leo] 6 OS eS aie eames — —— —|! | ERS fase tan eae eee | = | on a Oeb 20.2 | 476| 333 72] 27| 00|| 397| 293| 26) 33/ 4|| 56| 54) oO] 0} 0 Oct! 2702 4): 446) 235] 258] 7| 70|| 574) 289| 180} 29] 58|| 483] 258) 42] 3] 174 Noy. 3 | 762, 645, 34] 43) || 638) 568] 18) 27) 20|| 849] 357/ 0} 16) 428 Noy. (9s, 233) | 742 677; 26; 26| _8|| 880) 783} 36| 17/ 10}) 655] 422) 0) 28) 152 Novs16..... 1279 805) 334) 25) 114//1344'1004| 250, 40) 50)|1319/1223; 28) 0| 88 Noy. 23 ....|2016 1725 188! 17! 78|/1924'1576| 256, 17| 50/|1701/1425! 228} 18) 36 Noy. 30.....|1479 1218) 200, 47; 10) 1319/1039) 258) 7) 12//1304)1156| 84) 36| 28 13, 10 1761/1506) 228 14 Dec. 7....../1980 1633, 258) 63| 26|/1762 1502) 200 1288 : Veiga Dec. 14.....|1876 1570) 182' 104! 4|/145411288' 76! 68 0/173911524! 861| 26! 20 That the amount of water at the bottom of the lake in which these stagnation effects are marked is insignificant compared with the whole volume of the water is evident from the very slight increase of color imparted by the water of the stagnant layer to the water of the other depths at the time of the overturn. As has been stated, the surface soil has been removed from the entire area of most of the reservoirs and in these the stagnation effects and the collection of food material at the GROWTHS IN SURFACE WATER-SUPPLIES 55 bottom is very slight, although the same dissemination of organisms throughout the different depths at the time of the overturn is noted as in the case of Lake Cochituate. DIATOMACEAE In Lake Cochituate, as a result of this mixing of the diatoms at all depths by the spring and autumn overturn of the water during the time of development of the diatom growths, together with a considerable local growth of Melosira at the bottom, the average diatom growth for 1898 and the first month of 1899, during which time the autumn growth continued, was as follows: Surface, 326; mid-depth, 358; bottom, 372. The same tendency is shown in the Sudbury Reservoir to- ward a more uniform number of diatoms at all depths due to the mixing at the time of the overturn, although all the con- ditions are favorable for a normal development. The average for the year 1897 was 84 at the surface, 81 at the mid-depth and 61 at the bottom. For the period between the first of May and the first of December, 1899, the average for this source was 329 at the surface, 283 at the mid-depth and 199 at the bottom. Aside from Lake Cochituate but few of the reservoirs of the Metropolitan supply support diatom growths which are ever large enough to be seriously detrimental to the character of the water. Furthermore, while the average for the year may be so influenced by the large numbers Fhich follow the period of overturn and extend to all depths, there are many diatom growths during the year where there is, for part of the period of growth at least, a marked tendency to local development at a particular depth, in which case the forms can frequently be avoided, along with the other growths, by drawing the water froma depth at which the diatom growth does not exist. Such a growth is Asterionella, which commonly develops in largest numbers at or near the surface. I recall one in- stance of a surface growth of Asterionella amounting to about 250 stand. units per cc. in a comparatively small reser- voir 30 ft. deep in a hilly or almost mountainous district in Pennsylvania, which was entirely washed from the reservoir over the spill-way by a single heavy rain, during a period when I was studying the supply. 56 FREDERICK S. HOLLIS Most of the diatoms impart an oily or aromatic odor and taste to the water which is characteristic of the form, but generally not particularly well marked. This odor is gen- erally increased somewhat by heating. Asterionella is an exception and is characterized by a well- marked distinctive aromatic odor resembling rose-geranium leaves, which is frequently lost by heating. The forms of most importance in determining the purity of a water-supply are found among the Cyanophyceae, and Infusoria and to a lesser extent among the Chlorophyceae and Rotifera. These undoubtedly all tend to a local develop- ment during the period of maximum growth in a reservoir in which the conditions are such that a normal growth is possible. CYANOPHYCEAE Of the Cyanophyceae, Anabaena is perhaps the most com- mon and the most objectionable form, as it develops in large numbers and imparts its characteristic choky odor and un- pleasant taste to the water and the odor is much intensified by heating. It tends under normal conditions to a maximum development during the period of growth at the surface, where it collects in large numbers in areas which are moved about the surface of the reservoirs by the action of the wind. It is frequently mixed through the water by heavy winds or by the flow of a large volume of water through a reservoir, but, if in a vigorous growing condition, it tends to rise again to the surface. In the Sudbury Reservoir during a period of growth from the middle of August to the middle of September, 1897, the average was 346 at the surface, 81 at the mid-depth and 18 at the bottom, with a maximum growth of 684 at the surface. For the same source for the period of growth between the middle of May until the first of November, 1899, the average for the surface was 121, for the mid-depth 69 and for the bottom 48, with a maximum growth of 648 at the surface in August. The same is true for Framingham Reservoir No. 3, Lake Cochituate and the other sources in which it develops. The largest growth of Anabaena that has ever come to my atten- tion was one in the same Pennsylvania reservoir in which the growth of Asterionella was noted, where it showed the GROWTHS IN SURFACE WATER-SUPPLIES 57 same tendency to a maximum development at the surface. The samples were taken July 27, 1897, and showed 5,100 at the surface, 670 at the mid-depth and 123 at the bottom. Clathrocystis is another form of Cyanophyte which is quite generally distributed and causes difficulty in a supply by imparting a sweetish odor and taste suggestive of the husks of green corn to the water. Its distribution is best studied in the Hopkinton Reservoir in which it has reached large numbers in recent years. The most abundant growth occurs betwween June and November. Like Anabaena it tends to grow at the surface and to form patches. The averages for the periods of growth for the last three years and the maximum growth at the surface are as follows: Surface Mid-Depth Bottom Maximum Growth at Surface 1897 1644 645 695 3460 August 10 1898 824 246 46 2900 June 28 1899 386 15 48 2000 June 20-27 Coelosphaerium, while quite as widely distributed as Clathrocystis, is not, however, as objectionable. It is pres- ent with the growth of Clathrocystis in the Hopkinton Reser- voir and between May and November of last year showed an average of 144 at the surface, 98 at the mid-depth and 102 at the bottom. A growth in Framingham Reservoir No. 3, between May and October, 1895, showed an average of 489 at the surface, 469 at the mid-depth and 412 at the bottom. It tends to grow at the surface and to collect in masses as do the other members of this group but, on account of its more compact structure, it seems more apt to remain at a depth when carried there by the action of the wind. Aphanizomenon, which occurs as large growths only in Lake Cochituate, imparts a characteristic sweetish taste to the water which is not, however, as objectionable as that of the other Cyanophyceae already described. The growth com- mences at a depth and is first noted at the mid-depth and bottom during July. The maximum growth is at the surface, where it is very abundant in the form of flocks, and generally occurs late in November or during December. As the growth is well developed at the time of the autumn overturn it is generally quite well distributed at all depths. The growth in 1898, which was rather larger than usual, appeared at the mid-depth and bottom on June 27, reached 58 FREDERICK 8S. HOLLIS a maximum of 1385 per ce. at the surface on December 12, and continued until February 6 of the following year. The average for the period of growth was 318 at the surface, 220 at the mid-depth and 144 at the bottom. Microcystis is frequently abundant and attains a large erowth at the bottom as well as at other depths. It is, how- ever, not objectionable in the quantity in which it is found in our reservoirs. Oscillaria is observed floating in flakes attached to thin plates of mud after it has risen to the surface. Its presence has never given rise here to any objectionable condition of the water. CHLOROPHYCEAE Among the Chlorophyceae, the floating forms that are met develop maximum growths at the surface. Their presence has never caused any objectionable qualities in the water of our reservoirs. Protococcus is of very common occurrence at certain sea- sons of the year, but is rarely abundant. A growth in the Sudbury Reservoir between July 7 and October 27, 1897, amounted to an average of 53 at the surface, 24 at the mid-depth and 7 at the bottom. Gonium has at times been quite abundant at the surface of part of the Sudbury Reservoir. Spirogyra, Conferva and Draparnaldia are common as growths along the lower course of the influent streams, but they are rarely met in samples of water taken at the lower end of the reservoir. DESMIDEAB Among the Desmideae, Staurostrum is the only form that is ever found in any abundance in the main body of water of the reservoirs. Its presence has never caused trouble. Other members are common in the influent streams and de- tached shallow portions of water. The presence of Crenothrix is characteristic of the stagna- tion effects at the bottom of a reservoir and, unless washed in in large numbers from adjoining swamps, is present in the main body of water only after an overturn of the water. INFUSORIA The Infusoria are perhaps the most objectionable forms en- countered in water-supplies, both on account of the objection. Sea GROWTHS IN SURFACE WATER-SUPPLIES 59 able odor and taste imparted to the water by many of them and on account of their universal distribution and very rapid development. The more objectionable ones tend to develop in large numbers at or near the surface, but are frequently distributed through the water and, during the decline of a growth, they frequently collect near the bottom of a reser- ie MOIr. Uroglena is frequently present in large numbers between early autumn and the following summer, and imparts a strong and unpleasant oily odor and taste strongly suggestive of whale oil soap to the water. This odor is much intensified by heating. Normally, it tends to develop in greatest abund- ance at the surface. Such a normal growth is just disappear- ing from Lake Cochituate. The average number of Uroglena between May 17 and June 18 was 973 at the surface, 157 at the mid-depth and 86 at the bottom, with a maximum growth of 3800 at the surface on May 31. The largest growth at Uroglena noted in our reservoirs was in Framingham Reservoir No. 3 in 1897, at a time when the water of the reservoir was uniformly turbid as the result of work in progress on a reservoir above it on the same water shed. The growth extended to all depths from the time of its ap- pearance and lasted from May 12 to June 23, 1897. The aver- age at the surface was 2178, at the mid-depth 2288 and at the bottom 2696, with maximum growth of 4700 at the surface and mid-depth. It is not uncommon for Uroglena, when seeded into a stor- age reservoir, to develop to such an extent as to be higher in the water thus contaminated than in the original source. Synura is another form which may be expected at almost any time of the year except during the most extreme heat of summer, although it is most common in cold weather. It imparts a characteristic and unpleasant taste and odor to the water and this is intensified by heating. Like Uroglena, it is capable of increasing rapidly if seeded into a reservoir from a contaminated source. Normally it tends to develop in largest numbers near the surface, but a vigorous growth generally extends to a considerable depth. A growth in the Sudbury Reservoir between the first of April and the middle of May showed an average of 31 at the surface, 27 at the mid-depth and 1 at the bottom, with a maxi- mum growth of 134 at the surface on May 5. 60 FREDERICK 8S. HOLLIS A considerable Synura growth in Lake Cochituate in 1897 was distributed as follows: Stand. units per ce. of Synura . Taste of Water Taste of Concentrate Feb. 7 Surface 286 Synura taste Strong Synura taste Feb. 8 Surface 116 Synura taste Strong Synura taste 5 ft. 144 Synura taste Strong Synura taste 10 ft. 74 Synura taste Strong Synura taste 15 ft. 64 Synura taste Strong Synura taste 20 ft. 48 Synura taste Synura taste 25 ft. 82 Slight Synura taste Slight Synura taste 36 ft. 0 No Synura taste No Synura taste 45 ft. 10 No Synura taste No Synura taste 55 ft. 42 Slight Synura taste Synura taste Dinobryon, Glenodinium, Peridinium, Chlamydomonas and Mallomonas have been noted in considerable numbers and all impart an odor and taste to the water. The first three give it an oily odor which is increased by heating. Chlamydomonas causes an oily odor when present in moderate numbers and a disagreeable odor when very abundant. Mallomonas when very abundant imparts an odor suggestive of violets. Dinobryon and Glenodinium reach the highest numbers at the surface, but are frequently present in large numbers at lower depths, especially during the decline of the growth. Peridinium, while common at the surface, is frequently found in abundance at the lower depths. Chlamydomonas has been observed in large numbers but once in any of our reservoirs. From August, 1898, to late in the spring of the following year it was present in con- siderable numbers and, while mainly a surface growth, it was present to the extent of about 150 at the surface, mid- depth and bottom when it reached its maximum growth in March, 1899. Mallomonas is of frequent occurrence, but has never been the cause of trouble. It is a form which develops at a con- siderable depth and is brought to the surface by the circula- . tion of the water. It tends to settle back to its original posi- tion. Lake Cochituate and Whitehall Reservoir have shown the most marked growth. It was brought to the surface of one portion of Whitehall Reservoir to the extent of 2168 standard units per cc. by a wind storm in August, 1897. Four days afterwards the maximum growth of 1936 standard units per cc. was found at a depth of 10 feet. On the middle of Septem- ber it was all near the bottom. GROWTHS IN SURFACE WATER-SUPPLIES 61 ROTIFERA Rotifera are frequently quite abundant, but not often to an extent sufficient to influence the character of the water. While the largest growths are generally at the surface, it is not uncommon for them to appear first at the bottom of a reservoir. Their appearance seems often to follow a growth of Infusoria. Polyarthra, Synchaeta and Anuraea are the forms most commonly observed. It has been impossible for me, in the short space of time that I have been able to allow myself for the preparation of this paper, to make a sufficient number of averages from the occurrences of the different growths to give the results the definiteness at which I had aimed. Such as I have made are, however, selected carefully from the great mass of accumu- lated results as types, and have been worked out for periods of a considerable length of time. Many averages not here given have been prepared from the results of the other sources and have served merely to confirm the ones selected, which better represent the typical growths. They are, therefore, only types of the many hundred that might be produced in the case of any of the forms more commonly met, and will, I think, justify the following con- clusion: With reservoirs properly constructed, from which the surface soil has been removed, so that marked stagnation effects are avoided, with outlets at the surface, mid-depth, and bottom, and so arranged that an individual reservoir can be cut out of the chain in case of contamination from a growth, it is possible by watching the water through the regular examination of samples from the surface, mid-depth and bot- tom and well directed field work, as a means of tracing the development of a growth through a reservoir, to avoid almost entirely the results of such growths. Such information en- ables one to fill one reservoir from another when the water thus stored for future use is in its best condition and to supply water for consumption as free from growths as the nature of the supply permits. Even with reservoirs less carefully constructed and less fortunately situated than those of the present Metropolitan supply, much can be accomplished by such study of the sources. Laboratory of the Metropolitan Water Works, Boston, Mass., June 26, 1900. 62 GROWTHS IN SURFACE WATER-SUPPLIES EXPLANATION OF PLATES Plate V Graphic representation of the average of weekly analysis from 1895 to 1899, inclusive, to show the abundance of organisms at sur- face, mid-depth and bottom for Sudbury, Hopkinton and Ashland reservoirs. Plate VI The same for Framingham reservoirs Nos. 2 and 3 and for Lake Cochituate. Plate VII Graphie representation of yearly record of temperatures in Lake Cochituate for 1896, showing the quiescent state and the spring and autumn overturns. Plate VIII The same for color in Lake Cochituate during 1896. PLATE V ORGANISMS AT SURFACE,MID-DEPTH AND BOTTOM AVERAGE 1895-9 Standard Units per cc oct._| Nov Po. Yeprl Ay. be PLATE VI ORGANISMS AT SURFACE MID-DEPTH AND BOTTOM. AVERAGE (895-9. Standard Units per ce Sur Mid. ----- Bot. —.—-— : : 1 JAN. | FEB | MAR. | APR | MAY | JUNE | JULY | AUG. | SEPT se ae) ‘— LAKE COCHITUA ” ne PLATE VII TEMPERATURES LakE COCHITUATE 1896 BIOLOGICAL LABORATORY , BOSTON WATER WORKS. TEMPERATURE AT SURFACE = —— 2 * M1D-LEPTH —-—— : " BOTTOM ~~ L* PLATE VIII Cozors LAKE COCHITUATE 1896. BIOLOGICAL LABORATORY , B0STON WATER WORKS CozoR ar SURFACE : * Mio-bEPTH —-—-— . * BOTTOM -----~------ JAN. FEB. MAR. APR. MAY SUNE SULY AUG. SEPT. OcT. NOY. DEC. 3.00; 3.00 2.50 2.50 2.00 bese ‘ 2.00 1.50 a ; 150 10 i 400 LIMNOLOGICAL INVESTIGATIONS AT FLATHEAD LAKE, MONTANA, AND VICINITY, JULY, 1899 By MORTON J. ELROD, UNIversiTy oF MONTANA WITH NINE PLATES The University of Montana Biological Station was organ- ized in the summer of 1899, and consequently but one season’s work has been done. The organization of the work was made possible through contributions from friends in the state, con- tributions being made from individuals in Missoula, Kalispel, Butte, and other places. The object of the station is twofold: (1) to offer a place where biological investigations may be pursued during the summer months, where the collecting season is short and con- centrated, and to encourage students in their work, to offer them facilities, and to bring biological study to a higher plane in the schools; (2) te pursue systematic work along definite lines with a view of working out some scientific problems, to make collections for the University work and for the museum, and to work up the natural history resources of the state. The plan for the work was presented to the State Board of Education, which heartily approved of it. The station was placed on the same basis as a department of the Univer- sity, and so far as possible appropriation was made for its maintenance. The work of the first year was preliminary, most of the time being spent in laborious detail work, in fixing up a laboratory, looking after boats, seeking collecting sites, and in similar duties. Nevertheless, a dozen workers were gathered together, much good material was collected, and a good beginning made. The station facilities are not large, but present ample opportunity for work as a beginning. A small field laboratory has been erected, with tables for twelve students, a dark room for photography, and a store room. The boats consist of a gasoline launch capable of carrying eight people, a row boat, 64 MORTON J. ELROD and a canvass boat for use in mountain lakes and in remote regions where a boat must be transported. Microscopes, glassware, chemicals, books, and all necessary materials are taken to the field laboratory from the University. Nets after Kofoid’s plans, and also a pump for plankton, after plans by Ward, have been made. Apparatus for taking fish and insects, cameras, firearms, etc., are provided. The boats and equipment referred to can be seen in Plates XVI and XVII. During the first season very little work could be done on Flathead Lake. A number of soundings were made, and at each sounding the net was let to the bottom and hauled to the surface. Although this method was unsatisfactory, yet the results are very interesting. Surface hauls were made on sey- eral occasions. In addition to this work on Flathead Lake considerable time was given to Daphnia Pond, near the labora- tory, and described later. A day was spent at McDonald Lake in the Mission Mountains. Very little work has been done in the region, or in Alpine lakes in general in this section. Ichthyological work was car- ried on by Dr. David S. Jordan in the Yellowstone National Park, in 1889, and by Prof. W. B. Eyermann, in Montana and Wyoming, in 1891. In 1890, Prof. Edwin Linton, of Washing- ton and Jefferson College, Pennsylvania, and Dr. S. A. Forbes, of the University of Illinois, together made extensive study of the life of the waters of the Yellowstone National Park, the former having in hand the study of fish parasites, the latter of fresh water invertebrates. In 1891, Dr. Forbes and Prof. Everman spent some time im the region around and adjacent to Flathead Lake; the former again looking after fresh water invertebrates, the latter collecting fishes and seeking a suitable place for a trout hatchery. The results of Dr. Forbes’ work are given in a paper of 52 pages, with six plates, in the Bulletin of the United States Fish Commission, Vol. XI, pp. 207-258. The work of these men is all that has been done on the life of these lakes, so far as is known to the writer. The map (Plate IX)* will give an idea of the general * Map showing the section of the state north of Missoula to the boundary line, and from the main chain of the Rocky Mountains west to the Idaho boundary line. Only a few of the smaller lakes are included. The rivers and streams are not ac- curately drawn, but are to the best of our present knowledge. Few of the moun- tain ranges are indicated. Each water course is a canyon, usually narrow, between two ranges of hills or mountains. Few wagon roads have been made through the canyons, but of those existing only two or three are located. Most of the streams, many of the lakes, and all of the peaks are inaccessible except on foot or by pack train. fol PLATE IX € aval? RIT = ae DEER LODGE \ Cp‘ TY LIMNOLOGICAL INVESTIGATIONS 65 outline and shape of Flathead Lake, the streams flowing into the lake, the outlet, routes of travel, and other points of in- formation. A brief description of the lake, with its geologi- cal history; will be of importance in taking up the study of the life found. The geological description here given is furnished by Prof. Fred D. Smith, of the University of Mon tana. (Cf. Plates X, XI, XIL.) “The lake occupies the lowest portion of an immense valley that reaches from the Jocko Mountains, a low range be- tween the Jocko River and Mission Valley, northward across the British Columbia line into the latter country, a distance of over one hundred miles. It is but the remnant of a lake that in Tertiary times occupied this valley through- out its whole extent. The great level plains on either end of the lake are the beds of sediment deposited in the former lake, and show by the character of their soils that the lake was a large and quiet body of water. The plain on the south- ern end of the present lake is about thirty-five miles long. On the northern end of the lake the plain extends a distance of sixty miles to the border of the United States and into the British possessions. “The valley, as well as the lake throughout much of its length, is bordered on the eastern side by the Mission Moun- tains, a range which rises abruptly from the plain to a height of 10,000 ft. These mountains, with a very steep western slope, have their summits within relatively short distances from the valley, and consequently the streams therefrom are not large nor of great volume in discharge. The peaks of the range rise bare and steep. The range appears to terminate as such at a point near the upper end of the lake where the Swan or Big Fork River changes its course from northward to west and southwestward, to flow into Flathead Lake. “Mission Valley, Flathead Lake, and Flathead Valley ex- tend about a hundred miles from north to south; Flathead Lake separating Mission Valley on the south and Flathead Valley on the north. Perhaps the most interesting feature of the region represented is its drainage. The drainage from Flathead Valley is through the Flathead River. This has three great tributaries, the South, Middle, and North Forks. The latter, only, is a real factor in the drainage of the valley. The Flathead River flows into Flathead Lake from the north, as does also the Swan River. These together materially in- 5 66 MORTON J. ELROD crease the size of the lake in the spring time. The outlet of the lake is the Pend d’Oreille River, also called Flathead, which flows out of the lake at the southern extremity. Fol- lowing a circuitous route in a south-westerly direction, it receives the streams that cross the southern portion of the valley transversely, and eventually unites with the Missoula River to form Clarke’s Fork of the Columbia. Considered thus, Flathead Lake appears as an enlargement of Flathead River, and as one element in the drainage system. “The Mission Mountains were made by an immense fault, having the general direction of north by south. The moun- tains were raised, while the corresponding strata on the western edge of the fault were depressed, thus producing the usual basin for the immense lake which afterwards filled it. Possibly the lake was not a part of a drainage system, as the present lake is, but acted as a large reservoir. When the lake was drained, probably through a passage to the north,* there was no large amount of run-off from any exten- sive drainage system to be carried away. The small streams that came from the lower part of the Mission Mountains cut small water courses directly across the beds to the west in parallel directions. Flathead Lake receded to the lowest parts of the depression in the great valley, which were ap- proximately the central portions. The lake may have ocecu- pied different levels in its present position, though it has probably never been high enough to receive any of the drain- age of the lower Mission Mountains, owing to a larger em- bankment along its southern end. This ridge may be of morainal origin, and probably was, since it is higher than the surrounding plains on either side, and no evidence has been observed of higher levels of the sedimentary lakes. “When in its new position the lake, receiving considerable inflow from the north, began to find an outlet across the beds in a southwesterly direction towards the Missoula River. Whether this was caused by a damming of the streams on the north by glaciers or by elevation of the country is not plain at present. The carving of what is now the Pend d’Oreille River canyon probably was rapid, and the lower plains on the north of the present lake were uncovered, thus making the fertile areas south of the city of Kalispel. The * Later research indicates that the passage was out of the western bay, possibly near Dayton. LIMNOLOGICAL INVESTIGATIONS 67 Flathead River in its present condition is but a remnant of the lake which extended over these areas. “This Flathead River winds its way in a very circuitous path across the plains, and has a total length of about thirty- five miles, while the distance as measured by a straight line is but fifteen miles. In general its width is from three hun- dred to six hundred feet, and its depth is over twenty feet in all places, and often reaches seventy-five feet. For these reasons it may be considered but an arm of the lake, since its level is the level of the lake except for sufficient fall to cause the waters of the tributaries to flow to the lake. On account of the very sluggish nature of the current of this river the erosion of the banks is slight, while the deposition in the bottom and at the mouth of the river is rapid. “The northern end of the lake around the mouth of the river is apparently composed of sediments deposited as a large delta in the manner mentioned. The course of the river is plainly traced into the lake for some distance by the delta thus formed, which for a distance of from one-fourth to one-half a mile from the shore is sufficiently high to be covered by vegetation, and in some places by shrubbery. Be- neath the surface of the water the formation is discernible for a long distance farther into the lake. “At the end of the Swan River Valley near the location of the Biological Station are to be seen many rounded hills which are probably morainal in origin. On the slopes of the Mission Mountains that form the termination of this range are found many evidences of glacial action in form of smoothed rocks, post-glacial gorges and stream courses, glacial scratches, ete., while the glacial origin of the ridge at the foot of the lake has already been suggested. There is no doubt but that glacial agencies have materially affected the history of the lake both in its present and in its older form. To what extent moraines may affect the contour of the lake bottom can only be surmised, but as they are apparent on the beds of the older lake it is to be expected that they may be found on the bed of the present lake.” The outlet is called by some Pend d’Oreille River, by others Flathead River. Some consider Flathead River to extend from its source to the lake, then from the lake to the Missoula River. Others give the name Pend d’Oreille to the stream from Flathead Lake to the Missoula River. The river formed 68 MORTON J. ELROD by the junction of the Missoula and Pend d’Oreille is called Clarke’s Fork of the Columbia. The present outlet of Flathead Lake is of recent origin. The river for several miles near the lake is swift and rocky, a series of rapids alternating with more quiet water. About a mile from the lake there is a large bank of clay through which the river has cut. The clay is continuous with, and ap- parently a part of, the moraine mentioned. At the river bank it has been cut and eroded by the wind and rain. The bank is abrupt and steep, the clay clinging together so as to form cliffs, some ending in sharp pinnacles. Below the clay is the bed rock, similar to that found at different places around the lake. The river has done some cutting through the solid rock bed, but not much. At one place the channel is partially dammed by a large rock in the center of the river. Above and below this place the river is a beautiful sheet of foam, with several small falls. It is as beautiful a rapid as one usually sees. In my estimation it is superior to the rapid above the first falls in the Yellowstone. While not so large, it impressed me more deeply than did the rapids below Ni- agara. Several cases have been reported of people who were overcome by the sight close to the water’s edge and had to be carried away. Plate XIII shows the rapids as seen from the hillside a couple of hundred feet above the water. This is a great fishing resort for the Indians on the reservation, and one seldom visits the place without seeing several tepees on the bank some place near. The osprey is as industrious as the Indian, and is seldom absent from the scene when one visits the rapids. The banks of the lake do not afford as much shelter for invertebrate life as would at first seem apparent. The south- ern third, cut off by the islands, is shallow, nowhere of greater depth than twenty feet. The eastern slope of this bay, formed by the peninsula projecting from the Mission Mountains, is very marshy, with muddy bottom. Rushes and weeds grow abundantly, offering an excellent harbor for smaller life. This is the largest marshy region around the lake. Between the mouth of Flathead River and the mouth of Swan River, along the northern shore, is another marsh in the spring, of peculiar nature. At the water’s edge is an embankment of a more or less rocky nature. North of this embankment is a shallow marsh, a couple of miles long and a quarter to a LIMNOLOGICAL INVESTIGATIONS 69 half mile wide. When the lake rises, as it does in the spring, from ten to twelve feet, the water flows over the embank- ment, and into the low land. As the lake recedes the im- prisoned waters cannot escape, and offer a fine breeding place for mosquitoes for some time, until the waters evaporate or filter through the soil to the lake again. Most of the remain- ing banks are rocky, precipitous at the water’s edge, with or without a gravelly beach. The bottom generally is re- ported to be rocky, with little mud. This report comes from the captain of the boat Klondyke, who has anchored all over the lake; his experience on the lake extends over a period of many years. Compared with the size of the lake the swampy country is small. From this it would appear that the breeding grounds for most of the fish must be in regions distant from the lake, causing long migration periods. This is made more apparent from the fact that fish are rarely caught any place in the lake except at or near the streams entering the lake, or at the outlet. Flathead Lake is populariy supposed to be very deep. I was told it was 1,500 ft. deep in places. During the summer of 1899 some twenty soundings were made in the lake and rivers. The greatest depth obtained was 280 ft. The location of this may be found by referring to the map. Eugene Hodge, captain of the Klondyke, states that nowhere is the water deeper than this sounding. During the season of 1900 other and more numerous soundings will be made. McGovern Bay, on the northern end of the lake, is about seventy feet at the deepest. Flathead River has filled in a large amount of sediment. East of the mouth of Flathead River the drop in depth is sudden from the river bar. The deepest portion of the lake is off shore on the east side, next the Mission Mountains. In high water a great deal of land at both ends of the lake is covered. If the depth of the lake should be lessened by ten feet, thousands of acres at the lower end would be uncovered. The annual rise and fall of the lake is from ten to fourteen feet, but it has risen as much as nine- teen feet in a season. The lake acts as a huge reservoir for water storage, but overflows much land almost every year when it is at the highest. The amount of water flowing into the lake and out of the lake annually has not as yet been de- termined. Life in Flathead is scarce. Although some species are taken 70 MORTON J. ELROD in great abundance, the cold clear waters, with rocky bot- tom and banks and with few marshes, make life scarce as compared with similar bodies of water located in warmer climates at lower altitudes. The first collecting done with the net was on July 22, the last August 11. The method employed was to let the net to the bottom and slowly bring it to the surface. This was not satisfactory, but was the best that could be done at the time. The material from each haul was placed in a vial and numbered, the data being recorded. Twenty-one numbers were taken at Flathead Lake, an additionel number at Me- Donald Lake. As will be seen from the data subjoined these collections were made at different parts of the lake, and represent the life of the lake at this season fairly well. It is to be regretted that material could not be taken both earlier and later in the season, but this will have to await further developments. The record of collections and material, with data, is as follows: No. 1. July 25, 11:00 a. m., bright sunshine. Swan River, opposite Sliter’s house, near the Station. Contents, sand. No. 2. July 25, 11:20 a. ms. Mouth of Swan River opposite club house. Contents, sand. No. 3. Bottle lost, not examined. No. 4. July 26, a. mM. Bay in front of club house, in the waters from the Swan River. Contents, nothing that could be determined. No. 5. July 26, 10:00 a. um. Opposite first bluff below club house, where the waters from the river have become quiet; depth, 60 ft. Contents, a few Hpischura nevadensis Lilljeborg, Diaptomus ashlandi Marsh quite numerous, about as many Cyclops pulchellus Koch, and a few Cladocera. No. 6. July 26, 10:30 4. m. Between club house and mouth of Flathead River, nearly a mile from shore and perhaps a mile and a half from the river; depth, 96 feet. Contents, Cyclops pulchellus Koch made up the bulk of the material taken, Diaptomus ashlandi Marsh was rather abundant, and a few Daphnids. No. 7. July 27, a.m. Flathead River, opposite Holt, which is about three miles from the mouth; depth, 56 feet. Con- tents, a few Diaptomi. No. 8. July 27. Half mile below No. 7. A few each of Daphnia and Diaptomus. LIMNOLOGICAL INVESTIGATIONS a! No. 9. Mouth of Flathead River, same date; depth, 18 feet. Contents, nothing but sand. No. 10. July 26. Lake, east of the mouth of the Flathead River; depth, 10 feet. This was in the northern shallow end of the lake, but not on the sandbar which receives the waters from the river. Contents, Cyclops pulchellus Koch, Diaptomus minutus Lilljeborg, in about equal quantities. No. 11. Bottle lost. No. 12. July 26. Lake, one-half mile east of club house; depth, 10 feet. Conditions similar to those in No. 10. Con- tents, a few Diaptomi, with an occasional Daphnia thorata Forbes. No. 13. Lake near rocks by club house; depth, 15 feet. This is in the waters of the Swan River. Contents, a few Cyclops pulchellus Koch. No. 14. Bar at the mouth of Flathead River. Contents, nothing. No. 15. Lake between Flathead River and the club house; depth, 40 feet. Contents, Diaptomus ashlandi Marsh, Cyclops pulchellus Koch, with three or four specimens of a larger form of Cyclops, 2 mm. long. No. 16. August 11. Lake about six miles below Chap- man’s, east side, about three miles from shore, not far from midway of the length; depth, 280 feet. Diaptomus minutus was found in large quantity, and Cyclops pulchellus Koch in somewhat smaller amount. No. 17. August 11. Near the islands on the north, in the “channel” used by the steamboats; depth, 15 feet. Not ex- amined for species, but no doubt similar to No. 16. No. 18. August 11. In shallow water below or south of the islands; depth, 17 feet. Not examined for species. No. 19. August 11. Lower end of the lake, about one mile north of the islands, opposite the point of land on the west, and the middle of the flat-topped mountain on the east; depth, 167 feet. Contents, Cyclops pulchellus Koch, Diaptomus, probably ashlandi, though slightly smaller, and one specimen of Epischura nevadensis Lilljeborg. No. 20. August 11.. Near No. 19, but at a depth of 75 feet. Contents, Cyclops pulchellus Koch in largest quantity, Diaptomus in smaller quantity. No. 21. August 11. Several bottles of skimmings from the surface of Flathead Lake at different places. On this date G2 MORTON J. ELROD a round trip was made to the foot of the lake. At different places skimmings were taken with the net. Cyclops pulchellus was taken in very large quantity, and was very noticeable. While at Chapman’s, on the east side, for wood, a bottle was shown him, which rather startled him when he con-— sidered he was drinking from the lake. The only comment made was that there were a good many people drinking from the lake, and he was not alone. To dip up a tin cup full of water was to take numbers of them. As the day was bright, in the middle of August, this is rather surprising, as they generally stay down during sunshine. Moreover, Forbes re- ported Cyclops as very scarce in his collecting. No. 22. August 18. Collection made at McDonald Lake, as recorded under description of that lake. The list taken from Flathead Lake is not large, and is as follows: Diaptomus ashlandi Marsh. Cyclops pulchellus Koch. Epischura nevadensis Lilljeborg. Diaptomus minutus Lilljeborg. Daphnia thorata Forbes. A few Cladocera. Some young that could not be determined definitely. Of these Cyclops pulchellus was exceedingly abundant, taken at nearly every point on the lake where collections were made. Daphnia thorata was scarce, which is surprising from the fact that Forbes relates that in his haulings with the surface net in late September, 1891, this species made prob- ably from four-fifths to nine-tenths of each haul. He also records that Daphnia pulex was not seen at all, though com- mon in Yellowstone Lake. Daphnia pulexr was taken by thousands in Daphnia Pond, near the Station, as recorded in description of work in this pond. He also records Epis- chura nevadensis, var. columbiae as very common, but with us it was scarce. It therefore seems that the Entomostracan life is undergoing great changes, which will offer good field for investigation. It seems peculiar that such complete changes should be made in the waters of a lake of this size as indicated by this comparison. The absence of Daphnia pulexr from Flathead Lake, and its abundance in Daphnia Pond, which is but a few rods from the lake, suggests either that this species does not like cold LIMNOLOGICAL INVESTIGATIONS (3 water, or else that it is preyed upon by fish. Since it is com- mon in Yellowstone Lake, neither of these explanations would be satisfactory. The absence of Daphnia pulex and the great abundance of Cyclops pulchellus, as noted, need explanation. McDonald Lake of the Mission Mountains lies at the foot of McDonald Peak on the northwest. It is about eleven miles from St. Ignatius Mission, and about fifteen miles due north of Sin-yale-a-min Lake. Sin-yale-a-min Lake is at the foot of Sin-yale-a-min Mountain, the last on the range south next the Jocko River, which river cuts the range in two. Mc- Donald Lake, like Sin-yale-a-‘min Lake, is hemmed in on all sides except the west by mountains, but at McDonald the mountains are tall, rugged, and very picturesque. The lake was named back in the sixties, and, according to priority, the name McDonald should easily displace the same name given to Terry Lake, above Kalispel. McDonald Lake is a beautiful spot. Seldom will one find such a combination of grand mountain peaks with the quiet serenity of the water. The sun sinking in the west at the close of the long days of summer gilds the peaks with tints of surpassing beauty. Campers on the banks of the lake have seen goats on the crags above, though at present they are comparatively scarce so close to the haunts becoming frequented by man. The banks of this lake have been a resort for the Indians and white men of the region for many years. There is but a small place at the western end where camping is possible, and the banks for the remainder are abrupt, steep, and rocky, but the small grassy spot, with the peaks in the immediate foreground, is a place frequented often. Of course the usual stories are told about the great depth of the lake, and up to the time of our visit no one had any idea of the real depth, but it was said to be “bottomless.” The valley enclosed by the peaks, in which the lake now is, has been carved out by a glacier, the remnant of which yet exists on the slopes of the peak in plain sight from almost any place on the lake. The rocks along the sides have been ground smooth, and show plainly the marks of the ice. At the outlet of the canon a large moraine has been made. The water in times past has evidently been much deeper than at present, and at the upper end what is now a wooded valley was covered with water and was a part of the lake. 74 MORTON J. ELROD The lake is about a mile and a quarter long, with an aver- age width of less than a quarter of a mile. On either side the mountains come abruptly to the water, as may be seen by the illustration. At the upper end there is an unexplored small valley, abundantly wooded with large arbor vita trees and with fir, birch, and small trees of other species. The inlet divides above the lake, one branch receiving the water from the glacier visible, the other bringing the water from the amphitheatre toward the east, and has for drainage not only the peaks visible, but also the eastern slope of Me- Donald Peak. (Pl. XIV.) The bottom of the lake slopes gently (Pl. XV), showing that the lake has apparently filled up a great deal. The depth from end to end is nearly uniform, the greatest being sixty-eight feet. The lower end is shallow, the outlet being crossed by a ford, hub deep at the time of the examination, late in July. There is considerable shallow water, and the bottom is of mud of a reddish color, apparently from the decomposition of the soft rock on the north. At a point near the middle a ledge of rocks projects from either side, making the lake at this point quite narrow. The rocks are precipitous, and the water a few feet from the rocks is deep. These rocks are worn smooth by glaciation, and show deep and numerous glacial scratches. On the north, to the left in the illustration, the rocks are precipitous for about 2,000 feet. Four waterfalls, with small streams, tumble over the rocks, the water disappearing in the loose talus at the base long before it reaches the lake. The southern slope is not so abrupt, large masses of loose talus, with large boulders, lining the water’s edge, making a loose and spongy surface for the retention of moisture. Life in and around the lake is not abundant. Frogs and snakes are practically absent, but one of the former being seen, none of the latter. On the rocks at the water’s edge, altitude 3,300 feet, several pika, Lagomys princeps, were killed. This is the lowest altitude known to the writer at which these peculiar mammals have been killed. The banks are so steep and rough that it is all but impossible to climb along, almost an entire afternoon being spent in getting from one end to the other, a few hundred feet from the water’s edge. If explored it is very likely the upper end will show a possibility of greatly increasing the surface by increasing the depth. LIMNOLOGILAL INVESYVIGATIONS 6) On the northern side the timber is not so dense, owing to the nature of the rocks, which are steep and allow poor foot- hold for timber. On the mountain above the precipitous rocks the timber is quite heavy, largely of yellow pine and fir. The southern bank is well wooded, and the canyon at the head of the lake is densely wooded, through which there does not appear to be an entrance made by road or trail. At the outlet and along the moraine near the lake there is fine timber, some of which has been cut for rails and lumber. Everywhere there is much underbrush, making progress difficult. The road to the lake is good, and there is considerable travel over it in the summer time, as the lake is a ‘great resort for the Indians and others, who visit the reservation on account of the excellent fishing and beautiful scenery. There is no drift around the shores, most of the drift having lodged in the outlet where there is quite a jam. An ascent of the mountain, and conversation with men from the United States Geological Survey has given a com- prehensive idea of the drainage system. The upper slopes of the mountains are bare. Most of them have been partially covered on the higher surfaces with black pine, which has been killed off by fire. McDonald Peak is double, the western peak being perhaps a thousand feet lower than the eastern. The two are con- nected by a ridge with a depression in its middle. To pass from the western peak to the eastern is to descend over rock for a thousand feet, then up about two thousand. The western peak is easy of ascent, the last fifteen hundred feet requiring about four and a half hours, however. But to ascend the high summit from this peak appears difficult, though by taking the snow it is no doubt possible. So far the main peak has not been ascended from the west.* The main peak has three or four spurs projecting in differ- ent directions, behind which the snow lies in deep drifts, making ice, and remaining the year through. There is little snow on the western peak, and its importance as a snow holder lies in the fact that it permits the snow blowing from the valley in the west to pile up between it and the main peak, thus making the glacier visible from almost every part of the valley. These spurs make such protection that in * Since writing the above I am told ascent has been made this way, along the edge of the snow. Three Indians are said to have gone up and returned in safety. 76 MORTON J. BLROD three different places on the heights of this mountain the snow piles in drifts, which never melt, making three large glaciers. One of these, the one seen from the lake, is shown in the illustration, the others lying behind the spurs. The waters from these three snow masses all flow into McDonald Lake. The supply is therefore abundant and never failing. Moreover, the peaks to the north of McDonald Peak, and to the north of the lake, give much of their waters to the lake. Post Creek, the outlet of the lake, at a point some twelve miles from the lake, lower in altitude by a thousand feet, with considerable loss through irrigation, carried 473 second feet of water on the 30th of June, 1900. The microscopical life of the lake will no doubt prove interesting when it is worked up, as will be the case of most of these mountain lakes. The collecting net revealed an abundance of Diaptomus ashlandi, and the female of another form a little smaller. These were taken August 18, the net being let down to the bottom, 67 feet. D. ashlandi was abundant, being quite conspicuous on account of its red color. The steep and rocky talus along the lake produces a new species of land shell, named by Pilsbry, Pyramidula elrodi. Description of this shell is to be found in Nautilus, Vol. XIV., p. 40. About forty were secured, all dead. The dead shells are a beautiful white, their color against the dark brown or lichen colored sandstone making them very con- spicuous objects. The shells were scattered among the talus at the base of the cliffs of the mountain, and though they were conspicuous it required considerable effort to secure the few taken. Diligent search failed to reveal live speci- mens, but later search may serve to find them.* In the waters of the lake Limnaea emarginata Say is quite abundant. It apears to be of a variety distinct from any de- scribed, and for it the varietal name montana has been sug- gested. The animals cling to the rocks along the sides and bottom of the lake, seldom found away from the rocks. A few Physas were found, but they were scarce. It was sur- prising not to find a single Planorbis in the lake. Pyramidula strigosa Gld., var. coopert W. G. B., and P. solitaria Say were found abundantly in the damp woods along the lake and creek. It is interesting to note that a large series was secured which had evidently been killed by squirrels, as * Several dozen have since been found. LIMNOLOGICAL INVESTIGATIONS 77 each had a hole gnawed in the shell. These shells alive showed very strikingly the idea of protection, as it required the most careful search to find them, and repeatedly they were overlooked by the person in front and seen by the one behind. Their home is in the damp brushy woods, and to secure the series taken resulted in scratched hands and faces and torn clothes, not to speak of the discomfort of crawling among the brush on hands and knees, with the digging among the debris of old logs necessary to find them. Altogether but five species of shells were found, rather a low number considering the size of the lake and the country. During the summer of 1900 a stay of ten days is planned for McDonald Lake. It is hoped to find live species of the new shells. Further study of the Entomostraca will be made on the lake, with pumping apparatus. The adjacent country will be searched for birds, and alpine forms collected. Daphnia Pond, so-called on account of the great numbers of Daphnia pulex found in it, is a small pond of some ten to fifteen acres. It is about a mile and a half from the Station, alongside the regular wagon road, and only about a half mile from the lake, but at a little higher altitude. This pond is no doubt of glacial origin, as the entire northern end of the Mission Range has been overrun by glaciers, leaving many evi- dences behind. In the center the water is about twenty feet deep, but for the most part the pond is shallow and over- grown with rank vegetation, offering an excellent harbor for smaller forms of life. No fish have as yet gotten into this ‘ pond, and consequently the invertebrate fauna is not affected by them, and has few enemies. It is a typical place to study some of the forms of life found therein, living as they do under very favorable conditions. The varied and abundant life in this small pond is in strange and striking contrast to the limited quantity and paucity of species in the large lake, so short a distance away. The most abundant Entomostracan forms were Diaptomus lintoni Forbes, described from specimens taken in the lakes and pools of Yellowstone Park, and Daphnia pulex, so abund- ant that the water appeared of a dirty red color. Numbers of half-grown individuals were found with the adults. In the open water they were taken by the tablespoonful with an ordinary insect net. Nowhere have I ever seen anything so abundant as Daphnia puler in Daphnia Pond. Swimming 78 MORTON J. ELROD among the pond lilies, and keeping out of the open water might be seen a large species of Gammarus, an inch in length when expanded. A few Cyclops pulchellus were found among the more abundant species. Shells are numerous in specimens though not in species Planorbis trivolvis Say is the most abundant. This widely distributed species was taken in all sizes from small to fully grown. WNSphaerium partumenium Say was found among the dense vegetation, and was taken in considerable quantity. Physa ampullacea Gld. (possibly heterostropha Say) was not uncommon. Along the banks of the large lake the land form, Pyramidula strigosa Gld., var. cooperi W. G. B., was found. At the lower end of Flathead Lake, in the fine sand along the river bank, were found Planorbis parva Say, while in the sands of the lake were fragments of the bivalve, Vargari- tana margaritifera L. In insects there is likewise great abundance in Daphnia Pond. Dragon-flies were noted most especially. The first week in August, 1899, Aeschna constricta Say was exceedingly abundant. Hundreds were flying in the air, and wherever Odonata were found flying mosquitoes were rare. The exuviae of this species were taken in quantity from the rushes, cattails, tall grass and weeds. The exuviae had the characteristic living attitude, the feet firmly clasping the stalk of the plant. They were usually found a foot or two above the water, but it was not uncommon to find them even three or four feet above water, the insect having crawled this distance before transforming into the adult. Only a few larvae could be found, showing that the transformation was practically completed at this date for the species. The next largest was Libellula pulchella Drury. These were also on the wing in numbers the first week in August. During the last two weeks in July Lestes wngwiculata Hag. were emerging in great numbers. They are at first very feeble on the wing, lacking in color, with soft flabby bodies. While no birds were actually seen eating dragon-flies the presence of many king-birds, Tyrannus tyrannus, was a pretty good indication that these birds were seeking such insects for food. Other dragon-flies taken are as follows: A few Lestes disjuncta were taken. EHnallagma calverti Morse was on the wing in the middle of July in abundance. EHnallagma prae- LIMNOLOGICAL INVESTIGATIONS TD varum Hag. was taken, thus extending the distribution of this species. It is now reported only from Louisiana, Kansas, and Montana. Sympetrum scotica Donoy. was rather abun- dant, as also Sympetrum rubicundula Say, var. assimilata Uhler. Many larvae of different species were taken, but all have not as yet been determined. Case-worms were found in considerable abundance. One species builds the cases out of leaves and the stalks of the green vegetation. Leeches, water-beetles, dipterous larvae, water-bugs, and worms add to the list collected and yet unworked. Daphnia Pond is near the field laboratory,and presents good opportunity for work. Farther along the road is a second pond, which will present as good a field. Neither of these contains fish, and both teem with life in the summer time. The region near Kalispel has many lakes awaiting study. Swan Lake, about eight miles from the Station, has been un- worked save for a few hauls made by Forbes. Following up ’ the river which enters Swan Lake to the divide and down the Clearwater and the Big Blackfoot to Missoula, a distance of a hundred and twenty-five miles, one passes a dozen to fifteen lakes of different sizes which have been as yet untouched. The northern end of the state has Terry or McDonald Lake and St. Mary’s Lake, both of good size, and neither of which has been worked. The opportunities offered for work in Mon- tana are great, but difficulties and distances are also great. As but a small portion of the time during the summer of 1889 could be devoted to this work, and during this time many pressing things engaged the attention, it is not surprising if there is much disappointment at the comparatively meager results. But the way is opened, the field partially disclosed, and a trail cut through the apparently impassable wilder- ness. Each succeeding pack train will make the trail plainer and meanwhile the facilities for taking the train in and get- ting material out will be better. Moreover, it is hoped the numbers composing the pack trains will increase. More than any other one thing the naturalist working in Montana needs kindred spirits to rub up against for mutual aid, to brush away the cobwebs that accumulate, and to ask stimulating and difficult questions, even though the answers may require years of work. More work, and more valuable work, will be done in succeeding years. 80 LIMNOLOGICAL INVESTIGATIONS EXPLANATION OF PLATES Plate X Mouth of Swan River, and Flathead Lake. In the distance, to the right, about three miles off, may be seen the bar at the mouth of Flathead River. Cabinet Mountains in the distance. View is south- west. Plate XI A bit of beach at Flathead Lake, showing characteristic shore, vegetation, and drift. Plate XII A Lower end of Flathead Lake, from summit of moraine, showing islands in the distance. In the foreground to the left is the outlet of the Pend d’Oreille River. The islands are about seven miles out from the shore. The view is north.—Photograph by Chas. Emsley. B Mission Mountains, from Crow Creek, after a storm. The high peak in the center is McDonald. The view is almost directly due east. The distance is about eighteen or twenty miles. Plate XIII Rapids in the Pend d’Oreille River, near the lake outlet, Flathead Indian Reservation. View is northwest.—From Photograph by M. J. Elrod. Plate XIV McDonald Lake, Mission Mountains, Montana, from the outlet. McDonald Peak is on the right. On the left bank in the picture was found the new shell Pyramidula elrodi Pils. View is east. Plate XV Outline map of Lake McDonald, showing contour, lines of depth, and geological features referred to in text. Plate XVI Canvas boat and plankton outfit of Montana Biological Station at Swan Lake, Montana, August, 1900. At the outlet of the lake looking into Swan River. Swan Mountains in the distance to the right. Plate XVII Launch Missoula and rowboat Culex of the University of Mon- tana Biological Station in Swan River harbor, Flathead Lake. Plank- ton equipment, net, pump, hose, reel, etc., on the shore nearby. PLATE X * c~ PLATE XI PLATE XII ee Rees oer lie —_— so mae” - a % 7 ; PLATE XIII 9 eo th 8S aaa PLATE XIV PLATE XV . tae MS Donald Cedar ! Clacrated Rocks Glaciated Rocks a FROM itedhere Alt.2300 dete, iran ae iia re sale rah al eid a ay) 7 ie Wega eh r a> vy 4 7 . f 4 be ¢ : 1 ry = 7 : = My : - nue Pee H ' ‘ ¥ . es ul a * al - : ‘ ‘ a PLATE XVI “ AN ADDITION TO THE PARASITES OF THE HUMAN EAR By ROSCOE POUND WITH ONE PLATE The list of Fungi which have been reported as parasites in the human ear is somewhat large. The number of species which are well identified is much smaller. The confusion long prevalent in the groups to which the ear-parasites be- long, recently abated in the Black Moulds by A. Fischer, but still in force in the Imperfect Fungi, and the fact that otolo- gists have not been much concerned with mycology nor always able to command the assistance of expert mycologists, have united with the inherent difficulties due to the effect of situs on the forms themselves to produce errors in determina- tion, uncertainty, and even controversy. The greater number of the species reported and the greater number of the well identified and authenticated species belong to the Aspergil- laceae. Next come the Mucoraceae. The Protoascineae and Pezizineae contribute one each. In addition there are several Imperfect Fungi, some of which, however, seem to owe their place upon the list to ubiquity rather than to any special adaptation to the habitat. It seems probable that almost any of the commoner pantogenous Imperfect Fungi is liable to be added to this portion of the list at any time. The following species appear to constitute the fungus flora of the human ear: Mucoraceae. Mucor mucedo U. M. racemosus Fres. M. corymbifer F. Cohn. Ascophora mucedo Tode (Rhizopus nigricans Ehrb.). Doubtful and less known species: Thamnidium elegans Lk. (Ascophora elegans). It has been suspected, with good reason, that the forms 6 82 ROSCOE POUND referred to 7. elegans were merely Mucor mucedo with sporangiola. Mucor septatus Bezold, according to A. Fischer, is probably to be referred to M. racemosus. M. rhizoformis Lichth. Endomycetaceae. Bargellinia monospora Borzi. This curious fungus, placed by Saccardo in the some- what heterogeneous group of Gymnoascaceae, is doubt- fully referred to the Endomycetaceae by Schroeter in the Pflanzenfamilien. Aspergillaceae. Aspergillus glaucus (L.) Lk. (A. herbariorwm (L.) E. Fisch.). . repens (Corda) Sacc. . fumigatus (Fres.) De Bary. . malignus (Lindt.) E. Fisch. (Eurotium malignum Lindt.). . flavus Lk. . virens Lk. . nidulans (Eidam) Wint. (Sterigmatocystis nidulans). . niger Van Tiegh. (Sterigmatocystis nigra). Penicillium crustaceum (U.) Fr. “ P. minimum Siebenmann. Doubtful or less known species: Aspergillus flavescens Wred. Believed to belong to A. flavus. A. hageni Hallier. Otomyces hageni Hallier. . microsporus Boke. . nigrescens Robin. . nigricans Wred. (1868). . nigricans Cooke (1885). Cooke says his A. nigricans appears to be distinct from A. nigrescens, but seems to consider it the same as A. nigricans of Wreden. Siebenmann refers A. nigrescens to A. fumigatus and A. nigricans Wred. to A. niger. Cooke’s figure (Journ. Queckett Micr. Club, II, 2: t. 9 f. 3) pre- cludes identification with A. niger. Cattaneo refers A. nigricans, to A. nigrescens, which would then mean, prob- ably, A. fumigatus. A. noelting Hallier. A. ramosus Hallier. ba» bf fb bf bf fe bo fe be fe ADDITION TO PARASITES OF HUMAN BAR 83 A. rubens Green. Believed to be A. nidulans. Sterigmatocystis antacustica Cram. Supposed to be Asper- gillus niger. Otomyces purpureus Woronin is thought to belong to A. nidulans. Mollisiaceae. Mollisia auriculae (Garoy.) Sace. Peziza auriculae Garov. Fungi Imperfecti. | Alysidium rufescens (Fres.) Torula rufescens Fres. Oospora rufescens Sacc. Verticillium graphii Bezold. Acrostalagmus parasiticus Hallier. Stachylidium sp. Hallier. These seem likely to prove the same. Trichothecium roseum Lk. Stemphylium polymorphum Bon. Graphium penicillioides Corda. Coremium bicolor (Web.) Pound & Clements. Stysanus stemonites (Pers.) Corda. Spores of one of the Ustilagineae (smuts) are also reported as found germinating in an infested ear, which is not to be wondered at in view of the ubiquity of these spores. To the foregoing list we now have to add NSterigmatocystis candida. The Mucoraceae enumerated include the three commonest of the Black Moulds. They are to be found on all manner of organic substances throughout the world, and their occur- rence as ear-parasites is doubtless partially due to that fact. But some connection has been suggested between the growth of Mucor racemosus in the ear and cases of diabetes, which is rendered not improbable by the yeast-like mode of growth of this fungus and its power of acting as a ferment. WM. corymbifer has been found to be pathogenic in other connec- tions. The nature and position of Bargellinia monospora are doubt- ful. It is placed provisionally in a small group of Ascomy- cetes or sac-fungi in which no spore-fruit is developed and the ascus is of a very primitive type. But it is not certain that Bargellinia is an Ascomycete at all. The so-called one- 84 ROSCOE POUND spored ascus may prove to be some form of conidial fructifi- cation. Of the Aspergillaceae named, the ten listed as doubtful or less known are reported as ear-parasites only. But they require further study, and some at least are believed by com- petent authority to be identical with other species of more general occurrence. The type of this group, Aspergillus glaucus, is the ordinary herbarium mould found everywhere on all manner of organic substances. It occurs in the ear in the conidial stage in which the ends of certain fertile hyphae swell up and produce chains of asexual spores. The stage in which small, yellow, sexual spore-fruits are produced, visible to the naked eye, was long considered a distinct species, called Eurotium herbariorum, and by some the plant is now known as Aspergillus herbariorum. The rules of nomenclature adopted by American botanists, however, seem to justify the retention of the well known name A. glaucus. It has been suggested that the forms described under the name Otomyces represent this second stage of the Aspergilli also. This has been controverted by good authority, but the belief seems to be general that the Otomyces forms are connected with Aspergillus. Under the name Aspergillus, besides the forms whose life- histories are well worked out, which are known in both stages, mycologists include also a large number of forms known only in the first or asexual condition. Many of these may go on indefinitely in this stage and never develop further. Others possibly are but ill-understood variations of the better known forms. It is to this category of “Imperfect Fungi” that one or two of the more widely known species and all the species peculiar to the ear enumerated above are to be referred. The Aspergilli are among the commonest and most wide-spread of saprophytes, but in addition seem to find themselves at home in diseased animal tissues. Thus, in addition to the long list of Aspergilli which infest the human ear, A. fumigatus has been observed in the human lung, A. niger in the lungs of birds, A. malignus and A. nidulans are otherwise pathogenic, and A. virens has been found upon tissues imperfectly pre- served in alcohol. Penicillium crustaceum, the ordinary blue mould of cheese, fruit, jelly, etc., is the commonest of fungi. It is closely re- lated to Aspergillus, producing a spore-fruit of the same sort, ADDITION TO PARASITES OF HUMAN EAR 85 and is most readily distinguished in that the chains of conidia (asexual spores) proceed from verticillately branched hyphae instead of from a terminal swelling. This species also has a weakness for diseased tissues, having been found parasitic in other parts of the human body. P. minimum is known only as an ear-fungus. The Aspergilli proper, which furnish the bulk of the species infesting the ear and, according to report, are the fungi usually met with in cases of otitis parasitica, fall into three groups. In the first, the chains of conidia proceed from mere roughenings of the terminal vesicle of the fertile hypha. In the second, they proceed from well developed but simple sterigmata. In the third group these sterigmata are branched. The first two are included in the genus Aspergillus, the third has been made a distinct genus under the name of Sterigmatocystis. But systematists whotake into accountthe fur- ther development of S. nigra, the type, and its spore-fruit con- dition, now concur in uniting the genus with Aspergillus, so that it is kept separate chiefly because of the imperfect forms that are described under the other name. Sterigmatocystis nigra, or Aspergillus niger, is a very common saprophyte, only less common than A. glaucus and Penicillium crustaceum, and like them thoroughly pantogenous. WS. antacustica, an ear-parasite described by Cramer in 1859, was referred to Aspergillus niger by Wilhelm in his monograph of Aspergillus, and afterwards by Winter and Siebenmann. E. Fischer in the Pflanzenfamilien places it there doubtfully. There is good reason to suspect that A. niger is more common in the ear than the reports would show, as at least one figure labeled A. nigricans has branching sterigmata and the general ap- pearance of A. niger. Moilisia auriculae, a cup-fungus discovered in Italy in 1871 in a case of otitis, is known from a careful drawing made at the time by Dr. Frigerio. The name Peziza auriculae, under which it was published by Garovoglio in 1872, seems to have been a nomen nudum. The drawing shows a well developed spore fruit with asci and paraphyses as in typical Pezizineae. The Imperfect Fungi reported give rise to many difficulties. Trichothecium roseum is one of the most common of sapro- phytes and is pantogenous. On the other hand, Stemphylium polymorphum, Graphium penicillioides, and Coremium bicolor are saprophytes of decaying wood and are by no means so 86 ROSCOE POUND common or widely distributed. Siebenmann believes the two first to be connected with the form described as Verticil- lium graphii, the forms referred to G. penicillioides being only compact growths of the conidiophores. It will not escape notice that Verticillium, Acrostalagmus, Stachylidium, Tri- chothecium, and Graphium, the genera of Imperfect Fungi under which ear-fungi are described, have the common char- acteristic of being conidial stages of Hypocreales. We may suspect, therefore, that some single nectrioid fungus may ultimately be found to account for most of these, though the pantogenous T'richothecium roseum scarcely needs to be ac- counted for. Coremium bicolor was not found in the ear but in cultures of mycelia taken from the ear, and its place on the list is doubtful. Alysidium rufescens was first noted as a growth on the lens in cataract. Some time ago, Dr. S. E. Cook of Lincoln submitted to me material of an ear-parasite, plainly one of the Aspergillaceae, which had much of the outward appearance of the common A. candidus. Examination revealed branching sterigmata, and I referred the form provisionally to Sterigmatocystis candida Sace. The latter species was discovered by Saccardo in Italy in 1876 growing upon decaying insect larvae, and has since been found in France growing upon the surface of citric acid. It is probably pantogenous, like the rest of the group. While the form found by Dr. Cook differs from S. candida in being somewhat smaller at all points, the shape of the sterigmata is so characteristic, and agrees so thor- oughly with Saccardo’s figure (Fungi Italici, t. 80) that in the absence of authentic material for comparison, notwith- standing Professor Underwood’s caution that American fungi identified by European names are a source of confusion and must be renamed, it seems best to refer this form to S. candida, noting the slight divergence in measurements. A brief description, figures and bibliography are added. Sterigmatocystis candida Sacce. In a human ear affected with otitis, Lincoln, Neb. (Dr. S. FE. Cook). Fertile hyphea hyaline or whitish, rather strict, 150 to 200x10 » ; vesicle globose, 30 to 35 » ; basidia clavate, 30x74 y, noticeably obtuse and flattened at the top, bearing three filiform sterigmata 10 to 15» long; conidia gobose, not ex- ceeding 2 pz. ADDITION TO PARASITES OF HUMAN BAR 87 PRINCIPAL WORKS CONSULTED CaTTANEO, A. AND Otiva, L. Dei Miceti trovati sul corpo umano. 1883. Cooxr, M. C. On Some Remarkable Moulds. Journ. Queckett Micr. Club, II; 2:138. 1885. SIEBENMANN, F. Schimmelmycosen des Ohres. 1889. Neue botanische und klinische Beitraege zur Otomykose (1888), translated in Archives of Otology, 18:230, 1889. WILHELM, K. A. Beitraege zur Kenntniss der Pilzgattung Aspergillus. 1877. SaccARDo, P. A. Fungi Italici, t. 80. 1877. Sylloge Fungorum, vol. 4, 1886; vol. 8, 1889. FIscHER, A. Phycomycetes in Rabenhorst’s Kryptogamenflora v. Deutschland, 2d Ed. 1892. ENGLER, A. AND PRANTI, K. Die Natuerlichen Pflanzenfamilien, Th. I, Abt. 1, 119 et seq., 156, 297 et seq. 1893-1897. BURNETT, C. H. System of Diseases of the Ear, Nose, and Throat, vol. 1, pp. 190-203. 1893. Roosa, D. B. St. J. Diseases of the Ear, 7th Ed. 1891. 88 ADDITION TO PARASITES OF HUMAN EAR- EXPLANATION OF PLATE Plate XVIII Sterigmatocystis candida Sace. (a.) Fertile hypha. (b.) Terminal vesicle with “basidia” attached. (c.) A single “basidium.” PLATE XVII THE MODERN CONCEPTION OF THE STRUCTURE AND CLASSIFICATION OF DESMIDS, WITH A REVISION OF THE TRIBES, AND A REARRANGEMENT OF THE NORTH AMERICAN GENERA By CHARLES E. BESSEY, Pu. D. WITH ONE PLATE The recent revision of the Green algae in Engler and Prantl’s “Pflanzenfamilien” by Professor Wille, the eminent Swedish algologist, brings together in compact form the results of the work of many investigators. Taking this ad- mirable monograph as a basis and bringing to my aid the monograph of the Bacillariales by Professor Schuett in the same publication, I have ventured to attempt to carry out Wille’s work somewhat nearer to what appears to me must be its logical conclusion. I should associate in one group (Conjugatae) the families Zygnemaceae (including Mesocarp- aceae of some authors), Desmidiaceae, and Bacillariaceae (all of holophytic species), and to this group I assign ordinal rank. Until quite recently I have associated with these the families Mucoraceae and Entomophthoraceae, composed of hysterophytic plants, in accordance with the theory that they are colorless, degenerate relatives of the holophytic families just named. However, further study of the problem has led to the conclusion that Mucoraceae and Entomophthoraceae have little affinity with the families of the Conjugatae, and that they are to be removed to that remarkable group of hysterophytic families (Saprolegniaceae, Cladochytriaceae, Ancylistaceae and Peronosporaceae) in the Siphoneae, which appears to have sprung from or near the Vaucheriaceae. With these relationships this paper is not directly concerned, and they may be passed without further discussion. The families of the Conjugatae (Zygnemaceae, Desmidi- aceae, and Bacillariaceae) are here regarded as consisting of 90 CHARLES E. BESSEY typically filamentous plants, as is well illustrated in the com- mon Conjugata (Spirogyra) of the pools. As shown in another paper* many diatoms are filamentous plants, and in those species in which the cells occur singly we may regard this con- dition as the result of the early solution of the filament. In the present paper it is assumed that the Desmids, also, are typically filamentous, or in other words, that they have been derived from filamentous forms, a structure which is still maintained in a considerable number of genera, and that the unicellular condition is derived from this structure by the early separation of the cells, or as expressed above, by the solution of the filament. This conception necessitates an arrangement of the genera somewhat different from that adopted by Wille, without, however, seriously disturbing their inter-relationships. It is not difficult to see that the family is easily separable into three quite well-marked groups of genera, which we may, perhaps, regard as tribes. Thus the filamentous forms may be brought together (as indeed was done by Hansgirg and De Toni a dozen years ago), and in like manner the unicellular forms may be easily separated into two tribes, (a) those with elongated cells, little if at all constricted, and (6) those with broad, deeply constricted cells. To the first of these three tribes I have given the name Drsmipinan, preferring this to EUDESMIDIBPAE, used by Hansgirg for the name of his equiv- alent sub-family. The second tribe I name ARTHRODIEAD (from the genus Arthrodia, heretofore known as Clostervum, but clearly antedated by Rafinesque’s name) while for the third the name COSMARIEAE. In accordance with the foregoing conclusions I have drawn up the technical diagnosis of the family in the following terms: FAMILY DESMIDIACEAE Cells bright green, in unbranched filaments, cylindrical, angled or flattened in cross section, and quadrangular, rounded, or lobed and often constricted in side view; or more commonly separating early into isolated individuals which are similarly shaped, or symmetrically lobed or branched in side view; cell wall composed of cellulose, commonly finely *The Modern Conception of the Structure and Classification of Diatoms, with a revision of the tribes and a rearrangement of the North American genera; in Transactions of the American Microscopical Society, Vol. X XI, p. 61. CLASSIFICATION OF DESMIDS 91 porous, and often covered with a gelatinous layer, and com- posed in most genera of two halves which adhere to each other at the middle of the cell, which is usually constricted; propagation by the transverse fission of each cell into two equal, but unsymmetrical daughter cells, which soon grow to be symmetrical; generation by the rupture of the outer walls of two contiguous cells, and the protrusion of a thin-walled tube from each, these fusing and uniting their contents into a resting spore (zygote) from which on germination one, two, four, or eight new cells are formed.—Minute freshwater plants, floating free in the water of quiet pools, or entangled with Sphagna, mosses and other aquatic plants. KeEY TO THE TRIBES. A. Cells in unbranched filaments, Tribe 1. Desmidieae. B. Cells solitary, I. Cells elongated; not at all, or but moderately con- stricted, Tribe 2. Arthrodieae. II. Cells broad, deeply constricted, Tribe 3. Cosmarieae. TRIBE 1. DESMIDIEAB Cells in unbranched filaments, from much elongated to shorter than broad, cylindrical to angular or flattened, and from not at all to deeply constricted; filaments naked or en- closed in a hyaline sheath. KerY TO THE GENERA. I. Filaments naked (without a sheath), a. Cells cylindrical, 1. Chromatophore single, axial, 1. Gonatozygon. 2. Chromatophores three, parietal, spiral, 2. Genicularia. b. Cells barrel-shaped, 3. Gymnozyga. c. Cells quadrangular, deeply constricted, 4. Phymatodocis. II. Filaments surrounded by a hyaline sheath, a. Cells not constricted, or very little, 1. Filaments cylindrical, 5. Hyalotheca. 2. Filaments 3- to 4-angular, 6. Desmidium. b. Cells deeply constricted, filaments flattened, 1. Cells unarmed, 7. Sphaerozosma. 2. Cells armed with several divergent horns, 8. Onychonema. 92 CHARLES E. BESSEY 1. Gonatozygon De Bary. Cells elongated-cylindrical, or truncate-fusiform, attached to one another in an unbranched filament, which has no sheath, not at all constricted in the middle; chromatophore one, axial, undulated.—Small desmids of few species, rarely seen. 2. Genicularia De Bary. Cells elongated-cylindrical, at- tached to one another in an unbranched filament, which has no sheath, not at all constricted in the middle; chromato- phores three, parietal, spiral, sometimes confluent or irreg- ular.—Small desmids of few species, rarely seen. 3. Gymnozga Ehrenberg. Cells oblong, barrel-shaped, each with two median hoop-like ridges, attached to one another in an unbranched filament, which has no sheath, not con- stricted in the middle; chromatophores of several axial plates with divergent wings.—Small desmids of few species, several of which are common in quiet waters. 4. Phymatodocis Nordstedt. Cells oblong, truncate, quad- rangular in transection, attached to one another in an un- branched filament, which has no sheath, deeply constricted in the middle; chromatophores not known.—Small desmids, rarely seen. 5. Hyalotheca Ehrenberg. Cells short-cylindrical, attached to one another in an unbranched filament, which is sur- rounded by an ample, colorless sheath, very slightly (obtusely) constricted in the middle; chromatophores of several axial plates with divergent wings.—Small desmids of few species, several of which are frequent in some portions of this country. 6. Desmidium Agardh. Cells oblong, truncate, triangular or quadrangular in cross-section, little or not at all constricted in the middle, attached to one another in an unbranched fila- ment, which is surrounded by a hyaline sheath; chromato- phores of three or four longitudinal plates lying in the angles of the filament.—Small desmids, common throughout the country. 7. Sphaerozosma Corda. Cells compressed, deeply con- stricted in the middle, unarmed, ends rounded or truncate, slightly attached to one another in a lobed, unbranched fila- ment, which is surrounded by a hyaline sheath; chromato- phores quadriradiate——Small desmids, some species of which are common in ponds and ditches. CLASSIFICATION OF DESMIDS 93 8. Onychonema Wallich. Cells compressed, deeply con- stricted, armed with divergent horns, ends rounded or trun- cate, slightly attached to one another in a lobed, unbranched filament, which is surrounded by a hyaline sheath; chromato- phores quadriradiate.——Small desmids, rarely seen. TRIBE 2. ARTHRODIPAB Cells solitary, elongated, cylindrical to fusiform; transec- tion circular, not at all to moderately constricted; cells sheathless. Key To THD GENERA. I. Cells not constricted, transection circular, a. Cells straight, 1. Chromatophores of one or more spiral bands, 9. Entospira. 2. Chromatophore a single axial plate, . 10. Mesotaenium. 3. Chromatophores of several axial plates, with diverg- ent wings, 11. Peniwn. b. Cells more or less falcate, or semi-lunate, 12. Arthrodia. II. Cells straight, moderately constricted, transection circular. a. Chromatophores axial, 1. Cells short-cylindrical or fusiform, ends rounded, emarginately incised, 13. Tetmemorus. 2. Cells long-cylindrical, much elongated, ends trun- cate or rounded or 3-lobed, 14. Docidium. b. Chromatophores axial, 15. Pleurotaenium. 9. Entospira Brebisson (Spirotaenia Brebisson).* Cells soli- tary, sometimes aggregated in a gelatinous matrix, straight, oblong-cylindrical or fusiform, not constricted in the middle; transection circular, ends rounded or acuminate; chromato- phores of one or more spiral parietal bands.—In pools, ponds, and in wet mosses. 10. Mesotaenium Naegeli. Cells solitary, sometimes aggre- gated in a gelatinous matrix, short-cylindrical, elliptical or ovate, not constricted in the middle; transection circular, *Of these two names by the same author, Entospira has the priority, having been proposed by him in 1847 in Kuetzing’s Tabulae Phycologicae, while Spirotaenia did not appear until 1848, in Ralf’s British Desmidieae. 94 CHARLES E. BESSEY ends rounded; chromatophore a single axial plate or ribbon, sometimes divided in the middle—In pools, on wet rocks, walls or damp ground. 11. Penium Brebisson. Ceils solitary, sometimes aggre- gated in a gelatinous matrix, straight, cylindrical, or fusi- form, not constricted in the middle; transection circular, ends rounded or somewhat truncate; chromatophores of sey- eral axial plates, with divergent wings.—Large desmids, 11 to 80 » in diameter, and 6 to 10 times as long, common in pools and springs. 12. Arthrodia Rafinesque, (Closteriwm Nitasch).* Cells soli- tary, more or less faleate or lunate, incurved (rarely nearly straight), fusiform or cylindraceous, not constricted in the middle; transection circular, ends acuminate; chromato- phores of several axial plates, with divergent wings.— Medium to large sized desmids, 3 to 110 in diameter, and from 5 to 20 times as long, common in pools and springs. 13. Tetmemorus Ralfs. Cells solitary, straight, cylindrical, or fusiform, moderately constricted in the middle; transec- tion circular, ends rounded, narrowly emarginately incised; chromatophores axial—Rather large desmids, common in ponds. 14. Docidium Brebisson. Cells solitary, straight, oblong— cylindrical, moderately constricted in the middle, usually long (6 to 30 times their diameter); transection circular, ends truncate, rounded, three-lobed and three-spined; chromato- phores axial, of two to four radiating bands.—Large or medium sized desmids, frequent in ponds. 15. Pleurotaenium Naegeli. Cells solitary, straight, cylin- drical, more or less constricted in the middle; transection cir- cular, ends truncate; chromatophores parietal—Large des- mids, some species of which are common in ponds. TRIBE 3. COSMARIBAE Cells solitary, broad, more or less flattened; transection rounded to angular, oblong and elliptical, deeply constricted, the half-cells from entire to many-lobed; cells sheathless. *Nitzsch’s name, Closterium, was first proposed in 1817, while Rafinesque’s name, Arthrodia, was used four years earlier, i. e. 1813; hence the latter having clear priority. must be used for these beautiful organisms. Ne eee Oe er CLASSIFICATION OF DESMIDS 95 KEY TO THE GENERA. I. Cells short-eylindrical or orbicular, transection rounded or oblong, half-cells not lobed, a. Unarmed, 1. Solitary, a. Chromatophores axial, radiating, 16. Cosmarium. b. Chromatophores parietal, longitudinally lamini- form, 17. Pleurotaeniopsis. 2. Joined in gelatinous, branching threads, 18. Cosmocladium. b. Each half-cell armed with a spine on each side, 19. Arthrodesmus. II. Cells orbicular, oblong or elliptical, transection fiat- tened or elliptical, half-cells lobed, a. Half-cells with few, usually rounded lobes, and broad sinuses, 20. Euastrum. b. Half-cells with many pointed lobes and narrow sin- uses, 21. Micrasterias. III. Cells oblong or orbicular, transection rounded or ob- long or angular, a. Armed with spines, chromatophores parietal, lamini- form, 22. Xanthidium. b. Smooth, verrucose or hairy, chromatophores axial, 23. Stawrastrum. 16. Cosmarium Corda. Cells solitary, short-cylindrical or orbicular, smooth, verrucose, or rarely spiny, deeply con- stricted in the middle; transection sub-oval or oblong, ends rounded or truncate, entire; chromatophores one or two in each half cell, axial, radiating—Mostly small desmids of many species, widely distributed and common in mossy ponds. 17. Pleurotaeniopsis Lundell. Cells solitary, short-cylin- drical or rounded, unarmed, deeply constricted in the middle; transection sub-oval or circular, ends rounded or truncate; chromatophores parietal, longitudinally laminiform.—Med- ium to large sized desmids, a few of which may be found in our quiet waters. 18. Cosmocladium Brebisson. Cells joined in gelatinous, dichotomously branching threads, elliptic-reniform, con- stricted in the middle; chromatophore one in each half-cell, central.Small desmids of few species, but one of which has been found (in spring water) in this country. 96 CHARLES E. BESSEY 19. Arthrodesmus Ehrenberg. Cells solitary, short-cylin- drical or orbicular, smooth, with a single spine on each side of each half-cell, deeply constricted in the middle; transection oblong or fusiform-elliptical, ends rounded or truncate, en- tire; chromatophores axial, laminated.—Small to very small desmids, not common. 20. Huastrum Ehrenberg. Cells solitary, oblong or ellipti- cal, with few rounded lobes and broad sinuses, smooth or verrucose, deeply constricted in the middle; transection ob- long or elliptical, ends rounded or truncate, usually emargin- ate or deeply incised; chromatophore one in each half-cell, axial, of longitudinally radiating threads.—Small desmids of many species, widely distributed and quite common. 21. Micrasterias Agardh. Cells solitary, orbicular, or ob- long-elliptical, deeply constricted in the middle, each half-cell with three to five radiating, pointed lobes, separated by (usually) narrow sinuses, the lobes sometimes again divided; transection fusiform, ends entire, sinuate or incised; chroma- tophores axial, laminated.—Large desmids, common in mossy ponds and lakes. 22. Xanthidiwm Ehrenberg. Cells solitary or geminately connected, orbicular, inflated, armed with spines, deeply con- stricted in the middle; transection rounded, oblong or angular, ends neither emarginate nor incised; chromatophores parietal, laminiform.—Medium to small sized desmids, apparently not common. 23. Staurastrum Meyen. Cells solitary, oblong or orbicular, smooth, verrucose or hairy, deeply constricted in the middle, each half-cell in transection 3-to-6 or more angular, the angles often prolonged into obtuse or acute horn-like processes, ends mostly rounded or truncate; chromatophores axial.—Small desmids of many species, widely distributed but not abundant. : 4 ‘ 1 7 (All ¢ TRIBE 16. 19. CLASSIFICATION OF DESMIDS 97 EXPLANATION OF PLATE ells are drawn with their corresponding axes parallel, and are magnified about two hundred diameters.) Plate XIX DESMIDIEAE. . Two cells of a Gonatozygon filament. . Portion of a filament of Genicularia. Filament of Gymonzyga, with eight cells. Three cells of a filament of Phymatodocis. . Portion of a filament (eleven cells) of Hyalotheca, enclosed in a thick sheath. . Ten cells of a filament of Desmidium, enclosed in a sheath. . Portion of a filament of Sphaerozosma, enclosed in a sheath; two cells at the right just divided. . Filament of eleven cells of Onychonema, enclosed in a sheath. ARTHRODIEAE, . A cell of Entospira, with spiral chromatophore. . Cell of Mesotaenium. . Cell of Peniwm. . Cell of Arthrodia. . Cell of Tetmemorus. . Cell of Docidiwm. . A little more than one-half of a cell of Plewrotaeniwm. COSMARIEAE. Cell of Cosmarium. Cell of Arthrodesmus. . Cell of Luastrum. . Cell of Micrasterias. . Cell of Xanthidium. . Cell of Staurastrun. =~] PLATE XIX . 7 i F di F j i f f — P 2 + + an a - -. + ~ a ‘. ' < ( PHOTO-SPECTOGRAPHY OF COLORED FLUIDS By MOSES C. WHITE, M. D., NEw HAVEN, CONN. WITH ONE PLATE Every person with a moderate knowledge of mathematical optics is more or less familiar with what are known as the Fraunhofer lines in the solar spectrum, and they know too that incandescent vapors of different metals give character- istic bright lines in the spectra of flames in which the various metallic and non-metallic substances are volatilized. It is not so generally known that many fiuids, both colored and color- less, give characteristic dark lines when light passed through them is transmitted through the spectroscope. It is thus that the purity of olive oil may be determined by the spectroscope, which shows dark bands quite different from those produced by any other oil, intercepting the light admitted to the spectroscope. The examination of blood stains in medico-legal cases calls for some color test to distinguish blood from other colored fluids. It is well known that carmine, madder, alka- net, cineraria, permanganate of potash, wine, and many other fluids often look so much like blood as to give much trouble to distinguish them. The peculiarity of the spectrum as modified by the presence of blood has been ably described by many writers, and yet our books give only diagrams. Chemists give attention to the spectra of gases and astrono- mers are spending hundreds of thousands of dollars photo- graphing the light of the stars, and still more, photographing the corona of the sun in total eclipse of that orbit, hoping to learn more of the quality and condition of the matter compos- ing that wonderful luminary. But the character of the lines shown in the spectra of fluids is so different from the lines shown by incandescent gases, that I have thought it might be both interesting and useful to show by photography the absorption bands of blood and other colored fluids. 100 MOSES C. WHITE In entering upon this work we are met at once by the great difficulty of photographing red light. The portrait photogra- pher knows too well the red photographs, as blood and blue and violet appear in the photograph as almost pure white. Most colored fluids give their characteristic absorption bands in or near the red end of the spectrum, and it is well known that the so-called orthochromatic plates used by the modern photographer, while highly sensitive to blue and yellow, are almost wholly insensitive to ruby red, as it is ruby light and that only which is allowed in the dark room while developing modern dry plates. Plates then that are sensitive to red must be developed mostly in total darkness, and that is what astronomers do in preparing photographs of the sun and stars. Pure olive oil (Gallipole) in a tube 2 to # inch in diameter presents a dark shadow, or cutting out of the blue and violet ray with a fine almost indistinct line in the green and a strong dark band in the red. Refined cotton-seed oil presents the same appearance as re- gards the blue and violet only, but the blue and the red are continuous. If cotton-seed oil is mixed with the olive oil, there is no change in the blue and violet rays, but an almost entire fading out of the delicate line in the green and considerable diminution of the dark band in the red. With 50 per cent of cotton-seed oil the loss in intensity is considerable. With 25 per cent the variation is discernible.* To photograph the spectrum of colored fluids, specially pre- pared plates are required. To sensitize the plates for the red end of the spectrum cyanine is used according to the fol- lowing formula which comes to us from Germany. These plates will not retain their sensitiveness to red more than four or five days. The cyanine plates require long exposure, five or ten times as long as the same plates if not bathed in cyanine; hence fast plates are selected for this work. Take a solution of cyanine (1 to 1000) in absolute alcohol, 6 ccm. Codeia solution in absolute alcohol (1 to 1000), 34 ccm. Aniline oil, chemically pure, 5 drops. Distilled water, 960 ccm. In this mixture bathe the plates two minutes. Then rinse the plates one minute in alcohol, 34 cem.; * Chemical News, quoted in Scientific American, August 28, 1880, No. 243, p. 3874; also Olive Oil, Scientific American Supplement, June 28, 1884. PHOTO-SPECTOGRAPHY OF COLORED FLUIDS 101 distilled water, 966 ccm. Wipe the back of the plates with filter paper and dry rapidly in the dark; using chloride of calcium to hasten the drying. If successfully prepared, these plates are sensitive to all the colors of the spectrum, from the ultra violet to the extreme red. Figure S (Pl. XX) shows the entire length of the spectrum photographed as white of nearly uniform intensity. The solar spectrum is shown with the Fraunhofer lines sharply defined. In contrast with the Fraunhofer lines we see the absorption bands of blood (Fig. B, Pl. XX) wider, less sharply defined, and increasing in breadth as the solution is more dense. The line nearest the red, close to the line D of the solar spectrum, is more dense and more sharply defined than the second line (in the orange) which is broader and paler. Then we see the whole of the space belonging to the blue and violet is cut off by one broad absorption band produced by the blood. Didymium sulphate (the nitrate gives the same bands), a solution in water with a slightly pinkish hue, gives two ab- sorption bands—the first placed close to the D line in the solar spectrum, which is seen in the photograph (Fig. D, Pl. XX) as a brown shadow close to the left or red side of the first didymium line. The second didymium line, narrower and sharper than the first, lies about two-thirds of the distance from D toward E of the solar spectrum. In the blue and violet portions of the spectra of blood and didymium are seen lines produced by the carbons of the electric arc light used in pro- ducing the photographs. Figure P (Pl. XX) shows a series of beautiful lines of varying size and intensity in the spectrum of a solution of permangan- ate of potash. This colored fluid produces three heavy dark absorption bands in the green and a medium band in the orange and a very fine line in the yellow. It will be seen at once that many colored fluids are thus readily distinguished by the spectroscope. L. MOLOSSEZE ON SPECTRUM OF PICROCARMINATE OF AMMONIA.* We have long known the double absorption bands which are given by suitable solutions of alkaline carminates. I learned also from Ranvier that the picrocarminate of ammonia shows these two bands. But I did not believe that the spectra of * Archives de Physiologie Normale et Pathologique, Paris, 1877; 23, IV, pp 40-43. Translated by M. C. White, February 29, 1884, 102 MOSES C. WHITE picrocarminate at different degrees of concentration had been sufficiently studied, or that the resemblances and differences between blood and picrocarminate in solutions had been suffi- ciently described. When one examines with the spectroscope solutions of picrocarminate sufficiently concentrated, the red and the orange rays are alone transmitted. All that part of the spectrum beyond the sodium line, D, is obscured. The violet end of the spectrum is especially dark. In more dilute solutions the part previously dark is cleared up for a little distance from the line D and permits the green rays to pass. As we add still more water to the solution the dark part is cleared up still further, permitting the bluish-green (greenish-blue) to pass. There thus results to the right of D two absorption bands, the first, or the one nearest to D, narrower, while the second is the wider of the two. Continuing to dilute the picrocarminate the two absorption bands diminish a little in extent and become paler, the first more than the second; at the same time the right or violet end of the spectrum clears up more and more. At last the absorption bands entirely disappear, the second band being the last to disappear. If we compare this series of spectra with those given by haemoglobin in similar conditions, one is struck by the great similarity in concentrated solutions either of haemoglobin or picrocarminate, the red rays only being transmitted. In dilute solutions we find to the right of the sodium line two absorption bands, the first narrow, the second wider, while the right or violet end of the spectrum remains more or less obscure. There are, however, a certain number of differences between haemoglobin and picrocarmin- ate. In the spectrum of picrocarminate the part which first clears up when the solution is diluted little by little corre- sponds to the clear space which separates the two absorption bands the one from the other; the clear space which we find to the right of the two bands only appears later. In other words, the first absorption band is first separated from the dark part; the second band is separated later. In the spectra of haemoglobin, on the contrary, the part which first clears up is that which is to the right of both bands of absorption, while the bands themselves are only separated when the solu- tion becomes more dilute. The absorption bands of picrocar- minate are a little more distant from the sodium line and a PHOTO-SPECTOGRAPHY OF COLORED FLUIDS 103 little more distant from each other than that of haemoglobin. The clear space which separates the bands of picrocarminate corresponds to the second band of haemoglobin. When we continue to dilute, the solutions of picrocarminate grow pale more rapidly and it is the first band (the one nearest to the sodium line) which disappears first. The bands of haemoglo- bin remain longer, clear and distinct and the second band is the first to disappear. If one does not fully understand the phenomena which solutions of picrocarminate present he might say that the spectra of haemoglobin and picrocarminate are but little different from each other. The spectrum of picro- carminate is explained by that of carmine and by that of picric acid. The two absorption bands of picrocarminate ap- pear and disappear in the same way as those of carmine. Their positions and intensities are the same. But in the spectrum of carmine the extreme right becomes clear rapidly when the solution is diluted, while it remains dark much longer in the spectrum of picrocarminate. This obscurity is due to the presence of picric acid. The spectrum of this coloring matter is dark at the right (at the violet end) and becomes clear very slowly. The spectrum of picrocarminate is thus a true combination between the spec- trum of carmine and that of picric acid. Note.—At the New York meeting, when Dr. White read this paper, he was, despite advancing years, one of the most active and interested members present, participating in all the discussions with keen enjoyment. He did not have an opportunity to correct the proofs of this article before his last illness had come upon him. Con- sequently the reader will find no doubt that there are minor errors which have escaped the editor’s notice, but which would have been rectified by the author. 104 PHOTO-SPECTOGRAPHY OF COLORED FLUIDS EXPLANATION OF PLATE All figures after photographs by M. C. White. Plate XX Spectrum of Blood. Spectrum of Didymium Sulphate. Spectrum of Picrocarminate. Spectrum of Permanganate of Potash. Solar Spectrum with Fraunhofer Lines. myoouw PLATEK XX DESCRIPTION OF A NEW GENUS OF NORTH AMERI- CAN WATER MITES, WITH OBSERVATIONS ON THE CLASSIFICATION OF THE GROUP By ROBERT H. WOLCOTT WITH ONE PLATE T. STHGANAPSIS nov. gen. Diagnosis of the genus: An hydrachnid of the family Hygrobatidae, characterized by ithe possession of a very delicate exoskeleton in the form of a continuous open mesh- work; with palpi of the character of those of Arrenurus, the four basal segments being stout, the fifth small and opposable to the distal end of the fourth; with epimera resembling in form and arrangement those of Arrenurus; with legs bearing swimming hairs; and with the genital opening guarded by valves and these in turn flanked by numerous acetabula im- bedded in the wall of the body. At first glance, and as seen from the ventral surface the species which is the type of this genus was taken for an Arrenurus on account of the form and relationships of the epimera, and the general character of the structures about the genital opening. This impression was strengthened by an examination of the palpi. But when turned over, the absence of the characteristic chitinous exoskeleton of Arren- urus with its prominent furrow, and the general appearance of the specimen as seen from the other side dispelled the illusion. A careful examination shows that it differs in many and important details from the species included in Arrenurus, and that it is evidently a type of a related genus, hitherto undescribed, and intermediate between the genus named and some soft-bodied form, from which perhaps that genus is descended. The name is derived from the Greek steganos, covered, and apsis, a mesh, and is bestowed in allusion to the chitinous 106 ROBERT H. WOLCOTT exoskeleton which gives to the mite, when mounted on a slide and viewed as a transparent object, a characteristic appearance. Steganapsis arrenuroides noy. sp. The body of the single specimen in the author’s collection is of medium size and broadly elliptical in form, the width being almost precisely three-fourths of the length. It is evenly rounded at both ends, strongly convex above, moder- ately convex below, and flattened over the epimeral area. The hairs which it bears are long and very slender; on either side of the posterior end of the body is an extremely long and fine hair, and at its outer side a shorter one also very fine. The cuticula is marked by rather conspicuous irregular striae, and beneath it is an exoskeleton consisting of a con- tinuous meshwork (Pl. —, Fig. 1), the openings in which are, over the body, large, practically uniform in size, and with irregular rounded outline. The epimera, the exoskeleton of the palpi and that of the legs are made up of a similar meshwork with finer meshes and with proportionately heavier trabeculae, and this diminution in size of mesh and in- crease in coarseness of trabeculae is, in any particular part, greater or less, apparently in direct proportion to the strength desirable. At the extreme ends of the epimera about the insertion of the legs the chitin is very thick, and without any trace of such meshes. This meshwork on surface view pre- sents an appearance in a certain degree resembling that seen in Arrenurus, but the meshes contain no granular, gland-like cells such as those which occupy the goblet-shaped pores in the thick exoskeleton of that genus and of Arendowskia. The eyes are large, widely separated, and close to the lateral margins of the body. The maxillary shield is broad and relatively short, with parallel sides and evenly rounded posterior margin. The rostrum (Pl. XXI, Fig. 2, r.), rather similar to that of certain species of Arrenurus, is short and blunt. The mandibles (Pl. XXI, Fig. 3) are more slender than in any species of Arrenurus examined, and the proximal segment tapers towards its distal end, becoming, at the articulation with the claw, of the same breadth as the claw which itself is slender and strongly curved. The palpi (Pl. XXI, Fig. 2) are short and stout with a rela- NEW GENUS OF NORTH AMERICAN WATER MITES 107 tively short ventral margin, and long dorsal margin as seen in Arrenurus. Segment 1 is small and narrow; 2, 3, and 4 of about equal thickness, and 5 a mere claw. Segment 2 has a very convex dorsal margin and a shorter straight ventral margin. It possesses several spines, one in the middle of the dorsal margin, two short ones on the distal portion of the median surface toward the dorsal side, and three longer toward the ventral side. Segment 3 is nearly triangular with a very short ventral margin; it bears a long slender spine on the inner surface nearer the dorsal than the ventral mar- gin. The dorsal margin of 4 is quite convex, the ventral moderately so, the distal end being a little over one-third the diameter of the proximal and bearing the claw-like distal segment. This segment 4 bears a flat keel-like expansion which begins on the ventral surface about one-third the way from the proximal end and passes around the distal end to the dorsal surface, its margin forming a rounded angle a little less than a right angle. This keel projects beyond the base of segment 5 at its outer side and bears a coarse spine on its inner surface. The epimera (P]. XX1, Fig. 4) form three masses, I and II on either side being approximated, and the first pair being fused along their median edges, the suture between them becoming quite obliterated posteriorly; the median posterior margin of this pair is also indistinct. The space between II and III is of moderate width and less than that separating III and IV of opposite sides. The anterior and median border of this mass forms an even curve, the posterior is straight and runs slightly backward forming a blunt angle towards the outer side. The external ends of the epimera are mod- erately produced and rounded. The legs are of moderate length, only the last exceeding the length of the body and that by very little. Leg I is about two-thirds the length of IV and II and III intermediate, with III a trifle longer than II. The segments are, in II and III, in order of length 6, 4, 5, 3, 2, 1; in I, 5 is equal to 4 or even a little longer, and in IV, 4 is even longer than 6. The legs increase regularly in stoutness from I to IV, and the segments from the tip of each leg to the base. The distal segment of each leg bears a pair of small and extremely slender, sharply pointed, simple claws. Spines and hairs are numerous on the legs, and many of them extreme in their 108 ROBERT H. WOLCOTT length and fineness. Legs I and II possess a clump of long slender spines at the tip of segment 2, and another at the base of 3, and two long and stouter ones at the tips of 3, 4, and 5, while other spines are few and stout. Leg III has similar spines on 2 and 3, and numerous long hairs toward the tips of 3, 4, and 5, fewer in number on 5. On IV are a great many spines at the tip of 2 and along the whole ventral surface of 8, 4, and 5, varying in length and coarse- ness, and a profusion of swimming hairs of extreme length and fineness. The distal ends of all segments but the basal of each leg are produced on the anterior and posterior sides, beyond the insertion of the next segment, into two processes; these on segments 2 and 3 of each leg are sharply pointed, while on 4 of each a spine is inserted into the extensor side of this process and on 5, this spine being at the tip, the pro- cess becomes bifid. This process varies on different segments but in general the anterior is more prominent than the pos- terior on any particular segment, and the processes on leg IV are less prominent than on the anterior legs. (See Pl. X XI, Fig. 5.) The genital area (Pl. XXI, Fig. 4) of the specimen under ob- servation, which is apparently a female, resembles very closely that of a female Arrenurus. The opening is flanked by two movable lunate valves, and beyond these numerous acetabula, occurring in an area which is in the form of a band somewhat less in width than the length of these valves, and which runs backward, upward and outward for a short dis- tance on the sides of the body where its end is evenly rounded. These acetabula are set into the meshes in the chitinous exo- skeleton. Between this tract and the posterior epimeron of either side is a gland, and posterior to the genital opening and midway between it and the end of the body is the anus. Measurements: Length of body, exclusive of claw....0.868 mm. Total length of palpi (average of two).0.204 mm. Length of leg I, exclusive of claw....0.617 mm. Length of leg II, exclusive of claw. ..0.699 mm. Length of leg III, exclusive of claw. .0.729 mm. Length of leg IV, exclusive of claw. .0.908 mm. For the single specimen in the collection of the author he is indebted to Mr. James B. Shearer, of Bay City, Michigan, NEW GENUS OF NORTH AMERICAN WATER MITES 109 who collected it at Les Chenaux Islands in northern Lake Huron, during August, 1895. The name given is suggested by the very close superficial resemblance between this form and the females of Arrenurus. II. KRENDOWSKIA OVATA Wolcott Wolcott, 1900: 181, Pl. IX, Figs. 1-7. In material collected in Lamberton Lake near Grand Rapids, Michigan, July 22, 1898, were found six specimens of this species including four nymphs. Both the adults are males with three acetabula on either valve, and in neither is the proboscis protruded. The nymphs (Pl. XXI, Fig. 6) are about 0.4 mm. in length, and present the broadly oval outline and the strongly convex dorsal surface of the body characteristic of the adult. The surface of the body is covered by an exoskeleton in the form of a delicate chitinous meshwork with fine meshes. There are numerous long hairs, especially long at the posterior end of the body, and a pair just within and anterior to each eye. The palpi are also similar in form to those of the adult. The epimera have much the same form and relationships except that the first two are fused in the median line and the space included between the ends of all seems to be filled in with a thin sheet of chitin. The last pair do not show the character- istic excavation toward the median end. The legs are spar- ingly furnished with relatively coarse spines and at the distal end of segments III, 5 and IV, 5 and IV, 6 are a few very long and very fine swimming hairs. The genital area is occupied by a chitinous plate which has in general the form of an equilateral triangle all the angles of which are truncated and rounded, and which lies with a more strongly truncated apex anteriorly and with an emarginate base posteriorly. It bears two acetabula on either side. Thor, in a private communication, has expressed the view that this species is identical with his Geayia venezuelae, in which case the name should stand Krendowskia venezuelae (Thor). III. Komnrkna concava Wolcott Wolcott, 1900: 190, Pl. XI, Figs. 15-22, and Pl. XII, Figs. 23, 24. The result of recent examination of material previously unassorted, and re-examination of material collected several 110 ROBERT H. WOLCOTT years ago, has been to extend the known range of this species over a wide area. Specimens are in the author’s collection from the following localities: Massachusetts: Cranberry Lake, Wood’s Hole, July 28 and August 138, 1900. Michigan: Lake St. Clair, August and September, 1893. Susan, “26,” and Twin Lakes, Charlevoix, August 6 and 21, 1894. Reed’s, Fisk’s, Round, Soft-water, Lamberton, Muir’s, Dean’s and Power’s Lakes, and Grand River, Grand Rapids, June 27, 1895, and at various dates during July and August, 1895, 96, ’97, ’98. Illinois: Fox Lake, Lake County. September 17, 1894 (coll. by H. B. Ward). Nebraska: Pond at State Fish Hatchery, South Bend, Sep- tember 2, 1897. Pond at Wayne, September 8 and 9, 1899 (coll. by Miss Caroline Stringer). It is a pleasure to record the fact that this genus, dedicated to the author of the first paper of real value on North Ameri- can hydrachnids, is not only a most attractive form but also by virtue of its wide distribution a characteristic member of our hydrachnid fauna. IV. TANOAGNATHUS SPINIPES Wolcott Wolcott, 1900: 194, Pl. XIJ, Figs. 25-28. A second specimen of this species, collected in Lamberton Lake near Grand Rapids, Michigan, July 22, 1898, belongs to the same sex as the type specimen and agrees with it in every respect. YV. OBSERVATIONS ON THE CLASSIFICATION OF WATER MITES Among the recent additions to the literature of the Water Mites, the papers of Thor are noteworthy. Since the work of Neuman, which culminated in his “Sveriges Hydrachnider,” for many years the most pretentious work on the subject, no Scandinavian student of the group had appeared, until in 1897 Thor began a series of papers which has become both numer- ous and valuable, and has given us a very complete knowledge NEW GENUS OF NORTH AMERICAN WATER MITES LiL of the Norwegian hydrachnids. Among the most recent of these is a “Prodromus” (Thor, ’00) in which he suggests a division of the group into fourteen families: Limnocharidae, Eylaidae, Hydryphantidae, Hydrachnidae, Lebertiidae, Sper- chonidae, Limnesiidae, Hygrobatidae, Pionidae, Curvipedidae, Atacidae, Brachypodidae, Aturidae and Arrenuridae. Al- though these families are not characterized and the basis on which they rest is not fully apparent, the paper is interest- ing to a student of the group as being the first attempt to break up the Hygrobatidae—or Hygrobatinae, as one pleases —ijinto smaller groups, an attempt induced by the rapid in- crease in the number of described genera. It seems to the writer, however, that in certain respects the proposed arrange- ment is decidedly a step backward since the families suggested do not represent groups of equivalent value, while its author seems to have passed over all that had been previously ac- complished. Even in the writings of very early writers is observable a tendency to partition the genera of Water Mites, then few in number, between groups of higher rank, and Leach (15) in 1815 recognized two families; the Eylaides with one genus, Eylais, and the Hydrachnides with two genera, Hydrachna and Limnochares. Koch (87), in 1842, included under the order Acari, Section 1 or Water Mites, containing forms with movable swimming hairs, two families, Hygrobatides and Hydrachnides. To the former, characterized by the possession of two eyes, he assigned the genera Ataxz, Nesaea, Piona, Hygrobates, Hydrochoreutes, Arrenurus, Atractides, Acercus, Diplodontus and Marica; to the latter, characterized by the presence of four eyes, Limnesia, Hydrachna, Hydryphantes, Hydrodroma and Eylais. Under Section 2, or Swamp Mites, containing forms with. a prominent snout, he included Limnochares, Thyas, Smaris and Alycus, of which neither of the last two have been again grouped with the Water Mites until Nordenskiold’s proposed arrangement, given hereafter. Kramer (77) divided the hydrachnids into four families: Hydrachnidae, with mandibles one-jointed, lancet-shaped (type Hydrachna); Eylaidae, with mandibles dwarfed and con- sisting of two hooklets (type Hylais); Hygrobatidae, with mandibles distinctly two-jointed, the distal claw-like (types Nesaea and Arrenurus); and Limnocharidae, with mandibles and labrum fused forming a firm capitulum (type Limnochares). 112 ROBERT H. WOLCOTT Haller (81a) gave to the “Hydrachnidae” the rank of a sub- order and recognized two families on the basis of the position of the eyes; Medioculatae, including Limnochares and Hylais, and Lateroculatae, including all the other genera. In 1891. Canestrini (91) raised the “Hydracarini” to the rank of an order, and included under it three families—Halacaridae, Limnocharidae and Hydrachnidae. The first, or Marine Mites, are by most authorities separated entirely from the Water Mites, while of the two other families the former is equivalent to Haller’s Medioculatae, and the latter to his Lateroculatae. Thus the matter of systematic arrangement stood, when in a later paper Kramer (93) by distinguishing clearly between three principal types of larvae, and making this the basis of his arrangement of the genera, proposed the following, which seems to the writer the first logical classification: Order Prostigmata— Fam. 1—Hydrachnidae (larval type I). Fam. 2—Hygrobatidae (larval type II). Fam. 3—Eylaidae (larval type III). Fam. 4—Trombididae. Piersig (97) in his great work on “Deutschlands Hydrach- niden” reduced the Hydrachnidae to the rank of a family with five sub-families—Limnocharinae, Hydrachninae, Ey- lainae, Hydryphantinae, and Hygrobatinae. Nordenskiold (98) later canvassed very carefully the morphological and sys- tematic relationships of the Hydrachnidae and came to the conclusion that the term Trombididae should include a num- ber of sub-families, representing two lines of descent, to the forms included under one of which he applied the term Trom- bidiformes, and to the others Rhyncholophiformes. The for- mer contains, according to him, the sub-families Trombidinae, Hydryphantinae, and Hygrobatinae on the one hand, and Eylainae and Limnocharinae on the other; the latter the Hydrachninae, Rhyncholophinae, including the genus Rhyn- cholophus, etc., and Smaridinae, including Smaris, Smaridia, ete, From an examination of the literature accessible the opin- ion seems to be quite generally held among recent authors that the Water Mites as a whole form a group of higher rank NEW GENUS OF NORTH AMERICAN WATER MITES eles than that of a family, and in the judgment of the writer this opinion is well founded. A careful consideration of the sub- ject involving an earnest study of previously proposed ar- rangements and a careful weighing of the value of the char- acters offered by these forms has so far not led to results that the author is willing to publish as final, but the following tentative scheme is suggested to indicate the relationships of the group and its components: Phylum—Arthropoda, Class—A rachnoidea, Order—Acarina, Sub-order—Prostigmaia, Tribe I—Trombidini, Tribe [1—Hydracarini, Fam. 1—Hydrachnidae, Fam. 2—Limnocharidae, Sub-fam. 1—Limnocharinae, Sub-fam. 2—Hylainae, Sub-fam. 3—Hydryphantinae, Fam. 3—Hygrobatidae, Sub-families? It seems to the writer that no argument is necessary to ex- plain the use of the terms above down to that of sub-order. He believes also that Kramer’s position in regard to the natural character of the group Prostigmata is well taken, but it does not seem that the characters of this group are such as to be accorded more than sub-ordinal rank. The separation of this sub-order into the tribes Trombidini and Hydracarini is based on views suggested or expressed by sev- eral authors. The former is practically synonymous with Canestrini’s Trombidina plus his Hoplopina, which do not seem to be co-ordinate groups, while the Halacaridae are thrown out of the Hydracarini, since as Trouessart (89) says, they seem to be intermediate between the Gamasidae and Sarcoptidae. As thus limited the Hydracarini form a well- defined and very homogeneous group, certain forms in which furnish an easy transition to the Trombidini. 8 114 ROBHPRT H. WOLCOTT The three families suggested are based on Kramer’s ar- rangement, and are separated not only by characteristic larval types but also by structural characters of the adult, especially such as relate to the form and relationships of the mouth- parts. The sub-families of the Limnocharidae are also based on both larval and adult characters. As to the possibility of further division of the Hygrobatidae on characters that are essential or natural the author at this time reserves his opin- ion. He would, however, present for consideration the fol- lowing propositions: 1. The tribe Hydracarina as defined above is polyphyletic in origin as evidenced by the wide difference between the three larval types. 2. The uniformity of conditions under which the species exist has led to a great similarity of structure among adults. 3. The group becomes thus both sharply limited and very homogeneous. 4. The characters of the early stages reveal more of the phylogenetic relationship than do those of the adult. 5. On the other hand, owing to the same uniformity of con- ditions the forms are unusually conservative, and specimens from widely separated regions and very different localities show practically no variation. 6. The structural characters presented by the adult may in consequence be accepted as possessing a maximum value in classification. 8. In the division of the tribe into families and then into sub-families the natural grouping of the genera must depend on the characteristics of the larvae, to which should be added in the definition of the group the common structural charac- ters of the adult. Certain facts brought out in the study referred to earlier in this paper support these views. The genus Koenikea illus- trates very well the constancy of the forms. Specimens col- lected in a small shallow pond (Cranberry Lake, Wood’s Hole, Mass.) nowhere over two feet deep, and covering an area of but an acre or two, and lying out on a peninsula with a bluff on the one hand and on the other the Atlantic Ocean, from which it is separated by a narrow strip of sand; others from a lake (Lake St. Clair, Mich.) covering over 400 square miles and with a very uniform depth over the center of about 20 feet; others from lakes (near Grand Rapids, Mich.) of half NEW GENUS OF NORTH AMERICAN WATER MITES 115 a mile to a mile and a half in diameter, and 60 to 80 feet in depth; others from a small artificial pond (South Bend, Nebr.) separated by 1,500 miles from the first locality named; others from a narrow prairie slough (Wayne, Nebr.) the rem- nants of a former creek bed; and still others from among the reeds and rushes in a rapidly flowing river (Grand River, Mich.)—all are on careful examination quite indistinguish- able, and altogether nearly 100 specimens have been examined without the detection of an appreciable variation. The homogeneous character of the group must be evident to every student of these forms, at least if his experience be similar to that of the writer, who approached the study of them quite unprepared for the wonderful variety exhibited by the details of structure which have become apparent after careful examination, where at first on superficial examination all looked practically alike. The temptation is strong to continue these observations on the classification of the forms with which we are dealing, but further discussion concerning the more comprehensive groups and a consideration of the basis upon which the genera in the group should rest is withheld, to be published in another paper. Zoological Laboratory, The University of Nebraska, 116 NEW GENUS OF NORTH AMERICAN WATER MITES LITERATURE CANESTRINI, GIOVANNI. 91. Abbozzo del Sistema Acarologico. Estr. dagli. Atti del R. Istituto veneto di scienze, lettere ed arti; Tomo II, Ser. VII, 699-725. Venezia, 1891. HALLER, G. 8ia. Die Hydrachniden der Schweiz. Mittheil. der Bern. Naturf. Ges.; Jhg. 1881, 2 Hft., 18-83, Taf. I-IV. Published separately; Bern, 1882. Koca, C. UL. 37. Uebersicht des Arachnidensystems, 3 Hft., 7-39, Tab. I-IV. Nurnberg, 1842. KRAMER, P. Tite Grundzuege zur Systematik der Milben. Arch. f. Natur- gesch.; XLIII Jhg., Bd. I, 215-247. 93. Ueber die verschiedenen Typen der sechsfuessigen Larven bei den Suesswassermilben. Arch. f. Naturgesch.; LIX Jhg., Bd. I, 1-24, Taf. I. Lracn, W. E. 15. A tabular view of the external characters of four classes of Animals, which Linné arranged under Insecta, ete. Trans. Linn. Soe. London; XI, 306-400. NORDENSKIOLD, ERIK. 98. Beitrage zur Kenntnis der Morphologie und Systematik der Hydrachniden. Acta Soc. Scient. Fennicae; XXIV (1898), 1-75, Pls. I, II. Helsingfors, 1898. PIERSIG, R. 97. Deutschlands Hydrachniden. Bibliotheca Zoologica, XXII. ViII+601 pp., Pls. I-LI. Stuttgart, 1879-1900. THOR, SIG. 00. Prodromus Systematis Hydrachidarum. Nyt. Magazin for Naturvidenskab; XXXVIII, Hft. 8, 1-4. Kristiania, Sept., 1900. Second edition: ibid., 263-266; Nov., 1900. TROUESSART, E. L. 89. Revue Synoptique de la Famille des Halacaridae. Bull. Scient. de la France et de la Belg.; XX, 225-251. WotcorttT, R. H. 00. New Genera and Species of North American Hydrachnidae. Studies from the Zool. Lab. Univ. of Nebr., No. 34. Trans. Am. Micros. Soc.; XXI (1899), 177-200, Pls. IX-XII. (June, 1900). NEW GENUS OF NORTH AMERICAN WATER MITES 117 EXPLANATION OF PLATE All figures drawn with the aid of a camera lucida—but in Figs. 1 and 3 only certain dimensions thus secured, the rest sketched by eye. Plate XXI All but Fig. 6, of Steganapsis arrenuroides nov. sp. Fig. 1. A portion of the meshwork formed by the exoskeleton, from near the base of one of the palpi and showing the opening of a gland. X about 375. Fig. 2. The palpi and rostrum (r.). X 250. Fig. 3. The right mandible. X 250. Fig. 4. The epimeral and genital areas, drawn from the pre- served specimen before being dissected and the parts mounted on a slide. X 60. Fig. 5. Anterior side of distal segments of the left fore leg. xX 192. Fig. 6. Ventral view of the nymph of Krendowskia ovata Wole. xX 75. XXII PLATE THE CLADOCERA OF NEBRASKA* By CHARLES FORDYCE, NEBRASKA WESLEYAN UNIVERSITY WITH FOUR PLATES INTRODUCTION In the fall of 1895 the writer began a study of the Cladocera of Nebraska; the work on this group has continued with in- creasing interest for a period of five years, with such results as may be found in this paper, which has been prepared in connection with a course of graduate study in the Depart- ment of Zoology in the University of Nebraska and accepted by the Faculty, June, 1900, as a thesis for the degree of Doctor of Philosophy. Gratitude is due Dr. R. H. Wolcott and Miss Caroline E. Stringer for several bottles of material received from them, and the writer would express his most grateful acknowledg- ments to his friend Dr. H. B. Ward, of the University of Nebraska; for it was through his influence and kind personal guidance that the task has been pursued and finally com- pleted. METHODS The material was obtained partly with the Birge net and partly by the use of a small plankton pump which was de- vised by the writer during the early part of his work, Fordyce (98). It has proved to be a very valuable piece of collecting apparatus as it enables one to collect in many places inacces- sible to the ordinary net, and by means of it both the qualita- tive and quantitative value of the collections may be ascer- tained as well as the vertical distribution of the fauna. The material is preserved partly in formalin, partly in alcohol and partly in a solution consisting of: GIy GSriMe wy qasciece cies stikerereiee sle, cis: oicie 2 parts by vol. Distilledsp waters sscic- c-) et ge Pe «2? Ur eee id oe at UF Bea teks ae, Seg oe Fie. 1 PLATE XXVIII tse REM a Ste aaa. at _gungene” REPORT OF THE LIMNOLOGICAL COMMISSION The initial report of a body so recently organized as this can hardly be more than preliminary in character, all the more so that the field entrusted to it is as extensive as untried. When by the action of this Society a year ago the Limnological Com- mission was organized and its members asked to assume the duties laid upon them in connection therewith, they accepted not without some hesitancy at the extent of the work before them. The study of fresh water bodies is indeed a great field, barely touched upon at one or two points in this country and nowhere in the world even superficially covered as yet. Nev- ertheless it was the original field of biologic study; it was and is accessible to public and private workers practically every- where and affords opportunities for extended or limited work in any particular department of biologic research towards which the student may be drawn. Furthermore, to this work attaches an undoubted interest for all who come within its territory, while its problems have not only great biologic im- portance but are also of economic value as well as of decidedly practical character, touching as they do upon the important questions connected with fish culture, municipal water supply and sewage disposal. In this first report it will not be possible to do more than outline succinctly what has developed from our correspond- ence and discussion thus far regarding the object of the work, to make a brief survey of the field under discussion, of the ends to be reached and of some of the means for attaining them, and finally to invite propositions concerning the meth- ods and probiems under consideration and co-operation in pro- ceeding towards their solution. It may be fitting at the outset to state briefly the outlook before the Commission. Such a venture as this is not entirely unheard of, and consequently venturesome. A similar body was appointed some years ago by the Swiss National Society for Natural Sciences. As a Swiss investigator, Professor Forel, of Geneva, was the pioneer in the study of fresh water lakes, and as the investigators of this beautiful mountain re- 13 194 REPORT OF LIMNOLOGICAL COMMISSION public have retained their supremacy in this field of research through more than thirty years, so also Switzerland was the leader in organized effort towards the development of lim- nological investigation. The plan of the Swiss Limnological Commission in assigning work in various regions to different students has met with such success as to inspire those who follow in its footsteps with hope for the outcome of their efforts and as to hold up a high standard for their attainment. Similar results can not be expected in a brief period of time, but we hope that they may be reached here eventually. The study of limnologic questions affords abundant oppor- tunities for workers of every type and of every grade; but if the results of such varied activity are to be of permanent value or of general import, they must be correllated and unified. Therewith gaps in the record will become apparent and new problems be suggested so that the lines of work will be ex- tended and at the same time joined together into a symmetri- cal system. The fundamental objects then of this Limnologi- cal Commission we believe to be: To co-ordinate the results obtained by different investigat- ors into a united whole; to enlist new workers and to en- courage new work along lines already marked out; to suggest new lines of work and methods of research, and to aim at uniformity of procedure so that the results may be compared and correllated. For convenience in discussion and in the organization of the work, the field of limnologic study has been cut up into a number of main’ divisions and some of the chief subdivis- ions under each indicated. These are as follows: 1. Bibliography: a general historical review of limnologi- cal studies to date; periodic summaries of work done in the world at intervals thereafter. 2. Physiography: the inanimate environment, including the physical and chemical study of water bodies; types of such bodies, distribution; temperature, color, circulation; lake areas; composition of water, etc. 3. Biology, (a) Taxonomy of water organisms: systematic tables, description and sketch of each on cards to form eventu- ally a faunal catalogue for the United States. (b) Morphology of organisms: anatomy, histology, embryology of individual forms. (c) Distribution of organisms: (1) Geographie; (2) regional: littoral, limnetic, bathybic species; (3) quantitative: REPORT OF LIMNOLOGICAL COMMISSION 195 general, numerical, proportional. (d) Physiology, experi- mental studies. (e) Ecology. 4, Applied limnology: water supply, sewage, fish culture. After this preliminary statement, the Limnological Com- mission has the following recommendations to make for the purpose of advancing this work: First, it is expedient that, as soon as suitable persons can be found who are willing to undertake the work, there should be added to the Commission a physicist, a chemist and a bac- teriologist, in order that these phases of the environment may be adequately studied. Second, the influence of the Society should be directed to- wards the production and publication of accurate systematic accounts of the fresh water organisms, to the end that the various workers on limnologic questions may have at hand taxonomic summaries of the organisms with which they come in contact. It is not too much to say that such treatises are non-existent for American forms and inaccessible to the ma- jority even for the few groups which have been partially worked out. This must be the first step in the inauguration of the proposed movement. The publication of a series of cata- logue cards, each devoted to a single species, appears as a desirable method of putting such data into accessible form ~ and keeping them in shape for frequent emendation or ad- dition. Third, in the interest of a complete knowledge of the dis- tribution of fresh water organisms, the Commission plans the keeping of careful faunal records. It is proposed to appoint one or two investigators for each group who shall undertake to enter and collate all faunal records of this group which may be sent them and conversely to furnish workers with in- formation concerning the distribution of such organisms. This plan will ultimately yield data for the discussion of the geographical distribution of fresh water genera and species. It will also enable the elimination of such data as are common, leaving for publication by the student those facts which are important for one reason or another. Fourth, the Commission is of the opinion that an occasional summary of progress in the field of Limnology will serve to keep students in touch with the subject by giving them knowl- edge of the work of the world in general. This is that sub- division of the field which stands first in the outline given 196 REPORT OF LIMNOLOGICAL COMMISSION above. It has been covered sufficiently for the present by the summary and review printed in the Transactions of this So- ciety, volume XX, bringing the subject up to January, 1899. Fifth, the Commission would most strongly advise that in- dividual work should be limited to a single body of water or to a definite problem studied with reference to a series of such water bodies. The results will be most useful for all pur- poses when they bear upon the thorough treatment of a single phase of the subject rather than more indefinitely upon a wider field. There is naturally involved in the effort to carry out such plans as have been outlined some expenditure of money, even if the services of various investigators are freely and gra- tuitously placed at the disposal of the Commission. Accord- ingly, an appeal is made herewith to the generosity of those interested in the movement and in the development of bio- logical study in our country for contributions, large or small, for the prosecution of this work. In conclusion, all students interested in this subject are in- vited to participate in the work. It is by general and gen- erous co-operation that success will be attained. The student who is working alone cannot advance far unless brought in touch with others in the same field. It may be noted that the opportunity is peculiarly advantageous for those teachers in smaller colleges who can make use of a corps even of un- trained assistants in the collection of various data. We feel it a privilege to invite kindly criticism of this report and sug- gestions as to the best means for carrying out the aims in view and for securing the co-operation of the largest number of workers. (Signed) A. E. Brree, Chairman. C. H. EIGENMANN. C. A. Kororp. G. C. WHIPPLE. H. B. Warp, Secretary. Vea i’ JACOB DOLSON COX NECROLOGY. JACOB DOLSON COX, OF OBERLIN, OHIO. Jacob Dolson Cox was born at Montreal, Ontario, October 27th, 1828. His father, a resident of New York City, and an architect by profession, was then living in Montreal where he was engaged in supervising the construction of the roof, etce., of Notre Dame Cathedral which he had designed, and upon the completion of which he returned to his home in New York City, where the boyhood years of the subject of this memoir were passed. In 1846 Jacob Dolson Cox entered Oberlin College at Gberlin, Ohio, from which institution he graduated in 1851, receiving the degree of Master of Arts. In 1877 the degree of Doctor of Laws was conferred upon him by Yale College. He married Mrs. Helen Finney Cochran, widow of Prof. William Cochran and daughter of president Finney of Oberlin College, and in September, 1851, he accepted the position of superintendent of schools in Warren, Ohio, which position he held until 1854, and in the meantime stud- ied law and was admitted to the bar. In 1859 he was elected to the senate of Ohio, in which he served during the years 1860 and 1861. Upon the breaking out of the civil war and almost immediately after the firing upon Fort Sumter, he was appointed by Goy. Dennison, Brigadier General of Vol- unteers and was placed in charge of Camp Dennison, near Cincinnati, which was the great receiving and distributing camp for volunteers, and while in command of the camp he displayed great efficiency in the organization of the troops and in all the duties connected with the responsible position which he held. In July, 1861, he was assigned to service in the field, drove Wise out of the Kanawha Valley, West Virginia, and took a prominent part in the battles of South Mountain and Antietam, in which he commanded the 9th army corps. In the Atlanta and Franklin and Nashville campaigns Gen. 198 JACOB DOLSON COX Cox commanded the 23d army corps, and in 1864 he was commissioned Major General of volunteers. In 1865 Gen. Cox was elected governor of Ohio, in which office he served during the years 1866 and 1867, and at the ex- piration of his term moved to Cincinnati and resumed the practice of law. In March, 1869, Gov. Cox was appointed Sec- retary of the Interior by Pres. Grant, and filled this position with distinguished credit until November, 1870, when he re- signed, and returning to his home in Cincinnati, Ohio, again resumed the practice of law. Soon after assuming his portfolio in Pres. Grant’s cabinet, Goy. Cox was confronted with the practice then prevailing, of levying assessments upon the department officials and clerks for political purposes. To this Gov. Cox was inflexibly op- posed, and was supported in his opposition by Pres. Grant, who assured him that the practice should no longer be en- forced in the Interior Department. In September, 1870, how- ever, during his temporary absence from the capitol, certain prominent politicians collected assessments from certain em- ployes in the Interior Department, greatly against the will of the employes. Upon learning of this, Sec. Cox became highly incensed and resigned his portfolio, taking the ground that if his views could not prevail in the conduct of the Interior De- partment he would have nothing to do with it. His resigna- tion was reluctantly accepted by Pres. Grant, who personally sympathized with the views of Gov. Cox, but seemed to have been unable to control the clique of eminent politicians, who, themselves holding high offices under the government, were fully in accord with the system of political assessments. In 1873 Goy. Cox was appointed president of the Toledo and Wabash Railroad Co. and removed to Toledo, Ohio. He re- mained as president and receiver of this R. R. until 1878. In 1876 he was elected to congress from the Toledo District, and served as Congressman until March, 1879. Upon the expira- tion of his congressional term Goy. Cox retired from politics and thereafter declined to take any active part therein. In 1878 he removed to Cincinnati, soon after becoming Dean of the Cincinnati Law School, which position he held until 1897, when he resigned his position, retired from active business and removed to Oberlin, Ohio, where he resided until his death, which occurred at Magnolia, Mass., on August 4th, 1900. JACOB DOLSON COX 199 He was president of the University of Cincinnati from 1885 to 1889. It was about the time of his removal to Toledo that Gov. Cox became interested in microscopy, which, as many other eminent microscopists have done, he first adopted as a recrea- tion. His first microscopical studies were upon fresh water forms, particularly in the class rotatoria; but as might natur- ally be expected he speedily became interested in the study of the diatomaceae, to which his chief attention from that time forward was given. In his microscopical studies Gov. Cox displayed that painstaking thoroughness and striking power of logical analysis which he displayed in all his works in every walk of life. Almost at the beginning of his work with the microscope Goy. Cox adopted a system of keeping notes of his work and observations, which eventually grew to remarkable proportions. Among his belongings which he left at his de- cease, were some twenty or more note books completely writ- ten and filled with the records of observations, drawings and notes of his conclusions and deductions, all carefully dated and neatly written out, which work alone would seem to have been sufficient to absorb all the leisure time that one man would be able to devote to such work. In addition to this, however, he had made an almost complete card catalogue of the diatomaceae comprising the nomenclature and synonymy of each species, references to leading works in which it is fig- ured and described, and Maltwood finder references to speci- mens of that particular form in slides among his own collec- tion, which numbered some thousands of slides. In addition to contributing articles at intervals to the microscopical jour- nals, Goy. Cox contributed several important papers to the Proceedings of this Society. In 1874 Gov. Cox took up the subject of photo-micrography, and in 1875 began making the series of photo-micrographs of diatomaceae so well known to many members of the society. Of this series of photo-micro- graphs, several hundred in number, Goy. Cox kept the same careful record as of all his other work with the microscope. His note books show the date when each negative was made, will full particulars concerning the object, illumination, ap- paratus employed, photo-micrographic technique, etc. Prints from this carefully numbered series of negatives were distrib- uted by him to all the leading authorities on the diatomaceae. As a result of this work Goy. Cox received at the Antwerp ex- 200 JACOB DOLSON COX position in 1891 the gold medal awarded for the greatest ex- cellence in micrography. In 1881 Gov. Cox was elected a fellow of the Royal Micro- scopical Society, and in August, 1882, was elected a member of this society at the Elmira meeting. At the Chicago meeting in 1883 he was present and contributed a brief paper descrip- tive of the modification devised by him of the Wales Micro- scope stand, which modification became known as the “Con- centric” stand. At this meeting he was nominated and elected president of the society for the next meeting, which was held at Rochester in 1884. His presidential address at the Roch- ester meeting embraced a masterly discussion of the Tolles- Wenham controversy over the angular aperture of objectives used with balsam mounts. No one who had followed that lengthy and acrimonious dispute can fail to admire the clear, logical and exhaustive, yet brief, analysis of the arguments pro and con and the very numerous errors so strenuously maintained by the opponents of the “extra” balsam angle dur- ing the years when that unfortunate controversy dragged its- slow length along through the literature of microscopy. At the same meeting Goy. Cox read an interesting paper on photo- micrography with high powers, which displayed not only a thorough acquaintance with the subject and great experience in the work, but served as a text around which animated dis- cussion was waged for the next two or three meetings, the views maintained and the practice advocated by Gov. Cox in that paper being finally vindicated by demonstrations afforded by the work of several other well known and active members of the society. At the Cleveland meeting in 1885 Goy. Cox read a second paper on the subject of photo-micrography with high powers. At the Chautauqua meeting in 1886 Goy. Cox, in collaboration with Mr. W. H. Walmsley, conducted a prac- tical exhibition of photo-micrographic work which was of great value in familiarizing many interested members of the society with the technique and manipulation essential in that class of work, which at that time was rapidly growing in favor. At the Washington meeting in 1892 Gov. Cox was a second time elected president of the society, and presided at the Mad- ison meeting in 1893. In the intervening years he contributed to the society other valuable papers, one of the most painstak- ing being his paper read at the Detroit meeting in 1890 on the classification of the diatoms. After 1895 he practically ceased JACOB DOLSON COX 201 work with the microscope, fearing its effect upon his eyes, but his interest in microscopy and the study of natural history by the aid of the microscope continued unabated during his life. His accumulation of careful and minutely detailed records, not only served as valuable helps to Gov. Cox in his work, but afforded the means by which the path he followed and the steps he took in his investigations can be accurately fol- lowed and fully understood by one who now reads the same. During the last few years of his life Gov. Cox in company with his son, J. D. Cox, Jr., of Cleveland, spent several weeks each summer in yachting upon the New England coast, some- times accompanied by other relatives and friends. During these periods Gov. Cox kept a careful log of their daily wander- ings, noting the barometer reading, course of winds, distances sailed, points visited, etc., with all the careful attention to de- tail which characterizes the most experienced ship master. His wide experience as an officer combined with his habits of exhaustive research and his power of keen analysis fitted him especially for the work of a military critic and historian, in which Goy. Cox attained a high repute. His reviews of military works in “The Nation” alone would fill several vol- umes and are marked by charity, lucidity and original contri- butions to history and the science of war. He wrote the arti- cles “Atlanta” and “The March to the Sea,” “Franklin and Nashville” for the “Scribner Campaign Series” in 1881 and 1882; several articles for “Battles and Leaders of the Civil War” published in 1884-1888; “The Battle of Franklin,” of which magnum pars fuit, in 1897; and “Military Reminiscences of the Civil War” in 1900. He also prepared an admirable analysis and exceedingly clear exposition of the facts in the Fitz John Porter Case, which he published in 1882, during the time when persistent efforts were being made in congress for the rehabilitation of General Porter. In the outset of his studies at Oberlin College Gov. Cox had intended to enter the ministry, and he pursued his theological studies for some time. Eventually, however, he seemed to deubt, probably without sufficient reason, his fitness for the ministry and gave up the idea of a clerical career. It is likely, however, that his theological studies accentuated and intensi- fied the natural broad christianity of his character, which dur- ing all of his varied and illustrious career was one of the potent features of attraction which so deeply endeared him 202 MOSES CLARK WHITE to all who became well acquainted with him. His personality was attractive and none who met him can fail to remember his tall and striking appearance, his rich sonorous voice, and the peculiarly frank and friendly expression of his face. C. M. Vorce. MOSES CLARK WHITE, A. M., M. D., or NEw HAVEN, Conn. By the death of Professor Moses Clark White, M. D., which occurred October 24, 1901, the American Microscopical Society lost one of its oldest and most enthusiastic members. Dr. White was born in Paris, N. Y., July 24, 1819, and was accordingly over eighty-one years old at the time of his death. His preliminary education was obtained at Cazenovia Sem- inary, Cazenovia, New York, after which he entered Wesleyan University at Middletown, Conn., in February, 1842, and grad- uated third in rank in the class of 1845. The following two years he spent in New Haven, Conn., in theological and med- ical studies at Yale University, meanwhile preaching in New Haven and adjoining towns. Having been ordained an elder in the Methodist Episcopal Church, and soon after this appointed as a missionary to China, he sailed together with his wife from Boston on April 15, 1847, going to Foo-Chow, and the years from 1847 to 1853 were spent in medical missionary work in that country. His experiences in those early days of Chinese missionary work were most varied and unusual, combining a large dispensary service with a private practice that covered a wide territory. During this time, he also published a translation of the gospel of Matthew in the colloquial language of Foo-Chow, which is said to have been the first book ever published in that dialect. After seven months’ residence in China Mrs. White died, and Dr. White’s own health finally became so impaired that he was compelled to leave there in 1853, and he returned to New Haven. Soon after his return, he published “An Introduction to the Study of the Colloquial Language of Foo-Chow.” In 1851, before his return from China, he was again married, his wife being Mary Seely, of South Onondaga, N. Y. Upon his return to this country he again took up his med- ical studies at Yale and graduated in medicine in 1854, then MOSES CLARK WHITE MOSES CLARK WHITE 203 establishing himself in New Haven asa physician. He at once became interested in microscopy, and in 1857 was appointed ‘lecturer on microscopy in the medical department of Yale University. Ten years later, he was advanced to the newly established professorship of Microscopy and Pathology in the same institution; and this position of Professor of Pathology he retained until a few months before his death, when he severed his active connection with the Medical School and was elected Professor Emeritus in the same subject. Aside from his duties as a practicing physician, and as a teacher of medicine, Dr. White found time to act as an in- structor in Botany in the Sheffield Scientific School from 1861 to 1864, and he was also a lecturer on Vegetable and Animal Histology in Wesleyan University, his old alma mater, from 1869 to 1875. For many years he was connected with the New Haven Hospital, as one of the attending physicians, and as pathologist, at the same time being a member of the board of directors of that institution. He was also secretary of the Connecticut State Medical Society from 1864 to 1876. Since the creation of the office of Microscopist of the New York Medico-Legal Society, that position has been filled by Dr. White. His connection with and work for the American Microscopical Society since he became a member of it in 1885 is too well known to need repetition here. Always an active member in the full sense of the word, he served in 1898 as Vice President, and would have been honored with the posi- tion of President lad he permitted the use of his name. From the establishment of the office of Medical Examiner, or Coroner’s Physician, in Connecticut in 1883, until the time of his death, Dr. White held that position for the town of New Haven. His success in this office is perhaps best given in the words of the Coroner of New Haven County: “As Med- ical Examiner, he has been associated closely with me for the past seventeen years, during which time he has never wavered in the complete performance of his duties, rendering honest, capable, reliable, efficient, valued service. As Coroner of New Haven County, State of Connecticut, I have twenty- five Medical Examiners connected with the office. Dr. White was the peer of them all, the one to whom I went and whose knowedge and counsel I availed myself of.” Besides the publications already referred to, Dr. White as- sisted largely in the preparation of “Silliman’s Physics,” and 204 MOSES CLARK WHITE wrote the chapter on Optics. He revised and edited the sec- ond edition of “Porter’s Chemistry.” For some years he has shown great interest in micro-photography, and in the exam- ination of blood stains, not only by chemical means, but par- licularly by measurements of the red blood corpuscles. This Jatter method he emphasized as of value in differentiating the blood of various animals. His publications in this line have been many and important, and the members of the American Microscopical Society have had an opportunity to see the ex- tensive and exact nature of his work. More recently he has devoted much time to the perfection and use of the projection microscope. This incomplete outline of the life-work of Professor White may give some idea of the energy of the man and of his abil- ity in various directions. But only those who knew him best could rightly appreciate his true worth. He was so quiet, so unobtrusive in his manner, that a close acquaintance was necessary to really know the man. And such an acquaintance could not fail to show his uniform kindness to all, his remark- able enthusiasm which kept him to the very front in his work in microscopy, and a determination which kept him active al- most to the day of death. One of his long-time friends has well written of him, “Ear- nest in all his undertakings, of the highest integrity and up- rightness of character, enjoying the respect and confidence of all with whom he was associated, his memory deserves to be honored by some pen that will thoughtfully, truthfully, and lovingly delineate the meritorious and estimable char- acteristics of Dr. White as they were known to those who knew him best.” C. J. BARTLETT. New Haven, Conn., April, 1901. PROCEEDINGS OF The American Microscopical Society MINUTES OF THE ANNUAL MEETING HELD IN NEW YORK CITY, JUNE 28, 29, 30, 1900 First SEssion TuurspAy, June 28 The twenty-third annual meeting of the Society was called to order by the President in Schemerhorn Hall, Columbia University, New York City, at 2 ep. m., Thursday, June 28, 1900. After the reports of the Curator, Secretary, and Treas- urer had been read, it was, on motion, decided that owing to the very early date of the annual meeting, the financial year of the Society should be extended to October 1st rather than close at the date of the meeting. The Custodian was directed to send to the Smithsonian Institution a set of back volumes save Vol. VI. The report of the Limnological Committee was given in- formally, and it was decided that the opinion of the com- mittee with reference te an increase in number was correct, and that a chemist, a physicist, and a bacteriologist should be added as soon as suitable members could be obtained for these fields. The committee was given time, and in- structed to find those willing and able to undertake the work. The Society then voted unanimously to accept the invitation of the Board of Directors of the New York Zoologi- cal Garden to visit the same with the Zoological Section of the A. A. A. S., and thereupon adjourned to proceed to the Zeological Garden, where a most enjoyable afternoon was passed in inspection of the plant under the leadership of Prof. H. F. Osborn, of Columbia University, and Dr. Wm. T. Hornaday, Director of the Garden. 206 PROCEEDINGS OF THE SECOND SESSION At 8 p. M. the Society convened in the rooms of the New York Microscopical Society, 64 Madison Avenue, and listened to the annual presidential address by Dr. A. M. Bleile, on “The Detection and Recognition of Blood.” At the close of the meeting an informal reception was tendered the Society by. the New York Microscopical Society, and members of both Societies spent some time in social enjoyment. THIRD SESSION Fripay, June 29 The Society was called to order at 10 a. M., in Schemerhorn Hall, and after the report of the Executive Committee for the fiscal year had been read, the President appointed Mr. John Aspinwall and Dr. M. A. Veeder Auditors of the Cus- todian’s report, and Mr. Magnus Pflaum and Mr. C. C. Mellor of the Treasurer’s report. A nominating committee was then elected, consisting of Messrs. Gage, Kuehne, R. H. Ward, White, and Whipple. The Society then listened to the read- ing of the following papers: “Photographing the Spectra of Colored Fluids,” by Dr. Moses C. White, of New Haven, Connecticut. In discussion the President made extended reference to his address of the previous evening in connection with the paper in hand; the latter was also discussed by Professor Gage. A paper on “A Method for the Measurement and Demon- stration of the Size of Minute Bodies” was read by Dr. Henry B. Ward, and discussed by Prof. S. H. Gage and Mr. Magnus Pflaum. “The Life-Work of Herbert Spencer” was reviewed by Mr. H. R. Howland, and further tribute to his work was added by Dr. R. H. Ward and Prof. 8. H. Gage. The Society also discussed generally the Spencer-Tolles Fund, and voted that the President should appoint a com- mittee of three, who, together with the Custodian, should be especially charged with the completion of the fund before the next meeting. A paper on “Methods in Embryology” was read by Prof. S. H. Gage, and discussed by Dr. H. B. Ward; and a second paper on “The Comparison of the Development of the Larynx in Toads and Frogs” was also read by Professor Gage and discussed by Drs. L. Schoeney and H. B. Ward. The Society then adjourned for dinner. AMERICAN MICROSCOPICAL SOCIETY 207 FOURTH SESSION The Society was called to order by the President at 2:50 Pp. M., and the Report of the Limnological Commission was presented by Dr. H. B. Ward, Secretary of the Commission. A paper “On the Distribution of Growths in Surface Water Supplies and the Method of Collecting Samples for Examination,” by Dr. F. S. Hollis, was read, in the absence of the author, by Mr. G. ©. Whipple, and informally dis- cussed by a number of the members and visitors. A paper on “The Necessity of Maintaining a System of Field Work on Surface Water Supplies,” by Mr. H. N. Parker, was read, in the absence of the author, by Mr. G. C. Whipple, and discussed by the President and others. A paper on ‘The Cladocera of Nebraska,” by Dr. Chas. Fordyce, was read by the Secretary in the absence of the author, and informally discussed. “The Work of the Mount Prospect Laboratory of the Brook- lyn Water Works” was presented by Mr. G. C. Whipple, with lantern slides illustrating the laboratory and its equipment and work, and was discussed by Messrs. John Aspinwall and H. B. Ward. A paper on “Some New Forms in the Cave Fauna of Mam- moth Cave” was read by Dr. C. H. Eigenmann, and discussed by Messrs. John Aspinwall, H. B. Ward, and others. A paper on “The Modern Conception of the Structure and Classification of the Desmidiaceae” was read by title in the absence of the author, Prof. Chas. E. Bessey. A paper on “Some North American Hydrachnidae, hitherto undescribed,” was read by title in the absence of the author, Dr. Robert H. Wolcott. A paper on “Limnological Studies at Flathead Lake” was read by title in the absence of the author, Prof. M. J. Elrod. The committee on the Spencer-Tolles Fund was then an- nounced by the Chair as H. B. Ward, Adolph Feiel, and H. R. Howland. The Society then adjourned. FIFTH SESSION SaTURDAY, June 30 The Society was called to order by the President at 9:30 A. M. A paper on a “Method of Reproducing Color Effects of Stained Sections in Lantern Slides” was read by Mr. John Aspinwall, and discussed by Dr. H. B. Ward. 208 PROCEEDINGS OF THE A paper on “Some Hints in Bibliographic Work in Science” was read by Dr. R. H. Ward, and informally discussed. A paper on “A New Ear Fungus of Man,” by Dr. Roscoe Pound, was read by title in the absence of the author. A paper on “Methods in Staining and Killing Protozoa” was read by title in the absence of the author, Prof. M. J. Elrod. A paper on “Synthetic Alcohol as a Fixing Agent for Tissues” was read by title in the absence of the author, Dr. T. E. Oertel. The Nominating Committee reported the following choice of officers for the ensuing year: For President....Prof. C. H. Eigenmann, Bloomington, Ind. For Vice-President..... Mr. Chas. M. Vorce, Cleveland, Chio. For Second Vice-President... . 2. 0... oh. sre ee BYR ENS Seed aoe ops Mr. Edward Pennock, Philadelphia, Pa. For Elective Members of the Executive Committee...... Mr. John Aspinwall, New York City. Dr. C. A. Kofoid, Urbana, Ill. Dr. A. G. Field, Des Moines, Iowa. A yote of thanks was passed to the officers of the Society for their services, to Columbia University for the use of the rooms and apparatus placed at the disposal of the Society, to the President and members of the New York Microscopical Society for their cordial hospitality, and to the Board of Directors of the New York Zoological Garden for the courte- sies shown members of the Society during the visit to the garden. After appropriate words of thanks to the Society, the re- tiring president called the new incumbent of the office to the chair. On taking his position, the President, Dr. C. H. Eigen- mann, expressed his thanks to the Society for the honor con- ferred upon him, and the hope that the coming year might be more successful even than the past. He then declared the meeting adjourned, the place and date of the following meet- ing to be decided by the Executive Committee. Henry B. Warp, Secretary AMERICAN MICROSCOPICAL SOCIETY 209 TREASURER’S REPORT FROM AUGUST 15, 1899, TO SEPTEMBER 20, 1900 To Membership dues, 1898, 1 ............. To Membership dues, 1899, 15 ............. To Membership dues, 1900, 1693............. To Membership dues, 1901, 33 ............. fo Membership dues, 1902, I) 2.0.5 5.05--- fo; Admission: fees, 1900, 22.02.2020 222 shes PorAdmission fees; 1901), G2 -\.i56 62.0 .< e010 0% Po SHRP seEIDErs,:"V OL) Nek ss oa e's alcletoite a oo ste He snpseripers,) Vols KML. Sut ses voce ot Nees Mo Sale af Proceeding 3). cei «/astas cree. os as PorAdyertising.) Vol.) 2X oo i iselescwseces Hers ovmdvertisine. Vols! XOMD. . lic. ciniele ec ece ene To Balance due Treasurer............. Expenses Columbus Meeting................ Pay COMING. CCLCLATY ... 5 945 Niagara St., Buffalo, N. Y. BP RMSOHEE WV; cE.5): (Odreberatein sfsteiarelere's vichsjsieelstel Box 1033, Rochester, N. Y. DunuaM, FE. K., M. D., 92....338 E. Twenty-sixth St., New York City EASTMAN, Lewis M., M. D, ’82...772 W. Lexington St., Baltimore, Md. EIGENMANN, Prof. C. H., ’95..University of Indiana, Bloomington, Ind. Parrort, Prot. ARTHUR EH .. 7 Odie e316 2 4 Irving Place, New York City PRETO E WTR. Bl, 798). 42 ssi cise dete 4 Fulton Ave., Rochester, N. Y. HEROD) Prot. MORTON Ji. M: A., Be A MOUS.) 79852 ac is sleralale nie Bees hse Seater ol et: cas vaetayala seaheve erator skehsiar ats 226 S. Fifth St., Missoula, Mont. SERS JOHN; Mi. Dr 83ic sas sie sellers « 1014 Fourteenth St., Denver, Colo. PEMA Tels PAN 2 Nea) O's es arcs okorecesotovevet stalcrars 16 Pearl St., Council Bluffs, lowa EWELL, MARSHALL D., LL. D., M. D., ’85...... 59 Clark St., Chicago, Ill. IB eOHIN, (VVio is, Min Dee Mey Sr Hs cEesy Min Soir O 0 eyereierelarclaielat-"elat<1<1- ; ..Embankment Chambers, Villiers St., London, W. C., England ein AMOnMP HE Mis ID) Sill fev scceretarelel hers 520 E. Main St., Columbus, Ohio ding Errors IM IDA ne dee WE Sey eritincacio cc 72 Niagara St., Buffalo, N. Y. ThraresO ish COTS SPI DN UR IN ETS dsiic Gouodoe Uonocoue Fairbanks, Florida BERBIS. Prot. HARRY: Bi 79G6). <0 oi 118 York St., New Haven, Conn. Bene Ae Ge Mig) fee ole: stele)'slel ofaleieistake Summit Place, Des Moines, Iowa IGN ERAN SiRsy Mi. IDS 798 clos o/ccle elec «reel 2 Union Place, Troy, N. Y. SERBS MUACKS OB'5. 5) Jslr ee clears: sieleie's Zeiss Optical Works, Jena, Germany IE SEDER ERE Va iS LORE aye ss| 299 |e cc tuie ore, 5 elaieie ste seis ees @ sie Pleasantville, Ohio Print, JAMES. M., M. D., ’91...... “The Portland,’ Washington, D. C. HOD MOR OHARTHMA Hise) Ss4 Ay My, Ente 1.572985 5 oi <(orac etalciels aie) saietelel a) « Hapstate sie! Nebraska Wesleyan University, University Place, Neb. HOSTER. EDWARD; 99) 5 ..\22.0/2 6 «le <= 0 P. O. Box 405, New Orleans, La. OX IORCARNCH 927 se caeadsadese U. S. Patent Office, Washington, D. C. PE re Acting ee elie eae Ie DG AG) 312s, 6, Lave ov alielol siiere 342 Ohio Ave., Columbus, Ohio Hicrm Bree EPA So) Gres Mis Dene Blot ERs OME US. 50S ot cores e) ce ye)elalels|are/~\als/astat sl « Sa Gatien a teak ae nD Orn HONE ICE 38 Central Music Hall, Chicago, U1. GAGE, Prof. Simon H., B.S., 782... ..% Cornell University, Ithaca, N. Y. GAGE, Mrs. SUSANNA PHELPS, *87........ 216 South Ave., Ithaca, N. Y. GREE Se MMU MBB IGA se. eca.c-sncho.siee.s PAD Yeu WE Lanne tay dns Chevy Chase, Md. GOODRICH Wis Eke NTS Ds. 798i... siocle/siwleie's/o/e) aleiciolsislele,eie\ela's ts Augusta, Ga. GROSSROPEURNEST) Coy) Me IDs) 799i crave) eiciw ls ol cle eleleleiele Wauwatosa, Wis. EPAAG iba Mi De) SOs ee sles acieiele 1121 Washington St., Toledo, Ohio PEDAIVA NEAT Oo. Hee Hep buen ME a1 Darr lala: stares c\erels 103° First St., Troy, N: ¥. Hanks, Henry G., F. G. S., °86..1124 Greenwich St., San Francisco, Cal. VATED, JOHN). 1-55 (G2. <0 <0 - 333 Arsenal Ave., Indianapolis, Ind. IEDER UZ. VAR TENOR y Aree. DS. 29615 ooo: a Hel ot chord clct ch che) sh aha) ates Halstead, Kan. TeineveTm fe Mana led Deis Sram ene AAG oc 20 Court St., Cortland, N. Y. i ERE ERT OMe DEA Biilay ayer ahslelee) cic 24 High St., Buffalo, N. Y. HOREMAN; JOS) Hy MoD: 796% \2)2 6.0 i afore o's 111 Steuben St., Pittsburg, Pa. 218 THE AMERICAN MICROSCOPICAL SOCIETY Howeroox, M. L., M. D., ’82....43 E. Twenty-first St., New York City HMOLnIS) (PREDERICGK)\ Se PS DOO cree Pc clk ta tealahcke le telaveve ate eters ietetiote BD Ae (ve Rael e Peles eral al ene a Netarets amis te Yale Medical School, New Haven, Conn. OEMS. ALN MEDS PSB. ic cick als isis eis 205 Jackson Block, Denver, Colo. HTOSEAINS) (WME TOE cls ecieislovciele’e Room 55, 81 S. Clark St., Chicago, Il. HOWARD, CurTIS C., M. D., ’83...... 97 Jefferson Ave., Columbus, Ohio. HOWE UW. EEE PhD OO es ince slots s scerersiclalsietelaere Gielen Evansville, Ind. HowLAND, HENRY R., A. M., ’98........ 217 Sumner St., Buffalo, N. Y. BOMPHREN Prot kObN DE ees. Ob lever niece lelsle reine te lolstotoeletetaeiereneiis bys Wislkicre mielsvelelaiate sieiatela etee ete) & State Normal School, Jamaica, N. Y. AVIA TY OG LED LS ha tcyare are tain cvevekele rete 69 Burling Lane, New Rochelle, N. Y. JACKSON, DANIEL Dana, B. S., ’99....177 Sixth Ave., Brooklyn, N. Y. VARHS? BUSHROD Wis IM DE 7G cos ciere veleteiers elevetele. ste tolokecie Cyateneteteteanie Sass eters N. E. cor. Green and Eighteenth Sts., Philadelphia, Pa. AMES: CHURANK, 2 Het Dis Me DBF oe ts 3032 Ohio St., St. Louis, Mo. AMES OSU ees (Ore siciere soles ie ois bree iere mart rastoecs 108 Lake St., Chicago, Tl. JOHNSON, FRANK S., M. D.,'’93.........- 5221 Prairie Ave., Chicago, Il. SFOEINGONS VV Me OLD sie IDs. COR Nie, alate crelreevete ioe taferetererelocerereheretete Batavia, N. Y. JOHNSTON, Tava DL ME DPOB six cta ever elo ters fals teicrsiaysvelel oie eet Whittier, Cal. JoneEs, Mrs. Mary A. Drxon, M. D., F. R. M. S., 798............ Vet PA Petey 249 EK. Eighty-sixth St., New York City SUD Asan COHAN CIGY20 POO creole sictsicle eis seicte)oichal state 412 Lake St., Madison, Wis. GTO GG see DDT Gum srotchetoiess aieie lei cresereistenere & cles Battle Creek, Mich. KERR, ABRAM TUCKER, JR., 95.......... 1386 Maine St., Buffalo, N. Y. KINGSBURY, BENJ. F., A. B., M. S., 98..125 Dryden Road, Ithaca, N. Y. RRP A MRTG Ae hes iO Sacral dere o cre ciclas shetein er esave, ota aroueveters Springfield, Ohio Kororm,; (CaAmnns jAs PhS Dis 798,050 a's iste aie sisls ih inletels ne eee Ae SRL UA Re lole & ejateiehe BESIE LA is University of California, Berkeley, Cal. TO TAT AL AT Ms SPL O MEE ez Lal feletelishs fav ib Fane fous 32 S. Fourth St., Easton, Pa. KRAR ET [WIRD LAMS 795.2) 32). 411 W. Fifty-ninth St., New York City Krauss, Wm. C., B. S., M. D., ’90....371 Delaware Ave., Buffalo, N. Y. GUAEELNED: ABIWVPRO S ia is areyeelers eisialeiie ies 19 Court St., Fort Wayne, Ind. SEAMEN.) MDS POS Secret ss lela 910 T St., N. W., Washington, D. C. LANDSBERG, AG! STO Mic Keltetetels where 156 Woodward Ave., Detroit, Mich. ATA MISS VES MEAD SS DYED SE Se BR, UMS.) 788i. cipele sale everett wihiiaielets wk eisai eras 808 Morse Ave., Rogers Park, Chicago, Il. LAWTON, HED WARD (be. OGcinete ce cesslelsionrcre sie.u's 3 Linden Ave., Troy, N. Y. LEIPPE, J. HARRY, °96........ Second and Franklin Sts., Reading, Pa. Wis; VERS: KAT HARENE IBS S289)5 5) o's 66 cieiwlaleieosevevereisicls cyaieioteuelenaeeieene at Sus iene ieils tak obs otaiedsh ers & “Elmstone,” 656 Seventh St., Buffalo, N. Y. TURSWAS, ELBA WESC Sir leis is oie siaucte oetekate etd lekeiere 408 So. Galena St., Dixon, Il. LING. de LDWARD. D. IDSs OSes sre tcle le 39 State St., Rochester, N. Y. LOOK: SOHN Da OSck lace either arate tc le aig [eter P. O. Box 129, Haverhill, N. H. LOMB ADOUPH, | 92/0 \aGieticis terior selec s' 48 Clinton Place, Rochester, N. Y. THE AMERICAN MICROSCOPICAL SOCIETY 219 EO AME EDN RY: ABA Lik Livtale. siclacehaiel ata feb ro 48 Clinton Place, Rochester, N. Y. Loomis, CHANDLER H., ’87..Atlantic Dredging Co., 31 Pine St., N. Y. City Love, Prof. E. G., F. R. M. S., ’91..80 E. Fifty-fifth St., New York City Lyon, Howarp N., M. D., ’84....... ary ay St 103 State St., Chicago, Ill. NAINTION:, Wie. Pe, Mis Do 7B Biosci cienyers 32 W. Adams Ave., Detroit, Mich. MARSHALL, CoLLins, M. D., ’96....2507 Penn. Ave., Washington, D. C. PASS EVA TL 7 WV Mick) ed Bin.) 9S stl svcic seve S elajeielpiovslalaieteraislcies flo snare Coudersport, Pa. ACME AEA NS! HRT MESR JHS.5) °29'f)a1d/c)apaleielaye) ey «)a/ePoteterey sicheletoteye > New London, Ohio MOGALTA ALPERT, bo WDE i480 wialdsicis)s jie euskoniole nie eieleisveyoistevstetetele atatel= Neen 414 Monadnock, Dearborn and Jackson Sts., Chicago, III. iGIKCACy, TS OSHP EH: 7844 wy.) o2 ce ate elersiedenere teleichenels 259 Highth St., Troy, N. Y. McKim, Rev. HASLETT, ’85......9 W. Forty-eighth St., New York City McMILLEN, R. M., M. D., ’00.......35 Twentieth St., Wheeling, W. Va. MEADER, LEE Dovuctas, M. D., °96..2651 Gilbert Ave., Cincinnati, Ohio Mrrmnony CHAS OC... Shs cennsste tetas) ceases 319 Fifth Ave., Pittsburg, Pa. MEP GHR A sCEEnnORD:) Me Din ein IMIS 282) )-lanics eteperelelelereisieimlels Sayed ie.c fatal ava miayercfetelotart istets fetes 324 Montgomery St., Syracuse, N. Y. Mnronre MRMDMRECK Wes ME. Dl He ReMi Sit 83 Sole. cise sisielelsieise sts EWEN anes ey ahicy ps Stagat ovate euch e feheonedaneraiepenek sratey’ 2540 Prairie Ave., Chicago, Il. MR OR ER PE MOiin DV shee tie 99 eet atetoratetom affel aleyst ol etsyelars, ole oll seleiee ee eee 261 % OOMS TUEILUL OI lew levsin\nin nia talo te iaveleleye a ialeleterontersieisrele’slelersioieis eioinielet atten 265 COMSUUEILELOT ieichersls! srelelelstere esetereisiole toys) sleieleleye. eats ioharate alos te) eisteh ete te iateteeataeet 211 *Consumption and Cancer, Defective Development and Disease, with Special Reference to the Curability of, by M. A. Veeder.. 17 COX TACOD SDOISOM TS 220 sw ierdeors a ieiato levels, aes bos latetelste)aierbife a eel Sete ReteCRae Renate 197 Custodian’s Report, Spencer-Tolles Fund.................-eeceeee 210 PDAVICS, (eo) OLE UO CMC yeteleisic/elaieis ais eieieloieis(oieys)elalelot=\a(e) stele) oVolstel etal tatete enemas 249 *Davis, Ellery W.—Obituary Notice of John Eugene Davies....... 249 *Defective Development and Disease, with Special Reference to the Curability of Consumption and Cancer, by M. A. Veeder... 17 Desmids, The Modern Conception of the Structure and Classifi- catlonsol, oy Charles) IMs ABESSCYirleil< > cl «lero leis ele siete ee tere keine aici 89 Description of a New Cave Salamander, Spelerpes stejnegeri, from the Caves of Southwestern Missouri, by Carl H. Eigenmann... 189 Description of a New Genus of North American Water Mites, with Observations on the Classification of the Group, by Linajorermne Els Wolo 5.415 00600 SOSUO OOOO IR GON Ado Oo ba0G Sho Sonnds so. 105 Detection and Recognition of Blood, by A. M. Bleile............. ct *Diabetie Blood, The Reaction of, to some of the Anilin Dyes, by We As Lilia ti ain 9, feos laicioieiwlelove alcinlors wlateweie cvs Sols tetulele kote teiovaietete ete ieteieae 31 *Diatoms, The Modern Conception of the Structure and Classi- fication of."by) (Charles iH: (BESSEiis ccc. otele\sis)>\)is/= lee feleiel=te eteemetenerae 61 EDowbled ary, evemriy. HE2)./si0jstale\ieje\siele » (ofelale,«\ele\e as)s>(alolele evel felete se eileen ites 250 *Eigenmann, Carl H.—Eyes of the Blind Vertebrates of North America. II. The Eyes of Typhomolge rathbuni Stejneger..... 49 Eigenmann, Carl H.—Description of a New Cave Salamander, Spelerpes stejnegeri, from the Caves of Southwestern Missouri 189 Elrod, Morton J.—Limnological Investigations at Flathead Lake, Montana, and) Vicinity, dialliy.. 1S99o oy. ./iic/cnrc uo leleinlolotelateielaiaentels 63 *Expedient for Use in Difficult Resolution, by R. H. Ward........ 111 *Eyes of the Blind Vertebrates of North America. II. The Eyes of Typhomolge rathbuni Stejneger, by Carl H. Eigenmann 49 *Ferris, H. B.—Obituary Notice of Albert E. Loveland............ 251 Flathead Lake, Montana, Limnological Investigations at, and Vicinity, July, 1899, by Morton J. Hlrod.............cseccesrne 63 INDEX 225 Fluids, Photo-spectography of Colored, by Moses C. White....... 99 Fordyce, Charles—The Cladocera of Nebraska.................2: 119 *Gage, Simon Henry—Some Laboratory Apparatus............... 107 Hollis, Frederick S.—On the Distribution of Growths in Surface Water Supplies and on the Method of Collecting Samples for EP AMINTV A GLOW ssi valores He belek e Chetesierm, elarevorete slave stcley uals chair sicusieioietetercbevanets 49 *Howland, Henry R.—Obituary Notice of Herbert R. Spencer..... 252 Human Ear, An Addition to the Parasites of the, by Roscoe ESORIIT RUG oict oe: sais e = 8 svoiaiaiia wlek peters miciehens(erarahanetave) sietalc eb srasa) crave al sve en eme ISS 81 *Hydrachnidae, New Genera and Species of North American, by Paemet rl. WoOlGOtt! once cepiacia sisi osiacis iw a laarcrolalerstern stotartate alyee *Incubator for Student Use, by Veranus A. Moore................ 103 *Infusoria, Notices of Some Undescribed, from the Infusorial PAIN AOL MOUISTANA, syed Oay OMMTb I ala aiels wale sla) oclelsiele oie \ nla) aleve (cies 87 *Kofoid, Charles A.—The Plankton of Echo River, Mammoth CEE. caldogamennbdcle on ood Ssh do septic daonboccbods deosbodporsccecr 113 Krauss, William C.—Some Medico-legal Aspects of Trauma in Re- lation to Diseased’ Celebral Arteries.. 2... 22.05. .0220.0220000 1 Lantern Slides of Microscopie Objects for Class Demonstra- tions, Methods of Producing Enlargements and, by John ANSTorIOn EMME occ sab oo Sod edd co doobddoDUOASouso OOO dO SasodOosKC 41 *Latham, V. A.—The Reaction of Diabetic Blood to Some of the PATVILID: Dy OSmarapsnvereee stoteislemeya\evctstorctets erelevatclaiatel ole sterstet ener ei etale'a!sPeialersvatele 3] *Library Expedients in Microscopy. Indexing, Cataloguing, Pre- paring and Arranging Literature and Slides, by R. H. Ward.. 127 *Limnobiology, A Plea for the Study of, by Henry B. Ward....... 201 iimavolosical ‘Commission, Report Of EAC. . eieieletete/aielcieleiee = ieee *Microscopy, Library Expedients in, by R. H. Ward............... *Minutesior the Ammittal Meetings <).:2)..0 «1. /ajois elesin sicielnio slelotestereiaieteiete Minutes) of the Amm mal’ Mee Gime ye s)- (5215 «1 che ot) ose.» were) «ole let ate lolchelateerenetale Mites, Description of a New Genus of North American Water, by OPT els PWV OCOD) vere sfeus oleioln ovedelers rei eloleielaeseleve) ote siciehiet=te iol teteie ean Modern Conception of the Structure and Classification of Des- mids, with a Revision of the Tribes and a Rearrangement of the North American Genera, by Charles E. Bessey.........«.. *Modern Conception of the Structure and Classification of Dia- toms, with a Revision of the Tribes and a Rearrangement of the North American Genera, by Charles E. Bessey............ *Moore, Veranus A.—An Incubator for Student Use.............. Mt. Prospect Laboratory of the Brooklyn Water Works, The Work of the, by George CG: Wihippleci ee. «6 sieiei<\«'-lelelelystohebreeietene *New Genera and Species of North American Hydrachnidae, by VO Er wEl VV OLCO GU ys icicislcieleintelemictercls olsintoteicvelale eterelstotei cies else netar nanan Nebraska, The Cladocera of, by Charles Fordyce................- PING CEOLO Osi laisie Sieletarsie in eiciele clei clelotale lls ciel e/ oie plafeie! siete clerala\elalchehetet-teet een ana INGLEROILIENA 55 boo dood. J6n DAG doo oo Ebon ane JOO oUOdDdboobor sooonscr New Avian Cestode, Metroliasthes lucida, by B. H. Ransom........ Notes on the Parasites of the Lake Fish. III. On the Structure of the Copulatory Organs in Microphallus nov. gen., by Henry Lae Eine So.qunccoG so LOU BBO Cogdedda to psoa oO do Obcealsolooo sor *Notices of Some Undescribed Infusoria, from the Infusorial Fauna of Louisiana, by J. C. Smith, New Orleans, La......... *Notogonia Ehrenbergii Perty, by J. C. Smith, New Orleans, La.. On the Distribution of Growths in Surface Water Supplies and on the Method of Collecting Samples for Examination, by Fred- EVICIE 55 | ELOMISS ee crspaleys sid cveveieter ais lelols si vere ecaeletoreteioieteletel tec kete etek ene IMMER EER eiaisniiere ore wiolic'e el euel eneiele. drills aid le se lbuel’e.-s) sy arinepa lebeiiev'e a teNey coe Ene nee EES POUR ea eke ese ce dere ie: alpha terete ve rels eiteiala, 6 lece wie Gls Vee eianeiar eave ty ate ener WAI Wie ccaisistals ie lolee oblate wicslemiieteleneteisicve) eye's aiejnin etelelet sit erste aleteiete tate nian Parker, Horatio N.—Some Advantages of Field-Work on Surface Water SUPPIIES oi icritousila etelstatsis wis o suslake cols tee cit (olalele tel en 1 Work of the Mt. Prospect Laboratory of the Brooklyn Water Works, by George C. Whipple... 2.2.2... -0.c0ss00+sseseeeescese 25 TRANSACTIONS OF THE American Microscopical Society ORGANIZED 1878 INCORPORATED 1891 EDITED BY THE SECRETARY Twenty-Fourth Annual Meeting HELD IN DENVER, COLORADO, AUGUST 29, 30, AND 31, 1901 VOLUME XXIII . LINCOLN, NEB.: JACOB NORTH & CO., PRINTERS 1902 OFFICERS FOR 1go1-1902 Ege dent: CHARLES Hg BESSEW 2\e,05 ieee aievaieinia coreaiels le tele neeetete Lincoln, Neb. Bae Presidents: Fo. A, UBRERGE, o).3) (HENRY BS WARD 372 oie) sya\c soln o s\a ny oe /siaieaielsl evsiave wis) eravalaens Lincoln, Neb. TD FEQSUPT ETS \]is Caro ME DES uiete rays ofes colo) ereysiaule)aie, ofa/elisielGlorelsy anereroe New Orleans, La. Crisp 100: NEA US PRE AGM 62:5 6/o. 0). 5b 0c:0-0,0 01 ale lod wet one OereRe Piitsburg, Pa. ELECTIVE MEMBERS OF THE EXECUTIVE COMMITTEE Ao MG NETO TMS Sass sliss dele revedebencterate bers piers lols Se wlae loi o} Glan e ERR ReR Senne Denver, Colo. Ni AS ACTA MAE Tey WAS Wate acelin robots ie labs eiciolie\ stare) a tee stave aye faletal eee et enenee Chicago, Ill. GNC. MVP BE Eee Sean cites tote ciel a iolal ast citelw a= 1b labial ola lel an tae New York City EX-OFFICIO MEMBERS OF EXECUTIVE COMMITTEF Past Presidents still retaining membership in the Society R. H. WARD, M.D., F.R.M.S., of Troy, N. Y., at Indianapolis, Ind., 1878, and at Buffalo, N. Y., 1879. H. L. SmirH, LL.D., of Geneva, N. Y., at Detroit, Mich., 1880, and at Cleveland, O., 1885. J. D. Hyatt, of New York City, at Columbus, O., 1881. ALBERT MCCALLA, Ph.D., of Fairfield, Ia., at Chicago, IIl., 1883. T. J. BURRILL, Ph.D., of Champaign, II1., at Chautauqua, N. Y., 1886. Go. E. FELL, M.D., F.R.M.S., of Buffalo, N. Y., at Detroit, Mich., 1890. FRANK L,. JAMES, Ph.D., M.D., of St. Louis, Mo., at Washington, D. C., 1891. MARSHALL D. EWELL, M.D., of Chicago, I11., at Rochester, N. Y., 1892. SIMON HENRY GAGE, B.S., of Ithaca, N. Y., at Ithaca, N. Y., 1895. A. CLIFFORD MERCER, M.D., F.R.M.S., of Syracuse, N. Y., at Pittsburg, Pa., 1896. W. C. Krauss, M.D., of Buffalo, N. Y., at Columbus, O., 1899. A. M. BLEILE, M.D., of Columbus, O., at New York City, 1900. C. H. EIGENMANN, Ph.D., of Bloomington, Ind., at Denver, Colo., 1901. The Society does not hold itself responsible for the opinions expressed by members in its published Proceedings unless endorsed by a special vote. TABLE OF CONTENTS FOR VOLUME XXIII The Annual Address of the President, The Solution of the Eel Ques- tion, by Carl H. Eigenmann, with Plates I toIV.................- 5 The Debt of American Microscopy to Spencer and Tolles, by William C. Krauss, B.S., M.D., Buffalo, N. Y., with Plates V to IX........ 19 A New Species of Crenothrix (C. manganifera), by D. D. Jackson, C2 iM oo Ee Orne t si EME He Pen tne TN em eat Nee LA SRS AMR le eA 31 Notes on Colorado Entomostraca, by Arthur E. Beardsley............. 41 Notes on Colorado Protozoa with descriptions of new species, by Arthur Ey, Bearisley,, with Plate: ol, 6 ia sicias etal: sis ais'ein's wince SAS weeislabeaeys 49 The Plankton of Lake Maxinkuckee, Indiana, by Chancey Juday...... 61 The Morphogenesis of the Stigmata and Stomata occurring in Perito- neal and Vascular Endothelium, by Arthur E. Hertzler, A.M., ED with, Plates SebE ad DET oi)! yolaiwin's,aisisin wish ow u's h'loioie 4/siahejajele 63 A Contribution to the Subterranean Fauna of Texas, by Carl Jost Ul- CHER WILE ates XC VAR LO PNW Denso tesrsie ieieycig) sleiave obs efe else) Solo eter eee 83 On the Amount of Oxygen and Carbonic Acid Dissolved in Nitural Waters and the Effect of these Gases upon the Occurrence of Microscopic Organisms, by George C. Whipple and Horatio N. Parker, with Plates XIX to XXII The Structure and Classification of the Conjugatae with a revision of the families and a rearrangement of the North American genera, ye enles ie messey. PMD.) nic cic'as a cle daisielera «'0!s,s@. s/s s1a\sinfelatnivieters 145 On Aymenolepis carioca (Magalhaes) and Aymenolepis megalops (Nitzsch) with remarks on the classification of the group, by B. H. Ransome wither lates xl CO oN Vere sareicls sate ale/elaijaieleiclierciers! s olatelsievcls 151 Studies on the Genus Cittotaenia, by Rufus Ashley Lyman, with Plates PRONG ATC AT PRONG Olea yet ciey alot at ntepey ayane aval lalieieye) ota stale) sVatalalors\ einiete/atelie) orale 173 Some Points in the Structure of the Acanthocephala, by H. W. Gray- UO Symi Ure OO 00 RR pcm ob obo DUE GOOD COnoDUDSCOOeS 191 The North American Species of Curvipes, by Robert H. Wolcott, with PRES Re NO EG ONO DE 5 sic sicl dala Satay at araioreialaieialarelevarn ols irae ctainle miami ta 201 Ae Newstigars. by) Mi Js Felrod 6 .iec:06.- «n<\o/s0e winercla’e « enia(ainlelm eae ate 257 Modification of some Standard Apparatus to facilitate the Work of the Histologic and Embryologic Laboratory, by Simon Henry Gage, With Plate XEXPRD Ve yt 1al ate ere a (ainjele auaile's atnl's late repemetetae mialenst aia a al 259 Laboratory Photographic Apparatus, by Simon Henry Gage, with Plates SORRY) ta URN ERO foo ata Sibiu spaiarevaras shave w/eiejelnletsabtenctenetaiatametal steve ate) s 263 CONTENTS List of Subscribers to the Spencer-Tolles Fund..............cceeeeeees 267 Wecrology, E. W. Claypole, with Plate: 3... ....00% suis ce welsele eee 269 Minutes of the Annnal Meeting. oo. .....6c5 ois oie sen ele se 275 Treasurer’s Reports.) 2-4 sels Utica «bes eeolnis: tele late siala’s (Geis erele ieee 281 Custodian’s Report, Spencer-Tolles Fund, 02. 600532 0:05.00 cenit 282 Constitution cies 2) acre ies erste sale mie evar eto teveralele ielehel cls etoleitelevcposlay enn eee 283 By Baws wosvzeicle-cteciniess Ge aikle bits eve, Sisvaie otk) beim ciw's ntwieieie ao le ai teayoielo ye) seek 284 TASH Or Mler ETS ies =» cictasin'e.cytis ais inte, 5:05 6 wlelaio/s bi s\e wiasal © Nebgeie wine ee 287 Tjist Of Subscribers. o 2. cence eesiss viele calele wisisw e's nce sens seine 294 Advertisements achiev cite cocg wm cldennre co cleteidle alee slacclelelavlotelcveteteleters strates oe TRANSACTIONS OF The American Microscopical Society TWENTY-FOURTH ANNUAL MEETING, HELD AT DENVER, COLORADO, AUGUST 29, 30, AND 31, I9OI THE ANNUAL ADDRESS OF THE PRESIDENT THE SOLUTION OF THE EEL QUESTION By CARL H. EIGENMANN WITH FOUR PLATES Eels are abundant in nearly all the rivers of Europe and the At- lantic slope of the United States. No eels with ripe eggs and no young eels less than three inches long have ever been found in any of these streams. The question naturally arose, How, where, and when do eels reproduce? This, the original eel question, was modi- fied when it became known that the adult eels migrate to the ocean in a sexually immature condition, and young eels enter the mouths of streams and become distributed throughout their length. The eel question was seriously considered by Aristotle several centuries B. C., and in 1880 A. D. Jacoby wrote, “To a person not acquainted with the circumstances of the case it must seem aston- ishing, and it certainly is somewhat humiliating to men of science, that a fish which is commoner in many parts of the world than any other fish, the herring perhaps excepted, which is daily seen in the market and on the table, has been able, in spite of the powerful aid of modern science, to shroud the manner of its propagation, its birth, and its death in darkness, which even to the present day has not been completely dispelled.” 6 CARL H. EIGENMANN Many fishes migrate, preliminary to spawning, to regions other than those in which they usually live. It is well known that the salmon of the Pacific slope spend the greater part of their lives in the ocean. When they become sexually mature they run up some stream to deposit their spawn near the head-waters. They may ascend for a thousand miles or more, through rapids and over falls ten feet high and more, to an elevation of many thousand feet.* After they enter fresh water they rarely take food. The exhaus- tion incident to their long journey and the wounds they receive on the spawning beds in cleaning the gravel or in fighting with their rivals prove fatal to all of them. They all die shortly after spawn- ing. The young salmon remain for a variable time where they are born, then descend the streams to the ocean, where they remain till they in turn are sexually mature. The history of the fishes, as long as they are in the streams, can be followed without any great diffi- culty, but their doings after they enter the less confined regions of salt water are not easy to trace. The salmon are not the only migratory anadromous fishes. Vari- ous other species regularly ascend streams to spawn. Others show a tendency to enter fresh water, or at least brackish water, during the spawning season.?, Even many fresh-water fishes migrate up- stream before the spawning season. While many species of fishes have the habit of entering fresh water when they approach ripeness, the eel alone has the reverse habit of taking to salt water when the reproductive period ap- proaches. It has been well known for many years that during winter and early spring the young of the eel enter the mouths of streams in enormous numbers. (Redi records the entrance of young eels into the Arno in 1667, and that at Pisa three million pounds of young eels 30-120 mm. were taken in five hours.) They find their way for hundreds of miles from the ocean. “Young individuals three to five inches long ascend rivers in incredible numbers, over- coming all obstacles, ascending vertical walls or flood gates, enter- ing every large and swollen tributary, and making their way even over terra firma to waters shut off from all communications with 1For instance, the elevation of Alturas Lake, one of the spawning grounds of the Chinook and Qinnat Salmon, is 7,335 feet. 2 During an unusual freshet at San Diego, Cal., in 1889, large numbers of ripe Cynoscion nobile ascended the temporary fresh-water streams. Its relative in the Atlantic coast, Cyno- Scion regale, spawns in the brackish water at the mouths of small streams. : THE SOLUTION OF THE EEL QUESTION 7A rivers.”° While in the fresh water they feed on everything eatable. When they approach their full size, in about four years, they de- scend (in autumn) the streams to the ocean, where they are lost sight of beyond a distance of a mile from shore. After these facts were established the eel question became: What do the adult eels do after they enter salt water, when and where do they produce their young, and where do they die? In how far these questions have been solved and the method of their solution are the topic of the present paper. The solution has been approached along at least three distinct paths. The search for the reproductive organs. Three different theories have been held as to the reproduction of the eel. Aristotle supposed that “the eel is neither male nor female and is procreated from nothing; . . . no other animal pro- duces young without eggs, but no eel has ever been found to con- tain eggs.” “They are produced from the so-called ‘bowels of the earth’ [earthworms] which are spontaneously produced from mud and moist soil.’”’ Aristotle contended against another idea preva- lent to some extent even in his day—that the eel produces living young—stating that the supposed young eels found in the eel are intestinal worms. The naturalists of the Middle Ages generally believed that the eel produces living young. Associated with the idea of Aristotle that eels are spontaneously created through the mediation of earthworms are the notions that the eel is produced by other fishes or even other animals, and in vari- ous regions different creatures serve the fisherman as the mother of the eel. A blenny in the north of Germany, a mullet in some parts of the Mediterranean, and a beetle in Sicily, and horse hairs in many parts of the world have been looked upon as the progenitor of the eel. Interest in the reproduction of the eel has always been kept up. When Schulze died he is said to have expressed the consolation that all the important questions except the eel question had now been settled. During the sixteenth century, according to Jacoby, the discus- 7) 8Mtr. Gordon Land, formerly Fish Commissioner of Colorado, told me of the presence of eels in streams of the Los Ojos ranch, Colorado, a distance of about 1,500 miles from the Gulf of Mexico, and at an elevation of about 7,200 feet. 8 CARL H. EIGENMANN sion of the eel question was limited to the young ones reported to have been found in them. In 1707 Vallisneri published an account of the supposed ovary of an eel sent him by a friend from Co- macchio. This eel was distended and contained a body resembling an ovary containing eggs. During the middle of the eighteenth century the fishermen, desirous of gaining the liberal rewards offered for ripe eels, began the perpetration of frauds on the ambitious naturalists, which has continued to our own day. Professor Moli- nelli received an eel previously stuffed with the eggs of another fish. In 1777 a council of naturalists, containing among others Mondini and Galvani, sat about another apparently ripe eel caught near Comacchio. Mondini reported upon this eel and showed that the structure described by Vallisneri seventy years before as the ovary was only the diseased air bladder, and himself described the true ovary. The ovary was independently discovered by O. F. Muller a few years later. Spallanzani, after a study of the eel question at Comacchio with purely negative results, rejected the discovery of Mondini, and the latter’s observations were not confirmed till 1824, when Rathke independently described the ovary of the eel, and later, in 1850, when the same author described a female with eggs 0.1 mm. in diameter filling the whole abdominal cavity. After Rathke’s description of the ovary of the eel a number of authors took up the hunt for the male reproductive organs. Fora time the fatty bodies found in female eels were taken to be the male reproductive glands, and eels were supposed to be hermaphro- dite. A new path for research was pointed out when Darwin in his Descent of Man quoted Guenther to the effect that the males of fishes are smaller than the females. Syrski in 1874 published the first account of the male repro- ductive gland of the eel. While the discoveries of Mondini, Rathke, and Syrski demonstrated the presence in eels of reproduc- tive organs such as are found in other fishes, we were brought no nearer the solution as to the time and place of reproduction or of the reproductive habits of the eel. A reward was consequently offered by the German Fishery Association for any eels sufficiently well developed sexually to advance the knowledge of the repro- ductive history of the eel. The only result seems to have been jocose remarks in the funny papers and a continuation of the at- tempts of the perpetration of frauds on the parts of fishermen. THE SOLUTION OF THE EEL QUESTION 9 In 1877 Jacoby visited Comacchio to learn what he could con- cerning the modifications of eels after they had entered the sea. He found that of eels taken indiscriminately 5 per cent were males, whereas 20 per cent of those less than 45 cm. long were males. He also found that males took part in the fall migration to the sea. He did not succeed in finding eels as much as a mile from shore, and none of those from shallow water near shore showed reproductive organs more advanced than those from fresh water. He concluded that eels must mature in deep water in the ocean and that they die after the spawning season. Some side lights have been thrown on this part of the eel ques- tion by observations on the marine or conger-eel. !t has been found that some weeks before it reaches ripeness the conger-eel ceases to eat. The eggs and sperm reach maturity im individuals kept in confinement, but they can not be liberated under the condi- tions obtaining in confinement and the fish die; in some cases it has happened that the fish burst as the result of the accumulation of ripe eggs which could not be liberated. The feeding habit of the conger-eel thus agrees with that of the Pacific salmon, and it is very probable that the fresh-water eel also stops eating some time before it reaches ripeness. The stomachs of eels migrating to the sea were always found empty by Jacoby. The conclusions from the series of observations recorded above were that eels have reproductive organs like other fishes, but that they do not reach maturity in fresh water, and that for this reason the difference between the sexes of eels while they are in fresh water are inconspicuous; also that the male eels are, on an average, much smaller than female eels. ‘The inferences were that eels re- produce as other fishes do, and that reproduction takes place in deep water after the period of maturation during which no food is taken. The discovery of Leptocephali. While the present phase of the eel question was being approached by the study of the reproductive organs and habits just described, it was being approached trom two other directions. Over two hundred years ago Scopoli (1777) discovered a pecu- liar, transparent, flat, ribbon-shaped fish with minute head and tail. Others were discovered later, and up to 1895 twenty-five or more species of these fishes, called Leptocephali, were described. The Io CARL H. EIGENMANN extreme transparency of these Leptocephali is strikingly shown by Leptocephalus diptychus (pl. I, figs. 1, 2), a new one described dur- ing the past winter (Science, XIII, 828. 1901). This Leptoceph- alus differs from all others in having a series of seven conspicu- ous black spots along the middle of the sides. On close inspection it was found that three of these spots are on one side and four on the other, that the spots of opposite sides alternate with each other, and that the extreme transparency of the larva results in the blend- ing of the two alternating rows of opposite sides into a single series of spots. Still other transparent, more cylindrical fishes, slightly more like eels, were described under the name Helmichthys. The longest of the Leptocephali captured measured 250 mm. They were for a time considered to be a distinct group of fishes. In 1861 Carus studied these forms and came to the conclusion that they are but early stages of other fishes. In 1864 Gill definitely recognized one of the Leptocephali (L. morissii) as the young of the conger-eel, and the others were supposed to be the larvae of various eels. The ques- tion now arose whether the Leptocephali were normal stages in the development of eels or “whether they are individuals arrested in their development at a very early period of their life, yet continue to grow to a certain size, without corresponding development of their internal organs, and perishing without having attained the character of the perfect animal.’ From the fact that the longest young eels were shorter than the longest Leptocephali Guenther favored the idea that Leptocephali are abnormal larvae. In 1886 Delage (C. Rend., CIII, 690.) published his observations on the actual transformation of a Leptocephalus into the conger-eel, and thus demonstrated the fact that the Leptocephali are normal larvae. The discovery of ripe eel eggs. The third and last path to the present phase of the eel question was discovered by Raffaele. The Italians being most favorably located have done most toward the solution of the eel problems. In 1888 Raffaele described five species of pelagic eggs from the Bay of Naples which, on account of the larvae into which they developed, he referred to various unknown species of eels. He was able to 4Other fishes have Leptocephalus-like larvae differing from those of the eel largely in the fins and tail. In 1889 Professor Gilbert showed me complete series of stages, from a long slender band-shaped Leptocephalus to a much shorter individual of A/bula vulpes. They were taken by him in a shore seine on the coast of Lower California. THE SOLUTION OF THE EEL QUESTION II keep them for fourteen to fifteen days only and was therefore un- able to determine to what particular species of eels they belonged. The eggs had al! the characteristics and habits of pelagic eggs.° Grassi later found the same eggs at Naples. The eggs described by Raffaele have certain characters in com- mon: They are much larger than average sized pelagic eggs; they have a very large perivitelline space; their yolk is vesicular; they differ from each other in size and the presence or absence of an oil sphere. One of the eggs, his No. 10, has a diameter of 2.7 mm. and is without an oil sphere. It develops into a larva with 44 (45) abdominal segments. All of them were taken between August and November, being more abundant in September. When the larva is five or six days old it is slender and elongated with a greatly compressed body, very transparent, and with little pigment. The vitellus is very elongated and diminishes from in front backwards. The intestine ends in the ventral fin fold a short distance from the body in a small accumulation of cells. The noto- chord is formed of a single series of segments. During the second day after hatching the mouth opens. The teeth develop rapidly. Three pairs are developed in the upper jaw. This dentition is abso- lutely exceptional among fishes. Contemporaneously with the de- velopment of the mouth the choroidal pigment and five or six black pigmented spots form along the body. No noteworthy changes take place between the fourth and fifth day after hatching. Beyond this time he was unable to rear the eggs. The identification of the egg and larva of the European eel. The capping stone for the triple arch constructed by the anato- mists, descriptive zoologists, and embryologists respectively was brought by Grassi in 1897. Grassi begins the English abstract of his work as follows: “Four years of continuous researches made by me in collaboration with my pupil, Dr. Colandruccio, have been crowned at last by a success beyond my expectation, that is to say, have enabled me to dispel in the most important points the great mystery which has hitherto surrounded the reproduction and the development of the commoneel. . . . The most salient fact dis- covered by me is that a fish, which hitherto was known as Leptoceph- 5Raffaele. Le uova galleggianti e le larve dei Teleostei nel golfo di Napoli., Mith. aus der Zool. Station zu Neapel, VIII, pp. 1-84, tav. 1-5. € None of the eggs taken from the fresh-water eel exceed 0.27 mm. diameter. I2 CARL H. EIGENMANN alus brevirostris, is the larva of the Anguilla vulgaris.” He was able to follow a Leptocephalus through its metamorphosis into the common eel. He supposed that normally these processes go on at a depth of at least five hundred meters. His work was carried on on the coast of Sicily, where strong tidal currents cause the dis- placement of the water in the narrow Strait of Messina. As the result of these currents all stages in the development of the Murae- noids are sometimes met with in the surface water. They are also abundant in the stomach of the sun-fish, Mola. He found that the male eels may ripen in shallow water and migrate when ripe to deeper water. Some eels approaching ripe- ness were found in the sewers of Rome. The ripe male eel has taken on a silvery color and its eyes are very much larger than those of the river eels.’ The females never ripen in shallow water. The eggs he sup- poses are laid at a great depth and remain suspended at a great depth, only occasionally reaching the surface. Instead of being small, as had been supposed, they are really much above the average of pelagic eggs in size. From the eggs come prae-larvae which de- velop into the regular larvae or Leptocephali (fig. 4) with the anus and urinary organs near the tip of the tail. The larvae are metamor- phosed into what he calls hemilarvae with the two openings mov- ing toward the permanent position. By further changes, which include a considerable reduction in length, the hemilarva assumes the shape of the adult. Both the larva and hemilarva are longer than the young eel arising from them, there being a diminution of 4 cm. in length during the transformation. He found that the caudal fin of the Leptocephalus always resembled the caudal fin of the adult eel into which it developed and that the number of seg- ments is also a constant character. He identified Raffaele’s egg No. 10, without oil sphere and with a diameter of 2.7 mm., to be that of the common eel. The discovery of the Leptocephalus of the American eel. During the past winter while describing the Leptocephali belong- ing to the United States National Museum one of my students, Mr. Clarence Kennedy, and myself discovered two specimens taken on 7Bean in the Nineteenth Report of the Commission of Fisheries of New York, p. 28), de- scribed five male eels with well-developed reproductive organs which were probably ordinary eels in their nuptial dress. He describes them as having “large eyes, short snout, and long pectoral fins, as compared with the conimon form, silvery gray above with a clear satiny white abdomen separated from the color above by the lateral line.’’ THE SOLUTION OF THE EEL QUESTION 13 the surface of the ocean off New York that in shape, color, etc., very greatly resembled the Leptocephalus of the European eel. When we found that the American eel had fewer vertebrae than the European eel, and that the larvae under consideration differed from the larvae of the European eel in just those characters in which the American eel differs from the European, we felt certain that we had found the larva of the American eel (fig. 3). The development of the conger-eel. During August of 1900 I was fortunate enough to secure the eggs of an eel, very probably that of the conger-eel. They were taken by Dr. Porter E. Sargent on the U. S. F. C. vessel Grampus from the surface of the Gulf Stream. They are the first eel eggs that have been secured outside Italian waters. Since in their de- velopment they greatly resemble the development of the fresh-water eel their history may be added. The eggs secured by Dr. Sargent measure 2.4-2.75 mm. from membrane to membrane. The yolk, as in the eggs described by Raffaele, is made up of transparent spheres not unlike those of the eggs of certain clupeoids. There are present from one to six light yellow oil spheres of variable size. If more than one are present, then one is always much larger than any of the others. The yolk measures I.75-2 mm. Some of the young were found to be hatched on the morning of August 3. Since many of these de- veloped gaping jaws, and some others, which did not hatch till several days later, developed normally, it is possible that the early hatching was not normal. Raffaele’s eggs hatched in five or six days. He was able to keep them four or five days after hatching. For some time after hatching the larvae floated with their heads up- ward—the probable result of the location of the oil spheres. On August 6 they had assumed a normal horizontal position and the characteristic eel-like progression, but the pectorals were not yet used in swimming. Later they were seen eeling their way through the water, not infrequently nosing about the bottom and voraciously seizing anything that came in their way. The characteristic feature of the eggs at the time I began to ob- serve them, August I, was the shape of the yolk. The bulk of this occupied the usual position, but a narrow stalk extended back below the alimentary canal. The oil sphere or spheres occupied the ex- I4 CARL H. EIGENMANN treme anterior part of the yolk® (fig. 5). The further history of the yolk in this species is unique among fishes and not sufficiently em- phasized by Raffaele. In fig. 6 it is seen that the yolk is no longer rounded anteriorly, but that it ends in a marked protuberance and that the oil sphere lies in this. The general mass of the yolk still retains the original shape and distribution. The anterior protuber- ance now becomes longer and at the same-time narrower, so that the oil sphere loses its rotundity and becomes elongate (fig. 7). At the same time the general mass of the yolk diminishes rapidly in the yolk sac, while in the elongated pouch along the ventral side of the alimentary canal no diminution is evident. On the contrary, there is an apparent increase; the entire yolk sac becomes notably longer with the increase in the length of the body. Very soon (fig. 7) the oil sphere, much elongated, with a small surrounding mass, is all that remains as a spindle-shaped figure in the yolk sac. The yolk sac does not at once lose its shape and bulk, but serves as an unusually large pericardial chamber which is equaled only in the practically yolkless Cymatogaster. On August 5 the yolk along the alimentary canal had suffered little diminution, and its outlines were quite regular (fig. 8). On August 6 this part of the yolk had become constricted in places, the outlines being less regular (fig. 9). The yolk had become yellowish in color and more fluid than vesicu- lar. On the following day (fig. 10) the constriction had deepened, and on August 11 the remains of the yolk were located in a series of minute globules more or less widely separated from each other (fig. 11). Long before this condition was reached, about the 8th of August, the larvae were taking food. The number of segments developed in front of the anus differs slightly, ranging from sixty-five to seventy-one. The number be- yond this point could not be determined exactly. The notochord consists in its anterior fourth of single segments. In its middle region the segments do not extend through its entire thickness, but in the tail it is again formed of single segments. The lines separat- ing these are so much more conspicuous than the lines separating successive myotomes that it is impossible to make out the latter in the thin transparent tail. _ 8All drawings were made from living specimens, or such as had just been killed by form- rE es the drawings, a/, alimentary canal; /v, fourth ventricle; »/4, yolk; J, liver; hk, heart; 0, oil spheres. THE SOLUTION OF THE EEL QUESTION 15 Color is late in making its appearance. It is first evident at the end of the tail. At 6:00 p.m. on August 5 some of the larvae had the following six spots above the alimentary canal and along the lower margin of the myotomes of the tail: (1) About the middle of the yolk, (2) halfway between this and the end of the yolk, (3) at the end of the yolk, (4) in front of the anus, (5) some distance behind the anus, (6) about the tip of the tail. Additional spots are added between these already formed. The relative and actual size of the spots differ greatly, but the number is the same in different specimens of the same age. In the oldest larvae the spots repre- sented in fig. 11 were developed. Aside from those along the lower part of the sides there were a few cells on the upper jaw, and the scattered cells seen near the tip of the lower jaw as early as August 7 (fig. 10) have developed into a well-marked spot. The character of the pigment about the tail is also noteworthy. In the last stage figured the processes of the cells show a tendency to lie parallel to the embryonic fin rays. Pigment is formed in the eye with its earliest appearance on the body. No color, aside from the black pigment spots and the yellowish yolk, is seen anywhere about the larva during the time the larvae were under observation. The fin fold is well developed, reaching from the nape around the tail to the yolk sac. It is much wider along the back and in the region of the vent than about the tip of the tail or the ventral line of anterior abdomen. No rays had appeared in the oldest larvae observed except about the tail, where there appears a distinct radiation. The enormous development of the posterior half of the fourth ventricle is similar to the condition figured by Raffaele. In all but the last stage figured this part of the fourth ventricle is a large thin- roofed vesicle, separated from the fin fold in the earlier stages by a distinct notch. The auditory capsules are conspicuous, and, viewed from above, are seen to protrude from the sides of the head. The alimentary canal is marked (1) by large fang-like teeth, (2) the early vesicular development of the liver, (3) and the posi- tion of the anus near the body and remote from the margin of the ventral fin fold. As soon as the mouth is open, about the fourth or fifth day from the beginning of development, the margins of the jaws are seen to be marked by small protuberances. These are the swellings within which the teeth are developing. In the upper jaw 16 CARL H. EIGENMANN four pairs of teeth are developed, graded from in front back, the anterior ones being comparatively enormous fangs. In the lower jaw four pairs are also developed. These are more uniform in size, but with the second one larger than the rest. In the oldest indi- vidual there were five pairs of teeth in the lower jaw. I am unable to say whether this was a normal condition. The teeth of the upper jaw close over those of the lower jaw (fig. 12). The oesophageal pouch (liver) of Raffaele has already been men- tioned. Even before hatching it is a conspicuous pouch behind the heart. Later, when the anterior yolk has been largely consumed and is separated from the posterior yolk by a constriction, the vesicular structure becomes converted into a lobulated organ about this constriction. Conclusions. We now know (1) that eels, both male and female, migrate to the ocean during October to January; (2) that these eels probably de- posit the eggs that are found on the surface during the following August to January; (3) that the eels do not ripen in shallow water, but the female, according to Grassi, at a depth of five hundred meters; (4) that the eggs of the eels float, according to Grassi, at a great depth; according to Raffaele and Eigenmann at the surface; (5) the development of some eels for the first fifteen days and that the resulting creature is different both from the adult eel into whicli it will develop and from the larva of the eel; (6) the Leptocephalus of the eel and the process of its metamorphosis through a Hemich- thys stage into the young eel as it is found entering the streams; (7) the young eels enter the streams during spring about two years after their parents have entered the sea. We do not know the history of the larva from an age of fifteen days till they reach the Leptocephalus stage. We do not know for a certainty that the egg and early stages of development of the common eel has been secured, although it is very probable that Raffaele’s No. 10 is the egg of the common European eel. We are not certain that the egg is normally or only occasionally pelagic. We do not know the normal habits of the Leptocephalus. We have not yet secured a female eel with eggs larger than 0.27 mm. _Inas- much as the mature egg probably reaches a diameter of 2.7 mm., the largest ovarian eggs found must increase a thousand-fold before maturity is reached. THE SOLUTION OF THE EEL QUESTION 17 The question whether or not the eel ever breeds in fresh water has been answered in the affirmative by several observers. There is nothing that would indicate the inherent impossibility of eels becoming land-locked and breeding in fresh water. The evidence is, however, so far inconclusive. No one has yet taken eel eggs or larval eels or younger eels than those that ordinarily ascend streams from the ocean in any fresh water. The statement that they must breed, because we know of no other way in which the supply of eels is being maintained in land-locked basins is not conclusive evidence that they do breed in these basins. Feddersen® states that in the north the eels have in places be- come strictly fresh-water species which can be distinguished from the migratory eels by definite characters. Imhof? concluded that eels breed in fresh water on the following evidence: In 1882, 1886, and 1887 a large number of eels were placed in the Caumasee (Canton Graubtinden). After 1887 no ad- ditions were made. In June, 1895, a small male eel 47 cm. long was taken among other eels. From the impossibility that eels should have arrived from the ocean by migration, since the Caumasee be- longs to an isolated water-basin, the presence of this small eel was supposed to demonstrate the fact that the eels had propagated in this lake. The evidence does not seem to me to be conclusive. Male eels are much smaller than female eels, whereas nothing is known to the contrary that the time required by the small male to reach its full size is not as great as the time required for the female to reach her full size. If the time required be the same, then the finding of a male eel 47 cm. long is no more evidence of recent breeding in this lake than the presence of female eels 11%4 m. long. The ques- tion whether or not the eel ever breeds in fresh water may be con- sidered undecided. 9 ‘Ueber das Laichen des Aales im Siisswasser,” Zeztschrift f. Fischeret u. deren Hilfwiss- enschajten, pp. 156-67. 1895. 10 Bioiogisches Centralblatt, XVI, p. 431. 1896. 8 CARL H. EIGENMANN EXPLANATION OF PLATES Piate I Fig. 1. Leptocephalus diptychus, a and 6, enlarged views of the head and tail. Fig. 2. Leptocephalus diptychus, an older larva; a and 6, enlarged views of head and tail. Fig. 3. Leptoczphalus Grassti, the larva of the American eel. Plate I Fig. 4. The metamorphosis of the European eel after drawings by Grassi. Figs. 5 to 12 refer to the development of the Conger-eel. Fig. 5. Outline of embryo showing position in membrane and shape of the yolk. Aug. 1. Fig. 6. Embryo freed from its membrane, showing the beginning of the constriction of the yolk at its anterior end. Aug. 1. Plate Il Fig. 7. Head and anterior part of body showing the continued reduction of the yolk and the very large fourth ventricle. Aug. 1. Fig. 8. A larva on Aug. 5. Fig. 9. A larva on Aug. 6. The mouth probably slightly abnormal. Fig. 10. A larva on Aug. 7. Plate IV Fig. 11. A larva of Aug. 14. The fin fold of this larva is probably repre- sented as too low. Fig, 12. Dentition of a larva of Aug. 14. Fig. 13. The full-grown Leptocephalus Morisstt, the larva of the Conger- eel; a and 4, enlarged views of the head and tail. Fig. 14. An older Leptocephalus Morissit undergoing its metamorphosis; a and 6, enlarged views of the head and tail. PLATE f Pagid Sy) ADDI») DD YEE SSyy>dd>d >a DP »y)) YY) / Stk PIATE It UU J PLATE Ill PEELE EEE ARICA _imm., Ze RAN PLATE, IV. £9 13 >) «ld TU RY AWAY «KK CS SSS \ 2 SOO YEA HOHHS ? VEIL — a Se a KG \ THE DEBT OF AMERICAN MICROSCOPY TO SPENCER AND TOLLES By WILLIAM C. KRAUSS, B.S., M.D., BUFFALO, N. Y. PAST PRESIDENT OF THE A. M. S. WITH FIVE PLATES There recently died in Buffalo the last of a distinguished trio of lensmakers who, by their ingenuity and inventiveness, added a bril- liant chapter to the history of American genius. Charles A. Spencer, Robert B. Tolles, and Herbert R. Spencer were artists rather than artisans in their chosen field of applied optics, and they left their impress deeply engraved upon the history of their time. Although their obituaries have appeared in the Transactions’ of this Society at the proper time, nevertheless it will not be uninterest- ing or tedious to review their work or recall their efforts in behalf of improved scientific apparatus. Records show that up to 1840 little if any use was made of the compound microscope in America, and no instrument maker had appeared who could supply an instru- ment of any kind. Thus it was when in this year the United States exploring expedition to the South Seas under Commodore Wilkes was fitting out, no instrument could be furnished the expedition by any of the makers of scientific or philosophical instruments in America. In this dilemma a private individual was applied to, and an instrument was finally loaned from Dr. Paul Goddard, of Phila- delphia. It was a French microscope of inferior make, but the best obtainable at that time. Since then the instrument has come into general use, and in certain departments of the manufacture of micro- scopes this country has become preeminent. Scarcely had the Eng- lish microscope makers published those inventions and discoveries which rendered achromatic microscopes really possible, and ele- 1Memoir of Charles A. Spencer by Hamilton L. Smith, LL.D. Transactions American Sociely of Microscoptsts, 1882, p. 49. Memoir of Robert B. Tolles by George E. Blackham, M.D. Tyvansactions American Soctety of Microscopists, 1884, p. 41. 20 WILLIAM C. KRAUSS vated the instrument from the position of a mere scientific plaything to that of an instrument calculated for the most accurate investiga- tions, before the elder Spencer succeeded in producing lenses which at once took a front rank among the art productions of the world. To-day we step in the mad whirl of our busy lives to pay homage to Charles A. Spencer and his two famous pupils, Robert B. Tolles and Herbert R. Spencer. At the Seventh Annual Meeting of this Society, held at Rochester, N. Y., August 19 to 22, 1884, a memoir of Robert B. Tolles, but recently deceased, was read by Dr. George E. Blackham, of Dun- kirk, N. Y. In the remarks following by different members of the Society, a resolution was offered by W. H. Breasley, of Detroit, that W. A. Rogers be requested to prepare a subscription paper and to receive subscriptions for a monument to Robert B. Tolles. As a substitute motion Professor T. J. Burrill, of Champaign, IIL, offered the following, which was seconded and adopted: Resolved, That W. A. Rogers, H. J. Detmers, and George E. Blackham be made a committee to report upon proper action, on the part of the Society, in memory of Robert B. Tolles. Mr. Breasley then offered the following resolution: Resolved, That the same committee be asked to consider and report upon a suitable memorial for Charles A. Spencer, which was also seconded and unanimously carried. Thus the ground was broken for the foundation of a monument to Spencer and Tolles, which was to be as durable and ineffaceable as a granite shaft, but, unlike the stone, it was to shed warmth and awaken a quickening in the minds of all those fortunate enough to come within its shadows. At the next meeting of the Society, held at Cleveland, O., August 18 to 21, 1885, Dr. George E. Fell, of Buffalo, N. Y., reported the condition of the Spencer-Tolles Memorial Fund as follows: To the Officers and Members of the American Society of Microscopists: In accordance with the resolutions on a Spencer and Tolles Memorial Fund, the following report is presented: The first cash subscription to this fund was made by the Royal Microscopical Society, December 17, 1884. Since that time the subscriptions have come in so slowly that this report will present but a meager list of subscribers, and, in view of the unanimous adoption of the reso- lution establishing the fund, not nearly so large a list as should have been ex- pected. Prof. Wm. A. Rogers, with his characteristic action in furthering any DEBT OF MICROSCOPY TO SPENCER AND TOLLES 21 of the projects of the Society, has offered to subscribe $25 and guarantees $15 ad- ditional, contingent, however, upon a concerted action of the Society towards the increase of the fund. He suggests that the income of the fund be awarded in prizes for specific original research. The subscriptions to the fund are given below: Royal Microscopical Sociely. < ci MeSOx \ Kaen nvemas 8.0 16.0 1.0 Wa Gli ir Sens 53 7.8 {es Undetermined..... 35) Bi" J, Total mineral ...... 67.1 52.2 133.9 Total organic....... 22.4 5.8 21.1 Total mineral and CRG RANINO iro siosess! W/e acd 89.5 58.0 155.0 A NEW SPECIES OF CRENOTHRIX 37 It will be seen that waters which produce Crenothrix kuhniana contain a predominating amount of iron, that waters which produce Crenothrix manganifera contain a predominating amount of manga- nese, and that waters which produce Crenothrix ochracea contain a predominating amount of aluminum. It is probable also that this aluminum must be present in the form of sulfate or in combination with carbonic acid and organic matter. Sulfate of aluminum in quantity has been found in waters near these wells and may occur as such in the waters affected. Crenothrix kuéhniana was grown artificially in an atmosphere of air and carbonic acid. The cultures were in a mixture of agar and gelatine containing ferrous and manganous sulfates, and were kept in the dark at room temperature. The growth was not very luxuriant, but took place in sufficient amount to show that the or- ganism in growing threw down but little manganese, but selected the iron to precipitate. This was also proved in stagnant solutions containing salts of manganese and iron. In the same way cultures of Crenothrix manganifera were grown in solutions containing iron and manganese, and although the growths were but slight they showed the dark color of the manga- nese predominating in the precipitate. It is highly probable that Crenothrix ochracea would in the same way always select aluminum in predominating amount to pre- cipitate, as out of many hundreds of waters containing Crenothrix which have been examined there has been no case where this species has thrown down a predominating amount of iron or manganese, and in no case has Crenothrix kihniana been known to throw down a predominating amount of aluminum. In other words, the species always precipitates in predominating amount the special hydroxid ascribed to it. Crenothrix kuihniana was also grown on gelatin agar in an atmos- phere of hydrogen. The growth was normal in measurement and developed well-marked filaments; but although the medium con- tained ferrous sulfate, none of the iron was thrown down. This is because there was no opportunity for oxidation. We can conclude that Crenothrix kihniana is very common, be- cause iron in quantity is a common constituent of water; that the species C. ochracea is rather uncommon because aluminum sul- fate or aluminum combined with carbonic acid is less common in 3 38 D. D. JACKSON water; that the species C. manganifera is very rare because manga- nese seldom occurs in sufficient quantity in water to produce a noticeable growth of the organism; that the growth of all three species is due to lack of oxygen with a consequent reducing action in the soil or water; that this reducing action may be produced by stagnation in the bottoms of ponds or by improper or too rapid filtration in wells or filter galleries. Mt. Prospect Laboratory, Brooklyn, New York City. PLATE X A NEW SPECIES OF CRENOTHRIX 39 EXPLANATION OF PLATE Platz X Fig. 1. Chrenothrix mangantfera. Magnification 800 diameters. The manganese has been removed from the gelatinous sheath in the lower half of the field by means of acid and the bacteria stained with Loeffler solution in order that the filaments and the inclosed bacteria may be clearly shown. Near the lower edge of the field may be seen a case of multiplication by trans- verse division. The thread-like attachment connecting each bacterium with the next has probably never before been photographed and can not be shown except by pho- tography. This photograph and Nos. 4 and 6 were taken from the author’s preparations by Dr. R. B. Fitz-Randolph of the Hoagland Laboratory, Brook- lyn, N. Y. Fig. 2. Crenothriz kihniana. Magnificstion 250 diameters. In this photograph the filaments and bacteria are entirely covered by the precipitated iron in the gelatinous sheath. Nos. 2 and 5 are from photographs by G. C. Whipple. Fig. 3. Crenolhriz kihniana. Magnification 1,200 diameters. A young growing filament is shown in the upper part of the field, while in the lower part is a filament filled with microspores. Photograph by George W. Rafter. Fig. 4. Crenothrix ochracea. Magnification 800 diameters. The filaments may be seen through the translucent precipitated alumina in the sheath. Fig. 5. Crenothrix kihniana. Magnification 800 diameters. The iron has been nearly all dissolved by acid and the filmaments with their articulatious may be seen. Fig. 6. Crenothrix mangantfera. Magnification 800 diameters. The manganese has been dissolved by acid and the articulated filaments are distinctly shown. Note the variation in size and general appearance in the three species rep- resented by Nos. 4, 5, and 6. NOTES ON COLORADO ENTOMOSTRACA By ARTHUR E. BEARDSLEY The entomostracan fauna of Colorado, although of great economic importance as the chief source of food supply for the fishes of the state, has hitherto received but little attention. The early explorers noted a single species which they found in great numbers in tem- porary pools of rain water on the plains to the east of the moun- tains, and the naturalists connected with the geological surveys during the ’7os found several species in the elevated mountain lakes and one species on the western slope. For more than half a century no addition has been made to the entomostracan fauna of the great plains of eastern Colorado. From the geographical position occupied by the state, in the midst of the great arid region, it might be inferred that entomos- tracan life, dependent as it is upon the presence of a body of water for its very existence, would be but poorly represented. The results obtained in the preliminary work which it is the purpose of this paper to record would indicate that it is abundant both in species and individuals. That many species of Entomostraca are capable of producing eggs which may remain dormant through long periods of drouth, to be afterwards awakened into activity upon being supplied with water in sufficient quantity for their development, is a fact which has been frequently demonstrated. This may be true of most, if not all, of the forms commonly found in fresh waters. Their rapid development and great fecundity under favorable conditions of en- vironment make it possible for their life cycle to be completed and a new supply of eggs to be deposited in a temporary pool, which may be dried up in the course of a few days or a few weeks at most. When it is remembered that the arid region is subject to occasional rainstorms of great violence, locally known as “cloud- bursts,” and that the snows of winter, scanty though they be, may furnish sufficient water to fill the small depressions in the plains, 42 ARTHUR E. BEARDSLEY and also that, owing to the aridity of the climate and the conse- quent lack of stream erosion, such depressions as are capable of holding water are to be found in great numbers on the plateaus of the arid region, it will be readily perceived that conditions favor- able to the development of the Entomostraca are not wanting. HISTORICAL The earliest reference to Entomostraca in what is now the state of Colorado appears in the report of Long’s expedition, in which James (23)! notes the occurrence of a species of Apus, which he calls A. obtusus, in “rain-water puddles on the Platte river near the Rocky mountains.” Unfortunately, his description is so meager that the species is not recognizable. Packard (74a) thought it “probably the same” as Le Conte’s 4. longicaudatus. Twenty-three years later, Le Conte (46) described Apus longi- caudatus, which, he says, was “found in immense numbers in a small shallow lake on the high plateau between Lodge Pole creek and Crow creek, northeast of Longs Peak.” Nearly thirty years after Le Conte, Carpenter (74) noted the occurrence of Daphnia pulex in a pool above timber-line on Mt. Elbert, and Packard described (74) a lernaean (Achtheres Carpen- teri) and (74a) a new branchiopod (Branchinecta coloradensis) from the mountains of Colorado. Three years later he described (Packard 77) another branchiopod (Lepidurus bilobatus) from Po cafion, and Chambers (77) recorded the occurrence of two spe- cies of Cladocera (Daphnia brevicauda n. sp. and Chydorus sphaeri- cus), and three new species of Ostracoda (Cypris grandis, C. altis- simus, and C. mons), making eleven species of Entomostraca reported from Colorado up to the close of the year 1877, as follows: Branchiopoda Apus obtusus James. Lepidurus bilobatus Packard. Apus longicaudatus Le Conte. Branchinecta coloradensis Packard. Cladocera Chydorus sphaericus O. F. Miiller. Daphnia brevicauda Chambers. Daphnia pulex De Geer. Ostracoda Cypris grandis Chambers. Cypris mons Chambers, Cypris altissimus Chambers. Copepoda Achtheres Carpenteri Packard. 1See list of works cited at the end of this paper. NOTES ON COLORADO ENTOMOSTRACA 43 Since 1877 no additions to the list appear to have been reported. Within the past year the writer has made a large number of col- lections of Entomostraca in the vicinity of Greeley, Col., and these collections, together with a few others made in former years in various parts of eastern Colorado, form the basis of the present paper. BRANCHIOPODA LIM NADIADAE Eulimnadia texana Packard. I found this species in a muddy pool in Crooked creek, Otero county, in June, 1882, where it occurred in great numbers together with the following species. It is also found in Texas and Kansas (Packard, 83). Estheria mexicana Claus. I collected about twenty individuals of this species from the same pool with the preceding. I found it again in August, 1897, in Little Crow creek, in Weld county. It has been reported from Mexico, New Mexico, Kansas, Lake Winnipeg, Ohio, and Ken- tucky (Packard 83). APODIDAE Apus Newberryi Packard. Occurred abundantly in a pool in Little Crow creek in August, 1897, together with the preceding. This species has been found hitherto only in Utah; its occurrence on the eastern side of the mountains is a matter of interest, as it is the first instance in which the same species of crustacean has been found in both regions. BRANCHIPODIDAE Branchinecta Lindahli Packard. I found a single specimen of this species, a female with eggs, in a temporary pool near Greeley, July 2, 1901, together with Moina affinis. Hitherto its only recorded habitat was Kansas (Packard 83). Streptocephalus texanus Packard. About thirty individuals of this species, including females with eggs and adult males, were collected by the writer from a rock pool 44 ARTHUR E. BEARDSLEY filled with water from melting snow, in April, 1882, on the north side of the Mesa de Mayo. It has been reported from Texas and Kansas (Packard 83). CLADOCERA DAPH NIADAE Daphnia pulex De Geer. Occurs in Seely lake; also occasionally in pools about Greeley. Ceriodaphnia reticulata var. dentata Birge. Abundant in Seely lake and in ponds around Greeley. Scapholeberis mucronata (O. F. M.) Of frequent occurrence in ponds near Greeley. Simocephalus vetulus (O. F. M.). Abundant around Greeley in all ponds. Moina affinis Birge. Often extremely abundant in pools formed by summer rains. LYNCEIDAE Alona glacialis Birge. Occurs sparingly in ponds and in Seely lake. Alonopsis latissima Kurz. In ponds near Greeley. Not common. Chydorus sphaericus (O. F. M.). Occurs in the majority of my collections from the ponds about Greeley ; also from Seely lake. OSTRACODA CYPRIDIDAE Candona acuminata (Fischer). I found this occurring in great numbers in March and April, in a small grassy pool near Greeley, which soon after became dry. Cyclocypris laevis (O. F. M.). I have found this species in shallow ponds near Greeley in Febru- ary and March. NOTES ON COLORADO ENTOMOSTRACA 45 Cypridopsis Newtoni Brady and Robertson. Common in ponds near Greeley in June and July. Cypridopsis vidua (O. F. M.). Abundant during the summer in stagnant pools. Cypris fuscata Jurine. I have found a few individuals of this species in Carter’s slough near Greeley. Erpetocypris olivacea Brady and Norman. Occurs in abundance in Carter’s slough. COPEPODA CENTROPAGIDAE Diaptomus sicilis Forbes. Occurs in Seely lake, where it is the most abundant of the Entomostraca. Diaptomus clavipes Schacht. I found this species occurring in a pool of only a few feet area in a narrow ravine fed by springs. My specimens when alive were transparent and colorless excepting the distal portion of the an- tennae, which were blood red. The peculiar hook on the fifth foot of the male is represented in its correct position in Schacht’s figure, but in his description it is erroneously placed upon the next segment. CYCLOPIDAE Cyclops insectus Forbes. C. albidus Jurine. C. ater Herrick. C. serrulatus Fischer. Excepting C. ater, which is apparently rare, these are abundant in the regions about Greeley. HARPACTIDAE Canthocamptus minutus (O. F. M.) Length of female, 0.65-0.75 mm. Length of male, 0.60-0.65 mm. My specimens were obtained from a small pond near the track of the C. & S. railroad in the city of Greeley. 46 ARTHUR E. BEARDSLEY LIST OF ENTOMOSTRACA KNOWN TO OCCUR IN THE STATE OF COLORADO (An asterisk [*] placed before the name of a sp2cies indicates that it is new to the state. ) BRANCHIOPODA Limnadiadae * Eulimnadia texana Packard. * Estheria mexicana Claus. A podidae Apus longicaudatus Le Conte ( ?= * Apus Newberryi Packard. obtusus James). Lepidurus bilobatus Packard. Branchipodidae Branchinecta coloradensis Packard. * Streptocephalus texanus Packard. * Branchinecta Lindahli Packard. CLADOCERA Daphniadae Daphnia pulex De Geer. *Scapholeberis mucronata (O. F. Daphnia brevicauda Chambers. Miller). * Ceriodaphnia reticulata var. dentata *Simocephalus vetulus (O. F. Muller). Birge. * Moina affinis Birge. Lynceidae * Alona glacialis Birge. Chydorus sphaericus (O. F. Miil- * Alonopsis latissima Sars. ler). OSTRACODA Cyprididae * Candona acuminata (Fischer). Cypris altissimus Chambers. *Cyclocypris laevis (O. F. Muller). * Cypris fuscata Jurine. Cypria mons (Chambers). Cyprinotus grandis (Chambers), *Cypridopsis Newtoni Brady and *Erpetocypris olivacea Brady and Robertson. Norman. * Cypridopsis vidua (O. F. Miiller). COPEPODA Centropagidae * Diaptomus sicilis Forbes. * Diaptomus clavipes Schacht. Cyclopidae * Cyclops insectus Forbes. * Cyclops ater Herrick. * Cyclops albidus Jurine. * Cyclops serrulatus Fischer. Harpactidae *Canthocamptus minutus (O. F. Miller). Lernaeopodidae Achtheres Carpenteri Packard. NOTES ON COLORADO ENTOMOSTRACA SUMMARY Branchiopoda 8 species, of which 5 are new to the state. Cladocera 9 species, of which 6 are new to the state. Ostracoda 9 species, of which 6 are new to the state. Copepoda 8 species, of which 7 are new to the state. Total, 34 species, of which 24 are new to the state. Biological Laboratory Colorado State Normal School. 47 48 ARTHUR E, BEARDSLEY WORKS CITED CARPENTER, LikuT. W. L. 74. Report on the Alpine insects fauna of Colorado, 74h Ann. Rept. U.S. Geol. Surv. (Hayden), 539-42. 1874. CHAMBERS, V. T. 77. New Entomostraca from Colorado, Bull. U.S. Geol. Surv. (Hayden), III, 151-55, figs. 1-4. 1877. JAMES, EDWIN. 23. Note 7 [on the appearance of Crustacea] to Long’s expedition from Pittsburg to the Rocky mountains, II, 336. 1823. LE CONTE, JOHN L. 46. Description of a new species of Apus, Annals New York Lyceum Nat. Hiist., 1V, 155. 1846. PACKARD, A. S., JR. 74. Description of a Lernaean Crustacean obtained by Lieut. W. L. Carpen- ter in 1873 in Colorado, 7th Ann. Rept, U. S. Geol. Surv. (Hayden), 612. 1874. 74a. Synopsis of the fresh-water Phyllopod Crustacea of North America, 7th Ann. Rept. U. S. Geol. Surv. (Hayden), 613-22, pl. I-IV. 1874. 77. Description of New Phyllopod Crustacea from the West. Bull. U. S. Geol. Surv. (Hayden), III, 171-79, figs. 11-14. 1877. 83. Monograph of the Phyllopod Crustacea of North America, with re- marks on the order Phyllocarida, z2th Ann. Rept. U. S. Geol. Surv. (Hayden), IT, 295-593, pl. I-XXXIX. 1883. NOTES ON COLORADO PROTOZOA WITH DESCRIPTIONS OF NEW SPECIES By ARTHUR E. BEARDSLEY WITH ONE PLATE These notes have been made in the course of a systematic study of the Protozoa of the state, which was begun in the spring of 1897 and has been carried on at such times as the pressure of other duties has permitted. The work up to the present time has been confined to Greeley and its immediate vicinity, but it is hoped to extend it in the near future to other parts of the state. Class SARCODINA Order RHIZOPODA Family AMOEBAEA LOBOSA Genus AMOEBA Amoeba spatula Penard. Pl. XI, figs. 1a-te. A very small, floating Amoeba with long, radiating pseudopods is frequently to be found in cool, shaded waters about the city of Greeley. I have found it most abundant during the winter months in a large tank at the edge of the city park, used as a horse-trough and supplied with water from the city mains. At first I supposed it to be Ehrenberg’s A. radiosa, and cast aside any lingering doubts that may have remained when I found it well represented by several figures in Leidy’s Fresh-water Rhizopods under that name. From the same place, and often on the same slide with the fore- going, I found a small, reptant Amoeba (fig. 1a) of slow movement, 50 ARTHUR E. BEARDSLEY in outline broadly spatulate or fan-shaped, its width equal to or ex- ceeding its length, the anterior half strongly depressed, very broadly rounded, thin, and hyaline, forming a sort of broad, flat pseudopod; posterior portion thickened, granular, and filled with food vacuoles. Contractile vesicle usually one, sometimes two or three, filling and emptying very slowly. Nucleus small, round, rarely visible without reagents. This form is apparently identical with A. spatula Penard. Subsequent observations proved conclusively that the two forms are merely different states of the same organism. Individuals were repeatedly seen to change from one form to the other. When about to change from the spatulate to the radiate form, the Amoeba projects from its posterior, thickened portion one or two slender pseudopods, and at the same time the broad, hyaline anterior por- tion is gradually withdrawn (fig. 1b) ; then several pseudopods are thrown out from what was the anterior end, the animal frees itself from the slide, and floats away (Ic). In changing from the float- ing to the creeping form, this process is reversed: the long, slender pseudopods are withdrawn, the animal flattens itself upon the slide, and assumes the spatulate form. This species attains a diameter of 10-12 », when in the radiate, floating form, and 20-25 , in the spatulate form. In the radiate form it may easily be mistaken for Amoeba (Dactylosphaera) radi- osa, from which it is distinguishable by its habit of transformation. Family Gromuna Bitschli Pamphagus mutabilis Bailey. A few individuals of this apparently rare form have been seen in water from the horse-trough in the city park. Sub-class HELIOZOA Genus NUCLEARIA Nuclearia delicatula Cienk. (Syn.—Heterophrys varians F. E. Schulze.) This interesting heliozoan has been found abundantly in the horse-trough in City Park, and its mode of taking food has been frequently observed. Its food consists chiefly of diatoms and uni- NOTES ON COLORADO PROTOZOA ps cellular green algae. A pseudopod, on coming in contact with a food-particle, adheres to it and slowly contracts; as the adhering particle approaches the body, a small rounded vesicle rises up around the base of the pseudopod, into which the particle is drawn by the retraction of the pseudopod; after the particle is completely enclosed in the vesicle, this gradually subsides, carrying the particle with it into the body. Change of form in Nuclearia, as in most Heliozoa, is usually very slow, but occasionally a complete trans- formation in appearance is effected in a few minutes. Figures of what is probably this species, from the Uinta moun- tains, Wyoming, are given by Leidy in his Fresh-water Rhizo- pods,’ and are doubtfully referred by him to the genus Heterophrys. I have been unable to find any other record of the occurrence of Nuclearia in North America. Class MASTIGOPHORA Family CERCOMONADINA (Kent) Cercomonas longicauda Duj. In water from the horse-trough in City Park, I have frequently found a monad belonging to this genus and which accords with the figures and description of this species. I have been unable to find any prior record of the occurrence of this genus in America. Family Menorpi1na Bitschli Atractonema teres Stein. Body colorless, rigid, fusiform, pointed behind, truncate or slightly emarginate anteriorly. Mouth at the anterior end opening into a distinct oesophagus which is enlarged posteriorly. Flagellum single. My specimens agreed in all respects with Stein’s figure. Hab.—Pond water, with Euglena. Apparently not hitherto re- corded from America. Family AMPHIMONADINA Kent Genus Amphimonas Duj. Amphimonas clavatan. sp. Pl. XI, figs. 2a, 2b. Form elongate-clavate, broadly rounded anteriorly, tapering grad- ually to the posterior extremity by which it frequently fixes itself 1 Leidy, Fresh-water Rhizopods, pl. XLV, figs. 2, 3, 5, 6. 52 ARTHUR E. BEARDSLEY to other objects. Body somewhat contractile. Flagella two, similar in size and character, nearly as long as the body, arising close to- gether from the anterior extremity. Endoplasm colorless, slightly granular. Contractile vesicle single in the anterior half. Nucleus spherical, sub-central. Length 8-10 up. Hab.—Stale pond water. This species closely resembles Deltomonas cyclopum Kent, but differs in the point of origin of the flagella. It apparently is near to A, exilis Perty, but differs in the flagella, which are less than half as long as in exilis. Family CHLAMYDOMONADINA Bitschli Spondylomorum quaternarium Ehrbg. Colonies consisting of sixteen green monads, agreeing in all re- spects with Stein’s figure and description, were abundant in June, 1898, and have since been seen occasionally. | Occurs in Europe and India; not hitherto reported from America. Sub-class DINOFLAGELLATA Biitschli Ceratium hirundinella (O. F. M.). I have found this occasionally, but only in small numbers. Peridinium tabulatum Ehrbg. Common in clear, open water in ponds and lakes. I have found it in Seely lake, near Greeley, sometimes occurring in long streaks through the water, so abundant as to give to the water a well- marked, reddish-brown tint, perceptible at a distance of twelve to twenty meters. An average of ten counts from such a streak gave 18,300 Peridinia per cubic centimeter. NOTES ON COLORADO PROTOZOA 53 Class INFUSORIA Order GYMNOSTOMATA Biitschli Family ENCHELINA Ehrbg., Stein Genus HoLopHrya Ehrbg. Holophrya heterostoma n. sp. Pl. XI, fig. 3. Form ellipsoid, about twice as long as wide; faintly striate longi- tudinally with about twenty striae; cilia fine, short, nearly equal, those about the anterior pole slightly larger. Mouth a narrow oval slit, rounded in front, pointed behind, lying at one side of the an- terior pole. Color translucent. Usually filled with large food- balls. Nucleus oval, single, sub-central. Contractile vesicle single, posterior. Movement rather slow, regular. Length, 100 p. Hab.—Ponds and ditches. This species is a member of Section II of the genus as arranged by Butschli. It may be readily distinguished from H. Lieberkuhnu Biitschli, the only other member of this section, by the superficial striae which are much more numerous in the latter. Butschli gives no description of his species, but merely a figure of the anterior pole, which he copies from Lieberktthn; its remaining character- istics are therefore uncertain. Genus URoTRICHA. Urotricha farcta C. and L. This species has apparently not hitherto been reported from America, but my specimens were so completely in agreement with the description and figures by European authors as to leave no doubt of their identity. One evening, while observing through the microscope, I discov- ered a double Urotricha, a most diminutive pair of “Siamese twins.” The two individuals were grown together, side by side, their long, posterior cilia extending backward parallel one with the other. Aside from the union, each appeared to be quite normally developed in every way. They were kept under observation nearly an hour, and were finally lost by their suddenly springing out of the field. 4 54 ARTHUR E. BEARDSLEY Since, in this family, reproduction commonly takes place in the encysted state, it seems probable that the “twinning” in this case was due to arrest of the process of division between two of the segments while in the cyst. Prorodon teres Ehrbg. I have occasionally seen infusoria which agree in every respect with Biitschli’s figures of this species. It has apparently not been heretofore reported from America. Lagynus laevis (Engelm.) ? I have found infusoria in Brown’s slough, near Greeley, which apparently belong here, although less than half the size indicated by Engelmann’s figure. They occurred in considerable numbers and some were found in conjugation. Didinium nasutum (O. F. M.) Didimium Balbiant Biutschli. Both of these occur in abundance in Brown’s slough. Neither appears to have been hitherto reported from America. Nassula aurea Ehrbg. This highly colored infusorian, resplendent with royal purple and gold, is often found in shallow, quiet water in the Cache la Poudre river. Apparently not hitherto reported from America. Order TRICHOSTOMATA Biitschli Family CHILiFrera Biitschli Genus Frontonia C. & L. Frontonia leucas Ehrbg. Common in ponds, among diatoms and algae. Apparently not hitherto reported from America. Frontonia elliptica n. sp. Pl. XI, figs. 4a—4d. Form ellipsoid, slightly flattened; ends equally rounded. Body flexible, not contractile, covered with fine longitudinal striations. Cilia fine and even. Trichocysts numerous, evenly distributed. Mouth and post-oral groove extend along the second and third fifths of the body; mouth about half covered by the left undulating membrane. Contractile vesicles two, with distinct afferent radi- NOTES ON COLORADO PROTOZOA 55 ating canals. Anus postero-lateral. Macro-nucleus large, spherical, lying in the anterior half, with one or more imbedded micro-nuclei. - Feeds upon diatoms. Length 115-150 yp. Hab.—Bottom of ponds, with algae and diatoms. This species differs from /eucas, with which I have found it some- times associated, in its more symmetrical form and smaller size, in the constant presence of two contractile vesicles, in the shape of the nucleus, and in the form of the mouth and the left undulating membrane. In most of these characters it resembles F. fusca Quen- nerstedt, a marine species, from which it differs in the form of the nucleus and the position of the mouth. Family MicroTHoracina Wrzesn. Cinetochilum margaritaceum (Ehrbg.) (Syn.—Cyclidium margaritaceum Ehrbg.) I have found this minute infusorian very common among algae in ponds and streams. Apparently not hitherto reported from America. Sub-order SPIROTRICHA Biitschli Section HETEROTRICHA Stein Family Gyrocor1na Stein Coenomorpha medusula Perty. (Syn.—Gyrocoris oxyuris Stein.) This rare infusorian occurred in great numbers in a jar with Lemna in my laboratory, in November, 1897. Section OLIGoTRICHA Biitschli Family HatTertna C. & L. Genus StrompBipium C. & L. Strombidium velox n. sp. Pl. XI, figs. 5a—5c. Form turbinate, varying to obovate and broadly elliptical. Peris- tome produced backward along the ventral side as an oblique furrow, nearly to the middle line. Adoral cilia stout, curved outward and 56 ARTHUR E. BEARDSLEY backward, about half as long as the body, diminishing in size toward the posterior end of the oral groove. Body colorless; surface smooth, without supplementary cilia. Nucleus sub-central, irregu- larly globular. Contractile vesicle anterior, spherical, rather large. Food consists of diatoms. Length, 45-50 wu. Greatest width, 37-40 mp. Hab.—Ponds and ditches, with Vaucheria. The movements are extremely rapid and erratic, the infusorian frequently gyrating for a time around a fixed point, then suddenly darting away and out of sight, even when one is using a low power. It often remains for a considerable time fixed by a slender, color- less filament to the glass or to debris on the slide, rapidly revolving on a longitudinal axis and swaying to and fro with a pendulum- like movement, in a manner quite similar to that of Urocenirum turbo. The filament by means of which the animal is enabled to fix itself is upparently formed of some gelatinous substance, and is probably a secretion of the posterior portion of the animal itself. That it possesses a considerable amount of resilience and tenacity is shown by the way in which the animal is drawn backward by its contrac- tion whenever a slowing up of the ciliary motion occurs, as well as by the comparatively enormous loads of debris which are occa- sionally drawn by it across the field. A number of forms have been found which have not yet been sufficiently studied to make their identification complete; of these no further mention is made at this time. The following list con- tains only those forms which I have found occurring within the state and have fully identified. Of the 99 species recorded in this list, 4. are apparently new; 13 are Old World species which do not appear to have been hitherto reported as occurring in North Amer- ica; the remaining 82 species embrace a few rare forms not often seen, but the greater number are well-known forms, many of them cosmopolitan in distribution. NOTES ON COLORADO PROTOZOA 57 A PRELIMINARY LIST OF THE PROTOZOA FOUND IN THE STATE OF COLORADO (An asterisk [*] before the name of a species indicates that it is new to America; the dagger [}] indicates a species new to science. } Class SARCODINA Order RHIZOPODA Amoeba proteus (Rosell). Amoeba limax Duj. Amoeba spatula Penard. Amoeba radiosa Ehrbg. Arcella vulgaris Ehrbg. Arcella discoides Ehrbg. Difflugia acuminata Ehrbg. Difflugia constricta (Ehrbg.). Difflugia corona Wallich. Difflugia globulosa Duj. Difflugia lobostoma Leidy. Difflugia pyriformis Perty. Centropyxis aculeata (Ehrbg.). Nebela flabellum Leidy. Euglypha alveolata Duj. Cyphoderia ampulla (Eurbg.). Pamphagus mutabilis Bailey. Order HELIOZOA Vampypella lateritia (Fresenius). Nuclearia delicatula Cienkowsky. (Heterophrys ? Leidy.) Actinophrys sol Ehrbg. Actinosphaerium Hichhornii Ehrbg. Raphidiophrys pallida F. E. Schulze. Class MASTIGOPHORA * Cercomonas longicauda Duj. Oikomonas mutabilis Kent. Oikomonas termo (Ehrbg.). Monas guttula Ehrbg. Anthophysa vegetans (O. F. M.). Euglena viridis Ehrbg. Trachelomonas volvocina Ehrbg. Trachelomonas hispida Perty. Phacus pleuronectes (Ehrbg.). * Atractonema teres Stein. Peranema trichophorum (Ekhrbg.) Stein. (Astasia trichophorus. ) Anisonema ovata Duj. { Amphimonas clavata n. sp. Synura uvella Ehrbg. Mallomonas Plosslii Perty. *Spondylomorum quaternarium Ehrbg. Gonium pectorale (O. F. M.). * Gonium sociale (Duj.). Pandorina morum Ehrbg. Eudorina elegans Ehrbg. Chilomonas paramaecium Ehrbg, Ceratium hirundinella (O. F. M.). Peridinium tabu’atum (Ehrbg.). Class INFUSORIA (CILIATA) Order GYMNOSTOMATA Bitschli Holophrya discolor Ehrbg. + Holophrya heterostoma n. sp. *Urotricha farcta Claparede Lachman, and * Prorodon teres Ehrbg. Lacrymaria olor Ehrbg. (Trachelocerca olor. ) * Lagynus laevis (Eng-lmann), 58 ARTHUR E. BEARDSLEY Trachelophyllum apiculatum Lionotus fasciola (Ehrbg.). (Perty). Dileptus anser (O. F. M.). Coleps hirtus Ehrbg. | Loxodes rostrum (O. F. M.). *Didinium nasutum (O. F. M.). * Nassula aurea Ehrbg. * Didinium Balbiani Butschli. Chilodon cucuilus (O. F. M.). Lionotus anser (Ehrbg.). Order TRICHOSTOMATA Sub-order ASPIROTRICHA Biitschli Glaucoma scintillans Ehrbg. Paramaecium caudatum Ehrbg. * Frontonia leucas Ehrbg. Urocentrum turbo (O. F. M.). { Frontonia elliptica n. sp. Lembadion bullinum Perty. Colpidium colpoda Ehrbg. (Hymenostoma hymenophora Colpoda cucullus (O. F. M.). Stokes. ) *Cinetochilum margaritaceum Cyclidium glaucoma Ehrbg. (Ehrbg.). Sub-order SPIROTRICHA Biitschli @ HETEROTRICHA Euplotes patella (O. F. M.). Euplotes charon (O. F. M.). Blepharisma lateritia Ehrbg. ASpitlices Lanes (gee Metopus sigmoides Cl. and L. Spirostomum ambiguum Ehrbg. % PERITRICHA Bursaria truncatella O. F. Mueller. Vorticella aperta Frommontel. oF 2 Vorticella convallaria Ehrbg. ee ee ee Vorticella hamata Ehrbg. Coenomorpha medusula Betty. Vorticella microstoma Ehrbg. Se ee Vorticeila nebulifera Ehrbg. 2 OLIGOTRICHA Vorticella putrina O. F. M. Vorticella similis Stokes. Carchesium polypinum Ehrbg. Epistylis digitalis Ehrbg. 7 Strombidium velox n. sp. Halteria grandinella (O. F. M.). % HYPOTRICHA Cothurnia crystallina (Ehrbg.). * Oxytricha pelionella (O. F. M.). Thuricola valvata (Wright). Urosoma caudata (Stokes). Vaginicola decumbens Ehrbg. Stylonychia mytilus (O. F. M.). Sub-class SUCTORIA Sphaerophrya magna Maupas. Grand “Total 25233. Pee ee ee ot eee ee Biological Laboratory, Colorado State Normal School. RLATE XI NOTES ON COLORADO PROTOZOA 59 EXPLANATION OF PLATE Plate XI All figures (excepting 14, lc, and 54) were drawn with aid of a camera lucida; the linear magnification of each figure is indicated by the number fol- lowing the sign X. In all figures the letters cv indicate the contractile vesicle; /, ingested food; and , the nucleus. Fig. la-e. Amoeba spatula Penard. a. Reptantform, x700. dandc. Suc- cessive stages of the same individual, changing from the reptant to the floating form, < about 600. d. Floating form with pseudopods partially extended, 600. e. Same, pseudopods fully extended, 600. Fig. 2, a, 6. Amphimonas clavata n.sp. a. Free-swimming individual, 1300. 6. Individual attached by posterior end, 1300. Fig. 3. Holophyra heterostoma n. sp. Ventral view, 260. Fig. 4a-d. Frontonia elliptican. sp. a. Ventral view; ¢, trichocysts; 200. 6. Oral aperture, with wm, the undulating membrane; 480. c. Outline, showing the two contractile vesicles with their afferent canals. d. Outline, showing 2, the macro-nucleus with n’, the imbedded micro-nucleus (stained with acetic acid carmine). Fig. 5a-c. Strombidium velox n.sp. a. Ventral view of a quiescent indi- vidual with rounded posterior end, 370. 6. Dorsal view of turbinate indi- vidual, showing mode of attachment. c. Isolated nuclei (stained with acetic acid carmine); 600. ’ | yh) , 5 NS UNE Wi Us AY Path a a j i et ho} DAN PA ne A DAN! eae } f Wray THE PLANKTON OF LAKE MAXINKUCKEE, INDIANA By CHANCEY JUDAY It is desirable to present here a brief summary of the plankton observations made on Lake Maxinkuckee during August, 1899. These observations were made as a part of the field investigations of the U. S. Fish Commission. The following results were obtained: 1. Regular observations were made during August at three dif- ferent stations and these showed a decrease in the quantity of plankton of from 36 per cent to almost 50 per cent. This decrease was due to a large decrease of the phytoplankton. There was a slight increase of crustacea. Cyclops was the predominant crustacean. 2. The plankton was confined almost wholly to the upper 12 meters. Only Corethra larvae were found regularly below this depth. The o-1 m. layer contained about 48 per cent of the entire quantity. This is preeminently the region for phytoplankton, young Cyclops, and nauplii. The o-3 m. layer contained about 68 per cent of the entire quantity of plankton. 3. Adult Diaptomus minutus and Daphnia retrocurva were rarely found above 3 m. during the day. Daphnia pulex was found in and below the thermocline. 4. Only one set of observations was made at night. Although the entire quantity of plankton in the o-1 m. layer was smaller than that obtained during the previous afternoon, there was a distinct in- crease in the crustacea. Epischura lacustris, Leptodora hyalina, and adult Daphnia retrocurva were found in the o-1 m. layer at night but not during the day. Also, there was an increase in the number of adult Cyclops over day conditions. 5. Station II was located in a small basin having a maximum depth of 12.8 m. and entirely separated from the main basin of the lake by a considerable area where the water is 2 m. or less in depth. The bottom temperature was slightly lower here than in the deepest 62 CHANCEY JUDAY part of the lake (26 m.). During each of the four weeks ee ob- servations continued, one set of hauls was made in this small basin for purposes of comparison with hauls from Station I in the deep- est part of the lake. The quantity of plankton obtained was smaller than the weekly average from corresponding depths at Station I. Hauls made at five stations in the main basin of the lake, besides the two regular stations, showed that the plankton was very evenly distributed. The following is a list of the Cladocera observed : Daphnia pulex De Geer, var. puli- Pleuroxus procurvatus Birge. caria Forbes. Diaphanosoma brachyurum Liéyin. Daphnia retrocurva Forbes. Ilyocryptus longiremis Sars. Ceriodaphnia lacustris Birge. Alona guttata Sars. Sida crystallina O. F. Miiller. Leptodora hyalina Lilljeborg. Acroperus harpae Baird. Prof. C. Dwight Marsh determined the following list of Copepods observed : Cyclops Leuckarti Sars. Diaptomus minutus Lilljeborg. Cyclops prasinus Fischer. Epischura lacustris Forbes. THE MORPHOGENESIS OF, THE STIGMATA AND STOMATA OCCURRING IN PERITONEAL AND VASCULAR ENDOTHELIUM By ARTHUR E. HERTZLER, A.M., M.D. WITH TWO PLATES PREFATORY NOTE Although a great many papers have been written which, either directly or indirectly, involve a discussion of the stigmata and sto- mata, their significance has never been definitely determined. The majority of the discussions have to do, not with these structures themselves, but with their influence and importance in certain physi- ologic and pathologic processes. Their existence as real morpho- logic elements has been assumed and then their function or office argued on a priori grounds. This method of study is always un- certain and doubly so when morphologic problems are under discussion. The study of which this paper is an excerpt was begun nearly ten years ago, when the writer was still in his undergraduate course. It at first involved surgical problems only, but it soon became evi- dent that anatomic knowledge was inadequate for successful study of surgical and pathologic problems. A review of the literature gave but little aid and less encouragement, for it was apparent that the history of our knowledge of the structure of the peritoneal endo- thelium? is coincident with that of the action of the nitrate of silver upon this structure. A reconsideration of the problem with the aid of modern technical methods seemed imperative. In order to meet this requirement the writer has tried to bring to bear on this prob- lem the fruits of modern perfected histologic technic. 1 Waldeyer advises (Arch. f. mikroskop. Anat., Bd. 57, Heft I, S. 4, 1900) that the term ‘‘en- dothelium ”’ be retained for the cells lining the blood and lymph vessels and the chambers of the eye only. In this paper, as a matter of convenience, the advice of this eminent authority att Be epeatded and the term endothelium will be used according to the original sugges- ion of His. 64 ARTHUR E. HERTZLER The present communication has to do with the so-called stigmata and stomata only (as occurring between endothelial cells) and dis- cussion will be confined to these, with casual reference to other facts determined by these studies when they serve to illustrate the points under discussion. Most of the work was carried on far removed from most of the advantages desirable for this kind of work. This misfortune has been especially felt in reviewing the literature, and the writer re- grets to state that he has been unable to confirm, at the last moment, his citations. Since some of the papers were read years ago, only notes were available at this time, and many errors will probably have been committed. It is pleasing to state, however, that a part of the work was done in the laboratory of Dr. H. Virchow, Professor in Berlin. It is with pleasure that the writer takes this opportunity of expressing his appreciation of the many kindnesses received while doing this part of the work. ‘The obligation presents a wider range than can be mentioned and represents every favor that a teacher can bestow upon a pupil. The writer is also indebted to Dr. Kopsch for many suggestions in technic.? CITATION OF LITERATURE As is well known, it was Coccius and Flinzer (1) who first used the nitrate of silver to demonstrate the cell outlines of Descimet’s membrane. This method was perfected by His (2). The method was still further developed by v. Recklinghausen (3) and used by him in his study of the lymphatic system. It was while using this method, in the study of the peritoneum, that Oedmasson (4) first noticed the occurrence of small dots and rings in the intercellular lines. The discovery was accidental, for, as the title of his paper indicates, he was concerned only with a review of v. Reckling- hausen’s work. Oedmasson himself hardly regarded the discovery seriously, but was rather inclined to believe that the rings were real openings because of their regularity. The stigmata he regarded as deposits of silver albuminate because of their irregular size and form. Not until after the same structures were observed by His (5) 2 Professors Sayer and Bailey. respectively heads of the departments of pharmacy and themistry in the University of Kansas, have ren lered aid in the consideration of chemical problems. To them also the writer expresses appreciation. STIGMATA AND STOMATA OF THE PERITONEUM 65 in the lymph vessels was Oedmasson’s discovery regarded as of im- portance. From then on the belief in their entity gradually became general. A number of other papers were written about this time, the most important of which were those of Ludwig and Schweigger- Seidel (6), Dybkowsky (7), and Schweigger-Seidel and Dogiel (8), none of which added greatly to our knowledge of the subject in advance of that presented in previous publications. The highest interest in the stigmata and stomata was not reached, however, until Cohnheim (g) described them in the blood-vessels and used them as important elements in his theory of infiammation.* It is to this fact, no doubt, that these structures owe much of their prominence, for after their acceptance by Cohnheim their existence was almost universally regarded as proven, though no additional evidence had been adduced to strengthen the position. That the existence of these structures as morphologic elements was never established upon a sound histologic basis may be gathered from the fact that very soon aiter they were first described there were not wanting investigators who regarded them as artificial prod- ucts, due to the reagent used. Among the earlier writers to express their opposition may be mentioned Auerbach (10), Afanassiew (11), Foa (12), Klein (13), and Schweigger-Seidel (1. c.). The last two authors regarded them as real openings, but declared against the function ascribed to them on the ground that no opening could be demonstrated in the basement membrane (membrana limitans) without which the stomata would necessarily be functionally useless. Later on Klein seems to have changed his view somewhat (14 and 15), ascribing more importance to the stomata, even going so far as to class them in two groups, stomata vera and pseudo-stomata. In the former group he reckons only those occurring in the center of groups of radiating cells. To the list of opponents must be reckoned Tourneaux (16). He is very emphatic in his opposition, declaring without qualification that they are artefacts. He based his argu- ment on the physiologic fact that starch and carmine granules are not absorbed when an emulsion of them is introduced into the free peritoneal cavity. These papers were written about the time Cohnheim announced his theory of inflammation. So brilliant were his discoveries, and 8 He accepted the views as expressed by Stricker and Federn as correct (Wien. Acad. Sttzungsber. math.-nat. wits. Cl., Bd. 53. 66 ARTHUR E. HERTZLER so striking his explanation of the phenomena he observed, that the arguments of the opponents of the theory of the existence of special openings between cells were futile. It seemed so clear that the leucocytes, and above all the red corpuscles, in order to escape from the blood-vessels, required the existence of openings that would allow of their escape. The discovery of Oedmasson was eagerly seized upon as offering the needed explanation. Here it seems necessity added one to her progeny: already the mother of invention, she now became the mother of discovery also. Arnold (17, 18, and 19), by his careful researches, aided greatly in strengthening the position of Cohnheim’s theory and more particu- larly the idea of the passage of the blood-cells through preformed openings. It was he who first observed the blood-cells actually to pass through the preformed openings, thus adding demonstration to theory. Arnold (20) presents a cut showing a leucocyte in the act of passing through one of these openings. This has not been ob- served by any other worker. It was this striking demonstration of Arnold’s more than any other factor, apparently, that despaired the opponents of the theory. Notwithstanding the reputation of the author, however, doubts began to arise as to the correctness of the observation, mostly, no doubt, on account of the extreme difficulty of excluding error in determining the exact point in the vessel wall at which the blood- cell made its escape. So uncertain is this method of determining this point that investigators may well despair of direct disproof. Another paper that has had much weight in influencing opinion is that of Muscatello (21), who approached the subject largely from the physiologic side. He ascribes to the stomata the office of ab- sorption, but declares that they exist only on the centrum tendenium of the diaphragm. Ranvier (22) advanced a most convincing argument against the stomata when he showed that their occurrence could be much re- duced in number by first rinsing the surface to be treated with silver solution with distilled water. He also added the additional im- portant argument that their irregular distribution counted much against their functional importance. Whatever opinion one may hold as to the cause of it, the fact remains that the stomata no longer play the part they formerly did in the explanation of physiologic and pathologic processes. To such STIGMATA AND STOMATA OF THE PERITONEUM 67 an extent is this true that the literature of the last decade, of physi- ology and pathology, may be searched without encountering a single reference to them. This fact might be interpreted to mean that these structures have been abandoned and that the subject no longer offers a fruitful field for investigation. That this neglect has not been altogether universal is shown by several recent papers dealing with the histology of the peritoneal endothelium. ‘The most elaborate and complete of these is by Ko- lossow (23), in which the stomata are restored to their former posi- tion. It may be stated briefly in passing that Kolossow makes a modification in that he regards the endothelial cell as being formed of two parts or layers, a granular part containing the nucleus, and a superficial homogeneous part which he calls the cover plate (Deck- platte). It is between these latter that he regards the stomata as existing, and not beween the cells proper. More recently a paper has appeared by Ussow (24) in which a similar position is taken. He believes that ordinarily they do not exist but are brought about by the contraction of the cell proto- plasm. In the short abstract at my disposal no reasons are given for this conclusion. In practical works the stomata stil! play a more important role than among scientific workers. This is well shown by an excellent practical treatise on the peritoneum (27) in which the stomata have ascribed to them their former importance. The other side of the question has recently been strengthened by a very clear paper by Meyer (25). He concludes that the stomata are not real openings. He arrives at this conclusion because of their irregular distribution and because of their irregular size and shape. He believes, however, that they may be artificially pro- duced by mechanical means, though without ascribing to this fact any specific importance. Rawitz* takes a delightful middle ground by saying that if sto- mata do not exist there are at least “soft places.” Nearly all investigators have used the same method. This con- sists, briefly, in treating bits of excised tissue to a solution of silver nitrate and then exposing them to the sun until they become brown. Kolossow (26) alone forms an exception in that he used a solution of osmic acid and then “developed” in a solution of tannin. By 4Grundiss der Histologie. Berlin, 1894. 68 ARTHUR E. HERTZLER this method neither stigmata nor stomata are produced, and his views concerning them are dependent upon the use of silver used according to the usual method. The review of the literature shows that but one method has been used in demonstrating the stigmata and stomata, which suggests that inquiry should be directed quite as much to a criticism of the technic as to the real problem itself. Whether approached by a direct or by an indirect method, the real problem is, Are the stigmata and stomata preformed openings between endothelial cells? If a negative answer is the truth, the question becomes, What is the nature of the so-called stigmata and stomata? An answer to the second question is desirable, not only on account of its interest because of the discussions these structures have occasioned, but because the disposal of them can not be re- garded as final until they have been proven of extraneous origin by direct analysis. FORMS OF STIGMATA AND STOMATA The stigmata have never occasioned any considerable amount of discussion, and they may be considered along with their more prom- inent kin. In order to facilitate the study of the stomata, it is de- sirable to divide the various structures that have from time to time been included in this category into two classes. In the one class may be considered the ordinary form, which may be seen in nearly every cut of the peritoneal endothelium that has been published. In the other group may most conveniently be considered a great variety of different structures that have been regarded as stomata by some observers, or structures incidentally mentioned but which throw some light on the formation of the more common form. The ordinary stigmata and stomata are such familiar structures that a definition is hardly needed, and a satisfactory one is difficult to formulate because it is necessary to regard the opinion of so many different observers. It may, however, in general be stated that the stigmata are small brownish black points, varying in size from a fraction of a micron to ten microns, usually irregular sphe- roids, but often angular, occasionally elongated ovoids, occurring at irregular intervals in the course of the intercellular lines between the endothelial cells, when a serous membrane is treated with silver nitrate and exposed to the sun. By stomata are meant the brownish STIGMATA AND STOMATA OF THE PERITONEUM 69 rings, ovoids, or ellipses, occurring under similar conditions as mentioned above, situated anywhere in the course of the intercellular line, but occurring most typically at the point of juncture of three or more cells. Stigmata and stomata have not been noted, so far as the writer’s knowledge goes, in any other structure than a serous membrane, nor in this structure when treated by any other reagent than nitrate of silver. They may be regarded, then, as the product of two factors ; nitrate of silver and a fresh serous membrane. The formation of these structures was believed to be due to the action of the silver nitrate upon the semi-fluid intercellular cement substance, forming an albuminate of silver. Why a border of the albuminate occurs about the stomata has been explained by the fact that the stomata are formed by a stretching of the membrane, and when the border of the cells retract a part of the cement substance remains adherent which, when acted upon by the silver, forms the stomata. In order to account for the fact that the cells separate only at certain points it was necessary to assume the intercellular substance to be stronger at some places than at others. The relation of the stigmata to the stomata was at first thought by Arnold to be direct, in that he assumed the former to be but early stages of the latter. In his later publications he described them as being but local broadenings of the cement substance—an unskilful distribution of the adhesive, as it were. Most other ob- servers share this view, while a few disregard them entirely. The exact method of formation of the stomata, as has- been stated, has caused considerable controversy. Some regard them as preformed, others as due to the passage of leucocytes, but the ma- jority regard them as due to the stretching of the membrane. The cells were believed not to be quite big enough to cover the entire surface when the membrana limitans is put on the stretch. The writer has made a long series of experiments to determine this point. The result was invariably that, when the membrane was sufficiently stretched to produce a solution of continuity of the cells, they did not separate in the intercellular lines but tore across the cell in the great majority of cases, the direction of separation depending on the line of rupture of the membrana limitans. The cells do not separate so long as the membrana limitans is intact. Even if by chance the cells do separate in the intercellular line it does not occur 5 7O ARTHUR E. HERTZLER at certain points but occurs in a straight line. Kolossow worked out this point and attempted a direct demonstration of the theory. He did this by stretching a serous membrane across the end of a glass tube and then bringing about varying degrees of distention. He found that the stigmata and stomata increased in number as the dis- tention was increased, and in direct proportion. The theory of their formation by the leucocytes was abandoned on the ground that when a vein is tied in which diapedesis has begun it ceases. If due to the activity of the leucocytes the process (of stomata formation) would go on.° If the distention theory of formation were true it would seem . that a great variety of normal stretchings of the serous membranes would produce them and be followed by the usual consequences. But, as is well known, the distentions may be greater under some normal conditions than under some pathologic ones in which diape- desis does occur, without this process taking place. If, in the study of the genesis of the stigmata and stomata, the classic method of treatment, with silver nitrate and exposure to light, be followed, the basis of the whole controversy can be readily ob- served, for all the different pictures described by the various investi- gators may be made out. In reviewing the results the most striking observation is that of Ranvier, already referred to, namely, that the number of stomata may be lessened, and for large areas pre- vented, by a preliminary rinsing with distilled water. So striking is this fact that the probability at once suggests itself that these structures are due to some factor that can be removed with distilled water. That actual openings could be thus removed of course no one would for a moment argue. That large areas may thus be ob- tained free from stomata is a fact various observers have vouched for. Why the cells in such large areas should occur without the ability to perform the same functions as identical cells in other localities is a question Ranvier asked years ago and which still remains unanswered. Observers are likewise agreed that exposure to the sun increases the number of both stigmata and stomata. The same thing is true if the membrane is covered with an excess of lymph.*® 5 Muscatello still adheres to this view, l. c., p. 345. 6Oedmasson (1. c., p. 362) correctly states that the lines are. broader if the strength of the solution is stronger, or allowed to act longer. He also correctly observes that it is due to the action of the silver on the cement substance and not due to its action on the edge of the cell, for the lines may be penciled out. He might have added that this in no wise influences the distance between the edges of the cells, for the increase in width is due to deposits on top of the cells (see figs. 3 and 4, Plate XIII). STIGMATA AND STOMATA OF THE PERITONEUM FE * The problem presenting itself for solution is obviously, from the foregoing, to determine how many stigmata and stomata would remain if all the factors known to cause them were excluded en- tirely, and the factors known to increase them were reduced to a minimum. But with the methods ordinarily employed carried out with the greatest care, one finds himself where he must accept the striking observation of Toldt (28), that “they occur in spite of every precaution, but regarding their significance we are entirely ignorant.” It seems evident from this quotation that advance can be secured only through new methods or by improvements on the old ones. The effect of light is a factor that has never been systematically studied. Handling the tissues in an entirely dark room is difficult and uncertain. An attempt to solve this problem led the writer to develop a new method of using silver. Knowing that Ranvier had injected silver solutions into the adipose tissue in recently killed dogs, in order to demonstrate the fat globules, it required but a step farther to attempt to secure the reaction in vivo. The results ob- tained were gratifying in the extreme, for nothing is simpler than to inject a dilute solution of silver nitrate into the free peritoneal cavity of an animal. The effect of light may thus be determined with a certainty. The anterior abdominal wall of a mouse so treated may be removed in the dark by the aid of the sense of touch and placed upon a slide and the microscope so arranged that the first ray of light that strikes the specimen may be met by the eye of the observer over the microscope. The intercellular lines are seen to be present, thinner and more regular than after the usual method, and stigmata and stomata are not to be found. How much the result may be due to the action of the reagent on living tissue and how much on the exclusion of light must be determined by comparing the results obtained by injecting the solution into the abdomen of a dead animal. For the purpose of the present discussion it makes no difference. The important fact remains: the occurrence of both stigmata and stomata can be entirely prevented. In some animals so treated the usual structures occurred. This is most likely to occur in frogs, especially when the solution is too strong or allowed to act too long. In order to test this method the following animals were injected successively : five new-born mice, six grown but young mice, one pregnant mouse, four new-born rabbits, one old buck 72 ARTHUR E. HERTZLER rabbit, and three kittens six weeks old. In none of them was a single stigma or stoma to be found. In this study it was necessary to remove the parietal peritoneum from the abdominal wall (except in case of the mice and new-born rabbits, in which the abdominal wall was sufficiently transparent to admit of examination in toto) ; the mesentery was also carefully removed, the whole intestinal canal cut open along the mesentric border, after the removal of the mesentery, and examined over its entire extent, and finally the diaphragm and special ligaments were removed as much as possible entire. Of course some nooks and corners escape observation, but with care they may be reduced to a minimum. This series of ani- mals, seemingly in perfect health, were able to so live without a single stigma or stoma—so much is certain. The injection method enabled the writer to test the distention theory already mentioned as being held by a goodly number of ob- servers. By injecting a large amount of a dilute solution into the free peritoneal cavity any degree of distention may be produced. This would serve only, of course, if the endothelium of the abdom- inal wall were the object of study. If it is desired to study the endo- thelium covering the intestinal canal a small amount may be in- jected into the abdominal cavity and the intestine then distended with air. Mice, because of their cheapness and convenient size, were most used in these experiments. For the sake of comparison severa! other animals were injected, namely, several rabbits, rats, and a cat. Larger animals, for obvious reasons, were not experimented upon. After many trials under the most varied conditions, it can be stated that stomata do not occur under these conditions. This method of testing the distention theory would seem to be an ideal one, for the tissues are uninjured in any way, and there is no escape of blood or lymph to obscure the results. On account of the influence the cell form was believed to have on the production of the stomata, it will be proper to consider the effect of simple distention on the cell outline. The distention ex- periments before mentioned show that the cells are large enough to cover the membrana limitans without changing their form to any appreciable extent. When a serous membrane has less surface to cover it forms folds, and the individual cells do not contract. These folds are microscopic, and they have given rise to confusion because various deposits occurring in them have been mistaken for stomata. STIGMATA AND STOMATA OF THE PERITONEUM 73 Too much stress can not be laid on the importance of experimental study on this point on account of its bearing on the problems of stomata formation. Another method of determining the effect of distention may be briefly mentioned. An abdomen may be distended with air and the intercellular spaces made apparent by painting the surface of the abdomen with a solid stick of silver nitrate. The cell outlines are brought into view. No influence on the production of stomata can be observed. It may be mentioned in passing that the silver thus applied acts not only through the whole thickness of the abdominal wall, but to a considerable depth beyond. This is, in a sense, a con- tradiction to the generally accepted opinion that the action of silver is very superficial. Still another way of demonstrating the absence of openings be- tween cells is as follows: A bit of the abdominal wall is stuck to a slide, endothelial surface down.” After firm adhesion had taken place, the membrana limitans with all the other tissue may be removed, leaving the endothelium alone adherent to the slide. It may now be treated with any desired stain, preferably fuchsin. The edges of the cells will be found to lie closely together without the existence of openings. The last method gives some very satisfactory results, though the method is somewhat uncertain.® In the second group may be considered together all those struc- tures that have been regarded as stigmata or stomata which are ex- cluded in the first group. This will include all structures that have been described as such in the literature. Structures identical with those of the first group have been figured in every conceivable place that silver can penetrate. In many cuts the stigmata may be seen scattered promiscuously over the cells.® Stomata likewise may occur in a great variety of places. They may often be seen in the body of the cell forming frequently gyrated figures (see fig. 1, pl. XII). Again the circles may occur in the intercellular line but in which’ the line continues uninterrupted through it. These must be distinguished from objects occurring 7Afanassiew’s method (1. c., p. 58) was used. 8 The writer has for some time been experimenting with iron and tannin for bringing out the cell outlines. By this method, likewise, no stomata appear, nor are the cell outlines af- fected by stretching. 9 Meyer very properly asks that, if they are ‘‘to be called stigmata in one situation, why not in the other?”’ 1.c. Robinson is the only writer who is willing to accept them in this ab- normal situation (The Peritoneum, p. 53. Chicago, 1897). 74 ARTHUR E. HERTZLER below the endothelium. These stomata may be seen below the endo- thelium, between the muscle fibers (fig. 3, pl. XII). These may, instead of forming rings, form solid patches, thus resembling a cell. These were pictured in Oedmasson’s original publication, and they may be seen in many productions since that time, but it re- mained for Meyer to recognize the true conditions. Fig. 10 in his paper presents the appearance very well. These same structures may be seen lying upon the endothelium of the lymph vessels. A modification of the regular stomata, already alluded to, is in- sisted on by a number of investigators: it is the occurrence of open- ings in the center of radiating groups of cells. Nikowlsky laid especial stress on these and pictures several marked examples. A few writers, notably Klein, regard these as the only true stomata. These are certainly striking figures, but it is difficult to understand how these came to be regarded as the true stomata, for it often requires prolonged search before one can be found. Their rarity alone ought to be sufficient to refute the arguments that have been advanced relative to the function ascribed to them. Attempts to study them on cross-section have been fruitless. The significance of the occurrence of extraneous cells between the regular endothelial cells has been the theme for a great deal of discussion. Broadly speaking, they have been regarded as either leucocytes or young endothelial cells, or again as cells destined to form endothelial cells (Keimgellen). No attempt has been made to separate the cells actually occurring between the endothelial cells from those located either above or below them. This is quite nec- essary since these cells have often been regarded as guards for the stomata, filling them up valve-like, as it were (Ranvier), or, when impregnated with silver, they have been described as stomata. Much confusion has arisen because no attempt has been made to determine the exact location of the cells. This is beautifully illus- trated by a cut Robinson’® borrows from Stohr, in which nuclei of cells lying underneath the regular cells are shown. Stohr’ regards them as nuclei of connective tissue cells (see fig. 6, pl. XII), and Robinson calls attention to the fact that these have always been re- garded as stomata. It is quite imperative that this differentiation be carried much farther. This is often not an easy matter, for even 1 l.c., p. 29. U1 Std6hr, Lehrbuch d. Histologie, 7te Aufl, S. 220. STIGMATA AND STOMATA OF THE PERITONEUM 75 with modern microscopical appliances it is often difficult to determine whether a particular object lies above or below the endothelial cells. This is more difficult in silver preparations because the silver is thrown down in fine granules, which cause a confusing refraction of the light. When objects are below the intercellular line it is often possible to determine this fact. With this exception it seems advis- able to exclude all evidence that can not be proved on cross-section. Cells above the endothelial cells are but rarely found. The writer for a long time observed cells in this situation, believing them to be endothelial cells, which were contracted from irritation, as de- scribed by Ranvier. On cross-sectioning the tissue the endothelial layer was found to be intact. Attempts to determine their relation to inflammatory processes, experimentally produced, have been with- out result. The first observer to note the occurrence of small cells between the larger ones was Dybkowsky. Klein advocated the view that they were cells in course of development. Ranvier regarded them as leucocytes filling up the stomata. Virchow’ has been incorrectly quoted as advocating this view. As a matter of fact the statement was made that “multinuclear cells occur below the epithelial cells of the peritoneum.” These words were written before the controversy began and before it was known that leucocytes occurred outside of the blood-vessels ; indeed it was in support of the theory that they do so occur that the statement was made. The leucocytes are really the only ones that ought ever to have been mistaken for stigmata or stomata, for, as Tourneau and Hermann pointed out, the oblong nuclei and the bright nucleoli are always sufficient to distinguish the other cells occurring in this locality from them. This discus- sion could have arisen only from a failure to study them in cross-section. The exact significance of these cells is not patent to the subject; it is sufficient for the purpose to show that they form a part of the regular cell layer. It is the cells located below the endothelial cells that have been mistaken for stomata the most frequently, and in consequence are of greater interest in this connection. Many fine examples of this confusion may be seen in the literature, notably in the publication of Oedmasson. Kolossow classes them all as leucocytes. As al- 12 Gesamte Abhandlungen, S. 167. 1856. 76 ARTHUR E. HERTZLER ready mentioned, Stohr pictures several as connective tissue cells. Both kinds do no doubt occur in this place. There are still others occurring here probably belonging to the clasmatocytes of Ranvier*® or to those large cells which are now regarded as taking some part in the process of inflammation. It 1s sufficient to say that they are all located below the-endothelial layer. It is necessary to note that there are fields without nuclei oc- curring between the endothelial cells. The explanation for this occurrence is not easy. It is quite possible that it is a cell below the regular cells taking its place in the regular layer. With the silver method alone it is quite impossible to distinguish the various cells from one another, and this problem must be solved by the aid of other stains. With these it may be determined to a certainty that the great variety of cells that have been at some time or other believed to bear some relation to the stomata are entirely independent of the endothelial layer. Their exact office is a problem that remains to be solved. The silver method will not aid in solving it. Indeed it is to the uncertainties of this method that much of the confusion is due, aiding much in making the histology of the peritoneum “the darkest chapter in anatomy.” CHEMISTRY Now, thoroughly convinced that the stigmata and stomata are spurious products formed by a precipitate of silver, a determina- tion, if possible, of their exact chemical composition seemed very necessary. As is well known, the intercellular lines are supposed to be formed by the action of the silver nitrate upon the albuminous cement substance, forming an albuminate of silver. This assump- tion has been accepted by all writers. In approaching this subject the writer thought to prepare himself for the investigation by studying the chemical nature of the substance under consideration. He was surprised, after spending some time in a literary search, to find no mention of silver albuminate in the most extensive works on chemistry, in any of its departments. Surprise rose to amazement when one of the ablest of chemists was appealed to for aid and the reply was made that he knew of no such substance. Here, then, was a virgin field for investigation. Returning to the 13‘*Des Clasmatocytes,”’ Archives d' Anatomie Microscopique, Tome III, p. 122. STIGMATA AND STOMATA OF THE PERITONEUM 77 observation of Ranvier that the occurrence of stomata could be pre- vented by rinsing the membrane before applying the silver, it was quite natural to remove some of this fluid for examination. Some of the fluid normally covering the free surface of the peritoneum was carefully transferred to a slide and exposed to the action of a very dilute solution of silver nitrate. Minute droplets of this mix- ture were then transferred to a clean slide and placed under the microscope. What were clear droplets became, on exposure to the sun, dark brown rings. The same thing occurs when any stain is evaporated: what was a droplet becomes on evaporation a ring with a faintly stained center. This is too common an observation to need mention. The rings produced by evaporating small drops of silver and lymph (or whatever the composition of the substance removed may be) bear a striking resemblance to the stomata produced in the regular manner, only of course they are larger. The discovery was accidentally made that, if the two substances were mixed in larger amounts and allowed to evaporate and then examined in glycerine, dots and rings appeared indistinguishable from the true stigmata and stomata. So striking are the pictures so produced that the state- ment seems warranted that stigmata and stomata may be produced independent of a serous membrane. This statement is equivalent to saying that the stigmata and stomata are the product of the action of a solution of silver nitrate upon lymph. The microscopic appearance of silver chloride bears a very strong resemblance to the product of silver and lymph, though the color is different. Dots corresponding to the stigmata may be seen and a few rings, but they are much less constant than in the silver and lymph product. It may be mentioned in passing that the granules seen in the tissues in argyra bear a very strong resemblance to these dots of silver chloride. In order to test the silver chloride theory of the formation of the stigmata and stomata, the following experiment was performed: Five cc. of a sodium chloride solution was injected into the free peritoneal cavity of a mouse. After a few minutes a dilute solution of silver nitrate was injected through the same needle. After the silver solution had had time to mix thoroughly with the sodium chloride solution, the needle was withdrawn, the opening made by the needle ligated, and the animal given the liberty of the cage. Presumably the silver was precipitated as soon as it passed the point 78 ARTHUR E. HERTZLER of the needle, though absolute proof of this is difficult to produce, since the reaction takes place in a perfectly dark chamber, namely the free peritoneal cavity. Attempts were made to obtain an im- pregnation by first mixing the silver nitrate with the sodium chloride solutions and afterwards injecting. The results were negative. A trial was then made of the effect of different colored lights on these solutions made before injection. The results likewise were nega- tive. The remarkable fact was noted that when the silver solution was preceded by a sodium chloride solution the lines appeared at once. When the silver solution was alone injected the lines did not appear until from one-half to three hours afterwards. In either case the appearance of the lines is the same. The difference was noted that when the silver solution was preceded by a sodium chloride solution there always occurred precipitates over the body of the cell, which is not the case when the silver is used alone. An attempt was made to change the reaction of the albuminous sub- stance covering the peritoneum before injecting the silver solution. The means employed to accomplish this were to inject a solution of equal parts of hydrochloric acid (C. P.) and distilled water, fol- lowed after a few minutes by the silver solution. The lines ap- peared. The reader is allowed to judge for himself what effect an acid of this strength would have upon the peritoneal lymph. It was interesting to note that the endothelial cells stood this severe treatment with little injury. It was interesting, too, to note that when so treated the intercellular lines were very fine, with no broad- enings whatever. As an example of the striking figures produced by the action of the silver nitrate solution on an albumin solution figs. I and 2, pl. XIII, may be referred to. The specimen from which this drawing was taken was made by mixing a drop of % per cent of hydro- chloric acid with a dilute albumin solution, and the mixture gently warmed and allowed to evaporate. The peculiar radiating arrange- ment of the rows or rings is due, no doubt, to the properties of the albumin solution. What the exact composition of the product of silver and albumin is can not be stated. The fact that the fluids of the body contain 0.65 per cent of sodium chlorid, taken together with the well-known fact that the nitrate of silver in solution is more sensitive to chlorids in loose combination than to any other substance, has caused the STIGMATA AND STOMATA OF THE PERITONEUM 79 writer to accept as a working hypothesis the assumption that the substance is a chlorid of silver, perhaps holding an albuminous substance as an admixture. This assumption has to support it the following facts: (1) That no such substance as silver albuminate is known; (2) only when a chlorid-containing substance is present are they formed; (3) the sensitiveness of the silver nitrate in solution to chlorids in loose combination; (4) combinations of silver with nuclein are known, but nucleins are not contained in the substance in question. This is but an hypothesis, it is true, and perhaps one of uncertain importance, but the fact remains that it is an unwarranted assump- tion to refer to any substance as silver albuminate. The ultimate solution of this problem may be left to those skilled in chemistry. Indeed, so far as the problems in the histology of the peritoneum are concerned, the exact composition of these substances is a matter of little importance. But it is a problem of vastly greater importance to recognize them as chemical products, the product of the action of silver nitrate in solution upon a chlorid-containing substance. That the stigmata and stomata occur by no other method has already been stated, and the converse is equally true—that by all other methods, without a single exception, the endothelial cells are shown to contain no such openings. Halstead, Kan. 80 ARTHUR E. HERTZLER WORKS CITED #4 1. FLINZER, A. De argenti nitrici usu et effectu in oculorum morbis sonandis. Desert. Leipsisae, 1854. 2. His, W. Beitrage z. norm. u. path. Histologie d. Cornea, Basel, 1856; and also Vir- chow’s Archiv., Bd. 20, S. 207. 3. V. RECKLINGHAUSEN. Die Lymphgefasse und ihre Beziehung zum Bindegewebe. Berlin, 1862. 4. OEDMASSON. Beitrage zur Lehre von dem Epithel. Virchow’s Archiv., Bd. 28, S. 361. 1863. ». His, W. Ueber das Epithel des Lymphgefass-systems und tiber die v. Reckling- hausen’sche Saftcanale. Zettschrift f. wiss. Zoologie, Bd. XII1.,S. 455. 6. LUDWIG UND SCHWEIGGER-SEIDEL. Ueber das Centrum tendenium des Zwerchfells. Arédeiten aus der physiolog. Anstalt zu Leipzig. 1866. . DYBKOWSKY. Ueber Aufsaugen und Absonderung der Pleurawand. Arbeiten aus der physiolog. Anstalt zu Leipzig. 1866. 8. SCHWEIGGER-SEIDEL UND DOGIEL. Ueber die Pleura-hohle beim Frosche und ihren Zusammenhang mit dem Lymphgefdass-system. Beitrage der sach. Gesellschaft d. Wuissen- schaften. 1866. 9. COHNHEIM, J. Ueber Entziindung und Eiterung. Virchow’s Archiv., Bd. 40, S.1. 1876. 10. AUERBACH. Virchow’s Archiv., Bd. 33, S. 381. 11. AFANASSIEW. Ueber den Anfang der Lymphgefdsse in den serosen Hauten. Virchow’s Archiv., Bd. 44, S. 37. 12. Foa. Ueber die Beziehung der Blut- und Lymphge'asse zum Saftkanalsystem. Virchow’s Archiv., Bd. 65, S. 284. 13. KLEIN, E. Anatomy of the Lymphatic System. London, 1875. 14. Kix, E. Grundziige der Histologie, S. 98. 15. KLEIN AND NOBLE SMITH. Atlas of Histology. London, 1880. 16, TOoURNEAU. Recherches sur 1’ anatomie des seruses. /our. de l’ anatomte et de phys., p. 66. 1874. 14The writer believes that he has read, with a very few exceptions, all the original papers that have been written on the subject, and the works here quoted are selected for one of the following reasons: (1) That they were the first to advance a certain theory; (2) that they had unusual influence in forming accepted opinions; or (8) because of their recent date. A. 18, 19. 20. 22. 23. 27. STIGMATA AND STOMATA OF THE PERITONEUM 81 ARNOLD, J. Ueber Diapedesis. Virchow’s Archiv., Bd. 58, S. 203. 1873. ARNOLD, J. Ueber die Beziehung der Blut- und Lymphgefasse zu den Saftkanalen. Virchow’s Archiv., Bd. 74. ARNOLD, J. Ueber die Kittsubstanz der Endothelien. Virchow’s Archiv., Bd. 66, Sil. ARNOLD, J. Ueber die Durchtrittsstellen der Wanderzellen durch entztindete serdse Haute. Virchow’s Archiv., Bd. 74, S. 245. . MUSCATELLO. Ueber den Bau und Aufsaugensvermogen des Peritoneum. Virchow’s Archiv., Bd. 142. RANVIER, L. Lehrbuch d. Histologie; Uebersetzt von Hicati und Wyss. Leipsig, 1888. KoLossow. Ueber die Structur des Pleuro-peritoneal und Gefadssepithels (endothels). Archiv. f. mikroskop. Anatomie, Bd. 42, S. 318. 1893. . Ussow. Le Physiologie Russe. Abstract in Merkel und Bonnet’s Ergebnisse, S. 691. 1899. MEVER, A. W. The Epithelium of the Peritoneal Cavity of the Cat; Contributions from the Anatomical Laboratory of the University of Wisconsin, Bulletin of the University, No. 23. 1900. . KOLossow. Ueber eine neue Methode zur Bearbeitung der Gewebe mit Osmiumsaure. Zeitschrift fiir wissenschaftliche Mikroskopie, Bd. 1X, S. 38. And also, Erganzungsbemerkungen tiber meine Methode der Behandlung der Gewebe mit Osmiumsdaure. Ibid, S. 316. KorTE, W. Article on Peritonitis, in Handbuch der praktischen Chirurgie, Herausge- geben von KE. v. Bergmann, P. v. Bruns und J. v. Mikulicz, Bd. III, Sot ) LOLDT. Lehrbuch der Gewebelehre, III Aufl.,S. 255. 1888, 82 ARTHUR E. HERTZLER EXPLANATION OF PLATES % Plate XII Fig. 1. Mesentery of cat, showing a variety of stomata in the body of the cell. These structures differ from the intercellular stomata only in location. Fig. 2. Mesentery of rabbit: @, numerous stigmata; 4, stomata, one of them half stigma and half stoma, c, these structures here shown are not cells, as has been supposed, but are spurious products, the black center lying above, the circle below, the endothelial cells. Fig 3. Mesentery of rabbit. A stoma between muscle cells below endothel- ial cells. Fig. 4. Mesentery of frog showing a number of intercellular stigmata. Fig. 5. Mesentery of rabbit showing stigmata on cross-section. They are located above the intercellular line. Fig. 6. Mesentery of rabbit showing in the intercellular line a nucleus of a cell below the endothelial cells. Whether this nucleus belongs to the connec- tive tissue cells, or to endothelial cells located below the regular layer, or to clasmatocytes, is uncertain. Plate XIII Fig. 1. Rings formed on a slide from a treatment of dilute albumin solution with silver nitrate solution. The rings in this striking figure are indistinguish- able from those formed on serous membranes in the regular way. Fig. 2. Peculiar tracings made by the same method used in fig. 1. This shows in a striking manner the variable figures the solutions named are capable of producing. Fig. 3. Mesentery of young rabbit treated with silver solution after exuda- tion had been excited with an irritant. Fig. 4. Cross-section of specimen from which fig. 3 was taken. It shows that the heavy black precipitate shown in the preceding figure is located above the regular endothelial cells, and not between them, as has been generally be- lieved from a study on the flat surface alone. The endothelial cells have been purposely torn from the membrana limitans for a short distance and left in po- sition the remainder of the section. In the center of the figure a few fibers be- longing to the basement membrane are still adherent to the cells. 15 All figures except figs. land 2, pl. XIII, are redrawn from pencil sketches made withthe aid of the camera lucida with Leitz Obj. 1/j2, Oc. 4. The exceptions noted were drawn direct with Obj. 6, and Oc. 4. PLATE XII . } ae gtk eee owl ag Sal oe A CONTRIBUTION TO THE SUBTERRANEAN FAUNA OF TEXAS* By CARL JOST ULRICH WITH FIVE PLATES In the present paper I propose to describe some of the Arthropods collected by Dr. C. H. Eigenmann in the caves and springs about San Marcos, Tex. The collections were made with a grant from the American Association for the Advancement of Science. The report made by him to the Association at its New York meeting, describing the locality and general character of the cave fauna, may serve as an introduction to the detailed descriptions of species to follow. “In the early part of September, 1899, I visited San Marcos, Tex., to secure, if possible, some living specimens of the cave salamander occasionally thrown out of the artesian well of the United States Fish Commission. This well taps an underground stream about 190 feet from the surface. No specimens of the salamander Typhlomolge came to the surface during my stay, but I received two living specimens from Supt. J. L. Leary. “Besides the salamander three species of Crustaceans had been secured from this well. These were described preliminarily by Mr. Benedict, Proc. U. S. Nat. Mus., Vol. XVIII. One of these, Palaemonetes antrorum, is very abundant, and many are thrown out from the well each day. The eyes of this species are degenerate far beyond those of the blind Cambarus pellucidus of the Mississippi valley caves. They will be described elsewhere. The second one, Ciralonides texensis, is not nearly so abundant as the first. During my stay of three days I secured several specimens. It can readily be seen in the receiving basin of the well when thrown out. “The third, Crangonyx flagellatus, is much rarer, and no specimen 1Contributions from the Zoological Laboratory of the Indiana University under the direc- tion of C.H. Eigenmann, No. 34. 84 CARL JOST ULRICH was secured during my stay. Instead, however, a single specimen of a related species (Crangonyx Bowersii) was secured. “These are all the species that can readily be seen with the naked eye, when swimming about the receiving basin. A screen of bolt- ing cloth (No. 2) placed over the outlet for a short time secured a number of additional species, viz., the front half of a new species of Caecidotea, two new species of Copepoda, a Cypridopsis, and a Crustacean that defied identification and was later lost, as well as a flat worm. The evidence from the screening is that there is yet a rich subterranean fauna to be obtained from this well. “There is near the well a spring arising evidently from the same source by the side of which the well is insignificant in its yield of water. No blind creatures have been recorded from this spring, and the difficulty in straining its output is much greater than that of straining the well. Through the liberal policy of the Hon. G. M. Bowers and Dr. Hugh M. Smith, of the United States Fish Com- mission, a plankton net is now in use at the San Marcos well, and we may expect other additions to the fauna of the well and the underground stream it taps. “Near San Marcos are two small caves. Ezell’s cave was formerly open to the public and provided with steps and other facilities for entrance. The opening leads into a pit about forty feet deep, with one side, that nearest the entrance, quite perpendicular, but with some projecting rocks. At the bottom of this pit and at the side furthest from the entrance a smaller opening led downward to the water, which was said to be about one hundred feet from the en- trance. The Texas variety of small boy has found amusement in rolling rocks down the entrance, thus smashing the steps and closing the former opening at the bottom of the first series of steps. It was necessary to take a side branch to reach the water. This side branch, for sufficient reasons, I did not take to its end, although my assistants managed to get through to the water, without, however, securing any specimens. I was amply rewarded for not entering the deeper recesses by finding in the twilight of the entrance pit an abundant cave fauna. “Not far from this cave is Beaver cave. This is a winding, twist- ing channel of no great height or width. All the available time was devoted to securing specimens, and the cave was not followed to the end. There is no water except in a pit dug in the cave. A CONTRIBUTION TO THE SUBTERRANEAN FAUNA OF TEXAS 85 “Animals, though few in species, were surprisingly numerous in both these caves. The following species were secured in the well and caves :” 1. A flat worm sp.?—Artesian well. Mollusca . Helicina orbiculata-Say. . Vitrea petrophila Bland, pale var. . Bifidaria contracta Say. ee . Helicodiscus Eigenmannz Pilsbry n. sp. J Ol m 0 bo Crustacea . Cypridopsis vidna obesa Brady and Robertson. } . Cyclops cavernarum n. sp. | . Cyclops Learii n. sp. Artesian well. . Caecidotaea Smithit n. sp. | . Ciralonides texensis Benedict. own Ze Ezell’s Cave. ll. Brackenridgia cavernarum un. sp. and genus Te anes 12. Crangonyx Bowersti n. sp. 13. Palaemonetes antrorum Benedict. 14. Larzal crustacean, unidentified. Myriopoda 15. sp.?—Ezell’s Cave. Beaver Cave. \ Artesian well. Arachnida Ezell’s Cave. 16. Theridium Eigenmann? Banks n. sp. Bee aes Thysanura 17. Degeeria cavernarum Pack. \ Ezell’s Cave. 18. Nicoletia texensis n. sp. ) Beaver Cave. Orthoptera 19. Ceutophilus Palmeri Scudder. aye Beaver Cave. Diptera 20. Larval Chironomus.—Artesian well. Vertebrata 21. Typhlomolge Rathbuni Stejneger.—Artesian well. Crangonyx Bowerstin. sp. Pl. XIV. Specific diagnosis——Eyes absent, no trace of pigment. Upper antennae */,—°/, length of body; first joint of peduncle much larger than second; flagellum three times as long as peduncle, consisting of 26 segments; secondary appendage small and slender, of 2 seg- ments. Peduncle of lower antennae little, if any, longer than pe- duncle of upper; flagellum a little longer than last joint of peduncle, consisting of 8 segments. First and second pairs of legs about equal, the propodos considerably wider than the carpus, palm long, 6 86 CARL JOST ULRICH dactyl closing down between two rows of short, notched spines, about 15 in a row. Peraeopoda subequal; first two pairs nearly equal; the third, fourth, and fifth pairs increasing in length back- ward, and with expanded femora. The three pairs of pleopoda well-developed, nearly equal in length, two-branched. First pair of uropoda large and broad, with 2 well-developed branches, second pair similar, but shorter and smaller; third pair very small, inner branch rudimentary. Telson narrow, not emarginate, twice the length of third uropods, with 2 sets of 5 stout setae on posterior margin. Color—White. Length—About Io mm. Hab.—Obtained from artesian well of United States Fish Com- mission at San Marcos, Tex. (Dr. C. H. Eigenmann). This species differs from C. flagellatus Benedict, also obtained from this well, in the number of segments of the upper antennae, being 40-61 in the latter and 26 in C. Bowersti. The flagella of the lower antennae of C. flagellatus consist of 8-19 segments, those of C. Bowersti of 8. The number of the spines on the propodas is also different, there being 15 in C. Bowersu and 24 in C. flagellatus. It differs from C. mucronatus Forbes, in the shape and size of the telson, which, in the females of the latter, is very long and slender; in the males, short and emarginate. In C. Bowersi the telson is about twice as long as broad, and somewhat rounded at posterior margin. The uropods are different, the second and first pair in C. mucronatus being little longer than third, while in C. Bowersiu they are much longer. The flagella of the lower antennae are not pro- vided with olfactory clubs. Additional details—Having but a single specimen, it has not been possible for me to describe all the parts as satisfactorily as ! would wish. Body (fig. 1).—The body is compressed, narrow. Segments of peraeon increase in size from first to fourth. The fourth, fifth, and sixth are nearly equal. Inferior margins rounded. The first, sec- ond, and third segments of the pleon are large, exceeding the seg- ments of the peraeon in length. The fourth, fifth, and sixth segments decrease in size backward. The last segment (telson, fig. I1) is as long as fifth and sixth combined, and narrow. Surface of body smooth. A CONTRIBUTION TO THE SUBTERRANEAN FAUNA OF TEXAS 87 The upper antennae (figs. 1, 2, 3) are about 34 the length of the body. Peduncle of three segments: first broad and stout; sec- ond half as thick as first, not quite as long; third, about */, length of second, somewhat smaller. A few scattered spines on segments of peduncle. Primary flagellum three times as long as peduncle, consisting of 26 segments, all provided with spines, and all, except the first seven or eight and the last provided with a slender olfactory club. Secondary flagellum of 2 small segments, the first about four times as long as the second, and together a little longer than first segment of primary flagellum; provided with spines. The lower antennae (figs. 1 and 4) are*/, as long as upper. The peduncle consists of 5 segments: the first, second, and third short and thick, the fourth and fifth much longer, less thick and spiny. The flagellum consists of 8 segments, each with a few spines, but no olfactory clubs, and together a little longer than third segment of peduncle. Peduncle of lower antennae is not quite as long as that of the upper, but, being inserted somewhat forward of the latter, it reaches as far as peduncle of upper antennae. The propodos (figs. 6 and 7) are about equal in length, which is 2 mm. The hand of the first is large, with a large palmar surface. Dactyl closes down between two rows of notched spines, a hair growing from the notch in each. There are about 15 such spines in each row. The carpus is about % width of hand, nearly triangular, the short posterior margin provided with a row of spines. The coxa has 4 or 5 long slender spines on its posterior margin. The second pair is smaller, the hand being +/, wider than carpus, the latter larger than in first and more rounded, the posterior margin with three transverse rows of 5 spines each, as in C. mucronatus. The peraeopoda (figs. 8 and 9g) are much alike in structure. The first two pairs are rather short and do not have their femora ex- panded as is the case with the remaining three pairs. They are all more or less spiny and end ina claw. Fig. 8 shows a gill-like struc- ture attached to the proximal end. Length of seventh leg, 4.5 mm. ; width of femur, 34 mm. The pleopoda (fig. 10) are well developed. The basal portion is about 34 mm. in length and rather stout. The branches are well developed, a little longer than basal portion and fringed with long plumose setae. - The uropoda (figs. 1 and 11). The first pair is very large. The 88 CARL JOST ULRICH basal portion is equal in length to the third and fourth abdominz segments, while its two branches are nearly as long. The latter ex tend beyond the telson and the second and third pair of uropod: They end in 4 large spines. The second pair is much like the firs in structure, but shorter and smaller. The third pair has a shor basal portion, and one branch, the outer ending in 2 spines. Th inner branch is an unarmed rudiment. In length the third uropo extends but a little beyond the peduncle of the second pair. I take pleasure in naming this species for the Hon. G. M. Bower: United States Commissioner of Fish and Fisheries. Ciralonides texensis Benedict. Proc. U. S. Natl. Mus., p. 615, 189: PEW: The body of the largest specimen examined was 16.5 mm. i length with a width of 6.5 mm. In shape it is oblong elliptical. Th depth is greatest at about the middle of the body, being about +, the breadth. All of the segments have the pleura produced be low and posteriorly into scale-like projections, thus protecting th insertion of the limbs. Above, the surface is smooth. The head is short, convex toward the top and front, and, see from above, almost circular in outline. There is no trace of eyes. The first segment of the peraeon is about twice as long as th second. It widens a little inferiorly, the antero-inferior parts partl surrounding the head. The posterior margin is nearly straigh The inferior margin is slightly convex. The second segment is th shortest of the seven composing the peraeon. The third, fourth fifth, sixth, and seventh are much the same in size and shape. The are produced posterio-inferiorly, and this becomes more pronounce toward the pleon. The segments of the pleon are much shortened, and all five c them together are little longer than one segment of the peraeor They, too, are produced into a point inferiorly, but do not exten down quite as far as the segments of the peraeon. The first seg ment of the pleon is almost hidden by the last of the peraeon. A in other representatives of this family, the pleon is very distinc from the peraeon. The telson is large and well-rounded behind. In width it some what exceeds the pleon. The antennae consist of (a) a basal portion, composed of 5 seg ments, the first broad and short, the second and third about equal i length to the first and */; as wide, the fourth twice as long as th A CONTRIBUTION TO THE SUBTERRANEAN FAUNA OF TEXAS 89 third, and of nearly equal width, and the fifth */, longer than the fourth and */, its width; and (b) a flagellum consisting of 33-35 segments, of which the first is the longest, the second and third about as long as broad. The remaining segments are relatively longer and taper gradually toa point. There are setae at the joints, and the last segment terminates in a number of hair-like bristles. When applied to the sides of the body the antennae reach to the ‘anterior margin of the seventh segment of the peraeon. The antennulae have a basal portion consisting of three articles of which the first is nearly spherical; the second and third are elongated, about three times as long as broad. The flagellum con- sists of 15-16 segments, the first very short, the second longest, and the remaining ones all longer than broad. There are a few setae at some of the joints. The antennulae are about */, the length of the antennae. The first pair of legs are short and stout, armed with a strong claw at the end and a number of spines along the inner margin. From their position and structure, it might be inferred that they are used in grasping food and conveying it to the mouth. The remaining four pairs of legs are longer and more slender, serving as organs of locomotion. In structure they are much alike. Each is armed along the inner margin with spines and ends in a double spine forming a sort of claw. The pleopoda are small and are completely covered by the pleon. Each consists of a short, basal portion to which are articulated two distal branches. The larger, outer branch consists of a broad, thin lamella, unsegmented in the first pair, but distinctly segmented in the remaining four pairs. Of these segments, the first is the larger, rather broad, almost square, while the second is shorter, rounded at distal margin, and fringed with a row of plumose bristles. This outer branch seems to be of firmer texture, and covers the more delicate inner branch. The inner branch in the first and second pairs is nearly as long as the outer, but much narrower, and con- sists of a single segment, fringed with plumose bristles at the ex- tremity. In the remaining pairs this inner branch is also seg- mented, a fact which seems quite out of the ordinary in Isopods. The first segment is short and triangular, the second broad and rounded, and not fringed with bristles. In Bronn, Klassen und Ordnungen des Thier-Reichs, Vol. V, go CARL JOST ULRICH 2, Pp. 35-36, a comparison is made between the pleopoda of the Isopods and the so-called pedes fissi of Copepods. In the former the rule is that the branches are not segmented; in the latter that they are segmented, and the usual number of segments is 3. But there are Copepods in which the number of segments in both branches is reduced to 2, and even some in which the inner branch consists of a single segment. And, on the other hand, there are Isopods in which the outer branch consists of 2 segments, so that we notice a tendency of the two orders to approach each other in this respect. “Unter allen Umstanden bekunden schon die vorste- hend aufgefiihrten die an den Isopoden-Spaltbeinen hervortretende Tendenz, den Aussenast eine Gliederung eingehen zu lassen, wahrend der innere eine solche stets vermissen lasst, zur Gentige.” It will be noticed that the last statement in this quotation does not hold true in the present case. Nevertheless, the fact that in this case both branches are segmented adds another proof to the above statement, that the two orders approach each other. The uropods are considerably shorter in this species than in other Isopods, extending but a little beyond the telson. They are stoutly built, with a triangular basal portion nearly as long as the distal portion. Of the latter, the inner branch is three times as wide as the outer, and its inner margin, as well as that of the basal por- tion, is fringed with plumose bristles. The extremities of both branches are provided with a number of stiff bristles. The mouth-parts indicate that the animal is not parasitic, being clearly adapted for biting. The mandibles are large and strong, somewhat unequal, with a hard, rough cutting surface which is almost black in color. The maxillae are provided with several long, pointed teeth, their dark color indicating their hardness. The maxillipeds and palpi are much the same as in other related species, and are perhaps sufficiently described by the drawings. Brackenridgia Eigenmann and Ulrich, n. gen. Pl. XVI. Eyes none. Antennulae absent. Peduncle of antennae of 5 segments, flagellum shorter than fourth or fifth segment of peduncle, bristly. Legs long and slender, all ambulatory, increasing in length posteriorly. Pleopods with air-cells. Outer branch of uropods longer than pleon, conical. Inner branch much smaller, spiny. Mouth-parts much like those of Titanethes Schioedte; right man- dible with two appendages back of cutting surface, first short, sec- A CONTRIBUTION TO THE SUBTERRANEAN FAUNA OF TEXAS QI ond twice as long, fringed on inner side; another fringed appendage on the hind cutting surface. Left mandible with two fringed ap- pendages next to cutting surface. Third maxilliped (labrum) with 3-segmented palpus, and two small projections on anterior margin. Body somewhat arched, oval, epimera drawn out postero-inferiorly. Last segment of pleon bluntly triangular. Cave dwellers. Allied to genus Titanethes Schioedte, which occurs in caves in Europe. This genus has been named in honor of Mr. G. W. Brackenridge, of San Antonio, Tex., a liberal patron of the natural sciences. Brackenridgia cavernarum n. sp. Pl. XVI, 1-0. This interesting species, which forms the type of a new genus of the Isopoda, was collected by Dr. C. H. Eigenmann during the sum- mer of 1899 in two caves near San Marcos, Tex. These caves are known as Ezell’s and Beaver caves, and are but a short distance apart. The animal seems to be quite abundant here, some twenty or thirty having been obtained in a short time. Up to the present time no representative of the European genus Titanethes or of any closely allied genera has been described for the United States, and this genus and species form an interesting addition to the crustacean fauna of this country. In size this species is rather small, being from 2-6 mm. in length. The body is slightly arched. The head is round in front, its pos- terior margin quite straight. The first thoracic segment partly surrounds the head on the sides. The thoracic segments are nearly equal. The inferior margins are drawn out to a point posteriorly, this being emphasized toward the pleon. The lower margins of the segments are beset with minute spines. The pleon is short, the segments not easily recognized, and the inferior margins not con- spicuously drawn out. The sixth abdominal segment is slightly longer than the others, and bluntly triangular at posterior margin. The latter is beset with short spines. There seems to be no trace of eyes or antennulae. The antennae are about 4 the length of the body, and consist of a basal portion of 5 segments and a short flagellum of 8 segments. Of the basal portion, the first three segments are short and stout, with a few short spines. The fourth and fifth segments are much longer, the former being 2% times the length of the third, with equal width, the latter about 11/, times the fourth, with nearly equal thickness. Both are beset with short spines along the sides, and the fifth has g2 CARL JOST ULRICH one specially long spine near its distal end. The flagellum consists of 8 segments, rapidly tapering to the small extremity, and alto- gether less than the fifth basal segment in length. Each segment has a row of stiff hairs, probably sense organs of some kind, around its middle region. The last segment ends in a number of bristles. The mouth-parts are such as we would expect in a non-parasitic Isopod. The outer maxilliped (labrum) consists of a broad blade with two pointed appendages in front which extend slightly be- yond the head, and a three-segmented palpus. The first maxilla consists of a narrow blade fringed with bristles on the outer side and provided with a cutting surface having five notches, two being hard and three of softer texture. The inner branch of the same is also rather slender, with three teeth, two of them rounded and delicate, one longer and fringed. The mandibles are large and strong. The inner margin of the right mandible is provided with three appendages, the first short, the second and third longer and fringed along the inner margin. On the left mandible we find two equal fringed appendages, corresponding to the first and second in the right mandible. The legs, all of which are ambulatory in character, are much alike in structure, consisting of five segments and a claw composed of two more. Short spines with broad bases are found scattered all over the surface of the legs, together with some longer ones, especially on the last segment. The claw needs perhaps a little more description. It consists of 2 segments, the first, larger one, ending in a spine on the concave side, and a shorter, pointed end segment. Besides these there are two appendages, one on the convex side as long as the claw and rather thin and narrow, ending squarely, and a pointed one, not quite so long and wide, on the concave side. In the allied genus Titanethes Schioedte these appendages end in a tuft of hairs, but nothing of the kind could be made out here. The legs increase in length toward behind, the last being half the length of the body. The pleopoda are very small. As in other terrestrial Isopods, the anterior ones contain air-cells. The uropoda consist of a broad basal portion with a stout conical outer branch, longer than the whole of the pleon, and tipped with four or five bristles, the longest of which are about 34 the length of this branch. The inner branch is much smaller and shorter, beset A CONTRIBUTION TO THE SUBTERRANEAN FAUNA OF TEXAS 93 with numerous short spines on the inner margin, five or six, slightly longer, on the outer margin, and a stout spine at the tip. The basal portion of the uropods projects beyond the posterior margin of the telson on the outer side, but is covered on the inner side. Color—White; a dark longitudinal band often seen along the median dorsal surface, caused by the dark contents of the alimentary tract. ; Caecidotea Smithiin. sp. Pl. XVI, figs. 10-18. Body of loosely jointed segments. Head as in C. stygia Pack. No trace of eyes. Inner antennae short, not more than half as long as basal portion of outer antennae. Flagellum of inner antennae consists of five segments, the second % of first, remaining ones longer. Last segment of flagellum with a spine more than twice length of segment, beside which there is an olfactory club ?/, as long. Another somewhat shorter olfactory club on penultimate segment. Last segment of the basal portion of the inner antennae provided with three spines, as in C. stygia. Outer antennae probably as long as body. Basal portion of 5 segments, the first three short and thick, the fourth and fifth much longer and more slender. The flagellum consists of at least 40 segments. Mouth-parts essentially as those of C. stygia. Legs long and slender, first pair sub-chelate, remaining ones with a weak claw. Inferior margin of the body seg- ments beset with short spines. - Size—Very small, probably not over 3 mm. in length. Color.—White. Hab.—Subterranean stream near San Marcos, Tex. Collected by Dr. C. H. Eigenmann from the United States Fish Commission well The above description is from a fragment. The telson and caudal appendages were gone, also part of the outer antennae. The writer hopes soon to receive the material which will enable him to fill out the gaps in the above diagnosis. In honor of Dr. H. M. Smith, in charge of scientific inquiry of the U. S. Fish Commission. Palaemonetes antrorum Benedict. Proc. U. S. N. Mus., p. 615. £895. Pl: XVIT. Prothorax continued forward into a short, sharp rostrum, the upper margin of which is notched, there being about 12 notches, with plumose spines between the notches, also a few of these on the under side of the rostrum. Just below and to the outside are the 94 CARL JOST ULRICH eye-stalks with very degenerate eyes. There is but one little group of cells left which indicates the original structure, but as far as practical use is concerned the eyes must be entirely functionless. Below the eye-stalks are the inner antennae, or antennulae, and be- low and to the outside of these the outer or true antennae. The former consist of a stalk composed of 4 segments, the first and sec- ond of which are broad and on the under side are continued into a keel-like structure fringed with plumose bristles. The third seg- ment is narrower and a little longer than the others, while the fourth is very short. This bears three appendages, the outer one being the longest, equalling or slightly exceeding the length of the body. The inner flagellum is little over half the length of the outer. The third appendage is short and pointed, and bears along its inner margin a row of structures resembling olfactory clubs. The other two appendages have about the same structure as the true antennae, consisting of numerous segments which gradually taper off to the end. The outer antennae spring from a broad base consisting of three segments. The second of these divides, one part being con- tinued into a large and broad antennal scale fringed with long bristles, and extending beyond the rostrum one-half its length. The third basal segment is much smaller than the others, and much less in thickness. The flagellum of the antenna consists of numerous seg- ments, gradually tapering to the extremity. The total length of the antennae is 26-27 mm. in specimens whose body length is 17 mm. The first and second pairs of maxillipeds (figs. 13 and 12) are somewhat alike in structure, consisting of a broad, flat portion, stronger in the second, provided with numerous spines and bristles. The third maxilliped has much the same character as a foot (fig. 11). All three are provided with a slender palpus, which ends in a tuft of hairs. The legs are all very long and slender. The first and second pairs are chelate, but the chelae are so small that they can be of but little use to the creature. The remaining three pairs end in a crooked claw. The swimmerets are well developed. There is a stout basal por- tion and two branches, an outer longer and stouter one, and an inner shorter one. The latter undergoes interesting modifications in the several pairs. In the first it is short and broad, membranous, and fringed with scattered bristles. In the second it has two little A CONTRIBUTION TO THE SUBTERRANEAN FAUNA OF TEXAS 95 branches springing from the inner side, the first unarmed, the second with a number of bristles (not plumose). In the third there is but one unarmed side branch, and the same is true of the fourth and fifth. In the basal portion of the fourth we find a single spine on the inner margin, but in the fifth both inner and outer margins are fringed with scattered bristles. The two distal branches are closely fringed with long plumose bristles. The telson consists of a conical central portion, its posterior mar- gin beset with two larger and several smaller bristles, and the broad side portions, modified swimmerets, all except the outer margin of which closely fringed with plumose bristles. The gills are completely covered by the carapace. They agree in structure, etc., with those of the allied families of crustaceans. Cyc.ops: Two specimens of this widely distributed genus were found in the water of the artesian well mentioned above. They are both blind, as far as external examination can show. They were pre- served in formalin which, in other cases at least, did not decolorize the eyes if there were any. The following descriptions are in- tended to be preliminary only, and as soon as the necessary material is at hand a more detailed study and comparison with other related forms will be made. Cyclops cavernarumn. sp. Pl. XV, fig. 18. Antennae 17-jointed, reaching to middle of second thoracic seg- ment. No trace of eye. Cephalothorax oval-elongate. Abdomen stout. Cyclops Learitin. sp. Pl. XV, fig. 19. Antennae 12-jointed, scarcely reaching the posterior margin of first thoracic segment. Cephalothorax oval, shorter than in C. caver- narum. No trace of eye. Abdomen rather slender. Cypridopsis vidua obesa Brady and Robertson. This species was represented by numerous specimens in the col- lections from the artesian well. ER - 96 CARL JOST ULRICH Nicoletia? texensis n. sp. Pl. XVIII. Two specimens were found by Dr. Eigenmann in Ezell’s cave, near San Marcos, Tex. The body of the larger is about 18 mm. in length ; that of the smaller about half as much. The antennae were not complete, but seem to be at least as long as the body. The same is true of the caudal appendages. In form and appearance the body is much like that of Campodea staphylinus Westw., though the head is relatively smaller and the thoracic region larger (see his. 1): Head.—Rather small, rounded on the sides, slightly pointed an- teriorly, convex above (fig. 1). Antennae very long, of numerous segments (in the imperfect specimens I examined I counted about 40, and there might have been as many more, judging from the thickness of the last segment). The basal portion consists of 2 segments, the first half as long as head, rather stout, the next short, as long as broad; first segment of flagellum three times as long as second ; second and following segments short, gradually lengthening toward distal portion. Basal portion as well as flagellum with numerous spines, some of them forked at tip. Besides these, the whole surface is covered with minute hairs (fig. 8). There seems to be no trace of eyes. The labrum is provided with a transverse row of bristles, as well as with some scattered ones. The mandibles (fig. 7) are large and prominent; there are four larger teeth and several smaller ones; the inner grinding surface is provided with a fine, comb-like arrangement. The maxillae (fig. 4) are also large and conspicuous. There is one large, hard tooth, then one with a comb-like arrangement, then follow five with one side finely ser- rate, another with smooth margins, and then a row of stout bristles forked at the tip. The maxillary palp is long and slender, having the appearance of an antenna. It consists of 5 segments, the first short and thick, the remaining ones long and slender ; the second and 2 Key to the genera of Lepismidae by Nathan Banks. 1. With scales Body slender, cerci longer than body, No scales.. 5 Troglodromicus DINO CV ES er osaseeuis claw aeioe sinatieclsinnleie s Geule 4. Maxillary palpi, 5-jointed......... Lepisma With eyes, body slender, cerci longer Maxillary palpi, 6-jointed...... Thermobia thanlabGoneye eee enesatr bee eeee £ D-H Yes (present joan raletslocientnee Maindronia 3. Body broad, cerci shorter than body, yes a DSent soo. .\eremenienee setae ee Nicoletta Lepismina Nicoletia.—Abdominal appendages on segments 2-9, cerci nearly as long as body, no eyes, no scales, maxillary palpi 5-jointed, mandibles tridentate at tip, body not very slender. Four species are known: JV. phytophila Gerv., N. geophila Gerv., N. cavicola Joseph, NV. nagg? Grassi. A CONTRIBUTION TO THE SUBTERRANEAN FAUNA OF TEXAS 97 third each have a row of spines near the distal ends (fig. 5). On the end of the last segment there are five club-shaped structures, probably sense organs of some sort (fig. 6). The whole palp is covered with minute hairs and with larger spines. The labium is broad posteriorly, cleft anteriorly, with large palps, of four joints each, the last broad and heart-shaped, with three short cylindrical structures on the end (fig. 3). The first thoracic segment is little longer than the head, but the second and third are much larger. The margin is somewhat pro- duced postero-inferiorly. The legs are rather long and slender, hairy, with a triple claw (fig. 2). The latter seems hard and is yellow in color. There are 10 abdominal segments. The last two are rather short. The abdominal legs are found on the second to eighth segments. The ninth is provided with two long appendages, while the last ends in one long median appendage. These three have been mentioned above under caudal appendages. The abdominal legs are rather weak, of one joint, ending in a straight claw, with several spines. The caudal appendages consist of numerous segments, lengthening toward the tip, and elaborately fitted out with spines and bristles, some of which are stout and forked, others very long and slender. They seem to be arranged in a definite way (see fig. 9). Color of specimens preserved in formalin: yellowish white. Degeeria cavernarum Pack. Pl. XVIII, figs. 11-13. Two specimens of this common Podurid were obtained from Ezell’s cave. They differ from the form found in the vicinity of Bloomington by the longer antennae. Especially the fourth seg- ment is much longer than in forms found here. DESCRIPTION OF A NEW CAVE SPIDER BY NATHAN BANKS Theridium Eigenmanni n. sp. Female, length ceph. 2.1 mm.; femur I=8.2 mm.; tibia I=7 mm. Cephalothorax, legs, and sternum pale reddish yellow; mandibles a little darker; abdomen variable, sometimes entirely pale, some- times black all over except two pale spots on highest part and a row of two or three reaching to the spinnerets; between these extremes are many grades of markings, some with a few black spots above, others with a row of chevrons behind and several large spots on each side above. 98 CARL JOST ULRICH General structure similar to T. tepidariorum. Cephalothorax rather broad, depressed around dorsal groove; caput rather low, broad; posterior eye-row nearly straight; P. M. E. about twice their diameter apart, a little closer to the sub-equal P. S. E.; A. M. E. about once their diameter apart, a trifle farther from the slightly smaller A. S. E.; M. E. make a quadrangle slightly higher than broad, and broader above than below; mandibles of moderate size; sternum blunt-pointed between the hind coxae; legs long and slender, clothed with rows of fine short hairs; femur I full as long as the cephalothorax and abdomen taken together; tibia I a little longer than metatarsus I; abdomen high, strongly arched above at middle, suddenly descending behind, higher than in most of the allied forms; the lung-plates each side at base are reddish and hardened. The epigynum shows two dark spots with a narrow transverse opening behind. Male, unknown. Specimens from Beaver cave and Ezell’s cave, near San Marcos, Tex.; collected by Prof. C. H. Eigenmann, after whom the species is named. The specimens from Beaver cave are darker than the others. 10. 11. A CONTRIBUTION TO THE SUBTERRANEAN FAUNA OF TEXAS 99 CHIEF WORKS CONSULTED . BENEDICT, J. E. Preliminary description of a new genus and three new species of Crusta- ceans from an artesian well at San Marcos, Tex. Proc. U.S. Nai. Mus., Vol. XVIII. 1895. . BLATCHLEY, W. S. Indiana caves and their fauna. a2rst An. Rept. of Dept. of Geol. of Ind. 1896. . BRONN. Klassen und Ordnungen des Thier-Reichs. Arthropoda, Vol. V, Pt. 2. 1892. . CALL, R. ELLSwWoRTH. Sonie notes on the flora and fauna of Mammoth Cave, Ky. Reprint from Am. Nat., pp. 377-92. May, 1897. . CHILTON, CHARLES. The subterranean Crustacea of New Zealand, with some general remarks on the fauna of caves and wells. 1894. . FORBES, ERNEST B. A contribution to a knowledge of North American fresh-water Cyclopidae Bull, Ill. St. Lab. of Nat. Hist., Vol. V. 1897. . FORBES, S. A. List of Illinois Crustacea. ull. ll. Mus. of Nat. Hist., No.1. Dec., 1876. . HAMANN, OTTo. Europaische Hohlenfauna. 1896. HERRICK) Cy T;: Copepoda, Cladocera, and Ostracoda of Minnesota. Geol. and Nat. Hitst. Surv. of Minn. 1895. LEUNIS, J. Synopsis der Thierkunde, Vol. 2. 1886. LUBBOCK, JOHN. Monograph of the Collembola and Thysanura. 1873. POPAGKAR TH. ALCS: The cave fauna of North America, with remarks on the anatomy of the brain and origin of the blind species. 1886. 100 CARL JOST ULRICH EXPLANATION OF PLATES Plate XIV Crangonyx Bowersii Ulrich. . Last three segments of upper antenna; 3a, olfactory club. Fig. 1. C. Bowersii. Fig. 2. Upper antennae, basal portion; 2a, secondary flagellum. Fig. 3 Fig. 4. Lower antenna. Fig. 5. Maxilliped. Figs. 6, 7. First and second gnathopods. Fig. 8. One of the fourth pair of legs; 8a, gill. Fig. 9. One of the sixth pair of legs. Fig. 10. One of the pleopoda. Fig. 11. The telson, uropoda, etc; 11a, fifth abdominal segment; 110, sixth abdominal segment; llc, telson, 11d, first uropod, 1le, second uropod; 11/, third uropod. Plate XV Figs. 1-17. Benedict. Figs. 2-6. Thoracic legs. Figs. 7, 8. Upper and lower antenna. Fig. 9. End of lower antenna. Fig. 10. Uropod. Fig. 11. Mandibles. Ciralonides texensis Fig. 12. Labrum. Fig. 13. Maxilla. Fig. 14. Ventral view of head. Figs. 15, 16, 17. The first, second, and third pleopoda. Fig. 18. Cyclops cavernarum Ulrich. Fig. 19. Cyclops Learii Ulrich. Plate XVI Figs. 1-9. Brackenridgia cavernarum Ulrich. Fig. 2. Antenna. Fig. 3. Mandibles. Fig. 4. Labrum. Fig. 5. Maxilla. Fig. 6. Thoracic leg. Fig. 7. Claw. Fig. 8. Telson, with uropods. Fig. 9. One of the uropods. Figs.10-18. Caecidotaea Smithii Ulrich. Fig. 11. Portion of lower antenna; lla, basal portion of same. Fig. 12. Upper antenna. Fig. 13. Basal segment of upper an- tenna showing auditory spines, an. Fig. 14. Endsegments of upper anten- nae, showing olfactory clubs, o/. Fig. 15. Labrum. Fig. 16. Maxilla. Figs. 17, 18. First and second legs. Plate XVII Palaemonetes antrorum Benedict. Fig..1. P. antrorum. Figs. 2-6. Thoracic legs. Fig. 7. Basal portion of lower antenna. Fig. 8. Portion of third (shortest) flagellum of upper antenna. o/, olfactory clubs. Fig. 9. Basal portion of upper antenna.. Fig. 10. Telson. Figs. 11, 12, 13. Maxillipeds. Fig. 14. Gill. Fig. 15. Rostrum. Figs. 16-20. Pleopoda. PLATE XIV pte eo 5 SF OD Die a / [Thr <<“ S PLATE XV CTIu ~ PLATE XVI aN Ne Wa y ae Ti NK - +70 yo PLATE XVII PLATE XVIII A CONTRIBUTION TO THE SUBTERRANEAN FAUNA OF TEXAS IOI Plate XVII Figs. 1-10, Nicoletia texensis Ulrich. Fig. 2. Thoracic leg. Fig. 3. Labium. Fig. 4. Left maxilla. Fig. 5. Maxillary palpus. Fig. 6. End of max. palpus showing peculiar sense (?)organs. Fig. 7. Left mandible. Fig. 8. Basal portion of antenna. Fig. 9. Two segments of cercopod. Fig. 10. Abdominal leg. Figs. 11-13. Degeeria cavernarum Pack. Fig. 12. End of elator. Fig. 13. Claw. ON THE AMOUNT OF OXYGEN AND CARBONIC ACID DISSOLVED IN NATURAL WATERS AND THE BEFECT OF, THESE) GASES.- UPON) THE OCCURRENCE OF MICROSCOPIC ORGANISMS By GEORGE C. WHIPPLE anp HORATIO N. PARKER WITH FOUR PLATES INTRODUCTION Of the many factors which affect the growth of microscopic or- ganisms in water the most important is their food supply, and of the elements which enter into their composition oxygen and carbon are of fundamental importance. Ordinarily these two elements are not determined in the sanitary analysis of water, and data on their occurrence in natural waters are not as numerous as one could wish ; and especially there are lacking parallel observations upon the amount of carbonic acid and the number of the microscopic organ- isms. With nitrogen the case is different. This element plays a conspicuous part in sanitary water-analysis, and the relation be- tween the amount of nitrogen as it exists in various states and the microscopic organisms present in the water has been care- fully worked out. Nitrogen is usually determined in four states: First, as “albumin- oid ammonia,” that is, nitrogen existing as organic matter; second, as “free ammonia,’ which represents the preliminary stage of de- composition of the organic matter; third, as “nitrites,” which repre- sent a further stage in decomposition; and, fourth, as “nitrates,” which represent the final stage of decomposition, that of complete mineralization. Nitrogen exists also as a gas dissolved in the water, but as such it is seldom determined. It seems to have been taken for granted, and apparently with good reason, that dissolved nitrogen is as inert in its relations with organic life as is the nitrogen of the atmosphere: but it has been discovered that some forms of IO4 GEORGE C. WHIPPLE AND HORATIO N. PARKER terrestrial plant life, aided by the action of certain bacteria, are able to utilize atmospheric nitrogen, and it may be that aquatic plants can in like manner assimilate dissolved nitrogen. The relations that exist between algae growths and the nitrogen contents of water as ordinarily determined are illustrated by Plate XIX. SOLUBILITY OF OXYGEN AND CARBONIC ACID Great variations exist in the solubility of gases in water. At 20° C. one liter of water has the power of absorbing 28.38 c.c. of oxygen, 14.03 c.c. of nitrogen, 901.4 ¢c.c. of carbonic acid, and 654,000 c.c. of ammonia. The coefficient of absorption of a gas is the volume of that gas—expressed in cubic centimeters and measured at 0°C. under a pressure of 760 mm. of mercury—which is dissolved in I c.c. of water. Except in the case of hydrogen, the coefficient varies with the temperature, decreasing as the temperature increases. The coefficients of absorption at various temperatures for oxygen, nitrogen, and carbonic acid are given in the following table: TABLE I COEFFICIENTS OF ABSORPTION OF VARIOUS GASES, AFTER BUNSEN! NVeeues| OXYGEN NITROGEN CARBONIC ACID 0 0.04114 0.02035 1.7967 1 0.04007 .017981 1.7207 2 .03907 .01932 1.6481 3 .03810 .01884 1.5787 4 .03717 .01838 1.5126 5 .03628 .O1794 1.4497 6 .03544 .01752 1.3901 7 .03465 .01713 1.3339 8 .03389 .01675 1.2809 9 .03317 .01640 1.2311 10 .03250 .O1607 1.1847 11 .03189 .01577 1.1416 12 .03133 .01549 1.1018 18 .03802 .01523 1.0653 14 .03034 .01500 1.0321 15 .02989 .01478 1.0020 16 .02949 .01458 .9753 17 .02914 .01441 .9519 18 .02884 .01426 .9318 19 .02858 .01413 .9150 20 .02838 .01403 .9014 1Gasometrische Methoden, Braunschweig. 1877. EFFECT OF GASES ON MICROSCOPIC ORGANISMS 105 The volume of the gas dissolved is the same, whatever the pres- sure, but inasmuch as the density of the gas increases directly with the pressure, it follows that the actual quantity of the gas dissolved varies directly with the pressure. In the case of a mixture of the gases, the quantity of any one of them which will be dissolved de- pends upon the vapor pressure of that particular gas, regardless of the others. Thus, water at 0° C. will dissolve 41.14 c.c. per liter of oxygen if exposed to an atmosphere of pure oxygen under 760 mm. pressure, but if exposed to the air, which is but one-fifth oxygen, it will absorb 41.14 c.c. of oxygen under one-fifth the pressure ; conse- ‘quently only one-fifth as much by weight as in the first case. Again, water at 0° will dissolve 901.4 c.c. per liter of carbonic acid in an atmosphere of that gas under 760 mm. pressure, while in the air of a room where the amount of carbonic acid is only 0.2 per cent, the amount dissolved will be only one-five-hundredth of that quantity by weight. This law is said to be only approximately true in the case of gases which form but a small per cent of any mixture of gases. The coefficient of absorption of oxygen is greater than that of nitrogen ; consequently when water is exposed to air it will absorb a larger amount of the former gas. Bunsen has given the fol- lowing figures to illustrate this: COMPOSITION OF DISSOLVED AIR BY VOLUME COMPOSITION OF AIR BY VOLUME MUKA ON fetes iate sia) s'aic le elias % oils sie teva! Biehelstiecs Sia) epslere 20.96 per cent | 34.91 per cent WON EE SEDs MRC Oa Sonne Been aise aq bass cae 79.04 per cent | 65.09 per cent 100.00 per cent |100.00 per cent It is possible for water to temporarily contain a larger amount of gas than it will actually dissolve under definite conditions of tem- perature and pressure; that is, it may be supersaturated. Thus, after violent agitation, as in passing over a fall, oxygen may become mechanically entrained, and a sample taken immediately may show an amount several per cent above what normally would be expected. “Soda water,” which is water charged with carbonic acid under pres- sure, is an exaggerated case of supersaturation. On exposure to ordinary atmospheric conditions a supersaturated water gives off gas until equilibrium is established. Sometimes the quantity of gas given off is so minute that it can be measured only by the most delicate chemical methods; at others the action is vigorous enough to become manifest to the eye in the phenomenon of effervescence. Cases of 106 GEORGE C. WHIPPLE AND HORATIO N. PARKER water supersaturated with carbonic acid are very common. In out- door air the pressure of carbonic acid is so very low that the amount which is dissolved in water is very small. Ground water usually contains relatively large amounts of carbonic acid, and on exposure to the air yields it up until normal conditions of solution are estab- lished. This tendency to equalization makes it necessary to exer- cise great care in the determination of carbonic acid. Water may be freed of its dissolved gases to a considerable extent by boiling, but it is only by prolonged boiling in a vacuum that the last traces can be removed. In the early days of water analysis the dissolved gases were driven off in this manner and were collected and measured by the tedious process of gas analysis. The procedure still remains one of the most accurate, but on account of its inconvenience it has been superseded by simpler methods for the determination of dissolved oxygen and carbonic acid. WINKLER’S METHOD OF DETERMINING THE AMOUNT OF DISSOLVED OXYGEN IN WATER? Sample.—The sample from which the determination of dissolved oxygen is to be made must be collected with extreme care, as faulty, manipulation may easily cause an increase of oxygen by absorption from the air. A glass-stoppered bottle with narrow neck and hold- ing approximately 250 c.c. should be used. The exact capacity of the bottle with the stopper in place should be determined and, for convenient reference, scratched with a diamond upon the glass. The bottle can not be safely filled by pouring the sample into it, nor yet by sinking it in a pond and allowing the air to bubble out; it is necessary that the water enter at the bottom in a gentle current. If the sample is to be collected from a faucet the water must be made to enter the bottle through a glass or rubber tube which reaches to: the bottom of the bottle, the water being allowed to overflow for several minutes, after which the glass stopper is carefully pressed home so that no bubble of air is caught beneath it. If the sample is to be collected from a stream or pond the apparatus shown in Fig. I should be used. ” @ Berichte, KXI, 2843; 1888. See also Richards and Woodman, Air, Water, and Food, p 107; John Wiley and Sons, 1900, New York. A. H. Gill, Zech Quar., V, p. 250; Sia 3 Special” Rept. Mass. St. Bd. of Health on Purification of Sewage and Water, p. 722; 18 EFFECT OF GASES ON MICROSCOPIC ORGANISMS 107 E B Fig. 1. The sample bottle, 4, is provided with a rubber stopper through which pass two tubes, one reaching nearly to the bottom of the bottle and the other terminating at the bottom of the stopper. The longer one is attached to a rubber tube of length sufficient to reach the point where the sample is to be taken ; the shorter is attached to the tube of a large bottle, B, similarly provided with inlet and outlet. If suction is applied at the point D the water will enter at C, fill the bottle 4, and ultimately fill the bottle B. The bottle B should have-several times the capacity of A, and water should be drawn through A until a fair sample has been secured. If the sample is to be collected from a considerable depth a small lead pipe, which will serve as a weight and will neither collapse nor stretch, may be attached to C. It may be marked off in meters showing the depth of the sample. Suction may be secured with the mouth, but it is preferable to use an air pump. As soon as the bottle A has been filled with a satisfactory sample the rubber stopper should be removed and the glass stopper 108 GEORGE C. WHIPPLE AND HORATIO N. PARKER inserted as before. It is always advisable to ascertain the tempera- ture of the sample in order that the results of the determination may be expressed, if desired, in per cent of saturation. SOLUTION REQUIRED a. Manganous sulfate solution. Dissolve 48 grams of MnSO,. 4H.O in 100 c.c. of distilled water. b. Solution of sodium hydroxid and potassium iodid. Dissolve 360 grams of NaOH and 100 grains of KI in 1,000 c.c. = distilled water. c. Hydrochloric acid, sp. gr. 1.20. d. Sodium thiosulfate solution. Dissolve 24.827 Lee of chemic- ally pure recrystallized sodium thiosulfate (Na,S,O,.5H,O) in 1,000 c.c. distilled water. Dilute 100 c.c. of this solution to one liter with distilled water. This gives a N/too solution, each cubic centimeter of which is equivalent to .00008 grams of oxygen, or 0.055825 c.c. of oxygen at o° and 760 mm. pressure. Inasmuch as this solution is not perma- nent it should be standardized occasionally against a N/1oo0 solution of potassium bichromate as described in almost any work on volu- metric chemistry. e. Starch Solution —Mix a small amount of clean potato starch with cold water until it becomes a thin paste; stir this into 150 to 200 times its weight of boiling water and continue the boiling for a few minutes ; then let it stand and settle. It may be preserved by adding a few drops of chloroform. METHOD OF PROCEDURE Remove the stopper from the bottle which contains the sample and with a pipette long enough to reach the bottom of the bottle add approximately 2 c.c. of the manganous sulfate solution. Then, in a similar way, add approximately 2 c.c. of the sodium hydroxid potassium iodid solution. Replace the stopper in such a way as to leave no air bubbles, and mix the contents of the bottle. The man- ganous sulfate and sodium hydroxid react to form manganous hydroxid, a part of which at once combines with whatever dis- solved oxygen is present, to form manganic hydroxid. Allow the precipitate to settle; add about 2 c.c. of strong hydrochloric acid EFFECT OF GASES ON MICROSCOPIC ORGANISMS 10g and mix thoroughly. This reacts with the manganic hydroxid and liberates chlorin, which in turn liberates iodin from the potassium iodid. The amount of iodin liberated therefore depends upon the amount of dissolved oxygen present. Up to this point the process must be carried on at the time the sample is collected, but after the hydrochloric acid has been added and the stopper replaced, there is practically no further change, and the rest of the operation, which consists of the determination of the amount of iodin, may be conducted at leisure. Thus samples col- lected in the field are dosed with the three solutions and then taken to the laboratory where the examination is completed. The amount of liberated iodin is determined as follows: When the precipitate has nearly dissolved after the addition of the acid, the contents of the bottle are rinsed out into a flask and titrated with a N/too solution of sodium thiosulfate, using a few cubic centimeters of the starch solution as an indicator. It is best not to add the starch until the color has become a faint yellow. The titration is continued until the blue color has disappeared. CALCULATION AND EXPRESSION OF RESULTS There are three ways of expressing the amount of dissolved oxy- gen in water, namely: In parts per million by weight, in number of cubic centimeters of the gas per liter at 0° and 760 mm. pressure, and in per cent of saturation, that is, the per cent which the amount of gas present is of the amount capable of being dissolved by water at the same temperature and pressure. The following formulae may be used to ascertain the results ac- cording to the three methods: - -000087 1,000,000 807” Oxygen in parts per million= Vv v Oxygen in c.c. per Fe Bese . __-.0557882 X 1000 X100__ 5578.82 Oxygen in per cent of saturation= TT Ay WTC Va | UE N i i where 2=—=number of c.c. of aa thiosulfate solution. v==capacity of the bottle in cubic centimeters less the volume of the manganous sulfate and potassium iodid solution added (i. e., less 4 c.c.). 110 GEORGE C. WHIPPLE AND HORATIO N. PARKER O=the amount of oxygen in c.c. per liter in water saturated at the same temperature and pressure. See Table II. TABLE II AMOUNT OF DISSOLVED OXYGEN IN DISTILLED WATER WHEN SATURATED AT DIFFERENT TEMPERATURES FROM 0° To 30° c.3 (PREPARED BY ROSCOE AND LUNT. SEE JOUR. CHEM. SOC., P. 552, 1889. SEE ALSO SUTTON’S VOLUMETRIC ANALYSIS, P. 275) - OF C.C. 0. ° -c. ee PER Pp a LOGARITHM ee | ae ee LOGARITHM (Centigrade) ae | OF NUMBERS (Centigrade) psp OF NUMBERS 0.0 9.70 .986772 16.0 6.82 833784. 0.5 9.60 .982271 16.5 6.75 829304 1.0 9.49 .977266 17.0 6.68 824776: 1.5 9.38 -972203 17.5 6.61 .820201 2.0 9.28 .967548 18.0 6.54 -815578- 2.5 9.18 .962843 18.5 6.47 .810904 3.0 9.08 958086 19.0 6.40 .806180° 3.5 8.98 -953276 19.5 6.34 .802089: 4.0 8.87 .947924 20.0 6.28 797960 4.5 8.78 .943495 20.5 6.22 793790 5.0 8.68 .938520 21.0 6.16 789581 5.5 8.58 .933487 21.5 6.10 785330 6.0 8.49 .928908 22.0 6.04 781037 6.5 8.40 .924279 22.5 5.99 177427 7.0 8.31 .919601 23.0 5.94 773786 7.5 8.22 .914872 23.5 5.89 -T70115 8.0 8.13 910091 24.0 5.84 -766413 8.5 8.04 .905256 24.5 5.80 . 763428 9.0 7.95 .900367 25.0 5.76 760422 9.5 7.86 895423 25.5 5.72 -157396 10.0 7.77 .890421 26.0 5.68 754348 10.5 7.68 885361 26.5 5.64 751279 11.0 7.60 .880814 27.0 5.60 748188 11.5 7.52 .876218 27.5 5.57 745855 12.0 7.44 | .871573 28.0 5.54 743510 12.5 7.36 .866878 28.5 5.51 741152 13.0 7.28 862131 29.0 5.48 738781 13.5 7.20 .857332 29.5 5.45 136397 14.0 7.12 .852480 30.0 5.43 -734800 14.5 7.04 847973 30.5 5.40 732394 15.0 6.96 .842609 31.0 5.38 730782 15.5 6.89 .837588 For example, suppose the capacity of the bottle to be 264.5 c.c., the temperature of the water to be 10° C., and suppose that 28.6 c.c. 3 Below 5° the results are extrapolated and are not strictly accurate. The results given are calculated for aeration at an observed pressure of 760 mm. When the observed pressure is below 760 mm. 1/7g the value must be subtracted for every 10 mm. difference; and when the observed pressure is above 760 mm. a like amount must be added. EFFECT OF GASES ON MICROSCOPIC ORGANISMS Iil a thiosulfate were used in the titration. Then 80 X 28.6, _ Oxygen in parts per million= an: 8.78 or, : : PS Hy eco 2 Silk Oxygen in c.c. per liter= SE adie or, : : 5578.8 X 28.6 Oxygen in per cent of saturation—=—__—_——— = 78.7 per cent. ys Pp U 1 260.5 7-77 7 ih Pp The labor of calculation may be diminished by the use of loga- 55-788 v : ‘ 80. : rithms. Furthermore, inasmuch as in the first equation, - ec in the second, and 5578.8 in the third are constants for any particu- v lar bottle, it is convenient to have in a note-book, or upon the bottles themselves, the logarithms of these quantities. It is also desirable to bear in mind the following data: I c.c. of oxygen at o° and 760 mm. pressure weighs .001434 g. a : WGC thiosulfate solution—.o0008 g.==.055788 c.c. 0.7 ¢.c. oxygen per liter—1 part per million by weight (approx- imately ). I c.c. oxygen per liter==1.43 parts per million by weight (approx- imately). I cu. ft. oxygen=40.6 g. I gallon oxygen=5.52 g. SOURCES OF ERROR It is necessary to correct for the manganous sulfate and for the potassium iodid-sodium hydrate solution added, but if the precipi- tate is allowed to settle before the acid is added it is not necessary to correct for the latter, as the water displaced has already given up its oxygen. In practice perhaps the greatest source of error is carelessness in collecting the sample, thus allowing atmospheric air to become en- trained. This is particularly the case when the amount of oxygen is small. Supersaturation with oxygen, however, seldom occurs. To some extent the amount of organic matter interferes with the Ii2 GEORGE C. WHIPPLE AND HORATIO N. PARKER result by exerting a reducing action, and high nitrates, also, ma- terially affect the result. These sources of error are discussed in Winkler’s paper; in actual practice it is seldom necessary to take them into account. Neither is it necessary to correct for atmos- pheric pressure when expressing the results in number of cubic centimeters per liter. METHOD OF DETERMINING THE AMOUNT OF CARBONIC ACID IN WATER Although water has a large capacity for absorbing carbonic acid, as stated above, yet the pressure of the carbonic acid in the air is so small that under ordinary atmospheric conditions the amount dis- solved is very slight. When water which contains carbonic acid comes in contact with the carbonates of calcium and magnesium it is supposed to unite with them to form the bicarbonates of these metals,—salts which are readily soluble. The normal carbonates, themselves, but are slightly soluble. If the amounts of the normal carbonates are in excess, all of the dissolved carbonic acid will be used up, but if the carbonic acid is in excess the quantity of it remaining in solution will be proportional to the pressure of the carbonic acid in the atmosphere. Inasmuch as most surface and ground waters come in contact with the carbonates of the alkaline earths during some part of their history, it follows that natural waters ordinarily contain carbonic acid in three different conditions. That part which simply remains in solution uncom- bined with any base is termed the “free carbonic acid”; if it is combined directly with the bases to form the normal carbonates (CaCO, and MgCO,) it is called the “fixed, or full-bound car- bonic acid”; while if it is combined indirectly with the normal carbonates to form the bicarbonates it is known as the “‘half-bound carbonic acid.” The sum of the three gives the total carbonic acid. When water which contains carbonic acid in these three forms is heated the free and half-bound carbonic acid is given off, and is sometimes referred to as the volatile carbonic acid. In the analysis of water for sanitary or industrial purposes it is sometimes necessary to ascertain the amount of carbonic acid in each of the three forms, but in the study of biological problems it is chiefly the free carbonic acid which is involved. Various methods have been suggested for the determination of the carbonic acid. These have been recently discussed in an excellent paper by Ellms and Beneker EFFECT OF GASES ON MICROSCOPIC ORGANISMS Lr3 in the Journal of the American Chemical Society, Vol. XXIII, No. 6 (June, 1901), to which the reader is referred. The free carbonic acid may be easily determined by titration with a solution of sodium carbonate, using phenolphthalein as an indi- cator. As soon as the sodium carbonate has combined with all the free carbonic acid present, with the formation of bicarbonate of soda, any further addition will produce a pink color. In case there is no free carbonic acid present the phenolphthalein will produce a pink color when the first drop of sodium carbonate is added, or even be- fore it is added at all. The details of the method as practiced by the authors are as follows: Sample.—lf the determination can not be made at the time the sample is collected, the bottle should be filled completely, without leaving any air space under the stopper. It is preferable to collect the sample in the receptacle in which the titration is to be made. The required solutions are prepared as follows: Sodium Carbonate Solution (Na,CO, a) A normal solution of sodium carbonate is first made by dissolving 53 g. of the freshly fused, chemically pure salt in one liter of freshly boiled distilled water. This should be kept in a hard glass bottle N : : Se OE with glass stopper. The a solution used in the titration is made from this by dilution with boiled distilled water. As this solution easily absorbs carbonic acid from the air and thereby changes to bicarbonate, it is important that it be exposed to the air as little as possible. It is even advisable to supply the burette by side connec- tion with the supply-bottle and to provide tubes filled with caustic lime upon both the supply bottle and burette, in order that the car- bonic acid may be removed from the air before it comes in contact with the solution. The burette should be graduated to 0.1 c.c. Phenolphthalein Indicator. This is prepared by dissolving 0.5 g. of the powdered salt in 100 c.c. of 50 per cent alcohol. A few drops of a weak solution of so- dium hydrate should be cautiously added until a faint pink color is found, which just disappears after shaking. For use this may be kept in a small glass-stoppered bottle with a pipette stopper. Mode of Procedure.—The determination is made by putting 100 c.c. of the sample in a 100 c.c. Nessler tube with 10 drops of the phenolphthalein indicator. The sodium carbonate solution is then II4 GEORGE C. WHIPPLE AND HORATIO N. PARKER added from a burette and stirred until the faint pink color which appears remains permanent. The difference between the burette readings before and after titration expressed in tenths of a cubic cen- timeter gives the amount of carbonic acid in parts per million. For example, if the burette readings are 31.3 and 32.0, the difference is 1.6, and the corresponding amount of carbonic acid is 16 parts per million. If due precautions are taken the results should be correct to about one part in a million. The greatest sources of error in the method come from the loss of carbonic acid to the air and from the absorption of carbonic acid by the solution of sodium carbonate. The latter may be prevented by adopting the precautions described above, while the former may be reduced to a minimum by making the titration as soon as pos- sible after collection, and by collecting the sample in the tube in which the titration is to be made. Great care should be used in stir- ring the solution. In fact, instead of stirring, it is often better to mix the added solution by rapidly whirling the tube in such a way that it describes the surface of a cone. If a stirring rod is used it should be bent so that at the lower end there is a circle with its plane at right angles to the rod. Exposure to the air ordinarily results in loss of free carbonic acid, which may be illustrated by the following experiment: A series of samples was collected and carbonic acid determinations made in the field, while a corresponding series was collected and sent to the laboratory, where determinations were made at the end of twenty-four hours. A third series was allowed to stand the same length of time, but without being stoppered. The results were as follows: AMOUNT OF CARBONIC ACID IN PARTS PER MILLION Determination made ate Determination made] after standing 24 pps eager at the time of col-| hours with tu Ouro Rceh aie lection completely filled) oy noced t the ‘a and stoppered P Oe ene Surface water No. 1..... 8.8 9.0 1.5 Surface water No. 2..... ee 1.5 0.5 Surface water No. 3..... 0.5 0.5 0.5 Ground water No. 1..... 7.0 6.5 1.5 Ground water No 2..... 7.0 7.0 1.0 Ground water No. 3..... 7.0 7.0 1.0 Ground water No. 4..... 2.5 2.0 —1.0 EFFECT OF GASES ON MICROSCOPIC ORGANISMS I1l5 ~ At another time carbonic acid gas was passed through distilled water until the water contained 45.5 parts per million. Portions of 200 c.c. were then put in completely filled, tightly stoppered bottles, and in open bottles exposed to the air at different temperatures. The results were as follows: AMOUNT OF CARBONIC ACID (PARTS PER MILLION) At beginning|After standing} After standing of experiment 3 hours 20 hours Bottle filled and tightly stoppered..... 45.5 44.0 44.5 Bottle unstoppered, temperature 17.0° C. 45.5 26.0 0.5 Bottle unstoppered, temperature 20.0° C. 45.5 23.5 0.5 Bottle unstoppered, temperature 37.0° C. 45.5 15.0 0.3 Thus in less than twenty-four hours practically all of the carbonic acid had been given off to the air. It is not even necessary that the bottle shall be unstoppered in order that a loss of carbonic acid may occur. If the bottle is only partially filled, there oftentimes will be a loss to the air in the bottle. Thus a water which contained 15 parts of carbonic acid per million was distributed into half-filled 8-oz. bottles. The first bottle was tested at the end of twenty-four hours; the second at the end of forty-eight hours, and so on, with the following results: AMOUNT OF CARBONIC ACID BEEN ENE RTUR ENTIRE erate op hc -9-. on; wtoyo v5 eel suche reheeey raatarelata ome males oe 15.0 PONG OL AT HOULS Hie ccs cls sala ars eke Ree eae ais siaeahs 13.5 ENE Gr lyon 8319010) so A SIE tos CIC ois Dom Orme DOR or One 8.0 Atretidhotel nOUnrSee < 5.5.4 sills Sasctatolatabe an deelaisl oiekeleisie Sua laiersicl tie 2.5 AGFeTI COM GO OUMES HS eis ee Uo ee Rie oiain eee niateleraratv aes 1.8 The danger of loss of carbonic acid while pouring the sample back and forth through the air is illustrated by the following experi- ment: Carbonic acid was passed through water until 63 parts per million were dissolved. A portion of the water was then poured back and forth four times and tested. It was found to contain 57.5 parts, showing a loss of 9 per cent. The amount of carbonic acid given up in this way depends in part upon the pressure of the carbonic acid in the air. On exposure to the air a sample of boiled distilled water will absorb from it a certain amount of carbonic acid. Out of doors this is seldom greater than 116 GEORGE C. WHIPPLE AND HORATIO N. PARKER 0.5 parts per million, but in the air of a laboratory, when gas is used freely, it sometimes amounts to I.5 or even 3.0 parts. Ellms and Beneker mention an instance where it was I5 parts per million, but in this case a carbonic acid generator was in use in the laboratory. There is, in short, a tendency for the carbonic acid in the air and in the water exposed to the air to come to an equilibrium. This must occur in natural bodies of water as well as in small samples, and it doubtless plays an important part in the economy of nature. It sometimes happens that there is no free carbonic acid present, in which case the water is neutral to phenolphthalein ; or the water may be alkaline to phenolphthalein, showing that some of the car- bonates are in solution as normal carbonates instead of as bicarbonates. If it is desired to obtain the amount of carbonic acid in the three A Oh tals 40 ae states mentioned, a second titration should be made with se HS@y using methyl orange (cold), or preferably lacmoid (hot) as the indicator. It is best to express the results of all chemical analyses of water in parts per million, that is, in milligrams per liter, but in case it is desired to express the amount of carbonic acid in cubic centimeters per liter, the following table* will be found convenient: TABLE III WEIGHT IN MILLIGRAMS OF A CUBIC CENTIMETER OF CARBONIC DIOXID FROM 746 to 778 MILLIMETERS PRESSURE AND FROM 10° To 25° Cc. CORRECTED FOR THE TENSION OF AQUEOUS VAPOR. + Py MILLIMETERS a HS | l | x “| 946 | 748 | 750 | 752] 754] 756] 75S | 760} 762 | 764] 766) 768 | 770| 772 | 774) 776} 778 410°} 1.839] 1.844] 1.849]1.854/1.859/1.864)/1.869)1.874/1.879}1.884/1.889]1.894/1.899/1.904|1.909)1.914)1.919 11 | 1.831) 1.836) 1.841/1.846)1.851/1.856/1.861/1.866) 1.871)1.876)1.881/1.886/1.891/1.896/1.901/1.906/1.911 12 | 1.823] 1.828] 1.833/1.838/1.843]1.848)1.853/1.858)1.863/1.868/1.873/1.873}1.883/ 1.888) 1.893) 1.898) 1.903 13 | 1.815] 1.820] 1.825/1.830/1.835|1.840/1.845/1.850/1.855/1.860) 1.865|1.870/1.875/1.880)/1.885)1.889|1.894 414 | 1.807) 1.812) 1.817/1.£22]1.827|1.832]1.837|1.842|1.847/1.852/1.856|1.861/1.866/1.871| 1.876) 1.888) 1.886 45 | 1.799] 1.804) 1.809]1.814/1.818]1.823/1.828/1.833|1.838|1.843/1.848)1.853]1.858 1.863 1.868} 1.872/1.877 16 | 1.791} 1.795] 1.800]1.805]1.810) 1.815}1.821|1.825)1.830/1.835/1.839|1.844/1.849/1.854 1.859}1.864) 1.869 17 | 1.782] 1.787) 1.792/1.797|1.801|1.806}1.811/1.816]1.821/1.826|1.831|1.836/1.841/1.846)1.851)1.855/1.860 18 | 1.774] 1.779] 1.784/1.788|1.793|1.798/1.803|1.808/1.813/1.818|1.823|1.828/1.832]1.837|1.842/1.847/1.852 19 | 1.785} 1.770] 1.775/1.780]1.785|1.790|1.794/1.799|1.804/1.809/1.814/1.819|1.823/1.828/1.833|1.838/1.843 20 | 1.757| 1.761] 1.766]1.771|1.776|1.781)1.786| 1.791) 1.795/1.800|1.805)1.810) 1.815/1.820) 1.825)1.829)1.834 21 | 1.748] 1.753) 1.758/1.623)1.767|1.772/1.777|1.782) 1.787}1.792/1.797|1.802|1.806/1.811/1.816)1.820)1.825 22 | 1.739} 1.744] 1.749]1.754/1.759|1.764/1.769] 1.773) 1.778]1.783]1.787|1.792|1.797|1.802/1.807|1.811/1.816 23 | 1.731) 1.735] 1.740|1.745}1.750}1.755/1.760]1.764|1.769|1.774/1.778)1.783 1.788 1.793} 1.798|1.802/1.807 24 | 1.721) 1.726] 1.731/1.736]1.741)1.746)1.751/1.755|1.760/1.765|1. 769} 1.774) 1.779}1.184)1.789}1.793)1.798 25 | 1.713) 1.718] 1.722/1.727|1.732|1.737|1.741|1.746|1.751|1.756/1.760| 1.765) 1.770)1.775|1.780)1.784/1.789 4Ellen H. Richards and Alpheus G. Woodman, Air, Water, and Food, p. 196. John Wiley & Sons, New York, 1900. EFFECT OF GASES ON MICROSCOPIC ORGANISMS ELT AMOUNT OF OXYGEN IN NATURAL WATERS UNDER VARIOUS CONDITIONS The oxygen dissolved in water is derived primarily from the at- mosphere. If distilled water, freed of oxygen by boiling, stands exposed to the air it absorbs oxygen and in a few hours becomes sat- urated, or, to use the popular expression, aerated. This is accom- plished even more quickly if the water falls through the air in drops. Roscoe and Lunt obtained the standard aerated water used in the preparation of Table II as follows: Two glass-stoppered quart bot- tles were half-filled with distilled water and shaken for five min- utes, the air being renewed several times by filling one bottle with the contents of the other and dividing it again into two portions. Finally one bottle being filled, the temperature was taken and also the barometric pressure, after which the bottle was allowed to stand stoppered for half an hour to get rid of minute air bubbles. Roscoe and Lunt’s figures are probably somewhat more accurate than those given by Bunsen and Winkler. In the case of natural waters exposed to sunlight there is another source of oxygen, namely aquatic vegetation. It is a well-known fact that chlorophy!laceous plants exposed to sunlight exhale oxygen. This is true of aquatic as well as terrestrial plants, and of the micro- scopic organisms as well as the larger forms. Several years ago T. Chalkley Palmer® described an interesting method for demonstrating this fact in the case of the diatom Eunotia major, Rabenhorst. It was based upon the color changes produced in hematoxylin by car- bonic acid and oxygen. When this indicator is dissolved in water which contains oxygen it has a rosy pink color, but if one breathes into the solution through a glass tube the color will change to a light brown. If now the solution be poured back and forth from one beaker to another the pink color will return, because the carbonic acid will have been liberated and oxygen acquired. Palmer placed his growing diatoms in a test tube provided with a capillary tube which passed through the stopper. The test tube was held inverted with the capillary point under the surface of a jar of water. The water was first colored pink by the addition of a watery solution of hematoxylin. The color was then changed to a light brown by 5 Proc. Acad. of Nui. Sci. of Philadelphia, p. 142. Feb., 1897. 8 118 GEORGE C. WHIPPLE AND HORATIO N. PARKER charging the water with carbonic acid from the lungs. The appa-— ratus was then exposed to sunlight, and in the course of fifteen minutes or half an hour the color of the liquid in the test tube had become pink because of the liberation of oxygen by the growing diatoms. Later the color became a brilliant red. M. Albert-Levy, Directeur du Service Chimique, l’Observatoire de Montsouris, Paris, has also observed that if samples of water containing algae are kept exposed to sunlight the amount of dis- solved oxygen will be increased, and he cites an example where water from the Vanne, which at the time of collection contained 11.1 parts per million, after nine days was found to contain 20.2 parts per million, and after sixty days 39.7 parts. If these figures are correct we have here an example of supersaturation. While plant life and the atmosphere are supplying oxygen to sur- face waters, other forces are at work consuming it. All forms of aquatic animal life, both large and small, require oxygen, and most bacteria are likewise dependent upon it. Some oxygen besides is used up without the direct intervention of vital processes in the direct oxidation of organic matter. The fact that minute animal organ- isms, stich as snails, crustacea, rotifera, etc., use up oxygen may be illustrated by placing them in one of Palmer’s tubes and noting the change in color of the hematoxylin solution from red to brown. The exhaustion of oxygen by a culture of bacteria in a hermetically sealed jar may be also shown by the use of this indicator. The fact that de- composition of organic matter is accompanied by loss of dissolved oxygen has given rise to the so-called “incubation test” for determin- ing the quality of water. The sample, which must completely fill the bottle, is kept for several days in the dark at a temperature favor- able to rapid bacterial development, and the amount of dissolved oxygen determined before and after the incubation. The greater the amount of organic matter present and the larger the number of bacteria in the original water, the more rapid will be the exhaustion of the oxygen. M. Albert-Levy found that a sample of water from the river Seine which, when collected, contained 10.6 parts per mil- lion of dissolved oxygen, lost all of its oxygen after standing fifteen days. While this test is too crude and too inconvenient to be of practical use, it is interesting as an illustration of the point in ques- tion. In general, therefore, it may be said that decomposition of organic matter in water means loss or even exhaustion of dissolved oxygen. EFFECT OF GASES ON MICROSCOPIC ORGANISMS 11g Surface waters which contain but little organic matter and which are exposed to the air are usually saturated or almost saturated with oxygen. This is equally true of running brooks, large streams, small pools, and the surface strata of large lakes. Such waters sel- dom contain less than 80 per cent of the amount required to saturate them. Even if saturated, however, the actual amount of oxygen present varies considerably at different seasons on account of the effect of temperature upon the solubility. This is illustrated in one of the diagrams on Plate XX, which shows the seasonal changes in the temperature of surface waters at Boston, Mass., and the correspond- ing changes in the amount of oxygen present in a fully saturated water. (See Table IV.) Gill® has shown that at times natural waters may be supersaturated with oxygen, and the same fact has been ob- served elsewhere. Horton’ has stated that in certain streams the water is normally supersaturated during the months of July and August, and he attributes this to the influence of growing plants. TABLE IV TABLE SHOWING THE AMOUNT OF OXYGEN IN WATER WHEN SATURATED AT DIFFERENT SEASONS OF THE YEAR AT BOSTON, MASS. AMOUNT OF OXYGEN MONTHS AVERAGE TEM- = f bi = PERATURE, oO. ° cubic Parts r CENTIGRADE centinctets Pallien Jannary,..-6- S35 8.92 12.8 February 3.4 9.01 12.9 Marchi. 2... ie) 8.90 UAT Aprile ro ic 2 7.4 8.24 11.8 Mayo seyeieton 13.6 - 7.19 10.3 June3s2 55: koa 6.39 9.1 Jilliygey sete scess 22.0 6.04 8.6 AUISTISE ;.. i: 22.1 6.03 8.5 September ... 20.0 6.28 9.0 Octobera.css- 14.5 7.04 10.0 November.... 10.4 7.69 11.0 December.... 6.7 8.38 12.0 If water contains much organic matter, bacterial decomposition may take place, especially during warm weather. This results in the 6 (1. c.) 7Second report of an investigation of the rivers of Ohio by the State Board of Health 1899. 120 GEORGE C. WHIPPLE AND HORATIO N. PARKER reduction of the oxygen or even in its entire disappearance. Shallow ponds with muddy bottoms or stagnant pools and sluggish streams with an excess of aquatic vegetation often lose their oxygen during the summer. Sewage, which is very rich in organic matter, loses its oxygen in a very few hours, after which putrefaction sets in. When sewage is allowed to enter a stream the dissolved oxygen is reduced by an amount corresponding to the strength of the sewage and the ratio of its volume to that of the stream. The Merrimac River receives the sewage of the city of Lowell, besides that of other cities, and although the volume of the stream is large, the diminu- tion in the amount of dissolved oxygen in the water at Lawrence, nine miles down stream, is marked. Table V gives by months the per cent of saturation of the water at Lawrence for the years 1893-95. It will be observed that during the summer months there was a material reduction in the amount of oxygen. One of the dia- grams on Plate XX shows the effect of the sewage of Paris upon the water of the river Seine. At Choisy-le-Roi, above the entrance of the large sewers, the water is almost saturated for the greater part of the year, and even in summer does not fall far short of saturation; but at Argenteuil, below the city, the amount of oxy- gen is at all times below that of saturation, and during the sum- mer it is almost exhausted. The same fact is illustrated by Table VI, which shows the progressive diminution of the amount of oxygen between the two stations named, and in parallel series the increas- ing foulness of the river as illustrated by the chlorin, the organic matter, and the disappearance of oxygen during the forty-eight hours’ incubation at 33° C. TABLE V TABLE SHOWING THE AMOUNT OF DISSOLVED OXYGEN IN THE WATER OF THE MERRIMAC RIVER AT LAWRENCE, EXPRESSED IN PER CENT OF SATURATION JAN. | FEB. | MAR.| APR. | MAY | JUNE/| JULY} AUG. |SEPT.} OCT. | NOV. | DEC. BOS. cweseienss 91 | 96| 91 | 100} 86} 75| 52} 69| SL} 86] 95] 98 LEGS eet ae seeded el lease | esi) MGMT TG) Tet SGN ST SS a ieee en BOD Anhecieiee cists 83 | 82| 79; 98] 88| 62} 59] 55 | 50] 68] 99 EFFECT OF GASES ON MICROSCOPIC ORGANISMS I21 TABLE VI TABLE SHOWING THE AMOUNT OF DISSOLVED OXYGEN AT VARIOUS STATIONS ON THE RIVER SEINE ABOVE AND BELOW THE CITY OF PARIS. AVERAGE OF TEN YEARS’ OBSERVATION BY M. ALBERT-LEVY DISSOLVED OXYGEN IN PARTS PER MILLION ORGANIC " ee) SEA Ci Pe | eee D a ta | Attime of | After48 | Percent | MATTER acne | collection} hours at used up | ofsample| 33°C. during in- cubation 2.2 a Se Gharsy-le-Rot ..:........:.. 10.4 7.8 25 Sil 6.0 WISHe PTV TY:s cases sice ne AR 10.8 8.8 18 2.8 7.0 Usine d’Austerlitz.......... Lo A0E7 8.4 21 2.9 7.0 sme Chaillot... 5./% 3:22: 10.0 75 25 ae TA Usine de Suresnes.........., 8.6 5.3 38 33.11 8.0 Usine d’Argentenil......... ea 2.4 63 4.5 11.0 The amount of dissolved oxygen in large bodies of water varies with the depth. Many observations illustrating this fact are given in Table VII. Near the surface and as far down as the water is kept in circulation by the wind the per cent of saturation is usually above 80 and is often 100, while in the stagnant portion of the water below the thermocline the oxygen diminishes toward the bot- tom, sometimes gradually and sometimes suddenly. This ex- haustion of oxygen has been fully discussed in the reports of the Massachusetts State Board of Health, the Boston Water-works, and elsewhere.’ During the winter, as in the summer, there is an ex- haustion of oxygen at the bottom of stagnant lakes and often a partial exhaustion at the surface immediately under the ice. This is illustrated by figures given in Table VII. The extent to which the oxygen is used up at the bottom of deep lakes depends chiefly upon the condition of the soil at the bottom. If it is clean sand or gravel the water may remain at least partially aerated for a long time even though the lake is thermally stratified, but if there is a deposit of organic matter decomposition sets in and robs the water of its oxygen. 8See Microscopy of Drinking Water by G. C. Whipple. John Wiley & Sons, New York. 122 GEORGE C. WHIPPLE AND HORATIO N. PARKER TABLE VII fABLE SHOWING THE AMOUNT OF DISSOLVED OXYGEN AT VARIOUS DEPTHS IN CERTAIN PONDS AND RESERVOIRS IN MASSACHUSETTS. COMPILED FROM THE REPORTS OF THE MASSACHUSETTS STATE BOARD OF HEALTH, THE AN- NUAL REPORTS OF THE BOSTON WATER-WORKS, ETC, DISSOLVED POND DATE ISLES UR aoe Pives eet BEET TIGRADE |Per cent of saturation Jamaica Pond): o%)2).)6o<.ceeee se June 11, 1891 0 25 100 10 20 100 20 20 80 30 12 59 40 12 al BO | coe 0 9 MET seed cose 0 June 25, 1891 0 21.7 100 35 5.8 37 40 5.8 0 50 5.6 0 July 14, 1891 0 24.0 100 10 23.8 100 20 12:3 49 30 5.8 29 35 5.6 4 40 5.4 0 47 5.2 0 Jan. 24, 1893 0 0 98 (surface 10 2 i frozen ) 20 2.1 89 30 2.2 72 40 2.2 19 44 2.2 0 Wake: Cochitnates 2.62). seis) Aug. 17, 1891 0 23.6 79 10 19e0 84 20 12.1 36 30 9.6 21 40 Soil 21 45 weil 2 50 7.6 0 57 (ent 0 Feb. 14, 1893 0 0 100 (surface 10 3.8 100 frozen) 20 2.8 100 30 2.8 94 40 2.8 83 45 2.8 60 50 2.9 49 55 2.9 37 Sept. 18, 1890 0 21.0 97 15 21.0 88 : 10.0 20 40 9.4 12 50 Kec 0 60 7.5 0 EFFECT OF GASES ON MICROSCOPIC ORGANISMS 123 TABLE Vil—Continued | DISSOLVED TEMPERA-| OXYGEN POND DATE ee TURE CEN- TIGRADE |Per cent of saturation ake Cochitnate............. Sept. 28, 1891 0 21.0 90 10 14.0 81 20 14.5 33 30 11.0 9 40 11.0 8 50 10.0 0 56 9.0 0 Mestic: Hake es .clelos sale March 8, 1893 0 0 60 (surface 10 1.2 64 frozen ) 20 2.1 68 30 2.2 58 40 2.2 55 | 50 2.2 50 54 2.4 49 | 60 2.4 43 64 2.4 25 66 pana | 13 68 2.7 0 74 2.7 0 Reservoir No. 4, Boston...... Aug. 20, 1891 0 23.6 85 10 21.6 84 20 16.6 28 | eae nce 27 35 12.6 16 361g 12.6 15 Feb. 14, 1893 0 i) 100 (surface 10 3.4 100 frozen ) 20 3.8 92 25 3.9 78 30 3.7 60 | Sept. 25, 1890 0 18.2 92 10 18.0 87 20 15.9 18 29 12.3 7 Oct. 1, 1891 0 22.0 88 10 21.5 84 15 21.0 83 20 15.0 9 25 14.5 7 31 12.0 4 Reservoir No. 3, Boston...... Aug. 20, 1891 0 23.0 86 6 23.6 85 12 21.6 59 fC Sea a at cea 0 BY | Pale ss as uae 0 bY Raed ae nets 0 bE BM It Nea ele ego 0 21 dit (eal! 0 Jan. 30, 1893 0 2.2 81 (surface i) 2.1 62 frozen) 10 2h 45 124 GEORGE C. WHIPPLE AND HORATIO N. PARKER TABLE VIIl—Concluded DISSOLVED TE -| OXYGEN POND DATE specie ea SS =o TIGRADE |Per cent of saturation Reservoir No. 3, Boston...... | Jan. 30, 1895 14 2.3 44 Reservoir No. 2, Boston...... Jan. 30, 1833 0 1.2 86 (surface 5 13 79 | frozen) 10 ass 32 Scott Res., Fitchburg........ | July 29, 1891 0 21.4 87 10 18.8 79 20 16.2 32 25 10.2 11 30) ieee 0 CS eso Fic: 0 Glenn Lewis Pond, Lynn ....| June 26, 1891 0 24.2 100 Di eterna 100 Cee 100 10) Vos 0 1344; 17.2 0 Jan. 26, 1893 0 2.8 7 (Surface frozen) 5 3.3 6 Walden Pond, Lynn......... Jan. 26, 1893 0 3.8 64 (surface 5 3.8 26 frozen ) 10 oD 20 | 15 2.8 24 Brocton Reservoir........... Mar. 15, 1893 0 0 90 (surface 5 3.0 80 frozen) 10 4.0 60 12 aod 28 14 4353) 20 16 338) 4 18 338) 0 Van Horn Res., Springfield..| July 16, 1891 0 24.1 100 7 24.1 100 14 24.1 46 1 WAP Fehr si 0 20. (4-26 eee 0 2S) ees 0 EFFECT OF GASES ON MICROSCOPIC ORGANISMS 125 TABLE VIII TABLE SHOWING THE AMOUNT OF DISSOLVED OXYGEN IN CERTAIN OHIO RIVER WATERS Scroto RIVER TEMPERATURE| DISSOLVED STATION DATE CENTIGRADE | 0 fo SATUR- Kenton above: towtl 2). ..\.ioscsse sae bbs Carag Aan Bae OS ee Mee aL AA eater July 16.. 25.5 91 Aug. 20.. 21 85 Sept. 25.. 16 84 Oct7) 234. 15 114 Decw)) He: i $4 Renton. below: tOwils ssc. cisiia sees cee {Peary Dice ara een Seti le lieve el hea ee July 16.. 25 114 Aug. 25... 23.5 169 Sept. 25.. 18.5 72 OEE oe: 16.5 159 Decl ts: 3.79 98 Girls’ Industrial Home, above......... Ocenisis:. Lies 106 Oct. -26.. 15.5 99 Dec. | 2:. 2.5 94 Girls’ Industrial Home, below......... Ocho ies 17 113 Oct! + 262). 17 116 OCH 265). 17 118 Dee wien. SN ad Ee UNM Da ed Dee) j26:): 3 91-83 WV ANOLE GrOVEs ccs. 1065.0 2 leis oe eceich & TIS LEE Ve cparsratete a etere | icrelcsopeenae mre Janes) ii: 27 108 Aug. 23.. 23 88 Sept. 23.. 17 89 Octy 2). 16 79 Dec.) 73. 2.5 85 HOSES OLIGO SISOS REE DER Seem PaReIIE ar Jue: sel stood oie lair sens eee July 17.. 25.5 93 Aug. 23.. 23 101 Sept. 23. 19 86 @etey nie 16 73 Deen ols: 2.5 86 Columbus, Sandusky St. Bridge....... Jarre HS SAE srabelleens ane Hs ON settee jpotls7) Wl 27.5 | 118 Aug. 23.. 22.5 42 Sept. 23.. 20 91 Oct.) (Le. 16 96 Dee Kiss 3 89 Coiumbus, Frank Rd. Bridge......... Gal) Pia : 23.5 9 (below sewers) Aug. 23... 23 21 Sept. 23. 19.5 0 Oct, 2. 17 0 : 1D frente Cane a) 78 SHadeville Bridge... 0.006. .os cls Vee sige: Aji ee ieee 23 46 (apparently below town) Aug. 23... 22.5 100 Sept. 23.. 19 58 Oct. 22.. 17 0 Dec yaa 4.5 81 126 GEORGE C. WHIPPLE AND HORATIO N. PARKER TABLE VIll—Continued Scioto RIVER—Continued TEMPE DISSOLVED STATION DATE CEN Gee °/o SATUR- Cuchville, Main St. Bridge............ July 22... 26 57 Aug. 26... 22.5 57 Sept, aioe. 18 71 Oeil 2oen 14 32 ID Io ie Ware 6 59 LITTLE Scioto RIVER Darion, ‘above tOwi a ce esis to oii: July 15... 26 116 Aug. 24 23 114 Sept. 24 20 50: Oct. 23 a 50 Dees, Ne 1 82 Marion, below, tows \ssics ose esis ellyar lan 26 1125 Aug. 24.. 23 46 Sept. 24.. 21 61 Oct. 23.; 1i.2 72 DEC ler 2 27 CLETANGY RIVER Galion above tOwi)s)cie).<)- Aeleries ote sleunle itliyan lo ever 22.5 147 Aug. 24... 22.5 123 Sept. 24... 21 107 Oct, 22:0" 12 74 Nov. 30... 3 90 Galion, below town.......:-.....52..- July eto 24 51 Ang... 4. )5). 20 36 Sept. 24.. 18 77 Oets gzehe 13 0 Noy. 30... 4 72 Delaware) above town: <<: j/s)6..)2 << July, lore. 22 87 Aug. 24... 21 76 Sept. 24.. 15 84 Oct?* '2e<. 14 87 Deets, 2.4% 1 82 Delaware, below town ....:.........-. Tietbvae URS Se 24 91 Aug. 24... 19 cl Sept. 24.. 13 79 Oct. 7:26... 14 99 Dees 7203 2.5 89 Cletanoy (Park, ABOVE. cs basin ces os ni Yfrtyye ay Sr 24.5 85 ; Aug. 24 22.5 80 Sept. 23 17 74 Oct ak 15.5 61 Dee’! 3.0% 3 83 Columbus, Dublin Bridge............. july» Az... 27 109 | Aug. 23 23 97 Sept. 23 21 129 Oct 3 16.2 93 Dec, 3 3 89 EFFECT OF GASES ON MICROSCOPIC ORGANISMS 127 TABLE VIlI—Concluded MAHONING RIVER ye TEMPERAT DISSOLVED STATION DATE CENTICR. y hey 0 seers | Alliance, above Pat. St. Bridge........ rly 2ae: 23 66 Sept. 2.. 23 94 Sept. 29.. 19 97 Oct 273" 14 80 Nov. 26... 11 81 PANiitar ce. bel OWiss/:-5 4's iciicnew Sia clerisers Sele July <24.... 25 15 Sept. 2... 25 81 Sept. 29.. 19 70 Oct: 27. 16 35 Nov. 26.. 8 90 WWiatren’, (ADOVE wha). ctecriccieise ce micielsia teats July 28.. 23 68 Aug. 28.. 23.5 99 Sept. 30.. 16 90 Oct. 5 23. 13 82 Nov. 27.. 7 83 Wranrenibelowyacacckere« oceiss ee cee July2oee 23 76 Aug. 28.. 23 66 Sept. 30.. 16 72 Og 28°: 14 54 Nov. 27.. 8 90 MUMMIES eo te cil ee cious oble's Slo td's ove pets July 24.. 23 70 Aug. 28.. 21.5 92 Sept. 30.. 15 89 Oct. 28... 13 70 Nov. 27... 8 81 Wonttestown, above: ... 2... 2202566: July 23... 21.5 86 Aug. 28.. 26 82 Sept. 30.. 20.5 93 Oet. ) 28x. 16 76 Nov. 217... 7.5 90 Ree LOM BORG irc clsais cists oe .c's s nc/sle July 238... 21.5 72 Vvitess Weegee 26 65 Sept. 30.. 22 77 Oct. 28... 18 51 Nov. 27.. 7 84 The amount of dissolved oxygen in certain river waters of Ohio as given above in Table VIII, is taken from the Thirteenth Annual Report of the State Board of Health. The tap water in most towns and cities where surface supplies are used is usually well aerated, as shown in Table IX, but in the case of water rich in organic matter or in micro-organisms a loss of oxy- gen sometimes occurs in the pipes of the distribution system, as Table X illustrates. 128 GEORGE C. WHIPPLE AND HORATIO N. PARKER TABLE IX TABLE SHOWING THE AMOUNT OF DISSOLVED OXYGEN IN THE PUBLIC WATER-SUPPLIES OF CERTAIN MASSACHUSETTS CITIES AND TOWNS WHERE SURFACE WATER IS USED. COMPILED FROM REPORT FROM MASS. ST. BD. OF HEALTH FOR 1898, P. 367 ms DISSOLVED OXYGEN CHEY/OR LOWN (Per cent of saturation) BOStOM ene Haeiete o elena creme 84 Stouciton Vu esc sa ae. 73 New sbedtord aencncsiemes eos 54 INGEWOOG eis coerce cians oe 100 Paltrent tase ie aoa eimeeee: 92 CHIcoHee it L se once gees 97 Wa wrence en aia cee ee 85 Concord yesh cee eae | 75 Mall (Ravers seston 98 Rockland sis eee ha Se 83 Syol gia ten eto) (6 MAAS Be reas AR onde 97 | TABLE X TABLE SHOWING THE DECREASE IN THE AMOUNT OF OXYGEN IN THE PIPES OF A WATER-WORKS DISTRIBUTION SYSTEM. COMPILED FROM MASS. ST. BD. OF HEALTH REPT. 1891, P. 379 | DISSOLVED OXYGEN TEMP. |per cent of saturation Large Ludlow res., Springfield, Mass-|Surface |July 16, 1891) 24.1°C. 98 Large Ludlowres., Springfield, Mass-|Bottom.|July 16, 1891] 23.6 61 Small Ludlow res., Springfield, Mass-|Surface |July 16, 1891} 24.1 78 Small Ludlow res., Springfield, Mass.|Bottom.|July 16, 1891} 23.6 71 Tap after passing through 6 miles of PUP ie eek etal GO a iN ct Sie See a aseew ot edpte tala tate July 16, 1891| 23.6 61 Tap after passing through 9 miles of 7) Cr oA STITT SEM LU SR A SY ER WS 3 July 16, 1891] 23.6 57 The amount of dissolved oxygen in ground waters may vary from o to 100 per cent of saturation, and depends upon the char- acter of the material with which the water comes in contact during its passage through the ground. If it meets decaying organic matter at the surface of the ground, or deposits of peat at greater depths, the oxygen may be used up, and especially is this true if iron is present with the organic matter. Table XI gives the amount of dissolved oxygen in certain ground water supplies of Massachusetts. EFFECT OF GASES ON MICROSCOPIC ORGANISMS 12g TABLE XI TABLE SHOWING THE AMOUNT OF DISSOLVED OXYGEN IN CERTAIN DRIVEN WELL WATERS OF MASSACHUSETTS. COMPILED FROM REPT. MASS. ST. BD. HEALTH, 1898, P. 566 TOWN OR CITY (Per sons oP anturabion) HOWE tor o crash orare | 10 INorEht Hastoni ssqaore sees 64 IMairhavent..- sont an oe 47 Kingstony nwa coy avciecses 85 SHarOnmy eo ee eee 93 Malfordyentseracs eerste rae 49 West Brookfield)) 22.252 050: 100 J FehsithoyAeeelhe ae AGehons Obie 43 Sothmhadileyes een ase sel | 100 ASHDUPAHAME ei cciyc ats ela aieye if 100 REVERE sarah ce Hulu scala telenee | 11 Malder an) stream e: | 54 Methuen et yeh SO eas | 9 RGA CIN SAW Func ae cusses doses 100 AMOUNT OF FREE CARBONIC ACID IN NATURAL WATERS UNDER VARIOUS CONDITIONS The carbonic acid dissolved in surface waters is derived partly from the atmosphere and partly from the products of vital activity of organisms living in the water. The amount of carbonic acid in the atmosphere is not a constant quantity. In pure sea air it may be as low as 0.02 per cent (2 parts in 10,000), while in the vicinity of cities it may be 0.05 per cent (5 parts in 10,000). In well-ventilated rooms it should not rise above 0.05 per cent (5 parts in 10,000), but when the ventilation is poor it rises to 0.2 per cent or 0.3 per cent (20 or 30 parts per 10,000), and in crowded rooms where gas is burning it may be even more than this. Out of doors it is generally somewhat higher in summer than in winter, because of the greater activity of organic life. It is usually lower after a rain. The vapor pressure of atmospheric car- bonic acid is not ordinarily sufficient to permit a large amount to be present in solution in water exposed to the air, but as the free car- bonic acid in the water is used up the loss is continually supplied from the atmosphere, so that at times the actual quantity given up by the atmosphere may be very considerable. 130 GEORGE C. WHIPPLE AND HORATIO N. PARKER TABLE XII TABLE SHOWING THE AMOUNT OF FREE CARBONIC ACID IN RAIN WATER (IN PARTS PER MILLION) LOCALITY DATE See REMARKS Brooklyn, N. Y...j/Aug. 1,1901) 6.3 | Latter part of a heavy shower. Brooklyn, N. Y.../Aug. 7,1901/ 2.7 | At end of a long rain. Boston, Mass...... Sept. 18, 1901 6.0 At 1 Ashburton Place. Boston, Mass...... Oct. 14,1901; 8.0 | At 1 Ashburton Place, 11:30 a.m. Boston, Mass...... Oct. 14,1901; 5.0 | At 1 Ashburton Place, 2:45 p.m. Boston, Mass...... Oct. 14,1901; 5.0 | At 1 Ashburton Place, 4:15 p.m. Boston, Mass...... Oct. 15, 1901) 9.0 At 1 Ashburton Place, 9:00 a.m. Boston, Mass...... Oct. 17, 1901} 29.0 | At1 Ashburton Place, after sudden ; shower which lasted 34 hour. Cambridge, Mass../Sept.29,1901/ 5.0 | After three hours’ fall. Fitzwilliam, N. H.|Sept. 1, 1901} 2.5 At beginning of shower, in country. Fitzwilliam, N. H.|/Sept. 1, 1901) 1.8 At end of shower, in country. Rain water contains carbonic acid in amounts corresponding to those present in the atmosphere. In the vicinity of cities it is some- times as high as 20 or 30 parts per million, but where the air is purer it is seldom higher than 3 or 4 parts. Rain at the end of a long storm contains less carbonic acid than at the beginning. This is illustrated by Table No. XII. If rain water is allowed to stand in a shallow receptacle out of doors it will lose much of its carbonic acid in the course of a few hours and the loss will proceed until equi- librium with the carbonic acid of the atmosphere is established. Most of the carbonic acid found in surface waters is not derived directly from the atmosphere by absorption nor by accession of rain- water, but its chief source is to be sought in the living organisms which inhabit the water. All animals exhale carbonic acid, and under certain conditions chlorophyllaceous plants do likewise, but it is chiefly to the bacteria that we look to find this action. These minute plants, countless in number and of enormous aggregate power, are continually breaking down nitrogenous organic matter or ferment- ing carbohydrates, with the consequent liberation of carbonic acid. In many of these vital processes oxygen and carbonic acid appear to act reciprocally: at one time oxygen is taken in and carbonic acid given out, while at another time carbonic acid is used up and oxygen liberated. The production of carbonic acid by bacterial action scarcely needs illustration. The fermentation of the sugars in the absence of oxy- EFFECT OF GASES ON MICROSCOPIC ORGANISMS 131 gen has come to be a common differential test in bacteriological lab- oratories, the observed data including the total amount of gas pro- duced and the percentage which the carbonic acid is of the total gas. Thus in the closed arm of the fermentation tube the common colon bacillus produces enough gas to occupy about 50 per cent of the volume of the tube, and of this about one-third is carbonic acid. Other bacteria produce still greater amounts. As a rule the an- aerobic bacteria produce larger amounts than do the aerobic forms, a fact which accounts for the accumulation of carbonic acid at the bottom of stagnant ponds. Yeasts, molds, and other fungi also produce carbonic acid, and no doubt these play an important part in nature. In general it may be said that wherever decomposition is going on in water there is carbonic acid going into solution, and the larger the amount of organic matter and the more numerous the bacteria the greater is the amount of carbonic acid dissolved. TABLE XIII TABLE SHOWING THE AMOUNT OF FREE CARBONIC ACID IN SURFACE WATERS EXPOSED TO THE ATMOSPHERE (IN PARTS PER MILLION) —-— SOURCE DATE © Aen REMARKS STREAMS OF METROPOLITAN WATER-WORKS, BOSTON, MASS. COLOR Influent to Framingham Res. No. 2) Oct. 23,1901) 6.0 140 Oct. 30,1901) 7.0 126 Nov. 6, 1901) 5.0 86 Nov. 20,1901; 5.2 74 Nov. 26,1901) 5.0 83 Influent to Sudbury Res......... Oct. 23,1901) 5.0 67 Oct. 30,1901) 5.0 54 Nov. 21,1901; 3.6 41 Weenoot, Brookes as seit cs acess: Och Li 190 e820 176 Rock Meadow Brook............. Oct Lie T1901 18.0 148 eT ReliCOWBLOO KN ee4).\ Use. sis elas es: «/5)s Oct. 17, 1901 4.0 130 IBTEW ERE BTOO Kote it sisters sc) a sis ses Oct. 17, 1901) 8.0 82 MOwitay PE LOOKS ial epeleiscatreie.~)la\/ahe els ere Oct. 17,1901) 5.0 69 Brown Meadow Brook............ Oct: L7; 1901), 5.0 49 Cald Sprns Brook. ../.. . «054. ic + Oct. 15,1901) 8.0 145 Oct. 22,1901} 8.0 188 Oct. 29,1901) 8.0 200 Nov. 19, 1901) 5.4 118 Nov. 26, 1901} 6.8 110 ImeiamBrooks cacti Sione elect see Oct. 15, 1901) 19.0 212 Oct. 22,1901; 14.4 204. Oct 29) 1901) 15.0 180 Noy. 12, 1901} 16.0 128 132 GEORGE C. WHIPPLE AND TIORATIO N. PARKER TABLE XIII—-Continued CARBONIC SOURCE DATE NOSED REMARKS STREAMS OF METROPOLITAN WATER-WORKS, BOSTON, MASS. COLOR Indian WBrooky..- pees ence Nov. 19, 1901} 14.6 120 Course BroOkinsigiaiee vai cmometie ee | Oct. (4, CIOL | tONO ya etter Oct TIL gEO 82 Oct. 29,1901) 6.01 94 Pesan Brook. 2 jcjisietse:sieeisyes.s s 5/ep | Get. 4, TOOL!) BeO gia aie ennee Oct OU ao 7 Oct. 29,1901} 4.0 4 Beaver Dam Brook.............:. | Oct) 4, 90H) 710.0) nile iN ee Ocks LL LIOL M920 125 Oct. 29,1901; 6.0 123 Snake Broo kicit ces cys tie caste totes wehowstelete Oct. 29, 1901} 10.0 66 Nashua River, S. Br., Clinton,Mass.| Nov. 8, 1901) 5.0 Lancaster, Mass.| Nov. 8, 1901} 9.2 Us N.Br., Lancaster, Mass.| Nov. 8, 1901} 6.6 Pepperell, Mass.| Nov. 7, 1901} 7.6 OTHER STREAMS. Beaver Brook, Waverly, Mass..... Oct. 13,1901; 5.6 105 Kemp Brook, Fitzwilliam, N. H...| Sept. 6, 1901] 8.0 |APPrO*" 1150 Hempstead Stream, L. I.......... Sept. 5, 1900} 2.0 sell SCHOGACE BOOK. Website's sme Sept. 5, 1900) 5.7 12 Sept. 8, 1900} 3.0 10 Sept. 15, 18 2.5 18 SHALLOW PONDS. Average of 15 shallow ponds on Long Island during summer of DOO tou a Sak eteslns ed teimeels « NOS Ss es eee 3.0 Av. depth about 6 | ft., and ay. color | about 30 Laurel Lake, Fitzwilliam, N. H...| Sept. 6, 1901; 1.5 Boston Public Gardens Pond...... Oce AAS L220 Frog Pond, Boston Common...... Oct. 18,1901; 7.0 | This pond contains Oct. 19,1901) 5.0 a large number of Oct. 21,1901) 5.0 fallen leaves. Oct. 22,1901} 5.0 Oct. 23,1901; 6.0 Oct. 24,1901; 6.0 Oct. 25,1901; 5.0 Oct: (29/1901) 6.0 Farm Pond S. Framingham, Mass.| Nov. 15,1901) 2.0 Nissitisset River, Pepperell, Mass..| Nov. 7, 1901; 6.0 In surface waters which are exposed to the atmosphere and con- tain but little organic matter, the amount of free carbonic acid usually varies from 2 to 5 parts per million. In Brooklyn, N. Y., the average amount in the tap water is 4 parts per million; in Boston it is about 2 or 3 parts per million; in Cambridge, Mass., about 5 parts. In Table XIII will be found the amounts of carbonic acid in various EFFECT OF GASES ON MICROSCOPIC ORGANISMS 133 streams and shallow ponds. It has been found that in a general way the amount of carbonic acid increases with the color of the water, but this is not always the case. Pollution usually increases the amount of carbonic acid present. For example, a stream which flows through the town of Hempstead, L. I., contained on Septem- ber 6, 1900, 2 parts per million above the town and 9 parts per mil- lion below it. TABLE XIV TABLE SHOWING THE AMOUNT OF CARBONIC ACID AT VARIOUS DEPTHS IN DEEP PONDS AND RESERVOIRS CARBONIC POND OR RESERVOIR DATE IN FT, |ACID (parts REMARKS . * |per mill’n) Chestnut Hill Reservoir .|Sept. 20,1901) 1 3.0 13 3.0 26 11.0 Sept. 23, 1901} 1 2.0 13 2.0 26 9.0 Sept. 30,1901; 1 220 13 2.0 25 6.0 Oct) 1571901) 2.0 12 2.0 24 4.0 Ock 2190 1 2.0 |After autumnal over- turning. 12 2.0 24 2.0 Nov. 4, 1901; 1 4.0 2 2.0 24 2.0 Noy. 8, 1901); 1 3.4 2 6.2 24 6.2 Nov. 18,1901; 1 3.0 2 3.0 24 5.0 Hopkinton Res......... Oct. 29,1901} 1 4,0 | After autumnal over- turning. 26 4.0 52 4.0 Framingham Res. No. 3./Nov. 15,1901} 1 3.0 WHGDUGY RES. to... 26 6 Nov. 15, 1901 it 4.0 3) 2 aa) £65700 |r Dec. 6,1901; 1 1.4 12 ie0 24 8.0 HetisrRies|}G:ELe ....5. sei Dee Ge LOOM rrr 3.6 Fresh Pond, Cambridge../Sept. 28,1901; 1 1.0 23 8.0 46 16.0 134 GEORGE C. WHIPPLE AND HORATIO N. PARKER The amount of dissolved carbonic acid found in a shallow surface water depends, however, upon other factors than those which cause its formation. It is always complicated by reason of its reaction with the normal carbonates of the alkaline earths with which it may come in contact, and there is usually a tendency for it to diffuse into the atmosphere. This diffusion depends upon the vapor pressure of the atmospheric carbonic acid and the extent to which the water is exposed to the air. In the middle of swamps which are covered with tall vegetation to such an extent that the air does not circulate freely the carbonic acid is often high in the air and in the water in contact with it. At the bottom of thermally stratified ponds there is no opportunity for the water to come in contact with the atmos- phere, and consequently there is often a concentration of carbonic acid in the lower strata. This is especially true if there are deposits of mud and organic matter at the bottom. (See tables XIV, XV, and XVI.) TABLE XV TABLE SHOWING THE AMOUNT OF FREE CARBONIC ACID AT DIFFERENT DEPTHS IN LAKE COCHITUATE (IN PARTS PER MILLION) peptH| 1900 | 1900 | 1900%| 1901 | 1901 | 1901 | 1901 | 1901 | 1901 1901 2° INFT. |ocr. 12] Nov. 9 |Nov. 16) May 24|JUNE 21/JULY 19/auG. 28) ocT.11| Nov. 8| Nov. 14 1 2.00) 22847 83.6) 2.6) 92.0) 92.0") 2.0 aoa eee 5 SED |S eb abe tereatel teysist cdots SA Ase BERGE bee concen olistas sooo on 10 ALO PAOUl| seals 2.0} 5.634) 3205) 2: 0a SON eae Onl erertererteee 15 pa 3) ee al See 5.25.6 | 620)) 5./0)) (400 a On eee 20 dirs 0) ate al eA Ane 6:8 | (6.04) 620) |) "G.08) 1OLOn e420 n eee Pigiay) a! A a fs ee ee 6.0) |G Oin ae 6.05). cee 6.0(26 ft.) 30 OES PAO nie cise = 6.0 | 6.0) 6.0 | G.0)) 11.03) 40h eee ee 35 Sa 5) M9 a ee ee AS RPE me EMH ASH Sr ilacok ot cons 4050 OU Loz Beles cer 10.0'| ‘6.0 1. 6.0))' \7.0)) ELI00 > Tare ee Ca Yi ie Beato 59 ft ho Jars Pe ea sel peer el Ineiceel loecooolidae con |saocaclsccocs or o- 50 || 17.50) 16.6 | 3.7 | 8.0) 9.6:)' 8.0) 8.0) 19:0) 2E Oe eee BSE ssa civetcilioes aaa ey oa EA Seoul gla the 10.0} 16.0" | 23.0") eee 6.0(52 ft.) 60 | ole cnots 2.0) 16.0 | 23:0 | 24500) i-sceecee The phenomena which take place at the bottom of a thermally stratified lake are most interesting. Below the thermocline the water is shut off from communication with the atmosphere. The processes of vital activity soon cause a diminution in the amount of dissolved oxygen and an increase in the amount of free carbonic 9 The lake overturned on Nov. 12, 1900. 10 The lake overturned on Nov. 10, 1901. EFFECT OF GASES ON MICROSCOPIC ORGANISMS 135 acid. If there is a deposit of organic matter at the bottom the oxygen may become entirely exhausted, after which decomposi- tion may occur under anaerobic conditions. The anaerobic bacteria are vigorous producers of carbonic acid, and consequently a large amount of this gas goes into solution. The carbonic acid also unites with the iron present at the bottom, forming soluble ferrous car- bonate. This gives rise to an increased color and turbidity of the water, which, on exposure of the water to the air, is still further increased by the oxidation and precipitation of the iron. Under the conditions which thus prevail at the bottom of a stagnant lake Cren- othrix may develop. After the period of stratification ceases and circulation takes place through the vertical, the products of decompo- sition become distributed through the entire body of water. These phenomena are well illustrated by observations made at Lake Cochituate, and given in Table XV. See also Plates XXI and XXII. TABLE XVI TABLE SHOWING THE AMOUNT OF CARBONIC ACID AT VARIOUS DEPTHS IN A SMALL DISTRIBUTION RESERVOIR IN WHICH GROUND WATER IS STORED, TOGETHER WITH THE TEMPERATURE AND THE NUMBER OF MICROSCOPIC ORGANISMS PER C.C. SEPTEMBER 4, 1900 JULY 25, 1901 s Amt. of free No. of micro-|| = Amt. of free No. of micro- @. |Temperature|carbonic acid > "|| @ |Temperature|carbonic acid F i B|'Gair) fim parts perfsobicorgen:| 2 )"“Cahr.) fin parts perigee Ore, 0 80.0 ORO Pieters ais 0 79.5 —5.0 5760 2 79.5 0.0 140 2 79.5 POF Miesinisis, ove ioreneds 4 79.5 QP rear sists eave i 79.0 SOU SH aioe siete 6 79.5 HO Aira ° “30 Caotieas saauese PLATE XxX Be tor ger ae = i?) “NeITIIWw use sitvd 3 3 | = ~ ° A ES Se fee) | | eee mee | | eee JAN IN THE AND AT FROM ANALYSES BY DIAGRAM SHOWING THE AMOUNT OF DISSOLVED OXYGEN DIAGRAM SHOW)NG THE AVERAGE TEMPERATURE OF THE WATER OF THE RIVER SEINE AT CHOISY-LE- ROI IN BOSTON,MASS. AND THE CORRESPONDING TAP WATER 1898. ‘M. ALBERT- LEVY. THE BROKEN LINE REPRESENTS SATURATION ARGENTEUIL DURING THE YEAR is WATER OXYGEN WHEN THE AmounT OF pissolveo SATURATED, AT THE OBSERVED TEMPERATURES. PLATE XXI @06/ 9/ Ao, /06/°9/ 13D ILYNLIHIOD FHYI NI SHASTA SNOIWYAN LY FLYIDAHAONYAD SO YIPAWNAN FHL ONY F190 DWNOBYED FIHAS SO LNIOWY FHL SILOM FHL IO FYMLYYFTANIL FHL ONIMOHS WHHIY/ ones, o/c07905 ee, SUL SAS OU MAUD) (oon A OMMINOE!, i bad Pe Peas YNIGEYNY -—— —— — PVIDANASON VLD FVLOL ———— FYIDPAHAONGAD /osig2 one 406/61 ATA 406/18 INA, ot lyn —— 3 008/ 009! 00%! 0! 00/1 warn tad 21802 ad SLIT auwbaNYLE FO wraway 0190 DINOBEYD ” ” IENLGYYTAN FL HLGIO 4577 N/ PLATE XXII | ‘99 KIS SYNOWO??eW ¥O YIBWIAWV o9 0os/ ° FLvO Iwes FHL NO SHLASIT SNOIWHA Lv YILEYM FHL IO FLNLYSIASN IL FHL ONY ‘968/121 ATAL NO PLYONLIA DOD FIWY7 NI SYNOWO?IIVGW SO NOVLNG/YLSIO WIILYIA FHL ONIMONS Wrbydv/a FANLSAIAVIL Diagram showing the amount of Crenothrix 00§ v0t standard (in ‘2°29 Hid SIAWIN 005 in units) at various depths “summer of 1901. SS bw S ly RX LS) % SEE IY, Spf GefENG S N ee THE STRUCTURE AND CLASSIFICATION OF THE CONJUGATAE WITH A REVISION OF THE FAMILIES AND A REARRANGEMENT OF THE NORTH AMERICAN GENERA By CHARLES E. BESSEY, PuH.D. The preceding papers on Diatoms! and Desmids? have prepared the way for a discussion of the structure and classification of the remaining family (Zygnemataceae) as well as of the whole order (Conjugatae). In the papers cited it has been assumed that the diatoms and desmids have descended from filamentous ancestors, and these plants are there regarded as still typically filamentous, the filaments in most cases undergoing early solution. This view does not regard the diatoms and desmids as properly unicellular plants, although the cells of these organisms are for the most part early isolated, and pass their lives in this condition. The Pond Scums (Zygnemataceae) constitute a third family of the order Conjugatae, and here, while the filamentous structure 1s preserved, the adhesion of cell to cell is so feeble that fracture takes place very easily. Although the cells do not separate so as to exist singly, there is evidence that spontaneous fragmentation does occur, the result being the formation of short, few-celled filaments, comparable to the “hormogones” of the Cyanophyceae. This tend- ency to individuality in the cells is indicated by the fact that in ad- jacent cells one may be dead and the other in vigorous life, or one may have taken part in the formation of a zygote by a sexual act, while the other is still active vegetatively. It may be assumed that these plants have been derived from other filamentous forms, and that the adhesion of cell to cell and the con- sequent formation of a multicellular plant body had become a well- ~ 1"“The modern conception cf the structure and classification of Diatoms,” in Transactions of the American Microscopical Society, Vol. XXI, p. 61. 2‘The modern conception of the structure and classification of Desmids,” in Transactions of the American Microscopical Society, Vol. XXII, p. 98. 146 CHARLES E. BESSEY established habit long before the peculiarities arose which set them off as Zygnemataceae. Now in any existing plants which may have given rise to the Zygnemataceae the individuality of the cells is distinctly subordinated to the individuality of the filament, and there is a much closer relation of cell to cell. Such groups as the Ulotrichaceae and Chaetophoraceae, with their highly individualized filaments, may have given rise to the Zygnemataceae, and in this paper I shall assume such relationship for purposes of comparison. In the families named the filaments are composed of proper cells (uninucleate), and the chromatophores, which are relatively large and few in number, vary from a single broad band which encir- cles the nucleus, to several narrow, longitudinal bands. The differ- ences between these cells and those of the Zygnemataceae are little if any greater than those to be found within the limits of the fam- ilies, and it is reasonable to suppose that the necessary structural changes may have occurred as here suggested. In the Ulotrichaceae and Chaetophoraceae the plants are propa- gated mainly by zoospores, and also by the fragmentation of the filaments artificially, and possibly spontaneously. In the Zygne- mataceae fragmentation has been much increased, and propagation by means of zoospores quite suppressed, possibly because of the fact that these plants float upon still waters where fragmentation alone provides amply for their propagation. Motile zoospores being thus quite needless have accordingly disappeared. The quiet habitat of the Zygnemataceae doubtless had to do, also, with the change in the motility of the gametes. While in the other families mentioned the gametes are ciliated and actively motile, as indeed is quite necessary in the running waters which they inhabit, in Zygnemataceae no such activity is necessary; and here the slug- gish gametes, no longer ciliated, are guided in their short journey by the device of tubular extensions of their cell walls. The whole process in these plants is a much more sluggish one than in the plants with which they are here compared. There is thus a marked degeneration in the filament of the Zygne- mataceae, which shows itself in its ready fragmentation, and in the greater sluggishness of the gametes in generation. Both of these tendencies receive greater emphasis in the Desmids and Diatoms where fragmentation is so marked that in most genera the cells are isolated for the greater part of their existence. It appears also — STRUCTURE AND CLASSIFICATION OF THE CONJ UGATAE 147 that the sluggishness in generation in the Zygnemataceae has been followed in some of the Desmids and Diatoms by a partial, if not complete, suppression of the sexual act. According to this view “conjugation,” as the sexual act in the Conjugatae has been aptly called, is the result of degeneration. It is sexual reproduction on its way toward disappearance. Instead of affording an example of the beginning of sexuality, as has so often been suggested, these plants show sexuality on its way to disappearance. These conceptions of the nature of these plants, and their rela- tionship to other algae, require a considerable rearrangement in the sequence of the families and genera. In the papers cited above I have given expression to my ideas as to the proper sequence of the tribes and genera of the Desmids and Diatoms. The Zygnemataceae constitute so small a group that the task of arranging the genera in accordance with these ideas is not difficult. I have followed De Toni? in including the Mesocarpeae, which in my opinion do not -differ sufficiently to be set off as a separate family. Wille,* on the contrary, regards the latter as entitled to family rank under the title Mesocarpaceae. The order Conjugatae as here understood may be briefly char- acterized as follows: Order CONJUGATAE Plants microscopic, consisting typically of simple, unbranched rows of cells, often separating early into isolated cells; green, with lamelliform, taeniform (ribbon-like), or granular chromatophores, in one family yellowish by the addition of phycoxanthin ; propaga- tion by cell fission; generation by the union of the protoplasm of pairs of cells (conjugation, or aplanatic isogamy). KEY TO THE FAMILIES. A. Cells in cylindrical filaments 1. Zygnemataceae. B. Cells mostly solitary, with cellulose walls 2. Desmidiaceae. C. Cells mostly solitary, with siliceous walls 3. Bacillariaceae. 3Sylloge Algarum, by J. B. De Toni, p. 710. 1889. 4Die Natiirlichen Pflanzenfamilien, by A. Engler and K. Prantl, Vol.I, Abt. 2. 1890. 4 148 CHARLES E. BESSEY Family 1. ZyYGNEMATACEAE (“Pond Scums”’) Cells with thin, smooth, cellulose walls, green, cylindrical, never constricted, always united into unbranched filaments; propagation by the accidental or spontaneous breaking of the filaments into short segments which grow directly into new plants; in some cases resting cells (aplanospores) are formed by a condensation of the cell contents and a thickening of the wall, these growing later into new plants; generation by the growth toward one another of tubular protrusions from adjacent or approximate cells and the formation of a continuous tube by the absorption of the contiguous end walls; through this tube the whole or part of the protoplasms of the two cells unite, either in one of the cell cavities or in the tube itself, forming a thick-walled resting-spore (zygote), which eventually grows directly into a new filament.—Minute fresh-water plants, floating on the surfaces of quiet pools and ponds. Key TO THE GENERA. A. Whole contents of the conjugating cells entering the zygote. I. Conjugating tubes not septate. a. Chromatophores I or more, parietal, taeniform, spiral. 1. Conjugata. b. Chromatophores 2, stellate, axial, 2. Lucernaria. c. Chromatophore a single axial plate, 3. Debarya. II. Each conjugating tube with a septum at its base, chro- matophores 2, stellate, 4. Zygogonwm. B. Part of the contents of the conjugating cells entering the zygote. I. Chromatophore a single axial plate, aplanospores none, 5. Serpentinaria. II. Chromatophore a single axial plate, aplanospores present, 6. Gonatonema. 1. Conjugata Vaucher (Spirogyra Link).°—Cells three to ten times longer than broad (rarely only as long as broad) ; transverse walls plane or with circular folds; chromatophores one or more, taeniform, parietal, spiral, each with several pyrenoids; nucleus centrally suspended; conjugation between two cells of different fila- 5 Unfortunately the earlier name Conjugata, applied to these plants by Vaucher in 1803, must replace the well-known Spirogyra of Link, dating from 1820. STRUCTURE AND CLASSIFICATION OF THE CONJ UGATAE 149 ments or of the same filament ; zygote formed in one of the conju- gating cells—Species many, in fresh waters. 2. Lucernaria Roussel (Zygnema of authors).°—Cells as long as, or two to five times as long as broad; transverse walls plane; chro- matophores two, axial, stellate, each with one pyrenoid; nucleus central, between the chromatophores; conjugation between two cells of different filaments or of the same filament; zygote formed in one of the conjugating cells, or in the conjugation tube.—Species many, in fresh waters. 3. Dabarya Wittrock.—Cells five to ten times as long as broad; transverse walls plane; chromatophore a single axial plate; conju- gation between two cells of different filaments; zygote formed in the conjugation tube——Species one, in fresh waters. 4. Zygogonium (Kiitzing) De Bary.—Cells from shorter than broad to twice as long; transverse walls plane ; chromatophores two, axial, irregular (sometimes joined in an axial strand) ; conjuga- tion between two cells of different filaments, the two protoplasms in the conjugation tube cut off from their cells by partitions before union; zygote formed in the conjugation tube.—Species two, in fresh water. 5. Serpentinaria S. F. Gray (Mougeotia Agardh).’—Cells many times as long as broad; transverse walls lenticular ; chromatophore one, axial, lamelliform, with two or more pyrenoids; conjugation between two cells of different filaments, or of the same filament, only a part of the protoplasm of each cell uniting to form the zygote, which lies in the conjugation tube and is separated from the cells by partitions—Species many, in fresh waters. 6. Gonatonema Wittrock.—Vegetative cells as in Serpentinaria; conjugation unknown; non-sexual spores (aplanospores) produced by the elongation of cells and the formation of a pair of partitions near the middle to separate the zygote-like cells—Species two, in fresh waters. 6 Here again it is unfortunate that we are obliged to displace the familiar name Zygnema of S. F. Gray and Agardh (1821 and 1824) for the earlier name Lucernaria proposed by Roussel in 1806. 7Mougeotia dates from 1824, when it was proposed by Agardh in his Systema Algarum, but this is antedated by Sevfentinaria proposed by S. F. Gray in his Natural Arrangement of Brit- ish Plants, published in 1821. 10 I50 CHARLES E. BESSEY Family 2. DesmipracearE (‘‘Desmids”) See “The modern conception of the structure and classification of Desmids” in the Transactions of the American Microscopical Society, Vol. XXII, pp. 89 to 96, for a revision of the tribes and a rearrangement of the North American genera. Family 3. BAcILLARIACEAE (‘‘Diatoms’’) See “The modern conception of the structure and classification of Diatoms” in the Transactions of the American Microscopical Society, Vol. XXI, pp. 61 to 85, for a revision of the tribes and a rearrangement of the North American genera, ON HYMENOLEPIS CARIOCA (MAGALHAES) AND H. MEGALOPS (NITZSCH) WITH REMARKS ON THE CLASSIFICATION OF THE GROUP. By B. H. RANSOM WITH THREE PLATES Hymenolepis carioca (Magalhaes) One of the most common tapeworms of chickens in Nebraska, Iowa, and Missouri is the form noted by Stiles (96, p. 59) under the title of Taenia sp. Conard MS. Through the kindness of Dr. Conard I have had the privilege of seeing his manuscript, as well as his material and preparations, which have proved of great assist- ance in the identification of my specimens. The worm in question is very slender and delicate. Scarcely larger than a coarse thread at its posterior end, it tapers gradually anteriad, becoming exceedingly tenuous at the neck. The length ranges from 30 mm. to 80 mm. The width at the neck varies be- tween 75 « and 150 p, at the posterior end from 500» to 700 pw. The margins of the strobila are serrate (fig. 9), although the appear- ance of serration is often obliterated when the segments are much expanded (fig. 1). The posterior margins are not prolonged back- ward to any extent, so that there is little or no overlapping of the segments. Throughout the strobila the width of the segments is three to five times, or more, greater than their length; the worm is thus of the type with short segments such as Hymenolepis diminuta and others of the genus. The head (fig. 8) is small, somewhat flattened dorso-ventrally. It measures from 140 » to 160 pw in length, 150 w to 215 p in width, and 100 » to 140 » in thickness. The suckers are shallow and slightly oval, with a diameter of 70 # to go uw. The rostellum (fig. 8, ros) is like that of Hymenolepis diminuta. It measures as it 152 B. H. RANSOM lies withdrawn into the head, 25 » to 40 m in diameter by go » to 100 » in length. As in H. diminuta, there is a small pocket (1p), lined with cuticula, and opening at its tip. The rostellum is un- armed. A head, sectioned im situ with a piece of intestine, pos- sessed, upon the suckers, hooks (fig. 3) which were very small and caducous. The ventral root is long, while the dorsal root is only a mere knob. The slender, pointed blade forms an angle of go° to 120° with the ventral root. They are of the same type as the hooks from the suckers of Davainea Friedbergeri (Stiles 96, fig. 237). The smallest have a blade 4 or 5 in length, and a ventral root of 2pto3 om. The blade of the largest hooks measures 6 » to 7.5 p, the ventral root 2» to4 ». The round neck measures from 0.6 mm. to 1.5 mm. in length, and 0.075 mm. to 0.15 mm. in width. The genital pores are situated upon the right-hand margin of the strobila, normally somewhat in front of the middle of each seg- ment (fig. I, gc). Very rarely a pore will be found on the left-hand margin. The cirrus pouch (cp), seminal vesicle (vs), and seminal receptacle (sr) are usually easily recognizable in toto specimens. In ripe segments the last (fig. 9, sr) is quite apparent by reason of the mass of spermatozoa it contains. The last 30 to 100 segments are crowded with embryos so that the median field of each is fully oc- cupied (fig. 9), and the embryos of one segment are separated from those of the adjacent one only by a double thickness of the thin uterine wall. Since the segments do not tend to break off singly from the strobila as they become ripe, and since the embryos which they contain form practically a continuous mass extending unbroken from one proglottis to the next, if a portion of the worm be broken off from its posterior end the entire series of proglottides will con- stitute what is in effect but a single embryo sac. Four membranes may be distinguished surrounding the oncho- sphere (fig. 6). The space between the middle two is filled with a granular mass, and potentially, as well as actually in some cases, these two membranes, by close approximation, constitute but a single envelope. The three outer layers are thin and colorless, while the innermost next the embryo is often slightly tinged with yellow and is usually thicker than the others. ON HYMENOLEPIS CARIOCA AND H. MEGALOPS 153 MEASUREMENTS OF EMBRYOS SIZES IN MICRONS LEAST GREATEST Dee eReNWEIOPe Lard 2 'sots fate oie ates fdatete sare ae 36 x 36 70 x 75 ter middie envelope. 0. <2... sclnasds 520 nos 30 x 30 65 x 60 HEMEE THIAGIC eVELOPEe. .!yi.5 <0 ase sally oss aes 26 x 26 40 x 35 ERAS TACTIVCL ODES d/afel otaicioiale s crefeteleketeraraie|apstah etaiets\ 24x 16 29 x 21 PREIS HELO Cc chan cS 545 shat latalnonm a aeetel etre 18 x 14 27x19 1313.0) 35) (Gi) ees SODA AHO bone Obionqaacoe 10 12 A comparison of the points given above with the description and figures by Magalhaes (98) of Davainea(?) carioca will show that the Taenia sp? of Conard is the same as the form which Magalhaes has described. The presence of hooks upon the suckers might ap- pear as a confirmation of the supposition made by Magalhaes that the species is a Davainea, but beyond this characteristic it possesses none of the peculiarities of that genus. The fact alone that it has a definite and persistent uterus precludes the possibility of such a generic relationship, and furthermore its close structural resem- blances to the type of Hymenolepis justify its immediate reference to that genus. The possession of armed suckers by a species of Hymenolepis is significant in that it demonstrates how little importance can be attached to such a character in establishing generic relationships. In fact, in many cases it can not be considered of more than specific value. Braun (94-00, p. 1718) has gone so far as to assign to this character a rank even higher than generic, by making it the sole distinguishing mark of the subfamily Davaineinae, comprising the genera Davainea, Echinocotyle, and Ophryocotyle. He has thus bound together three groups of forms resembling each other only in the possession of armed suckers, and at the same time has sep- arated the group Davainea from other forms which bear close re- semblances to it in many respects, but do not have hooks upon the suckers. Beyond the fact that Echinocotyle and Ophryocotyle have armed suckers there can be no excuse, so far as our knowledge of their anatomy goes at present, for grouping them with Davainea. Moreover, such genera as Monopylidium and Cotugnia, although their suckers are unarmed, resemble Davainea too much to be placed in a separate subfamily, and certainly have more in common with 154 B. H. RANSOM the latter than with Hymenolepis or Choanotaenia. The subfamily Davaeneinae consequently can no longer be maintained upon its original basis. INTERNAL ANATOMY.—WNervous System.—Owing to the small size of the worm only a few of the details of the nervous system could be determined. There is, however, a well-defined bilobed ganglionic mass (fig. 10, cg) situated at the base of the rostellum. From it nervous processes (au) pass anteriad along the sides of the rostellum, within which, also, there is a considerable amount of nervous tissue. The lateral longitudinal nerves (Jn) arise from the postero-lateral corners of the ganglia (figs. 1, 7, Jn). The accessory lateral nerves are very small and only occasionally evident in the segments, lying a short distance dorsal and ventral from the main nerves. Musculature—Extending throughout the strobila are four dor- sal and four ventral muscle strands, or small compact bundles, com- posed of only a few fibers (fig. 7, im). In the scolex they attach to the suckers, two to each. These slender strings comprise the inner longitudinal muscle layer, and correspond to the eight inner longitudinal muscle bundles of Taenia inflata Rud. as described by Jacobi (98) and of Taenia microsoma, Drepanidotaenia anatina, and another undetermined species as described by Wolffhtigel (00). The outer layer of longitudinal muscles (om) consists of about one hundred muscle strands similar to the inner muscles and like them continuous from segment to segment. The origin of these muscles is similar to that of the outer longitudinal muscles of Anoplo- cephala perfoliata as described by Lithe (94, 96), 1. e., they are the prolongations of the longitudinal subcuticular muscles of the scolex. A layer of diagonal muscles (fig. 7, dm) is prominent in the older segments just outside the outer longitudinal layer. A transverse muscle system is represented only by a few slender iso- lated fibers (tm), restricted mostly to the extreme anterior and posterior ends of the segment, as is the case in many other forms of the genus. A few dorso-ventral fibers are also found in these regions. The sac-like rostellum (fig. 10, ros) possesses a muscular wall consisting of an outer layer of longitudinal and an inner layer of circular fibers, as in H. diminuta (Zschokke 88). Excretory System.—The longitudinal excretory canals unite in ON HYMENOLEPIS CARIOCA AND H. MEGALOPS 155 the scolex to form a ring at the base of the rostellum. From this ring four small vessels, about 1 » in diameter with comparatively thick, highly refractive walls, enter the rostellum a short distance in front of its base. Near the middle of the rostellum the dorsal vessel on each side unites with the ventral vessel so that two closed loops are formed. Branches of the excretory system apparently sim- ilar to these loops have been described by Mingazzini (99) for Hymenolepis murina. I have also found exactly similar loops in the rostellum of H. diminuta, readily apparent in cross-sections. The ventral excretory vessels are larger than the dorsal vessels from the beginning (fig. 10, vc, dc). In the scolex the former measure 6 p, the latter 4 ». The ventral vessels later attain a size of 25 » to 40 »; the dorsal vessels retain their original small diameter or be- come even more attenuated. Transverse vessels connect the ventral vessels at irregular intervals of two to seven segments, usually at an interval of about five. Reproductive Organs.—The three testes (figs. I, 2, 7, ¢) lie near the middle of the segment and usually two to the left and one to the right of the median line. The vasa efferentia (fig. 2, ve) unite near the median line to form the vas deferens (vd) which runs forward to a point dorsal to the inner end of the cirrus pouch. It here turns to the left, then curves ventrad, and finally, bending to the right, enters the base of the latter. Between the first and second turns of the vas deferens, dorsal to the cirrus pouch, there is an enlargement, the vesicula seminalis (figs. 1, 7, vs, fig. 2, vsa), which may attain a size of 50 by 70 »& §©Scattered over its surface, more especially at its distal end, are numerous large cells drawn out at one pole into a narrow process (fig. 7, pc). The body of one of these cells is some 6 » or 8 p» in diameter, while the length of the process, which has a diameter of about I p, is 12 ¢to 14 pw. The protoplasm is compact and finely granular, and each cell contains a spherical nucleus 2 » in diameter, with a single prominent nucleolus. These cells are to all appearances exactly similar to the cells found in the same position but in much greater numbers in Taenia transversaria, Taenia expansa, and Calliobothri- um coronatum, described by Zschokke (88) as prostate glands. I was unable to demonstrate, however, that these cells opened into the cavity of the vas deferens in H. carioca as described and fig- ured by Zschokke. In the wall of the vas deferens between the 156 B. H. RANSOM vesicula and the cirrus pouch a few circular fibers are sometimes evident (fig. 7). The cirrus pouch (fig. I, cp) in adult segments measures from 120 » to 175 pw in length by 15 »to 18 win diameter. It is almost cylindrical, rarely perfectly straight, but bent more or less, usually toward the ventral surface in a gentle curve (fig. 7). Along its outer surface there are about twenty prominent longitud- inal muscle bands, 2 » to 3 » in thickness and slightly less in width (fig. 4, mp). Similar muscle bands have been described in Taenia depressa by Fuhrmann (95), in Taenia inflata by Jacobi (98), in Fimbriaria fasciolaris and Dicranotaenia coronula by Wolfthiigel (00), as well as in other forms by different authors. Hymenolepis carioca is remarkable by reason of the small number of these bands, and their relatively large size. Surrounding the middle part of the cirrus pouch is a layer of cells (figs. 4, 7, my), which, from their inti- mate relation to the muscle bands, are to be interpreted as myoblasts. They possess nuclei 3 » in diameter, each containing a deeply stain- ing nucleolus. These myoblasts are most prominent in the young segments before spermatozoa are found in the vas deferens. Cir- cular muscles are lacking, and the membrane of the cirrus pouch (fig. 4, sm) lies directly beneath the muscle plates. Upon entering the cirrus pouch the vas deferens is much constricted (fig. 7). In thickness and general appearance, the wall of this narrow portion of the vas deferens resembles the membrane of the pouch, which seems to have turned in at this point to form a narrow tube through which the vas deferens passes. Beyond this narrow portion the vas deferens is dilated to form a second seminal vesicle (figs. 4, 7, vs’) The wall of this part of the vas deferens is surrounded by circular fibers which are most prominent in the region occupying the mid- dle third of the pouch (fig. 7). At a point about one-third the length of the pouch from its distal end, the vas deferens becomes narrow again, to form the cirrus (fig. 7, ci), a thin tube not more than I » in diameter, without apparent spines or musculature. In the space surrounding the vas deferens within the cirrus pouch are numerous small nuclei, I # to 2 w in diameter (figs. 4, 7). Both the cirrus pouch and the vagina are dorsal to the nerve and the ex- cretory canal. From the diagonal muscles of the proglottis fibers turn in to attach to the outer portion of the cirrus sac, serving thus as protractors (fig. 7, pr). ON HYMENOLEPIS CARIOCA AND H. MEGALOPS 157 The genital cloaca (fig. 1, gc, fig. 7) is from 12 » to 36 mw deep, sur- rounded by longitudinal and circular fibers, the latter next the cuticula. The longitudinal fibers come from the diagonal system, or from the subcuticular longitudinal muscles, and attach to the tip of the cirrus pouch to form part of the system of protractors. The opening of the vagina into the cloaca is ventral and posterior with respect to the cirrus. The vagina (figs. 1, 7, vg) is at first narrow, being only 1 » in diameter. At a distance of 8» to IO p from its distal end, it is surrounded by a small bulb-like body (fig. 7,vUgs), consisting of short, thick, muscular fibers running in a spiral. This bulb is apparently homologous to the vaginal sphincter of Drepanidotaenia lanceolata described by Wolffhiigel (00a). Be- yond the sphincter the vagina gradually enlarges, and swells out into a seminal receptacle, which may grow to be very large, so as to reach forward to the anterior limits of the segment, and even crowd in against the organs of the next. Inward it may extend consider- ably beyond the proximal end of the cirrus sac. The ovary (fig. 1, ov) is a sac-like organ elongated transversely, faintly bilobed, or even slightly trilobed, as in Taenia inflata (Jacobi 98), lying in the posterior half of the proglottis toward its ventral surface. At its maximum it extends from one excretory canal to the other. The ova reach a diameter of Io p» before leav- ing the ovary. The yolk gland (yg) is spherical or ovoid, 30 » to 40 » in diam- eter, situated posterior and dorsal to the ovary, near the median | line. The uterus at first is simply a solid cord of cells (fig. 1, ut) ex- tending transversely along the anterior border of the ovary and reaching the excretory canals on either side. With progressing de- velopment a cavity is formed by a hollowing out of the cord, and the uterus becomes a thin-walled sac which grows backward on the dorsal side of the ovary. As the uterus enlarges the ovary quickly disappears, and the former soon comes to occupy all the available space within the proglottis (fig. 9). The wall of the uterus con- sists of a thin membrane (fig. 7, uf) upon the outer surface of which are numerous small cells elongated sagittally, and fine fibers which seem to be extensions of the pointed ends of the cells. During the growth of the uterus a number of infoldings arise in its wall, mostly in the form of tubular processes, a few of which meet and fuse to 158 B. H. RANSOM form slender, bridge-like connections, so that finally the uterus is: modified to a slight degree from its original simple sac-like condi- tion. With regard to its reproductive organs as well as in other respects. Hymenolepis carioca is quite comparable to H. diminuta (Rud.), the type of the genus. Both forms possess unilateral genital pores, reproductive canals dorsal to the nerve and excretory canals, three testes, a large sac-like vesicula outside the cirrus pouch, and a sec- ond smaller one within. A large seminal receptacle, a more or less bilobed ovary, and a yolk gland posterior and dorsal to the latter are also present in both. The uterus in both arises first as a trans- verse tube anterior to the ovary, later filling the segment, and in each case is not a simple sac, but characterized by a greater or less number of inturned processes and bridges of tissue developed from its walls. Apart from the major complications arising during the growth of the uterus of H. diminuta as described by Zschokke (88), there are a great number of small processes extending from the wall of the uterus in among the eggs, which are quite like the com- paratively insignificant infoldings of the wall of the uterus in H. carioca. The similarity between the eggs is obvious. Hymenolepis megalops (Nitzsch) Was first found by Nitzsch in the intestine of Anas boschas and Dafila acuta. Creplin (25) found in the rectum of Anas marila a specimen 54 mm. long and 2.25 mm. wide posteriorly, possess- ing a head almost 2.25 mm. in width, with a very short, obtuse rostellum. Later he found five much smaller specimens of which he gave incomplete descriptions. Two figures by Creplin (29) are reproduced by Braun (94-00). Dujardin (45) took two specimens from an Anas boschas, one of which was 52 mm., the other 35 mm. in length. He gives two figures of detached proglot- tides, and in addition a diagnosis of the form upon the basis of its external anatomy. Diesing (50, 64) merely adds to the list of hosts. Stiles (96) gives a short synopsis of Taenia megalops from previous authors, with drawings of a specimen taken from Anas braziliensis from the collection of the Vienna Museum. While he is of the opinion that from existing descriptions the worm can not be recognized with any degree of certainty, I believe, on the other ON HYMENOLEPIS CARIOCA AND H. MEGALOPS 150 hand, that the external characters are peculiar enough to enable one to identify it easily, and with little or no chance of error. Four individuals of this tapeworm were collected by me, March, 1901, from the rectum of a pintail duck, Dafila acuta, shot on the Missouri river near Columbia, Mo. Two of the worms were young and immature. The other two were larger, and considerably more advanced in development, as several of the posterior segments con- tained embryos which were yet, however, without hooks. The length of the largest specimen was 35 mm., and the width, almost uniform from head to tail, 0.55 mm. just behind the head, and 0.72 mm. at a point near the posterior end. The number of segments was 195. The head is large, 1 mm. in length by 1.1 mm. in width and thickness, and, viewed from the front, square with rounded angles (figs. 11, 17). The suckers are spherical, 0.4 mm. in diameter, and open forwards and outwards, near the four angles of the flattened anterior surface of the head. In the center of the anterior surface is the small orfice of the cavity of the rostellum. The length of one of the small specimens was 4 mm., its width 0.45 mm., and the number of segments about 70. The head measured 0.45 mm. in length by 0.75 mm. in width and thickness. Strobilation begins immediately behind the head, and is very distinct almost from the first, by reason of the deep constrictions between the segments. These constrictions mark off a peripheral portion of the segment from the central portion (figs. 18, 19, 20). The central portions are continuous, while the peripheral portions are separated from one another by the constrictions. The outer surface of the projecting rims of the segments is marked by a number of longitudinal grooves (fig. 20, gr) noticed by Dujardin (45), which give the surface of the worm a corrugated appearance. The under surface of the rims is marked by tiny longitudinal ridges in the cuticula, folds made necessary by the decrease in area of the under surface when the rims, instead of extending laterally as in the youngest segments, come to point backward, as they do more and more with the increasing age of the segments. 160 B. H. RANSOM MEASUREMENTS 25TH PR. | 175TH PR. Width of proglottis from edge to edge of rim.............. 550 » 720 » Width of central portion of proglottis................000- 350 4 440 Length of central portion of proglottis................000. 30 & 290 & Length ofentire proglottis.). 2:5 hi4.- Sac nace oeaeiee see 30 490 & Thickness of proglottis from edge to edge of rim.......... 400 & 280 Thickness of central portion of proglottis................. 200 » 240 » The genital pores (figs. 15, 19, gc) are unilateral, situated on the right-hand margin, and marked by a slight prominence. A line drawn transversely through the very base of one segment will pass through the margin of the preceding segment very near its genital pore. The main body of the proglottis thus lies anterior to the genital pore, with only the projecting rim extended behind. Through Dr. Ward I am indebted to Dr. C. W. Stiles for the op- portunity of comparing with my specimens one from the U. S. Na- tional Museum, the same from which the figures by Stiles (96) were drawn. This worm was much contracted and considerably shorter than my largest specimen, but possessed a few more seg- ments, 200 to 210 in all, the posterior ones of which were transpar- ent enough to show that they contained six-hooked embryos. The embryos at the latest stage (fig. 13) reached in my speci- mens are round, 25 » to 30 w in diameter. The clear, transparent shell has a thickness somewhat over half a micron. The embryo proper measures 15 w. Lying close against the inner surface of the shell is an ill-defined layer of flocculent substance, within which are three or four, possibly more, nuclei 2.5 » to 3.5 » in diameter, each containing a large nucleolus. A six-hooked embryo (fig. 14), from the specimen furnished by Dr. Stiles, possessed an outer shell about 45 » in diameter, and a somewhat thicker (2m) inner envelope 22 @ X 27 mw in diameter, with hooks 12 mw in length. A very thin membrane, which usually lies so closely applied to the outer en- velope as not to be distinguished, sometimes, under the influence of changes in osmotic pressure, ‘draws away from the latter and thus becomes apparent, as shown in fig. 14. INTERNAL ANATOMyY.—The rostellum (figs. 12, 16, ros) is bounded by a layer of muscles (fig. 12, rm), some of which are longitudinal ON HYMENOLEPIS CARIOCA AND H. MEGALOPS 161 and some circular. In the parenchyma of the rostellum, just be- neath the cuticula lining its cavity, is a dense layer of cells (fig. 12, rc), resembling somewhat the subcuticular cells elsewhere, but much more thickly crowded, and with more prominent nuclei. Nervous System.—The nervous system shows an arrangement re- sembling in general that of Taenia crassicollis and T. perfoliata as described by Cohn (98). My observations agree less closely with those of Tower (00). The main lateral nerves (fig. 12, Jn) in the scolex bend toward one another and are joined just behind the ros- tellum by a transverse commissure (co), which consists of two par- allel strands of nerve fibers (fig. 17) as in 7. perfoliata. The two halves of the commissure are connected by fibers running diag- onally from one to the other, and between them lie numerous ganglion cells. Surrounding each sucker near its posterior pole is a circular ring or zone of nerve fibers, lying in the same horizontal plane as the transverse cerebral commissure, and joined to it at its point of union with the lateral nerve. Fibers from these nerve zones run in various directions over the surface of the suckers, and at intervals tufts are given off which extend radially toward the sur- face of the head (fig. 17). In the regions where the zones approach one another they are connected by interlacing fibers. From the transverse cerebral commissure near the median line, a dorsal and a ventral pair of parallel nerves (fig. 17, dd, vv) come off at right angles and extend almost horizontally dorsad and ventrad respec- tively to the nerve zones, joining them in the region where they ap- proach nearest the median line. Fibers which cross at right angles between the two strands of the commissure establish a continuity between these dorsal and ventral nerves. This cerebral complex corresponds roughly to that of Taenia crassicollis as given by Cohn (98). While the cerebral commissure connecting the main lateral nerves in T. crassicollis is long, in T. megalops it is short and moreover double as in Taenia perfoliata. The two pairs of nerves (dd, vv) mentioned above form a dorso- ventral commissure composed of two parallel right and left halves. The dorso-ventral commissure in T. crassicollis is single in the region where it crosses the transverse commissure, but separates fork-like dorsally and ventrally. The inner quadrants of the nerve zones of the suckers correspond to the commissures in T. crassi- collis, which connect the main lateral nerves with the forks of the 162 B. H. RANSOM dorso-ventral commissure. The remaining portions of the zones together correspond to the “polygonal commissure.” Nerves ap- parently homologous to the accessory lateral nerves (fig. 12, na) join the cerebral complex some distance laterad from the main lat- eral nerves, about 90° measured along the circumference of the nerve zones. In T. crassicollis, on the other hand, the main lateral nerves and the accessory nerves of each side have a common point of origin. The dorsal and ventral median nerves join the complex (fig. 17, mm) near the ends of the dorso-ventral commissure, as in T. crassicollis. The median and accessory lateral nerves are con- nected behind the complex by numerous commissures with one an- other and with the main lateral nerves. In T. megalops, while these eight nerves are prominent through some distance just behind the cerebral complex, they seem to disappear entirely in the posterior part of the scolex, and if they are present in the proglottides are too small and insignificant to be traced. From the point at which each lateral nerve joins the cerebral commissure, two nerves (fig. 12, rv), the “apical branches’? (Cohn) of the lateral nerves, extend forward on each side to join the rostellar ring (figs. 12, 16, mr) right and left. From each of these apical branches a nerve (rm) comes off to enter the rostellum. The apical branches of the median nerves join the ros- tellar ring dorsally and ventrally (fig. 16). From the point of union of each of these eight nerves with the ring, a branch of nerve fibers passes outward over the surface of the adjacent sucker. A number of nerves extend forward from the same points. The lateral nerves are joined in three regions (fig. 19, ag, mg, pg) in each proglottis by dorsal and ventral commissures, as in T. perfoliata. The posterior commissures (fig. 18, pdc, puc) are the most prominent and lie between the two longitudinal muscle layers, while the other two pairs lie just within the inner layer (fig. 20, adc, avc). In connection with each commissural ring of the pro- glottis in T. perfoliata, all three of which lie inside the inner longi- tudinal muscle layer, Cohn describes a second ring which lies be- tween the muscle layers and joined with the former by radial fibers passing between the muscle bundles. It is this outer ring which forms the posterior commissures in T. megalops. An inner poste- rior ring, if present, is scarcely developed. Fibers faintly indicating such a ring are sometimes seen (fig. 18, 1). Musculature.—From the suckers powerful muscles of the inner ON HYMENOLEPIS CARIOCA AND H. MEGALOPS 163 longitudinal layer extend backwards grouped at first mostly in eight bundles (fig. 12, im). This muscle layer, which is very heavy and prominent in the anterior segments (fig. 18, im), becomes progres- sively thinner posteriad by the gradual reduction in size of the bundles (fig. 20, im). The outer layer (fig. 18, 20, om), consist- ing of a great number of small bundles, completely envelopes the inner layer and is distinctly separated from it. The bundles of this layer are of about the same size throughout the strobila, and hence in the posterior segments (fig. 20) are nearly or quite as large as the inner bundles. The origin of the outer layer is similar to that of H. carioca. There are no diagonal fibers. Dorso-ventral fibers are most prominent and numerous in the posterior portion of the proglottis. Transverse fibers are not present except a few in the pos- terior region (fig. 18, tm), and a few in the anterior region (fig. 15, tm), some of which have a special connection with the cirrus pouch. Excretory System.—The dorsal canals lie some distance directly dorsal to the ventral canals (figs. 15, 18, 20). The former measure 15 » to 20 pw in diameter in the anterior segments, and 9 p to 15 p in the posterior segments; the ventral canals in the most anterior segments have a diameter the same as that of the dorsal canals, while posteriorly they measure from 20 mw to 35 w. A transverse canal connects the ventral canals in the posterior part of each seg- ment. There is a basket-like plexus of small canals (figs. 15, 19, xpl) in connection with each ventral canal in the posterior half of the proglottis. In the scolex the canals of both sides turn inward and approach each other as they pass forward. The dorsal canal (fig. 11, dc) soon turns outward again and describes a loop around the posterior surface of the sucker, returning back toward the ventral sur- face close behind the cerebral commissure. From the region of the dorsal vessel where it passes behind the commissure two small canals (figs. 11, 17, er), one dorsal and one ventral, extend forward to enter the rostellum near its base, each joining in the anterior portion of the latter a corresponding canal from the dorsal vessel of the other side. There are accordingly two closed loops of the excre- tory system extending forward into the rostellum, as in Hymeno- lepts carioca and H. diminuta, but quite different from them in that they form connectives right to left, instead of dorsal to ventral. 164 B. H. RANSOM The dorsal canal continues toward the ventral surface, passes around the posterior face of the ventral sucker, and after completing the circuit curves outward and forward to pass anteriad between the dorsal and ventral sucker (fig. 17, dc). The ventral canal (fig. 11, vc) forms first a loop behind the dorsal sucker, posterior to and smaller than the loop of the dorsal vessel, then, returning toward the ventral surface, forms a second loop behind the ventral sucker also smaller than and posterior to the corresponding loop of the dorsal vessel. The returning limb of the dorsal loop of the ventral vessel crosses the outgoing loop on its anterior side. It is connected with the latter near the region of crossing by two or three small vessels, and continues ventrad as the outgoing limb of the ventral loop. The recurrent limb of the ventral loop turns outward like the corresponding limb of the dorsal vessel, passes behind the latter, and extends between the suckers toward the lateral surface of the scolex, finally turning anteriad to run forward in the same frontal plane as the dorsal vessel, between it and the surface of the head (figs. 11, 16, 17, vc). During this portion of the course of the two canals they are connected by numerous smaller vessels which pass at intervals from the dorsal canal forward and outward to the ventral vessel. The dorsal canal of each side finally unites with the ventral canal to form a loop right and left of the rostellum. The loops lie in nearly the same transverse plane. In two specimens examined the dorsal limb of the right-hand loop came from the ventral vessel, of the left-hand loop from the dorsal vessel (fig. 16, broken lines). Reproductive Organs.—The three testes (figs. 15, 19, 20, ¢) are located posteriorly and toward the dorsal surface in the median field, ordinarily two to the left and one to the right of the median line. They are at their maximum size in segments 160 to 170, where they measure 90 » to 115 » through their largest diameter. The vasa efferentia are wide, short tubes uniting to form the vas deferens (fig. 15) which extends forward near the median line. Before entering the cirrus pouch it passes beyond the base of the latter and then folds back to form a small loop in which are commonly found one or two vesicular enlargements (fig. 15, vs), representative of the prominent seminal vesicle of Hymenolepis carioca or diminuta. The cirrus pouch is a large, elongated, cylindrical structure at- taining a maximum size of about 300 p» in length by 60 w in diameter. It extends from the median line near the anterior end of the pro- ON HYMENOLEPIS CARIOCA AND H. MEGALOPS 165 glottis, passing with the vagina above the excretory canals and lateral nerve, backward and outward to the genital pore. A very thin layer of longitudinal muscle fibers forms its outer boundary. In mature segments the vas deferens is expanded within the cirrus pouch to a large seminal vesicle (fig. 15, vs’) which almost com- pletely fills its posterior portion. The vas deferens continues distally trom the seminal vesicle as a tube 15 w to 20 » in diameter, upon the outer surface of which is a layer of small nuclei. After a short distance it bends back toward the base of the pouch, gradually be- coming smaller, and, reduced finally to a diameter of 3 p» to 4 pn, opens into the proximal end of the cirrus. The cirrus (fig. 15, cz) is a long, powerful organ, slightly coiled, equal to or greater than the cirrus pouch in length, and measures 15 » to 20 w in diameter. It possesses a thick wall, smooth outwardly but lined inside with stout bristles, best developed toward its outer end, where they are 6 p to to w in length. The arrangement of the organs within the cirrus pouch is similar to that of Drepanidotaenia lanceolata (Wolffhigel ooa). Most of the fibers of the transverse muscle system in this region extend between the left side of the proglottis and the base of the cirrus pouch (fig. 15, br). The remaining few (tm) continue across to the other side of the segment. Within the pouch is found a further system of fibers (fig. 15, cr) extending between the prox- imal end of the cirrus and the base of the pouch. Some of them are apparently continuations of the other fibers (br) just men- tioned. These two aggregations of fibers seem to serve the functions of retractors of the cirrus pouch and cirrus, respectively. The genital cloaca is from 30 w to 50 deep. Into it open the cirrus and the vagina, the former posterior and dorsal to the latter. Although the opening of the vagina (fig. 19, vg) into the cloaca is very narrow, the lumen of its distal portion is wide and capacious, a condition which becomes more pronounced as the segments grow older, and the vagina begins to function as a sperm receptacle. It is covered by a prominent layer of nuclei. The widening of the vagina is accompanied by the appearance of ridges or rugae of the cuticula, the largest of which project 4 m to 5 w into the cavity. They are very thin and close together, and run around the vagina in a circular direction, so that in a longitudinal section they appear as a thick coat of cilia. It is likely that in many cases in which cilia have been described lining the cuticula-covered inner surface of the 11 166 B. H. RANSOM vagina, it is a question, not of cilia, but of projections similar to these ridges. At the base of the cirrus pouch the vagina, becoming narrowed to a diameter of 10 mto 15 uw, turns backward beneath the former, and passes posteriad to a point beneath the yolk gland (fig. 15, Vg). This organ is a compact, ovoid body with a maximum size of 90 p, lying towards the dorsal surface, beneath the points of union of the vasa efferentia of the testes. The shell gland (fig. 15, sg) is about 50 w in diameter, and lies immediately beneath the yolk gland. The ovary (fig. 20, ov) is a simple sac-like structure, slightly elongated transversely, with a depression behind giving it a some- what lenticular shape. From segment 158 to 168 the ovary in- creases in size very rapidly. In the former it has a transverse diameter of but 60 p, in the latter it has attained a width of 210 up, a thickness of 100 p, and a length of 135 yw. It lies in the ventral half of the proglottis, entirely anterior to the posterior limits of the yolk and shell glands. The maximum size of the ovarian ovum is 12) pe The uterus develops in front of the yolk and shell glands, imme- diately dorsal to the ovary. In segment 160 it is represented by a little lenticular mass of cells 50 w in diameter, lying ventral to the base of the cirrus pouch, and connected with the shell gland by an almost straight cord of cells 60 # in length. In the segments imme- diately following, the anterior mass of cells has begun to arrange itself into a layer enclosing a central cavity; the uterine duct from the shell gland has grown longer, and become bent back and forth. The uterus continues to increase in size, and in segment 168 extends forward to the anterior border of the proglottis, a distance of 125 yp, and measures in the transverse diameter 180 ». The second follow- ing segment (170) has a well-defined layer of cells upon the outer surface of the uterus, and this layer (fig. 19) becomes progressively thicker and more prominent toward the posterior end of the strobila. As the eggs pass out of the ovary it shrinks rapidly, and in segment 174, where all the eggs have entered the uterus, has entirely disap- peared. The uterus expands forwards and outwards toward the surface, and more slowly backwards, as a simple sac without infold- ings, differing thus from Hymenolepis carioca and H. diminuta. In the last dozen segments of the worm, partial ruptures occur ON HYMENOLEPIS CARIOCA AND H. MEGALOPS 167 in the continuity of the strobila (fig. 19), caused perhaps by the sudden contraction of the segments when the worm was killed. In these segments the anterior end of the uterus is more or less ex- posed. Whether such action takes place by natural muscular con- tractions is uncertain, but there is certainly a tendency in this di- rection. Consequent upon the separation of a ripe segment from the strobila, however brought about, a large area of the surface of the uterus is laid bare, and in this area a thin membrane is all that sep- arates the embryos from the outer world, facts which may play a part in their dispersal. With regard to the systematic position of T. megalops, while it differs considerably in a number of points from the type of Hymeno- lepis, it is scarcely possible at the present time to define its affinities more accurately than to give it, for the sake of its three testes and unilateral genital pores, the place of an aberrant species in the above genus. ON THE GENUS HYMENOLEPIS The genus Hymenolepis in its present status is composed of a considerable number of species all characterized by the possession of three testes and unilateral genital pores. Cohn (99, 99a, 99b, 00a) divides the genus thus characterized into two subgenera upon the basis of the number of hooks. Upon a priori grounds alone such a division seems exceedingly artificial, and indeed Wolffhigel (00, 00a) has demonstrated by two or three cases that it is false and misleading by reason of the non-correspondence of the superficial characters taken by Cohn to more important internal relations. There is still further evidence in this regard. H. carioca is clearly so similar to H. diminuta as to belong in the same genus. Since the rostellum is unarmed it would also, follow- ing Cohn’s classification, fall in the same subgenus Hymenolepis s. str. Specimens in my possession of Hymenolepis sp.? from one of the gulls, Larus Franklinit(?) possess a freely extensible ros- tellum armed with 10 hooks of the form typical of the now disused genus Dicranotaenia Raill. Upon the basis of Cohn’s classification we should refer it at once to the subgenus Drepanidotaenia. The form possesses three testes and in the details of cirrus pouch, the two seminal vesicles, seminal receptacle, ovary, yolk gland, and anlage of uterus shows striking similarities to H. carioca. There are the same 168 B. H. RANSOM number of inner muscle bundles and a definite system of diagonal muscles as in the latter. These two forms are very closely allied, and are much more similar to one another than to H. diminuta. The structural resemblance is certainly too great to allow of their separa- tion into two different subgenera, if one of the subgenera is also to contain H. diminuta. To arrange them according to Cohn’s system it would be necessary either to neglect their internal anatomy or to sup- pose that H. carioca in young stages possesses 8-10 hooks which are afterwards lost. That the latter circumstance occurs, however, seems improbable, otherwise we should not expect to find a specimen, as already noted, retaining hooks on the suckers, and at the same time with no trace of armature upon the rostellum. The foregoing facts, I believe, furnish a very good demonstration of the untenability of Cohn’s division of the genus. A division may be necessary, but it must be made upon a different basis than that attempted by Cohn. Apart from this question it is not even certain that he has wisely modified the definition of the genus in restricting the number of testes to three. In Blanchard’s description the number of testes is given as very few, most often three, which allows a desirable de- gree of latitude to this character. The form described by Jacobi (98) as Taenia inflata Rud. pos- sesses but two testes, and hooks which, while of the same form, are only a third as large as those of the type specimens examined by Krabbe. A tape-worm which Cohn (98a) considers the true T. inflata Rud., agreeing in all respects with Krabbe’s description, pos- sesses three testes, and he is accordingly of the opinion that Jacobi has described another species. The latter is possibly to be identified with Taenia spiculigera Giebel 1866. A similar form with two testes, from Fulica americana, has hooks 37 p» in length, somewhat larger than those of Jacobi’s specimens, in this respect agreeing with T. spiculigera more closely than the latter. The suckers of my speci- mens are armed. In anatomical details this form agrees with Jacobi’s description. It has, however, a seminal vesicle outside the cirrus pouch, besides the one within described by Jacobi. Except for the lack of agreement in the number of testes, Taenia spiculigera(?) is very like H. carioca and but for this difference would undoubtedly fall in the same genus. That this difference is not so important as may seem is shown by the fact that the number of testes in segments of H. diminuta, normally three, sometimes varies in either direction ON HYMENOLEPIS CARIOCA AND H. MEGALOPS 169 to two or four, as Grassi (88) has shown. I am able to confirm Grassi’s observations, in part, as I have sections of H. diminuta showing four testes in some segments. It is therefore questionable whether such a difference is sufficient to separate generically two forms otherwise similar, whose lack of agreement in this regard may be explained as the result of a slight variation, which, although oc- curring but occasionally in one, has become permanent and normal in the other. According to this view Taenia spiculigera(?) should be brought under Hymenolepis. BIBLIOGRAPHY BRAUN, M. 94-00. Vermes. Bronn’s Klassen und Ordnungen des Thierreichs. Bd. IV, Abt. 1b. Cestodes. CouN, L. 98a. Untersuchungen iiber das centrale Nervensystem der Cestoden. Zool. Jahrb., Anat., XII., pp. 89-160, pls. VI-IX, 9 text figs. 99. Zur Systematik der Vogeltaenien. Cent. f. Bakt. u. Parasitenk., Bd. XXV, No. 12, pp. 415-22. 99a Zur Systematik der Vogeltaenien, II. Cent. f. Bakt. u. Parasitenk., Bd. XXVI, pp. 222-27. 99b. Zur Systematik der Vogeltaenien, III. Zool. Anz., Bd. XXII, pp. 405-8. 00. Zur Systematik der Vogeltaenien, IV. Cent. f. Bakt. u. Parasitenk., Bd. XXVII, pp. 325-28. ° CREPLIN, FR. 25. Observationes de Entozois. P. I. Gryphisw. 86 pp., 1 pl. (Brief synopsis in Braun 94-00.) 29. Novae observationes de entozois. Berolini. 134 pp., 2 pl. (Brief synopsis in Braun 94-00.) DIESING, C. M. 50. Systema Helminthum. I. Vindobonae. 680 pp. 64. Revision der Cephalocotyleen. Abth. Cyclocotyleen. Sitzbr. k. Akad. Wien, XLIX, I Abth., pp. 357-430. DUJARDIN, F. 45. Histoire naturelle des Helminthes, ou vers intestinaux. Paris. 654 pp., pls. I-XII. 170 B. H. RANSOM FUHRMANY, O. 95. Beitrag zur Kenntnis der Vogeltaenien. Revue Suisse de Zool., T. III. pp. 433-58, pl. XIV. GIEBEL, C. 66. Die im zoologischen Museum der Universitat Halle aufgestellten Ein- geweidewurmer nebst Beobachtungen uber dieselben. Zeitschr. f. d. ges. Naturwiss., Bd. XXVIII, Thl. Il, pp. 253-78. GRASSI, B. 88. Zaenia flavopunctata Weinl., Taenia leptocephala Crepl., Taenia diminuta Rud. Atti R. Accad. d. sc. Torino, Vol. XXIII, pp. 492- 501, 1 pl. JACOBI, A. 98. Uber den Bau der Zaenia inflata Rud. Zool. Jahrb., Syst., Bd. XII, pp. 95-104, 1 pl. LUHE, M. 94. Zur Morphologie des Taenien-scolex. Inaug. Diss. Konigsbergi. Pr. 133 pgs., 12 figs. 96. Zur Kenntnis der Musculature des Taenien-korpers. Zool. Anz., Bd. XIX, pp. 260-64, 4 figs. MAGALHAES, P. S. DE. 98. Notes d’helminthologie brésilienne 8. Deux nouveaux Ténias de la Poule domestique. Arch. de Parasit. I, pp. 442-49, 6 figs. MINGAZZINI, P. 99. Osservazioni generali sul modo di adesione dei Cestodi alla parete in- testinale. Atti. Accad. Lincei (5), Vol. 81, pp. 597-603, 6 figs. Also Arch, Ital. Biol., XXXII, pp. 340-50. Abstract Zool. Jahresbericht. STILES, CH. WARDELL. 96. Report upon the Present Knowledge of the Tapeworms of Poultry. Bull. No. 12, Bur. An. Ind., U. S. Dept. Agric., Washington, D. C., pp. 1-79, pls. I-XXI. TOWER, W. L. 00. The Nervous System in the Cestode Monzezia expansa. Zool. Jahrb., Anat., Bd. XIII, pp. 359-84, pls. 21-26. WOLFFHUGEL, K. 00. Beitrag zur Kenntnis der Vogelhelininthen. Inaug. Diss. Univ. Basel. pp. 204, pls. I-VII. 00a. Drepanidotaenia lanceolata (Bloch). Cent. f. Bakt. u. Parasitenk., Bd. XXVIII, pp. 49-56, 6 figs. ZSCHOKKE, F. 88. Recherches sur la structure anatomique et histologique des Cestodes. Mem. de 1’ Institut national. Genevois. T. XVII. pp. 396, pls. I-IX. ON HYMENOLEPIS CARIOCA AND H. MEGALOPS 171 EXPLANATION OF PLATES Figures outlined with camera except as stated otherwise. ABBREVIATIONS adc Dorsal anterior nerve commissure. ag Anterior ganglion of lateral nerve: an Anterior nerve from _ cerebral ganglion. avc Ventralanteriornerve commissure. ér Retractor of cirrus pouch. ci Cirrus. cg Cerebral ganglion. cv Transverse cerebral commissure. cp Cirrus pouch. cr Retractor of cirrus. dc Dorsal excretory canal. dd Dorsal halves of dorso-ventral commissure. dm Diagonal muscles. er Excretory loops of rostellum. fga Anlage of female glands. gc Genital cloaca. 7m Inner longitudinal muscles. Zu Main lateral nerve. mg Middle ganglion of lateral nerve. mp Muscle bands. Myoblasts. ma Accessory lateral nerve. In fig. 16, point of union with cerebral complex. mn Median nerve. In fig. 16, point of union with cerebral complex. nr Rostellar nerve ring. Outer longitudinal muscles. ov Ovary. pce Prostate(?) cells of vas deferens. pdc Dorsal posterior nerve commis- sure. pg Posterior ganglion of lateral nerve pr Protractors of cirrus pouch. pve Ventral posterior nerve commis- sure. rc Subcuticular cells of rostellum. rm Muscle layer of rostellum. yn Nerve to rostellum. vos Rostellum. rp Cavity of rostellum. run Apical branch from main lateral nerve to rostellar ring. Sg Shell gland. sm Limiting membrane of cirrus pouch. sy Seminal receptacle. ~ )-festis. tm Transverse muscles. ut Uterus. vv Ventral halves of dorso-ventral commissure. ve Ventral excretory canal. vad Vas deferens. ve Vas efferens. vg Vagina. vgs Vaginal sphincter. vs Seminal vesicle. In fig. 4, vesicle of cirrus pouch. vs’ Seminal vesicle of cirrus pouch. xpl Plexus of excretory vessels. x Dorsal posterior inner nerve com- missure. yg Yolk glands. 172 B. H. RANSOM Plate XXIII Flymenolepis carioca Fig. 1. Ventral view of segments. Toto preparation. 245. Fig. 2. Dorsal view of very young segments. ><245. Fig, 3. Hooks from suckers. Free-handdrawing. ><2600. Fig. 4. Cirrus pouch and seminal receptacle. Transverse section. ><1100. Fig. 5. Hook from onchosphere. Free-hand drawing. ><2000. Fig. 6. Onchosphere with membranes. From specimen in formalin. ><535 Fig. 7. Transverse section through anterior region of proglottis. ><600. Plate XXIV HT, carioca Fig. 8. Scolex. Toto preparation. 185. Fig. 9. Ventral view of posterior end of strobila. Toto preparation. ><50. Fig. 10. Frontal section of scolex. 180. FT, megalops Fig. 11. Excretory system in scolex. View from anterior. Reconstruc- tions from transverse sections through scolex behind the rostellum. ><55. Fig. 12. Frontal section of scolex. ><55. Fig. 13. Young embryo. 600. Fig. 14. Six-hooked embryo. 600. Plate XXV HT. megalops Fig. 15. Dorsal view of mature segment (No. 172). Reconstruction from sections. 135. Fig. 16. Transverse section through the anterior region of scolex. The broken lines are anterior to the plane of section. 45. Fig. 17. Transverse section of scolex at level of cerebral commissure. 45. Fig. 18. Transverse section through the posterior region of one of the first 35 segments. >< 120. Fig. 19. Frontal section through a mature segment (No. 180). ><120. Fig. 20. Transverse section through the anterior region of a mature seg- ment (No. 168). >120. PLATE XXIII ‘ - PLATE XXIV ap MS Gs ea aes Oe =e a BHR. del, ty 7 2 hy PLATE XXV BHR.del. STUDIES ON THE GENUS CITTOTAENIA By RUFUS ASHLEY LYMAN WITH TWO PLATES The parasites studied were collected from rabbits in Nebraska and Kansas. A careful record was kept to give some idea of the degree of infection, and, although the number of rabh’’s examined is not large enough for entirely satisfactory results, , . some interesting data have been obtained. For this purpose 87 rabbits were exam- ined; 47 were found to harbor parasites, 3, containing cestodes, 31 cysticerci, and 3 nematodes. The rabbits were of two different species, the common cotton-tail, Lepus sylvaticus, and the common jack, L. melanotis. Of the 60 rabbits killed in eastern Nebraska, 55 were cotton-tails and 5 jacks. Of the 55 cotton-tails, 38 were infected, but none of the jacks were. On the other hand, of the 27 killed in southwestern Nebraska and adjacent parts of Kansas, 12 were jacks and 15 cotton-tails. Of these 3 cotton-tails were infected, while the number of jacks infected was 7. The cestodes taken from these rabbits represent four forms: Cittotaenia variabilis, C. variabilis angusta from L. sylvaticus, and C. pectinata and a single-pored form from L. melanotis. Several specimens of the single-pored form were obtained, but were so poorly preserved that final determination of the species must be left until more material is obtained. The cestode, however, occupies a unique position, as the following facts show. Each proglottis always contains only one set of female glands. The genital pores are unilateral and always on the right margin of the strobila. In this it resembles the genus Anoplocephala, but differs from it as regards the position of the female glands and the distri- bution of the testes. In Anoplocephala the testes are located in the aporose, the ovary in the pore side of the proglottis (Stiles 96) ; but in the present form the testes are distributed throughout the 174 RUFUS ASHLEY LYMAN proglottis, and the ovary is located in the median line. This re- sembles somewhat the conditions found in Bertia americana Stiles 1896, but the testes extend the whole length of the proglottis as well as to the excretory canals on either side (fig. 9), and the female glands are always located in the median line in succeeding segments, and not right and left of the median line, as in B. americana and B. americana leporis. So far as has been ascertained, the degree of infection by cestodes remains about the same throughout the year, at least for the species Cittotaenta variabilis. This form has been collected at all seasons and seems to be about as frequent in the winter months as in the summer. Riehm (81) states that it is said that C. pectinata occurs in the European hares only in the fall and the first half of the winter, but the adult parasite was found in Rawlins county, .Kansas, as early as March and throughout all the spring and summer months. The number of worms found in a single rabbit seems to vary with different species. The largest number of C. variabilis found in a single rabbit was 5. They often occur singly, but generally in pairs. Twelve individuals of C. variabilis angusta were found in one rabbit. C. pectinata seems to occur in greater numbers. Goeze mentioned the fact that 20 or 30 are found in a single host. In the present case as many as 54 were taken from one rabbit. THE GENUS CITTOTAENIA This genus was first proposed by Riehm in 1881. In it and in Dipylidium R. Leuckart he placed the double-pored forms of rabbit cestodes, while the single-pored forms he placed in Taenia. How- ever, in the same year, he rejected Cittotaenia as a genus and placed the only species in Dipylidium. In 1891 R. Blanchard proposed a new genus, Moniesia, based upon the number of genital pores. In this genus he included the double-pored forms of cattle, sheep, and allied animals and also T. festiva R. 1819 from Macropus giganteus, T. marmotae Frélich 1802 from Arctomys marmota, and T. pecti- nata Goeze 1782, D. Leuckarti Riehm 1881, and D. latissimum Riehm 1881 from rabbits. Blanchard placed the single-pored forms in Anoplocephala. In 1893 Railliet proposed the new genus Cteno- taema, with T. marmotae as type, and placed here Ct. Goezet (=Riehm’s C. latissima), Ct. Leuckarti, and Ct. pectinata. The establishment of this genus was an error on the part of Railliet, who STUDIES ON THE GENUS CITTOTAENIA 175 evidently overlooked Riehm’s first paper. The same mistake was made by Stiles in 1895, and by Stiles and Hassall in 1896, who ac- cepted Railliet’s genus Ctenotaenia. Because of the many differ- ences between the double-pored cestodes of rodents and ruminants, Stiles in 1893 separated the double-pored forms of rodents, which had been placed in Moniezia by Blanchard, from that genus, but did not attempt to classify them. In 1896 he adopted Riehm’s for- merly discarded genus, Cittotaenia, for the double-pored forms, giv- ing it priority, of course, to Railliet’s genus Ctenotaenia. Stiles dis- tributed the single-pored forms among the genera Anoplocephala E. Blanchard 1848, Andrya Railliet 1893, and Bertia R. Blanchard 1891. Cittotaenia pectinata (Goeze 1872 partim, Riehm 1881) Stiles and Hassall 1896. The history of this parasite is thoroughly reviewed by Stiles (1896). The species has been reported from Germany and France, but up to the present instance it has not been reported in America, and Stiles (1896) considers his C. variabilis the American repre- sentative of C. pectinata. The parasites assigned to this species were collected in Buffalo county, Nebraska, and Rawlins county, Kansas, from Lepus mela- notis, and are certainly not identical with C. variabilis Stiles and Hassall (96), but resemble very closely the form described by Riehm as Dipylidium pectinatum, and by Stiles and Hassall! (96) as C. pectinata, differing only in some minor particulars, to which attention will be called in the description cf the various organs. External Characteristics——The adult strobilae do not vary greatly in size. In 54 specimens of various ages taken from a single rabbit the parasites range from 7 mm. to 71 mm. in length. The smallest is 7 mm. long and possesses 40 proglottides. The strobila is leaf-like in form, a characteristic which presents itself in all of the young forms and persists more or less perfectly as long as all the proglottides remain intact. The general shape gives one the impres- sion of a large liver fluke, being widest near the center of the body and gradually tapering toward both ends. In the larger specimens the anterior portion is comparatively narrow, becoming lanceolate. The posterior proglottides always become narrower and longer, often reaching 1.5-2 mm. in length. The strobila attains its greatest width, 176 RUFUS ASHLEY LYMAN 9-10 mm., about 15-20 mm. back of the head, and then remains com- paratively uniform in breadth until near the posterior end. Follow- ing is a table showing the length, the breadth in different regions, and the number of proglottides present in five typical strobilae. All measurements are given in millimeters. WIDTH OF TOTAL WIDTH 2 | WIDTH 10 | WIDTH IN NO. OF LENGTH OF| MM. BACK | MM. BACK | wipesT | 2OSTERIOR | procrot- STROBILA |OF SCOLEX |OF SCOLEX PLACE PROGLOTTIS TIDES 44 3 6 8.5 2.5 140 Every proglottis é present. 52 3.5 6 9.5 2 139 Every proglottis present. 68 3.5 5.5 7 4 | 125. | Posterior pro- glottis gone. 27 3.D th 8 4 110 Posterior pro- glottis gone. 71 4.5 7 9 8 140 Several gone. The proglottides are always broader than long. When the terminal proglottis is present, it has the form of a bulb applied closely to the preceding proglottis. At its extreme posterior part is a notice- able depression, the opening of the excretory canals. The posterior border of each proglottis is smooth and overlaps the following pro- glottis but slightly. The genital pores, two in each proglottis, are situated about the middle of the segment margin. They are never prominent, and usually can be distinguished only by the aid of a lens, and often only in section. The cirrus has never been found protruding from the pore. The head is dome-shaped (fig. 6) and its measurements are much larger than those given by either Stiles or Blanchard for C. pectinata. Stiles in his diagnosis gives 0.25 mm. for the diameter and 0.125 mm. for the thickness of the head, and says that the suckers are small, but gives no measurements. Blanchard (1891) gives meas- urements as follows: head, 315 jm to 340 » broad; suckers 142 p long by 135 » broad; opening 80 uw long by 53 » broad. In the present form the diameter of the head varies from 0.41 to 0.45 mm. ; the thickness from 0.28 to 0.31 mm. While the head is much larger, the suckers are small, being about 0.12 mm. long by 0.088 mm. broad. The cavity of the sucker measures from 64 to 74 » long by 28 to 34 » broad. In section the suckers are seen to open nearly STUDIES ON THE GENUS CITTOTAENIA 177 straight forward or at a very small angle outward. In all cases, from the smallest to the largest strobila, the neck is absent, segmen- tation beginning immediately at the base of the head. MuscuLature.—There are two dorsal and two ventral plates of longitudinal fibers (fig. 4,1 m). The outer plates are much the larger. The fibers tend to run in bundles, and single fibers often run through 10 to 40 proglottides. Certain fibers running in adjacent bundles branch off and form anastomoses with other bundles, and so a dense network of fibers is formed. The inner longitudinal muscle plate is not as highly developed as the outer. Near the lat- eral margins of the strobila, the plates of the dorsal and ventral sides approach each other and finally meet. The inner plate always remains separate from the outer, but its fibers become much less distinct at the margin of the proglottis. Directly within the inner longitudinal plates there are two transverse plates of fibers which run entirely across the proglottis and enclose the male and female reproductive glands (fig. 4, ¢ m). A third system is present as sagittal fibers (fig. 4, s m). They run dorso-ventrally, forming a dense network in the parenchyma between the dorsal and ventral transverse plates. They branch greatly, and these branches form a network around the testicles and the various female glands. Many of them pierce the transverse and the longitudinal plates. Excretory SystemM.—There are four longitudinal canals, two dorsal and two ventral. These persist throughout the anterior, the middle, and most of the posterior portion of the strobila. In the extreme posterior portion they become so branched that it is often impossible to distinguish dorsal canals from ventral. This is quite different from the description given by Stiles, who, in his diagnosis (96), says that the dorsal canal was not observed. Riehm (81) remarks that the dorsal canal becomes obliterated some distance from the head. In the present form the dorsal and the ventral canals are nearly the same in size. The diameter of the ventral canal varies somewhat, while the dorsal remains more nearly con- stant. The diameter of the ventral canal varies from 34 » to 48 p the dorsal from 34 # to 40. The dorsal canal (fig. 4, d c) lies dorsal and slightly median of the ventral, about 0.75 mm. from the margin of the proglottis. In the head the dorsal canals unite, like- wise the ventral. The dorsal canals pass dorsal to the nerve ganglion, while the ventral canals run ventral to them, and the two 178 RUFUS ASHLEY LYMAN canals formed by the union of the four unite between the suckers, forming a single canal, which ends as a blind tube near the anterior point of the head (fig. 2, b ec). A loop is formed here by the canals around the ganglia, and the blind sac is an anterior projec- tion from the point where the ducts unite. The ventral canals are connected by a transverse canal in the posterior portion of each proglottis. The transverse canals of adjacent proglottides are again connected by secondary longitudinal canals, which often branch and © give the parenchyma the appearance of being divided into little islands (fig. 7). This network of secondary longitudinal canals is more prominent in the anterior than in the posterior portion of the strobila. When the posterior proglottis is attached, the excretory canals come together, forming a more or less irregular reservoir (fig. 3, 7), which opens at the excretory pore through many canals. Nervous SystemM.—Two ganglia are located in the head just back of the suckers. They lie between the arms of the loop formed by the dorsal and the ventral canals (fig. 2). Each ganglion is about 53 » long by 40 w wide. They are connected by transverse commissures. Small nerves are given off anteriad to the suckers. The main nerve trunks leave the ganglia at the sides, run toward the margins of the head, and turn backward. There are three dis- tinct longitudinal nerves present. Two run ventral to the genital ducts and one dorsal. The main trunk lies just outside of the ventral longitudinal canal (fig. 4, m m1). It runs in about the middle of the strobila in the dorso-ventral direction except, where it passes beneath the genital canals, it dips slightly ventrally (fig. 5,7 n). Its average diameter in the anterior region of the stro- bila is 60 w. In the posterior portion of each proglottis there is a ganglionic enlargement from which nerve fibers are given off. Some run in the direction of the genital pore, while others pass toward the median portion of the proglottis, but their course can not be traced. No transverse proglottidal commissures have been found. The dorsal and the ventral trunks are about the same size, averag- ing 13 m in thickness. The ventral lies directly below the main trunk, and the dorsal directly above, but dorsal to the genital ducts. Both secondary trunks are connected with the primary by commis- sures in the posterior portion of each proglottis (fig. 5,¢ 0). From this section the relative position of nerve trunks, commissures, and EEE ee STUDIES ON THE GENUS CITTOTAENIA 179 genital ducts can be seen. The posterior commissures are constant. Just behind the vagina and cirrus pouch are the other commissures connecting the main trunk with the secondary trunks, but these are not as well developed as the posterior commissures and are not as constant. MALE REpRODUCTIVE OrGANS.-—The anlagen of all the genital ducts appear in the proglottides immediately behind the head. The anlagen of the testes appear first about 6 mm. or 7 mm. from the anterior extremity of the head. They develop repidly and do not atrophy in the posterior region of the strobila, the capsules nearly always remaining in perfect form. Even in the posterior proglottis they are found grouped around the excretory pore and between the canals leading to the pore (fig. 3,¢). There are from 100 to 125 in each proglottis, arranged in a quadrangle confined to the dorsal portion of the proglottis and posterior to the uterus. They extend posterior from the ovaries to the longitudinal canals (fig. 1). This is a characteristic of much importance, since Stiles quotes it as one of the important differences between the European form, C. pec- tinata, and the American representative of the same species, C. variabilis, in which the testes extend to, but not beyond, the ovaries. They are nearly spherical in form, averaging 70 m in diameter, but often attaining a thickness of 92 », Running from each testis is a small duct, which joins the transverse tubule. There are two of these tubules on each side of the proglottis. One receives the ducts from the testes lying in the middle portion of the proglottis, while the other receives those from the testes lying back of the ovary and those lying externally toward the longitudinal canals. These tubules join posterior to the ovary and form the vas deferens. The vas deferens runs dorsal to the ovary in an outward oblique direction toward the anterior end of the proglottis. Its usual length is 0.75 mm. to I mm. In the younger stages it is not twisted, but later it becomes greatly convoluted (fig. 4, v d), forming loops in the dorso-ventral direction only. The vesicula seminalis is quite variable. Often there are one or more small swellings in the vas deferens just before reaching the cirrus pouch, but, when present, they are very much smaller than the figures of Riehm would indicate. There is usually just inside the cirrus pouch an enlargement which, in some cases, nearly fills the pouch and is filled with a dense mass of sperm. 180 RUFUS ASHLEY LYMAN The nozzle-shaped cirrus pouch is very large, extending some distance mediad from the longitudinal canals (fig. 1, ¢ p). Its length varies from 0.925 to 1.075 mm. and its width from 65 p to 85 ». The pouch is muscular, consisting of two sets of muscle fibers (fig. 8). The outer layer consists of longitudinal fibers; the inner, which is also the thicker, consists of circular fibers. The cirrus wall averages from 5 w to 24min thickness. It contains both circular and longitudinal fibers arranged similarly to those of the cirrus pouch. Although the cirrus pouch lies dorsal to the vagina, it does not open dorsally to the latter, but the vagina so twists around that its opening is posterior to that of the cirrus and in the same frontal plane (fig. 1). The sheath of the cirrus is composed of spongy tissue (fig. 8, s ¢). Riehm (81) in one of his figures indicates both the cirrus and the vagina as opening externally into a comparatively shallow cloaca. In the present form the cloaca is a narrow duct opening upon the top of a small cone which is situ- ated at the bottom of the genital pit (fig. 1). The genital pit or pore is situated near the middle of the margin and is very indis- tinct. There are no projecting lips and, in many cases, no depres- sion is present. The largest pits measure 56 w in diameter and 32 yp deep. FEMALE REPRODUCTIVE OrGANS.—These organs appear earlier than those of the male. The anlagen of the glands are found in the proglottides immediately behind the head, but the uterus does not ap- pear until somewhat later. Back of the middle of the strobila all of the female organs, except the uterus, begin to atrophy, and in the posterior proglottides only remnants of the vagina remain. The ovary, shell gland, and vitellarium lie about 1.6 mm. from the mar- gin of the proglottis, nearer the ventral side, and nearly fill the pro- glottis in antero-posterior diameter. Each proglottis contains two sets of female glands. The vagina lies ventral to the cirrus pouch, the distal end becom- ing posterior and opening behind the cirrus. At its distal end, the lumen is very small and the cuticular lining very thick. As it runs mediad the lumen widens and the walls become thinner until it reaches a point just within the longitudinal excretory canals, when its walls become very thin, and it expands into the external recepta- culum seminis, the first receptaculum seminis of Riehm (fig. I, e rs). This thick-walled portion of the vagina is surrounded by a STUDIES ON THE GENUS CITTOTAENIA 181 thin layer of circular muscle fibers, and outside of the fibers is a single layer of cells which stain deeply. These cells, near the ter- mination of the vagina, have the appearance of pavement epithelium, but farther mediad they become columnar (figs. I and 8, d s c). The cuticular lining of this portion of the vagina is covered by cilia (fig. 8c 1). The combined length of the first portion of the vagina and the external receptaculum is about I mm., or equal to the length of the cirrus pouch, and they both vary in the same ratio. The widest portion of the external receptaculum is 40 w. After reaching the external receptaculum the character of the vagina changes greatly. The deeply staining cells entirely disappear, the walls be- come very thin, and no muscle fibers are present. After leaving the external receptaculum the vagina is smaller in diameter as it runs mediad until it reaches the internal receptaculum, the second of Riehm (fig. 1,2 r s). The narrow portion is about 0.625 mm. long. The internal receptaculum does not differ from the external in structure, but is larger, averaging 208 » long and 80 p» in diameter. The internal and the external receptacula and the portion of the vagina between them is always full of sperm. The shell gland surrounds the oviduct just at the entrance of the vitelline duct. In cross-section it is seen to consist of spindle-shaped cells surrounding the oviduct. Its diameter varies from go to 100 p. The vitellarium lies posterior and dorsal to all other female glands (fig. 1,v ¢). It is more or less bean-shaped and divided into lobes or pouches by infoldings of its walls. Its length is about 200 4, its width, 90 ». The vitelline duct is about 25 » long and leaves the gland from its concave side. The contents of the gland are finely granular and stain deeply. The ovary consists of numerous pouches which extend anteriad laterad, and ventrad, but is located dorsally and posteriorly by the sides of the vitellarium. In these individuals the ovary is not bi- lobed as stated by Riehm, and the shape of the pouches is different from that indicated by his figures. They are Indian-club shaped, averaging about 112 » long and being widest, 16 », at their distal ends. Each pouch is connected with the common reservoir (fig. 1, ro) by a narrow neck only large enough for one ovum to pass. The common reservoir is an oval structure about 57 » wide by 95 pw long, into which all the pouches empty, and from which the oviduct 182 RUFUS ASHLEY LYMAN leads. The whole ovary, together with these pouches, is about 0.475 mim. long by 0.15 mm. wide. The oviduct (fig. 1, 0 v) is very simple. It runs posteriad from the reservoir of the ovary toward the yolk-gland. Its course is slightly convoluted. Beyond the opening of the vazina and the vitelline duct into the oviduct, the latter passes through the shell gland, at the same time turning forward, becomes the ootype, and enters the uterus just below the ovary. Its diameter is about I0 p. There is never more than one uterus in a single proglottis. In the early stages, it is a simple, transverse, rod-like organ which runs dorsal to all other organs and extends laterally beyond the excretory canals. In older proglottides the walls of the uterus become slightly digitate by the growth of shallow proximal and distal pouches, which are always simple. The ripe proglottis is almost completely filled by the uterus. The ova are more or less irregular or polyhedral in form, prob- ably due to pressure or shrinkage, since in the younger proglottides they are spherical. They measure 50 to 58 » in diameter, and the bulb of the pyriform body is 16 to 21 w thick. The horns are long and usually filamentous. Ciitotaenta variabilis (Stiles 1895) Stiles and Hassall 1896. Some additional points have been worked out in the structure of this species. This form shows variations in almost every possible direction. The strobila many attain a length of 17 or 18 cm. The head, though quite constant in form, varies greatly in size. It is spherical in general form, but slightly flattened on top and at the sides. In the present forms it varies from 0.462 to 0.872 mm. in diameter. The suckers are not only variable in size but also in shape. They are more commonly spherical, about 0.2 mm. in diameter, with a cavity 0.094 mm. across. Sometimes they are slightly elliptical, measuring 0.282 mm. long by 0.106 mm. broad, but this elliptical form may be due to contraction. MuscuLATURE.—The muscular system is even more highly de- veloped that in C. pectinata. The muscle fibers are larger, run in larger bundles, and lie closer together. A subcuticular layer of fibers, consisting of fibers running in both the longitudinal and the transverse directions, is especially well developed. STUDIES ON THE GENUS CITTOTAENIA 183 ExcrETORY SystEM.—This system is simple, consisting of four longitudinal trunks which persist throughout the strobila. The ventral canal is thin-walled and much the larger. In cross-section it is seldom circular, but is usually longer in the dorso-ventral direc- tion (fig. 1ov c). Its dorso-ventral diameter averages about 0.217 mm.; the transverse, 0.095 mm. The dorsal canal is smaller, and, in a true transverse section, circular, or nearly so, in outline. It lies dorso-median of the ventral canal, and its average diameter is 60. The character of its walls is very different from the ventral canal, since it has a thick cuticular lining around which is a thick layer of muscle fibers (fig. 10, d c). All canals unite in the head between the suckers, forming a loop. There is, however, no an- terior projecting canal from their junction, as in C. pectinata. The ventral canals are connected in the posterior portion of each pro- glottis by a transverse canal, but no secondary longitudinal canals connecting the transverse canals are found. Nervous SystEM.—The material used for the study of the nervous system in this species was preserved in vom Rath’s killing solution and some additional points were obtained, but such struc- tures as the proglottidal nerve rings, which Tower (97) describes for Moniezia expansa and M. planissima, could not be traced. The ganglia occupy the same position behind the suckers as in C. pectt- nata, but the commissures connecting them can be seen more dis- tinctly. Anteriorly there are given off nerves which run around the suckers and toward the anterior point of the head. A cross-sec- tion of the head (fig. 11) shows the position of the ganglia with reference to the dorsal and the ventral longitudinal canals. They lie between the canals which pass forward to unite between the suckers. In this species, also, three longitudinal nerve trunks are present, two running ventral and one dorsal to the genital ducts. But, instead of the three lying in the same sagittal plane as in C. pectinata, the two secondary trunks lie outside of the main trunk toward the margin of the proglottis (fig. 10, mn, dn, vn). No com- missures could be found connecting the main and the secondary trunks as in C. pectinata, but the inability to trace them may be due to the thickness of the sections and the density of the stain. A ganglion is found on the main trunk in the posterior part of each proglottis, from which nerve fibers run toward the genital pore and are lost in the tissue around the pore. Others are given off from 184 RUFUS ASHLEY LYMAN the inner side, but their terminations can not be found. Smaller branches are given off throughout the course of the nerve (fig. 12). Mace REPRODUCTIVE OrGANS.—The testes number from 75 to 100 in each proglottis and are confined to the posterior side of the uterus and to the dorsal portion of the strobila. They extend to, but never beyond, the ovary; they are oval in form, measuring about 80 » by 100 w. Each one is surrounded by a thick layer of muscle fibers, the probable function of which is to force the sperm out of the testis into the duct, as in C. pectinata. The vas deferens is convoluted in the dorso-ventral direction only (fig. 10, vd), but it has no peculiar characteristic. There are no enlargements found in its course, the small vesicula seminalis being situated just inside the cirrus pouch. It is rarely more than 12 to 20 » in diameter. The cirrus pouch (fig. 13) may reach a length of 0.4 mm., but it is usually much less. Its diameter is more nearly uniform through- out its length than in C. pectinata, often being 50 ». Its muscular walls are highly developed, there being an outer longitudinal layer of fibers and an inner circular, the combined thickness of both often reaching 16 p. The cirrus is from 5 » to 8p in diameter. It opens dorsally to the vagina (fig. 10). In this species the cloaca is surrounded by a sphincter muscle, the wall band of which is 13 pw thick (fig. 13, sp m). Its probable function is to contract in self-fertilization and to force the male products to pass up the vagina. The genital pore is situated in the middle of the lateral margin. Although it is not prominent, it is easily detected, since the margin of the proglottis is depressed near the pore. Usually there is a little pit (fig. 13, g p), or pocket, opening by a narrow neck, but the lips may be so closely applied to the inner surface of the pocket that the cavity is obliterated. The pit measures 13 » deep by 21 p wide. FEMALE REPRODUCTIVE OrGANS.—The vagina opens into the cloaca ventral to the cirrus (fig. 10, v), and lies ventral to the cirrus pouch throughout its length. It is about 10 pw thick, and there is no enlargement corresponding to the external receptaculum of C. pec- tinata, the organ remaining narrow and of about the same caliber until it swells into the receptaculum near the ovary. The first 0.7 or 0.8 mm. of the vagina is surrounded by deeply staining cells as STUDIES ON THE GENUS CITTOTAENIA 185 in C. pectinata. The receptaculum seminis varies in length; its average is about 65 p. The ovary, shell gland, and vitellarium are situated about 1.3 mm. from the lateral margin and resemble the corresponding organs in C. pectinata, but are usually longer in transverse diameter and nar- rower in an antero-posterior direction. The yolk gland is elliptical in form, 0.266 mm. long (1. e., across the proglottis) and 0.088 mm. wide. The ovary is 0.69 mm. long, in the same direction, and 0.1 mm. wide. The ovarian tubules are much heavier than in C. pectinata, being from 37 to 45 » wide and from 75 to 100 » long, and are connected with the common reservoir by thick necks (fig. 10,0). The shell gland occupies a position corresponding to that in C. pectinata, and all the female glands are crowded closely together. The uterus may be single or double in the same strobila. In some cases, all, or nearly all, are single; in others, all, or nearly all, are double. The uterus appears as a single transverse tube which lies dorsal and extends beyond the excretory canals. In the older stages it becomes slightly and irregularly digitate, and finally en- tirely fills the proglottis. If two uteri are present, their blind ends become closely applied in the median line and can be distinguished only in section. The ova average about 58 » in diameter and the bulb of the pyri- form body, 16p. The horns are usually about 1.5 times as long as the pyriform body is thick, and are usually straight, do not cross, and are not filamentous. Cittotaema variabilis angusta. - This variety is only about 2 mm. broad, but often reaches a width of 3mm. The internal anatomy differs from that of C. variabilis principally in size, all the organs being proportionally smaller. There are a few peculiarities, however, both in the shape and position of certain organs which seem to be quite constant. 1. All the organs are situated in the posterior portion of the pro- glottis, leaving the anterior portion completely bare (fig. 22). 2. The genital pores are situated close to the posterior margin of the proglottis (fig. 22, g p). 3. The ovaries, in proportion to the width of the strobila, are situated far from the lateral margins, the distance from the lateral 186 RUFUS ASHLEY LYMAN margin to the ovary being almost as great as the distance between the ovaries. Consequently the testes are more closely crowded to- gether (fig. 22). 4. The ovary has a very characteristic appearance. The diameter of each pouch is nearly the same throughout its length. 5. The number of uteri in each proglottis within the same stro- bila is more variable than in the type of the species. PROGLOTTIDAL VARIATIONS.—A study of the variations in the pro- glottides shows some interesting facts in the various forms. In all the specimens of C. pectinata, the external appearance of the pro- glottides is perfectly normal. No irregularly developed or abnormal proglottides appeared, and no cases of intercalation, such as Blanch- ard’s (1891) figures show, are found. Concerning the number of sets of reproductive glands, a very peculiar facts presents itself. Each of the last two proglottides of the strobila always contain but one set of reproductive glands, and also the genital ducts open on opposite sides of the strobila. In C. variabilis the external appearance of the strobila is quite different. Instead of all the proglottides being even and regular, as in C. pectinata, there are many irregular and abnormal ones and many cases of intercalation. More than half of the strobilae possess abnormal proglottides and often twenty abnormalities occur in a single strobila. The simplest form of intercalation is seen in fig. 14. Here the proglottis, a, is represented only by a small lip on one side of the strobila and it possesses no internal organs. Fig, 15 shows a more typical case of intercalation. The proglottis, a, is the inter- calated one, but from the external appearance of the strobila in this region ac would seem to be the normal one, and b the abnormal. The internal anatomy, however, the arrangement of the testes, and the perfectly formed transverse excretory canal show that bc is the normal proglottis. The proglottides a and be are nevertheless closely connected. Although the transverse canal of a is lost in the paren- chyma, the testes of @ are continuous with those of bc, and the uterus of a dips backward into the parenchyma and between the uteri of bc. It should be noted that all of the uteri in this strobila are double. Fig. 16 shows a more peculiar case. The proglottides ab and ef are evidently normal. Now the question is whether cd is one proglottis with but one set of sexual glands, or is ¢ an inter- calated proglottis with one set of glands, and d another intercalated - ~ J se a ee STUDIES ON THE GENUS CITTOTAENIA 187 proglottis with no sexual glands whatever. The latter view seems more probable since the transverse excretory canal of ¢c in about the middle of the strobila turns forward and unites with that of ab, while d possesses neither sexual organs nor transverse excretory canal. Fig. 17 is a subcuticular section of the same proglottides, which shows that the margin of proglottis d is connected with ab and that of c with ef. Fig. 18 is a section through three proglot- tides. Ab represents one proglottis with only one set of reproduc- tive glands, but it should be noticed that the proglottis is not normally developed on the side a. Fig. 19 shows a section through three proglottides of another strobila. The section was cut obliquely, so the three fully developed sets of organs of the left side are not shown. On the right side of section ab the reproductive glands are pushed mediad so that they lie just inside of the glands of the preceding and of the succeeding proglottides. The ovary, shell gland, and yolk gland are perfectly developed, and a portion of the receptaculum seminis is also present, but all genital ducts, as well as the genital pore, is absent. Testes are present but no vas deferens. Just how the eggs developing in this ovary are to become fertilized is a question. This proglottis seems to represent a stage intermedi- ate between that represented in fig. 18 and a normal proglottis with two fully developed sets of female glands. Fig. 20 shows a proglot- tis ab with all the organs developed, but the margin on the right side is not developed, and the genital pore opens between the pre- ceding and the succeeding proglottides. Fig. 21 shows a case, the most highly modified of all in some respects. The genital ducts on the right side of the proglottis cd, instead of opening on the normally developed margin d, turn forward and open with the ducts of the proglottis ab, so that four genital ducts open at the one genital pore. Unfortunately, the proglottis was so old that it was impossible to determine the exact relations of the four genital ducts. The variety angusta also shows many abnormalities and intercala- tions similar to those of C. variabilis and occasionally normal pro- glottides with but a single pore. The fact that proglottides are found with but a single set of re- productive organs is important in that it shows that proglottides with single and with double sets of generative organs may occur in the same strobila. But why in the case of C. pectinata the single sets should always occur only in the last two proglottides and why 188 RUFUS ASHLEY LYMAN the genital pores should always be on alternating sides of the stro- bila are questions yet to be explained. It hardly seems possible that these two proglottides can represent only one proglottis when the fact is considered that there are two sets of testes (fig. 3, t), two uteri (fig. 3, w), and that the transverse excretory canal (fig. 3, tr) in next to the last proglottis is perfectly developed. i —— STUDIES ON THE GENUS CITTOTAENIA 189 WORKS CITED BLANCHARD, R. 91. Notices, etc. 7. Cestodes du groupe des Anoplocephalinae. 8. Sur les Moniezia des rongeurs, Mem. Soc. Zool. France, IV, p. 443-66, figs. 21-35. RAILLIET, A. 93. Traité de zoologie médicale et agricole. 1fasc. II Ed. RIEHM, G. 81. Studien an Cestoden, Zeitschr. f. d. ges. Naturwiss., 3, VI, pp. 345- 610, Taf. V-VI; also Inaugural Dissertation. STILES, C. W. 95. Notes on Parasites. 36. A double-pored Cestode with occasional single pores, Cent. f. Bakt. u. Paras., 1 Abth. XVII, pp. 18-14, 457-59, 1 fig.; also in The Veterinary Magazine, II, 1895, 222-25, 1 fig. 96. A Revision of the adult Tapeworms of Hares and Rabbits, Proc. U. S. Nat. Museum, XIX, pp. 145-235, pls. V-XXV. STILEs, C. W., AND HASSALL, A. 96. Notes on Parasites, 47; On the priority of Cittotaenia Riehmm 1881, over Ctenotaenia Railliet 1893, The Veterinary Magazine, III, p. 407. TOWER, W. L. 96. On the Nervous System of Cestcdes, Zool. Anzeiger, XIX, pp. 323-27. 190 RUFUS ASHLEY LYMAN EXPLANATION OF PLATES Plates XX VI, XXVII EXPLANATION OF ABBREVIATIONS USED bec Blind pouchofexcretorycanals. ov Oviduct. c Cirrus. Excretory reservoir. ct Cilia. ro Reservoir of ovary. cm Circular muscle fibers. Sg Shell gland. co Nerve commissures. séc Secondary longitudinal canals cp Cirrus peuch. of the parenchyma. dc _ Dorsal excretory canal. sm Sagittal muscle fibers. adn _ Dorsal nerve. st Spongy tissue of cirrus sac. asc Deeply staining cells. sp m Sphincter muscle. ec Excretory canal. t Testes. ep Excretory pore. ‘mm Transverse muscle fibers. ers Ext. receptaculum seminis. iy Tranverse excretory canals. g£ Ganglion. u Uterus. gp Genital pit or pore. v Vagina. z Island of parenchyma. vc Wentral excretory canal. 7rs Int. receptaculum seminis. vd _ Vas deferens. Zm Uongitudinal muscle fibers. v2 Ventral nerve. mn Main nerve. vs Vesiculo seminalis. 7) Ovary. vé¢ Vitellarium. ot Ootype. vtd Duct of vitellarium. EXPLANATION OF FIGURES All figures are made from camera drawings. Figs. 1-8. Cittotaenia pectinata. Fig. 1. Dorsal view of male and female reproductive organs. X79. Fig. 2. Frontal sections of scolex through ganglia and excretory canals. x93. Fig. 3. Frontal sections through posterior proglottis. 12. Fig. 4. Transverse section of proglottis. 32. Fig. 5. Sagittal section through nerve trunks. 62, Fig. 6. Head. 60. Fig. 7. Secondary longitudinal canals of one proglottis. > 68. Fig. 8. Transverse section of cirrus pouch and vagina. 293. Fig. 9. Single-pored form. Frontal section of one proglottis. ><30. Figs. 10-21. C. variabilis. Fig. 10. Transverse section througn proglottis. 30. Fig. 11. Transverse section through head. 43. Fig. 12. Frontal section through strobila showing main nervetrunk. ><32. Fig. 13. Frontal section through cirrus pouch and sphincter muscle. 93. Figs. 14, 15, 16, 17, 18, 19, 20, and 21. Abnormal proglottides. v7. Fig. 22. C. variabilis angusta. Y¥rontal section of proglottis. 57. PLATE XXVI t be aan eet Uf ie om hen 4oae tm : -im PLATE XXVIt eee EE AS pera SOME POINTS IN THE STRUCTURE OF THE ACANTHOCEPHALA By H. W. GRAYBILL WITH ONE PLATE In the singular group of parasitic worms known as the Acantho- cephala, the large nuclei of the subcuticula and lemnisci are most interesting and unique structures. They lie in a mass undivided by cell walls and in the adult worm are usually present in great num- bers. Even in the same individual they may present an astonishing number of sizes and forms. In size they vary from moderately small structures to enormously extended bodies, as in the lemnisci of E. clavula Duj. (Hamann 91). Their form may be either spher- ical, ovoid, or amoeboid. These nuclei divide directly, and probably have to some degree the ability to move (Hamann g1, 95). In the living parasite they have been observed to push out and withdraw pseudopodia-like processes (Kaiser 93, p. 29). They are not only found in the substance of the lemnisci and subcuticula but are fre- quently seen lying in the lacunae of the same. In the subcuticula and lemnisci of an echinorhynchus collected from the rock bass (Ambloplites rupestris) and black bass (Micropterus dolomieu) of the Great Lakes are found nuclei of such extraordinary size and form that an extended description of them seems called for. It will suffice here to say that the subcuticula of this worm is of the ordinary type, being bounded externally by a thin cuticula and internally by a delicate basement membrane (membrana limitans). The radical and the peripheral fiber systems are well developed. Near the membrana limitans is located the usual set of canals, con- sisting of two large lateral vessels extending nearly the whole length of the animal and a meshwork of lacunae connecting the two both dorsally and ventrally. The nuclei referred to above lie approxi- 192 H. W. GRAYBILL mately in the middle of the subcuticula immediately beneath the in- nermost layer of peripheral fibers. A stained mount of the body wall spread out shows, under low magnification, along the middle of both the dorsal and the ventral sides, a deeply stained, well-formed, dendritic mass (fig. 12), which appears at first sight to extend continuously almost from one end of the body to the other. On closer examination, especially under a higher power, it is found that each of the supposed single masses is divided into several parts which follow each other more or less closely. A glance shows that these several long branching bodies (nuclei) are independent of the lacunae which lie in a different level and form a complete network between the lateral canals. Each nucleus consists of a more or less sinuous main trunk, which lies about midway between the two lateral canals. It is not uniform in width, but is successively contracted and expanded, having a ‘maximum width of 14 mw. On either side are given off, about at right angles, a large number of branches which may be simple or variously branched. The branches usually come from the nodes of the main trunk. They are at times moderately uniform in width, while again they may be contracted at one or more places to a minute fiber. In fact, under low magnification, many small round- ish or irregularly shaped portions of nuclear matter appear to be disconnected from the surrounding branches. However, under a high power it is found, in all but a very few cases, that a drawn- out thread of nuclear matter definitely connects each of these to a near-lying branch. Some of the few truly isolated portions had cer- tainly separated from the main mass just previous to or at the time of killing, as a drawn-out point of nuclear matter from the isolated part pointed directly toward a similar projection on a neighboring branch. The width of these branching nuclei is such that they occupy ap- proximately the middle half of the spaces between the lateral canals. In some cases, however, the branches approach quite close to these canals, especially when the ends of two successive nuclei pass each other somewhat (fig. I2). The various branches of the nuclei are so arranged with respect to the net-work of lacunae that they occupy as much as possible median positions in the islands of hypodermis between the lacunae. The number of dendritic nuclei in the body varies from about ten Se ee eee i? a — ee eee ee STRUCTURE OF THE ACANTHOCEPHALA 193 to twelve. In length they vary in good sized worms from 2 to 4.6 mm. They stop short some distance from the anterior and pos- terior ends of the body, so that in sections from these regions no nuclear matter is encountered. In each lemniscus is found a single large nucleus. The lemnisci are very long, thread-like bodies. Each is divided into two moder- ately distinct regions ; a proximal, broader, flat region, 450 to 900 p long, and a distal cylindrical region many times longer than the former. The large nucleus is located in the proximal portion of each organ. It is, roughly speaking, of a branching type and in length occupies nearly the whole of this region of the lemniscus. In sections its outline is usually dentate or irreguiar (fig. 10). Each nucleus is formed of a longitudinal portion of very ununi- form width with a maximum of 56. Attached along the sides of this are a few short branches which are in some cases expanded at their ends into broad masses. The lacunae of both regions of the lemnisci are frequently filled with a granular coagulum which at most stains very lightly. It is interesting to note here that Hamann (g1, p. 146) found only a single nucleus in each lemniscus of E. clavula Duj. The measurements given by him for this nucleus are, length 1.6 mm., breadth 0.15 mm. However, the nuclei of the subcuticula were of the usual type and showed well the various division stages. (The writer is very greatly indebted to Dr. Otto Hamann of Berlin for slides of this, as well as of various other species, which he so kindly forwarded at the request of Dr. H. B. Ward.) The nuclear substance of the above described branching bodies stains very uniformly. Only under the highest power is it found to be compactly granular in structure. It contains a moderate number of vacuoles of various sizes which usually are bounded by definite circular outlines (figs. 3, 5, 7, 10). Chromatic bodies or nucleoli are present in the nuclear substance in countless numbers. In form they vary greatly, being spherical, ovoid, irregular, or long and thread-like. Those of the first three types predominate. They vary in size from the minutest granules to structures 22 w long. Those filamentous in form, though only about 1.5 to 3 » in width, may, in exceptional instances, reach a length of 84 ». They are not always uniform in width throughout. Frequently they are very weakly expanded and contracted at inter- 194 H. W. GRAYBILL vals along their whole length. In some cases this is so pronounced as to give a distinctly moniliform appearance to the chromatic body. In fig. 9 are represented some of the irregular forms of the above chromatic bodies. The remaining figures will readily give an idea of other forms. They stain very deeply. They may contain one or several small roundish transparent specks which are probably vac- uoles. Even after the stain has almost entirely washed out of the nuclear matter no structure is to be detected in these chromatic bodies which are still quite deeply stained. At times in the nuclei of the lemnisci the roundish chromatic bodies occur in groups, and in some cases they lie in contact, form- ing rather compact masses. In general the larger chromatic bodies and frequently also the smaller ones are surrounded by unstained halos which have usually a distinct boundary, the same as the vacuoles spoken of above (figs. 1-8). These halos are not so frequent in the nuclei of the lemnisci. Hamann (91) describes and figures similar halos in a nucleus from a lemniscus of E. clavula Duj. Some of these chromatic masses are suspended in the halos by minute chromatic strands which radiate from them, connecting with the nuclear substance (figs. I and 2). In cross-section the different branches and the trunks of the branching nuclei are circular, oval, or oblong. And since in sagittal sections of the body, especially just to one or the other side of the main trunks of these nuclei, most of the branches are shown in cross-section, an appearance of the subcuticula is presented not un- like that found in echinorhynchi where the nuclei are comparatively small in size and large in numbers. The extensive branching brings portions of these dendritic nuclei into most of the regions of sub- cuticula, and also gives to them an enormous surface area for the carrying on of metabolic processes. In another species of echinorhynchus (host—Mo.zostoma sp.) which is related to E. proteus West, among the many spherical, ovoid, and amoeboid nuclei of the subcuticula were also found a few very large ones which were dendritic in form. The longest one measured had a length of 630 » Some of the medium sized ones of this type possessed as many as seven good sized branches. A comparison of the branching nuclei in the first form consid- STRUCTURE OF THE ACANTHOCEPHALA 195 ered above, with the nuclei in Neorhynchus clavaeceps (Zed.), and N. agtlts (Rud.) is interesting. According to Hamann these two forms represent sexually mature larvae. Each possesses six to ten large (0.2 mm.) ovoid nuclei in the subcuticula of the body and two in each lemniscus. In the form studied the number of nuclei is also small, there being ten to twelve in the body and one in each lem- niscus. As suggested above, the branching is probably both for the purpose of increasing the superficial area and bringing portions of the nuclei into all regions of the subcuticula. The total amount of nuclear matter is immeasurably greater than that contained in the few ovoid nuclei of N. agilis and N. clavaeceps and is probably about the same as in cases where the subcuticula contains large numbers of small nuclei. In the development of the nuclei of the subcuticula in the echino- rhynchi as worked out by Hamann (91) and Kaiser (93) one may distinguish roughly three stages. In the first stage the nuclei are large, roundish bodies and few in number. In the second stage the number of nuclei is still small, but their form is amoeboid with pseudopodia-like processes extending out from the various sides. The third stage is characterized by a large number of small nuclei of various sizes and forms and is brought about by an extensive fragmentation of the amoeboid nuclei of the previous stage. How- ever, though this is the usual mode of development, Hamann has pointed out in the case of N. clavaeceps and N. agilis, forms already referred to, that in the adult the nuclei have remained at the first stage of development. In view of this development it therefore seems probable that the above described dendritic nuclei have arisen by an extensive growth of the few large amoeboid nuclei of the larva and that they occupy an intermediate position between the nuclei of the two forms last mentioned above and the nuclei of other forms where they are present in great numbers. EXPERIMENTS ON THE Lemnisct.—As stated above, the lemnisci of the echinorhynchus collected from the rock bass and the black bass are very long, thread-like organs. In some cases they reach posterior to the middle of the body. They are unattached except at their points of origin. No muscular layer is present. The single large canal of the posterior or distal portion of the lemniscus di- vides into two canals on entering the proximal region. These latter 196 H. W. GRAYBILL canals unite at the attached end of the organ and empty as one canal into the circular canal at the base of the neck. The latter canal con- nects with the lacunae of the neck and is separated from those of the body by a cuticular ring. Some experiments were made on living material to determine what effect the cutting off of these well-developed organs would have on the extrusion of the proboscis. The first experiments consisted in cutting the worm in two im- mediately posterior to the proboscis sheath, thus severing both lem- nisci. A turbid fluid issued from the cut ends of the latter. While in many cases, because of the great injury to the parasite, no motion of the proboscis took place, yet in some few cases the proboscis was completely extruded a number of times. In numerous second experiments a slit was made in the body wall a little posterior to the base of the neck. Through this open- ing the lemnisci were drawn out and cut off close to their points of origin. The injury to the worm was slight. In many cases it was found that immediately or after the lapse of some time the pro- boscis was completely extruded and withdrawn many times with a rapidity equal to similar movements in uninjured worms. Also, in observing the movements of the fluid in the lacunae of the neck and proboscis of uninjured worms during the extrusion of the latter, it was found that the streaming anteriad always com- menced after the beginning of the extrusion and never preceded it. Again, when extrusion ceased, the fluid of the lacunae for some time continued to stream anteriad and adjust itself as it would not have done if under pressure sufficient to have aided in the extrusion. From the above observations it seems plain that in this form at least the lemnisci take no part in the protrusion of the proboscis (Hamann 91, p. 150), and also that any pressure on the fluid of the body cavity is not necessary to the operation. DESCRIPTION OF THE SPECIES.—With the aid of material kindly sent by Dr. E. Linton to Dr. H. B. Ward, for purposes of com- parison, the writer was able to determine definitely the identity of the form concerned in this paper with that from Roccus americanus described by Linton (92, p. 528) as Echinorhynchus thecatus. As the original description is very brief, being based almost entirely on toto mounts of the parasite and on the measurement of only a few individuals, the following more extensive list of the important char- STRUCTURE OF THE ACANTHOCEPHALA 197 acters, including measurements of hooks and of a large number of individuals, will be of great systematic value. Living specimens flattened dorso-ventrally, smooth. Color white, light yellow, orange, or gray. Preserved specimens cylindrical, curved. Proboscis also curved when extruded. Transverse wrinkles only on anterior half of body. Maximum diameter of body in female toward anterior end, in male at or slightly anterior to middle of body. In both sexes gradual decrease in diameter to an- terior end where neck joins abruptly. Similar decrease posteriad in case of male. End moderately blunt. In female posterior end slightly expanded, very blunt. Neck increases in diameter to base. Proboscis usually broadest at middle. Anterior end rounded. Hooks, 24 to 31 transverse and 12 longitudinal rows, surrounded by prom- inent collars. Collars of hooks at the base of the proboscis fre- quently longer than hooks. Recurved base of hooks notched at end. Lemnisci long and thread-like. Proboscis sheath double-walled. Ganglion in midst of proboscis retractors, midway between anterior end of proboscis and base of sheath. Nuclei of circular muscle layer arranged in two lateral bands, one opposite each lateral canal. Testes contiguous, anterior one the longer. Cement glands in com- pact mass, eight in number. Embryos spindle-shaped. MEASUREMENTS WIDTH LENGTH ANTERIOR MAXIMUM POSTERIOR Body, female.| 11-26 mm. 0.51-0.89 mm.) 0.8-1.4mm. | 0.52-1 mm. male.. 7-12 mm. 0.39-0.69 mm.} 0.59-0.95 mm.| 0.37-0.75 mm. Weekes s)3 5c. 179-298 p- DOE D9 8) beat 2) sere aiame ane Sis ape 254-388 pe Proposeis....| 868-1164 6- | .0.05.25..2..6. 239-328 p. pesca ee Embryos .... SHSLOS) ene ilere see tare mete rataraes TBD 2i sr. 1. Wein te erases Hooxs.—In the following table in the case of those hooks of the ventral row with recurved bases, of the two possible measurements of the thorn and the recurved root the longer one is given. The hooks with recurved bases in the dorsal row are measured from corresponding points, just as if the anterior projection were not present. The measurements at the curve were made along the line of maximum width. Values are given in microns. 18 198 H. W. GRAYBILL LONGITUDINAL ROW ON VENTRAL SIDE OF LONGITUDINAL ROW ON DORSAL SIDE THE PROBOSCIS OF PROBOSCIS Transverse Length Length re- | Diameter Length Length re- row thorn curved root | at curve thorn curved root a | | | 2 72 65 29 65 44 “resale ehereene 4 712 58 29 66 Bl). “Sarees 6 76 69 29 72 AQ): illsxeteisusgietae 8 716 he 29 72 40)" newer 10 19 76 32 72 32) NA Soe e Length of Width at foto) base 12 76 76 31 Bay uh heres uae 25 14 2 62 27 (ERA kane tabs sl 22 16 69 48 22, (MMO) aa slarrs coc 20 18 65 27 20 TO eee 20 Length of Width at hook base 20 GBC tarts cietiteia 22 GP ei nerratette 14 22 GBre ths |Heememnct ar 18 GL Wit yee eee 13 5 FON MARA 14 48 | es 13 Hosts.—Micropterus dolomieu and Ambloplites rupestris; stom- ach, pyloric coeca, intestine, and body cavity. Amiéa calva in in- testine. Free or attached. The above work was done in the Zoological Laboratory of the University of Nebraska under the direction of Dr. Henry B. Waxa, to whom the writer is under great obligations for many valuadle suggestions and for the use of a very extensive collection. STRUCTURE OF THE ACANTHOCEPHALA I99 WORKS CITED HAMANN, OTTO. 91. Die Nemathelminthen. Monographie der Acanthocephalen (Echino- thynchen). Erster Teil, Jenaische Zeitschrift, XXV, 119 pp., 10 pls. 95. Monographie der Acanthocephalen (Echinorhynchen). Zweiter (Schluss) Teil. Hermann Costenoble, Jena, 1895. 40 pp., 4 pls. KAISER, J. E. 93. Die Acanthocephalen und ihre Entwickelung. Bibliotheca Zoologica VII, 303 pp., 10 double plates. LINTON, EDWIN. 92. Notes on the Entozoa of Marine Fishes, with Descriptions of New Spe- cies. Part III, Acanthocephala. U.S. Com. Fish and Fisheries, Part XVI (for 1888), pp. 523-38, 8 pls. 200 H. W. GRAYBILL EXPLANATION OF PLATE Plate XXVIII Drawings outlined with camera lucida. Figs. 1-8. Longitudinal sections of various portions of dendritic nuclei of hypodermis. > 600. Fig. 9. Variously shaped chromatic bodies found in the same. ><600. Fig. 10. Longitudinal section of a portion of nucleus in a lemniscus. 600. Fig. 11. Hooks of proboscis. ><540. a. Type found on lateral and dorsal (convex) surfaces of anterior half, 6 and e. Hooks from ventral (concave) surface, anterior portion. d and c. Hooks from posterior portion. Fig. 12. Microphotograph of flat mount of a portion of the dorsal half of the body wall. Limits of a single nucleus indicated by the arrows. Knds of two adjoining nuclei also shown. Lateral canals and net work of lacunae rep- resented by the light lines. PLATE XXVIII AX ERS LOZ Days Reds D's, sha : Soy eee ve if AN Py CSS far J ses RES THE NORTH AMERICAN SPECIES OF CURVIPES By ROBERT H. WOLCOTT WITH FIVE PLATES I. INTRODUCTION The members of the genus Curvipes are distinguished from those of all other genera of water-mites by their strength, activity, and ferocity, other species of water-mites, small insects, larvae, various species of crustacea, and, in fact, almost any aquatic form which they can master falling a victim to their rapacity, and many of these being superior in size to their captors. The species are readily identified by the possession of structural characters which, though not subject to wide variation, are easily sufficient to distinguish the different species one from the other. These are the possession of palpi the fourth segment of which bears two or more papillae, and which are terminated by three or four small claws; of epimera collected into four groups of two each; of a genital area immediately behind the last epimeron, which pre- sents a median area flanked by numerous acetabula set free in the body wall or imbedded in chitinous plates; of legs bearing swim- ming-hairs and ending in retractile claws; and, in the male, of modified segments of the two last pairs of legs—the sixth in the case of the third pair, the fourth of the other, this modification being of a characteristic type. The species agree very closely in structure, and it is well to give a general description of the characters of the genus, leaving to be in- cluded in the descriptions of the individual species only such details as are subject to specific variation and distinguish each particular form. The body is elliptical or oval in form, moderately compressed or highly arched, the anterior margin more or less emarginate be- tween the eyes, and in some forms with posterior lateral emargina- 202 ROBERT H. WOLCOTT tions. There is also frequently a dorsal ccnstriction behind the eyes. The surface is usually marked with fine lines, and in some forms there are evidences of the deposition of chitin in the sub- cutaneous tissue. In front of the eyes and below them are usually found two bristles varying in length and form and termed, from their position, “antenniform bristles.” The maxillary shield, seen from beneath and forming the floor of the so-called “capitulum,” is of the typical shield shape with a well- developed ancoral process extending backward between the anterior pair of epimera. The mandibles are flat and four-sided with the ventro-posterior angle more or less produced, while at the anterior end is articulated a claw, which is curved, triangular in section, and tapers to a point. On the inner surface of this claw is a patch of what usually appears like fine, parallel striae, but which fortunate views resolve into a comb of fine hairs (pl. X XIX, fig. 6), and along the lateral edge of the flexor or concave surface is a series of fine serrations. The palpi are usually compressed, especially the fourth segment, the flexor margin of which is provided with two or more papillae, together with a spur at the distal end. There is usually a small spine at the tip of the extensor margin of the first segment, and on the second three spines in a row on the inner surface and two on the outer, one in the middle and the other at the distal end. The third segment has two spines on the inner surface and one on the outer, while the last segment bears at the tip three or four small, more or less curved, claws. The second segment is the thickest, the third and first about equal in this regard, the fourth next, and the distal the narrowest, while in length the fourth usually is great- est, and then in order the second, fifth, third, and first. The epimera are always collected into four masses, the first and second and third and fourth of each side being in close apposition. In the male the space between the second and third is usually nar- row and that between the two posterior of opposite sides narrow or quite obliterated, while in the female the spaces separating the four masses are all of them relatively wide. The posterior margin of the last epimeron is produced caudad, forming a sharply pointed angle within which the margin of this epimeron is usually more or less concave, while laterad of it the margin runs almost straight te the articuiation of the fourth leg. THE NORTH AMERICAN SPECIES OF CURVIPES 203 The legs in the female exhibit an increasing length and also an in- creasing breadth from first to last, while the individual segments decrease gradually in thickness from the base outward. The last segment of the first three pairs of legs in this sex is more or less club-shaped and bears at the outer end two claws which are capable of being retracted into a cleft on the extensor side. These show a long pointed tip between which and the base is usually a second, blunt, more or less flattened and curved tip, and at the base a broad, flat expansion. Beneath this claw at the tip of the segment on the flexor side is often a short straight spine, while on the middle of either margin of the cleft that receives the claw is a pair of fine hairs, near the outer end of these margins another pair, and at the inner end of the cleft still another. The legs are provided with a double row of weak spines along the extensor surface and long, heavy spines on the flexor surface, which increase in number from the first to the fifth segment, and on the basal segments tend to be grouped at the tip, on the third in the middle also, and on the fourth and fifth scattered along the margin in a double row. There is also a greater or lesser number of long, slender hairs on the posterior side of the fourth and fifth, and sometimes the third segment of the anterior pairs of legs, while in the posterior one or two pairs these are gathered into a close oblique row at the tip, become of value in progression, and may be termed “‘swimming-hairs.” The sixth segment of the last pair of legs in this sex is usually long, slender, and straight and bears claws which are miniatures of those on the anterior legs. In the male the first two pairs of legs are similar to those in the female, and the third also, except that the fifth segment is usually elongated and devoid of long, scattered hairs or swimming-hairs, and the last segment is modified in a characteristic manner to serve as asperm-carrier. This modification consists in a shortening of the whole segment and more or less of a thickening of the tip, which is rather squarely truncate. The cleft which receives the claws is carried around on to the end of the segment, but retains the hairs along its margin described for the female. The claws are highly and variously modified in the different species, the two becoming frequently unlike. It is to be noted that all the elements present at the tip of the corresponding segment of the female are here present but in a modified form, no new structures being added. 204 ROBERT H. WOLCOTT In the case of the fourth leg of the male the four basal joints are usually thicker than the outer two, and the fourth is modified to form an organ for grasping the anterior legs of the female in copu- lation. This modification consists in a deep excavation on the pos- terior side, both proximad and distad of which the segment is often dilated. The anterior side is gently convex and along it are a few weak spines, while on the flexor surface there are the usual long, stout spines. Proximad of the excavation on the posterior surface are numerous stout, blunt, more or less curved, saber-shaped spines, varying in length, and a few longer, pointed spines. Within the ex- cavation are a few spines, while distad of it is a greater or less num- ber of short, stout, blunt ones, and a varying number of long, slender swimming-hairs. Owing to the fact that the last pair of legs is di- rected backward, the posterior surface, on which is the excavation, is next to the body, while the anterior surface is on the outside. The genital opening is close behind the last pair of epimera, and the chitinous plates about that of the male are frequently fused with the inner end of the last epimeron. In this sex the opening is gen- erally small and is either at the bottom of a depression which serves as a seminal receptacle or opens into a more or less well- developed seminal pouch. This median portion is usually flanked by chitinous plates varying in form and bearing a variable number of acetabula. The genital area of the female exhibits a long genitai cleft in the median line guarded by two flaps, which together form an elliptical area, and which in turn are flanked by chitinous plates bearing a corresponding number of acetabula. In a small number of species these acetabula, in one or both sexes, are imbedded free in the wall of the body. The anal opening is situated at a varying distance behind the genital area, and the chitinous ring surrounding it may be fused with the genital plates. The characters just given define the general appearance of any mite belonging to this genus. The sexes are easily separated by the presence of the characteristic leg segments of the male, and by the character of the genital area. In the separation of species the characters which seem to be of most value are as follows: The length, thickness, form, and propor- tions of the palpi, together with the characters of the papillae and spur on the fourth segment; the lengths cf the legs, relative THE NORTH AMERICAN SPECIES OF CURVIPES 205 and absolute; the form of the segments, and whether or not the end of any of them shows a flattened expansion of the anterior surface extending beyond the base of the next seg- ment; the character of the segments peculiar to the male; and the number and distribution of swimming-hairs. The form and details in structure of the genital area are also highly im- portant, and the position of the anal opening is of value. Size of body and other measurements compared with body proportions are of uncertain value, since when the eggs become developed the females are swollen to a great degree, and not only is the form of the body modified but the relative proportions of other parts to the dimensions of the body are altered and the distance between the epimera increased, while their relative size is lessened. In the beginning of this investigation the author undertook, from the comparison of a large number of specimens of C. Reighardi, to determine the limit of specific variation, and found specific characters to be remarkably constant, a fact which has been confirmed again and again in the comparison of individuals belonging to the other species studied. The genus is generally distributed over the world, a large number of species being known from various parts of Europe, several from eastern Africa and Madagascar, two from Ceylon, a few from Cen- tral America and Mexico, and two from Canada. There have been hitherto noted as occurring in North America the following species : Curvipes numulus (Stoll) (87: 47, pl. XI, fig. 2). Curvipes guatemalensis (Stoll) (87: 11, pl. X, fig. 2, pl. XI, he...) Curvipes alzatei (Dugés) (84: 345, pl. VILL, figs. 10-19). Curvipes fuscatus (Hermann) (04: 58, pl. V1, fig. 9). The two former were described by Stoll from Central America and the second later recorded by Koenike (95): 209) from Canada. C. alzatei was described from Mexico, while the last species in the list is a well-known European form which has been noted by Koenike (1. c.) as occurring in Canada. C. nuwmulus is quite different from any species described in this paper, having only one papilla on the fourth palpal segment and only two claws on the tip of the last segment of the palpus. The acetabula are imbedded free in the body wall in the case of the female, the male being unknown. Of C. alzatei the female and early stages were described; it is related, 206 ROBERT H. WOLCOTT according to Piersig (1901: 265), to P. conglobatus (Koch), the female of which also has the acetabula set free in the body wall and the fourth palpal segment of which is short and with prominent papillae, thus differing from C. coronis, described on a subsequent page of this paper. C. guatemalensis is closely related to C. rotun- dus (Kramer), but the palpus is thicker, the two papillae on the fourth segment are one behind the other, the distal segment has only two claws at the tip, the legs are shorter, and the hairs on the genital plates less numerous. C. fuscatus is in many ways similar to C. constrictus, also a species described in the following pages, but the genital area of the male of that species does not agree with that figured and described for C. fuscatus by Piersig (97 and 1901), ac- cording to whom the seminal pouch is broader than long and roughly triangular instead of elliptical. The female of C. fuscatus also has the plate bearing the acetabula lunate in form, while in C. constrictus it is solid, which serves to ally the latter with quite different species. The collection upon which the present paper is based has been gathered at different times during several years past and represents not only the collections of the writer but contributions from others to whom he is greatly indebted. The following localities are represented : MassacuuseEtTts.—Cranberry Lake, Woods Hole, August, Igoo. MicuiGAN.—Lake St. Clair, summer of 1893; pools near Ann Arbor, spring of 1894; Round, Pine, and other small lakes near Charlevoix, summer of 1894; Intermediate Lake, Ellsworth, August, 1894; High Island Harbor, northern Lake Michigan, August, 1894; Reed’s, Lamberton, and other small lakes near Grand Rapids, sum- mers of 1895, 1896, 1897, 1898, and 1899; Grand River, near Grand Rapids, summers of 1895 and 1898; Kawkawlin River and Saginaw Bay, August, 1895 (by J. B. Shearer) ; Les Chenaux Islands, north- ern Lake Huron, August, 1895 (by J. B. Shearer). Wisconsin.—Lake Winnebago, Oshkosh, August, 1897. ILLino1s.—Several localities near Havana (by the Biological Sta- tion, Illinois State Laboratory of Natural History). NEBRASKA.—Circle Lake, Decatur, June, 1899 (by Chas. For- dyce) ; pool in Monroe Canyon, Sioux county, May, 1899; South Bend, September, 1897 (by H. B. Ward). CoLorapo.—Pond near Wray, November, 1901. Missourt.—Ponds near Columbia, August, Igot. THE NORTH AMERICAN SPECIES OF CURVIPES 207 LovuistANA.—Pond in Audubon Park, New Orleans, fall of 1901 (by E. Foster) ; near Slidell, fall of 1901 (by E. Foster). To those whose names appear in the foregoing list as having as- sisted in the collecting of material the author is deeply grateful. The specimens have been preserved for the most part in one of the solutions recommended by Koenike, which leave the specimens in condition most suitable for dissection and she study of skeletal struc- tures; although precaution has been taken at the same time to pre- serve a greater or less number in picro-sulphuric, corrosive subli- mate, alcohol, or formol mixtures in order that the natural size and form might be preserved. In the preparation of slides the author has subjected them to treatment with potassic hydrate, removing the most of the contents from the body and separating and dissect- ing out the mouth parts, mounting all in balsam. In the case of single specimens of the species the legs and palpus have been re- moved from one side and mounted in balsam, care being taken that no pressure should be put upon the body in such a way as to alter its form or proportions, and leaving the appendages of the other side in the natural position. Through the courtesy of Dr. F. Koenike, of Bremen, Germany, the author has had specimens of the following European and other forms for comparison: C. conglobatus (Koch), C. wncatus Koenike, C. nodatus (Miller), C. guatemalensis (Stoll), C. rotundus (Kra- mer), C. variabilis (Koch), C. fuscatus (Hermann), and C. longi palpis (Krendowsky). The study of the material thus secured has resulted in the detection of fifteen species, of which thirteen are new. If we add to these the species heretofore recorded we get a total of nineteen for North America, as follows: 1. Curvipes coronis n. sp. 11. Curvipes medius n. sp. 2. Curvipes numulus (Stoll). 12. Curvipes rotundus (Kramer). 3. Curvipes alzatei Dugés. 13. Curvipes debilis n. sp. 4. Curvipes exilis n. sp. 14. Curvipes guatemalensis (Stoll). 5. Curvipes pugilis u. sp. 15. Curvipes Reighardi n. sp. 6. Curvipes turgidus n. sp. 16. Curvipes obturbans Piersig. 7. Curvipes triangularis n. sp. 17. Curvipes inconstans n. sp. 8. Curvipes constrictus n. sp. 18, Curvipes setiger n. sp. 9. Curvipes spinulosus n. sp. 19. Curvipes crassus n. sp. 10. Curvipes fuscatus (Hermann), Unfortunately the collections have none of them been made 208 ROBERT H. WOLCOTT throughout the year in any one locality, and it has been difficult in some cases to recognize the males and females of the same species, coming in certain instances from widely different localities. But every precaution has been taken to avoid the possibility of associat- ing together two sexes not belonging to one and the same species, and it is believed by the author that the characters which have caused the specimens to be associated as they are sufficient to reduce to a minimum the possibility of any confusion from the above conditions. In almost every species resemblances in form of palpus, the character of the papillae, the number and distribution of swimming-hairs, or form of the segments of the legs, and the characters of the genital area are sufficient to show the relationship between the sexes. In the descriptions of the species that follow repetition of the common characters is, so far as possible, avoided, and for brevity the following abbreviations are adopted: ep. for epimeron; seg. for segment; while the successive segments of the palpi from the base out are numbered from 1 to 5, the successive epimera, beginning with the anterior, I to IV, the successive legs by corresponding num- bers, and the segments of the legs by the numbers 1 to 6, beginning with the one nearest the base. In all cases the length of a segment is the length of a straight line joining the middle points of two lines connecting the extremities of the extensor and flexor margins. In the measurement of the palpi the length includes the claws at the tip, but in the case of the legs the length of the distal segment is exclusive of the claws or of the spine on the flexor side at the tip. Although in the collections numerous immature individuals are present, in this paper all reference to preparatory stages is avoided. II. DESCRIPTIONS OF SPECIES 1. Curvipes coronis n. sp. A species, of which the female is characterized by the possession of numerous acetabula imbedded free in the wall of the body, these forming a group approximately circular in outline, around the outer margin of which the acetabula form a distinct ring. The single female specimen of this species, collected in Saginaw Bay, Mich., is 1.254 mm. long and 1.079 mm. wide. It is broadly ovate with the anterior margin slightly produced in front of each eye THE NORTH AMERICAN SPECIES OF CURVIPES 200 and excavated between them, and the surface shows the usual fine striation. The eyes are 0.349 mm. apart, while the short, straight antenniform bristles are separated by a distance of 0.317 mm. The palpus (pl. I, fig. 2) is 0.821 mm. long, and of this the differ- ent segments make up the following percentages: 8, 27, 10, 37, 18. Seg. 1 is one-half as broad again as long ; seg. 2 somewhat less broad than long; seg. 3 much broader than long; seg. 4 rather slender, tapering, and slightly dilated in the middle; while seg. 5 tapers de- cidedly from the base to the tip. On the flexor side of the fourth segment are two papillae, one of which is very long, the other half its length, set obliquely and bearing short hairs. The same seg- ment has a short curved claw at the tip of the flexor margin. The tip of seg. 5 bears three moderately short curved claws. The spines on the palpus are more numerous than in any other species exam- ined. Seg. 1 has the usual small spine at the tip of the extensor margin ; but on seg. 2 there are, instead of the usual three on the inner surface, three in a row near the extensor margin, one a little out of line near the distal end, and one at the tip of the extensor margin, while on the outer surface, instead of two, there are two close together near the extensor margin, one near the middle of this surface, and two at the distal end, side by side. Seg. 3 has the usual number, while seg. 4 has numerous short hairs borne on small papillae. The epimera (pl. XXIX, fig. 1) are widely separated across the median line, II and III being nearer together but still with a con- siderable interval between. The legs are proportionately long, the first exceeding the body length by nearly one-fourth. The individual segments are, in order of length, 5, 4, 6, 3, 2, I, except that in the first leg 6 is longer than 4, while in the last leg 4 is longer than either 5 or 6. No particu- larly noteworthy details of structure are present. Segs. 4 and 5 of leg I possess two and five long hairs respectively, and the corresponding segments on leg II bear six and eight hairs, while on leg III are eight and eleven long hairs, and on leg IV seven and twelve swimming-hairs on the same two segments. The genital area (pl. X XIX, fig. 1) is bounded in front by a long, transversely placed chitinous plate similar to that in C. turgidus. The median area is broadly elliptical in form and the acetabula are distributed over two lateral areas nearly circular in outline, their 210 ROBERT H. WOLCOTT posterior margin extending behind the posterior margin of the median area by a considerable distance and leaving between the two a considerable space. Within these lateral areas are 23 and 28 acetabula respectively, imbedded in the wall of the body. They are, for the larger part, arranged in a row about the outer margin, while a few are scattered over the center of the area. Measurements of the single female specimen give the following figures: BOY, (eso icite nce wig eye late bis Sie eeu teny tle seine Shack atone RUAAN aa ee 1.254 mm Wipe Do hus oisshcharetxiclicgatB alo dale wie Hiern Bre AS Rak) ce ainhe ale DOPED OA Re 1.541 mm eee UM Se LL Lisi t cic cite imclale joahatales de Gite oie Mate aleve eye ae ee 1.766 mm. Wee UT ae so iatacese es clclnie exe's px alee go'v levabiel shai quagelos sie erone ele Ln er 1.805 mm. DSW ris cla aretesaisheidtie ss cinien he Clana Ae devoreres sethiie biel nest Soy oe ee ee 1.997 mm. WATS! ole ve aie niece bela. Seok Saute cess eelae kon cian eee 0.821 mm. Type retained in the author’s collection. The single female from which this species is described was col- lected in Saginaw Bay, Lake Michigan, August, 1895, by J. B. Shearer, of Bay City, Mich., and was apparently of a red color when living. The name is bestowed in allusion to the curved row of acetabula about the group of such structures, which are imbedded free in the body wall. From all the described species of which the latter state- ment is true, the present specimen seems to be quite different. 2. Curvipes exilis n. sp. A species characterized especially by the excessive length of the palpi, which are also very slender proportionally, and the fourth segment of which is remarkably contracted at the base. It is of medium size, the body elliptical in form, five-sixths as wide as long and approximately the same in height, the highest point being in front of the middle of the body. Measurements of several speci- mens from Lake St. Clair preserved in picro-sulphuric and corrosive sublimate solutions give a maximum length for the male of 0.968 mm., a minimum of 0.825 mm., and an average of 0.87 mm., while the single female measured possessed a length of 0.92 mm. The body is marked by proportionately coarse, irregular lines on its surface. The eyes are situated a little back from the margin, the distance between them being equal to somewhat less than one- third the length of the body. They are black in color. THE NORTH AMERICAN SPECIES OF CURVIPES 2iI The maxillary shield is broadest anteriorly, with rather straight sides which converge posteriorly, and a rounded posterior margin, while the ancoral process is rather short and broad. The mandibles, which are almost miniatures of those of the next species, are long with uneven outline, the dorsal margin convex distally and concave proximally, forming together with the posterior margin a relatively short and blunt process; claw long, slender, rather straight, more sharply curved at the tip, where it is finely ser- rate, and much exceeding the dorsal margin. The serrations are directly backward and limited to the terminal portion of the claw. The length is 0.312 mm. in the case of a specimen about 0.825 mm. in length. The palpus (pl. XXIX, fig. 4), which is alike in both sexes, is very long, nearly equaling the length of the body, and relatively slender, though thicker than the first pair of legs. The measurements of the successive segments give the following percentages of total length: 7, 28, 13, 34, and 18. Seg. 2 has a straight flexor margin and an extensor very slightly convex, its greatest thickness being only equal to about four-sevenths of its length; seg. 3 is barely longer than thick; while seg. 4 is nearly five times as long as thick at the base. The latter segment is much contracted close to the base, where it is the narrowest, while it is widest about the middle. Seg. 5 tapers very rapidly from the base to a distance equal to about one-fourth of the total length, after which the diameter diminishes very gradually to a tip which is rather squarish and bears in addi- tion to three small claws at the end one small one just behind them on the extensor margin. The papillae on seg. 4 are oblique to one another, the distance between their bases measured along the margin of the palpus being somewhat less than one-third the distance from the base to the first and a little over two-thirds the distance from the more distal of the two to the base of the spur at the tip of the segment. The outer of these two papillae is the longer and is equal to one-third the width of the segment at that point. The spur at the tip of this segment is prominent and shelf-like, containing a short, straight claw. The epimera cover a large part of the ventral surface and are separated from one another by a considerable interval in the case of the female, but are approximate in the case of the male. The posterior process on ep. 4 is short and blunt. 212 ROBERT H. WOLCOTT The legs are long, even the first exceeding by about one-third the length of the body; II and III are approximately equal and con- siderably longer than I; while IV is again much longer than either of the two preceding. They are abundantly supplied with spines, and seg. 5, which is first dilated and then contracted at the outer end, is furnished with a conspicuous double row of stout spines on the flexor surface. The distal end of this segment exhibits a curious modification, the anterior side being produced and forming several teeth. There are in the case of the male long, slender hairs on segs. 4 and 5 of leg II, seven on the former and five on the latter seg- ment; leg III also possesses, on each of segs. 4 and 5, about five or six swimming-hairs; while on seg. IV 4 are three and on seg. IV 5 six. In the female, leg I possesses six long hairs on the posterior side of seg. 5; leg II, six and eight on segs. 4 and 5 respectively ; leg III, seven or eight and nine on the same segments; and leg IV, two on seg. 3, five swimming-hairs on seg. 4, and seven on seg. 5. The distal segment of leg III in the male (pl. X XIX, fig. 7) is long, moderately slender, slightly curved, and longer along the flexor than along the extensor margin; its claw is short and weakly curved. On seg. IV 4 (pl. XXIX, fig. 3) there are, proximad of the excavation, two rows of three stout spines each and two spines be- tween these rows; distad of this excavation are four in an oblique line leading to the swimming-hairs. The genital area in the male (pl. X XIX, fig. 8) shows a fusion of the chitinous plates with the fourth epimeron. The genital cleft is short and surrounded by a seminal receptacle in the form of a depres- sion which is elliptical in form and with an even rim. In the median line directly behind this opening there are no acetabula, but, begin- ning with a row of the acetabula at either side at this point, there may be counted a total of 45 to 50 on each side. The chitinous plates on which these rest are long, tongue-like, and, owing to the form of the posterior margin of the fourth epimeron, show an ex- cavation of the anterior margin, while the outer and posterior mar- gins are evenly rounded. In the case of the female (pl. X XIX, fig. 5) the much longer gen- ital cleft is surrounded by an elongated, elliptical area flanked on either side by chitinous plates which are in contact with it along the posterior half of either lateral margin, but which do not meet caudad of it. The tongue-like chitinous lateral plate extends to a point THE NORTH AMERICAN SPECIES OF CURVIPES 213 even with the posterior spine on the fourth epimeron. In the male the chitinous ring surrounding the opening is connected with the genital area; while in the female it lies some distance behind this and free in the wall of the body. Measurements of two specimens give the following figures: MALE FEMALE BES CMMASM ES SMEG sah) aifeVy Shania; Sich cya aps eb opeyerar etetaver Shenae ala APA ote e 0.825 mm. 0.920 mm. SSM eyo fah a a/5.5 ba oySie’'S ats} 5\ 5) ainin’ o,aiotacthars a) & Siniioe teh diatal aerate 1.056 mm. 1.205 mm. AE ee em Leateyts cya rereore s ccaso seialai steveveleyavaiclarstehe wise aad et che: evel aerate 1.104mm. 1.310 mm. Re area etc nig Mee alere cers seis aha eve/eia olehein aie elo reratelels 1.066mm. 1.339 mm. SSO yerciavcte ialels 2eohiale oe ae ae eR ee Sates Brae Soe ante 1.248 mm. 1.546 mm. Micaela rn sie 3 aretat erst eiater actaraline Mot eioonicroee Odi Dims eee as eneecl EZEREES EIS Wavy ctastai cat ss) a) scaraist ah wie terota: Sav ere velave heen uiTaks Wavedele oraters 0.696 mm. 0.854 mm. The color of this species, as briefly noted in field notes taken at Lake St. Clair in the summer of 1893, was “transparent with a brownish tinge; with brownish yellow patches and Y-shaped mark of white dotted with carmine ; brownish red eyes; and legs that were blue with the terminal joints brownish yellow.” The types are retained in the collection of the author; co-types have been deposited in the collection of the Zoological Laboratory, the University of Nebraska, and in the United States National Museum. Specimens of this species were taken at Lake St. Clair in the summer of 1893, sixteen males and four females altogether. One male taken at Wray, Col., November 16, 1901, is also put here, being apparently identical. The specific name is bestowed in allusion to the slenderness ot the palpi, which are also longer than in any described form, and which serve to distinguish this species from all others hitherto known. The form is in length of palpi and in other details similar to the next species but is readily distinguishable. 3. Curvipes pugilis n. sp. A large species similar to C. evrilis in palpal characters, but char- acterized by the curvature of the distal segments of the legs. It has an elliptical body, uniformly arched, highest a little behind the middle with a prominence in front of each eye and a slight excava- tion between them. The breadth is about equal to four-fifths the length and the height equal to nearly nine-tenths of the same. Meas- urement of several males and females from Lake St. Clair, preserved 14 214 ROBERT H. WOLCOTT in corrosive sublimate and picro-sulphuric, give a length, for the males, varying from 1.25 mm. to 1.5 mm., the average being 1.42 mm. ; for the females varying from 1.5 to 1.7 mm., the average being about 1.58mm. The body is marked by very fine wavy lines over the surface. The eyes are relatively small and moderately distant from the anterior margin, while the distance between them is propor- tionately rather great. The antenniform bristles are extremely small and inconspicuous. The maxillary shield and the ancoral process, owing to the straightness of the sides of the former and the breadth of the latter at the base, form together a roughly triangular mass, truncate at the tip. The examination of the mandibles of several males shows them to be similar to that of the preceding species, though the claw is longer, larger, and more strongly hooked. The serrations on the claw are small, gradually disappearing toward the base. In a specimen the length of which was about 1.3 mm. the mandible was found to be 0.405 mm. long. The palpi (pl. XXIX, fig. 9) are long, being, in a male, six-sev- enths the body length. The general form is slender, though they are thicker than leg I. Seg. I is proportionately short; seg. 2 rather long and with a maximum thickness equal to about six-tenths the length; seg. 3 thicker than long; while seg. 4 exhibits a maximum thickness of only one-quarter its length. Seg. 5 is long propor- tionately and tapers gradually from the base to the tip, where it is somewhat less than half the thickness of the base. The papillae and spine on seg. 4 are similar in position and form to those of C. exilis, but are all noticeably smaller ; while there are also several small hairs in addition on the outer surface; seg. 5 has three small, curved claws at the tip, and an additional rudimentary one on the flexor margin behind the tip. The epimera cover a considerable portion of the ventral surface, are in the female separated by a moderate interval, and in the male are close together, I and II crowding the maxillary shield. The posterior projection of ep. IV is rather long but bluntly rounded. The legs are long, even the first being considerably longer than the body, while the fourth is more than half as long again. The length of the individual segments is, in each leg, in order, beginning with the longest, 5, 4, 6, 3, 2, 1. Seg. IV 6 is, however, proportion- ately lengthened in the female; III 6 is shortened in the male; and THE NORTH AMERICAN SPECIES OF CURVIPES 215 IV 4 also shortened in the latter sex, being shorter than IV 6. In thickness the segments gradually decrease from the base out, except that in the first three pairs of legs seg. 5 is much thickened, ex- ceeding in thickness in I and II all segments but the basal, and is closely beset with coarse spines on the outer half of the flexor mar- gin. The distal segment of the first three legs is peculiar in being strongly curved (pl. XXIX, fig. 11), this curvature being greatest in the case of the first pair of legs. The distal end of seg. 5 is also peculiar in the possession of a terminal expansion on the anterior side similar to that seen in C. exilis, extending, in the case of legs I and II, considerably beyond the base of seg. 6; this is less prom- inent on leg III and not developed on leg IV. The claws are small and possess two sharp, curved points and a narrow basal expansion. In the case of the male, seg. III 6 (pl. XXIX, fig. 10) is somewhat curved, slightly thickened at the outer end, and with a very weak claw with strongly curved tips. Seg. IV 4 shows, on the anterior side, a row of seven spines along the flexor margin and about five, small but of varying length, along the extensor margin; while on the posterior surface there are, proximad of the excavation, about twelve or thirteen spines increasing in size distally; in the excava- tion, three short, blunt, slightly curved spines toward the extensor margin and one large one on the flexor margin; and distad of it three stout spines in a row, and in line with these, at the tip, seven swimming-hairs. Aside from these the number of hairs is as fol- lows: On II 4 and II 5, six and eight respectively ; on III 3, two at the tip; on III 4, nine; on III 5, ten; and on IV 5, ten swimming- hairs. In the female there are seven long slender hairs on the pos- terior side of I 4, seven on I 5, seven on II 4, ten on II 5, two on III 3 at the tip, ten on III 4, twelve on III 5, two on IV 3, and ten each on IV 4 and IV 5, those on IV being swimming-hairs. The genital area of this species exhibits a length from the ex- treme anterior end to a line tangent to the posterior margin of the lateral chitinous plates of one-half that of the extreme breadth. The genital cleft is short, and the two lateral chitinous plates are separated by a considerable interval behind it in the case of the female, while in the case of the male (pl. XXIX, fig. 12) the two plates approach each other closer and the chitinous anal ring is fused with them. Each lateral plate is tongue-like with the anterior margin excavated, and the outer and posterior more or less 216 ROBERT H. WOLCOTT evenly rounded. Each possesses from 60 to 70 acetabula, of which two are noticeably larger than the rest and are situated in the mid- dle of the plate. In the male the plates are fused with the inner end of the last epimera. MEASUREMENTS MALE FEMALE BOthy.vis ui Saehanie eee meth isvatticr trek wees iw wie miele eas ebee eles 1.270mm. 1.555 mm. DST este ocsversie eisicte ecane onsets leaceote etal weeaedrace Rese aren 1.656mm. 1.776 mm. Led) I eat RA SSUES Eee SSI SHSM a SCS An HbaisioG so 1.781mm. 1.896 mm. TOE. ys sete teanvs toaster isis wee etc aatas eae ial So iets oh Sante Ie 1.723 mm. 1.982 mm. PST Me ee ele ences aE octet cia cia ieke fete aieiere CIE 1.934 mm. 2.198 mm. PALS hs Swilguse sins set Swe Sate ase pals wae es aw hee Rte 1.066mm. 1.094 mm. MANGE, sie o's facts ied vialcit ic, ose bibs orealinnc oem etomere 0.381 mame Ny cine omisiel a einielaia painiai= n!ainiwialn'sl=inin/aie 0.216 mm. Extreme breadth of genital area (approximately)................. 0.550 mm. The type is retained in the author’s collection. The single female specimen of this species was taken at High Island Harbor, northern Lake Michigan, August 18, 1894. C. medius resembles C. rotundus and C. disparilis Koenike, but differs from either in the number of acetabula and in minor struc- tural details. The name refers to its position, in reference to details of struc- ture, between C. rotundus and C. disparilis. g. Curvipes rotundus (Kramer) _ Nesaea rotundus Kramer, 79 ; 12, pl. I, fig. 6. Curvipes rotundus Piersig, 97; 118; pl. IX, fig. 19. Piona rotunda Piersig, 1901; 259. The body of this species is broadly elliptical, evenly rounded anteriorly and posteriorly, and very high, its dorso-ventral diam- eter being equal to or slightly greater than the width. There is practically no constriction behind the eyes, and the body is highest just behind the middle. The surface is marked by fine wavy lines, and all chitinous structures are rather heavier than usual in this genus. The males are about 0.75 mm. long, the females about 0.9 mm. The eyes are quite close together, the distance between them equal- ing one-fourth of the length of the body, and are slightly removed from the anterior margin. The antenniform bristles are slender, sharply-pointed, straight, and rather short. The mandibles (pl. XXXII, fig. 40) are similar to those of C. Reighardi in general shape, but the dorsal margin is somewhat more markedly excavated. The claw is more strongly curved, its tip surpassing the level of the dorsal margin, while the serration on 232 ROBERT H. WOLCOTT the flexor margin is finer and occupies the outer half. The total length of the mandible in a specimen about c.g mm. long is 0.221 mm. The length of the palpus (pl. XXXII, fig. 41) is somewhat less than one-half the total length of the body. The various segments form the following percentages of this length: 6, 27, 14, 36, 17. Seg. I is, as usual, broad; seg. 2 is slightly narrower than long with a convex extensor margin and a still more convex flexor margin ; seg. 3 is wider than long; seg. 4 is rather narrow and slightly ex- panded in the middle and with the extensor margin slightly convex ; while seg. 5 is markedly contracted before the tip, where it is about one-half the diameter that it is at the base. In the male the pro- portionate length of the segments is the same, but the palpi are relatively a little heavier and the papillae on seg. 4 more prominent. The two papillae on seg. 4 are about opposite and markedly elon- gated, equaling more than half the thickness of the segment and bearing short spines. The spur at the tip of this segment is very prominent, while seg. 5 bears not only the usual three claws at the tip, but also, close behind the claws, a short stiff hair on the ex- tensor margin and another on the flexor. The epimera cover a considerable portion of the ventral surface, being separated in the female by a moderate interval, and in the male being in close apposition. The legs of this species show a considerable difference in length between the first and last, and all but the first exceed the body length. There is, in the female, on seg. I 4 one long slender hair at the tip and also one at the tip of seg. I 5; on leg II are two or three on the corresponding segments; seg. III 4 possesses four swimming-hairs, and seg. III 5, five; while the corresponding seg- ments on leg IV possess the same number. The last segment of the anterior three legs in the female are broadly club-shaped, while the distal segment of the fourth leg of the same species is relatively small and slender. The claws are large, rather delicate, and the two outer points slender and sharp. The legs of the male are pro- portionately heavier than in the female and the distal segment, espe- cially on leg I, shows a tendency to be curved ventrad. Seg. III 6 (pl. XXXII, fig. 43) is rather stout, broader at the base, slightly curved, and bears a claw which is long, straight, and sharp. The segment preceding this is considerably elongated. Seg. IV 4 shows THE NORTH AMERICAN SPECIES OF CURVIPES 233 the characteristic form and possesses proximad of the excavation seven shorter and two longer spines, in the excavation on the flexor side one long spine, and distad of it two spines on the posterior sur- face and one on the flexor margin, together with three swimming- hairs at the tip and one spine toward the extensor margin from them. Segs. III 4 and IV 5 each bears four swimming-hairs. Each half of the genital area in the female (pl. XXXII, fig. 42) is longer and more pointed than in C. Reighardi, while the lunate plate is not only heavier but broader, and the acetabula cover less of its surface. There are in this sex from twenty-two to twenty-six acetabula on each side, of which three or four are imbedded free in the body wall; there are six spines at the anterior end of the plate, several scattered along the outer margin, and four or five at the pos- terior end. In the male this genital area is similar in form with a shorter genital cleft, at the bottom of a shallow depression and sur- rounded on all sides by an even margin, while the tongue-shaped chit- inous plates contain each thirty or more acetabula. These plates are fused anteriorly in this sex with the inner end of the fourth epi- meron. The anal opening is also surrounded by a relatively heavy chitinous ring and is situated close behind the genital area. Measurements of typical specimens are as follows: MALE FEMALE EMRE PILE EMLCLE 57.52). 3 2 oie sie ss) as Saleem eels ee came tae neta mele 0.900 mm. ESM ERCP Peay ane baal sieeisicisrecceleca/a dis) olatal oustslele; « l:ofer se 0.629 mm. 0.854 mm. ge UO eerste ease iraiore! LC Oe een renee Sera ec rr ei ac pesca, ccc oy ce 0.974 mm. eM clea ain siz csatane/aio oo « susie cab. egetaa'p Goan) ot yall tek ofa eretaeel cae 0.355 mm. Types of this species are retained in the collection of the author and co-types are also deposited in the collection of the Zoological Laboratory, the University of Nebraska, and in the United States National Museum. This species, of which only females have been taken, has been col- lected at the following localities : Four specimens at Cranberry Lake, Woods Hole, Mass., at the beginning of August, 1900; one speci- men at High Island Harbor, northern Lake Michigan, August 18, 1894; three specimens irom ponds at Columbia Mo., August, Igol ; five specimens at Circle Lake, Decatur, Neb., June 7, 1899 (Charles Fordyce) ; six at Slidell, La., August 18, 1901, and October 19, 1901 (E. Foster) ; also six from a pond in Audubon Park, New Orleans, La., August 11 and October 13, 1g01 (E. Foster) ; and specimens have also been received from the Illinois State Laboratory of Nat- ural History collected near Havana, Ill. The name refers to the variability of the species in regard to the character of the genital area, hardly two specimens agreeing in this regard. It is similar in certain respects to C. rufus (Koch), C. pauciporus Thor, and C. circularis Piersig, but comparison seems to show it quite distinct. 14. Curvipes setiger n. sp. A species characterized by the form and structure of the genital area and by the very long antenniform bristles. This species is of an elongated elliptical form with a depression between the eyes and a very slight constricticn behind them. The length of the single male in the collection is 0.698 mm., its width 0.540 mm. The females vary from 1.022 mm. to 1.095 mm. in length, the average being 1.063 mm., while the average width is about three-fourths of the length. The surface is finely striate. The antenniform bristles are extremely long, being in the single male 79 » in length and in one female measured 103 » long. They 244 ROBERT H. WOLCOTT are slender, sharply pointed, and slightly curved upward. The dis- tance between them is about five-sixths of that between the eyes. These latter are large and only a trifle over one-third as far apart . as the body is wide. The palpi are rather short, being noticeably less than half as long as the body. The percentage of length of the segments is as follows: 10, 28.5, 14, 33.5, 14. In the male (pl. XXXIII, fig. 54) the flexor margin of seg. 2 is nearly straight and the extensor con- vex, while seg. 4 is considerably dilated. The papillae on seg. 4 in the male are six in number, the two nearest the base being the longer, the shortest pair situated farthest distad. Each of the six bears a hair which in the case of the longer papillae is inserted, not at the tip, but below and behind it. The fifth segment is curved ventrad and is squarely truncate at the tip, where it bears three claws. The palpus of the female (pl. XX XIII, fig. 52) is similar to that of the male except that seg. 4 tapers nearly uniformly from the base to the tip and the papillae are very small and two in number. The hairs they bear are situated in a position similar to that of those of the male. The two anterior pairs of epimera in the male are wide apart, the other two pairs in close apposition, the length of the inner mar- gin of the two latter combined being 0.174 mm. In the female the two posterior pair of epimera are separated by a considerable space. The legs are of medium length in the male, the first being approxi- mately the length of the body, the others exceeding it, while in the female both I and II are shorter than the length of the body, III is equal to it, and IV but slightly exceeds it. The different segments are in length, in the case of the female and in order beginning with the longest, 6, 5, 4, 3, 2, I, except in the fourth leg where both 5 and 4 exceed 6. In the male the order is the same except that in leg III segs. 5 and 4 both exceed 6. In the female there are on segs. I 4 and I 5 two very fine hairs; on segs. II 4 and II 5, four or five and eight respectively ; on seg. III 3 three long hairs, and on III 4 and III 5, seven or eight and ten swimming-hairs; while on seg. IV 4 and IV 5 are six and nine swimming-hairs respectively. In the case of the male practically the same number of long hairs are found on the first two legs, but only one or two on seg. III 3, and no swim- ming-hairs on segs. III 4 and III 5, while on segs. IV 4 and IV 5 are three and nine swimming-hairs respectively. Seg. III 6 of the THE NORTH AMERICAN SPECIES OF CURVIPES 245 male (pl. XX XIII, fig. 53) is of about uniform thickness through- out, is curved ventrad, is squarely truncate, and has a small claw on the extensor side. Seg. IV 4 (pl. XXXIII, fig. 55) possesses numerous spines proximad of the excavation, one on the extensor and three toward the flexor side within it, and a row of four coarse spines distad of it, together with the three swimming-hairs. The tip of the segment is strongly produced. The genital area of the male (pl. XXXIII, fig. 51) presents a seminal receptacle broadly elliptical in form with the longest diam- eter transverse. On either side this is flanked by a plate which is nearly straight along the anterior margin, somewhat convex along the posterior margin, of nearly uniform width, and squarely trun- cate at the tip. It bears about eleven acetabula. The genital area of the female (pl. XXXIII, fig. 56) exhibits an elliptical median area flanked by two or three chitinous plates arranged in an irregu- lar manner, but occupying such a relationship to one another as would be a result of the breaking apart of a lunate plate of a form similar to that characteristic of the group to which C. rotundus be- longs. The anterior of these plates bears one or two acetabula and about eight hairs in a row around the anterior margin. Behind this is in some specimens a plate bearing one or two acetabula, while still further posteriad is a plate transversely placed bearing about eight or ten acetabula. There is also, as a rule, a large acetabulum situated within an excavation at the posterior end of the last plate, making a total altogether of from nine to twelve acetabula. Measurement of specimens gives the following figures: MALE FEMALE Bo Ocdsyaete ects 2, cfs) ny sias2)s)esier <1eieiove stateiayee seh sratyacches aero 0.698 mm. 1.066 mm. ILS? Lt 2 GSAS OR OOS CE CREE EIGER Cea eres Ie 0.696 mm. 0.921 mm. Hea OTe tert Ray otc) ota fat Flas (ale) s/s ce: shells, sis Sis iosela\) arto s lossless 0.787 mm. 1.C08 mm. ESM Merry ces raiere seit eialetersicie.e ciiste velsistel revs aide eres svalase esp 0.734 mm. 1.070 mm. Tarr) JIN Ghee Gs pad eek ae on 0.787 mm. 1.133 mm. al TASH sarcterev ei creche sieiists eve eleva veep whet She a aos eels ak taller Sev euerers 0.336 mm. 0.408 mm. The types of this species are retained by the author and co-types of female specimens deposited in the collection of the Zoological Lab- oratory, University of Nebraska, and in the United States National Museum. C. setiger has been collected in only one locality, a spring-fed pool in Monroe Canyon, Sioux County, Neb., May 28, 1899, where one male and ten females were taken. 16 246 ROBERT H. WOLCOTT The name is given in allusion to the length of the antenniform bristles, which exceed by considerable not only that of any other of our American species but that of any other species hitherto dis- covered. It closely resembles at first C. inconstans but differs in the follow- ing particulars: The antenniform bristles, as just stated, are very much longer; the proportionate lengths of the segments of the palpi and the character of the fourth segment are both different ; the num- ber of swimming-hairs on the legs is much greater; the relative lengths of the segments of the legs is also much different; while, in the case of the genital area, although the number of acetabula is not diagnostic, the character of the chitinous plate that bears them is different. The species resembles the same three European species referred to in connection with C. inconstans, but seems entirely dis- tinct, on comparison of details of structure. 15. Curvipes crassus n. sp. A heavily built species with very marked structural features. From the examination of four males and a single female of this species the form appears to be elliptical, with a moderate flattening between the eyes. The cuticula is very thick and marked by numer- ous close raised ridges which are of uneven height, so that in oblique view the surface looks almost as if covered with evenly distributed, low papillae. Of the four males the longest is 0.635 mm. in length and 0.508 mm. in width, the average being 0.605 mm. and 0.518 mm. respectively. The female is 0.793 mm. long and 0.659 mm. wide. The body is moderately high and quite evenly convex dorsally. There is below and slightly within each eye a flattened, blade-like antenniform bristle of considerable length arising from a low pa- pilla ; in the largest male referred to above it is 30 « long. The eyes are large, near to the margin, and separated by a distance equal to a little less than one-third the average breadth of the body. The maxillary shield is unusually narrow and the ancoral process relatively short and strongly hooked. The mandibles are rather broad, expanded posteriorly, and with a rounded angle between the dorsal and posterior margins, while the claw is relatively small and angulated, tapers to a slender but blunt point, and shows no serra- fion. The palpi (pl. XX XIII, figs. 59 and 60) are stout, heavily chit- THE NORTH AMERICAN SPECIES OF CURVIPES 247 inised, and very characteristic in form, with a total length a little less than half that of the body. The following numbers represent the percentage lengths of the segments: I0, 31, 16, 27, 16. Seg. I is not quite twice as broad as long; seg. 2 is about the same in thick- ness as in length; seg. 3 is somewhat thicker than long; and seg. 5 is rather narrow at the base, where it is less than half as wide as long, and tapers to a tip equal in width to about one-fourth the length, and which bears three small distal claws. Seg. 4 is strik- ingly modified, a very prominent projection of its ventral margin bringing the distal portion of this margin almost in line with the distal end of the segment and at a right angle with the proximal portion. The diameter of the segment at the distal end becomes thus nearly twice that at its base and equal to three-fourths its total length. On this distal portion of the flexor margin are placed three pairs of blunt conical papillae, and outside the line of these, between the first two proximal pairs, is another papilla on either side, mak- ing eight altogether. All of these papillae bear hairs, which are on the two proximal ones moderately long, on the rest short and slender. The palpus of the female is similar to that of the male in size and proportions of segments, but the distal portion of the flexor margin of 4 bears only two pairs of papillae, each one with a short hair. The claw at the distal end of this margin is in both sexes represented simply by a blunt projection. The epimera in the male are all in contact except that between the inner ends of the first two pairs is a small area not covered by them. Over the epimera are seen coarse parallel striae. The process on the posterior margin of ep. 4 is very long and sharply pointed. In the female the epimera are similar in form but the four groups are sep- arated by a narrow interval. The legs are rather short, the first pair in the male being only slightly shorter than the body and the others all exceeding it, while in the female the last two exceed the length of the body; they are rather stout, the chitinous covering being here, as elsewhere, much thickened. In leg IV the four basal segments are much stouter pro- portionately than the others. The segments increase in length also from the base outward, though in the male 6 is shortened so that it is shorter than 4 and in leg IV it is still exceeded by 5, while in the female the same is true in legs III and IV. In legs I and II seg. 5 has a convex flexor margin and seg. 6 is very markedly expanded at 248 ROBERT H. WOLCOTT the tip (pl. XX XIII, fig. 57), while the spines are few. Segs. 4 and 5 also show a distal expansion on the anterior side. In leg III of the male (pl. XX XIII, fig. 58), 5 is narrow at the base and dilated at the distal end, while 6 is very short, squarely truncate at the end, its flexor margin straight and its extensor margin strongly convex at the base, beyond which it is parallel to the flexor. The claw is small and straight. In leg IV of the male segs. 5 and 6 are noticeably slender in contrast to the marked dilatation of the basal four segments, already referred to, and seg. 6 is slightly dilated to- ward the tip. The same dilatation of the distal segment characterizes the legs of the female, though it is in much less marked degree. Seg. 4 of this leg is broadly expanded and with a deep excavation on the posterior surface, proximad of which are many stout spines, and distad of which is a row of three short, stout spines, two swimming-hairs, and a very heavy spine at the tip on the flexor side. In the male are three long hairs in a row at the tip of seg. II 5, one at the end of seg. III 4, the two swimming-hairs on IV 4, and six on IV 5; while in the female seg. I 5 bears one slender hair, seg. II 5 four swimming-hairs, segs. III 4 and III 5 two and five swim- ming-hairs, and segs. [V 4 and IV 5 three and five respectively. The genital area is in close contact with the posterior epimeron in the male and extends around on the outer portion of the angle on the posterior surface to beyond the outer end of the epimeron. There is no seminal receptacle and the genital cleft is very short, being 41 » long. In the female the two genital flaps form a broadly elliptical area, from the posterior half of either lateral margin of which project two long tongue-like chitinous plates which are con- cave on the anterior and convex on the posterior margin and which extend to beyond the line of the angle on the fourth epimeron, while there are also one or two acetabula at the margin of the median area and midway between these plates and the anterior end of the area. Each plate flanking the genital area of the male bears 50 to 60 acetabula; in the case of the female over 70. Measurements of specimens: MALE FEMALE BOG shee ajatie) fave sousteyalsdasaep one chotebele aie lavateucate iets ede uetale (els 0.659 mm. 0.793 mm. De S Yoryesd ack oieteue era ee ein Seton re laiceie eden rater ateietebe foretole 0.619 mm. 0.682 mm. De eee aeatete tere tom Rial celalels Wren ntene eyenene tee camete ors 0.682 mm. 0.754 mm. MgO TDD Poe eR hs anevada baste terete by ats are Ate arate me Palette 0.749 mm. 0.830 mm. THOS TI ch ayesha AR aC Ra EH IU Sa 0.840 mm. 1.042 mm. Pal puss isiecs shove ete eis coe NSS Cita ON GIRARD rete 0.3860 mm. 0.365 mm. THE NORTH AMERICAN SPECIES OF CURVIPES 249 The color of specimens collected in Susan Lake, near Charle- voix, Mich., was noted in field notes as follows: “Body tinged pos- teriorly and around margin with bluish green, most pronounced at the posterior end. Anterior median portion strongly tinged with dull reddish. Eyes black. Legs a bright bluish green with terminal joints brownish.” Types retained in the author’s collection. This very characteristic species has been collected in the follow- ing localities in Michigan: Near Grand Rapids, in the summer of 1885, two males; at Lamberton Lake, near Grand Rapids, July 4, 1900, one female; at Susan Lake, near Charlevoix, August 21, 1894, three males. This species is similar to no other species of Curvipes except C. thoracifer Piersig, from which it differs, however, in all details of structure, including the characters of the palpi, the space between the epimera, the number of acetabula, the form of the genital plates in the female, etc. The name has reference to the strength of the chitinous covering and the stoutness of the appendages. III. TABLE FOR THE DETERMINATION OF THE ABOVE SPECIES FEMALES 1. Acetabula imbedded free in the wall of the body............. 1. C. coronis Acetabula in part or all on chitinous plates.......... 0.66. -eee cece ee eeee 2 2. Two genital plates, one on either side ..........- eee e eee eee e eee eee 3 More than! oftelonjeaGh, SIDE... «aye c1oyajaheasieials cp aiotcloleiei=ts) ©) vellsielatep nia etastarsplelels 12 3. The genital plate solid, its whole inner margin in contact with the me- bastellipiteal | ATCA... ssc... «ais «) 2 o:eie) a alalabeere ainie' ates ioies/=liin'ni Aejofe sh opekeia fei aha 4 The genital plate lunate, enclosing an area in which a greater or less number of acetabula are imbedded free in the body wail........... 9 4, The distal segment of the legs strongly curved .............. 3. C. pugilis The distal segment of the legs not curved... 2... 20-2 csseccecseccescences 5 Ha AcetaA pila Mum erOus,, 4) OF OW sa\2 siesslsiak cisietedefaios)cleie ol els cisiaiels!all mis/elaelelota ove 6 Aicetabulay fewer not. OVer oo.) cls oeiie (state oaryojslans ara, sdoleiolevele eicloiaveresaweleialskate 7 6. Palpus very long, nearly equaling the body in length and the fourth segment markedly contracted at the base................. 2 Cleats Palpus not over half the length of the body, fourth segment tapering, 4. C. turgidus 7. Acetabula about ten, small, on a very short plate......... 6. C. constrictus Acetabula from twenty to thirty, large, on a plate longer than broad.... 8 8. The epimera confined to the anterior half of the ventral surface, body larger, and swimming-hairs on segs. IV 4 and IV 5, five and seven MEIC ELV EBS ania a claleroie oiciala wfatero a wietntal ers for sia yaiel a ale alan 5. C. triangularis 250 ROBERT H. WOLCOTT 10. dele 13. bo The epimera covering most of the ventral surface, body very small, and swimming-hairs on segs. IV 4 and IV 5, four each ....7. C. spinulosus . Papillae on seg. 4 of the palpus long, equaling in length half the thick- ness of the segment or more .....0 0.5 60 6. Slee ones oe ons aisle eee 10 Papillae on.seg. 4 of ‘the palpus.Short......05.).0s265 css: ee cem eee eee 11 Acetabula numerous, 35 to 40 on each plate..................8. C. medius Acetabula fewer, 22 to 26 on each plate.................0.- 9. C. rolundus Swimming hairs fewer, on segs. III 4 and III 5 being three or four and five or six; and on IV 4and IV 5, three and four; two papillae on fourth.'seginent of palpus. <> .'....\..4..0..0sese. 08 ll. C. Reighardi Swimming-hairs more numerous, being on the four segments named, nine, nine, seven, and five; two small papillae on fourth segment of palpus in addition to two above ...............68. 12. C. obturbans . Distal end of the fourth palpal segment much dilated, as also distal segments of legs; chitinous parts all coarse and heavy.. 15. C. crassus Fourth palpal segment tapering, and all chitinous parts slighter........ 13 Antenniform bristles of usual length; acetabula from eleven to eighteen; swimming-hairs on segs. IV 4 and IV 5, three and four respec- EUVE LV ce tacit ts asiotecetmieists siviehs music le slceun Sei eiehe cere 13. C. inconstans Antenniform bristles very long; acetabula from nine to twelve; swimming- hairs on segs. IV 4 and IV 5, six and nine respectively...14. C. setiger MALES . A seminal pouch or seminal receptacle present...........2eecccececceess 2 Simply a depression at the bottom of which is the genital opening....... 4 . Acetabula numerous, 40 to 50 in number.............-2000- 4. C. turgidus Acetabula few, from nine to elevetl. ..... 5... 3.2 200+ +s ons nat eemeieeianen 3 Several long, slender hairs on seg. III 5 and ‘fifteen swimming-hairs on IV 5; two papillae on the fourth segment of the palpus; seg. III 6 Commucdpia-Sla Ded, nce nmieyaieiaw vise © eee pena einer 6. C. constrictus No such hairs on seg. III5 and only nine swimming-hairs on IV 5; six papillae on the fourth segment of the palpus; seg. III 6 slightly bent, not broader at the tip’ 0.12.2 oiien i series oe == 14. C. setiger . One claw at least on seg. III 6 long and straight............. sees eeee cues 5 Neither long and straight, but short and curved .......... sees sees eeeees 6 . Size small; papillae on the fourth segment of the palpus of medium length; swimming-hairs on IV 4 and IV 5, three and four respec- tively; ‘acetabula 25 to'S0?: Jo Scie oe cee wwiniem = mira 7. C. spinulosus Size medium; papillae on the fourth palpal segment long; swimming- hairs on as in the preceding; acetabula about 30....... 9. C. rotundus Size rather large; legs relatively weak; papillae on fourth palpal seg- ment short; swimming-hairs on segs. IV 4 and IV 5 three and nine respectively; acetabula fifteen to eighteen.......... 10. C. debilis . Fourth palpal segment not greatly dilated at the distal end and with two papillae. oo... och ela cele wens ones once mone iusiulsin selena Fourth palpal segment greatly dilated at the tip and with eight pa- DILAE |i oa ioe s die'e ars Sind wise eia's lero Siete e nie’ s lems aikinieletale a alana 15. C. crassus THE NORTH AMERICAN SPECIES OF CURVIPES 251 7. Distal segment of legs not strongly curved; three swimming-hairs on Siete I\W/ee Apnea b On CUI BOE IOCamWGUadn GOC dd po bouA OLE DC. Gxtilis Distal segm:nt of legs strongly curved; seven swimming-hairs on seg. TV Ae oasis, sia cidisin ela srsnain we.aie aleacs ala wax dleaeahola oiahalet aes ee amine eyes oe 3. C. pugilis In fitting the species described above into Piersig’s table (1901: 244), the following changes become necessary: FEMALES 4, Maxillary palpus much stronger than the basal segment of leg I; etc., 2. C. aduncopalpis Maxillary palpus little stronger than the basal segment of leg I; etc....4a 4a. Acetabula scattered irregularly over a considerable area, sixteen to GEwerhy im timbers. .\:.cis «\s ceness serie eevee enacts Me roe 3. C. conglobatus Acetabula more closely collected into a group with circular outline, twenty-three to twenty-eight in number ................... C. coronis BePMereiMAry Palpiis Smal CLE. e's 0.5 oiciciatn asia osetia’ esse aicielera, sat Natalee Melnlareta ees 8a Niartliiy palpus lanmes CLGs operas oyetateiciaieler ocnsn.4 we ayaiinsa are ere itehoncieie Seana me 9 8a. Genital plate circular, bearing eighteen to twenty-four acetabula, 6. C. carneus Genital plate very short, crowded against median area, bearing about PE TACE CATA stecaayegoretet sretce sat shai cs ieee atctate anal cunt altho tats Monsees C. constrictus ew several larce papillae, sete. 22's. Sire Siaie/s evbistanistdeino-aiclalatoe 7. C. uncatus With two large papillae and two very small ones beyond them, C. turgidus PEERED WOOL! clei) tis abetajale (alan lu chalgp mage deleard oiatais*ais «ay shdpatay hore epee oe 10 taaeAtcetabtla: numerous; 45) OF SO Wena cicis oa olees clei stecd see Han Ose 13a iNcetapula fewer NOt. OVeEr- DOL 5: s/svate «leeysrs oaatavere cielaeloichiove clare tar ree eee 18¢ 132. Distal segment of legs strongly curved...............000 e000 C. pugilis Distalisegment of legs not strongly carved): 2). ..0! i 56.429 atten spain 130 136. Leg III shorter than leg II; palpus about half the body length, ll. C. longipalpis Leg III longer es leg II; palpus nearly equaling the body in length, C. extlis 18c. Size large, about 2 mm.; a group of hairs on the last epimeron, along the outer portion of the posterior margin.............. 12. C. nodatus Sizenmedaumuor small snoisuch! hairsau- sce eee cee eee 13d 13d. Size medium, about 1 mm.; epimera confined to anterior half of ven- BE ARUOSTLACE < fa ks stoitretonlaraie Sine taeeiala che etary a a eo tebe C. triangularis Size small, not over 0.6 mm.; epimera covering most of ventral sur- TLCS 0 raes ob ants Wattage crac ene cree ae Ssh yt cea ie at Ale C. spinulosus 20. Each genital plate with over 50 acetabula............... 20. C. disparilis Each genital plate with 35 to 40 acetabula.................000- C. medius Each genital plate with less than 30 acetabula ..................0.002- 21 21. Papillae on the fourth segment of the palpus long........ 21. C. rotundus Papillae on the fourth segment of the palpus short................... 21a 21a. Swimming-hairs fewer; two papillae on fourth palpal segment, C. Reighardi ROBERT H. WOLCOTT Swimming-hairs more numerous; two small papillae additional on the fourth palpal segments. Void are Polels nls to deacon seca 22. C. obturbans 24, Posterior genital) plates transverse, ete...) 001/002. Go dea eee 25 Posterior genital plates. curved aati <2 de - bes if 64 * =—y vr} Pr. ag “ . ; se; £m iv. P ale - A ) wr _¥ e a tyague ape awe en) 4 ipl ihe if ert ear od Bice - Ai . ade , a & t * 2 eo 7) PLATE XXXII R. H.W. det. Fig. Fig. Fig. Fig. Fig. Fig. Fig. Fig. Fig. Fig. Fig. Fig. 110. Fig. Fig. Fig. Fig. Fig. Fig. Fig. Fig. Fig. Fig. Fig. Fig. 36. 37. THE NORTH AMERICAN SPECIES OF CURVIPES Plate XXXI Figs. 24-29. C. spinulcsus . Epimeral field and genital area, ¢; «160. . Epimeral field and genital area, 4; 160. . Segment III 6, 4, anterior side; 439. . Inner side, right palpus, ¢; 335. . Inner side, left palpus, 2; 330. . Posterior side, segment IV 4, 4; 335. Figs. 30,31. C. medius . Genital area, 2; 160. . Inner side, right palpus, 9; 260. Figs. 32, 33. C. triangularis . Outer side, right palpus, 9; 155. . Genital area, 2; 135. Fig. 34. C. Reighardi - . Claws, tip of segment III 6, 9; 625. Plate XXXII Figs. 35-38. C. Reighardt Outer side, left palpus, 9; 260. Genital area, 2, from Lake St. Clair; 260. (cof. fig. 35.) . Inner side, left mandible, 9; 335. Fig. 39. C. obturbans . Inner side, left palpus, 9; ><260. Figs. 40-43. C. rotundus . Inner side, right mandible, 2; #35. . Outer side, right palpus, ¢; 260. 2. Genital area, 9; 335. . Segment III 6, ¢; 625. Figs. 44-46. C. debilis . Segment III 6, 4; 365. . Outer side, left palpus, 4; ><260. . Genital area, ¢; from an unmounted specimen; 110. Fig. 47. C. inconstans . Outer side, right palpus, 2; 369. 255 . Epimeral field and genital area, ¢, from High Island Harbor; 256 ROBERT H. WOLCOTT Plate XXXII Figs. 48-50. C. inconstans . Genital area, 9, from Slidell, La.; 260. . Epimeral field and genital area, 9, from New Orleans, La., «100. . Genital area, 9, from Decatur, Neb.; 260. Figs. 51-56. C. seliger . Genital area, ¢; from the unmounted specimen; 160. . Outer side, right palpus, 2; 260. . Segment III 6, ¢; 485. . Outer side, left palpus, 4; 135. . Segments IV 3 to IV 5, ¢, posterior side; 260. . Genital area, 9; 260. Figs. 57-60. C. crassus . Segments II 5 and II 6, 4, anterior side; 335. . Segments III 5 and III 6, ¢; 260. . Outer side, left palpus, ¢; 335. . Outer side, right palpus, ¢; 260. PLATE XXXIII RHW, der. A NEW HYDRA By M. J. ELROD, UNIVERSITY OF MONTANA, AND MAURICE RICKER, BuRLINGTON (Iowa) HIGH SCHOOL During the summer session of the University of Montana Bio- logical Station, we found what is believed to be a new hydra. It was taken in large numbers from Echo Lake, Flathead county, Montana. It has not been found in any of the other numerous streams or lakes in this vicinity, and so far as is known no other hydra has ever been collected in the state.* The following are some of the most noticeable characteristics: The animals are conspicuous on account of their bright coral red color and large size. In fact, one can recognize them as hydrae while standing erect on the logs. A fair sample of the larger ones measured, when feeding, 16 mm. long from the mouth to the distal end. None of the tentacles of this hydra were less than 38 mm. long, measured from the mouth to the end, and the longest was 43 mm., making a total length from tip to tip of 59 mm. When feeding, the tentacles seem capable of unusual extension until they seem a mere thread, bearing noticeably large nemato- cysts, like beads strung on a string. The color is a deep bright coral red, most intense near the proximal end and seems to be distributed in chloroplast-like gran- ules as in H. viridis. It is apparently constant and may possibly be due to symbiotic algae, although indications are to the contrary. Since the waters of Echo Lake contain large numbers of a reddish Daphnia, and, thinking the question of their effect on the color of the hydra would arise, a number of the latter were taken alive and fed for five weeks upon colorless entomostraca, from Flathead Lake, at the Station laboratory. While they did not seem to thrive, no noticeable dimming of the color bodies was observed. The hydrae were found early in July, 1901. There was little time or facilities for delicate histological work, and the lack of the literature compelled us to defer more careful examination until a more convenient time. i 1Since the above has been in type Prof. R. A. Cooley reports finding a hydra sparingly in the eastern part of the state. 258 M. J. ELROD AND MAURICE RICKER The striking color, the large size, the isolation of the animals from related forms, the apparent division of the body into a stalk and an enlarged gastric cavity of about equal length, the removal of gcnads and buds beyond this apparent division, altogether seemed to make it worthy of this preliminary note. Careful histological examination will be immediately made, and should the characters enumerated, together with others which may be revealed, prove constant and new, as it is believed they will, the name Hydra corala is proposed for the species. Echo Lake, in which the hydra was found, lies in the valley close to the foot-hills, west of the Swan Range of the Kootenai Mountains, a few miles northeast of the Biological Station. It is narrow, with a total length of twelve or fourteen miles. It may be the old bed of a river which, in earlier days, flowed through the valley until dammed by a moraine. The lake now has no surface outlet, the water probably escaping through underground channels or seepage. Notwithstanding, it contains five or six species of fish and numerous species of entomostraca. In 1894 the water in the lake suddenly rose about twelve feet above its former level, submerging portions of timbered lands about the lake borders and a meadow. The water has remained at this higher level since that time. At the upper end of the lake a rancher took up his claim when the water was at the lower stage. He built a log bridge over the little stream which flowed into the lake at this point and erected some log buildings in the meadow. The rising water floated the bridge’ and came up to the top of his door and windows. It was about this bridge, attached to boards and roots of grasses growing between the logs, that the hydrae were found. Echo Lake affords a good field for further biological study. Its waters are held in by a moraine and have no connection with other bodies of water at any season of the year, unless by unknown under- ground channels. Among other interesting collections was a Polygonum which seems to have made important adaptations to its new conditions. The potamogetons were exceedingly long, and there is no doubt other evidence of a quick response to changed environment could be easily found. The further study of this lake will be undertaken by the members of the Station staff and their students next season. MODIFICATION OF SOME STANDARD APPARATUS TO FACILITATE THE WORK OF THE HISTOLOGIC AND EMBRYOLOGIC LABORATORY By SIMON HENRY GAGE, CORNELL UNIVERSITY, ITHACA,N. Y. WITH ONE PLATE (1) AN IMPROVED SECTION RAZOR WITH STRAIGHT BACK AND EDGE It seems strange that when an instrument takes on new uses it is so slow to lose distinctive features which have no significance or are harmful in the new field. There is thus a certain similarity between a machine in its evolution and an animal or plant. It took a long time to get a section razor with a straight edge for section work, and apparently no one ever thought it necessary to eliminate the compound curves of the back. In the form devised about a year and a half ago the back and edge are both straight, and as nearly as possible parallel. This makes it possible to change the position of the razor in the holder without changing the angle of the cutting edge (pl. XXIII, figs. 1,°2, and 3). The razor here presented and advocated, then, has edge and back straight and approximately parallel. The razor blade is thick so that it will not readily spring in cutting sections, and it is slightly concaved on both sides to facilitate sharpening. The writer has never yet been able to find out why razors used in histologic work are flat on one side in so many cases. (2) A RAZOR HOLDER AND SUPPORT FOR THE MINOT RIBBON MICROTOME From the cheapness and excellence of razors for sectioning and from the ease of sharpening them, they are used almost exclusively in student work, and also in research work in many laboratories. The Minot ribbon microtome is designed for a section knife of con- siderable size, and if a razor is placed in the regular holder it can be 260 SIMON HENRY GAGE moved very little from side to side. This makes it possible to use only a small part of the cutting edge, and the whole razor must be sharpened every time the small part is dull. To avoid the difficulty, a support was devised about two years ago. This consists of a strong piece of brass which rests in the knife support of the microtome. At right angles with the base-piece, on which rests the back of the razor, is a vertical back-piece against which the side of the razor rests. This is slightly narrower than the width of the razor blade, and a notch is cut out of the middle where the sections are made. A front-piece is made like the back-piece, except that it is not fastened to the base-piece. This is put against the front side ot the razor and the clamping screws of the regular knife holder press against it (see figs. 2, 3, 4, and 5). If one employ such a support with the Minot ribbon microtome, nearly the entire length of the edge of the razor can be used. It is highly advantageous, however, to have a razor with a straight edge and back (see above and fig. 1). Not only should the back be straight, but the haft of the knife should be thin enough so that the angle of the knife will not change in moving the razor. (3) AN ADJUSTABLE CLAMP FOR THE MINOT RIBBON MICROTOME In using the Minot ribbon microtome the holders for the paraffin blocks furnished with the microtome are expensive, and only three come with each microtome. Finally, the clamp to receive these block holders has very slight adjustment, so that the holders must be very accurately fitted. At the Columbus meeting it was pointed out that in a laboratory where many students work and use the micro- tome there must be many holders for the paraffin blocks. To make this possible with a minimum expense, short stove bolts were rec- ommended. These can be used as they are, or a coin like an Amer- ican cent can be soldered to the end for a larger attaching surface. From the small adjustment in the clamp for the holder many of the stove bolts could not be used without much trouble in fitting them. To avoid this difficulty an adjustable clamp was devised which will receive bolts differing one or two millimeters in diameter. Two views are shown (figs. 6 and 7). The stem which connects the clamp with the other clamp of the microtome has a long thread and a solid piece is screwed upon it. A loose piece like the first is then slipped over the screw, and finally a thumb nut is put upon the MODIFICATION OF SOME STANDARD APPARATUS 261 end to press the loose piece against the fixed piece. Holes are bored in the clamp, half the cylinder being in each. Either of these holes serves for the paraffin block holder. With such a clamp one does not have to worry about the exact size of the stem of the paraffin holder. (4) AN IMPROVED TRAY FOR HOLDING SLIDES OR RIBBONS OF SECTIONS The tray exhibited and described at the Columbus meeting proved itself so excellent on extended use that one or two defects have been overcome. The defects were two: First, the outside frame had square corners and sharp edges. The least warping or irregularity made them lock into each other so that it was not easy to pull one out of a pile, nor was it easy to return it to its place again. To avoid this all the corners and edges have been rounded. Slight irregu- larities do not now interfere with the removal or return of a tray in the middle of a pile. The second difficulty was in getting hold of a single tray when they were in a pile. This was easily overcome by adding a small screw eye. With the improvements thus indicated one has no longer the necessity of purchasing expensive slide cabinets. These answer every purpose and are exceedingly cheap, costing only about $15 per hundred for trays which will contain fifty 3 x 1 inch trays. 17 262 SIMON HENRY GAGE EXPLANATION OF PLATE Plate XXXIV Details of razor, razor holder, and support for Minot Ribbon Microtome and of adjustable clamp for Paraffin Blocks. PLATE XXXIV 7 Gil in Gaption Figure 1] Front piece of Razor Holder For Razor LABORATORY PHOTOGRAPHIC APPARATUS By SIMON HENRY GAGE, CoRNELL UNIVERSITY, ITHACA, N. Y. WITH TWO PLATES (1) A TABLE AND SCREEN FOR USE WITH A VERTICAL PHOTO-MICRO- GRAPHIC APPARATUS Much if not a majority of photo-micrographic work is done with a vertical camera like that of Zeiss. To avoid various inconveni- ences in the use of this, a small table about 50 cm. high was con- structed upon which the microscope and camera rest. This makes it possible for the operator to stand upon the floor for adjusting the camera, and to sit on a low stool for adjusting the microscope. To avoid eye-strain from the light a zinc screen was made, with legs and heavy bases to fit over the table and between the microscope and the lamp. This screen is slightly higher than the vertical cam- era, so that when the bellows are pulled out to the extreme limit one can work without the lamp shining in the face of the observer. Opposite the lamp is a perforation in the screen. This is covered by a balanced curtain so that the light is very easily shut off at the desired moment (pl. XXIV, fig. A). (2) A SPECIAL MICROSCOPE STAND WITH THE STAGE IN PLACE OF THE TUBE, FOR PHOTOGRAPHING EMBRYOS AND OTHER SMALL SPECIMENS WITH A VERTICAL CAMERA As shown in the accompanying figure (pl. XXIV, fig. B) the small vertical photo-micrographic camera is placed upon the low table as in photo-micrography. In place of the sleeve for connecting the camera to the microscope, a photographic objective of 60 to 80 mm. focus is employed, thus making an ordinary camera of it. To support the specimen and also to serve as a focusing arrangement, a skeleton stage is attached to the arm of the microscope stand. The stage proper is absent. From this arrangement the specimen 264 SIMON HENRY GAGE may be focused with either coarse or fine adjustment. For opaque objects there is a second staging below on which may be placed any desired background. For transparent objects a large mirror is in the usual place and serves to illuminate the object. For photo- graphing embryos it is desirable to know the exact size of the pho- tograph as compared with the specimen. -To determine this quickly in each case the instrument is at first calibrated. That is, a centi- meter rule is used as object, and the various sizes desired, e. g. natural size, twice, four times, five times, etc., are cbtained by using dividers and measuring the image on the ground glass and noting the exact position of the upper and lower part of the camera. Having once determined these points, the camera may be set at the one desired in a moment. Then the focusing is done with the spe- cial microscope stand. (3) USE OF THE SPECIAL MICROSCOPE STAND WITH AN ELEVATED CAMERA TO OBTAIN PICTURES OF LARGE SECTIONS, ENLARGE PHOTO-MICROGRAPHIC NEGATIVES, OR MAKE LAN- TERN SLIDES FROM THEM In photographing very large microscopic sections of embryos or of organs it is difficult to illuminate evenly the sections. To overcome this difficulty the camera is put in reverse order on the frame for the large vertical camera, and then elevated sufficiently to ensure a sky background. This will give even illumination. The specimen is placed on the stage of the special microscope stand. The focusing is then performed in the well-known way by a cord over the fine adjustment. One can focus as accurately as with a micro- scope. Objectives as high as 35 or 42 mm. may be used; or one may use a short focus photographic objective like the micro-planars of Zeiss. PLATE XXXV 4 } B A Small table and screen for the vertical Special microscope stand with stage in photo-micrographic camera. place of the tube, so that the specimen - can be focused with the fine or coarse adjustment. = 9 bh dave a eh a , 4 f ‘ ’ i A A i 14 iy : ‘ ss 7 4 F “ cS . ‘ ' o " P . ‘ 4 "1 - ga - . ‘ ‘ \ " a ¢ = ‘ ¢ os Vt I ‘ ™ . ; . ‘ ‘ : ? ! ” _ a ‘ ' / * « ’ ‘ ( — = ’ ‘ , . . * ‘ i ia ‘ » as XXXV “ 4 E PLAT ‘UOTJLUIMINT]LE JO 9.1nN0s B Se AYS dy} Suisn ‘suorjoas a81P| AigA Surdeis0joyd 10; puvys sdooso1sur yetoads YM ede po}eAag[H *payeagya Ayj1ed yng SMELLS IY} PUL WOT}ISOd UI pasIdAdI SI BIBLES JY} ‘9}a ‘sUBSIO JO SUOTIOAS s1dossois1m adie, Surydeisojoyd 10,7 ‘smopeys proav 0} Mojaq AIqe -I9PISUOS ST PUNOISYSe dy} PUL SS¥[S YIM poi1dA0d SI 91qe} Oy, “SJoal -qO 19430 pure stieiq Surydei1s0joyd 107 versio o1ydessojoyd [eo13A @ a —=== SS MOM if — © REPORT OF THE COMMITTEE ON THE SPENCER-TOLLES FUND Your committee, which was appointed at the last meeting to con- sider the possibility of developing this fund, begs leave to report that the matter has been brought to the attention of all members of this organization and other societies having an interest in the devel- opment of microscopical science; and that by virtue of the contribu- tions which have been received, and which are enumerated in the report of the Custodian it has reached the limit set at the New York meeting of twelve hundred dollars. We are glad to report that great interest has been manifested in the fund as a memorial to men who have done so much to develop the tools of the investigator and to make modern research in so many lines fruitful of great results. To each and all those, both individuals and organizations, who have contributed so generously toward the completion of this fund, it is appropriate that this Society should here formally acknowledge its thanks at the same time that it expresses its appreciation of the responsibility which the carrying out of this charge lays upon it. It has seemed appropriate to append a complete list of contributors to the fund. Although the limit set has been reached and the amount on hand will yield an income such that at least a small sum may be devoted annually for the purposes of the fund as the Society may designate, yet it is clear that continued effort should be made and certainly will result in securing from time to time additions to the total amount. In the opinion of your committee the policy of the Society should be to build up gradually an adequate fund looking toward the future possibilities of a larger income in developing that branch of science for which this organization stands. It is accordingly recommended that a standing committee be appointed to care for the fund and to develop it in a conservative manner along the lines laid down above. Officially the fund now stands in honor of the memory of Robert B. Tolles and Charles A. Spencer. With the name of Spencer, how- ever, is associated, not only the illustrious father but also his dis- 266 REPORT ON THE SPENCER-TOLLES FUND tinguished son, the late Herbert R. Spencer, a loyal member of this organization and a successful worker for the advancement of micro- scopical science. Accordingiy your committee deems it appropriate that the fund should stand as a memorial to the genius of this trio of American scientific workers, and recommend that it be desig- nated as such officially and be known hereafter as the Spencer- Tolles Fund. The legal advice which was at the command of the committee was of the best, and it seems clear that the present investment of the fund is conservative and safe. The committee would accordingly recommend that the Custodian should be directed to keep it invested as heretofore, and apply any additional moneys which have been or shall be received to increase the principal of the fund. Regarding the use of the income it may be stated in advance that the object which the contributors have had in mind, and which mem- bers of the Society have cherished, seems to be in general the en- couragement of research, especially such as depends upon the microscope for its accomplishment or is connected with the im- provement of that instrument. The committee has carefully reviewed the practice of other organ- izations in the world having trust funds of this character, in order to ascertain how these have been applied and what results have been obtained thereby. The following possibilities have thus been pre- sented for consideration : The awarding of prizes, which is a somewhat general method of disposing of the income of such funds, does not seem to be attended by results proportional to the expenditures, and the experience of this Society in offering such prizes at previous times has led mem- bers of the committee, together with such other members as they have been able to consult, to advise unanimously against the appli- cation of this fund to such purpose. It has also been suggested that the money might be placed in charge of some other society to increase the funds of such organiza- tion. We do not feel that this would meet the ideas of the contrib- utors who have given the money in charge of this Society and who will naturally look to it for the carrying out of its trust. Further- more, it is evident that the administration of such a trust will be attended with some honor to the Society charged with its execution, and this belongs by right to the organization which has raised the — REPORT ON THE SPENCER-TOLLES FUND 267 fund; therefore we are equally unanimous in advising against such a procedure. It has recently been recommended that the money should be placed in the hands of some institution to found a scholarship for educational research bearing the names of Spencer and Tolles. But quite apart from the difficulty of deciding which institution should be the recipient of the money and the evident diversity of opinion which would prevail in discussing this matter, it would seem that the arguments given above with reference to the entrustment of the funds to another organization would be equally valid here. It is the unanimous opinion of this committee that a specific sum, not to exceed the annual income of the fund, should be devoted to the encouragement of such specific research as may be brought to the attention of the committee, and as may seem worthy of receiving this money. It is therefore recommended that the sum of $50 be set aside for the current year and that the Society appoint a committee under whose guidance it shall be expended. ADOLPH FEIEL, Henry R. How.ranp, Macnus PFLAuM, Henry B. Warp, Recorder. COMPLETE LIST OF CONTRIBUTORS TO SPENCER-TOLLES FUND spare, FONT. ws wg ies 6 oie sie ae oo alee 0) eteie sin ‘eine 0) sin ala ciele e/aja alaloim sm am an $ 53 93 One ORT A VE 2 e562. Sic svg wie nin oa um mj otniermy Sralmiajaia inte’ slay = ple nln, ooh ainfale asinine 2 50 Perper om NALA C6 2c oie siere cio oases lo/ oe leila le nha sims te) oletetmtehets J) pial olnvata 1 00 receiiies Dien [eos 2 seize sis ny2e dete ae PAP OR Ay ope ecIaD be ere cits occ 5 00 ee ME PROME IEG Fa18 sia ans 2-2 web sis ci eeisio s cinta ott m apuaelel oe oe atain Sia nies 10 00 iayeesle. Prov: BW ..2)2 . 0). 212 so elas ome sie ia)nre eae niria Hie oles sieislalbe aimee nine 3 00 We EreD ER ODER Mri ee: teresa a esi wal igre erste. Sar oaral epeaeteters weielintc’ eieiaheleys eters inlay Reeteyets 2 00 ere Tense PD goo Hae ats ac id ola Wiallw ot sveprete miotn de areiole an aie, ony ie) el Se 5 00 eri, WHORIAS. (6102 oo eo ole nialn i or eisias ore siete ass De nas Betslo= mdtencrea 5 00 Serta ire Mester si 208, ats). atyaticiel aie ajeta.a sisiaia sit ieceiaeoverauade eleteiara enone mala 10 00 UST SS TN ois el TR ices dt Media Sharon wer che miata ths tg @ Se alan fosncata ite hts Bier ohtace 5d 00 EE PEPE 2-2 3/022 cas lanai scene oleh a erapac yspieltsiseilal anise cian 2 50 eee Pre eae By i. Bs ois cio coolant gals eie satelite selcainaateenras fa.5 5 00 a es OAS US 2 5.5/6 Woo: n!Sn alin seesaw meee nye amen Ariane rate eestaastatelads 4s 10 00 SPENT 2s 9) gS UL UPR Beira ats Aen BUA Oca Quciec toot 35 00 ESTA RRTERLOUEC AI WS cere ois feet AGN i state SeLe PRET toler eed epdcy al ceuet org Mol ocr ohchevavek er of alleys 5 00 Meetiepit. Prot DVS: siiais tsetse sieve detonate eaten tats laa ayeaimiaterelal sis! mts 5 00 268 REPORT ON THE SPENCER-TOLLES FUND Kendall) Dt2 ED erste sk eters bee ee aha Ga ae ETT eas ere ere $ 500 Kenyon, Miss Ada Mii. ii scaiidels wis tie bisie os.ais adler eee 5 00 KEFAUSS DIES Wi Cas Aakers ase erecta, Soke ite Ave cch are aaah eee oe cae ae 1 00 Kanttschnitt; Jobin .scnnenise aes cies enie ease eee eres Ene eee 5 00 Pye wiAs, Wire Ac ee cae este e oes Mekgis stands lene ein, Sperarel dh alioie pepe eirace ave eae ans ae 1 00 Lewis; Dra Wile ctece ial ce oe eat e are en ei Tale Sretave eels A tote 10 00 Maddox Driers siete is eis aie Sree ee acne eet eee a 5 il Manton Dro ligne i scr Ne tae ee Bb fs ba he 2 28 a 5 00 MeKaim: dRiev i ilaslettce hs ier cire cach asa ielesote mcholoterescleie Ofete Wich arts eee 20 00 Mellon dG Geng cclecueccieiicihets car es Woe ER a ay BE Se Ou ae etn eee 20 00 Mercer: (Dr: AliGeie schicics leis cis Uses nae Blas Me esieinee ee eee 1 00 Milnor! | Chass Gur es bee wee lcel wees olslnees ile aae epee nee ae 5 00 Mosprove, Drv Osis ee Ss aug one sie SelB ee ie ets « WO 5 00 INE WCONIEE PHUS io s.c.seeis eine ohae FOS mcs a ee RIE a ae EE ee 5 00 Pennock | Bd ward a. iit wc cach cc eles isle on aula eee ieee 5 00 Piletan Magma. 28 ois bow ciashidin nik are eo nies eels aisles ole alee 14 37 Rogers PrOr. We Aes ache oes cise e ee nye we oni pidicla ele alee ele) 0 eee errr 25 00 Shepard, "Os. (Chas o.oo ec ieeiereeis even pale sislcad wale 8 Sele cies eile 5 00 SHiltz §ChassiS orate dere ele tO oie are fe aban areola a ela aleve le fers Teleh cae ae eens 1 00 Serna fps iie sa )5, clo ana ln' aise ot bia copctone oieicd esti eae he eke ae 15 00 SSRELLER PYM cid ic eo ove je B'S binleld wale’ An: o/c ware Wise! eaysia: cele keen ee 10 00 Mayle, Mee. Ce cel 2 ws siete is Wikiets aa Switch a eis a aie wield wis elese clea olka se! 3 00 VOECE MCMC aac eicehe cea ea wieleee Wi ehoiee es dimye lee atSaiote ak larattlta aero tate cnet ape 10 00 Wand, ‘Profi-H Bes vcacc setbe cle elvis stogulere nine bie Selec eateeled eet ae 10 00 Warde ED rR: EL Cee Nabe sieasie mittee ovemies isle ioe els ters Se ek bis os aR 25 00 Whelpliny, (tS. Wie oe serareo om eisigic bres lsiwsierelnis elbis ie reelateldee eer 2 00 Biol. and Micro. Sect. Academy Natural Sciences, Philadelphia...... 25 00 Butfalo Society Natural Sciences, Buffalo ..............0- 000+ ss0e0: 25 00 Cortland’ Science Club; Cortland, NW... 0.6 a2 ie sian eee eee 3 00 Iron City Microscopical Society, Pittsburg, Pa....... 2.2.2.5 ese. ceee 25 00 New Jersey State Microscopical Society, New Brunswick, N. J........ 25 00 Royal Microscopical Society, London, Eng............0..sce+eceeees 25 20 Speneer Lens \Co.,) Buffalo, N.Y «op .n:2 5 oie -cisscraleieie/aie yaya) ole le ereuelotohe ana 25 00 St. Louis Med. and Surg. Society, St. Louis, Mo..............+eeeees 10 00 Troy Scientific Association, Troy, N. VY... 0... ose ness on aie) se lip else 50 00 Total contributions to February 12, 1902 .............. 200 vee $ 596 61 From) sale‘of Proceedings.) ccc rs eic< inicio nr «ine! slolaicle ole erole ioe Cielo 195 01 Interest and dividends (35). 20.0005 203 2% obec tae + Ue ogee ae ee 416 43 Totals ioc. Ce Gee cece eb ee bie eh hier eNO Se ah eee $1,208 05 The above list includes some contributions received since the last meeting. It seemed proper to have all the names to appear in this volume. MacGnus Priraum, Custodian. Tea oe con CLAYPOLE ER ALL ARD W EDW PEA ECE ee NECROLOGY BDWARD WALLER CLAY POLE,: B.A,,: D.Sc. (LOND.) OF PASADENA, CAL. A truly wise man, loved and respected by all who knew him, has passed on to the other life in the great unknown. The world is better and science and scientific literature are richer because Edward Waller Claypole has lived in this part of God’s universe. Born in Ross, Herefordshire, England, in 1835, he was the eldest of six children, four sons and two daughters. He came of a line of educated and liberal-minded scholars. His paternal grand- father, a clergyman, withdrew from the Church of England to become a Baptist minister. His father, also a Baptist minister, educated in the University of Edinburgh, was a classical scholar. His mother, Elizabeth Mary Blunt, a niece of the English diplomat, Sir Waltham Waller, was brought up in the home of the latter. She received an education superior to that usually given to the young women of her day. This heritage of true refinement and culture, of intellectual ability, of love of justice and truth, of inde- pendence of thought, and of moral courage has been handed down to us increased and enriched. He was educated by his father in classics and mathematics, but his first lessons in science came to him from two sisters of his mother. While he was visiting at their home they interested him in collecting and naming the plants and fossils found in that vicin- ity. These first lessons in science were not from text-books but from Mother Nature, and it may be that at this time he learned not only something of botany and geology but also, unconsciously, something of the value of the laboratory method in teaching sci- ence, which method was one source of power in his later years. At the age of fifteen he began to give instruction in his father’s private school, and about two years later left home to teach at Abingdon. 270 EDWARD WALLER CLAYPOLE In 1854 he matriculated from the University of London, the only English university then open to dissenters. This institution, which gave no instruction, but granted degrees on examination, required of all candidates for the baccalaureate degrees a prelim- inary training in one of certain accredited schools. Since circum- stances made it impossible for him to fulfil this requirement, the young scholar, a dissenter, found no university in all England open to him. Ten years later when this barrier was removed, he passed the examinations for the degrees of B.A. and B.Sc. He might, at this time, by taking further examinations, have obtained the degree of D.Sc., but chose to wait until that degree should be granted for original work, and finally took it in 1888. When nineteen years old, he and two of his younger brothers, Alfred and Henry, all of whom were away from home supporting themselves, undertook as a means of recreation the publication of a magazine called “The Home Journal.” The magazine was not intended for the public and was not printed. Edward, the best penman, and editor-in-chief, received and copied the articles writ- ten by the three contributors, and illustrated the publication with maps and drawings which showed marked artistic ability. The titles of some of the articles that he wrote are “The Power of the Age” (the steam engine), illustrated with section drawings of a locomotive, ‘“The Paralellogram of Forces,” “Corals,” “The Life of Demosthenes,” “First Oration Against Philip” (translated from the Greek), “Laws of Refraction,’ ‘Chemical Nomenclature,” “Geological Formation of Niagara.” This last article was illus- trated by three maps, a geographical map of the country in the region of Niagara, a section showing the geological strata, and a bird’s-eye view of the Niagara river and its vicinity from its head at Lake Erie to its mouth at Lake Ontario. It is of interest to note this work because it shows that already there had been laid the foundation for that breadth and depth of knowledge which after- wards made him distinguished. In 1865 he was married to Jane Trotter, of Coleford, England, a woman of rare beauty of character and moral force. Of the three children born to them, his son Arthur, the oldest, was killed in 1875 by falling from a moving railroad train. The twin daughters, Edith and Agnes, who survive him, have by their original work already won a place among the scientific workers in this country. EDWARD WALLER CLAYPOLE 271 A few weeks after the birth of the twins Dr. Claypole lost his beloved wife—a blow coming as it did at so critical a time in his intellectual life was doubly felt. He was appointed tutor of classics and mathematics in Stokes- Croft College, Bristol, England, in 1867, where he remained for five years. Then there came a crucial test which proved him to be a hero. One of the highest types of heroism known to the world has been shown by those men who, for the love of truth and their unswerving loyalty to their convictions, have suffered persecution. Such a man was Socrates. Such a man was Dr. Claypole. The church at that time branded Darwin’s theory of evolution as heresy. Dr. Claypole saw that it was truth. And because he was firm in his determination to teach the truth he was compelled to resign his position at Stokes-Croft College. How bitter was the cup he drank none of us can realize. He was already sore at heart over the recent loss of his wife, and this step took away the means of support for his three motherless children. But the clouds seemed about to scatter, for he was appointed professor of mathe- matics and natural science in the University College at Aberys- twyth, Wales. At the last moment religious persecution again met him, and he was not permitted to begin work at this institution. So in October of 1872, leaving his little children in care of his parents and sister, he came to this land of freedom in thought and speech, but here, too, he found the same intolerant spirit. Finally, after a year of struggle and patient waiting, he was, through the friendship of Edward Everett Hale, appointed to the professor- ship of natural science at Antioch College, Yellow Springs, Ohio. In 1879 he married Katherine Benedicta Trotter, of Montreal, Can- ada, a second cousin of his first wife, and a woman well fitted to be the companion and helpmate of such a man. He remained at Antioch College until 1881 when, on account of financial diff- culties, its doors were temporarily closed. In the fall of that year he was appointed paleontologist to the Second Geological Sur- vey of Pennsylvania, and spent the next two years in the field. After the close of this engagement in 1883, he accepted the newly established chair of natural sciences in Buchtel College, Akron, Ohio, filling it for fifteen years. Then, on account of his wife’s failing health, he resigned and moved to southern California, taking the professorship in geology and biclogy at Throop Polytechnic 272 EDWARD WALLER CLAYPOLE Institute, Pasadena, a position held until August, 1901, when his work in this world was completed. Dr. Claypole possessed a most happy combination of strong char- acteristics which fitted him to attain eminence both as a teacher and as a scientist. He loved and sought for the truth and would make any sacrifice to uphold it. He had a judicial mind which, having collected all possible evidence, sifted it, weighed it carefully, and considered it from every side before reaching a conclusion, and he had that patience and perseverence which Nature demands from those who successfully interpret her records. As a teacher he was one of the pioneers in using the laboratory method. He made no mental paupers by always “giving men truth instead of training them to search for it.” His students were trained to see, to observe accurately, to record their observations carefully by descriptions and by drawings, to think and reason about them. They were taught the unity of nature and natural law. Their minds were not stored with isolated truths, but the broad relations and general bearing of every truth were made plain to them. Yet his greatest power as a teacher lay in the influence of his character and example upon his pupils. They caught his love for Nature and her truths, his accurate methods of work, his pre- cision of thought, his indomitable perseverence, his inexhaustible patience. Daily contact with such a man was an inspiration even to the dullest and most indifferent of students. They learned to love their work and to love the man who could awaken in them so deep and lasting a desire for knowledge. Many are the tributes paid by those who knew him as a teacher. One of his former students, now a professor in a large eastern university, said to me, “Dr. Claypole is the best teacher that I ever had.” Prof. George M. Richardson, of Leland Stanford Jr. University, a student of Dr. Claypole’s for one year at Antioch College, said, “That one year brought about a complete change in my attitude toward education, a complete change in my ideas as to what education meant, and Professor Claypole was alone responsible for it. I have always felt that he marked out for me my life work. I have never known an- other whose every trait so universally called forth love and admira- tion.” Who can tell how many lives he has inspired with a love of knowledge and a desire to attain to higher ideals of manhood or womanhood ! EDWARD WALLER CLAYPOLE 273 Of the sciences, geology was the most attractive to him, and he is best known for his valuable additions to knowledge in this field. But his many contributions to scientific literature in the fields of botany, zoology, and entomology tell us how truly he loved Nature in all her manifestations, and how familiar he was with every branch of natural science. His first report of original work in geology was given in 1871 while editor of the Proceedings of the Bristol (England) Natural- ists’ Society. In that year and the next he read a series of three papers before the society, “On the subsidence of the southwest counties of England during the present era,’ and one paper on “The development of the carboniferous system in the neighborhood of England.” These earliest writings show the same characteristics that made all of his contributions to knowledge so valuable. In them we find that mastery of the English language which gave such charm as well as force and power to all of his writings and dis- courses; that clear, concise, logical thought, that completeness of evidence, that appreciation of the relation and value of facts, and that ability to interpret correctly the records of Nature. Glacial geology was for him a most fruitful field of study. His paper on “Preglacial formation of the beds of the Great Lakes,” published in the Canadian Naturalist in 1877, and the one read at the meeting of the American Association for the Advancement of Science in 1881, entitled “Evidence from the drift of Ohio, Indiana, and Illinois in support of the preglacial origin of the basins of Lakes Erie and Ontario,” were epoch-making. The views presented in these articles were vigorously combated by some of the other fore- most geologists. But his facts were indisputable and his argu- ments invincible, so that to-day his conclusions reached at that time stand as geological truths. Another plenteous harvest came from his work on the Second Geological Survey of Pennsylvania. Besides the two volumes of the Survey Reports, as a direct result of this period of work, he presented to various scientific associations or published twenty- eight valuable papers. It was on this survey that he found the fossil remains of the hitherto undiscovered genus of ancient fish Palaeaspis. After much patient labor he classified this fossil and proved beyond any question that these were the “oldest indisput- able vertebrate animals the world has yet seen.” To him also be- 274 EDWARD WALLER CLAYPOLE longs the honor of describing Glyptodendron, the oldest of the fossil plants. Another of his rich gifts to American geology and paleontol- ogy was his work on the Devonian Fishes of Ohio. To Dr. Claypole the microscope was a means for many excur- sions beyond the pale of the known. He valued it not only as an aid to scientific research but equally as a means of general educa- tion and as a revealer of nature’s beauties and mysteries to all who would patiently look and search. He took an active part in the meetings of the American Microscopical Society, attending them whenever possible, contributing to its proceedings, and serving as its President in 1897. His address on this occasion on ‘“Micro- . scopical Light in Geological Darkness” was an interesting and brilliant exposition of the secrets revealed by the microscopic study of rocks, secrets which give clues to the geological events in the early history of the world. His wide interest in natural science was also shown by the active part taken in the many scientific organizations of which he was a member. He was one of the founders and the first president of the Ohio Academy of Sciences, a fellow of the Geological Societies of Edinburgh and London, vice-president of section E at the meet- ing of the American Association for the Advancement of Science in 1897, one of the original fellows of the Geological Society of America, and one of the founders and editors of the American Geologists. In the discussions of papers at the meetings of scientific associa- tions he was a power and a promoter of peace when individual animosities began to appear. Often when a discussion had become personal and bitter has he carried it back to the high plane of scientific debate. His was a life of simplicity, purity, and nobility, “full of un- selfish service to others. To have come close to his great nature was a mental and a moral inspiration, and to have known him thus was to love him always.” He belongs to that group of men worthy of the tribute paid to Darwin: “His was a gentle, patient, reverent spirit, and by his life has not only science but our concep- tion of Christianity been advanced and ennobled.” Rogert Orton Moopy. PROCEEDINGS The American Microscopical Society MINUTES OF THE ANNUAL MEETING HELD IN DENVER, COLORADO, AUGUST 29, 20, AND 31, 1901 The twenty-fourth annual meeting of the Society was called to order by President Eigenmann in room 3 of the High School, Denver, Colorado, at 2:00 p.m., Thursday, August 29, 1901. The re- port of the Treasurer was read and referred to an auditing commit- tee consisting of Drs. Bleile and Weber, and it was on motion resolved that the financial year should be closed the Ist of October. The report of the Custodian was read, showing in tabular form the contributions to the Spencer-Tolles Fund during the past year and during the previous period of its existence, the total amount of money received by interest and from other sources, and giving a statement with reference to the investment of the fund. The recom- mendations of the Treasurer were fully approved in a signed state- ment by C. C. Mellor, past Treasurer of the Society, who had examined carefully the character of the present investment. The matter of investment was referred to the Executive Committee with power, and the report itself was referred to an auditing committee consisting of Dr. Schoeney and Professor Beardsley. The report of the special committee on the Spencer-Tolles Fund was read and referred to the Executive Committee with the recom- mendation that the control of the fund be left in the hands of a standing committee to be known as the Spencer-Tolles Fund Com- mittee, but that the apportionment of the fund be charged to the Executive Committee. A new by-law regulating these features was offered as follows: 27 PROCEEDINGS OF THE By-Law No, 1X.—There shall be a standing committee known as the Spencer-Tolles Fund Committee to take general charge of the fund and to recommend annually what part of the income shall be expended for the encouragement of research, but the apportion- ment 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 appor- tioned, under the conditions laid down by the Society for its use. The Custodian shall be an ex-officio member of this committee. The by-law was referred to the Executive Committee. The following by-law was also offered and referred: By-Law No. X.—The Executive Committee shall have the power annually to appoint two members to represent the Society on the Council of the American Association for the Advancement of Sci- ence, in accordance with the regulations of the latter organization. The following amendments to the constitution were offered and laid over one year under the rules: To amend art. III by inserting ‘“‘a custodian,” making lines 5 and 6 read in conformity to this change, “together with a Secretary, a Treasurer, and a Custodian, who shall be elected,” etc. To amend art. IV by inserting in place of “of the Treasurer to act as custodian of” the clause “‘of the Custodian to take charge of,” etc. To amend art. V by adding ‘“‘who still retain membership in this Society.” To amend art. VII by adding “But any person duly elected may, upon payment of $50 at one time, become a life member, entitled to all the privileges of membership, but exempt from further dues and REeSi bd SECOND SESSION At 8:00 p.m. the Society convened in the audience room of the Denver High School for the annual address and reception given under the auspices of the Colorado Microscopical Society. The pro- gram was opened by a piano solo, after which the President of the Colorado Microscopical Society, Dr. A. M. Holmes of Denver, ex- tended a cordial welcome to the national organization. The response was spoken in fitting terms by the retiring President of the Society, Dr. A. M. Bleile. After another musical number the President’s ad- dress was read by Prof. C. H. Eigenmann on “‘The solution of the AMERICAN MICROSCOPICAL SOCIETY 277 eel problem.” It was illustrated by charts and lantern slides, and proved of great interest. At its close an informal reception was tendered the Society and its guests by the Colorado Microscopical Society at which members and guests spent some time in social en- joyment. The occasion was voted one of the most successful which the Society has been privileged to enjoy. THIRD SESSION On Friday, August 30, the Society was called to order at 9:00 A.M. in the Denver High School, Dr. A. M. Bleile being designated temporary presiding officer and Prof. A. E. Beardsley secretary pro tem. during the necessary absence of the regular officers. The by-laws IX and-X were returned from the Executive Committee with a favorable recommendation, and on motion both were duly adopted. The committee then announced as members of the Council of the A. A. A. S. for 1901 Mr. J. C. Smith and Dr. A. M. Holmes. An amendment to by-law No. III was offered as follows: To add “But if not decided, the Secretary shall, unless otherwise ordered by the Executive Committee, print the same number as for the preceding year.” This by-law, being recommended by the Executive Committee, was unanimously adopted. The report of the Committee on the Spencer-Tolles Fund was returned from the Executive Committee and the recommendations adopted as read. The past committee was continued as the Spencer- Tolles Fund Committee, as provided for in by-law IX, just adopted. The thanks of the Society were formally voted to each of the many generous contributors toward this fund, a list of whom is given on page 267, and the Secretary was ordered to send a copy of the com- mittee’s report and of this action to each of them. A nominating committee was appointed, consisting of Dr. A. M. Bleile, Mrs. Cornelia S. Mills, Prof. M. J. Elrod, Dr. L. Schoeney, and Prof. D. Bodine. In view of the considerable number of authors absent, it was resolved that papers be read by title unless the author were present in person. The following papers were then considered: The early morphogenesis and histogenesis of the liver in the pig: D. C. Hilton, Chicago, Ill. 18 278 PROCEEDINGS OF THE The histology of the stigmata and stomata in the peritoneum: A. E. Hertzler, Halstead, Kan. A rearrangement of the families and genera of the Conjugatae: C. E. Bessey, Lincoln, Neb. | A new species of Crenothrix (C. manganifera): D. D. Jackson, New York City. The amount of dissolved oxygen and carbonic acid in natural waters and the effect of these gases on the occurrence of the micro- organisms: G. C. Whipple and H. N. Parker, New York City. Notes on Colorado Protozoa, with description of new species: A. E. Beardsley, Greeley, Col. Notes on Colorado Entomostraca: A. E. Beardsley, Greeley, Col. A review of the American species of Cochleophorus and Curvipes: R. H. Wolcott, Lincoln, Neb. An apparently new hydra from Montana: M. J. Elrod, Missoula, Mont. Some histological features of Echinorhynchi (illustrated) : H. W. Graybill, Lincoln, Neb., was read in brief by H. B. Ward, and a specimen showing the giant nuclei was demonstrated under the microscope. The debt of American microscopy to Spencer and Tolles: W. C. Krauss, Buffalo, N. Y. Some new points on avian cestodes: B. H. Ransom, Bancroft, Neb. A contribution to the subterranean fauna of Texas: C. J. Ulrich, Duluth, Minn. Mounting soft tissues for microscopical examination: M. A. D. Jones, New York City. Modification of some standard laboratory apparatus: S. H. Gage, Ithaca INE? Laboratory photographic apparatus: S. H. Gage, Ithaca, N. Y. The plankton of Lake Maxinkuckee, Ind.: Chancey Juday. Studies on the genus Cittotaenia: R. A. Lyman, Omaha, Neb. A letter was read from Dr. William Trelease, Director of the Missouri Botanical Garden, extending to the Society, on behalf of the local scientific organizations and the Louisiana Purchase Exposi- tion Company, an invitation to hold its 1903 meeting in the city of St. Louis. The letter was referred to the Executive Committee and the question of date and place of the next meeting was also similarly referred. AMERICAN MICROSCOPICAL SOCIETY 279 It was voted to accept the invitation to join the excursion to Boulder, Col., tendered by the University of Colorado for the after- noon, and to take the trip to Colorado Springs, Manitou, and the Garden of the Gods, tendered by Colorado College and the Chamber of Commerce of Colorado Springs on Saturday. It was also voted to hold the final session of the Society on the train en route to Colo- tado Springs, Saturday morning. The Society then adjourned. FOURTH SESSION The Society was called to order by the President at 9:30 A.M. August 31, while en route to Colorado Springs. The Executive Committee recommended that the Secretary be authorized to send some of the back volumes on hand to a selected list of foreign societies, and to add them to the list of organizations to which the Proceedings are regularly distributed. It was further recommended that, in the opinion of the committee, Proceedings should not be dis- tributed gratis in this country, but that in exceptional cases prom- inent foreign societies might receive the Transactions regularly. The report of the committee was adopted, and the Transactions ordered sent to the organizations as designated by the Executive Committee. The Nominating Committee reported as follows: FOR ONE YEAR President, Dr. Charles E. Bessey, University of Nebraska, Lin- coln, Neb. First Vice-President, Dr. E. A. Birge, University of Wisconsin, Madison, Wis. Second Vice-President, Mr. John Aspinwall, New York City. Elective Members of the Executive Committee: Dr. A. M. Holmes, Denver, Col.; Dr. V. A. Latham, Chicago, Ill.; Mr. G. C. Whipple, New York City. FOR THREE YEARS Secretary, Dr. Henry B. Ward, University of Nebraska, Lincoln, Neb. Treasurer, Mr. J. C. Smith, New Orleans, La. Custodian, Mr. Magnus Pflaum, Pittsburg, Pa. The Society appointed Professor S. H. Gage and Mr. Magnus 280 PROCEEDINGS OF THE Pflaum a committee to draw up suitable resolutions of respect for Ex-President E. W. Claypole, expressing its appreciation of the services rendered the organization by Professor Claypole as well as the great loss experienced in his death. [The committee has prepared the following testimonial which, in accord with the instructions of the Society, is spread upon its rec- ords as a token of its indebtedness to one who did so much for its welfare as an organization and for individual members in personal assistance and encouragement. | In the death of Professor Edward W. Claypole science lost a most earnest devotee and this Society a highly esteemed member. He was a conscientious student and his knowledge was profound. His mind was wholly given to his profession, and, as a true scien- tist, he constantly learned and broadened his mind to be better able to impart knowledge. And though the cold facts of science engaged all his faculties to the exclusion almost of the trivialities of common life, yet his heart grew apace, and few men enjoyed a happier fam- ily life in which he gave and received blessings. As a member of our Society his presence at meetings was always a delight. Kind, genial, modest, sincere, he was the center of ad- miring friends, and, with his family, almost an object of envy and an inspiration to others. Of such men the world has too few; he will long be remembered. On behalf of the American Microscopical Society, Macnus PFLauM, Simon Henry GAGE, Committee. The Secretary was instructed to insist on the completion of papers promptly and issue the volume at an earlier date than heretofore. The Society adopted unanimously a vote of thanks to the Colo- rado Microscopical Society and its officers for hospitality extended to the American Microscopical Society, to the musicians who assisted at the reception on Thursday evening, to the many citizens of Colo- rado who officially and personally had contributed so much to make the meeting a success, and especially to those who had aided in planning and carrying out the excursions tendered the organization. On motion the Society then adjourned. Henry B. Warp, Secretary. TREASURER’S REPORT 281 TREASURER’ 5 REPORT FROM SEPTEMBER 20, 1900, TO OCTOBER 15, 1901 DR. aeretenipersiip dies, 1899) oo... ceo kis cas ons inae eae euieciele 3 400 ameersiser ship Hes) TOO) 6.5/0 /.)4'<, << cloie olaie aia @ Hale nee euiaoles 25 00 Ropitenipersuip dies; VOOM eo). .er0/cssis cis se So etale olen nelle: 276 00 SERMMESEICES HIT GMCS, LOUD. i. 0) o.n/c mc aye's abcieis eed oisre a dinero 48 00 —— $ 353 00 opactiiission, feess 1OQU ios... o)s ieee bieeess ciemtwe oul oeteler erate $ 18 00 Mom Ndmission fees, OOD oc cc:06 co.6 s05.cl6 06ers cisreiael Rote vereine 27 00 ; —_— 45 00 SME AT SCHED EHS HEV Ol oy) SON os: 5) o, 0, 75s dval veers rare als ce one alorsrotaes $ 28 00 MAES OSCETDEES 70 V.Ol ey KONIG ooo cio, re a1aibre ~ird «ue eis ctor oretsiacrerets 33 00 —— 61 00 BAP ASIDES ESE AMON. ROR cS aia o: o o'n.e) 0! eicin/k e/e's o/sh's dials) ol a'e & teal $ 200 SEA EMCTLISITIET | VOL, ROE 5 aie oie nen cic aelanars se ge%aaansadels 76 00 —_— 78 00 NMR MERE ANT Ee ete NS 6)y nal o/c) 2y'asc)s gj siaiaia-s «'e "ei sil oidj-0'a' 9s, chelace/ oharera 25 00 pepe ttee Cie PECASHEET 3 5 «: hdee seme sehen $ 388 11 ANNUAL GROWTH Year. Increase Total TSSD se cetcicieisie wcine: cate w wickinte ru Link dawinl va aclveeupae beet Ree ae $ 60 20 TSBG sees Sree a mree - MaRS che res Sema ant Naheid fade Ch $ 25 00 85 20 NESTA Peis nal fate totam focheretaratans ys are els ys atuiak spel nial one caale) ate tae ere 10 00 95 20 PBBB et ioce Heme iak oats end chs Ae lS 1h Ars Nd shuns lcgara te erage era 52 66 147 86 DOSS ete alelen isla tint tate em inije chs wuts is lok aul gia dole molto eet mate 76 00 223 86 FOE Ursa a ec etait n says lie wel eiud dts ee eee Aas es at 30 00 253 86 G host 1 Sahel Mavapedel cg Miah adet (ot ce Se SANS al On ane NR oe ALE Nag YA 39 02 292 88 BBO A eee ee Aten Ae LAE ae mieuiceie ore Moe Perc tcnate iia ele cs BO 19 12 312 00 TBS isa eae kh Mare see a eee ee ee oe at don Beye Os 18 06 330 06 DBDs in sia tutce ts PRT see espa ech nl wv avin Gres aber ai pere fom Bestest spolshnyetel ORNS 19 32 349 38 UE BED aes so iets) tebe eel a TRIB SC ae sh oles east ec tat s eae 22 89 372 27 PEDO Se eateries tA stshe as area iets EPL it das sicae'e ote aol careless Memmi 50 77 423 04 DBOT ois siceis peo emrt eed aaa sll ese tel sa iat he bacco sous 45 99 469 03 BS OB isd hu aa ates lelenis pie oie tote alate chee Uae ae te ee 86 43 555 46 ABD. os 5a Berea ee stentamtans fa: sheptntktet uielet claves 2 Atoteinia ie Cee 97 90 653 36 RDO a's Serel haieter a8 eceue Sikes nla c/a eee intaioielal le & osiie te tete Coe aaa 102 65 756 OL MOOT ss. ree po erste vera lare laren Cre wiatets leis niente, Rime ein Nt eine eae eae ae 388 11 1,144 12 MaAGNus PFLAUM, Custodian. DENVER, CoLo., August 30, 1901. The Auditing Committee appointed to audit the report of the Custodian of the Society report that we have carefully examined the records of the Custo- dian, Magnus Pflaum, together with the records and vouchers thereunto per- taining, and that we find that the said report of the Custodian is in complete accord with the records and vouchers above mentioned. A. E. BEARDSLEY, L. SCHONEY, Auditing Comittee. CONSTITUTION ARTICLE I 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 and Treasurer, who shall be elected for three years and be eligible for re-election. 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 Treasurer to act as custodian of the property of the Society, and of the Secretary to edit and publish the Proceedings 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. 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. 284 CONSTITUTION AND BY-LAWS ARTICLE VII The initiation fee shall be $3, and the dues shall be $2 annually, payable in advance. ArrtIcLe 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 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. II The Secretary shall edit and publish the papers accepted with the necessary illustrations. Ill The number of copies of Proceedings of any meeting shall be de- cided at that meeting. But if not decided, the Secretary shall, unless otherwise ordered by the Executive Committee, print the same num- ber as for the preceding year. 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 Proceedings shall not be sent to any member whose dues are unpaid. CONSTITUTION AND BY-LAWS 285 V The election of officers shall be held on the morning of the last day of the annual meeting. Their term 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. 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. 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. VIll Members of this Society shall have the privilege of enroling 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). 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 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. 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, August 30, 1901. 3 Orkney OV he orl are ate aut h¢, ALINE AN nf aa ha hte pac en! i AGH wu Abb Pe EL Cts, Pane a ey hiae ny ' i we eas / We dl | ryaya ie War NM SUH PEAR aon ‘ ey te ry | t " ele ic WANN aM HER LIST OF 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 1901 For addresses see regular list. CHARLES, Jos. WM. LYMAN, RuFus A., A.M. DISBROW, WILLIAM S., M.D. MARSH, Jas. P., M.D. FOore, J. S., M.D. MILEs, MRs. C. S. GALLOWAY, PROF. T. W. VE ENG Ce ie) GRAYBILL, H. W., B.Sc. MOCKETT, J. H., SR. HALL, VICTOR S. SARCAR, HEM CHUNDRA, M.B. Hi_Ton, Davin C., A.B. STAUFFER, REv. T. F. HERZOG, MAXIMILIAN, M.D. STEBBINS, J. H., M.D., PH.D. KINLEY, Jos. B., M.D. ULRICH, PROF. CARL J. ABER DEIN | ROBERT MLD) 82.5 v5, edelecisajae.cce 327 James St., Syracuse, N. Y. AMIGTERE LC EVAR ICES) Nis, 2925. « 2c-c.cis)s1e/e 8) selec sie.e.elsietsveisteleierseleeets Rochester, Minn. ALLEGER, WALTER W., M.D., ’94........ 949 T St., N. W., Washington, D. C. SPUN CAMEE SJ OEUN A WGC Al: OO) ste.) che: die/nierc(oiersielsla als, alstels Mat s\aitretors Newburg, N. Y. Arwoon, E.S., °79...... sedtaavodeone Highlands P. O., Monmouth Co., N. J. INN OIOUW AEM OH 7 1Olsis.s's)s/2ie\vi pisces nyeleus exer’ 16 Seneca Parkway, Rochester, N. Y. PUES ORGAN, W..) MOD), BT. sac ce sieice.n cine sepa sialels Upper Montclair, N. J. BARKER, ALBERT S., ’97....Twenty-fourth and Locust Sts., Philadelphia, Pa. BARNSFATHER, JAMES, M.D.,’91..... Sixth Ave. and Walnut St., Dayton, Ky. BARTLETT, CHARLES JOSEPH, M.D., ’96...... 150 York St., New Haven, Conn. BAUSCH EDWARDS °(8)sic:c.0/, (?96.5 12 :). 12 6c /e(s/cys 4 siniels 1833 Mariposa St., Fresno, Cal. ULRICH, /CARE Me PBS OWE. castes siesre's Central High School, Duluth, Minn. VANDERPOEL, FRANK, M.E., Ph.D., ’87........ 153 Center St., Orange, N. J. VEEDER Ss IMCVACMMCT (38 ler cayeieroala are, opuel steers 12 Queen Sit., Lyons, N. Y. VorcE, C. M., Esg., F.R.M.S., ’98..... 209 Cuyahoga Bldg., Cleveland, Ohio VREDENBURGH, Bi Hi, 7840. ci2)0). 36200 3 60 Plymouth Ave., Rochester, N. Y. WAL MSE EVE Wi rEls ilGesiecetene ciataleiaiereyestaterserohs 4248 Pine St., Philadelphia, Pa. WARD, HENRY B., A.M., Ph.D., ’87.... University of Nebraska, Lincoln, Neb. WEBER, PRoF. HENRY A., Ph.D., ’86........ 1342 Forsyth Ave., Columbus, O. WREKS, JOHN ROGKWEETA 90 oon). cciec «oles siarcelsts Weather Bureau, Macon, Ga. WEIGH IWAN, | CHAS. es BGr eeu isaieici eile 5859 Michigan Ave., Chicago, Ill. WHECH, 'GEOWO} MED Ol ome nee creme icis Box 416, Fergus Falls, Minn. WELLINGTON, CHARLES, 999.......000c000s 403 Pringle Ave., Jackson, Mich. WENDE, ERNEST, M.D., F.R.M.S., ’91..... 471 Delaware Ave., Buffalo, N. Y. WHEELER, B,J.) PRD: (000 aye aniaeace 79 Chapel St., Albany, N. Y. WHELPLEY, H. M., MED., Ph.G., F.R.M.S., 90, 2342 Albion Place, St. Louis, Mo. LIST OF MEMBERS 293 MirmpPE, (GC. 709)... fo... 5s Director Mt. Prospect Laboratory, Flatbush Ave. and E. Parkway, Brooklyn, N. Y. WHITLEY, JAMES D., M.D., F.R.M.S., ’85...405 So. Main St., Petersburg, Ill. AGU ee MCAT TING S50 SO) o/s occ else e's sia sess oe 21 Walnut St., New Britain, Conn. WILLSON, LEONIDAS A., Eso., ’85......... 112 Public Square, Cleveland, Ohio WOLCOTT, ROBERT HENRY, A.M., M.D., ’98, University of Nebraska, Lincoln, Neb. Younc, Aucustus A., M.D., ’92..22 E. Miller St., Newark, Wayne Co., N. Y. SOLE ap lee elds o/ojc lec acc di sicie's siorstete'e led a v alee « High School, Evansviile, Ind. ZENTMAYER, FRANK, ’91 ......... _....209 S. Eleventh St., Philadelphia, Pa. HONORARY MEMBERS CRISP, FRANK, LL.B., B.A., F.R.M.S., 5 Landsdowne Road, Notting Hill, London, England DALLINGER, REv. W. H., F.R.S., F.R.M.S., Ingleside, Lee, S. E., London, England EONSON, Ce -r.,-A.M., L.D.,. F.R.M.S., Hillside, Clarence Road, Shanklin, Isle of Wight, England Mappox, R. L., M.D., Hon. F.R.M.S. Greenbank, 45 Belmont Road, Portswood, Southampton, England SNERBH ANENE-TONI IY. WEDD. 5 cccaccis sine’ 21 Federal St., New London, Conn. WARD R EATLSTED, Aj M., M:D., E.R.M.S....0<.; 53 Fourth St., Troy, N. 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SO a “ “a, - = - ~~ n ~ us a ede: Se epee: ah nyo version wend Menta oe Rr ORE hy > tan haber tess ne rer er