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‘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
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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.
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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
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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. <A carefully trained collector would ordinarily do effective work, but the results obtained on his samples must be checked by work in the field if a com- prehensive idea of the growths and the conditions which
FIELD-WORK ON SURFACE WATER-SUPPLIES 21
make for and against their development is to be obtained. The field-work must not be conducted in a haphazard way, but the general principles which influence plankton growths must be kept in mind.
As the name implies, the plankton is made up of floating growths, and many of them float near the surface of the water, while others live at some depth beneath it. But all are in- fluenced by currents and winds. There is a steady current from the influent to the foot of the reservoir, so that there will be a tendency for all forms to be swept toward it. Then the wind will blow the growths hither and thither over the pond, now toward one shore, now toward another. Not only does the wind waft them from place to place, but distributes them also in the vertical through which it acts. So those forms which usually grow in the upper five feet and those which float conspicuously on the surface will be mixed with those which develop at a depth.
When the wind subsides they again take their normal posi- tion in this vertical. It is easy to see how all this may influ- ence the samples. In the first place, organisms may be blown towards or entirely away from the usual collecting points. So easy is it to be deceived by this occurrence that it is not a bad plan to have the direction of the wind noted on the sample tag. If the samples are taken soon after a high wind they will give an entirely false idea of the vertical distribu- tion of the organisms. Moreover, a small growth may be carried from its place, and collect about the intake. A bottle received from the collector at this time will create uneasiness which will be dispelled by a visit to the works where the true size of the growth is revealed, together with the fact that it will be dissipated again by the wind, leaving the water as good as ever. !
Matters do not always work out as satisfactorily as this. These microscopic organisms commonly develop in bays and nooks where there is a shallow flowage, and in places where the cleansing of the reservoir has not been thorough. It hap- pens at times that they are blown from these places out into the reservoir,and there increase enormously, causing the water to become very offensive to taste and smell. If the biologist finds a growth of this kind on his hands his procedure will be determined by the results of his field-work. If they show that the growth is confined to the surface, water may still
22 HORATIO N. PARKER
be drawn from the mid-depth and bottom. This state of affairs many times exists in the case of growths of cyanophy- ceae and of some infusoria. But if it has been learned that the whole reservoir, from surface to bottom, from foot to head, is infested there is nothing to do but shut off the supply, and wait until the vigor of the growth is abated before using it again. Now this is a vital point. By putting the works in the care of a competent biologist, and constructing them so that water may be drawn from the surface, mid-depth and bot- tom, aS occasion may require, it is perfectly feasible to deliver at all times good tasting water to the users. Even in the cases where there is only one source of supply much may be done in this direction, and where there are many sources to sup- plement each other the water should always be good. Our municipalities have not yet learned this lesson, but they must be made to as a matter of decency and economy. The micro- scopic organisms, besides developing in the shallows and bays of a reservoir, may be brought into it by the influent streams. Ordinarily they do not carry many organisms, and those that they do carry are quite harmless forms, such as Antho- physa, a few chlorophytes, and diatoms. But if the streams expand to broad quiet pools in some places, or if they rise in Swamps which contain ponds, they may bring from thence many organisms which will work harm in the reservoirs below. These little swamp ponds and pools in the streams are ideal places for certain infusoria and cyanophyceae to breed, and they may reach a large development there, being retained in these localities as long as the weather is dry, and the streams are low. A heavy rain, however, washes them into the reser- voirs where they are likely to grow, and do much injury. I believe many storage reservoirs which have been stripped at great expense have been spoiled by neglecting to eliminate the ponds from swamps and the pockets from the streams.
It is in this connection that we should consider the effect of the abandoned mill-ponds on some of the feeders of the chain of reservoirs. In summer time the microscopic organ- isms are bound to develop abundantly in them, and a long rain will make them overflow, and deliver their noxious growths into the stream; once there, the journey to the reser- voirs is a short one. These ponds are always a great cause of worriment to the biologist. Samples should be taken from them often enough to determine their predominant organisms, and the grade of their water surface should be read from time
FIELD-WORK ON SURFACE WATER-SUPPLIDS 23
to time, that they may not overflow unexpectedly, and so start a considerable growth, which may remain for a time at the head of the reservoir, unobserved by the biologist.
The seasons of overturns of the ponds are periods of great activity in the field. As this phenomenon is dependent on the temperature of the water many temperature readings ought to be made before and after it occurs. The overturn is suc- ceeded by a rapid multiplication of diatoms and other forms, so that many samples have to be taken for microscopical ex- amination. It is not till the material brought up from the bottom has been oxidized, and has settled out, and the microscopic organisms have in consequence somewhat de- creased that there can be any let up in this work.
Some water works are built so that one reservoir can be filled from another, and not infrequently it is necessary to do this. Here is another chance for field-work, for the empty basin should never be filled with water of questionable quality. When such a work is to be undertaken, the biologist should invariably visit the works in person and assure himself that there is no marked turbidity, that the color is low, and that there are few organisms. It would be the height of folly, for instance, to fill an empty reservoir in mid-summer when so many organisms of all classes are present in the water. With the advent of cold weather many forms die so that the actual number of organisms is less, but of vastly more importance is the fact that many of the spores, which in warm weather are somewhat generally distributed in the water, sink to the bottom, and there remain till the spring overturn brings them to the surface and sunlight again. The significance of this is that we have a choice; we may fill our reservoirs with water which is comparatively free from organisms, and is likely to remain so, or we may fill them with water holding a multitude of forms which in time will die, leaving seed-like bodies, which will inevitably germinate as soon as conditions favorable to their development recur. In the light of these facts, it seems as though we ought always to act cautiously and wisely in the matter of running the water from one place to another in the system. But at times an apparent necessity tempts us to take chances which prudence would forbid our accepting. If we yield to these promptings, it should be with a full realization of the fact that it is easy to give the microscopic organisms a foothold in our reservoirs, but that it is not so certain that we are ever entirely rid of them again.
24 FIELD-WORK ON SURFACE WATER-SUPPLIES
Elsewhere, I have spoken of nitrates in a manner which in- dicates that they are deleterious to a supply. It is only right to qualify this, for they are not injurious to health, but they are a detriment in that they serve as a food for the algae. Now the nitrates of a watershed increase with population; consequently, when it becomes very dense we may expect much trouble from these alone. In ground water supplies we ordi- narily find high nitrates, but they are not harmful so long as the sunlight is excluded. Under its influence, however, the chlorophyllous plants produce extremely abundantly. Indeed so marked is this tendency that reservoirs for waters of this class are usually covered.
Many examples of the efficiency of field-work might be quoted to emphasize its value. But as we have treated the subject in a general way it does not seem fitting to do this. It will be sufficient to cite one case where its results were much appreciated by those for whom it was undertaken.
The town of A bought water from the Metropolitan Water Board; at the same time it used some of its own water, and this mixed supply was delivered to the consumers. The A supply was filtered water taken from filter-galleries near the shore of a lake, and this was mixed with the surface water supplied by the Board. The mixing did not take place in an open reservoir but in the mains. For a time all went well, but suddenly the town of A complained that the Metropolitan water was unfit to drink. I at once investigated the matter. Samples were taken from the reservoir which the Metropolitan Water Board used to supply A, from the A taps, from the A filter-galleries, and from the lake near which the filter-galleries were built. The analyses showed that the water from the Metropolitan reservoir was good, and that from the A taps was bad. It followed that the trouble must be with the water supplied by A itself. There were two organisms, Crenothrix and Coelosphaerium, in the A taps water which were not in the Metropolitan water. The water from the filter-galleries contained Crenothrix but not Coelosphaerium. ‘The water from the lake contained great quantities of the latter, and as it was this which was causing the trouble it was important to discover how it got into the supply. The superintendent bethought himself of a pipe which was formerly used to let water direct from the lake to the town mains. A visit to this solved the mystery; the valve which closed the pipe was leaking badly. When it was replaced the trouble stopped.
THE WORK OF MT. PROSPECT LABORATORY OF THE BROOKLYN WATER WORKS
By GEORGE C. WHIPPLE, C. E., BrioLoGist AND DIRECTOR
WITH FOUR PLATES
The practical value of the sciences in our modern civili- zation is strikingly attested by the increase in the number of laboratories connected with various departments of nation, state and municipality. This is emphatically true in the do- main of sanitary science, where the advances in chemistry, microscopy and bacteriology have wrought revolutionary changes. With the knowledge that many diseases are caused by living organisms, and that some of them are transmitted by water, came the need of more careful supervision of public water supplies, which resulted in the establishment of labora- tories devoted to water analysis. The pioneer work of the Massachusetts State Board of Health has been followed by the installation of laboratories in most of our large American cities. In many instances these are operated in connection with the departments of health, but in Boston, Lynn, Louis- ville, Cincinnati, Pittsburg, Albany, Washington, and else- where laboratories have been organized in connection with the departments of water supply, either for the purpose of ex- perimental work or because the character of the water supply was such that proper management depended upon analytical as well as engineering data. Departments of water supply should be justly held responsible for the quality as well as the quantity of water supplied to the public. This involves a con- stant knowledge of the sanitary condition of the water, which can be obtained only by frequent analysis and inspection.
The complicated character of the water supply of Brooklyn made the need of a laboratory apparent to the Department of Water Supply several years ago, but it was not until 1897 that an appropriation for the purpose was obtained. In May of that year the writer was appointed biologist and director of the laboratory, and instructed by Mr. I. M. De Varona,
26 GEORGE C. WHIPPLE
Engineer of Water Supply, to prepare plans for the installation and equipment of a complete chemical and biologi- cal laboratory. Mt. Prospect reservoir, near the entrance to Prospect Park, was selected as the most available site, and the gate house of the reservoir was found to have ample accom- modations. Contracts were let during the summer, and the laboratory was in complete operation on the 1st of October, though regular microscopical examinations of the water were begun early in July.
Mt. Prospect laboratory has a fortunate location. It is con- veniently near the Long Island depot, where the samples from the watershed are received, Ridgewood reservoir, the main dis- tribution reservoir, and the office of the department at the municipal building. Its isolation and elevation make it com- paratively free from noise and dust, while the building is well lighted by large windows, heated by hot water, provided with gas, electricity and telephone. The upper portion of the build- ing contains three rooms, besides the keeper’s office, and a cor- ridor from which visitors may ascend to an observatory on the roof. The three rooms are known as the general laboratory, or preparation room, the biological laboratory, and the chemi- cal laboratory. In the basement are the physical laboratory, the general store room and the furnace room. There is also a sub-basement, suitable for bacteriological work during hot weather.
The general laboratory is devoted to the shipment of bottles and reception of samples, the washing of glassware, the steril- ization of apparatus, the preparation of culture media and to such chemical operations as might charge the air with am- monia and the fumes of strong acids. The room contains a well-ventilated hood; a work table, under which are closets and drawers; a shipping desk; a large sink, with draining boards on the sides; a drying oven; a hot-air sterilizer; a steam sterilizer; an autoclay; an automatic still, and a distilled water tank, lined with block tin and having delivery tubes that ex- tend to the other rooms.
The biological laboratory is devoted to the bacteriological and microscopical examinations of water and to the study of the various organisms found. It also forms the office of the director. It contains a work table; a long work shelf, with three windows in front; three incubators; an ice chest for the storage of culture media; a case for sterilized apparatus;
WORK OF MT. PROSPECT LABORATORY 27 a bookease, with a good working library; a desk; a typewriter, and cabinets for report blanks, biological specimens, ete. Electric bells connect with the different laboratories and with the telephone in the office of the keeper of the reservoir.
The chemical laboratory is the largest of the three rooms. Its atmosphere is kept free from ammonia and from the fumes of strong acids, in order not to vitiate the results of the water analyses there carried on. It contains a table for hold- ing the samples of water that are being analyzed; three work tables; two work shelves, with windows in front; a weighing room, with balance in front of window and with a wide desic- cator shelf and a drying closet; a hood, under which are two steam baths; a battery of twelve stills for ammonia distilla- tions; a still for obtaining redistilled water; an apparatus for gas analysis; a battery of twelve Sedwick-Rafter filters, used in the microscopical examination of water; an apparatus case; a case for chemicals; a Richards pump, and various pieces of specially designed apparatus that facilitate the work of analy- sis. The room also contains a combustion furnace and a Mahler bomb calorimeter for the analysis of coal and the determina- tion of its heating power. A storage room opens from the chemical laboratory, and there is a small dark room under the stairs. The three laboratories have marble-tiled floors, and the work tables and shelves are covered with white tiles throughout. The partitions between the rooms are largely of glass.
The physical laboratory in the basement is not fully com- pleted. At present it contains a crusher, a coal sampler, sieves for sand analysis, and a complete equipment for testing ce- ment. Much of the room is devoted to storage and to shop work.
The laboratory force consists of one biologist and director, one chemist, one assistant chemist and three assistants.
WATER ANALYSES
The routine work consists of the regular examination of samples of water received from all parts of the water-shed and distribution system, i. e., from the driven wells, streams, ponds, aqueducts, reservoirs and service taps. The compli- cated and varied character of the water supply requires the examination of an unusually large number of samples, and it is safe to say that no water supply in this country is examined
28 GEORGE C. WHIPPLE
more thoroughly and minutely than that of Brooklyn. The regular routine includes the bacteriological examination of three samples of water from the Ridgewood pumping stations and from a tap in the city collected daily; the complete physi- cal, chemical and biological examination of nine samples from the distribution system collected weekly; the physical, biologi- eal and partial chemical examination of 24 samples from the supply ponds collected weekly, with complete chemical analy- ses monthly, the complete examination of 19 samples from driven wells collected monthly; and the complete examination of 21 samples from the private water supply companies of Brooklyn and from the water supplies of the Borough of Queens, collected quarterly. In addition to these regular samples many extra samples are taken at various times and places, as occasion requires. During the 23 years that the laboratory has been in operation this schedule has resulted in the analysis of more than 6,000 samples, as follows:
Samples received from July 12, 1897, to April 1, 1900... .6,471
Physical Cxamimations 0.0/5.2 - pes oe oun oe Se 5,025 Complete;chemical ‘analyses... .0. 2 5 ./4)s)...10iys ese ee 2,562 Partial: chemical ‘analyses \iciiai) 60k isaa's sees ene 1,049 Microscopical examinations |. . 4640.60. 2 Ue ee 4,688 Bacteriological examinations, 6.0. 2. we. c/s cseeeneneene 5,230 Tests for (bacillus: coli) communis , i... sc. £).' ee 2,630
The samples of water from the watershed are collected in the forenoon during the early part of each week and sent to the laboratory by express. The average time that elapses between the collection of a sample and the beginning of its analysis is about four hours, but this time varies from ten minutes to eight hours. Samples are collected in large bottles for chemical and microscopical analysis and in small sterilized bottles for bacteriological examination. The large bottles have a capacity of one gallon, are made of heavy, clear, white glass, and have glass “hood” stoppers. They are not sterilized, but are carefully cleaned with chromic acid before leaving the laboratory. Brown paper is tied over the stoppers to prevent contamination from dust, and the bottles are packed in boxes that have separate compartments lined with indented fiber paper and that are provided with tight-fitting covers. The breakage of bottles packed in this way is very small.
The bottles for the bacteria samples hold 2 ozs., and are made of clear, white glass, and have wide mouths with glass
WORK OF MT. PROSPECT LABORATORY 29
stoppers. They are known to the trade as “chemical salt mouths.” These bottles are sterilized each time before use. The stoppers of the bottles are covered with pieces of tin foil, and each bottle is then placed in a screw-capped tin box, just large enough to receive it. The tin boxes are painted to keep them from rusting. The bacteria samples are shipped in port- able ice-boxes. There is an outer box with asbestos packing and a copper lining and an inner copper tray, divided into compartments to hold the tin boxes just mentioned, and be- tween the outer box and the tray is a large space for ice. The box holds sufficient ice to last eight hours in hot weather, and the samples almost invariably are received in good condition.
The samples from the supply ponds are collected at a depth of 1 ft. below the surface. The shallowness of the ponds makes it unnecessary to collect samples at greater depths. The samples from the distribution reservoirs are collected just outside the gate houses, where the flowing water gives a rep- resentative mixture of the water entering or leaving the reser- voirs. Special precautions are taken to avoid contamination in the collection of samples, and to this end special forms of collecting apparatus have been devised.
In the apparatus for collecting the bacteria samples the sterilized bottle is placed in a metal frame attached to the lower end of a small brass tube, and is held in position by spring clips. A small rod extends through the brass tube, and at the lower end is provided with a clutch for grasping the stopper of the bottle. By means of this rod the bottle may be opened and closed under water.
The apparatus used for collecting samples from beneath the surface, when necessary, is as follows: The frame consists of a brass wire attached to a weight with clips for holding the bottle. The frame is supported by a spring joined to a sinking rope. A flexible cord extends from the top of the spring to the stopper of the bottle. The length of this cord and the length and stiffness of the spring are so adjusted that when the apparatus is suspended in the water by the sinking rope the cord will be just a little slack. In this condition it is low- ered to the desired depth. A sudden jerk given to the rope stretches the spring and produces sufficient tension on the cord to pull out the stopper.
The apparatus for collecting bacteria samples from beneath the surface is similar in principle. The bottle is replaced by a
30 GEORGE C. WHIPPLE
sterilized vacuum tube, with end turned outwards and back- wards and drawn to a point. The pull of the cord breaks off the tip of the tube and the pressure of the water causes the tube to fill. The end may be then sealed with an alcohol lamp or closed with a bit of sterilized wax. The frame for holding the tube consists of a short piece of lead pipe, which also serves as a weight.
The temperature of each sample is taken at the time of collection and recorded on a certificate, together with the locality of the sample, the date of collection, the name of the collector, etc. Temperature reading below the surface are obtained with the thermophone.*
When the samples reach the laboratory each is given a serial number and entered in an index book, and throughout all the examinations each sample is known by its number rather than by the name of the locality from which it was | collected.
It would be out of place in this paper to describe in detail all the methods used in the analysis of the samples, but inas- much as methods differ considerably in different laboratories, it seems desirable to give at least an outline of the methods used and to describe such as differ materially from those practiced elsewhere.
PHYSICAL EXAMINATION
The physical examination includes the observation of the temperature of the water, its general appearance, its tur- bidity, its color and its odor. -
Temperature.—The temperature of the sample is observed at the time of the collection, as mentioned above.
Appearance.—The amount of sediment and the turbidity, after standing twelve hours, are estimated by inspection and recorded numerically according to the following scale: 0, none; 1, very slight; 2, slight; 3, distinct; 4, decided.
Turbidity.—The actual turbidity is determined by compari- son of the sample with silica standards of turbidity, as de- scribed by Whipple and Jackson in the Technology Quar- terly for December, 1899, and September, 1900. According to this standard, a turbidity of 100 is equal to that produced by adding 100 mg. of finely divided diatomaceous earth to one liter of water. Comparisons are made in gallon bottles or in
* Henry E. Warren and Geo. C. Whipple, ‘‘The Thermophone,”’ Technology Quarterly, July, 1895.
WORK OF MT. PROSPECT LABORATORY 31
Nessler jars, held toward the light or placed over a series of black lines.
Color.—The color is determined by comparison with the platinum-cobalt standard, described by Hazen in the Ameri- can Chemical Journal, Vol. XIV, p. 300. The comparisons are made in 100 cu. cm. Nessler jars, 1 in. in diameter and 12 ins. long.
Odor.—The “cold odor” is observed after vigorously shak- ing the bottle in which the sample is contained. The “hot odor” is observed by heating about 200 cu. em. of the sample in a beaker, covered with a watch-glass, to a point just short of boiling and applying the nose as soon as the water has sufficiently cooled. The results are expressed according to the following scale of intensity and with the following abbre- viations:
Scale of intensity—0, none; 1, very faint; 2, faint; 8, distinct; 4, decided.
Abbreviations.—v, vegetable; e, earthy; a, aromatic; g, grassy; f, fishy; m, moldy, etc.
CHEMICAL ANALYSIS
The sanitary chemical analysis ordinarily includes the deter- mination of the nitrogen as albuminoid ammonia, free am- monia, nitrites and nitrates; total residue on evaporation, loss on ignition, chlorine, iron and hardness. In addition to these the following determinations are sometimes made: oxygen consumed, alkalinity, incrusting constituents, dis- solved oxygen, carbonic acid, ete.
Form of Expression.—The results of the chemical analysis are expressed in parts per million.
Nitrogen as Albuminoid Ammonia.—The method of Wank- lyn is used, according to the practice of the Massachusetts State Board of Health, described in the two special reports on water supply and sewerage published in 1890. The total albuminoid ammonia is determined on the unfiltered water. The dissolved albuminoid ammonia is determined after filter- ing the sample through filter paper. The suspended albumin- oid ammonia is found by subtracting the dissolved albuminoid ammonia from the total albuminoid ammonia. In the case of ground waters only the total albuminoid ammonia is deter- mined. The form of distilling apparatus is practically the same as that designed by Mr. H. W. Clark and used at the
32 GEORGE C. WHIPPLE
laboratory of the Massachusetts State Board of Health. Per- manent standards are used as described by Jackson in the Technology Quarterly for December, 1900.
Nitrogen as Free Ammonia.—The free ammonia is deter- mined by Wanklyn’s method, referred to under albuminoid ammonia. Five hundred cu. cm. of the sample serves for the determination of both the free and albuminoid ammonia.
Nitrogen as Nitrites.—Warrington’s modification of the Griess method is used. Permanent standards are used.
Nitrogen as Nitrates.—The phenolsulphonic acid method of Grandval and Lajoux is used, but with certain modifications tending to refinement. The quantities of water operated upon vary from 2 to 50 cu. cm., according to the amount of nitrogen present as nitrates. Permanent standards are used instead of preparing fresh standards for every set of compari- sons. Comparisons are thade in 100 cu. cm. Nessler jars.
Residue on Evaporation.—For the determination of the residue on evaporation 200 cu. em. of the sample are evapo- rated to dryness on a water bath in a platinum dish of known weight, dried for half an hour in a steam oven, cooled in a desiccator and weighed. Where it is necessary to determine the amount of suspended matter the residue is determined both before and after filtering the sample through filter paper or through a Pasteur filter, and the difference obtained.
Loss on Jenition.—After the determination of the total resi- due on evaporation the platinum dish is placed in a larger platinum dish that serves as a radiator, ignited for seven minutes at a low red heat, treated with a small amount of distilled water to restore any loss of water of crystallization that may have been driven off by the ignition, evaporated to dryness on the water bath and dried, cooled and weighed as before. The difference of weight before and after ignition gives the loss on ignition. The loss on ignition is not deter- mined for the ground waters or‘for the water of the distribu- tion system, which is a mixture of the surface and ground waters.
Chlorine.—The chlorine is determined by titration with sil- yer nitrate, using potassium chromate as an indicator, accord- ing to Hazen’s modification of Mohr’s method, described in the American Chemical Journal, Vol. XI, p. 409.
Hardness.—The hardness is determined by Clark’s soap method, substantially as described in Sutton’s “Volumetric
WORK OF MT. PROSPECT LABORATORY 33
Analysis,” but with certain modifications in the preparation of the soap solution. No attempt is made to separate the “temporary hardness” from the ‘permanent hardness” by the method of boiling. The information covered by these terms is obtained when required by the determination of the alkalin- ity and the incrusting constituents.
Alkalinity—The alkalinity of a water is a measure of the carbonates and bicarbonates present. It is ordinarily deter- mined by titrating 100 cu. cm. of the sample with N-50 H,SO,, using methyl orange as an indicator; but it is some- times desirable to substitute lacmoid for methyl orange as an indicator, making the titration after heating the sample to the boiling point. Phenacetolin is also used. It has been found that when the true end-points are known and the proper cor- rections are applied the various indicators give practically the same results. These indicators differ in their power of show- ing the presence of sulphate of alumina, and methyl orange should not be used in determining the alkalinity of a water that has been treated with that coagulant.
Incrusting Constituents.—The incrusting constituents are the salts that give to water its “permanent hardness.” The determination is made according to Hehner’s method, as de- scribed by Leffman in his “Examination of Water.” The sum of the alkalinity and incrusting constituents is approximately equal to the hardness as determined by the soap method.
Iron.—The iron is determined from the residue in the platinum dish according to Thompson’s method, as described in Sutton’s “Volumetric Analysis,” but with certain changes in technique that tend to greater accuracy.
Oyxgen Consumed.—The Kubel method is used _ sub- stantially as described in the special reports of the Massachu- setts State Board of Health, above referred to. The period of boiling is five minutes. This determination is seldom made on the regular samples.
Dissolved Oxygen.—Winkler’s method is used according to the modifications of Drown and Hazen, described in the special reports of the Massachusetts State Board of Health, above referred to.
Carbonic Acid.—Pettenkofer’s method is used according to the modifications of Trillich, described in Ohmuller’s “‘Unter- suchung des Wassers,” edition of 1896, when it is desired to determine the free and half-bound carbonic acid. The free
3
34 GEORGE C. WHIPPLE
carbonic acid is determined by titrating with N,,NaOH, using phenolphthalein as an indicator.
MICROSCOPICAL EXAMINATION
The microscopical examination of water determines the number and kind of microscopic organisms present, together with the amount of amorphous matter. The Sedgwick-Rafter method is used, with the modifications described in the author’s “Microscopy of Drinking Water.” The results are expressed in number of standard units of organisms per cubic centimeter.
BACTERIOLOGICAL EXAMINATION
The bacteriological examination consists of the determina- tion of the number of bacteria present in a sample of water and a qualitative test for the presence of bacillus coli com- munis. No general qualitative work is undertaken in connec- tion with the regular routine.
Quantitative Examination.—One cubic centimeter of the sample (diluted 1-10, or 1-100, if necessary) is mixed with 5 cu. cm. of sterilized nutrient gelatine in a ventilated petri dish and allowed to cool on a level surface. When hard the culture is placed in an incubator and kept at a temperature of 20° C. in an atmosphere saturated with moisture for 48 hours, after which the number of developed colonies is counted. It is then returned to the incubator and kept 24 hours longer, after which a second count is made. The 72-hour count is the one reported. All determinations are made in duplicate. The gelatine used as the culture medium is prepared substantially as recommended in the report of the Bacteriological Com- mittee of the American Public Health Association, published in 1898. It is given an acidity of 1.54.
Test for Bacillus Coli Communis.—Smith’s fermentation method is used as the basis of the test, isolation of the colon bacillus according to ordinary qualitative methods being at- tempted only when a positive test is obtained in the fermenta- tion tube. If the amount of gas in the fermentation tube after 48 hours’ incubation at 37° C. is above 30¢ and below 704 of the closed arm, a portion of the sediment is plated on lactose- litmus-agar. If red colonies develop after 12 hours’ incuba- tion transfers are made from them to glucose-gelatine, milk, nitrate solution, sugar-free broth (for indol), and glucose broth
WORK OF MT. PROSPECT LABORATORY 35
in a fermentation tube. If these tests give positive results, the presence of the colon bacillus is considered as proven.
The members of this Society will be naturally most inter- ested in the results of the microscopical examinations. These cannot be described in detail within the compass of this paper, but the following account of some of the more important microscopic organisms will indicate the nature of the prob- lems that are being investigated.
MICROSCOPIC ORGANISMS IN THE BROOKLYN WATER SUPPLY
The troubles of the Brooklyn water supply during the past few years have been occasioned by the growth of odor-produc- ing organisms in the distribution reservoirs. The growth of Asterionella in Ridgewood and Mt. Prospect reservoirs and its effect upon the quality of the water have been so fully de- scribed (report of Dr. Albert R. Leeds to the Department of City Works, Division of Water Supply, Brooklyn, 1897) that it is not necessary to again relate the details of its occurrence. That the growths of Asterionella continue to occur periodi- cally is shown by the diagram.
Asterionella is not the only odor-producing organism that develops in the distribution reservoirs. Anabaena, Synedra, Cyclotella and other forms are sometimes present in great abundance. The character of the water collected from the watershed of the Brooklyn supply is such as to furnish abun- dant nourishment for microscopic plant life, and organisms that in many water supplies appear in small numbers without having any noticeable effect on the character of the water develop in Brooklyn to an enormous extent.
This is emphatically true in the case of Synedra pulchella, a diatom that until recently has not been classed as an odor- producing organism. Like Asterionella, this diatom contains oil-globules, but the oily substance has not the same strong odor as the oil of Asterionella. Nevertheless, Synedra is capable of imparting an odor to water if present in sufficient numbers. The odor is not a characteristic one like that of Asterionella, Uroglena, Dinobryon, etc., and can be described by no more exact term than “vegetable.” The taste imparted to water by Synedra is perhaps more noticeable than the odor, | being somewhat “earthy,” as well as “vegetable.”
In few water supplies in this country is Synedra pulchella ever present in numbers greater than 5,000 per cu. cm. and,
36 GEORGE C. WHIPPLE
although a smaller number than this will make a water turbid, it requires about this number to produce a notice- able odor. In Brooklyn, however, the growths of Synedra have been much heavier, as may be seen from the diagram. On several occasions the numbers have reached 15,000 per cu. cm., and once as many as 20,000 per cu. cm. were observed. The water at such times has been very turbid, and has had the vegetable and earthy taste and odor just referred to.
The seasonable distribution of Synedra in the Brooklyn res- ervoirs is worth noting. In Mt. Prospect reservoir it has appeared regularly in the spring and fall, according to the usual mode of occurrence of the diatoms, but it has always appeared after the Asterionella growths in the spring and before the Asterionella growths in the fall. In Ridge wood its occurrence has been more variable. In 1899 there were heavy growths in basins 1 and 3 during the month of August.
Cyclotella is another diatom that, because of its limited occurrence, has been seldom known to cause trouble in water supplies. Yet in Ridgewood reservoir it is sometimes present in large numbers. Its growth has been usually of short dura- tion, but when present in numbers equal to 5,000 standard units per cu. cm. its aromatic odor could be distinctly recog- nized.
Two species of Melosira occur in the Brooklyn supply. Melosira granulata, the common free-floating form, is seldom present in sufficient numbers to cause trouble, though 2,000 or 3,000 per cu. em. are sometimes found. Melosira varians grows luxuriantly on the shores of Ridgewood reservoir, and constant scraping is required during the summer to keep the banks clean. During severe storms the filaments of Melosira become detached from the shores and are scattered through the water, and on one occasion the amount of vegetable mat- ter so detached was sufficient to impart a distinct taste to the water. Like Synedra pulchella, Melosira produces simply a vegetable, earthy and somewhat oily taste and odor, very different from the aromatic-fishy odor of Asterionella and Cyclotella.
Next to Asterionella, Anabaena has probably caused more trouble in the Brooklyn water supply than any other organ- ism. During the past two years it has appeared but once, but there are good reasons to believe that in the summer of
WORK OF MT. PROSPECT LABORATORY 37
1896, prior to the investigations of Dr. Leeds, the disagreeable odor of the tap water was due not so much to Asterionella as to Anabaena.
In July, 1898, Anabaena appeared in all the Ridgewood basins. In Basin 3 it did not develop to any extent. In Basin 1 it attained a maximum growth of 1.720 standard units per cu. cm. on August 19, and gave to the water its characteristic odor of moldy grass. In Basin 2, however, it developed to an enormous extent. On August 3 there were 24,000 standard units per cu.cm. From the last of July until early in Septem- ber the water in the basin was intensely turbid and had a green color. On quiet days a scum collected on the surface and drifted about with the wind. The water was entirely unfit for use, and the gates of the reservoir were kept closed. As soon as the organisms disappeared in the fall and the water had again assumed its normal condition, Basin 2 was emptied and cleaned, with the hope of preventing recurrence of such growths in the future. An examination of the deposit at the bottom of the reservoir showed that it was well seeded with the spores of Anabaena. Since that time there has been no further development of this organism.
In September, 1898, a phenomenally large growth of Scenedesmus occurred in Mt. Prospect reservoir, the water at one time containing 25,800 standard units per cu. cm. This organism, in the numbers ordinarily found, causes no odor, but on this occasion the water had a distinct vegetable and aromatic odor and taste. The growth continued for several weeks.
There are several other organisms that deserve mention, because they occur in larger numbers in the Brooklyn water than in most water supplies. Dictyosphaerium, Eudorina, Pandorina and Volvox are often present in numbers of 500 standard units per cu. em. Clathrocystis is not often found in Ridgewood reservoir, but in Mt. Prospect reservoir it has been as high as 1,440 standard units per cu. cm. As a rule the Brooklyn water contains comparatively few protozoa, but Mallomonas has been observed as high as 660 per cu. cm. in Ridgewood reservoir, and Cryptomonas has been as high as 2,000 per cu. em. in Mt. Prospect reservoir. Chlamydomonas has been found occasionally.
To the water consumers of Brooklyn, however, the im- portant fact is not the number of organisms in the distribution
38 GEORGE C. WHIPPLE
reservoirs, but the number present in the tap water in the city. Prior to the construction of the by-pass at Ridgewood the organisms that developed in the reservoir found their way as a matter of course to the service taps of the consumers. But by using the by-pass it has been found possible to so regulate the distribution of the water that very few organisms reach the consumer. Guided by the frequent and regular mic- roscopical examinations made at Mt. Prospect laboratory, the engineer has directed one or more basins to be isolated whenever it was found that odor-producing organisms were developing in them, the water meanwhile being delivered through the by-pass direct from the force mains to the dis- tribution pipes. It has been found possible also to isolate Mt. Prospect reservoir and pump directly into the pipes when growths of organisms made it seem advisable. The beneficial effect of this management can be illustrated by the following comparison.
At the time when Dr. Leeds made his report on the con- dition of the water, i. e., from November, 1896, to February, 1897, Asterionella was present in the distribution system as follows:
No. per cu. em.
idvewoud Teservoir; asin 1 sss ve ee Ses Bevake Wea ela 3to 48 Se a SE een hs ny chi at) Taare ce erate ne eae Peden 21)
3 cs CLT) Ree aA BALD Seta bg re ae Shs | 2,608 ** 4,648
ME, SPCOSPOGh TESOLVOIE! Aeuee ety fonts. clse ciemek Sen ees Gace 4,808 ** 8,640 Tap supplied from Ridgewood, Basing Wiad, Rite! eves a erent | [SS cy Ags SRE ee MnP RAE . 1,240 * 8,800
0 A OMG; (Prospect Teservoir .(.! 2 6/522 le win bea 2,400 ‘* 7,460
During November and December, 1899, the corresponding figures were as follows: No. per cu. em.
Ridsewooud Teseryoir, Basin d 636 8s 2ik Ae ales oe nen oc 5,600 to 27,280
Fe a RON AOE Tie We Ls tA Nae ay, Ohne Orns 8
ce sé AiG Ns Stirs, GL eNe ero EAR ICON OrIRe Opes 16 Mii WProspaet TESCLVOMN) cen seis ccs eeisemclde sc. e's + sels sake 6,512 ‘* 24,960 Tap ordinarily supplied from Ridgewood, Basin 1 and 2.. 0 Taps supplied from Ridgewood, Basin 3.................. Os 16 Tap ordinarly supplied from Mt. Prospect reservoir ...... Ss 56
During November, 1896, and February, 1897, the water supplied to the city from Basin 3 and from Mt. Prospect reservoir had a very disagreeable taste and odor, due to the presence of Asterionella, but during November-December, 1899, the water in the city had no odor due to Asterionella, even though that organism was far more abundant in Ridge-
WORK OF MT. PROSPECT LABORATORY 39
wood and Mt. Prospect reservoirs than it had been during the winter of 1896-97. This freedom of the tap water from Asterionella was due to the use of the by-pass, the sections of the city that are ordinarily supplied from Ridgewood Basins 1 and 2 and from Mt. Prospect reservoir being supplied with water direct from the Ridgewood force mains.
40) WORK OF MT. PROSPECT LABORATORY
EXPLANATION OF PLATES
Plate I
Exterior view of Mount Prospect Laboratory, Brooklyn Water Works.
Plate II Plan of main floor, Mount Prospect Laboratory.
Plate IIT View of a portion of the Chemical Laboratory.
Plate IV
Variations in numbers of microscopic organisms in the Brooklyn Reservoirs, November, 1897, to February, 1900.
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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
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PLATE XI
PLATE XII
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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-