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Jagger. ye ete gees Pe caeaed z anathinn nan). ed A thet d qvse¢ sniper LLY Pere nd bre, hal aan ‘ gq “7 ede bd 4@¢4 Jj 949 ee heme 778 a= ae, | tT rience Pde vy é Bh Suh as” Sar je 3 Fete ath Hat * Y i Sey / i my arse bs 3, MY (leg aMRcee NM Mare TAS nf hN : ¥ Rata Migs Mae ah Stl 5, r ay i ‘ A a, See a } ¢ , ' » & \ 1 < % DEPARTMENT OF MARINE BIOLOGY aan OF ; ye CARNEGIE INSTITUTION OF WASHINGTON ©) ALFRED G. MAYER, DIRECTOR & ~ PAPERS FROM THE TORTUGAS LABORATORY 3 OF THE CARNEGIE INSTITUTION OF WASHINGTON VOLUME V a MAR 1° 1923 4 Peete @S4] Betas! wars A WASHINGTON, D. C. PUBLISHED BY THE CARNEGIE INSTITUTION OF WASHINGTON IQI4 ha at Oy rai oe ee 4 if ” V4 u i= eed te, ge ee Y / Le , See Cog oral a ga Der Nh =e rat Co &- eet ee le Fy A /loce B po - (] ND ' DEPARTMENT OF MARINE BIOLOGY Snir ot 2 0.4 f : OF CARNEGIE INSTITUTION OF WASHINGTON ALFRED G. MAYER, DIRECTOR PAPERS FROM THE TORTUGAS LABORATORY OF THE CARNEGIE INSTITUTION OF WASHINGTON VOLUME V WASHINGTON, D. C. eee PUBLISHED BY THE CARNEGIE INSTITUTION OF WASHINGTON IQI4 CARNEGIE INSTITUTION OF WASHINGTON PUBLICATION NO. 182 Copies of this Book 4 ; were first issued | a APR1 1914 PRESS OF THE NEW ERA PRINTING COMPANY LANCASTER, PA. CONTENTS. I. In Memoriam: George Harold Drew. By ALFRED G. MAYER....1 plate II. On the Precipitation of Calcium Carbonate in the Sea by Marine Bacteria, and on the Action of Denitrifying Bacteria in Tropical and Tem- perate Seas. By Gi H. DREW: n 2 2.06 a swe 2 maps, 4 figures III. Preliminary Remarks on the Geology of the Bahamas, with Special Refer- ence to the Origin of the Bahaman and Floridian Oolites. By T. WAM AWD! VAUGHAN) S c.505 catches ctelere ciate e ofasoim singe eee emia IV. Building of the Marquesas and Tortugas Atolls and a Sketch of the Geologic History of the Florida Reef Tract. By T. WAYLAND VAUGHAN.. V. Some Chemical Characteristics of Sea-water at Tortugas, Florida, By Mp eet LOL Bitar no tut tornpatete ata, o atacate aoe oy agetm a wens: wieatis a0 sere ay antns cata VI. Observations upon the Growth-rate and Ecology of Gorgonians. ByL.R. CARNE sh aiel erarev vere eere eee ies ote ic ohalonel ote or ane omy srarapatayere 2 plates VII. Growth-Changes in Brittle-Stars. By H. L. CLARK........... 3, plates VIII. The Early Influence of the Spermatozoan upon the Characters of Echinoid harve, “Bye Ty eNNENE cas... «ce eitearsea© os'ste ne, II figures IX. Studies of Jamaica Echini. By RoBERT T. JACKSON......... 21 figures X. The Spermatogenesis of the Mongoose; and a Further Comparative Study of Mammalian Spermatogenesis, with Special Reference to Sex Chromosomes. By H. E. JORDAN............. 1 plate, 9 figures XI. Bryozoa of the Tortugas Islands, Florida. By R. C. OsBuRN....23 figures Page. 1-6 7-45 47-54 55-67 69-78 79-90 QI-126 127-138 139-162 163-180 181-222 iil E IN MEMORIAM. GEORGE HAROLD DREW, 1881-1913. One plate. GEORGE HAROLD DREW. (1881-1913.) George Harold Drew, only son of George Drew, of Devonshire, England, was born on October 23, 1881, and died suddenly on January 29, 1913. From early boyhood chemistry and microscopy fascinated him, and while yet a child he studied in a disused stable on his father’s place, which was made to serve as a rough laboratory. Until the age of ten he was taught by his mother, and then he was sent to New College, Eastbourne, Sussex. Being deeply interested in the scien- tific side of medicine, he entered Cambridge University, where he became a scholar of Christ’s College, and gained the distinctions of Exhibitioner, Foundation Scholar, and Prizeman, afterwards becoming Schreiner Uni- versity Scholar, and a student at Exeter Hospital. During his five years at Cambridge he took the entire medical course, but afterwards decided to devote his life to research, inclination leading him to choose the scientific side of his profession rather than to engage in its active practice. After leaving Cambridge, he served for a year as University Scholar of St. Mary’s Hospital, London, and subsequently, after working for a short time at the Port Erin Marine Laboratory, he went to Plymouth and associ- ated himself with the Marine Biological Laboratory, wherein most of his scientific work was accomplished. He was one of those chosen by Captain Scott to serve upon the scientific staff of the British Antarctic Expedition, but he declined the honor, preferring not to break the continuity of his studies at Plymouth. From 1910 until 1912 he held a Beit Memorial Fellowship for Medical Research in cancer; in the summer of 1912 he was appointed John Lucas Walker research student in the University of Cambridge, and on January I, 1913, he became a Research Associate of the Department of Marine Biology of the Carnegie Institution of Washington. His connection with the Carnegie Institution of Washington began in 1911, when he accompanied the expedition to Jamaica, afterwards studying at the Tortugas, and upon a second visit to America in 1912 he spent a month at the temporary laboratory established at Golding Cay, Andros Island, Bahamas. He was to have been a member of the projected expedition to Torres Straits, Australia, in 1913, and his preparations were being matured at the time of his sudden death. The saddest losses that the world sustains are often those it heeds the least, and all who knew him realize that this young man, with his keen 3 4 Papers from the Marine Biological Laboratory at Tortugas. insight for research, his broad culture, and well-rounded special training, was destined to become one of England’s great men of science—a position which his rare personal charm would not only have graced, but would have rendered him powerful for advancing the intellectual welfare of his country. In 1910 he carried out some interesting observations upon the repro- duction and early development of the kelp (Laminaria) of the coast of Devonshire, proving that there is an alternation of generations in the life cycle of this seaweed; for the free-swimming sexual cells fuse in pairs and then settle down upon the bottom and develop into a chain of cells, any single cells of which may develop into a Laminaria plant. His published works cover a period of only four years, yet in this brief time he was the author or joint author of fifteen papers. His earliest paper, in 1909, is upon parasitic and other diseases of fish, in which he describes various forms of tumors and cancers. This paper was followed by one of an experimental nature written in association with de Morgan as its joint author, and it was shown that, in Pecten, injurious bodies implanted in the tissues soon become surrounded by an agglutinated layer of blood-corpuscles and these are replaced by a mass of fibrous tissue which encapsules the foreign body very much as such bodies are incased by mesenchyme in injured vertebrates. In 1911 Drew announced that, as a result of injury, cysts lined by columnar ciliated epithelium could be formed from the fibroblasts in Pecten. He also found that the blood corpuscles of Lamellibranchs are capable of ingesting and destroying bacteria, and that if the animal be wounded so that blood escapes, some of the corpuscles adhere to the cut surfaces and then send out slender processes which fuse with those from other corpuscles, thus forming a net-work the fibers of which finally contract and close the wound. He also found that the repeated application of a strong solution of iodine to a circumscribed area of the skin of Fundulus produces an inflammatory tumor having some relation to certain abnormal growths seen in nature in fishes. While at Plymouth the subject of the cause and nature of tumors and cancers engaged his major interest, and he did much histological work upon the fate or growth of transplanted tissues. His last work was upon the culture of living tissues from the frog and the dogfish in plasma outside the body of the animal. In this study he achieved decided success, having with his usual ingenuity devised an improved technique which enabled him to grow tissues upon microscope slides free from bacterial contamination. The paper describing these results was completed only a few days before he died and is published in the Journal of Pathology and Bacteriology, Cambridge. Some of his most interesting work, however, was done while associated with the Department of Marine Biology of the Carnegie Institution of Washington. George Harold Drew. 5 In 1910 Sanford, and also Vaughan, published the conclusion that a considerable portion of the calcareous muds in the bays and sounds of southern Florida were precipitated out of the sea-water in some unknown manner. It remained for Drew, in 1911, to discover that there is in the warm surface waters of the West Indian and Florida region, and especially in the limestone mud itself, a bacillus which deprives the sea-water of its nitrogen, thus causing the calcium to combine with the dissolved carbon dioxide and to form the finely-divided limestone mud so characteristic of coral-reef regions. Drew isolated this bacillus and found that it became inactive in even moderately cold water, and thus it functions only in warm or tropical seas, thriving best at depths of less than 100 fathoms. In the surface waters of the Bahamas and Florida it is the most abundant marine bacillus. Drew hoped to continue these studies and to extend them to the Pacific, for this calcium precipitation is an important factor, and has resulted in the formation of vast beds of limestone apparently far greater in bulk than that formed by corals. The complex problem of the chemical balance of the constituents of sea- water and their solvent powers under various conditions was being enthusi- astically considered by Drew, and had he lived it was the hope of this Labo- ratory that he might have had exceptional opportunities to continue these studies—for truly he was the exceptionally brilliant man, for the coming of whose like again the world must wait, we fear, for many years, for there are but few in each generation who are gifted with his rare genius for research. ALFRED GOLDSBOROUGH MAYER. BIBLIOGRAPHY. 1. Some notes on parasites and other diseases of fish, in Parasitology, vol. 2, No. 3, p. 193, 1909; also vol. 3, No. I, p. 54, 1910. Also a review of the same in Journal Marine Biol. Association, Plymouth, vol. 9, No. 2, p. 246, I9II. 2. The reproduction and early development of Laminaria digitata and Laminaria sac- charina, in Annals of Botany, vol. 24, p.177, 1910. Also reviewed in Journal Marine Biol. Association, Plymouth, vol. 9, No. 2, p. 245, I9II. 3. Some points in the physiology of Lamellibranch blood-corpuscles, in Quart. Journal Microscop. Sci., vol. 54, pp. 605-622, 1910. 4. A table showing certain culture characteristics of some of the commonest bacteria found in the laboratory tanks at Plymouth, in Jour. Marine Biol. Association, vol. 9, No. 2, pp. 161-163, I9II. 5. The action of some denitrifying bacteria in tropical and temperate seas, and the bacterial precipitation of calcium carbonate in the sea, in Journal Marine Biol. Association, vol. 9, No. 2, pp. 142-155, 1911. Also in the Report of the Department of Marine Biology, Year Book of the Carnegie Institution of Washington, No. 10, pp. 136-141, IQII. 6. Some cases of new growths in fish, in Journal Marine Biol. Association, Plymouth, vol. 9, No. 3, pp. 281-287, I plate, 1912. 7. A note on some attempts to cause the formation of cytolysins and precipitins in certain invertebrates, in Jour. Hygiene, vol. 11, No. 2, 1911. 8. Experimental metaplasia. 1. The formation of columnar ciliated epithelium from fibroblasts in Pecten, in Journal of Experimental Zoology, vol. 10, pp. 349-374, 3 plates, I9II. 14. 15. Papers from the Marine Biological Laboratory at Tortugas. . A note on the application of Giemsa’s Romanowsky stain to the blood and tissues of marine invertebrates, in Parasitology, vol. 4, pp. 19-21, I9II. . An experimental investigation of the cytological changes produced in epithelial cells by long-continued irritation, in Journal of Pathology, vol. 17, 1912. . Report upon the formation of calcium carbonate through bacterial action in the tropical Atlantic, Report of the Department of Marine Biology, Year Book of the Carnegie Institution of Washington, No. 11, pp. 136-144, 1913. . On the culture in vitro of some tissues of the adult frog, in Journal of Pathology and Bacteriology, vol. 17, pp. 581-583, plates 44-46, 1913. . The origin and formation of fibrous tissues as a reaction to injury in Pecten maximus, by G. H. Drew and W. de Morgan, in Quart. Journal Microscop. Sci., vol. 55, pp. 595- 610, plate 24, 1910. Also a review of the same in Journal of the Marine Biol. Association, vol. 9, No. 2, p. 595, I9II. Note on the abnormal pigmentation of a whiting infested by trematode larve, by F, W. Gamble and G. H. Drew, in Journal Marine Biol. Assoc., vol. 9, No. 2, p. 243. IgII. On the pricipitation of calcium carbonate in the sea by marine bacteria, and on the action of denitrifying bacteria in tropical and temperate seas, in Papers from the Marine Biological Laboratory at Tortugas, Publication No. 182, Carnegie Institution of Washington, 1913. II. ON THE PRECIPITATION OF CALCIUM CARBONATE IN THE SEA BY MARINE BACTERIA, AND ON THE ACTION OF DENITRIFYING BACTERIA IN TROPICAL AND TEMPERATE SEAS. By G. HAROLD DREW. Two maps and four figures. TABLE OF CONTENTS. Page Introduction 54 302<5 Acts ASSES Bae a ee eee eee 9 General, considerations and: previous: Work... «.6/< ); about 5.0 grams; sea-water, 1,000.0 c.c. Calcium malate is only slightly soluble in water, so can be added in excess. Gran used distilled water and added 30 gm. sodium chloride per liter, but in these experiments sea-water has been used instead. II. Calcium succinate medium: Calcium succinate (CHK E00 >C ): 2,0 grams; potassium nitrate (KNOs), 0.5 gram; sodium phosphate (NazHPOs, 12H:0), 0.25 gram; sea-water, 1,000.0 C.c. 20 Papers from the Marine Biological Laboratory at Tortugas. This medium was boiled and filtered before sterilization to remove the slight precipitate of calcium phosphate. It was found that this medium with the addition of the phosphate gave a more vigorous growth than if it was omitted. III. Calcium acetate medium: Calcium acetate (Ca(CH3sCOO),), 5.0 grams; sodium phosphate (NazHPO,., 12H2O), 0.25 gram; potassium nitrate (KNOs), 0.5 gram; sea-water, 1,000.0 c.c. Boiled and filtered before sterilization to remove precipitate of phos- phate. IV. Peptone calcium acetate medium: Calcium acetate (Ca(CH;COO):), 5.0 grams; peptone (Witte’s), 0.2 gram; potassium nitrate (KNO;), 0.5 gram; sea- water, 1,000.0 c.c. Media II, III, and IV were also made up with the addition of 0.2 gm. of magnesium tartrate per I,000 c.c. The fluid media were made up in 1,500 c.c. resistance glass flasks, and 1,000 c.c. of medium were used for each culture. For other purposes a simple solution of peptone in sea-water was em- ployed (2 gm. to 1,000 c.c.), and media were also used consisting of this peptone solution with the addition of 0.5 per cent of various carbohydrates, such as cane sugar, dextrose, levulose, mannite, lactose, etc., with sufficient neutral red solution to color them, in order to test the acid-forming proper- ties of the bacteria in the presence of carbohydrates. The ordinary “‘Koch”’ steam sterilizer and iron oven for dry-heat sterilization were used, and gasoline cooking stoves were found to be the most satisfactory source of heat. It was found an advantage to use Petri dishes with porous earthenware covers which enabled the water of con- densation to evaporate partially; the evaporation could be checked at any time by covering the dishes with a bell jar lined with wet filter paper. It was usually found necessary to keep all cultures on tables with their feet standing in dishes of kerosene, in order to prevent the attacks of ants and other insects. In all other respects ordinary bacteriological routine was followed, and the methods need not be further particularized here. The reduction of the nitrate to a nitrite in fluid culture media was tested for by the addition of 5 c.c. of 10 per cent sulphuric acid and 2 c.c. of a 1 per cent solution of metaphenylene diamine hydrochloride to 25 c.c. of the culture. The production of a brown coloration (due to the formation of Bismarck brown) is an indication of the presence of a nitrite, and is an extremely delicate reaction. The diphenylamine and brucine sulphate reactions were also used when testing for the presence of nitrates. The formation of ammonia was tested for by the addition of 5 c.c. of IO per cent potassium hydrate and 5 c.c. of Nessler’s reagent; the white precipitate formed on the addition of the potassium hydrate does not appreciably interfere with the test, though it renders it somewhat less delicate. On the Precipitation of Calcium Carbonate. 21 Under expeditionary conditions, and in the absence of the somewhat elaborate apparatus that would be necessary in order to estimate chemically the amount of denitrification in cultures, it was only possible to compare the rate of denitrification in different cultures by noting the time taken for the first appearance of the nitrite reaction and the time taken for all trace of nitrite or nitrate to disappear. It seems that the rate of denitrification in culture media inoculated with equal volumes of samples of sea-water must be a function of the number of bacteria in the sample, the temperature at which the cultures are grown, and the specific power of denitrification of the individual species of bacteria. Considering the rapid multiplication of bacteria when the food supply is plentiful, up to a maximum determined chiefly by the accumulation of the waste products of their own metabolism, it appears that the factor of the number of bacteria in the sample may be neglected within the limits of these experiments. For example, the number of bacteria in 1,000 c.c. of Gran’s medium at the end of 24 hours would probably be much the same whether it were inoculated from a sample con- taining 8 or 16 bacteria per I c.c.; similarly it was a matter of experience that the first trace of nitrite formation was observable at about the same time, whether 5 or 10 c.c. of a given sample had been used for inoculation. Consequently it would appear that for purposes of comparison, and within the limits of the experiments described, if the temperature be the same for the cultures compared, the rate of denitrification is a measure of the specific denitrifying power of the particular species of bacteria. In the work on the bacterial precipitation of calcium carbonate, the precipitate (which was often so fine as to tend to remain in suspension) was usually obtained by centrifuging. It was either preserved in small bottles with some of the culture fluid, or else washed first with distilled water and then with absolute alcohol, and finally allowed to dry. These precipi- tates were sent to Dr. F. E. Wright, of the Geophysical Laboratory of the Carnegie Institution of Washington, who with great kindness reported on their mineralogical properties. INVESTIGATION OF SAMPLES OF SEA-WATER TAKEN OFF PORT ROYAL, JAMAICA. The work at Port Royal was done in May tg11, but was of a very pre- liminary nature. It was necessary to depend on a sailing-boat for obtaining the samples; yet, owing to the remarkable regularity with which an on-shore wind springs up every morning, but little difficulty was experienced from this cause. No apparatus for obtaining deep samples was available, but samples were taken from a depth of 6 fathoms by means of a bottle from which the stopper was pulled by a line and then allowed to fall back into place. A measurement of the rate of denitrification in fluid culture media inoculated with samples of sea-water was made, but isolation of the bacteria on solid media was not attempted. The following method was employed: 22 Papers from the Marine Biological Laboratory at Tortugas. Samples of sea-water were collected in sterilized stoppered bottles from the surface and from depths of 3 and 6 fathoms from positions about 5 miles from shore, where, from a consideration of the wind and tide, the water was probably under truly oceanic conditions and unaffected by the neighboring land. 10 c.c. of these samples were added to 1,000 c.c. of Gran’s medium. The cultures were kept in a moderate light and the temperature varied from 25° to 31.5°C. The average temperature during the growth of each culture was noted. In a typical culture made from surface water, and for which the average temperature was 29° C., the first indication of the formation of a nitrite, as given by the metaphenylene diamine reaction, appeared after 27 hours; after 38 hours the brown color produced in this reaction was very intense, the culture became cloudy, and on testing with Nessler’s reagent slight ammonia formation was apparent. After 48 hours the culture became very cloudy and a scum of bacterial growth developed; the nitrite and ammonia reactions remained unaltered. After 63 hours the nitrite reaction was somewhat less marked, the ammonia reaction was unaltered, and bubbles of gas began to appear. After 72 hours many bubbles of gas were being produced, and the nitrite and ammonia reactions were very slight. After 86 hours the bubbling had ceased, and no nitrite or ammonia was present in the culture. Testing the culture for nitrates by the brucine and diphenylamine reactions then showed that no nitrate was left in the solution. In the absence of a gas-analysis apparatus the nature of the gas evolved could not be determined, but considering that it was non-inflammable, did not turn lime-water milky, and that the nitrate originally present had been destroyed, it seems strongly probable that this gas was pure nitrogen. Thus in 86 hours 0.5 gm. of potassium nitrate had been decomposed by bacterial growth. Ifa further 0.5 gm. of potassium nitrate were then added, it was rapidly decomposed, and this could be repeated many times until the other constituents of the culture medium were used up. It was found that the rate of denitrification varied somewhat with the temperature, and that in cultures kept at a temperature of between 10° and 12° C. no growth or denitrification occurred. The denitrification was always more rapid in cultures from water taken from a depth of 3 or 6 fathoms than from the surface. It was also more rapid with samples taken from the thick muddy water of a mangrove swamp, where organic matter was plentiful. The bacteria present in the cultures were very minute, actively motile bacilli with rounded ends. An abstract of the behavior of a few of the cultures is given below. I. Sample collected 5 miles south of Port Royal, wind southeast, force 4, tide rising. Sample taken from surface. 1,000 c.c. of Gran’s medium was inoculated with 10 c.c. of sample. After 20 hours a slight cloud developed in the culture and faint nitrite reactions were given. On the Precipitation of Calcium Carbonate. 23 After 36 hours a dense cloud developed in the culture and strong nitrite reactions were given. After 60 hours a dense cloud and scum developed in the culture and strong nitrite and faint ammonia reactions were given. After 70 hoursa dense cloud, scum, and bubbles developed in the culture and faint nitrite and faint ammonia reactions were given. After 84 hours culture was less cloudy, with much scum, no nitrite or nitrate reaction, very faint ammonia. Average temperature at which the culture was grown, 30° C. 2. Sample collected from same spot under similar conditions, from a depth of 3 fathoms. 1,000 c.c. Gran’s medium was inoculated with 10 c.c. of sample. After 20 hoursa slight cloud developed in the culture and faint nitrite reaction was given. After 27 hours a denser cloud developed in the culture and strong nitrite and faint ammonia reactions were given. After 38 hours a dense cloud and scum developed in the culture and strong nitrite and faint ammonia reactions were given. After 48 hours a dense cloud and scum developed in the culture and moderate nitrite and faint ammonia reactions were given. After 63 hours a moderate cloud, thick scum, and bubbles developed in the culture and faint nitrite and faint ammonia reactions were given. After 72 hours a slight cloud and thick scum, no nitrite or nitrate, and very faint ammonia reaction. Average temperature at which the culture was grown, 29° C. 3. Sample collected from a spot 6 miles south of Port Royal, wind ESE., force 4, high tide (slack). Taken from surface. 1,000 c.c. Gran’s medium was inoculated with 10 c.c. of the sample. After 20 hours a slight cloud developed in the culture, no nitrite reaction was given. After 27 hours a slight cloud developed in the culture, faint nitrite reaction was given. After 38 hours a dense cloud developed in the culture, strong nitrite and faint ammonia reactions were given. After 48 hours a dense cloud and scum developed in the culture, strong nitrite and faint ammonia reactions were given. After 63 hours a dense cloud and scum developed in the culture, moderate nitrite and faint ammonia reactions were given. After 72 hours a moderate cloud, scum, and bubbles developed in the culture, very slight nitrite and faint ammonia reactions were given. After 86 hours a moderate cloud and scum, no nitrite or nitrate and very faint ammonia reaction. Average temperature at which the culture was grown, 29° C. 4. Sample taken from surface water of the large mangrove swamp lying northwest of Port Henderson. 1,000 c.c. of Gran’s medium inoculated with 10 c.c. of sample. After 20 hours no cloud or nitrite reaction. After 24 hours slight cloud and slight nitrite reaction. After 40 hours strong cloud and scum, strong nitrite and slight ammonia reaction. After 75 hours cloud, scum, and bubbles, no nitrite or nitrate and slight ammonia reaction. Average temperature at which the culture was kept, 30° C. 5. Subculture from culture (1). 1,000 c.c. Gran’s medium inoculated with § c.c. of culture (1), and kept at a temperature of 10° C. to 12° C. by means of ice. After 100 hours the culture was quite clear and gave no nitrite reaction. It was then removed from the ice and kept at the room temperature, which averaged 30° C. After 107 hours a dense cloud developed in the culture and strong nitrite and faint ammonia reactions were given. After 120 hours a dense cloud, scum, and bubbles developed in the culture and moderate nitrite and faint ammonia reactions were given. 24 Papers from the Marine Biological Laboratory at Tortugas. After 131 hours a faint cloud, scum, and bubbles developed in the culture; very faint nitrite and faint ammonia reactions were given. After 146 hours a faint cloud and scum, no nitrite or nitrate, and very slight ammonia reactions were given. Twenty cultures were made from samples of water taken well out to sea from Port Royal, and the process of denitrification followed through with each. All gave very similar and consistent results, but the rate of denitri- fication decreased rapidly with the temperature at which the cultures were grown; thus at an average temperature of 27° C. the first trace of the nitrite reaction appeared after about 40 hours, and denitrification was complete after about 100 hours. The results of precisely similar experiments that I made with samples of water taken from the English Channel near Plymouth in the autumn of 1909 showed that there the process of denitrification was very much slower, and was never complete at the room temperature (17° C.). The first trace of the formation of a nitrite in cultures in the modified Gran’s medium, as detected by the metaphenylene diamine reaction, occurred about the fifth day, and a large proportion of the nitrite and nitrate always remained, even in the oldest cultures. In similar cultures incubated at 30° C., denitrification was complete by the eighth day at earliest, but uniformly consistent results were not obtained, as in some of the cultures complete denitrification was never obtained, even after several months. It would thus appear that even under similar temperature conditions the marine bacteria in the seas off Jamaica are much more active in causing denitrification than those found in the English Channel, and since the rate of denitrification is a function of the temperature, it follows all the more that the destruction of nitrates by bacterial agency in the seas around Jamaica must be far in excess of that occurring in the cooler waters of the English Channel. THE INVESTIGATION OF SAMPLES OF SEA-WATER TAKEN AROUND THE DRY TORTUGAS. The Dry Tortugas consist of a group of eight small keys, the largest of which (Loggerhead Key) is only about three-quarters of a mile long by one- eighth of a mile wide. They are situated about 150 miles from the main- land of Florida, and form the extreme western end of the chain of the Florida Keys. The 100-fathom line lies some 30 miles to the south and southwest of the islands, and then trends round in a northwest direction; beyond the 100-fathom line the depth increases with moderate rapidity until depths of from 1,000 to 1,400 fathoms are reached. To the east, northeast, and north as far as the coast of Florida, the water is shallow, the soundings showing from 20 to 30 fathoms in most places. Beyond the 100-fathom line to the southward the influence of the Gulf Stream begins to make itself felt, though the region of maximum current velocity here lies nearer the coast of Cuba. The Tortugas Keys are of purely coral forma- tion, consisting entirely of broken shell and coral sand, with no soil. The On the Precipitation of Calcium Carbonate. 25 greatest elevations are the hurricane ridges, which are not more than 15 feet above sea-level, and during a hurricane the islands are sometimes completely submerged. There is no vegetation on the smaller keys, but Loggerhead Key, on which the Marine Biological Laboratory of the Car- negie Institution of Washington is situated, is partially covered with a growth of bushes and coarse grass. There is no fresh-water supply on the islands. From these considerations it is obvious that the risk of contamination of samples of sea-water taken a few miles from the Keys through land bacteria is very small, and that such samples may be taken as being truly oceanic, The motor yacht Anton Dohrn and smaller motor boats made the col- lection of samples an easy matter, and the well-equipped laboratory made possible fuller investigations than those attempted in Jamaica. A number of cultures were made in Gran’s medium under conditions exactly comparable to those made at Port Royal, and the rate at which the process of denitrification proceeded was observed. The results agreed almost exactly with those obtained at Port Royal, so need not be described in detail. It thus seems that the denitrifying power of the bacteria in the seas around the Tortugas is the same as that of those around Jamaica. Cultures were also made on various solid media, and pure cultures of the various species of bacteria were isolated by plating in Petri dishes with peptone agar. Samples of surface water taken from various positions around Tortugas as far as possible removed from influence of the land, and collected on sunny days, gave an average count of 14 colonies per I c.c. of sample. Counts of several plates from the same locality, and from different localities, showed a somewhat remarkable agreement as to the number of colonies present, the highest count ever obtained being 20 and the lowest 8 per 1 c.c. Allowing for experimental error, this shows great uniformity in the distribution of bacteria in the sea around Tortugas. The colonies appeared to be of two kinds when grown on peptone agar, one much more plentiful than the other. Subcultures made from these colonies in Gran’s medium showed that the bacteria forming the most com- mon type of colony produced an active denitrification, while the others grew very slowly in this medium and produced no denitrification. The characteristics of the denitrifying form are as follows: On the potassium malate, or peptone agar media, colonies are visible as minute white specks after 6 to 8 hours when the room temperature averages 29.5° C. After about 18 hours the colonies are well developed; they are white in color, circular but with finely irregular outline, and have a granular appearance. Superficial colonies are much elevated at first, but as growth proceeds they spread rapidly over the surface of the agar. Deep colonies remain small, circular, and discrete. Growth is somewhat more rapid on peptone agar than on the potassium malate agar, and the older colonies develop a brownish tinge in the center when growing on the former medium. On gelatin peptone (5 per cent 26 Papers from the Marine Biological Laboratory at Tortugas. peptone in sea-water and kept at between 20° and 25° C. to insure the medium remaining solid), growth was very slow; in stab cultures growth proceeded slowly from the surface downwards, leaving a funnel-shaped depression of liquefied gelatin. Acid formation occurs in dextrose, levulose, mannite, and cane sugar, but not in lactose media. Growth is inhibited at a temperature of 10° C., but takes place slowly abn oe. Growth is much retarded by exposure to bright sunlight, but the bacteria are not killed by a Io hours’ exposure. The bacteria are facultative anaérobes, but growth under anaérobic conditions is very slow. In Gran’s medium growth is rapid, but no growth occurs if the potassium nitrate be omitted, or if the calcium malate be replaced by calcium carbon- ate. Growth in a pure solution of peptone in sea-water is very slight, but becomes abundant if potassium nitrate be added, when denitrification quickly ensues. The most rapid growth was produced in sea-water con- taining 2 per cent peptone, I per cent potassium malate, and 0.5 per cent potassium nitrate, and in this clear medium a slight floccular precipitate, presumably of calcium salts derived from the sea-water, was soon formed. Growth was also rapid at first in a solution of 5 per cent potassium malate and 0.5 per cent potassium nitrate in sea-water, but growth apparently ceased in this medium after a few days and denitrification was never com- plete; a slight precipitation occurred and the solution was found to have very definitely increased in alkalinity. This bacterium does not appear to have been previously described, and I propose for it the name of Bacterium calcis, owing to its power of pre- cipitating calcium carbonate from solutions of calcium salts. This point will be dealt with later in the paper. The characteristics of the scarcer non-denitrifying form of bacterium found on the agar plates are as follows: Growth on the potassium malate agar medium is very slow and indefi- nite. On peptone agar growth is somewhat slower than in the case of the denitrifying form. On the surface, circular cream-colored colonies are formed, having a brownish center; the edges are smooth and regular, and the colony remains discrete and does not tend to spread over the surface. The deep colonies are smaller and usually ovoid in shape, and of a somewhat darker color than those on the surface. No growth was obtained on gelatin media. Acid formation as shown by the neutral red reaction occurs in dextrose and lzvulose, but not in cane sugar, lactose, or mannite media. Growth takes place slowly at 10° C. No visible growth occurred at 0° C., but cultures were not killed by 24 hours’ exposure to this temperature. Growth is retarded by light, and cultures are killed by 4 hours’ exposure to bright sunlight. The bacterium is a strict aérobe. On the Precipitation of Calcium Carbonate. ag Free growth takes place in Gran’s medium, but develops much slower than in the case of the denitrifying form. No growth occurs if the potassium nitrate be omitted entirely, but takes place freely if a mere trace in excess of that normally present in the sea-water be added, though no denitrification results. Attempts were made to discover whether this bacterium had any nitrifying or denitrifying action in various culture media, but uniformly negative results were obtained. Nitrites were neither oxidized to nitrates nor reduced to ammonia or free nitrogen, and ammonia salts were un- affected. No growth was obtained in any culture medium that did not contain at least a trace of nitrates, so it was not practicable to ascertain whether the bacterium had a nitrifying action without the necessary facili- ties for quantitative work. On one occasion samples were obtained from various depths up to 90 fathoms at a point near the Gulf Stream region, 25 miles south of Tortugas. Exhausted glass flasks with capillary necks which could be broken off at the required depth were used for the purpose. These samples were plated in the peptone agar medium and counted, with the following average results: Depth in Denitrifying | Non-denitrifying | No. of colonies fathoms. orms (Bacterium Tae. developing from calcis). I c.c. of sample. o 9 2 II Io 25 4 29 40 2 2 4 oe 5 3 8 90 5 6 It These figures are probably not very reliable, especially for the greater depths, since it is possible that many of the bacteria were killed by the sud- den reduction of pressure to which they were exposed as the water entered the exhausted bulb. INVESTIGATION OF SAMPLES FROM A POINT 70 MILES WEST OF USHANT ISLAND, FRANCE. This spot was chosen as it is sufficiently far out in the Atlantic to be largely removed from the influence of the English Channel water. The object was to investigate truly oceanic bacteria, and previous work in 1909 had shown that the bacterial flora of the Channel water was relatively very complicated, probably owing to the presence of littoral forms. The Marine Biological Association of the United Kingdom very kindly sent their steamship Oithona from Plymouth for this work, and gave me every facility both on board and in their laboratory. As in Tortugas, the deep samples were collected in exhausted glass flasks, and accordingly, as previously ex- plained, the results obtained from the deep samples can not be considered to possess any very great degree of accuracy. Attempts were made to plate the sample in peptone agar on board the boat, but the result was not satisfactory, as owing to the motion of the boat 28 Papers from the Marine Biological Laboratory at Tortugas. the jelly set in irregular waves and lumps. Consequently the samples were kept on ice, and cultures were made from them at Plymouth 24 hours after collection. It is clear that in future attempts to make plate cultures on board a small boat in rough weather, a very delicately swung table will be necessary, or else the roll-tube culture method must be employed. Three plates on peptone agar were made from each sample, I c.c. of the sample being used for each plate. The plates were kept at the room temperature, averaging about 20° C., and the colonies were well developed after 48 hours; they appeared to be all of one kind. A count gave the following results: No. of colonies , Depth developing from in fathoms. | y¢.¢. of sample. IAW °o ° Nw ooannon~n The increase in the number of colonies at 70 and 80 fathoms is somewhat remarkable, but no conclusions in this respect can be drawn from one series of observations. The cultural characteristics of this bacterium are as follows: On peptone agar, after about 36 hours at 20° C., the colonies are white in color, circular, with a finely serrated outline and a coarsely granular appearance. Superficial colonies grow very rapidly, and may spread as a whitish semi-transparent growth of irregular shape over the surface of the agar. The deep colonies remain small, globular, and discrete. In old agar cultures a brownish tinge is developed, and the color may diffuse through the substance of the agar. On gelatin peptone growth was rapid; in stab cultures growth proceeded from the surface downwards, leaving a funnel- shaped depression of liquefied gelatin, and eventually all the gelatin became liquefied. Acid formation, as shown by the neutral-red reaction, took place in dextrose, mannite, and lzvulose, but not in cane sugar or lactose media. One thousand c.c. of Gran’s medium, inoculated on board with Io c.c. of a surface sample immediately after collection, and kept at an average temperature of about 20° C., showed the first trace of nitrite formation after 70 hours. After 84 hours a very strong nitrite reaction was obtained, and a slight ammonia reaction was given with Nessler’s reagent. The proc- ess of denitrification, even after the lapse of weeks, did not extend beyond this, and no bubbles of gas were formed. Other experiments made with subcultures from agar and gelatin media gave similar results, so that it appears that this bacterium can not entirely break down nitrates at a temperature of 20° C. The optimum temperature for denitrification pro- duced by this bacterium appears to be about 20° C., as the process was less On the Precipitation of Calcium Carbonate. 29 rapid at average temperatures of 17° C. and 25° C. At a temperature of 32° C., rapid growth took place, but no denitrification resulted. It should be noted that these temperature observations were only made with subcultures from colonies on peptone agar and peptone gelatin media, and there is reason to believe that the power of denitrification becomes diminished after cultivation on such media. Further and more accurate temperature experiments are required in which the culture medium is directly inoculated with freshly collected samples of water. This bacterium appears to be closely related to the Bacterium calcis, its chief points of difference being: 1. Lesser denitrifying power and lower temperature optimum for denitri- fication. 2. More rapid growth on gelatin media. 3. Absence of acid formation in media containing cane sugar. INVESTIGATION OF SAMPLES FROM MARQUESAS KEYS, AND EXPERIMENTAL PRECIPITATION OF CALCIUM CARBONATE BY BACTERIAL AGENCY. The Marquesas Keys constitute a coral atoll which forms part of the long chain of keys separating the Gulf of Mexico from the Straits of Florida. Within the atoll the water is very shallow and the bottom consists of a fine chalky mud many feet deep. Samples of the water from the lagoon of the atoll were sent to me at Plymouth by post, and examined 14 days after collection. On plating on peptone agar, an average of 800 colonies per I c.c. of the sample were obtained. These colonies appeared to be all of one species, and were identical in appearance and in all cultural characteristics with the Bacierium calcis previously described as occurring around the Tortugas. A suspension of these bacteria from a culture on peptone agar was made in sterile sea-water, and a similar suspension containing roughly the same number of bacteria was made from a third subculture on peptone agar of the bacteria obtained from the station 70 miles west of Ushant. I c.c. of each of these suspensions was then added to 1,000 c.c. of the modified Gran’s medium. Some of these cultures were kept at an average tem- perature of 20° C. and others at 32° C., with the following results: At 20° C. cultures from Marquesas showed trace of nitrite after 45 hours. At 20° C, cultures from Marquesas gave strong nitrite reaction after 53 hours. At 20° C. cultures from 70 miles west of Ushant showed trace of nitrite after 140 hours. At 20° C. cultures from 70 miles west of Ushant showed strong nitrite reaction after 162 hours. In both cases a slight amount of ammonia was recognizable by Nessler’s reagent when the nitrite reaction was strong, but decomposition of the nitrite did not proceed further even after 14 days. At 32° C. cultures from the Marquesas showed trace of nitrite after 18 hours. At 32° C. cultures from the Marquesas gave strong nitrite reaction after 22 hours. At 32° C. cultures from 70 miles west of Ushant never gave nitrite or ammonia reaction. The cultures from the Marquesas showed a slight amount of ammonia formation, but the decomposition of the nitrite did not proceed further. 30 Papers from the Marine Biological Laboratory at Tortugas. From these experiments it appears that the bacteria from subcultures from the Marquesas have a much greater denitrifying power than those from subcultures from a point 70 miles west of Ushant, and that as the bacteria from the Marquesas appear to be of the same species as those investigated at the Dry Tortugas, their power of causing complete denitri- fication in Gran’s medium has been lost by successive cultivations on peptone agar. The presence of the thick layers of fine chalky mud within the Marquesas Keys, and elsewhere in many places near the Florida coast, led to a con- sideration of the possibility of its precipitation by bacterial agency. Since these bacteria grow freely in Gran’s medium, the calcium salt of a simple organic acid is a sufficient source of organic food for them, and it seems probable that they would thrive in sea-water containing the products of decomposing vegetable matter, provided that the nitrate supply and conditions of light and temperature were suitable. Such conditions should be especially well fulfilled by the drainage into the sea of a well-wooded country with a calcareous subsoil, and the soluble organic calcium salts would be precipitated as calcium carbonate by the action of the bacteria. In addition, the elimination of the acid radicle from the nitrate in the process of denitrification, by whatever stages it may occur, must leave the alkaline base free to destroy the normal equilibrium of the salts in sea-water, and by increasing the alkalinity would also result in the precipitation of calcium carbonate. To test this theory cultures were made in a medium having the following composition: Caleumiea succinate: i See bec 500. ohaies cata ee eee 2.5 grams POL ASSIGN MMILEATO cbse preva sinh ys ose oe opateyesieian ac ieee 0.5 gram SOAS WALEr fon si sre ee iene Ce Oe eae eine le ese ey ota 1,000.0 c.c. Calcium succinate is soluble in these proportions and the medium is quite clear. Free growth was manifested by the cloudiness of the medium 48 hours after inoculation, and nitrite formation was apparent. After 96 hours the medium appeared quite milky and this milkiness was due to the presence of exceedingly fine particles of a substance which was soluble in dilute hydrochloric acid with evolution of gas, and was presumably calcium carbonate. In some cultures these particles settled as a definite sediment, but in others the particles were so minute that they showed little tendency to settle and could only be separated with difficulty by centrifugal- ization. The conditions determining the size of the particles formed could not be ascertained, as the size varied in cultures which were apparently made and grown under identical conditions. The addition to cultures in which the particles of calcium carbonate were so small as to remain in suspension of any foreign substance, such as finely powdered calcium sulphate or of larger particles of sand, resulted in the aggregation around them of the particles of calcium carbonate, forming a concretion around a central nucleus. These concretions were hard and of ce NE. CHANNEL Tortugas Cays ¢¢ CHART A 3 Conc epti }'San Salvador $$ $$ PS or Watling I. onl. 24 Ea hate Sa whats =iama Island. atte emegstnnas es tenes, i a BAHAMA man c .E. CHANN EY ; Tortueas -j atte eg Cays ee ° Marques as - | ol Cays . = 3 = x 7 iSan Salvador i an or Wa’ ling = 9:Conception L. a Miraporvos fi 76 aa = 100 Fathom Line. Chart Showing General Geographical Relations of the Western Bahama Island. Ae oil ¢ + es ae Nivtel 7 wrihe ig eho yt ues en BAY APSA) wile fis Scout % pally eres _ : ne odinuaiemeetaehentneneameneneneineet non = ms. Pein cham sa On the Precipitation of Calcium Carbonate. 33 but the amount of this drift was not determined, and moreover the drift caused by the wind was an unknown factor. On May 8 the wind was SSE., of force 1 at 8"30™ a.m. freshening to force 3 at 10°30" a.m. A rough esti- mate from the landfall on returning gave the drift of the boat as about 2 miles south during the 4 hours occupied in working the station; the boat had a large awning and exposed a considerable area to the wind and had drifted this distance against the wind, so it would seem that on that occasion there must have been a strong current setting south. Andros Island consists of a limestone formation, the exact nature of which has been dealt with by Dr. T. Wayland Vaughan (17). The greater part of the island is very flat and is elevated only a few feet above sea-level ; a few irregular undulations, probably not more than 100 feet high, are found, especially along the east coast. There is evidence to show that formerly the level of the land was much higher than at present, and signs of rapid erosion of the rock are everywhere obvious. One of the most remarkable features is the absence of soil, even in the well-wooded parts of the island, the trees and bushes growing directly out of crevices and holes in the rock and giving rise to practically no leaf-mold. In the numerous “ pot-holes”’ which occur all over the island, a small deposit of black leaf-mold can be found, and these ‘‘pot-holes’’ are the favorite places for the cultivation of sugar cane and bananas. ‘The erosive action of water on the rock is espe- cially noticeable where the slow drainage from an inland swamp can be traced in its course to the sea; in such a locality the hard rock is eroded, honeycombed, and undermined to a most remarkable degree, even though the amount of drainage, except after the heaviest rains, can scarcely be more than a slow trickle. Erosion of the rock along the sea-coast, where it is exposed to the action of the sea-spray, is also very marked. From the occurrence of this erosion it is obvious that all the water draining from the land into the sea must contain a high proportion of calcium salts in solution. Towards the west of the island the land is remarkably flat, and near the coast consists of white chalky mud which has partially dried and in places has formed a harder crust on the surface. These half-dried mud flats slope almost imperceptibly into the sea and are continuous with the submarine flats which extend some 60 miles off the west coast with an average depth of 2 to 3 fathoms. The mud forming these submerged flats is very soft, and near the coast it was easily possible to push a 12-foot sponge-pole down to its full length into it without touching any harder material. The surface layer of the mud for a depth of about 6 inches is of a creamy white color, but below that it is of a grayish tinge and has a slight odor of sulphureted hydrogen. Unfortunately there was no opportunity of obtaining informa- tion as to the real thickness of this layer of mud, nor of investigating more than the surface layers at any distance from the coast. Microscopical examination showed that this mud was almost entirely composed of minute unorganized particles of calcium carbonate. Near the 3 34 Papers from the Marine Biological Laboratory at Tortugas. shore a good deal of organic matter was present, chiefly in the form of de- caying mangrove roots. Farther out from the shore little organic matter was noticeable, but it was not possible to examine the deeper layers of the mud in these situations. The only organic matter that was seen consisted of the rootlets of a species of Zostera, found in occasional patches some miles off the coast. BACTERIAL INVESTIGATIONS IN THE DEEP WATER OF THE TONGUE OF THE OCEAN. Continued bad weather during the whole of our stay at Andros greatly added to the difficulties of this work, and on this account it was only found possible to work three stations. The last two were worked under the most disadvantageous conditions, the quick roll of the boat making the filling of the water-bottle with alcohol and the siphoning off of the sample under sterile conditions a matter of the greatest difficulty. The first station worked was 6 miles due east of Golding Cay, the second 14 miles east of Golding Cay, and the third 10 miles ENE.Y4E. of Golding Cay. The three stations were thus at the angles of a triangle which was nearly equilateral, the base being a little longer than the sides and running due east and west. At the first station, worked on May 8, bottom was sounded at 822 fathoms. The sea was calm at first, with a SSE. swell, but became choppy later. The wind was SSE., force 0 to I, at 8"30™ a.m., freshening to about force 3 at10"30" a.m. Thesample of the bottom obtained by the snapper- rod was of a very stiff clay-like consistency, grayish white in color, and was composed of very minute unorganized particles of calcium carbonate containing a few pteropod and globigerina shells. The following tempera- tures, to which the necessary corrections have been applied, were recorded: Depth. |Temperature. Depth. Temperature. Fms. | CG. || Fms. [0.03 Yor Siitface svtscts site sactera yee | 26.90 400... eee cree eens 18.93 | average = 8.98 TO seis alas scicle Ale cvecelebotagetete terstauets | 25.90 | 4.78 SOsa entree ee era ere 25.14 |] GOO... ......- scenes 42s | average = 4.70 HBA Aa apOOMO AOD SAIaro none aC 22.00 | 4.00 ZOO | Pe ies ere Oe | 17.13 || Bottom (822fms.).... ea average = 3.97 | . | These samples, without previous dilution, were plated in peptone agar, I c.c. of the sample being used for each plate. The agar was cooled to just under 40° C. before plating. It is very necessary that this temperature should not be exceeded, as many marine bacteria are very sensitive to heat; the use of agar at as high a temperature as 45° C. will cause the death of a large proportion of the bacteria, though in the process of plating they can be exposed to this temperature only for a very short time. The cultures were kept in the dark at the room temperature (averaging about 28° C.) and at the end of 24 hours a free growth of colonies was apparent. At the end of 48 hours the plates were counted with the following results: On the Precipitation of Calcium Carbonate. 35 Depth (in fathoms). No. of colonies developing from 1 c.c. of sample, and remarks. 822 ee atts onites ° 822 (bottom)....... 3 arses 'e I Sieley by e)\ele) sia) visiele: sls | 17 aya aieitote: ersteleteverees. 6 14 nin onl attoualie evel wens) i Is aioleretchavetetei siete et ste 16 reiclay’c ctatehevebevevors’ 1,760 | Very much overcrowded; indications of presence of many afokavelio¥ctohafele eve) ere) © 1,500 more colonies undeveloped from overcrowding. JO SOOODO DOL OES Uncountable owing to overcrowding. From these counts it is apparent that the number of bacteria falls off at some point between 200 and 400 fathoms. The second station was worked on May 11 at a point 14 miles due east of Golding Cay. The sea was calm at first, and the wind ENE., force 1, but later in the day a heavy swell set in and the wind freshened to about force 4; eventually the weather became so bad that it was impossible to work, and the station had to be abandoned before it was completed. Bottom was sounded at 890 fathoms, but there was some stray on the wire, so that the true depth was probably about 825 fathoms, as shown by the chart. The bottom consisted of fine white calcareous ooze; no remains of pteropods were seen, but some globigerina shells were present. The follow- ing temperatures were recorded: Depth. | Temperature. | Depth. | Temperature. | os sie Os oe st Fms. °C Fms. er SUMACE ed ida nti sabi sti ac 26.30 | Sis SeeWo OSE aBC nee | { Tany paverage =14.32 Pe ee re 26-49 | 400 Pees eee aoe 5b Aerob oder ny Bo ares 24.89 eich otayetrrsfehal eel ch else 9.73 = 9. TOO Nate vetetaeiea ce fei ea toate eats a telecon | 22.63 | BOI ate Metco Step caste: | 742 | Bottom............| { 4:77 average = 4.15 Samples down to a depth of 200 fathoms were diluted 1 in 100 with sterilized sea-water before plating. The following results were obtained after 48 hours’ growth: Ne: . of ens eveloping Depth. from I c.c. of sample. Fms. NOULLACE sre, ctevevsielc.e eve 16,200 LO wp aucysictarcrciacuapste @ 13,100 BOls eo seisieme veces 14,000 TOOK, feeds icrcinten foes 14,000 200 aveteeitnen cc 15,000 JOO yarsjicv cee cateseravors I4 AGE cherie te 12 The figures given represent the mean of the number of colonies develop- ing in the two plates that were made from each sample. It is here apparent that the number of bacteria per I c.c. falls off very rapidly between 200 and 300 fathoms. 36 Papers from the Marine Biological Laboratory at Tortugas. The third station was worked on May 23 at a point 10 miles ENE.W%E. of Golding Cay. The wind was east, of about force 4. As it had been blowing for the previous ten days without intermission, the sea was so rough that it was only possible to work when steaming slowly ahead into the wind. This resulted in the production of a great deal of stray on the sounding- wire, so that the number of fathoms of wire run out is greater than the actual depth at which the samples were taken; these differences will be large for the more superficial samples, but small at greater depths, as the wire strays in a curve whose gradient becomes very steep a little below the surface under these conditions. The following temperatures were recorded: Length of wire run out. T emperature. penats ae Temperature. Fms. SG: | Fms. cos ao cash tek ETSI TAN eaete, J] able Ore OF | i$.09 ;average=14.98 rool. Poona Tate UA Oars! liasees ete. 2 {T0184 j average=r0.85 At this point the station had to be abandoned, owing to the bad weather. The samples down to 160 fathoms were diluted 1 in 100 with sterilized sea- water before plating in peptone agar; the remaining two were plated undi- luted. At the end of 48 hours the following counts were made, representing the mean of the number of colonies in the two plates made from each sample: : | No. of colonies Length of wire developingfrom run out. \1c.c. of sample. Fms. | SurfaGe an lateness 15,000 BON caer eee 15,500 TOO). eects 13,700 LOO deer vcs | 13.300 25 Oe eie si aleetaveneters al 14,300 350 ~- ee. eeee | 16 The colonies developing in all the cultures were only of two kinds, the Bacterium calcis and the non-denitrifying species already described. The non-denitrifying species formed a relatively small proportion of the total, and were not found at all in cultures made from samples taken below 250 fathoms. As they appear to be comparatively inactive chemically, and as nothing is at present known concerning the part played by them in the metabolism of the sea, they will not be further considered here. A consideration of these results obtained in the Tongue of the Ocean shows that the waters down to a depth of somewhere about 300 fathoms in April 1912 contained an enormously larger number of bacteria than the water in the neighborhood of Tortugas in June 191r. The number of bacteria falls off from about 14,000 to about 12 per I c.c. between depths of 250 and 350 fathoms; the temperature at 250 fathoms was about 15° C. and at 350 fathoms about 11° C., and it was shown in June 1911 at Tortugas On the Precipitation of Calcium Carbonate. 9 that B. calcis will grow slowly at 15° C., but that growth is totally inhibited at 10°C. It would thus seem that the observed distribution of the bacteria agrees fairly with what might be expected from the temperature conditions. As regards these observations on the occurrence of bacteria in small numbers at depths below 350 fathoms, the possibility of experimental error must be considered; a leakage into the water-bottle of 0.25 c.c., as it was being hauled up through the last 300 fathoms, would account for the number found, and there are also many possible sources of error in the process of siphoning off the sample and making the cultures where a permanent labor- atory is not available. It is possible that the water below 350 fathoms was really sterile, though if so the constancy of the results obtained is curious if ascribed to experimental error. In any case, the small number of bacteria found at depths below 350 fathoms can play no part in the metabolism of the sea, since it has been shown that B. calcis is incapable of growth at the temperatures obtaining at these depths. The much greater abundance of bacteria in the surface waters of the Tongue of the Ocean than in the waters around Tortugas may perhaps be accounted for by the fact that in the Tongue of the Ocean by far the greater part of the surface water must flow over the immense chalky mud flats and shallows which bound it in most directions and, as will presently be shown, these mud flats are phenomenally rich in bacteria and are probably still being deposited by bacterial agency. HYDROGRAPHIC OBSERVATIONS IN THE TONGUE OF THE OCEAN. The samples of water taken for hydrographic investigations were analyzed by Mr. D. J. Matthews at Plymouth. With great kindness he calculated the results, and from his notes the following observations and conclusions are drawn. The samples were analyzed for salinity in comparison with the standard sea-water supplied by the Central Laboratory of the Conseil International pour |’Exploration de la Mer, and hence the results are strictly comparable with all other analyses published under the auspices of this International Council. The following results were obtained: At first station, 6 miles east of Golding Cay. Depth. |— Temp. C1 %0 | S %oo | a Cr Fms. | See Pct. (Pct. | oO | 26.90 20.06 36.24 | 20.52 23570 Io 25.90 20.46 | 36.06 | 20:71 24.57 50 25.14 20.43 36.91 | 209.66 | 24.76 100 22.00 20.28 36.64 | 29.45 25.48 200 17.13 20.08 36.27 | 20.15 26.48 400 8.08 19.58 35-37 | 28.43 27.43 600 | 4.70 19.49 35.21 | 28.30 27.90 822 3.07 19.367 34.08 | 28.11 27.79 Note.—The original surface sample was lost, owing to breakage of the bottlein transitto England. The analysis was made on a sample taken 3 days later at the same spot. In this table Cl oo means the weight of chlorine in grams found in 1,000 grams of sea-water. S 9/o means the salinity, or total weight of salt in grams found in 1,000 grams of sea-water. oo represents the specific gravity of the sample at 0° C.,and o; represents the specific gravity of the sample at the temperature ¢ at which it was collected, with no correction for pressure. 38 Papers from the Marine Biological Laboratory at Tortugas. At the second station, 13.5 miles east of Golding Cay, the following results were obtained: | | Depth. Temp. | Cl %0 | S %o0 oo a; Fms. cc \ en Bache wl weeteee | Oo 26.30 | 20.25 36.58 29.40 | 24.15 Io 26.40. | 20:33 36.73 29.52 24.24 50 24.890 || 20:305) |) 36e840 > 20.6n 24.78 r00 22/63) i) 9820-3010 nil) easO-7Gnml 20:50 25.42 200 17.42 jjpezOae2 i) 30535 29.21 26.47 300 | 14.32 | 19.81 1 35-70 28.76 26.75 400 | 9.79 | 19.56 35-34 28.40 27.27 890 | 4.15 | (No sample, bottle did not work.) | | 2] 26.3 36.58 -—-— NAUTICAL MILES oO 5 10 1S 20 SS eel Fic. 2.—Section across the Tongue of the Ocean from Golding Cay, Andros Island, showing salinities and temperatures, Owing to the uncertainty of the depths at the third station, due to the bad weather and consequent stray on the wire, it was decided not to include On the Precipitation of Calcium Carbonate. 39 these observations in a consideration of the hydrographic conditions, and to make what deductions were possible from the results obtained at the two stations given. Figure 2 shows the vertical distribution of layers of different salinities at the two stations in sphpuenie ts form. 25° TO AT eer MEOH TT UA ert TTT AUT FATHOMS Fic. 3.—Temperature curves. Continuous line =Station r. Broken line =Station 2. : Dotted line =Challenger Station No. 27.; The curves in figure 3 show the vertical distribution of temperatures at the two stations, and also give the temperatures obtained by Challenger Station No. 27 in 22° 49’ N., 65° 19’ W. March 28, 1873, where the depth was 2,960 fathoms. It is interesting to note that the actual reading at 200 fathoms at the latter station, 17.22° C., agrees more closely with the temperatures in the Tongue of the Ocean than that taken from the smoothed curve, which was 18.17° C. The curves in figure 4 show the vertical distribution of salinities at the two stations, and also the salinities obtained by the Michael Sars in 37° 12’ N., 48° 30’ W., on June 25, 1910. From these tables and diagrams it can be seen that the surface salinity increases from west to east very rapidly, 0.34 per cent in 7.5 miles, but the surface temperature is fairly uniform, between 26° C. and 27° C. At both stations the salinity increases downwards to a maximum prob- ably lying between 10 fathoms and 50 fathoms, but more rapidly at Station I, so that from 10 fathoms to 50 fathoms the salinity decreases from west to east. 40 Papers from the Marine Biological Laboratory at Tortugas. Below 100 fathoms the conditions are closely similar at both stations, as far as the observations go; the salinity decreases fairly rapidly to 400 fathoms and then more slowly to the bottom. The temperatures decrease rapidly and uniformly from the surface to about 500 fathoms, then more slowly to the bottom. 100 200 300 400 500 FATHOMS 600 700 WELLE AE Fic. 4.—Salinity curves. Continuous line =Station 1. Broken line =Station 2. Dotted line =Michael Sars Station No. 65. 800 900 There is practically no thermocline (German ‘‘sprungschicht’’) in the temperature at any depth and only a poorly marked one, confined to the upper stratum, in salinity. The absence of a temperature thermocline is remarkable (see the curves for the two stations, and the two from the Michael Sars and the German South Pole Expedition, drawn for comparison). In general, below about 250 fathoms the temperatures and salinities agree with the nearest stations of the Michael Sars in the open Atlantic; On the Precipitation of Calcium Carbonate. 4I above this depth they are higher, and differ from them and the open ocean north of the belt of calms, in the absence of a temperature thermocline, and in the maximum salinity being found below the surface. The latter points either to a considerable local supply of fresh water or to a current of lower salinity from either the Florida stream or the region of equatorial calms. Unfortunately we have no reliable salinity observations for the two latter. With regard to the accuracy of the work, Mr. Matthews makes the following remarks: THE ACCURACY OF THE OBSERVATIONS. a. Salinity.—The method of taking the samples from the water-bottle was rather incon- venient, as a siphon was used; the samples were very small, but well pre- served. The water-bottle itself might have leaked or closed at the wrong depth, as was the case with earlier models.1. That this was not so is shown by (1) The sharp fall in the number of bacteria at between 200 and 300 fathoms. (2) The close agreement of the salinities at 400 fathoms, the greatest depth at which they were taken on both stations. Station I gave 35.37; Station II, 35.34. (3) The close agreement between the bottom salinity at Station I, 34.98 at 822 fathoms, and the salinity found at the same depth at the nearest position at which we have modern observations—i. e., Michael Sars Station 65, in 37° 12’ N., 48° 30’ W., June 1910; according to the curve this is about 34.96. b. Temperatures——The National Physical Laboratory correction was given to 0.1° only, but the readings below 15° are comparable among themselves to 0.05° or possibly less. The curves of temperature for the two stations agree well in shape below 300 fathoms, but the temperature on Station II is generally slightly higher than on Station I, as a rule by an amount corresponding to a difference of depth of about 20 to 25 fathoms. Below 200 fathoms the curves for both stations agree very closely with that for Michael Sars Station 64, in 34° 44’ N., 47° 52’ W. It is almost certain from the above considerations that the results are only incorrect by the experimental errors in measuring the depth, in deter- mining the salinity (0.01° at most), and by perhaps 0.1° C. of temperature. These observations are sufficient to show that the Tongue of the Ocean is an area of considerable interest from a hydrographic point of view, and it is much to be regretted that the continued bad weather during our stay made it impossible to obtain more observations and samples. BACTERIAL INVESTIGATION OF THE CHALKY MUD FLATS WHICH ARE BEING DEPOSITED TO THE WEST OF ANDROS ISLAND. Samples of the mud were taken from the western entrance of South Bight (see Chart B), and from points 2 and 3 miles out from the shore; practically identical results were obtained from all these localities. The sample at the mouth of the bight was taken in about 4 feet of water, that 2 miles out in 7 feet, and that 3 miles out in 8 feet. The samples were neces- sarily taken from the surface of the mud. 1 The water-bottle only failed once, at about 890 fathoms at Station II. 42 Papers from the Marine Biological Laboratory at Tortugas. For bacterial examination, one part of this mud was shaken up with three parts of sterilized sea-water; this was allowed to settle for 15 minutes, and then the clearer surface layer was diluted I in 1,000,000 with sterilized sea-water. The diluted fluid was plated in peptone agar, I c.c. being used for each of the plates. The count of a number of plates after 48 hours gave 40 colonies as an average, and thus the mud itself must contain 40 X 4 X 1,000,000 = 160,000,000 bacteria per I c.c. The actual number in the mud possibly exceeds this figure, since a large proportion of the bacteria would probably settle with the larger particles after the first dilution. The bacteria found in these cultures were nearly all the B. calcis; only occasionally were a few colonies of the non-denitrifying species seen. A sample of the water taken from the surface at a spot 3 miles out from the western entrance of South Bight gave a count of 35,000 colonies per I c.c., the great majority of these being B. calcis. Subcultures of B. calcis were made in Gran’s medium, and in the calcium succinate, calcium acetate, and peptone calcium acetate media, whose composition has already been given. Denitrification in all these media was rapid and eventually complete, and was accompanied by the precipi- tation of calcium carbonate. In the last three media, which contained no solid matter and were quite clear and transparent before inoculation, this precipitation was manifested after 12 hours by the formation of a thick white cloud in the fluid, readily distinguishable from the cloudiness pro- duced merely by bacterial growth. The development of this precipitate continued rapidly during the first 48 hours, but in many cases it was com- posed of such fine particles that they showed little tendency to settle to the bottom of the flask; in other cases larger particles were formed and a sediment similar in appearance to the chalky mud of the mud flats was produced. The exact conditions determining the size of the particles precipitated could not be ascertained, as the size varied largely in cultures made at the same time, in the same media, and kept apparently under the same conditions. The addition of magnesium tartrate in small quantities (0.2 gram per 1,000 c.c.) to the culture media seemed to induce the precipi- tation of larger particles, but it did not appreciably affect the rate of growth of the bacteria. In some of the older cultures that had been kept for a week or more, the sides of the flasks were coated with a thin layer consisting of extremely minute rhombohedral crystals of calcium carbonate. Occa- sionally these crystals formed around small bubbles that had remained near the surface of the fluid, the weight of the crystals eventually caused the bubbles to sink, and then the contained gas became dissolved; in this way a number of small, hollow spheres were formed, their walls consisting of minute crystals of calcium carbonate. The formation of these curious bodies occurred especially readily in the calcium succinate medium to which 0.2 gm. of magnesium tartrate per liter had been added. The deposition of calcium carbonate in a distinctly crystalline form was only noted in old cultures, and then it was in an amount relatively extremely small when On the Precipitation of Calcium Carbonate. 43 compared to the precipitate of unorganized and amorphous calcium car- bonate. Specimens of the precipitates from some of the culture media were sent to Dr. Fred. E. Wright, of the Geophysical Laboratory of the Carnegie Institution of Washington, who described them as follows: Preparation I: Precipitate from medium composed of calcium acetate 5.0 grams, potassium nitrate 0.5 gram, peptone (Witte’s) 0.2 gram, sea-water 1,000.0 c.c., filtered and sterilized, contains two substances: (1) Fine grains of a strongly birefracting, appar- ently uniaxial, optically negative substance and with refractive index about 1.66. This is probably calcite. The grains are isolated, and no evidence of spherulitic crystallization was observed. On treatment with very dilute hydrochloric acid a noticeable evolution of carbon dioxide took place. (2) Scattered through the preparation are fine needles of a weakly birefracting substance of refractive index of about 1.525; extinction angle large. These needles are evidently selenite (hydrated calcium sulphate). Preparation II: Precipitate from medium composed of calcium succinate 2.0 grams, magnesium tartrate 0.2 gram, potassium nitrate 0.5 gram, sea-water 1,000.0 C.C., consists largely of a cryptocrystalline aggregate of a weakly birefracting substance, whose refractive index is about 1.52 to 1.53. This substance proved too fine for further determination. Scattered through this substance are rounded and irregular patches of a second crypto- crystalline substance of strong birefringence, which gives off COz when treated with dilute hydrochloric acid, and is probably calcite. Preparation III: Precipitate from a medium composed of calcium acetate 5.0 grams, sodium phosphate (Na2HPO,12H2O) 0.25 gram, potassium nitrate 0.5 gram, sea-water 1,000.0 c.c., is again very fine-grained and consists (1) in large measure of minute grains of a substance which agrees with calcite in its optical properties in so far as they could be determined. On immersion in dilute HCl a distinct evolution of COz gas was observed. (2) Of a substance whose grains are somewhat coarser than the calcite grains, their bire- fringence being medium to weak; refractive index about 1.525; biaxial and apparently optically positive; probably selenite, but not crystallized in the usual manner. The small quantity of hydrated calcium sulphate present in these precipitates is undoubtedly derived from that in solution in the sea-water with which the media were made up, but the reason of its precipitation is difficult to explain, since no such precipitation occurred in culture media kept uninoculated under similar conditions as control experiments. It would therefore appear that this deposition of calcium sulphate along with the calcium carbonate must in some indirect way be the result of bacterial action, and it would seem a possible suggestion that the odor of sulphureted hydrogen noticeable in the deeper layers of the mud flats might be due to the reduction of the calcium sulphate to a sulphide, and subsequent decom- position of the sulphide. These observations have shown that on the chalky mud flats of the Great Bahama Bank the B. calcis is found in enormous numbers, and also that this bacterium is capable of precipitating calcium carbonate from fluid media containing soluble calcium salts. It would seem a fair deduction that these mud flats, consisting of fine unorganized particles of calcium carbonate, have been precipitated by the action of the B. calcis on the soluble calcium salts carried into the sea by drainage from the land, where extensive and rapid weathering of the limestone rock is in progress. 44 Papers from the Marine Biological Laboratory at Tortugas. CONCLUSION. The observations so far available are too few, and the area they cover too small, to attempt to make any broad generalization at present. How- ever, it can be stated with a fair degree of certainty that the very extensive chalky mud flats forming the Great Bahama Bank and those which are found in places in the neighborhood of the Florida Keys are now being precipitated by the action of the Bacterium calcis on the calcium salts present in solution in sea-water. From this the suggestion is obvious that the Bacterium calcis, or other bacteria having a similar action, may have been an important factor in the formation of various chalk strata, in addi- tion to the part played by the shells of foraminifera and other organisms in the formation of these rocks. Dr. T. Wayland Vaughan has also suggested that the Miami oolite and other oolitic rocks may owe their origin to the occurrence of some diagenic change in the precipitate of very finely divided particles of calcium carbonate produced in this way by bacterial action. If this view as to the formation of chalk and oolite rocks is correct, it would seem probable that these strata must have been deposited in comparatively shallow seas whose temperature approximated to that of tropical seas at the present time. It has also been shown that bacterial denitrification is far more rapid and complete in the tropical seas around Jamaica, Tortugas, and Andrcs than in the temperate waters of the Bay of Biscay and the English Channel, and hence an explanation is provided of the relative scarcity of plankton and algal growth in the former localities, in accordance with the terms of Brandt’s hypothesis. The distribution of the bacteria, both as to numbers and species, has been shown to vary at different localities and at different depths, but there are at present too few observations to enable any conclusions or generaliza- tions to be drawn. As it now stands, the investigation can at most be considered to offer a mere indication of the part played by bacterial growth in the metabolism of the sea. To obtain a real insight into the question, it would be necessary to make more extensive bacterial and chemical observations in tropical, temperate, and arctic waters, to study the bacteriology of other areas where calcium carbonate is being precipitated from the sea, and to make further investigations in the laboratory into the chemistry of the reactions that can be brought about by various species of marine bacteria. On the Precipitation of Calcium Carbonate. 45 LITERATURE. 1. Baur, E. 1901. Ueber zwei denitrifizierende Bakterien aus der Ostsee. Wiss. Meeresunters., vol. v1. Kiel. 2. BranpbT, K. 1905. On the production and the conditions of production in the sea. Conseil Internat. Rapp. et Proc. Verb., vol. 11. ae 1904. Ueber die Bedeutung der Stickstoffverbindungen fiir die Produktion im Meere. Botan. Centralblatt, vol. Xvi. 4. Drew, G. Harotp. 1911. Report of preliminary investigations on the marine deni- trifying bacteria. Year Book No. 10, Carnegie Institution of Washing- ton. 5. ——— 1911. The action of some denitrifying bacteria in tropical and temperate seas, and the bacterial precipitation of calcium carbonate in the sea. Jour. Marine Biolog. Assoc. of the U. K., vol. 1x, No. 2. 6. ——— 1912. Report of investigations on marine bacteria carried on at Andros Island, Bahamas, British West Indies, in April 1912. Year Book No. I1, Carnegie Institution of Washington. . FEITEL, R. 1903. Beitrage zur Kenntnis denitrifizierender Meeresbakterien. Wiss. Meeresunters., vol. vil. Kiel. . FiscHer, B. 1894. Die Bakterien des Meeres. German Plankton Expedition. . Gran, H. H. 1901. Studien iiber Meeresbakterien, I. Bergens Museums, Nr. 10. Aarbog. . Kepinc, M. 1906. Weitere Untersuchungen iiber stickstoffbindende Bakterien. Wiss. Meeresunters., vol. 1x. Kiel. . KENTNER, J. 1905. Ueber das Vorkommen und Verbreitung stickstoffbindende Bakterien im Meere. Wiss. Meeresunters., vol. vit. Kiel. . Mattuews, D. J. 1913. Deep-sea_ Bacteriological Water-bottle. Jour. Marine Biolog. Assoc. of the U. K., vol. 1x, No. 4. . Murray, J., and J. Hyort. 1912. The depths of the ocean. London, . Murray, J., and R. Irvine. 1889. On_coral reefs and other carbonate of lime formations in modern seas. Proc. Royal Soc. of Edinburgh, vol. xvm. . Rasen, E. 1905. Ueber quantitative Bestimmung von Stickstoffverbindungen im Meerwasser. Wiss. Meeresunters., vol. vil. Kiel. . THOMSEN, P. 1910. Ueber das Vorkommen von Nitrobakterien im Meere. Wiss. Meeresunters., vol. x1. Kiel. . VAUGHAN, T. WAYLAND. 1912. Carnegie Institution of Washington, Year Book No. Il, pp. 153-162. ts ies dt uA af Ys lt vi past 1h avli-ruhs Sil bia ay: th +t 14 dee i mat nee “A ely yale ee be a wi fy | ‘ meee Ltt ENS ave coe Pha. 4 el ik ¥ er ih , ‘ , it ris ae - “ert, | tg \ee so oa at wo Fob vay a Te Pit ete fii ye ' is a | movie é a” yun Pal Me 4 u@, ‘ Su t ~ Vis hea De PA aif ‘ ey | U ¢ 1 y Laie Set if if HI. PRELIMINARY REMARKS ON THE GEOLOGY OF THE BAHAMAS, WITH SPECIAL REFERENCE TO THE ORIGIN OF THE BAHAMAN AND FLORIDIAN OOLITES. BY THOMAS WAYLAND VAUGHAN, Geologist in charge of Coastal Plain Investigations, U. S. Geological Survey. 47 ah AG TOOLS SHE AG. 2xA AME RAM TT OF SOMA ULAR S92" RW: nae CAA WAMAHAS FHT AO VIDEO CT, AAO PAB UAT CAA IVAW, PAMOMT? vite Saviu® lanhgotaly 2b) wradwvitr del pies info) Yo oaadts Al PRELIMINARY REMARKS ON THE GEOLOGY OF THE BAHAMAS, WITH SPECIAL REFERENCE TO THE ORIGIN OF THE BAHAMAN AND FLORIDIAN OOLITES.+ By Tuomas WAYLAND VAUGHAN. At Doctor Mayer’s request the following preliminary statement on certain aspects of Bahaman geology and on the origin of the Bahaman and Floridian oolites is submitted as a companion paper to that of Mr. Drew, whose untimely death is both a source of great sorrow to his friends and a heavy loss to science. As Drew’s investigations have proven an essential step in the solution of the problem of the origin of the extensive oolitic lime- stone formations of Florida and the Bahamas, it is fitting that an account of the processes by which they originate should accompany his posthumous memoir. Although the investigation of the rock specimens and bottom samples obtained in the Bahamas and Florida from April to July 1912 is not complete, certain definite and some tentative conclusions have been reached which may here be appropriately stated. The Bahama Islands and their accompanying shoals occupy a triangular area which lies east of Florida and north of the islands of Cuba and Haiti. The northwest corner of the triangle is in latitude 27° 35’ N., longitude 79° 2' W., about 40 miles north of Jupiter Inlet on the Florida coast. The southeast corner is Turks Islands, north of Haiti, in latitude 21° 20’ N. and longitude 71° 5’ W. The western side is a nearly north-and-south line from Little Bahama Bank to about latitude 25° 30’, whence the boundary bends into a south-of-east direction. The islands either occur on one of two large banks, the Little Bahama and the Great Bahama Banks, or they tise to the southeast of the latter bank as isolated eminences separated by water as much as 1,000 fathoms in depth. The shoalest water in the bottom of New Providence Channel between Little and Great Bahama Banks is 253 fathoms in depth; that in the Straits of Florida, between the former bank and the Florida coast, off Jupiter Inlet, is 341 fathoms deep; between the latter bank and Florida the depth is 422 fathoms. The Great Bahama Bank is separated from Cuba by the Old Bahama Channel, which ranges from 276 to 296 fathoms in depth. This bank is indented by two bodies of deep water. The first of these, theTongue Ee 1 The investigations, the results of which are here presented, were conducted under the joint auspices of the Department of Marine Biology of the Carnegie Institution of Washington and the United States Geological Survey. The former organization furnished opportunities for field work, while the latter authorized the prose- Et of the investigation as a part of my official duties, and furnished facilities for laboratory studies in ashington. 4 49 50 Papers from the Marine Biological Laboratory at Tortugas. of the Ocean, is sack-shaped and lies east of Andros Island and west of the bank, on the northern end of which is New Providence Island. Its depth ranges from 795 to 1,200 fathoms. The second indentation, Exuma Sound, is eastward from the Tongue of the Ocean, across the bank and keys ex- tending southward from New Providence Island. This sound is purse- shaped and exceeds 1,000 fathoms in depth. Water 1,000 fathoms in depth is close to the eastern shore of the Bahamas as far north as Elbow Cay on Little Bahama Bank. Eastward of the 1,000-fathom curve the bottom rapidly descends to a depth between 2,000 and 3,000 fathoms. The Bahama Islands are subaerial protuberants above the nearly level, slightly sub- merged surfaces of extensive plateaus which on one or more sides rise pre- cipitously from oceanic depths. The water from Gun Key to Northwest Passage, a distance of 67 knots, is only 7 to 12 feet in depth, while the bottom southeast of New Providence Island ranges from awash to 12 or 18 feet below water-level. New Provi- dence Island is mostly a platform from sea-level to 20 feet in elevation, with several east and west ridges standing above it; the highest of these, Nassau Ridge, rises to an altitude of about 100 feet. Andros Island has a similar physiography, a nearly level platform ranging in height from sea-level to 20 feet above that datum plane. Along its eastern front, lying near the shore, is a series of interrupted hills that range in elevation from 20 to 60, and perhaps in some instances to 100, feet. West of the coastal hills the surface is low, usually less than 10 feet in elevation, while much of the areas along the west front is affected by the tides. The Barrier Reef of Andros stands on the seaward edge of a shallow, submarine platform, at a distance of one-half mile to 2 miles off shore. The slope of the bottom seaward of the reef is comparatively rapid; between the reef and the shore the depth ranges from I to 2 fathoms. The platform between the reef and the shore is composed of hard lime rock, the surface of which is indented by numerous submarine pits. The deepest of the holes, locally known as ‘‘blue holes,” sounded was 36 feet deep. As the general aspect of these holes is similar to the abundant subaerial pot-holes, locally known as ‘‘banana holes,” formed by solution of the limestone, they are evidence of subsidence. The general country rock of the Bahamas is oolite. The sea bottom from Gun Key to Northwest Passage, New Providence Island, and Andros Island is all or mostly composed of oolite, precisely similar to that of southern Florida. The material forming the ridge at Nassau has been wind- blown, and evidently is a dune accumulation. Many or all of the ridges rising higher than 20 or 30 feet above sea-level have been formed of wind- blown material, of which oolite is an important constituent, but the oolite of most or all the lower areas is of marine origin and has not been subjected to zolian action. The finely divided calcium carbonate oozes in the shoal waters of Florida and the Bahamas have long attracted attention. Louis Agassiz, as early as 1851, noted them, as did Captain Hunt, Alexander Agassiz, and others. Preliminary Remarks on the Geology of the Bahamas. 51 Dall, so far as I am aware, was the first to suggest that ‘‘much of the limy deposit of the area behind the reefs and defended by them is probably the result of the deposition of lime originally in solution and precipitated by chemical action rather than of mechanically transported sediment.’’! Later, Samuel! Sanford became convinced that the flocculent oozes in some of the Florida bays and sounds were chemically precipitated, and so ex- pressed himself.2, The most careful study as yet made of the shoal-water bottom deposits of south Florida was the one initiated by me in 1908 and conducted in conjunction with George C. Matson. This investigation led to the conviction that the flocculent, so-called amorphous, calcium carbonate is neither of detrital nor of organic origin, and therefore must be a chemical precipitate. In 1911, G. H. Drew came to the Tortugas Laboratory for the purpose of studying denitrifying bacteria. As he discovered that ammonia was a product of the metabolism of these organ- isms he was led to conduct experiments to ascertain the effect they might have in precipitating calcium carbonate. They were found to be active in Florida; and investigations he subsequently made in the Bahamas showed 160,000,000 of these bacteria to the cubic centimeter of surface mud on the west side of Andros Island. Drew says:4 A consideration of these observations shows firstly that B. calcis is found in enormous numbers in the chalky mud flats of the Great Bahama Bank, and secondly that this bac- terium is capable of precipitating calcium carbonate from fluid media containing soluble calcium salts. It would seem a fair deduction that these mud flats have been precipitated by the action of B. calcis on the soluble calcium salts carried into the sea by drainage from the land, where extensive and rapid weathering of the limestone rock is in progress. Drew has shown what we may be confident is the major agency in pro- ducing the precipitation of the calcium carbonate ooze. Every geologist who has studied the Florida oolites, except Alexander Agassiz, has considered them marine deposits; and Louis Agassiz, Sanford, and myself considered them associated with or derived from the fine cal- careous muds. My opinion® was more definitely expressed than those of the other investigators. Rainey, as early as 1858, showed, in a remarkable book entitled ‘On the mode of formation of shells of animals, of bone, and of several other structures, by a process of molecular coalescence, demonstrated in certain artificially formed products,’ that calcium carbonate precipitated from water-soluble salts of calcium in a viscid solution form spherulites and not crystals. In 1871, Harting, in a memoir, ‘‘Recherches de morphologic synthétique sur la production artificielle de quelques formations calcaires organiques,’’® showed that calcium carbonate precipitated from a solution of a water-soluble calcium salt by the addition of the carbonate of potassium 1U. S. Geological Survey Bull. No. 84, p. 101. 1892. 2Second Annual Report Florida Geological Survey, p. 228. I910. f 1 < 3 A contribution to the geologic history of the Floridian Plateau. Carnegie Institution of Washington Pub. No. 133, pp. 114-145. I910. 4 Carnegie Institution of Washington, Year Book No. 11, p. 144. 1912. 5 Carnegie Institution of Washington Pub. No. 133, pp. 173-177. I910. 6 Verh. Kon. Akad. Wetensk., Amsterdam, vol. 13, pp. 84, 4 pls. 1871. 52 Papers from the Marine Biological Laboratory at Tortugas. or sodium is at first of gelatinous consistency and later aggregates into spherulites. In 1903, G. Linck published, in an article entitled “Die Bildung der Oolithe und Rogensteine,’’! accounts of a series of important experiments he conducted on the precipitation of calcium carbonate from sea-water by the addition of alkaline carbonates, and in 1909 another contribution from him, ‘‘Ueber die Bildung der Oolithe und Rogensteine,’’ appeared in the Jenaische Zeitschrift fiir Wissenschaft, vol. 45, pp. 267-278. According to Linck’s experiments calcium carbonate precipitated from sea-water at a temperature of 17° C. to 40° C., by adding an alkaline carbonate, is aragonite and aggregates into spherulites, the diameters of which range from 0.02 to 0.2 mm. Murray and Irvine have shown that calcium car- bonate precipitated from sea-water by an alkaline carbonate at a tempera- ture of 34° F. is calcite, at 47° F. is a mixture of calcite and aragonite, at 80° and over is aragonite.’ Drew in 1912 published the following statement regarding the calcium carbonate precipitated’ in a culture from a sample of water from Marquesas Lagoon: To such a culture a trace of very finely powdered hydrated calcium sulphate was added. This resulted in the formation of a precipitate, which, on microscopical examination, could be seen to consist of finely laminated concretions, some of which appeared to have a particle of calcium sulphate or sand as a nucleus. The concretions were soluble in dilute hydro- chloric acid with evolution of carbon dioxide. These concretions bear a resemblance to those of some oolitic limestones, and the experiment suggests the manner in which such oolites may have been formed. These references, although they lay no claim to completeness, are clear evidence that calcium carbonate, particularly aragonite, when precipitated from a water-soluble calcium salt by the addition of an alkaline carbonate, is at first of gelatinous consistency and later aggregates into spherulites.* As I did not observe oolite grains in the muds when collected in the field, I was led to the opinion that oolitization was the result of secondary changes after precipitation and directed my attention to ascertaining if such supposed changes were taking place.® Bahaman shoal-water bottom muds were collected through South Bight and off the west shore of Andros; and other samples were collected in Florida. As has been stated, these flocculent oozes when collected were not observed to contain oolite grains; however, such grains may have been present and may have escaped attention. All the Bahaman muds when examined at the end of November did contain oolite. Besides oolite grains, finely divided particles less than 0.001 mm. in diameter, some fragmental 1 Neues Jahrb. fiir Min. Beilage, Bd. 16, Dp. 495-513. 2 Proc. Roy. Sci. Edin., vol. 17, pp. 79-109. 1890. 3 Carnegie Institution of Washington, Year Book No. 10, p. 141. Io91I. 4 Without making a special diversion to go into the subject, it may be mentioned that spherulites or oolites of calcium malate and calcium phosphate are well known, and that the occurrence of spherulites of barium car- bonate and strontium carbonate have been recorded. It is therefore established that a number of salts of the calcium group of elements may assume a spherulitic form. ° Carnegie Institution of Washington, Year Book No. 11, pp. 155,157,158. 1013. Preliminary Remarks on the Geology of the Bahamas. 53 calcium carbonate, alcyonarian spicules, shoal-water foraminifera, and other micro-organisms were present." Dr. Fred. Eugene Wright has examined some of the bottom samples for me and reports that the particles large enough for ascertaining their optical properties are calcite, but the fine material can not be optically determined. A series of the muds and several powdered specimens of Pleistocene oolites from Florida and the Bahamas were tested with the cobalt nitrate reaction; and as all of these upon heating gave the purple color characteristic of aragonite, this form of calcium carbonate was shown also to be present. The muds and the Pleistocene oolites, therefore, are composed of a mixture of aragonite and calcite. The grains of the Bahama and Florida oolites range in size from 0.10 to 0.80 mm., and occasionally a grain perhaps exceeds I mm. in diameter. The spherulites or spheroid aggregates in the muds range from 0.004 or 0.006 mm. to oolite grains of ordinary size. In order to test the growth of the grains, samples of a number of muds were strained through No. 10 bolting-cloth, which has a mesh of about 0.13 mm. in diameter, and the fine material that passed through the bolting- cloth was put into bottles containing sea-water. These bottles were per- mitted to stand from about November 25 until the first week in March, over three months, when a small portion of each sample was again strained through No. 10 bolting-cloth, and the portion retained on the cloth studied. The formation of oolite grains was found to be in progress in every sample, and numerous grains had apparently grown to such a size as to preclude their passing through the mesh of bolting-cloth. The grains showed the usual forms of oolite grains: spheroids, ovoids, and ellipsoids. The larger grains had smaller diameters of 0.17 mm.; longer diameters up to 0.23 mm. Those newly formed are soft and easily crushed by any kind of pressure. The experiments indicate increase both in number and in size of grains. The precipitated calcium carbonate may segregate around a variety of nuclei, for instance, spherulites or round aggregates formed of the precipi- tated material, small grains of sands, shells of foraminifera, and gas bubbles.’ Although there is need for additional study of the factors that accelerate, retard, or inhibit the formation of spherulites and initial round aggregates and the subsequent growth of the grains, the empirical facts in the process of the formation of the Floridian and Bahaman oolites are clear. They are as follows: 1. Denitrifying bacteria are very active in the shoal waters of both regions and are precipitating enormous quantities of calcium carbonate which is largely aragonite. 2. This chemically precipitated calcium carbonate may form spherulites or small balls which by accretion may become oolite grains of the usual size, or it may accumulate around a variety of nuclei to build such grains. 1 The investigation of these muds is stillin progress {June 1913] and will not be finished for some months, The completed results will appear in a subsequent publication. 2 See the papers by Linck, Drew, and Vaughan, already cited. 54 Papers from the Marine Biological Laboratory at Tortugas. The deduction may be made that all marine oolites, originally composed of calcium carbonate, of whatever geologic age, may confidently be attrib- uted to this process. Two other important deductions may be made from the knowledge of this process, viz: 1. Neither the Bahamas nor the oolitic keys of southern Florida are coral islands, but they have been formed by this other process. Elevated coral rock is exceedingly scarce in the Bahamas and the Recent reef of Andros is comparatively insignificant as a constructional geologic agent. The material composing the land masses and much or most of the submarine platforms of the Bahamas is thus removed from the category of ‘‘coral rock’”’ and the living reef reduced to a subordinate ratio as a builder of limestone. 2. Drew’s studies (unfortunately incomplete) of the distribution of denitrifying bacteria have shown them to be most prevalent in the shoal waters of the tropics. They therefore conform to the temperature relations enunciated by Murray! for the distribution of lime-secreting organisms. By combining the results of Drew and Murray, the deduction seems warranted that great limestone formations, whether they be composed of organic or of chemically precipitated calcium carbonate, were laid down in waters of which at least the surface temperatures were warm, if not actually tropical. This brief paper may be closed with the following statement and tenta- tive outline of the geologic history of the Bahamas, with special reference to the eastern part of Andros Island: The presence of oolite containing Pleistocene marine fossils above sea- level indicates elevation in the Bahamas; the presence of submerged solution pot-holes, not filled with sediment, indicates that the last movement was downward. TENTATIVE OUTLINE OF THE PLEISTOCENE AND RECENT GEOLOGIC HISTORY OF THE BAHAMAS, AND ESPECIALLY OF ANDROS ISLAND. (1) During Pleistocene, perhaps earlier Pleistocene time, numerous shal- low submarine banks existed, on which calcium carbonate was chemically precipitated mostly as aragonite, and this was converted into oolite. (2) This submergence was followed by uplift with the production by marine erosion of a platform and a margining shore cliff, and the formation of shore dunes paralleling the windward sea-front. (3) Apparently there was further uplift and pot-holes were subaerially formed by solution on the platform between the dunes and the sea-front. At this time the land on the east side of Andros Island stood about 40 feet higher than now. (4) The last event was a subsidence of about 40 feet, admitting the sea back to the edge of the previously cut low scarp, and the present barrier reef developed on the off-shore margin of the submerged platform. 1 Natural Science, vol. 2, pp. 25-27. 1897. THE BUILDING OF THE MARQUESAS AND TORTUGAS ATOLES*AND A SKETCHY OF: THE ‘GEOLOGIC HISTORY*OF THE FEORIDA REEF: TRACY. BY THOMAS WAYLAND VAUGHAN, Geologist in charge of Coastal Plain Investigations, U. S. Geological Survey, 55 IV. THE BUILDING OF THE MARQUESAS AND TORTUGAS ATOLLS AND A SKETCH OF THE GEOLOGIC HISTORY OF THE FLORIDA REEF TRACT. By THomsas WAYLAND VAUGHAN. INTRODUCTION. The following brief article is supplementary to my paper ‘‘A contri- bution to the geologic history of the Floridian Plateau,’”! in which it is stated that ‘‘the geologic history of the Marquesas and Tortugas is reserved for future consideration.’”’ The study of the geology and the geologic processes of the Florida reef tract, and especially of the Tortugas and the Marquesas, has been continued since 1910. During the field season of 1913 Mr. R. B. Dole, of che U. S. Geological Survey, undertook a special investigation of the solvent effect, by virtue of the content of carbon dioxide, of sea-water flowing into and out of Tortugas Lagoon. As the various lines of investigation have given definite results, the principal factors in the formation of the Marquesas and Tortugas atolls and their inclosed lagoons now seem clear, although some details still require elucidation. It is now also possible to outline with a feeling of confidence the salient geologic episodes in the history of the entire Florida reef tract and to institute comparisons with other coral-reef areas. As this paper is only a preliminary statement, it is intended subsequently to publish a more comprehensive discussion of the different problems here treated in a cursory manner. BUILDING OF THE MARQUESAS AND TORTUGAS ATOLLS. An inspection of the charts of the U. S. Coast and Geodetic Survey, No. 1252 for the Marquesas and No. 471a for the Tortugas, will show a striking similarity in the configuration of both, for in each banks or keys surround a central depression and in each there are major entrances in the southeast, southwest, and northwest quadrants. The Tortugas, however, are of a more elliptical form than the Marquesas, the axis of the ellipse extend- ing from northeast to southwest. The Marquesas lagoon is only I to 15 feet in depth, while that of the Tortugas is from 7 to 13 fathoms. In both the principal key or bank slightly under water extends from the southeast 1 Carnegie Institution of Washington, Pub. 133, pp. 99-185, 15 pls. rgr0. 57 58 Papers from the Marine Biological Laboratory at Tortugas. to the northwest entrance, or is on the east, northeast, and north sides. The keys and banks between the southeast and southwest passages and between the southwest and northwest passages are minor as compared with the larger one mentioned. It may also be pointed out that the Marquesas occur just west of Boca Grande Channel, and thai the Tortugas lie just west of Rebecca Channel. That the Marquesas were formed through the agency of waves and currents operating under proper conditions has been evident for some time, but the processes by which the Tortugas were built were less clear. It was thought possible that submarine solution had been efficacious in the latter, while it was apparent that it was ineffective in the former. In order to answer the question for the Tortugas, two lines of inquiry were undertaken. One of these was the examination of the bottom of the Tortugas Lagoon, to discover whether or not sediment is there accumulating; the other was to ascertain by chemical examination whether or not there was any excess of carbon dioxide in the water flowing into the lagoon and whether there was any difference in the quantity of carbonates in the outflowing and inflowing water. The latter investigation was undertaken by Mr. R. B. Dole. The bottom samples collected at numerous localities in the Tortugas established that the bottom in the channel between Garden Key and Bush Key, in Bird Key Harbor, and in the lagoon channels west and east of White Shoal and north of that shoal is formed of soft calcareous mud, largely of the kind Drew showed to be precipitated by denitrifying bacteria. The examination showed conclusively that sedimentation predominates over the removal of material, perhaps except where currents may corrade the sides or bottoms of the channels along which they flow. The conditions in the Tortugas lagoon are similar to those in the bays and sounds behind the Florida keys and to those in Marquesas Lagoon, in all of which filling by deposition is progressing. The specimens examined by Mr. Dole were collected twice a day on the principal flowing and ebbing tides for nearly a lunar period, from May 20 to June 15, near buoy No. C3, in Southwest Channel, off the southern end of White Shoal. Mr. Dole’s results are summarized on page 74 of this publication. They show that none of the water, neither that flowing into nor that flowing out of the lagoon, contains any dissolved free carbon dioxide, that all the carbon dioxide in the water is present in the carbonate or bicarbonate form, and that the inflowing water possesses no power to dissolve calcium carbonate by virtue of its content of carbon dioxide, as its capacity for the solution of lime has been exhausted before flowing into the lagoon. The chemical investigations of the waters and the study of the bottom deposits gave mutually confirmatory results, that the water flowing through the Tortugas Lagoon does not dissolve calcium carbonate and that the lagoon has not been formed by solution. As two kinds of phenomena observed in the Tortugas might be con- sidered to afford evidence in favor of solution, both will be briefly discussed. Building of the Marquesas and Tortugas Aliolls, etc. 59 One is the frequent occurrence of submerged coral heads, the interiors of which have been removed, leaving a partially inclosing skeletal shell covered by living polyps. Some of these heads have a suggestive resemblance to atolls, but the resemblance is only superficial. The excavation of the interior is 10 be explained by the upper surface of the envelope of living tissue having been damaged in some way, thus giving an opportunity for boring organisms of great variety to begin and prosecute their work of disintegrating the skeleton. Waves and currents wash out the broken-up material, forming a cavity which is bounded by a shell of substance whose outer surface is covered by living polyps. The other phenomena that might be attributed to solution are the overhanging projections along the steep, submarine cliffs or rock pinnacles which in places occur on the sides of channels. Since these stand against the sides of the currents it is probable that submarine scour, corrasion, has been a factor in producing them; and subaerial modification deserves consideration, as will later appear. As the dynamic agencies have been more and more thoroughly studied, the necessity for referring any phenomena observed in the Tortugas to sub- marine solution has been eliminated, aad the conclusion definitely reached that such solution produces no geologic effects of importance. As the production of the Marquesas and Tortugas atolls with their inclosed lagoons by solution is eliminated, other factors, especially waves and currents, will be considered. There are two current-shaped physio- graphic forms to which it is desired particularly to call attention—these are current-shaped crescents and current-shaped linear ridges. With reference to crescents it may be said that if an obstruction lies across the line of direction of a constant current, the current shears to each side and will drift detrital material in such directions as to form a crescentic accumulation, the bow of the crescent facing the current, while the horns of the crescent will curve before the current. An eddy may be produced in the space between the horns of the crescent, and a deposit may be formed there.! Crescentic sand-dunes and crescentic keys are both well known. Should there be no cross obstruction, currents both aerial and aqueous will form linear ridges, under conditions which induce dropping material. Both linear sand-dunes and linear submarine banks are well known. Of current- transported detritus, crescentic accumulations lie across the main direction of the current, while linear ridges lie along its direction. The currents of the Florida keys are of three kinds: wind-formed currents, which are accompanied by waves; the Florida counter current; and tidal currents. The winds at Key West, where accurate records have been kept, prevailingly range in direction from northeast to southeast, those from the east predominating over those from any other direction. Both the Marquesas and the Tortugas lie within the tract of the Florida counter current, which moves in general toward the west. The directions of the 1 Dr. A. G. Mayer called my attention to the occurrence of this eddy. 60 Papers from the Marine Biological Laboratory at Tortugas. wind-formed currents and the counter current are concordant and they therefore cooperate. The tidal impulse, however, is roughly from north to south and the direction of tidal currents is therefore transverse to the other currents. The principal arcs of the rims of both the Marquesas and the Tortugas, 1. e., those from the southeast to the northwest entrance, are bowed toward the east, against the prevailing direction of the wind and that of the counter current, while the northern limb of the arc in each instance trails with these currents. These arcs therefore satisfy the requirements of wave and current-shaped arcs. The southern sides of both the Marquesas and Tortugas are broken by southeast and southwest passages. These breaks are probably to be attributed to cross-tidal currents. Also both the Marquesas and Tortugas have a northwest passage. The explanation of this is probably as follows: In the Tortugas, although the data on currents are inexact, the available evidence indicates a stronger tidal inflow than outflow, much of the water of the receding tides not passing through the lagoon but around the outside of its rim. The water coming into the lagoon would tend to be driven toward the west by the winds, and would consequently find its outlet in the northwest quadrant. In the Marquesas, the land area west of the southeast passage follows a line that would be expected of the southeast horn of a crescent formed under conditions of wind and currents known to prevail there. The formation of the bank and the small keys on it on the west side of the lagoon is more obscure. Perhaps they were formed by winds and accom- panying waves that come, especially during hurricanes, from the west and by a current eddy produced to the leeward of the bow of the crescent. The main agencies whereby the principal arc of the rim of the Tortugas was formed seem definitely known, but those building other parts of the rim are still somewhat vague. Between what is known as Five-foot Channel and Southwest Passage are holes, some of which are 6 fathoms or more in depth. It is evident from the distribution of these holes that once there were several channels trending more or less from southwest to northeast and approximately following the course of tidal flow, the channels bounded by banks elongate in the same direction. Detritus carried westward by waves and currents has filled parts of the channels and broken them up into holes, some series of which may still be recognized. Loggerhead Key, as well as the bank on which it stands, is elongate from southwest to northeast and has each of its ends somewhat curved toward the west. The general direction of the bank is approximately along that of tidal flow, but it is not an unmodified linear ridge coinciding in direction with that of one current, for it is evident that the prevailing run of the waves before the wind, and perhaps the counter current acting at its southern end, have curved its ends westward. The effect of waves running before the wind in shaping a key was studied in detail on Logger- Building of the Marquesas and Tortugas Atolls, etc. 61 head Key, where both the north and south horns of the key curve before the prevailing easterly winds. A reversal of a part of the horn of the north end of the key was observed during a swell from the northwest. During this reversal of the direction of the run of the waves, a reversed crescent was formed at the end of the spit, the bow of the crescent being toward the current while the horns curved before it. The more indurated beach rock on each side of Loggerhead Key slopes toward the sea, showing that the key is wave-built, on the surface at least. In this connection it may be said that a current too weak to pick up and carry detritus will, when agitation has lifted particles above the bottom, appreciably move them in the direction of the current. Loggerhead Key stands on the lee side of the lagoon, where waves from the east would be weaker than on the windward side, and where waves from the west would exert their full force. The following is offered as an hypothesis for the formation of the bank on which this key stands: It is largely or mostly a detrital ridge, formed in a partial eddy on the leeward side of the lagoon, mainly along the direction of flow of the stronger tidal currents. Wave agitation from the east and west has facilitated the movements of detritus and has built the sloping beach rock on both the lagoon and the sea sides. The stronger winds from the east have given a slight westward curvature to the extremities of the bank. Before leaving the discussion of Loggerhead Bank and Key, attention will be directed to changes that have occurred since October 1910, when the last severe hurricane visited this region. Since that time the waves have acted with only slight cessation on the eastern side of the key and have cut away the eastern side of its northern end, while building has taken place on the western side; thus the outline of the northern end of the key has shifted westward. Accurate measurements of some of these changes have been kept but will not be given here. Besides the banks and keys forming the perimeter of the Tortugas, there are also shoals and some small keys in the lagoon. Some of the shoals, White Shoal for example, lie along the direction of flow of the major tidal currents, although with their ends slightly curving toward the west; others are isolated rocks. That some of the latter may be coral masses is suggested by an experience on Garden Key, where all large rocks examined proved to be dead coral heads. The rocks composing both Marquesas and Tortugas are known only from superficial examination. The foundation of che Marquesas may be oolite, similar to that found on Boca Grande Key, as some of the bottom samples from them were found to contain hard oolite grains. The keys, however, are composed of calcareous detritus, of which Halimeda is an im- portant constituent. There is no important coral growth around the Marquesas. The keys of the Tortugas are mostly composed of the com- minuted remains of a considerable variety of organisms that secrete calcium carbonate, mollusca, corals, nullipores, echinoids, etc. The corals here, in contradistinction to the Marquesas, are important. Coral reefs, patches, 62 Papers from the Marine Biological Laboratory at Tortugas. or heads occur along the west side of Loggerhead bank throughout its entire length and around both its southern and northernends. Bush Key Reef, southwest of Southeast Passage, is luxuriant, and outer reef patches occur northward of Southeast Passage. Reef corals also occur at many places along the sides of channels within the lagoon. Other species of corals, not so large and massive as those on the reefs proper, live on the shallow flats, where not overwhelmed with detritus. Although it has not been practicable to make for the Tortugas a quantitative estimate of the pro- portions of material contributed by the different kinds of organisms, corals are rather surely the most important. The Marquesas, therefore, are composed of calcareous detritus mostly of other than coral origin; while the Tortugas are composed of calcareous material, of complex origin but largely due to the activity of corals. The foregoing remarks may be summarized as follows: (1) Submarine solution has not been instrumental in the formation of the Marquesas or the Tortugas lagoon; (2) the atoll rings are constructional phenomena and were shaped by prevailing currents, those caused by winds, the Florida counter current, and tidal currents. The detrital material on which these agencies have worked in the Marquesas is mostly not of coral origin; while in the Tortugas, although of complex composition,! it is largely composed of coral débris. The relative ages of the Marquesas and Tortugas atolls and their relations to oscillation of water-level will now be briefly considered, and in this connection further attention will be given to the probable relations of corals to the formation of the latter atoll. The Marquesas rim is composed of unconsolidated detrital material and evidently requires no oscillation of water-level for its formation. No indication of change of level during or since the building of the rim around the lagoon was observed. Such a structure as the Tortugas would scarcely be built in the Florida region through material derived from reef corals without change in depth, as the depth of water in the lagoon, 13 fathoms, and outside the atoll rim, 13 fathoms or more, is greater than that in which the Florida reef species are known to flourish.2 Other evidence, to be adduced in giving an account of the coastal oscillacion, renders it reasonably certain that the Tortugas were initially outlined during subsidence.’ Certain facts indicate that the history of the Tortugas is more complicated than that of the Marquesas. 1 The following literature is cited in this connection: Vaughan Cornish. On the formation of sand dunes. Geog. Jour. vol. 9, pp. 278-309. 1897. C. Hedley and T. Griffith-Taylor. Corals of the Great Barrier, Queensland, a study of their structure, ig lis ae and relation to mainland physiography. Austral. Assoc. Adv. Sci. Adelaide meeting, 17 pp., 3 pls. 1907. Ev bees ones. Coral and atolls. London. Lovell Reeve & Co. Ltd. (especially chapters x11 and XIII, PP. 253-277). I910. James Bryce. South America observations and impressions. New York, Macmillan Co. 1912. [Pp. 58, 59 give a good description of crescentic sand-dunes, ‘‘the convex of the crescent always facing the wind,” on the great inner plateau of Peru.] 2 In the following discussion of oscillation the terms ‘‘uplift,’’ ‘‘depression,’’ and ‘‘subsidence’’ are used with reference to land, but it is recognized that oscillation may be due to negative or positive movement of sea-level independent of crustal movement. : ’ The wells bored on Key Vaca by the Florida East Coast Railway, under the supervision of Samuel Sanford, showed the thickness of the elevated coral reef rock to be 105 feet (= 173 fathoms), indicating that the Pleistocene barrier reef was formed under conditions of subsidence, similar to those postulated for the initial building of the Tortugas rim. Building of the Marquesas and Tortugas Atolls, etc. 63 Most or all of the material above sea-level in the Tortugas is unconsolidated, except an outer crust of some of the beach rock, as observed on Loggerhead Key. Extending beneath the sea-level, however, to a depth of 8 feet or more, is beach rock that appears more indurated. Harder rock was also observed at a depth of 3 or 4 feet under water on Southwest and Northeast keys, the subaerial portions of both have been cut away by wave action, and in places submarine cliffs of hard rock, which has been undercut and overhangs the sides of the channels, are evident at depths of over 20 feet. The presence of an older formation for the living reef corals is clear. It seems probable that this rock was subaerially indurated and then depressed. The probability of this subsidence will be further discussed in a succeeding paragraph, wherein the later movements of the Florida coast line will be more fully considered. It appears that the Tortugas atoll with its inclosed lagoon was outlined previous to the present development of the coral reefs of that area, that it was elevated about 50 feet, and that there has been a subsequent (the latest) depression; the land after the oscillation perhaps lacked 10 or 15 feet of being depressed to its former level, thus leaving the keys standing somewhat higher than before. It has already been stated that the skeletal remains of corals are not only an important constituent of the Tortugas rocks, but are rather surely the most important contributor to its composition. Whether or not the various keys, banks, and isolated rocks were outlined by coral reefs may not be ascertained, but as coral reefs obey the laws of current-moved detritus, in many instances being the principal single source of the detritus, a positive answer to this question is not essential for the validity of the explanation here presented for the building of the Tortugas. Hedley and Taylor say in their paper on the Great Barrier, referred to in the footnote on page 62: The growth of an individual reef is shown to proceed in a regular cycle. If the reef reaches the surface with its axis along the wind, then its shape endures; but if across the wind, then its extremities are produced backward, forming first a crescent, later a horseshoe, and lastly an oval, thus inclosing a lagoon. SKETCH OF THE GEOLOGIC HISTORY OF THE FLORIDA REEF TRACT. The probability of elevation and subsidence in the Tortugas area leads to a consideration of evidence from which the later movements of the Florida coast line have been inferred and of conditions antecedent to the development of the living Florida barrier reef. That the last important movement of the Florida east coast was downward from Jacksonville to San Augustine is attested by the submerged channel of the mouth of St. Johns River and the submarine fresh-water springs off San Augustine. Late depression of the west coast is positively shown not only by the forms of Tampa Bay and Charlotte Harbor, but also by submerged channels off shore in that region. As the channels off this coast, however, appear not to affect the 10-fathom curve the depression was probably not so great as. 60 feet. The very ragged character of the coast line from Cape Romano to 64 Papers from the Marine Biological Laboratory at Tortugas. the head of Florida Bay, with its salients, reentrants, and almost numberless small islands lying near shore, as Ten Thousand Islands, separated by an interlocking maze of channels, suggests that the southern end of the penin- sula to its very tip has undergone subsidence at no distant date. On keys west of Bahia Honda Sanford noted holes, which, at a distance of a quarter of a mile or more from shore, connect with underground passages containing salt water; and on more easterly keys formed of elevated coral reef rock he observed free openings or cavities that extend to a depth as great as 30 feet below sea-level. The evidence is clear that the keys participated in the uplift and subsequent depression that affected the mainland and that at one time they stood more than 30 feet higher with reference to sea-level than they now do. This uplift and the subsequent depression, according to all available evidence, extended to the Tortugas. The highest points to which the elevated reef extends are on Windleys Island and Plantation Key, where elevations of 18 feet are attained. Sup- posing these points to have been at sea-level, an uplift with reference to sea-level, of somewhat more than 48 feet, must have taken place since the elevated reef was formed. The evidence presented shows that the platform, on the outer edge of which the present barrier reef of Florida is principally growing, has geo- logically, just antecedent to its present relations to sea-level, stood 30 feet or more higher. It is on this shelf, the last oscillatory movement of which was one of depression, that the living reefs of the Florida barrier have established themselves and have grown. However, although the last earth movement has been downward, the net result of the last oscillation, 7. e., uplift followed by subsidence, has been to bring the surface of the sub- marine shelf nearer to the surface of the sea. Although the building of the platform seaward of the Pleistocene barrier reef needs further study, it is known that the base of the Pleistocene reef is about 100 feet below sea- level. As the reef would not have begun to grow in a marked depression, it is probable that the water seaward of it was not shallower than that at its base. The maximum depth of Hawk Channel is about 7 fathoms, while in places the living barrier reaches the surface of the sea. Therefore, since the initiation of the Pleistocene reef the bottom seaward of it has been built up to a thickness ranging from 70 to 100 feet. As the question, how much of this material may be Pleistocene, can not now be answered, it is not known whether or not the living reef has Pleistocene corals beneath it. However these questions may ultimately be answered, it is clear that both addition of material (upbuilding) and uplift have contributed to raising the surface of the basement of the Recent reef, notwithstanding that the last oscillatory movement carried the land surface downward. The hydro- graphic charts (see particularly U. S. Coast and Geodetic Chart No. 165, Hillsboro Inlet and Fowey Rocks) clearly show the northward extension of the reef platform beyond the limits of the living reefs, which are merely superimposed on a platform whose existence antedates their own. Building of the Marquesas and Tortugas Atolls, etc. 65 The conditions under which the Pleistocene barrier reef was formed may be reconstructed by using the results obtained from the recent geologic investigations on the Florida mainland and the records of the wells bored by the Florida East Coast Railway on Key Vaca. Pliocene deposition was followed by uplift, the ensuing Pleistocene deposits being laid down on the eroded surface of those preceding. The Pleistocene reef, which is 105 feet thick, was therefore formed on a subsiding sea-bottom succeeding the uplift at the end of Pliocene time. The oscillations of southern Florida with reference to coral reefs may be tabularly expressed as follows: Oscillations of Southern Florida. IRECONES, «scat Depression (modern) reefs. Uplift. Depression (Pleistocene reefs, parts of which now stand as Pleistocene... .... much as 18 feet above sea-level). Uplift. Phocenes.-asete Depression (some reef corals but no well-developed reefs). The results of the recent investigations by Sanford, Drew, Dole, and myself have revealed the salient facts in the geologic history of the tract of the Florida keys and reefs, except the immediate foundation rock of the living barrier reef. Great progress was made by clearly showing the process by which the oolites were formed and by proving that the atoll rims of the Marquesas and Tortugas are not due to the solution of an interior mass of limestone but to constructional geologic processes. The hypoth- esis that Biscayne Bay was formed by marine solution is also definitely disproven. This history may be summarized as follows: Pliocene deposition was followed by uplift, which was succeeded by depression; during this Pleisto- cene subsidence along a curve from the eastern side of Biscayne Bay, first trending southward and then bending westward, a barrier coral reef flour- ished, separated by a channel from the main bank on which the Miami oolite was forming or had formed in strongly agitated waters. West of the coral reef, on an extensive flat in shoal water, the Key West oolite was formed, while still farther to the westward the Tortugas were out- lined under the influence of winds and currents. This period of events was succeeded by the elevation of the entire key region to more than 50 feet above its previous level. This uplift was succeeded by one of depres- sion, lowering the surface 30 feet or more, establishing practically the same relation of the sea-level to land that now prevails. Subsequent to the beginning of this last depression the present barrier reef has developed seaward of the keys on a platform already prepared for it, the Marquesas have been formed by winds and currents, and coral reefs have reestab- lished themselves in the Tortugas. 5 66 Papers from the Marine Biological Laboratory at Tortugas. COMPARISONS OF THE FLORIDA REEF TRACT WITH SOME OTHER CORAL REEF AREAS. In order to compare the relations of the barrier reef of Florida with the last change of sea-level in Cuba and in Andros Island, Bahamas, the following statements will be made: Hayes, Vaughan, and Spencer! have shown, as is evidenced by the pouch-shaped harbors of the Cuban coast and filled channels, such as the submerged filled channel in Havana Harbor, that the last movement of the Cuban coast has been downward with refer- ence to sea-level. As an account of the Cuban reefs would be out of place here, reference is made to that published by A. Agassiz.2, The platform on which the Cuban reefs grow has been brought to its present position by subsidence. The great barrier reef of Andros Island, Bahamas, as I have shown,’ occupies the outer edge of a depressed platform. Therefore the Floridian, Andros, and Cuban reefs all have a similar relation to the oscilla- tion of sea-level, as in each instance there has been elevation antecedent to depression which has brought the platforms on which the reefs stand into their present positions. There is, however, in one important respect, a difference between the relations of the Recent reefs of Florida, Andros Island, Bahamas, and of those of Cuba to Pleistocene oscillation. The oscillations in Florida and Andros Island have taken place without notable differential crustal move- ments; while in Cuba there was notable deformation accompanying the oscillations. The Pleistocene terraces rise in height toward the eastern end of Cuba in Oriente Province, where altitudes of about 600 feet are attained near Cape Maisi; while in Santa Clara, Matanzas, and Havana provinces terraces on the north side are distinct and rise in several steps to heights of about 200 feet, but are absent or indistinct on the southern side.’ These facts show that there has been pronounced tilting in late Pleistocene time. In Barbados Pleistocene reefs extend to 1,000 feet in elevation. By working out the salient features in the development of the Florida reefs and instituting comparisons with the West Indies a basis has been supplied for explaining certain puzzling relations of the reefs in the Tropical Pacific. A. Agassiz discovered that in the Paumotuan atolls the Recent corals were growing as a thin crust on an older limestone foundation. His explanation of the formation of the atolls by the destruction of the interior of a limestone mass must be discarded, as they were certainly not formed in that manner but by constructional agencies. There was evidently a period of atoll formation in the Paumotus previous to the growth of the Recent corals, which have established themselves on an atoll basement already prepared for them. A great development of Pleistocene (and per- haps late Tertiary) coral reefs in the tropical Pacific has been proven in the most convincing way. .. } Report on a geological reconnaissance of Cuba, made under the direction of General Leonard Wood, Military Governor, 1901. Report of the Military Governor for 1901. 2 Bull. Mus. Comp. Zool., vol. 26, pp. 133-136. 1894. 8 Carn. Inst. Wash. Year Book, No. 11, p. 154, 1912; Carn. Inst. Wash. Pub. 182, page 50, 1913. ‘ Hayes, Vaughan, and Spencer, Geological reconnaissance of Cuba, pp. 18-20. Building of the Marquesas and Tortugas Atolls, etc. 67 There is abundant evidence of differential crustal movement in the tropi- cal Pacific similar to that indicated for the West Indies. E. C. Andrews! and C. Elschner? have both described warping or tilting, the former for the Fijis, the latter for the Pacific more or less in general. Agassiz described an elevated atoll, Makatea, in the Paumotus, and Andrews has given a detailed description of the elevated atoll of Mango, Fijis. The old reefs have been subjected to differentiated earth movement, so that in certain places old atolls, such as Rangiroa and others in the Paumotus, now stand at or near sea-level, while other atolls have been uplifted to heights as great as 230 feet in Makatea and 600 feet in Tuvutha, Fijis. In other areas there has been depression, Bora-Bora, for instance, as Dana, P. Marshall, and more recently W. M. Davis, have shown. In conclusion it will be said regarding the Pacific coral reefs: (1) Atolls are not formed by solution, but by constructional geologic agencies; (2) there was in the tropical Pacific a great development of Pleistocene and perhaps late Tertiary reefs which have subsequent to their formation been sub- jected to extensive differential crustal movement; (3) these deformed older reefs are frequently the basement of the Recent reefs. Although important contributions have already been made to the study of Pleistocene and Recent oscillation and deformation in the Tropical Pacific, there is great need for more extensive and more detailed investigations of this phase of the coral-reef problem in order to ascertain more accurately the relations of the older to the Recent reef series. 1 Notes on the limestones and general geology of the Fiji Islands; Bull. Mus. Comp. Zool., vol. 38, pp. 27-30. 1900. Wee Corallogene Posphat-Inseln Austral-Oceaniens, pp. 15-10, Liibeck. 1913. ry VAe i my nen ret V4 ow MIE iat vy Oral PT ee Hig opt nae aa i Aven; Tea aa Ub’ ral hire is TASHA ur Mes fiw be aay Lig ” AS etal wih’ ry rt its . wiigia. a { ere 4 | * be aie ld Jd ere ee eee AO pe ae; a. VE SOME CHEMICAL CHARACTERISTICS OF SEA-WATER AT TORTUGAS AND AROUND BISCAYNE BAY, FLORIDA. BY RK: B. DOLE; Chemist U. S. Geological Survey. One Map. 69 HAVAW 032: TO Z3eSTSAAeY tunes | EMyAQ2IG CMUGRA OVA BAOPMOT Ta air, AOS YAS a “ a = a - Re tO a ak ie hele ton AT whee sig ort 5 ~ 4 i i ‘ ' wy i : Pr iS z - Tere he Port ee SOME CHEMICAL CHARACTERISTICS OF SEA-WATER AT TORTUGAS AND AROUND BISCAYNE BAY, FLORIDA. BYe ke be DOLE. INTRODUCTION. The chemical tests at Tortugas were performed by the writer in June 1913, in the Marine Biological Laboratory, Tortugas, Florida, for the primary purpose of ascertaining what soluble effect, if any, carbon dioxide in sea-water might have on coral and other deposits of calcium carbonate. The tests of waters from Biscayne Bay were made to ascertain the differ- ences in concentration of sea-water in the bay and the diluting effect of Miami River. The samples for the latter work were collected by the writer June 23, 1913, and they were examined during the summer by E. C. Bain, junior chemist, U. S. Geological Survey. DETERMINATION OF CARBON DIOXIDE. To 100 c.c. of the sample, in a Nessler tube measuring 17.5 cm. to the graduating mark, 10 drops of I per cent phenolphthalein was added and the solution was titrated to an acid reaction by means of N/20 sulphuric acid. This is the usual procedure for estimation of normal carbonates in water. The result of the titration has been calculated to ‘carbon dioxide (CO:) present as carbonate (CO;)” by means of the formula A = 22Q/d, in which A represents milligrams per liter of carbon dioxide, Q the number of cubic centimeters of N/20 sulphuric acid required, and d the density of the water. A sufficient excess of N/20 sulphuric acid was added to 200 c.c. of the sample in a Jena flask and the solution was gently boiled long enough to drive off carbon dioxide, after which the total quantity of acid consumed was determined by titrating the excess with barium hydrate in presence of phenolphthalein. This procedure was followed by Fox! and others in determining the alkalinity of sea-water. The ‘‘carbon dioxide (COz) present as bicarbonate (HCO;)’’ was computed by means of the formula B=11(Q’—4Q)/d, in which B represents milligrams per liter of carbon diox- ide, and Q’ the total quantity of acid required. The result of the second titration has also been expressed as “total alkalinity in equivalent OH”’ in order to afford comparison with the results of certain other analysts. This calculation has been made by the formula C = 4.275Q’/d, in which C represents the equivalent alkalinity in milligrams per liter of the hydroxyl radicle (OH). 1 Fox, C. J. J., On the coefficients of absorption of the atmospheric gases in distilled water and sea- water. Conseil permanent international de la mer, pub. de circonstance 44. February 1909. 7I 72 Papers from the Marine Biological Laboratory at Tortugas. DETERMINATION OF CHLORINE. Chlorine was estimated by means of the salinity outfit supplied by the Copenhagen laboratory of the Conseil permanent international de la mer. This apparatus was obtained through the courtesy of the United States Bureau of Fisheries. The procedure is an adaptation of the usual method of estimating chlorine by titrating with silver nitrate in presence of potas- sium chromate. An essential feature is a sealed tube of standard sea- water whose content of chlorine has been very carefully determined. This water is used for comparison and the apparatus is so constructed and cali- brated as to insure maximum accuracy. Standardized burette No. 8, measuring about 1.5 mm. between graduations, and pipette No. 3, having a capacity of 15.04 c.c., were used. Standard sea-water P; 2/2, 1912, with a chlorine content of 19.386 grams per kilogram, was titrated frequently during the tests at Tortugas. In the later tests comparison was made indirectly with the same standard by means of a large sample of sea-water from Tortugas that had been titrated several times. Knudsen’s correction (k) has been applied to the titrations! and salinity (S) has been computed by means of his formula,! in which Cl represents grams per kilogram of chlorine: S = 0.030 + 1.805 Cl. SPECIFIC GRAVITY. Density was determined by comparison in 10 c.c. weighing flasks with distilled water at 25° C. Knudsen’s values! for pi7.5 were used for com- puting density (S73) as follows S=3 =1-+ pres, where py.5 = 1.00129 (0.1245 + oo — 0.059500 + 0.000155002) and op = — 0.069 + 1.4708 Cl — 0.001570Cl? + 0.0000398 Cl*. Part of the relatively slight difference between determined and computed figures for sea-water around Biscayne Bay may be attributed to difference of standard; that is, by the deter- mined value the sea-water is compared with distilled water at 25° C., while by the calculated value the specific gravity of the sea-water at 17.5° is referred to distilled water of the same temperature. SEA-WATERS AT TORTUGAS. It is believed that solution of calcium carbonate by carbon dioxide in sea-water, if such action takes place, would be shown by regular differences in condition of carbon dioxide in the ebb and flood waters passing out of and into the lagoon surrounded by the shoals and keys of the Tortugas. Accordingly, samples of sea-water were collected twice a day from the middle of Southwest Channel about a mile southeast of Loggerhead Key; these were taken in bottles provided with washered caps that could be clamped down to prevent escape of gases and the samples were kept on ice until they were examined. Those collected from June I1 to 16, inclu- sive, were tested immediately, and though those taken before the former date had been stored for several days the tests do not indicate that the delay in examination had any appreciable effect on their composition. 1 Knudsen, Martin, Hydrographical tables. Copenhagen, 1901. Some Chemical Characteristics of Sea-water at Tortugas. ies TABLE 1.—Salinity and Carbon-Dioxide Content of Sea-Water from Southwest Channel, Tortugas, Florida. o | oh ales Carbon dioxide (CO2). ashe Ss oe 45: AS : Able 2 oi Own} wn a] = a 3 O's oO =5 one 3 be Oo @ jescjesy) ss | 2 g7O | gels ro) ~ VY 0 y/o S bo it) ita) ov ay F aelH 2 ro) = HO=)HOSE 2°90 aed Sea n g S fs SNCS Sell ao 23s 320 Spee 2 a} = FI rt Olleratcy aT £0 3 aug 2 S = xe) S |ek2/sk2| ve 290 5 aqas | 5 QA vu -cccessqcscsaraceancsscsreeness | AF Are (Gol Or arc seacccnecarcnssas satscncesarctsaccerecvocuracsesesesnesseiplccnsenedn ees 32 41 1 Samples collected in June 1913, by R. B. Dole; analyzed by W. T. Read. TESTS OF BAY WATERS. Samples were collected in or just outside Biscayne Bay at the points shown on the accompanying map and the chemical tests of them are re- corded in table 4. TABLE 4.—Some Chemical Characteristics of Sea-Water Around Biscayne Bay, Florida. [Samples collected June 23, 1913, by R. B. Dole; analyses by E. C. Bain.] ' . A -| as . > eh * n= _ -_ 285820) 35 q| 2% | 2% | BeO/Rem) 82) ¢ | ae i ES | Temp. 23-\38>| a3| 8 a B- | Bo No. Location. of Time. es eS 2o3| Sa] 8 3 oy ne water. 3 oas Sap ee 2 a P= fssiaeside|° | 3 | gs] ¥ 2 2 3 S08 SSIS = 3 | ag | as SLE SiO a oO 3) so) Gs Grams per kilogram. I | Mouth Miami River inside bar near Royal Palm dock........ ..ceccccceeeeesees 26.3 4530" a.m. | Tr. |0.232 | 0.078) 1.42) 2.59] 1.0020] 1.0023 II | 1.5 miles east of Miami at 11 ft. depth in government channel north of VAT EIN A PINCV os nage Bg Tames nbeage OE HL ah ak Vials meet taf: am aT eevee STUDIES OF JAMAICA ECHINI. By RoBEert TRACY JACKSON. It was my privilege to join the Carnegie Institution of Washington expedition to Jamaica in the spring of 1912. While there, studies were made of the later stages of development and the variation of a number of species of regular Echini. All the material studied was collected at Montego Bay, close along shore in shallow water of less than a fathom indepth. This locality is very favorable for studying these animals, as abundant material can be obtained, on the reefs or grass-covered bottom, by wading in water usually less than 3 feet in depth. The observations are in extension of those recently published in the ‘ Phylogeny of the Echini’’ (Memoirs Boston Society of Natural History, vol. 7, 1912). As only a few species are in- volved, they are taken up systematically, giving under each the results of the observations. EUCIDARIS TRIBULOIDES (Lamarck). This species is fairly common at Montego Bay, so that sufficient material was available for laboratory work, but it was not so abundant as the other species of regular Echini studied. It occurs in shallow water and is sur- prisingly dull and sluggish, presenting none of the activity of motion seen in Centrechinus or the more passive Tripneustes. In collecting Eucidaris, one simply picks them up, and in no case observed did the animal cling to the ground by its tube-feet. The adults at Montego Bay are rather small for the species, not attaining the size of specimens seen from the Bahamas. An average adult from Montego Bay measures about 30 to 35 mm. in diameter. Material from Port Antonio, Jamaica, also is small, but is very abundant, as I had over 500 specimens collected at that locality. Of a limited number of specimens of this species from the Bahamas, the largest noted is 52 mm. in diameter. Only a few young specimens of Eucidaris were found at Montego Bay. In individuals measuring up to 10 mm. in diameter the ocular plates are all exsert and the genital pores are wanting. There was not sufficient young material to study the periods at which the ocular plates travel in and reach the periproct. In the adult of this species, typically (58 per cent), oculars V, I, IV are insert, as previously ascertained from an examination of 849 specimens from several localities. A specimen measuring II mm. in diameter has all five genital pores in place, as have also all larger specimens. Whether the genital pores would I41 142 Papers from the Marine Biological Laboratory at Tortugas. come in at this exact period of growth could only be known from obser- vations on a larger series of specimens than was available. In living specimens, opened when perfectly fresh, it was found that the teeth extend only very slightly above the base of the foramen magnum. As this struc- ture is very shallow in Eucidaris, the dorsal border of the tooth very nearly coincides with the upper limits of the lantern, but the tooth does not extend horizontally over the top of the lantern as in Strongylocentrotus (Phylogeny of the Echini, p. 179, plate 5, figs. 1,6). The dental capsules, which are the fleshy bags inclosing the base of the teeth, are small in Euci- daris, not large and inflated as they are in Strongylocentrotus and in the Echinide. The Cidaridz, the sole family representing the order Cidaroida, is the only group of modern Echini that extends back into the Paleozoic. The Paleozoic species, which belong to the genus Miocidaris, present only slight differences from recent typical members of the order. As an ancient group that has suffered comparatively little change in its whole geological history the structure and development of the Cidaride is of especial inter- est. Something is known of the post-embryonic development of cidarids, due principally to Loven’s critical study of young Goniocidaris (Echinologica, Stockholm, 1892), but further knowledge of the developing stages of this primitive archaic type is much to be desired. CENTRECHINUS SETOSUS (LESKE). The most abundant sea-urchin at Montego Bay is Centrechinus' setosus (Leske), which abounds in countless profusion in shallow water on the reefs. In strong distinction from the quiescent habits of Eucidaris, Cen- trechinus is a most active animal, moving about freely and actively and waving its long spines in a threatening fashion. The spines are so sharp and cause such poisonous wounds that they are a constant menace to bathers, and specimens have to be collected with caution or serious results will follow. The tube-feet have only the slightest hold on the sea-floor, so that specimens can be picked up with long-handled forceps without any perceptible resistance. In this Centrechinus differs markedly from Trip- neustes, Strongylocentrotus, or Echinometra, which on a rocky surface cling tenaciously by the tube-feet. Young specimens of Centrechinus have spines which are strongly banded; white and very dark, nearly black, alternating. The light and dark bands are of about equal width. The spines are long in young as well as in adults, and a specimen 12 mm. in diameter has spines 36 mm.inlength. A specimen 60 mm. in diameter has spines up to 150 mm. in length. From this it occurs that a small specimen looks rather big with spines extending radially, and a large specimen is menacing indeed when the needle-like sharpness of these weapons is considered. Banded spines as a _ _1Centrechinus is a name which I gave (Phylogeny of the Echini, p. 27) to replace the name Diadema, which in post-Linnzan usage is preoccupied for a crustacean. To replace an old, established name is unfor- tunate, and some objection has been raised to my action. As Diadema has been in current use as a generic name for sea-urchins, Gastropoda, and Lepidoptera, it seems that the rule of priority must be maintained and a new generic name substituted for Diadema in these three groups. Studies of Jamaica Echint. 143 character are retained until individuals are about 20 to 25 mm. in diameter, after which they are typically a deep uniform black. As an exception, a specimen 40 mm. in diameter was seen, in which all the spines were banded asin youth. Frequently in immature specimens the older ventral spines are banded when the dorsal spines, situated on the later-added and therefore younger plates are quite black. In adults, no case of banded spines was seen, but occasionally pure white spines are interspersed with the black, and in a few cases observed all the primary spines were white in the adult. Whatever the color of the spines, the test is of a dark, uniform black in life. In the family of the Centrechinide, Centrostephanus longispinus (Phil- lipi), from the Mediterranean, in adults has spines which are banded white and light brown, the pattern recalling those of young Centrechinus. In Echinothrix calamaris (Leske), from the Pacific, the spines are very variable in adults, being all dark, purple or green, all white, or banded; great diversity may occur in the spines of one individual. In other families of Echini, banded spines occur occasionally as species characters; they are a feature of Celopleurus maculatus A. Agassiz and Clark, and are common in species of the Temnopleuride. In my previous studies of ocular plates and their variation in this species, there was a limited number (110) of developing specimens, and of these very few were really small. Of adults, 1,168 were examined from various localities. With a good series of developing specimens from a single locality, certain differences are brought out from the earlier limited observations. There are also differences in the percentages of adult char- acters from what obtained in the species as a whole from different localities. In table 2 (page 156) are given the characters of developing series, and the variations of adults of this and other species are considered. The aber- rant variations are given in detail only under the consideration of the species in the text. As to the developing series of Centrechinus setosus, in 50 specimens from 4 to 15 mm. in diameter, all the oculars are exsert (fig. 1). In 103 specimens 15 to 20 mm. in diameter, 78 per cent have all oculars exsert, 4 per cent have ocular I insert, and 5 per cent have ocular V insert. In 2 per cent oculars I, V are insert, and in 7 per cent oculars I, V, [V have reached the periproct. Of aberrant variants of this size there are 5 per cent, one specimen having ocular IV only insert; one having I, IV insert; two having V, IV insert; and one has oculars I, V, IV, III insert. It is interesting to see that at this youthful period, while a high percentage have oculars all exsert, many stages of development as regards the oculars are presented by some of the specimens. Of the next larger size, 20 to 25 mm., 270 speci- mens, 51 per cent still have all the oculars exsert; 1 per cent have ocular I insert; and 16 per cent have ocular V insert; 11 per cent have oculars I, V, and 17 per cent have oculars I, V, IV insert. In one specimen, 0.4 per cent, oculars I, V, IV, II are insert. There are eight aberrants, or 3 per cent; of these, 1 has ocular IV insert; 1 has I, IV, and 6 have oculars V, IV insert. 144 Papers from the Marine Biological Laboratory at Tortugas. In the next larger series, 25 to 30 mm. diameter (240 specimens), 21 per cent have all oculars exsert; 3 per cent have ocular I, and 8 per cent have ocular V alone insert ; 14 per cent have oculars I, V, and 45 per cent have oculars, I, V, IV insert. As this last is the species character, it is of note that this is the first stage at which this species feature has become a dominant character. In 1 per cent, oculars I, V, IV, II, and in 1 percent all oculars are insert. Seventeen specimens, or 7 per cent, have an aberrant arrange- ment. Of these, 4 have ocular IV only insert; 11 have oculars V, IV, one has I, V, IJ, and one has oculars I, V, IV, III insert. The occurrence of I, V, IJ insert as an aberrant variant is of interest because this, as previously shown, is a common aberrant when three oculars reach the periproct in the Echinide and Strongylocentrotidz, and occurs also in the Stomopneustide. I have not found a case of I, V, II insert in the Cidaride, and it is the only case seen in Centrechinus setosus, although in this species, 1,398 cases of I, V, IV insert have been observed. In the next series of Centrechinus setosus, 30 to 35 mm. diameter, 245 specimens, Io per cent have all oculars exsert; 2 per cent have I only; and I2 per cent have V only insert; 5 per cent have I, V, and 63 per cent have I, V, IV insert; 4 per cent have I, V, IV, II insert. Of aberrants, there are 4 per cent, these consisting of 8 specimens with oculars V, IV insert, and 2 with oculars I, V, IV, III insert. The next series, measuring 35 to 40 mm. in diameter, 169 specimens, has 5 per cent with all oculars exsert; 2 per cent with I only; and 3 per cent with V only insert; 5 per cent have I, V, and 71 per cent have I, V, IV insert; 6 per cent have I, V, IV, II, and 2 per cent have all ocularsinsert. Aberrants are 5 per cent; of these, two specimens have ocular IV alone insert; 1 has I, IV; 4 have V, IV; and 2 have I, V, IV, III insert. The series 40 to 50 mm. in diameter is considered the last of the devel- oping series. Of this size, 162 specimens, 4 per cent have all oculars exsert, I per cent have ocular I, and 0.6 per cent have ocular V only insert; 6 per cent have oculars I, V, and 70 per cent I, V, IV insert; 9 per cent have oculars I, V, IV, II, and 6 per cent have all oculars insert. Of aberrants there are 3 per cent, one specimen having V, IV and 4 having I, V, IV, III insert. The series 35 to 40 mm.and 40 to 50 mm. in diameter make a close approach to the mature series as regards the species character of I, V, IV insert, but these two younger series have more of the developing and fewer of the progressive characters than are seen in the older series. While Centrechinus setosus is profusely abundant at Montego Bay, specimens do not attain as large a size as in Florida, from which locality many individuals exceed 100 mm. in size. Of the adult series from Montego Bay, 50 to 80 mm. in diameter, 162 specimens, one specimen, or 0.6 per cent, had ocular V only, and one had oculars I, V insert, both of these specimens are between 50 and 60 mm. in diameter; 73 per cent have I, V, IV insert, the species character; 13 per cent are progressive variants with I, V, IV, II insert and 10 per cent are extreme progressive variants with all oculars Studies of Jamaica Echint. 145 insert; 3 per cent are aberrant variants, all five specimens having oculars I, V, IV, III insert. Considering the ocular-plate arrangement in Centrechinus from Montego Bay as a whole, it is seen, as shown clearly in table 2 (page 156), that from all exsert,the character of the young, there is a steady progressive series with growth in the development or traveling in of ocular plates. Centre- chinus is a member of the most primitive suborder, the Aulodonta, of the Centrechinoida, and as such is worthy of special attention, for, as shown by Professor Hyatt, primitive types in a group have a slow development as com- pared with specialized types, and in character approach nearest to the next lower series of their own phylum. The development of Centrechinus amply bears out this important truth. Of the total 1,401 specimens of Centrechinus setosus listed from Mon- tego Bay, the aberrants are 59 in number, of which 8 have ocular IV only insert; 3 have oculars I, IV insert; 32 have oculars V, IV insert; 1 has oculars I, V, Il insert, and 15 have oculars I, V, IV, III insert. It is note- worthy that in the mature series (50 to 80 mm. diameter) the only aber- rants that occurred were cases of oculars I, V, IV, III insert, thus indicating that the other aberrants seen in immature individuals may be considered as irregularities in the order of sequence of the coming in of ocular plates, rather than as aberrants that would have retained the given character if they had lived to grow up. In specimens of Centrechinus up to 14 mm. in diameter (48 individuals), all the genital plates are imperforate (fig. 1), whereas in those 14.5 mm. and all larger with very few exceptions, all genital plates have the genital pores.’ In comparison with this, the specialized Strongylocentrotus drébachiensts attains its genital pores when between 5 and 10 mm. in diameter (Phylogeny of the Echini, pp. 131, 170). Centrechinus has all oculars exsert up to 15 mm. diameter, and with intermediate stages still has 4’ per cent all exsert when 40 to 50 mm. diameter. Sfrongylocentrotus drébachiensis, on the contrary, has typically oculars all exsert only in very young specimens and has only 29 per cent with all exsert in specimens (51) between 2.5 and 4 mm. in diameter; further, it has only 4 per cent with oculars all exsert in specimens (82) between 4 and 5 mm. diameter. Centrechinus first attains the species character of oculars I, V, IV insert as a dominant feature (45 per cent) when it is 25 to 30 mm. in diameter. Strongylo- 1 centrotus drébachiensis attains I, V insert, the ™~ ine Corpreoanes one Cee species character, as a dominant feature (52 per oer gg | oege cent) when 5 to 10 mm. in diameter. In Centre- ee eit cnooeae 1] regret to say that by error in the figure published of a young Centrechinus (Phylogeny of the Echini, fig. 88, p. 106), genital pores are shown. The specimen has no pores and they were inadvertently drawn. 10 146 Papers from the Marine Biological Laboratory at Tortugas. chinus when oculars begin to enter the periproct, the first to enter is V or I, but in the Montego Bay series V is by far the more common,’ they being in the ratio of 102 of ocular V to 23 of ocularI. In this feature Centrechinus makes an approach to the Cidaride, in which group, when one ocular is insert, it is almost always ocular V. As previously shown, when in the Centrechinoida four oculars reach the periproct, it is typically I, V, IV, II, and when any other combination exists it is nearly always I, V, IV, III. This combination, while rare in the Centrechinoida as a whole, is rela- tively more frequent in Centrechinus and is a typical feature or a frequent variant in some species of the Cidaride. In this feature also, therefore, Centrechinus makes a certain approach to the Cidaroida. In Centrechinus, when opened fresh and alive, it is found that the teeth extend above the base of the foramen magnum about to the upper line of the lantern, but the proximal base of the teeth does not extend horizontally over the lantern as in the Echinide, Strongylocentrotide, and Echinome- tride. Further, the dental capsule, inclosing the base of the tooth, is small. In the limits of the teeth dorsally and the small dental capsule, as well as in the grooved teeth, Centrechinus makes a close approach to the characters of the Cidaride as represented by Eucidaris tribuloides. Fics. 2-6.—-Perignathic girdle in Aspidodiadema and its development in Centrechinus. 2.—Adult Aspidodiadema meijerei (Déderlein), Pailolo channel, Hawaiian Islands. The auricles are separate spur-like styles. 3.—Cenirechinus setosus (Leske), Montego Bay, Jamaica. Specimen 5 mm. diameter. Auricles are separate styles, apophyses faint. 4.—The same. Specimen 10 mm. diameter. Auricles are more developed and arch over the ambulacra. 5.—The same. Specimen 25 mm. diameter. Auricles meet over the ambulacra in a slender arch. Sara save ees 70mm. diameter. Auricles developing a high plate-like character, apophyses also are high. 7.—Centrechinus setosus, Bermuda. Specimen 110 mm. diameter. Auricles have developed a high plate-like character; apophyses are high ridges. Very little attention has been paid to the development of the perignathic girdle excepting by Lovén, who described it in Cidaris and Strongylocentrotus (Echinologica). I also have published a few observations on the develop- ment and variation of the perignathic girdle. Centrechinus, while being a primitive type in many respects, has in the adult a specialized perignathic girdle having high auricles which meet in a broad plate-like arch over the ambulacra and also high apophyses on the interambulacral areas (fig. 7). Such being the character of the adult, it is of much interest to follow the development of this, which is one of the most specialized perignathic girdles known in Echini. When a specimen is 5 mm. in diameter, the apophyses show only slight indications of development and the auricles stand up as separate, blunt, spur-like processes on either side of the ambulacrum (fig. 3). 1 In previous observations from various localities with limited material, ocular I predominated over V numerically. Studies of Jamaica Echint. 147 The upper ends of the auricles at this early stage are slightly flattened, but do not arch over the ambulacrum at all. Inaspecimen 10 mm. in diameter, the apophyses have developed somewhat, and the auricles are inclined over the ambulacral area, but do not meet (fig. 4). In specimens 25 mm. in diameter the apophyses have developed into distinct elevated ridges, and the auricles have met over the ambulacral areas as slender arches (fig. 5). From this point on to the adult the development of the perignathic girdle is marked by the increase in height of the apophyses and concurrently the increase in height and the lamellar expansion of the auricles (figs. 6, 7). In seeking comparisons to these stages of Centrechinus in the adults of related forms, it is found in Aspidodiadema metjeret (Déderlein), A. nico- baricum Déderlein, and Dermatodiadema horridum A. Agassiz, representing the family Aspidodiadematidz, that the auricles exist as small conical spurs situated on either side of the ambulacral areas, and apophyses are not developed (fig. 2). This condition corresponds with the first stage noted in the development of Centrechinus (fig. 3). This relation of the perignathic girdle is of much interest because in the Aspidodiadematide the simple ambulacral plates, the large primordial ambulacral plates on the peristome, the large apical disk and simple spur-like auricles are all primitive characters, indicating that the family is more primitive structurally than is the still primitive family of the Centrechinide. While primitive in many char- acters, the Aspidodiadematidz is specialized in that all the oculars reach the periproct, and Centrechinus, while belonging to the primitive suborder Aulodonta, is specialized in its great development of the perignathic girdle. This shows how primitive and specialized features may be combined in one and the same type. Mr. Agassiz (Panamic Echini, 1904, p. 59) says of the Aspidodiade- matide: ‘‘The auricles are most irregularly developed. They are either wanting or mere projections, slightly raised.’’ I have examined three rep- resentative species and a number of specimens of this family, and find no evidence that the auricles are irregularly developed. In adults they are not wanting except where broken off. They are perfectly definite spur- like projections, and of much interest as being structurally the most primi- tive auricles known in any of the adult Centrechinoida. As the Cidaroida have apophyses but no auricles, and as these structures make their first known appearance in the Centrechinoida, the simplicity of their structure in adults of the Aspidodiadematidz and in the young of Centrechinus is of particular interest. TOXOPNEUSTES VARIEGATUS (Lamarck). A common species at Montego Bay is Toxopneustes variegatus (Lamarck), which is found abundantly on the grass-covered bottom in shallow water. Specimens attain a large size, some measuring 70 mm. in diameter. Few young specimens were found, though this is perhaps due to the fact that special search was not attempted, from limitations of time. The test is 148 Papers from the Marine Biological Laboratory at Tortugas. mottled green and white, with green spines as the prevailing color. Some young specimens show on the green and white groundwork purple pri- mary tubercles and purplish spines. A peculiar color-pattern was observed as a relatively rare variation in the fact that in the interambulacral areas of some specimens the green followed a zigzag pattern on a white ground and extended thus from the apical disk adorally, but in no case observed extended further than the ambitus. The zigzag green color- pattern on a cream-white ground of a specimen measuring 45 mm. in diam- eter is shown in fig. 8. The lower figure on the same plate is of a somewhat larger specimen, measuring 48 mm. in diameter, in which the zigzag pattern occurs only near the apical portion of the test. Both of these specimens with others figured in this paper are now in the collections of the Museum of Comparative Zoélogy, at Cambridge, Mass. This type of color-pattern was quite new to Dr. Clark and Professor Tennent, both of whom had worked on the species, and I had never seen it before, although I have studied many specimens from different localities. Being busy with other species, no attempt was made to collect a large series of Toxopneustes variegatus for a study of ocular plates, but 400 speci- mens are tabulated. These are of various sizes, but none were young and they were not graded into sizes for comparison. Of this series, 2 specimens, 0.5 per cent, have ocular I only insert as arrested variants, and one specimen, 0.3 per cent, has ocular V only insert; 84 per cent have oculars I, V insert (figs. 8, 9), which is the species character in all localities; 15 per cent are pro- gressive variants with oculars I, V, 1V insert. Four specimens, I per cent, are aberrant variants and all of these have oculars I, V, II insert. The ocular character is practically the same as I showed previously in this species from several localities, but the Montego Bay material has a somewhat lower percentage of I, V and corresponding higher percentage of I, V, IV insert. My previous observations on 1,043 specimens gave go per cent I, V insert and 8 per cent I, V,IV. Comparison of the tabulation of this species may be made with that of the closely allied Toxopneustes atlanticus (A. Agassiz) from Bermuda, in which I showed (Phylogeny of the Echini, p. 161) that the mature series (45-77 mm. diameter) has a much higher percentage (28 to 29 per cent) of oculars I, V, IV insert than has Toxopneustes variegatus. The Montego Bay experience in the cases of several species brought out the fact clearly that for a close study of variation it is very desirable to tabulate ocular plate characters in specimens from a single definite locality because considerable difference in a given species may occur in different localities. In Toxopneustes variegatus, when opened fresh, it is found that the proxi- mal part of the teeth extend horizontally over the top of the lantern and the teeth are bent back on themselves in the same plane, also the dental capsules which inclose the base of the teeth are large and bladder-like, in these char- acters differing from Eucidaris and Centrechinus, but agreeing with Strongy- locentrotus as I previously described it. In a young specimen 12 mm. in diameter, the auricles are still separate, but in larger specimens they are JACKSON Studies of Jamaica Echint. 149 joined in anarch over theambulacral areas. In large adults the apophyses are developed as a moderate ridge and auricles are joined in a delicate arch which never becomes heavy, as in large specimens of Tripneustes. TRIPNEUSTES ESCULENTUS (Leske). One of the abundant species at Montego Bay is Tripneustes esculentus (Leske). It occurs on grass bottom and also on the reefs in shoal water, and is a strikingly handsome sea-urchin when alive, with its white spines against the dark purplish groundwork of the fleshy covering of the test. The larger individuals live out in the open in full view, but young specimens are found only in crevices or on the under side of rocks. The young are not common, and although sought for diligently, no very young specimens were found and only a limited number of the youngest seen were obtained. Perhaps some other time of year, as summer or autumn, would be more favorable for young material of thisspecies. In Tripneustes esculentus from this locality the ocular plates continued to travel in or enter the periproct, from the youngest to the largest series of specimens observed, as shown by the tabulation. For this reason, as regards the feature of ocular plate development, what is called the developing series for this species in the table is arbitrarily drawn at a maximum of 70 mm. Taking up the developing series: Ten specimens of 20 to 25 mm. diam- eter have 50 per cent with ocular I only insert, and 50 per cent with oculars I, Vinsert. Younger specimens would doubtless have all oculars exsert as a developing stage. The next size, 25 to 30 mm. diameter, 21 specimens, has 24 per cent with ocular I only insert and 76 per cent with I, V insert. The relations of these two younger series shows that it is a period of rapid development. The next series, 30 to 40 mm. diameter, 60 specimens, has 12 per cent with I only insert; 80 per cent with I, V insert; 2 per cent with I, V, IV and 2 per cent with I, V, IV, II] insert. Three specimens, 5 per cent, are aberrant with I, V, Il insert. This stage, while holding the I only insert as a developing character, has attained the I, V; the I, V, IV; and the I, V, IV, II insert; which are each respectively the feature of the species as a character in one or more of the several localities from which material was studied, but the relative proportions of the three characters differ from that of any known adult series. The series 40 to 50 mm. diameter, 42 specimens, has lost the I insert as a youthful character; 90 per cent have oculars I, V insert; 2 per cent have I, V, IV, and 2 per cent have I, V, IV, II insert. Five per cent are aberrant, one of the two specimens having I, V, II] insert and the other V, II insert. This stage has the highest percentage of I, V insert of any age of material from Montego Bay. The series 50 to 70 mm. diameter, 34 specimens, has 53 per cent with oculars I, V insert; 18 per cent with I, V, IV; and 6 per cent with I, V, IV, II insert. Aberrants are 24 per cent, eight specimens, all of which have oculars I, V, II insert. The developing series in its later phases may be compared broadly with material from Bermuda or other localities 150 Papers from the Marine Biological Laboratory at Tortugas. for the species in which oculars I, V insert is the adult local character, as it is the youthful character in these immature stages from Montego Bay. The next larger series, 70 to 90 mm. diameter, 125 specimens, has one specimen with ocular I insert as an arrested variant; 30 per cent have oculars I, Vinsert. The latter, which was the leading character in the later developing stages, has now passed into the phase of an arrested variant. 34 per cent have oculars I, V, IV insert, and this is the dominant character of this age. As a phase it can be compared with a series of adults from the '®, Aa . . SAN ai lif — oy + ow, _@ - oF 13 14 Fics. 10-14.—Normal ocular plate arrangement in Tripneustes esculentus (Leske), Montego Bay, Jamaica. All figures nearly X 2. 10.—Diameter 99 mm. Ocular I only insert, an extreme arrested variant. I1.—Diameter 114 mm. Oculars I, V insert, an arrested variant for the locality. 12.—Diameter 110mm. Oculars I, V, IV insert, an arrested variant for the locality. 13.—Diameter 115 mm. Oculars I, V, IV, II insert, the typical character for the locality. 14.—Diameter 106 mm. All oculars insert, a progressive variant. Bahamas in which locality I, V, IV insert is the typical character. Twenty- eight per cent have oculars I, V, IV, II insert, which at this stage may be considered a progressive character as it is at all other known localities for the species. In 8 per cent the ocular arrangement is aberrant, all of the ten specimens having oculars I, V, II insert. The next series, 90 to 110 mm. diameter, 200 specimens, has one speci- men with ocular I only insert as an arrested variant (fig. 10), and one specimen with ocular V alone insert. For ocular V only to be insert is a rare variation in Tvipneusies and is unusual in the family Echinide. In 22 per cent oculars J, V are insert. This, which was the character of the later developing series and is the character of adults at Bermuda and Florida, has dropped considerably in its percentage as an arrested variant. In 24 per cent oculars I, V, IV are insert. This character for Montego Bay has Studies of Jamaica Echini. I51 at this stage dropped into the phase of an arrested variant, though it was the dominant character in the series 70 to 90 mm. in diameter and is the adult character in a series from the Bahamas. Forty per cent have oculars I, V, IV, Il insert. This is the first series in which this character is a domi- nant feature at Montego Bay, and for all earlier stages in this locality and for all other localities known it is a progressive character. Two per cent have all oculars insert as a progressive variant (fig. 14). It is somewhat striking that with four oculars insert as a dominant character, there should not be more specimens with all oculars insert. In this series, 25 specimens, 13 per cent, have an aberrant arrangement of ocular plates. Of these aberrants, 22 have oculars I, V, II insert; two have I, V, IV, III insert (fig. 16), a very rare variant in the Echinide when four plates reach the peri- proct, but although rare here, it is a common character in the Cidaridz and a not rare variant in Centrechinus setosus. One aberrant specimen of Cs ney SS aay N yy 15, 16 FIGs. 15-17.—Aberrant ocular plate arrangement in Tripneusles esculentus (Leske), Montego Bay, Jamaica. All figures nearly X 2. Fic. 15.—Diameter 115 mm. Oculars I, IV, II insert, a common aberrant variant, a right-handed equiva- lent of the normal character I, V, IV insert. Fic. 16.—Diameter 106 mm. Oculars I, V; Ws III insert, a rare aberrant variant, but the usual one when four oculars but not I, V, IV, II are inse Fic. 17.—Diameter 102 mm. Oculars LoVe IL “III insert, a unique aberrant variant, a right-handed equiva- lent of I, V, IV, III insert. Tripneustes has oculars I, V, II, III insert (fig. 17). This, which is a right-handed equivalent of I, V, IV, III Gust as I, V, II is a right-handed equivalent of I, V, IV) is a unique variant for Echini as far as my experi- ence goes. In the Centrechinoida, as tabulated in my preceding memoir and in the present paper, 619 specimens are recorded, in which four plates reach the periproct. Of these, in 554 specimens (89.5 per cent) it is oculars I, V, IV, II the bivium and posterior pair of the trivium, in 64 specimens (10.3 per cent) it is the aberrant arrangement I, V, IV, III, and in the one specimen cited (0.16 per cent) it isI, V, II, III. (Compare figs. 13, 16, 17.) The series containing the largest specimens, I10 to 132 mm. diameter, 82 specimens, has 13 per cent with oculars I, V insert as arrested variants (fig. I1); 24 per cent with I, V, IV insert as arrested variants (fig. 12); and 43 per cent with I, V, IV, II insert as the local character of the species (fig. 13). Five per cent have all oculars insert as a progressive variation, and 15 per cent, I2 specimens, are aberrant in ocular arrangement. Of these 152 Papers from the Marine Biological Laboratory at Toriugas. aberrants, II specimens have oculars I, V, II insert (fig. 15), and one has V, IV, Il insert. In this specimen genitals 5 and I are fused, mechanically shutting out ocular I from access to the periproct. The series of Tvipneustes esculentus from Montego Bay is very interest- ing from the point of view of ocular development on several accounts. The specimens show a direct progressive increase in the number of oculars insert and the relative percentages of the same from the youngest to the largest series observed. In specimens fully matured, as regards this character, and including from 90 to 132 mm. in diameter, they have as the dominant character (40 to 43 per cent) the ocularsI, V, IV, II insert. This is the only locality known for this species, and it is the only recent species known! in which four oculars reach the periproct as a typical character. In pre- vious studies of Tripneustes esculentus, many specimens were tabulated from several localities, but without detailed measurements of the size of the specimens. As the material was from various sources, it is impossible to go over it again to ascertain the sizes but it is fair to say that the material was practically all adults. With this qualification it is interesting to com- pare the tabulations of Tripneustes esculentus from the several localities from which there was sufficient material to give any reasonable basis for con- sideration. (LABLE wie = Oculart | Ocular y | Oculars 1, | Oculars 1, | Oculars 1, | Arrange- 8 insert; Vv, insert; 1, Vv insert; |V,IvVinsert;| Vv, IV, II All oculars | ment of < I IV, Il, III IV, I, II IV, II, III II, I ex- | insert;mr | insert. | oculars es Locality exsert. exsert. exsert. sert. exsert. | aberrant. vo = si | Qa | | | ag p. ct. No. | p. ct. No. | p. ct. No. | p. ct.| No. | p. ct.| No. | p. ct.| No. |p. ct.| No. = Fal | a re = 2 | =I ia | | | 193 | Bermuda..| 2 | EY uilnoped | 56.008 6r || Ir7 35 G77 | a2 3 |. 05 I I 2 Tan ehlonidase sl eeeaic eae. x I 46 SZ) 201) 6335) 12 re oleate Bete e | 12 13 160 | Bahamas..|..... heer 0.6 I 22 35 | 46 | 73 | 27 420 Phe i) oe 3 5 282 | Jamaica! 0.4 | I 0.4 I 19 54 24 67 | 40 | 114 3 | 8 |} 13 37 | 1 190 to 123 mm. diameter. As shown in table 1, Tripneustes from Bermuda is characterized strongly, 61 per cent, by having oculars I, V insert, with 35 per cent I, V, IV insert as progressive variants, also in 2 per cent I, V, IV, II, and 0.5 per cent all oculars one insert as progressive variants. The aberrants are few, I per cent, one specimen having I, V, II and one having V, IV, II insert. This Bermuda lot is the most primitive of the species that I know of as regards ocular arrangement in adults. Tripneustes from Florida has 46 per cent with oculars I, V insert, which is still the dominant character, but with a lower percentage than at Bermuda. As progressive variants, 29 per cent have I, V, IV insert, and 12 per cent have I, V, IV, II. The aberrants are 12 per cent, all being cases of I, V, II insert. Florida specimens, while having oculars I, V insert as the character, have a lower percentage of this character and a higher percentage of progres- _ lIn Acrosalenia pseudodecorata Cotteau, from the Bathonien of France, oculars I, V, IV, II are said to be insert as a species character (Phylogeny of the Echini, p. 112). Studies of Jamaica Echini. 153 sive variants than Bermuda material, hence it is structurally an advance on that and a step towards the next higher advance as represented in the series from the Bahamas. Material from the Bahamas collected by C. J. Maynard has only 22 per cent of I,Vinsert. This is not the character of the locality and has therefore passed into the phase of an arrested variant. There are 46 per cent with ocularsI,V,IVinsert. This is the dominant character of the Bahama mate- rial, while it would be a progressive variant for Bermuda and Florida, and further, would be an arrested variant for the Montego Bay series. In the Bahamas, 27 per cent have I, V, IV, II insert, a common progressive variant, and 2 per cent have all oculars insert. The aberrants are relatively few, 3 per cent, four of the aberrants having I, V, II insert and one having I, V, IV, III insert. Structurally, in its percentages of ocular arrangement, the Bahama material is intermediate between the Florida and Montego Bay series. The Montego Bay material, considering only the two largest-sized series tabulated on page 156 and treating them as a single series of 90 to 132 mm. diameter, has 19 per cent with oculars I, V insert as a moderately common arrested variant, but with the lowest percentage of any of the localities given. In 24 percent oculars I, V, IV are insert, which, for this locality, is an arrested variant, though it is a progressive variant for Bermuda and Florida, and a dominant character for the Bahamas. In 4o per cent, oculars I, V, IV, II are insert, the dominant character for this locality, though it is a progressive variant for all the other localities; 3 per cent have all the oculars insert, which as a progressive variant, is only a slight advance on that existing in the other regions cited. The aberrants, 13 per cent, have already been considered above, and do not need further mention excepting that it is interesting that most of them are cases of I, V, II insert, as in other localities. In all the localities given Tripneustes esculenius is an abundant form, and in all attains a large size. The specimens from Bermuda were collected for me and averaged very large, the largest measuring 145 mm. in diameter, yet this locality has the lowest ocular index. The cause of the progressive evolutionary series as marked by the higher and higher percentages of ocular plates reaching the periproct is unknown, but it is a fact that it exists and points to the desirability of studying large series of specimens from different localities. In Tripneustes esculentus, when opened alive, it is seen that the teeth extend above the top of the lantern and horizontally over the top of the same. The proximal end of the tooth is curved back on itself and is grooved, the keeled character developing as one passes orally along the tooth. The dental capsules are relatively large, all this structure coinciding with that previously described in Strongylocentrotus drébachiensis. In a young specimen of Tripneustes, 24 mm. in diameter, the auricles are slender, but already meet in an arch over the ambulacra. In large adults the auricles 154 Papers from the Marine Biological Laboratory at Tortugas. are quite massive and are fused over the ambulacra in a long median symphysis. They may attain a height of 14mm. or more. The apophyses are also relatively high and massive in large specimens. ECHINOMETRA LUCUNTER (Linné). Close in to shore on the coral rock at Montego Bay and in very shallow water Echinometra lucunter (Linné) occurs in abundance. Not only adults but individuals of all sizes, down to very young, live in the same exposed situation, where they get the full beat of the waves on shore. This species clings tenaciously to the rock and has to be pulled off from the rock with some effort. The ocular development of this species is very interesting, but unfortunately this was not appreciated until it was too late to gather more material and only a limited series was collected. These were supple- mented by a few specimens collected at the same time by Mr. G. M. Gray. A large series could easily be obtained in this locality. As Echinometra lucunter is more or less elliptical in horizontal outline, being elongated in the plane of the axis of interambulacrum 3 and ambulacrum I, the measure- ments of sizes are given in length instead of in diameter, as in the other species considered. Considering the developing series of Echinometra, a set of 34 specimens measuring 4.5 to 15 mm. in length have all the oculars exsert. Of this series, those from 4.5 to 10 mm. in length (26 specimens) were all wanting in genital pores, but these pores exist in all the larger sizes examined. The series 15 to 20 mm. in length, 23 specimens, has 91 per cent with all oculars exsert and 9 per cent with ocular V insert. While ocular I is the first to enter the periproct in the Echinide and Strongylocentrotide, it is ocular V that typically enters the periproct first in the Arbaciidz and the Echino- metridz, thus indicating family distinctions in ocular development. In the series 20 to 25 mm. in length, 58 specimens, in 71 per cent oculars are all exsert and 29 per cent have ocular V insert. In the 25 to 30 mm. series, I10 specimens, 34 per cent have oculars all exsert; 2 per cent have ocular I insert. These are the only cases with ocular I only insert found in Montego Bay material and it is a rare variation in the genus and family, as previously shown. In 54 percent ocular Visinsert. This series is the first stage in growth in which the typical species character of V insert is a dom- inant feature. In 10 per cent, oculars V, I are insert and in the limited number of specimens this is the first stage at which this character appeared. With more material some specimens would probably be found having I, V insert at an earlier stage of growth. ‘This series is considered the last of the developing stages, but the line as regards ocular development is arbitrary because, as seen in the later series, oculars continue to come in with increas- ing size of the individuals. Of the series 30 to 40 mm. in length, 175 specimens, 17 per cent have oculars all exsert as arrested variants, 58 per cent have ocular V alone insert as the typical species character, and 25 per cent are progressive Studies of Jamaica Echint. 155 variants with oculars V, I insert. The percentage of V insert at this age is quite near that of the next larger series, but there are more arrested and fewer progressive variants than in the older series. The series 40 to 52 mm. in length, 103 specimens, includes the largest specimens collected at Montego Bay. From other localities specimens may attain a length of 90 or more millimeters. Of this series, 7 per cent have all oculars exsert as arrested variants; 66 per cent have ocular V insert as the maximum development of the species character in the locality; 26 per cent have oculars V, I insert as progressive variants. One specimen, I per cent, has oculars V, II insert as an aberrant variant, and this was the only aberrant found in the whole series of 503 specimens collected at Montego Bay. Considering the ocular arrangement of Echinometra lucunter at Montego Bay asa whole: It is a desirable type to study because its species character is to have one ocular insert, and this is a very unusual feature in Echini, otherwise occurring only in the recent Saleniide and in Echinus magel- lanicus Philippi from South America. Other Echini, both recent and fossil, are characterized as the species feature of adults by having all oculars exsert, or two oculars insert, or three or more insert, usually three or five. Five insert is a not unusual feature in both living and fossil types as far back as the Paleozoic. In Echinometra lucunter it is seen that oculars are all exsert up to a comparatively large size, 15 mm., after which oculars travel in gradually with increasing size, the highest percentages of insertness being in full-grown specimens. Comparing the present tabulation of this species with that previously published (Phylogeny of the Echini, p. 163), it is seen in both that ocular V insert is the species character, though in Bermuda material oculars V, I insert is nearly as frequent. In both lots ocular I insert is a rare feature, and aberrant ocular arrangement is rare. As material previously tabulated was not graded by size, no comparison can be made in this respect. j 20 21 Fics. 18-21.—Development of the perignathic girdle in Echinomeira lucunter (Linné). FIG. poe Vouns specimen from Montego Bay, Jamaica. Specimen 5 mm.long. Auricles are erect separate styles. FIG. soa same. Specimen 8 mm.long. Auricles arched over the ambulacrum but separate, apophyses eveloping. Fic. 20.—The same. Specimen 15 mm. long. Auricles joined in suture over the ambulacrum. | : Fic. 21.—Large adult from Bermuda. Specimen65 mm.long. Auricles produced vertically as high spoon-like plates, apophyses high ridges. The teeth of Echinometra lucunter extend horizontally over the top of the lantern and the proximal end of the tooth is bent back on itself and in the same plane, as previously described in Strongylocentrotus. The proximal part of the tooth is grooved, the keel developing as one passes adorally; Papers from the Marine Biological Laboratory at Tortugas. 156 zs ov ov of of Sz Sz oz 0% SI $103 S°7 “yysUeT i L eee eele eee el eee eel eo eee e te eee leer eei see eesisceeee or OL eeeeelecees Ss tz eeeeeleeeee ofc Sz Iz eeeert scorer oes eei eee esi steerer /eeeer eevee ei (sevens Ss os eeeeel eeeee s os eeeereleeeee Sz oz ol = a aeaaa||Poson ooac jones wee) gg | s1| see | tela |@o0 |e |So [el s = ob or Iz £1 | gir fz I 970 I (Oe ea EO vs ee eee 9I eeeee ov ener 6rs sees L6 eevee zZol eee €z s £ or 9 SI 6 11 oL or 9 I 9°0 Z 6 Si v z II 9 ozI IL 6 s s £ £ z or v OOD PO IIE| aCe) Ban BSrs) So) er s 6z zr} V z LI L z I £ I 601 St Tee vr | 61 8 9 £ 1s Iz Of ase ova 8 £ ar eas ee ae) ov LI | of Ir | eV oI] v I ger | 18 Se 02 oLz S$ S$ sere rlscooes{seeerieeene L Z Zz t Ss ¢ | v v og gl oz cI for Oe eo ee ee eee ee ee ee ee ee er a er eer ee ery eoeelecece SI 01? os ‘ON |"39°d| con |'90°d | ‘on |*99°d| con |g0-a | sony [39a | “ony [99-4 | con |*39°d| ‘ont [90-4 *pearos “yUeLIeqe *y1osxo “yesxo *q198x9 *ql9sxo *qyosxo ‘ d -qo sre[noo *}osul III $4.19sut Ill ‘II III ‘II ‘AI Ill ‘II ‘AT Ill ‘II ‘AT *yIosxo ho eg eet) suoul jo Juour sie[nso [Ty Tl ‘Al ‘A !yesurAI‘A| [929SUT A “I qesul ‘A SyZesur | sre[nso [TV “Deds ~osueliy ‘I SIe[NIO | ‘I sxejndQ | ‘I siejndIO A re[ndOD I 1ejnd09 } + tse++sesgoras padopaad glz I *s9+** ssarras BuIdojaAoq Sez S49JUNIN] DAJQULOULYITT vLs } tr es++*sgarzas padopaaq Lov } tees +sorag SurdopeAed LOI isnquajnasa $ajsnaugt4 J, ‘8 ***-onipsataDa Sajsnaugoxoy, | ooVv 10v'T SOD ODOC GR Var: pedopasq zor theese ssoriag SuIdojaaaq Of2‘I isnsojas snu1yIadjuad *“suoul *solias pue soods =e T : jo ‘ON Te}OL ‘papuor ‘kong odajuopy wmosf wiyry fo uoyvrapa pup juamasunsdp aypjd 4njnI0 yo91dk[—z AAV], Studies of Jamaica Echini. 157 the dental capsules are large, embracing the base of the tooth as in the Echinide and Strongylocentrotide. The perignathic girdle of Echino- metra lucunter in the adult is highly specialized, having relatively high ridge- like apophyses and extravagantly developed auricles which meet over the ambulacra and are produced dorsally as spoon-shaped plates with a long median symphysis (fig. 21). In young specimens, 5 mm. in length, the auricles exist as simple erect styles situated on either side of the ambulacra but not arched over the same (fig. 18). In specimens 10 mm. in length, the auricles arch over the ambulacra, but are still separate, not meeting in median suture (fig. 19). Ina specimen 15 mm. in length, the auricles have met over the ambulacra and are joined by median suture, and the apophyses exist as moderately developed ridges (fig. 20). From here on to the adult condition the apophyses increase in height and the auricles gradually take on an extensive vertical enlargement of their upper border (fig: 25) 2m Echinometra viridis A. Agassiz, E. oblonga Blainville, and E. vanbrunti A. Agassiz, in adults, the auricles are relatively slender, but are united in suture over the ambulacra; there is no vertical extension in a spoon-like fashion, however, and the auricles closely resemble the condition of E. lucunter when about 15 mm. in length. In Echinometra mathei Blainville, in a young specimen 8 mm. long from Mauritius, the auricles are still separate, but in the adult the auricles are joined in suture and with moderately developed spoon-like processes, but not as high processes as are those in adult E. Jucunter. In Heterocentrotus érigonarius (Lamarck), from Mauritius, a young individual 9 mm. in length, has the auricles still separate, but in the adult of this species auricles are arched and united over the ambulacra and with moderately developed spoon-like processes. SUMMARY AND CONCLUSIONS. Montego Bay, Jamaica, is an excellent locality for studying Echini on account of the number of species there available and the abundance of material. It is an ideal place for a student who wishes to supplement studies on northern species by observations on tropical forms. Some 2,878 specimens of regular Echini were collected and studied, but in no case did any specimen show a departure from the typical pentamerous system. This is mentioned as in previous studies it was found that, on the average, completely or partially trimerous, tetramerous, or hexamerous ‘specimens occurred a little oftener than once in a thousand specimens. In the ‘ Phylogeny of the Echini,”’ page 91, it is stated that ‘‘all the evi- dence goes to show that the full number of oculars that are to become insert are developed early in the life of the individual and apparently later no change in this respect takes place.” It was assumed that in Echini specimens of about half the mature size could be accepted as showing the mature characters as regards oculars, and the tables of ocular plate vari- ation, while based principally on adults, were made up on the basis of con- 158 Papers from the Marine Biological Laboratory at Tortugas. sidering specimens more than half-grown as being fully developed in this respect. This view was based largely on studies of Strongylocentrotus dré- bachiensis, and in this species the view was fully justified. It is doubtless true of many species, perhaps most species of Echini, that specimens half- grown have practically the full character as regards ocular development. It is not entirely true, however, of all species, for as shown in this paper, in Centrechinus setosus, Tripneustes esculentus, and Echinometra lucunter (table 2), ocular plates continue to come in or enter the periproct as a developing character until nearly or quite grown; therefore, as a result in these species, the largest numbers in percentage of oculars insert may be found in the series of largest specimens. This is not true of selected indi- viduals. That is, the largest specimen does not necessarily have more oculars insert than a smaller individual, but taking a large series of indi- viduals of these species, as shown in the table, the average of each size up to the largest has more oculars insert than any of the preceding sizes. This late coming in of oculars in some species is of interest as showing the mobility in adjustment of the test to allow these changes to take place. It is also of interest in support of the view urged so much by Professor Hyatt, that changes are taking place constantly throughout the life of the individual. Centrechinus setosus is slow and very gradual in developing its characters. It is slow in acquiring genital pores and very slow relatively in having ocular plates enter the periproct. Up to 15 mm. in diameter no oculars have yet entered the periproct. In the series 20 to 25 mm. in diameter, a little more than half of the specimens still have all oculars exsert. When 35 to 40 mm. in diameter, which is about half-grown for the locality, the species character of I, V, [Vinsert is developed toa high point, 71 per cent, practically the maximum percentage; but at this age there are more speci- mens representing developing characters and fewer representing progressive characters than in older series. ‘The maximum of the species character and also the maximum of the progressive characters of I, V, IV, II, and all oculars insert, is seen in the largest specimens of 50 to 80 mm. diameter. The slow rate of development of Centrechinus as here shown (p. 156) is graphically brought out by comparing it with that of the development of Strongylo- centrotus drébachiensis given in the ‘‘ Phylogeny of the Echini,’’ page 142. Centrechinus is a relatively primitive (and geologically old) genus in the lowest suborder (the Aulodonta) of the Centrechinoida. On the other hand, S. drébachiensis is a highly accelerated species in the highest suborder (the Camarodonta) of the Centrechinoida. As Professor Hyatt showed fully in his studies of fossil Cephalopoda, primitive genera have a slow development and specialized genera havea highly accelerated development. I have shown the same thing in Pelecypoda where the primitive Avicula and Pecien havea relatively slow development, whereas the specialized Spondylus and Ostrea have an accelerated development. Another feature of note in the ocular arrangement of Centrechinus setosus is the fact that while ocular I or ocular V may be the first to enter the peri- Studies of Jamaica Echint. 159 proct, V is by far the more common, and inso far makes an approach to the character of the Cidaride where ocular V alone is frequently insert, but I alone is rarely insert. In higher groups of the Centrechinoida, when one ocular is insert, the order is more definitely fixed, as in the Arbaciidz and Echinometridz, where V is typically and I is rarely insert; or in the Echinidze and Strongylocentrotidz, where I is typically and Vis rarely insert. Another ocular peculiarity of Centrechinus is that where four plates reach the peri- proct, I, V, IV, III insert is a rather frequent aberrant variant. This is a very rare character in the higher Centrechinoida but is a common character in the Cidaride. Of the ocular characters of Toxopneustes variegatus, no special comment is called for. It is simply the record from a definite locality, whereas my previous record of the species was based on specimens from several locali- ties. The peculiar zigzag pattern of color ornamentation shown recalls the somewhat similar pattern seen in some of the Temnopleuridz. In Tripneustes esculentus the ocular arrangement (figs. 10-17) is one of the most interesting that has been found in any Echini. Two oculars insert is commonly given as the species character, and it is the dominant character in some localities (Bermuda, Florida). In Montego Bay this character is acquired up to 90 per cent in specimens 40 to 50 mm. in diam- eter, much less than half-grown. But in Montego Bay, as a local char- acter, the species has gone much further and passed through a phase in which oculars I, V, IV are insert like Bahama material, and finally takes on oculars I, V, IV, IL insert as the fully matured adult character. Itisan exceptionally interesting case as being the acquirement in late stages of growth asa typical feature of a character which is a normal progressive variant of its own species in other localities and also a typical advanced character in all Echini which have four oculars insert. Looking at the test of a nearly grown Tripneustes with its solid heavy plates, it is astonishing to realize that they are still plastic and capable of allowing the readjustment of parts indicated by the separation of genitals and traveling in of oculars so as to reach the periproct. Not less surprising is the power of the law of ocular develop- ment, that not any ocular, but a certain specific ocular, is the one that is selected by the laws of growth to change its relative position. I know of no more striking case of the definiteness of action in the growth of organisms. As stated, Echinometra lucunter is one of the very few species of Echini characterized by having one ocular insert; therefore a series showing its development is of interest. Four of the six species in the genus Echino- metra are characterized by having all oculars exsert as the species feature, and in these to have one ocular insert is a progressive variant. Also asa family character, all exsert is the leading feature. Therefore for the genus and for the family, Echinometra lucunter can be considered an advanced type as regards ocular arrangement. In accordance with this fact it is not strange, it might even be expected, that it would take on its advanced character of oculars insert at a somewhat late stage as it does. The domi- 160 Papers from the Marine Biological Laboratory at Tortugas. nance of V insert, 54 per cent, is first attained when the specimens are 25 to 30 mm. in diameter, about half grown, and the relative increase of V insert progresses somewhat up to the largest series recorded. Having oculars V, I insert is a progressive character for this species, and this feature comes in late, progressively increasing in frequency up to the largest size recorded. To have oculars V,I insert as a typical character is the feature of Echinometra van brunti A. Agassiz, from Lower California and South America, and in this respect it is the most specialized species in the genus and family. In the “ Phylogeny of the Echini’’ much attention was given to ocular plate arrangement and development of the same. The results for the Cent- rechinoida are there briefly summarized in pages 92-94. As observations on the Montego Bay series have materially extended the results in some important particulars, a brief statement of the laws of ocular arrangement is given here. In the “ Phylogeny”’ the results of tabulating ocular plates in young and adults of the order Centrechinoida are given for 48,541 specimens. Those given above include in addition 2,878 specimens, or including both, 51,419 specimens. By the law of ocular development as worked out in this order, the oculars when entering the periproct, do so in the sequence I, Vor V,I, IV, II, III (figs. 10-14), and it is seen by the following how closely this rule is adhered to. Of the total 51,419 specimens of Centrechinoida observed, oculars are all exsert in 6,763 specimens, all others having one or more oculars insert. In 3,708 specimens, one ocular is insert, and of these in 3,679 cases it is either ocular I (fig. 10) or ocular V, that is, 99.22 per cent are correct byrule. Thereare only 29 exceptions.! In 35,951 specimens two oculars are insert, and of these, in 35,611 cases, it is oculars I and V, or the bivium (fig. 11), that is, 99.05 per cent are correct by rule. In all of the 340 exceptions? one of the two platesinsert is eitherI or V. In 3,820 speci- mens, three oculars reach the periproct, and of these, in 3,426 cases, it is oculars I, V,IV, the bivium and the left posterior plate of the trivium (fig. 12), that is, 89.68 per cent are correct by rule. Of the 394 exceptions in 360 cases, the order is I, V, II insert (fig. 15), that is, the right posterior plate of the trivium is insert instead of the left posterior as usual. These may therefore be considered as right-handed specimens. Of the total 3,820 specimens with three oculars insert, 3,786, or 99.37 per cent, have either oculars I, V, IV, or oculars I, V, II insert. In 619 specimens, four oculars reach the periproct, and of these in 554 cases, it is oculars I, V, IV, II the bivium and posterior pair of the trivium (fig. 13); that is, 89.50 per cent are correct by rule.’ In 558 cases all five oculars are insert (fig. 14). Inthe oe eo ad ee, 18 have ocular IV alone insert, three have ocular III alone insert, and eight have 2 Of these exceptions, in ror cases, oculars I, IV are insert; in one case oculars I, III are insert; in 56 cases oculars I, II are insert, which is the species character in Gymnechinus robillardi (Loriol) and G. pulchellus geen to : 13 cases oculars V, IV are insert; in 4 cases oculars V, III are insert; and in 65 cases oculars , 3 Of the other 34 exceptions to the rule where 3 oculars are insert, in 4 cases oculars I, V, III are insert; An ue perhaps I, IV, II are insert; in 11 casesoculars V, IV, III are insert; and in 18 cases oculars V, 4 Of the 65 exceptions in 64 cases. the arrangement is oculars I, V, IV, III insert (fig. 16); this character which is rare in the Centrechinoida as a whole, occurred principally in Centrechinus setosus (47 cases), but it is a common character in the Cidaroida, the other 17 cases of I, V, IV, II insert are scattered through a number of species of the Centrechinoida. One aberrant with four oculars insert has oculars I, V, II, III reaching the periproct (fig. 17). This, which may be considered a right-handed equivalent of I, V, IV, III as far as experience goes, is a unique variant in Echini. Studies of Jamaica Echini. 161 “Phylogeny” a detailed discussion is given of the bearing of aberrant variants which, although aberrant, follow quite definite lines of their own. Of the total 51,419 specimens considered above, 50,591, or 98.39 per cent, are correct by rule as regards ocular arrangement, and 828 specimens (of which 360 are I, V, II as above noted), or 1.61 per cent, have an aberrant arrangement. From the above records it is seen that ocular plates in the Centrechinoida follow a very definite line as regards which ones reach the periproct, and by their arrangement emphasize a bilateral symmetry through the axis of ambulacrum and ocular III and interambulacrum and genital 5. This is the axis on which the irregular types develop an elongate form, eccentric periproct and other features of bilaterality, and it was urged by Lovén as the correct axis for orienting Echini. Mr. A. Agassiz (Challenger Echini, pp. 4-6) criticizes Lovén’s determination of the antero-posterior axis in Echini. His argument, however, simply shows that he misunderstood Lovén’s point of view. In accordance with what I previously showed in Strongylocentrotus it is seen that the perforation of the genital plates by the genital pores originates at quite definite periods. In Centrechinus setosus the genitals are imper- forate in specimens up to 14 mm. in diameter, after which with few ex- ceptions all specimens have developed the genital pores. In Echinometra, genitals are imperforate up to 10 mm., after which all plates are perforate. Both of these are later in developing than Strongylocentrotus drébachiensis, which is imperforate only up to 5 mm. in diameter, after which in most cases the genitals are perforate. In Eucidaris tribuloides and Centrechinus setosus the teeth extend to the upper line of the lantern, but do not extend horizontally over the top of the same; also, in both the dental capsules embracing the base of the teeth are small. These are both doubtless primitive conditions, and are opposed to the condition found in the Echinide and Strongylocentrotide. In Toxopneustes variegatus, Tripneustes esculentus, and Echinometra lucunter the teeth extend horizontally over the top of the lantern, the proximal end of the tooth is grooved, not keeled, the keel appearing further down the tooth, all as in Strongylocentrotus. This grooved character is of interest as a localized stage in development, as it shows that the young growing point of the tooth has the character retained throughout life in the Aulodonta, Cidaroida, and Palzozoic genera, in which the teeth are always grooved. In Toxopneustes, Tripneustes, and Echinometra the dental capsules are large, here again being like the character of Strongylocentrotus and differing from the condition in Eucidaris and Centrechinus. In Centrechinus setosus the perignathic girdle is specialized in the adult, with high apophyses and high laminar auricles joined over the ambulacra. In the young, the apophyses are very slight and the auricles are simple spur-like processes, as in the adult Aspidodiadematide, which represent the simplest auricles known. Later, in Centrechinus, the auricles arch over the ambulacra, but without fusion. Then they fuse in delicate arches, a It 162 Papers from the Marine Biological Laboratory at Tortugas. condition comparable to that of the adults of Chetodiadema pallidum A. Agassiz and Clark and Centrostephanus rodgerst A. Agassiz. Later still, in Centrechinus, the auricles develop laminar expansions on their upper border, which in large specimens become extensive. In Echinometra lucunter, the perignathic girdle is also very specialized, attaining an extravagant development of the auricles. In the young, however, the auricles exist as simple separate styles. Later they arch over the ambulacra, but without being in contact; still later they join over the ambulacra as delicate arches, a condition comparable to that seen as the adult character in simpler species of the genus. Finally, the auricles take on a great development of the dorsal border, producing the spoon-like arches characteristic of the species. As shown and urged by Hyatt from his studies of Cephalopoda, and as I have shown in previous studies, in the present study of Echini, and in plants, development, that is the addition of differential characters, con- tinues more or less markedly throughout the life of the individual, and a study of such development yields facts of value in the structure, the mor- phology, and the phylogenetic relations of the group in hand. X. THE SPERMATOGENESIS OF THE MONGOOSE; AND FURTHER COMPARATIVE STUDY OF MAMMALIAN SPERMATOGENESIS, WITH SPECIAL REFER- ENCE TO SEX CHROMOSOMES. By H. E. JORDAN, Professor of Histology and Embryology, University of Virginia. One plate and nine text-figures. THE SPERMATOGENESIS OF THE MONGOOSE; AND A FURTHER COM- PARATIVE STUDY OF MAMMALIAN SPERMATOGENESIS, WITH SPECIAL REFERENCE TO SEX CHROMOSOMES. By H. E. JorpAn. INTRODUCTORY. In view of the recent discovery of heterochromosomes in a number of the higher vertebrates it seemed desirable that the spermatogenesis of still other forms should be carefully studied with particular attention to the possible presence and variable behavior of homologous ‘‘accessory”’ chro- mosomal elements. While this study was in process opportunity was afforded to collect in Jamaica, British West Indies, material of the mon- goose.! This material is interesting in a number of respects and serves well as a starting-point for such comparative study as will appear from its description. Other material at hand includes that of cat, squirrel, pig, rabbit, white mouse, sheep, horse, mule, bull, and dog. HETEROCHROMOSOMES LACKING IN MALE. MONGOOSE. Spermatogonial nuclei more generally contain three plasmosomes (fig. 1), though there may be more or fewer. Only a very few nuclei were seen in mitosis. Many stages of apparent amitosis are present (fig. 2). The rarity of mitotic figures and of binucleate cells indicates that the pre- vailing type of division among the spermatogonia is amitotic. The presence, however, of a few binucleate spermatocytes and spermatids shows that cell-body divisions do not always follow the nuclear divisions. The primary spermatocytes contain a single nucleolus (plasmosome), seen only in the resting condition of the nucleus. The fact that this body never appears subsequently makes the synapsis and division stages of the mongoose especially favorable for study with reference to the presence of an “accessory chromosome.”’ There can be no confusion here between a plasmosome and a chromosome-nucleolus, as has apparently been the case in some forms. Moreover, the early disappearance of the plasmosome in mongoose gives support to the general conclusion that synaptic and post- synaptic chromatin elements are chromosomes, more especially when invariably on the nuclear membrane instead of being disposed at random. The synizesis phase is characterized by a very compact, deeply chro- matic mass of simple threads (fig. 4). The mass is situated at the pole of 1 This opportunity occurred during a month’s stay at the temporary Marine Biological Laboratory of the Carnegie Institution of Washington at Montego Bay in the spring of 1912. I take this occasion to express my thanks for the privileges of ay Institution’s expedition to Jamaica, and for daily kindnesses on the part of the director, Dr. Alfred G. M 165 166 Papers from the Marine Biological Laboratory at Tortugas. the nucleus, frequently, but not invariably, next the idiosome. The re- mainder of the nuclear space remains empty, except for a few delicate linin threads, more abundant in the vicinity of the mass. All about the idiosome are pale spherical mitochondria, now apparently present for the first time; but no clue appears as to their origin. The fact that the synizesis nucleus is not appreciably larger than that of the resting stage shows that synizesis is not due to mechanical factors consequent upon a great increase in nuclear sap. On the contrary, this phenomenon is unquestionably the result of the specific activity of the chromatic nuclear thread. Its frequently polar position may be due to idiosome influence. In the bouquet stage the simple threads emerge from the synizesis mass in pairs (fig. 5) and unite side by side (parasynapsis). The nucleus is still of approximately the same size; and again the chromatin threads give evi- dence of a specific activity. The process continues until the previously empty nucleus is filled with double threads crossing in every direction throughout the nucleus (fig. 6). Subsequently the nucleus enlarges some- what, and the double threads shorten and thicken (fig. 7). The beginning of division of the idiosome marks this as the early prophase. The bivalent chromosomes appear more chromatic and knobbed at the ends, an early indication of tetrad formation. Figure 8 illustrates a later prophase stage, in which the chromosomes have become still shorter, uniformly chromatic, and more compact, a number appearing as complete tetrads. The point of special interest is that nowhere throughout these early phases has the slightest evidence of an accessory chromosome appeared. No evidence would be expected to appear subsequently, and this is actually the case, as shown in figures 9 to 17. At metaphase the chromosomes are all arranged in a very compact equatorial plate (figs. 9 and 10). No chromosome seems marked by peculiar behavior or uncommon size. An accurate count seems impossible. Innumerable division figures appear, ‘but the chromosomes are so closely compacted as to preclude definition of limits. A most careful study warrants only the statement that the number of chromosomes is somewhere around 24. A resting stage ensues, but without chromosome-nucleolus, in contra- distinction to the usual condition in forms with an accessory chromosome, or without even a plasmosome. The division figures of the second mitosis are smaller, somewhat less compact, and seem to consist of fewer chromo- somes. We may be dealing here, asin the opossum, with a second pairing, or a hemioid group (Jordan, 1911). The smaller size of the chromosomes, however, makes this a little doubtful. Figure 14 illustrates a resting spermatid, with conspicuous idiosome. At the next step in the metamorphosis (fig. 15) the cell has elongated, the nucleus has become more dense and in consequence smaller, and the idio- some has given place to the archoplasmic sphere. The latter contains centrally a chromatic granule, presumably the centrosome (fig. 15). The Spermatogenesis of the Mongoose, etc. 167 sphere seems to have arisen directly from the idiosome by process of vesic- ulation and rejection of one or several small plastin particles. The con- densing nucleus moves into an elongating projection of the cell, and then to and beyond the periphery (figs. 16 and 17). Meanwhile the ‘‘sphere’’ has flowed backward over the elongating nucleus, its lateral limits being marked by a distinct line for some distance on either side of the growing axial fila- ment. The filament is marked proximally by two granules (the products of the original centrosome, fig. 15), the anterior one at the point of attach- ment to the nucleus, the posterior one passing along the axial filament as the ‘‘ring centrosome’”’ (figs. 17 and 18) to the posterior end of the middle piece. At the later and adult stages the middle piece is attached to the head by a delicate neck (fig. 20, drawn in the living condition), in which appears a knob, the proximal centrosome, or ‘‘end knob”’ (figs. 19 to 21). The posterior limit of the middle piece is defined by the termination of the spiral filament. The latter arises by a coalescence of mitochondria included within the limits of the backward-extending sphere (middle piece). The remaining mitochondria fail of inclusion and are cast off with the discarded cytoplasm. This observation, made also in a number of other forms (e. g., opossum, Jordan, 1911; Euchistus, Montgomery, 1912), invalidates 5 any hypothesis attributing specific hereditary significance to mitochondria. CAT. The later steps in the spermatogenesis of the cat, including the spermatid and subsequent stages, have recently been described by Leplat (1910). As to the prespermatid stages, it may be said in brief that they are essentially similar to those described for mongoose, and no convincing evidence appears at any stage of heterochromosomes. Respecting the postspermatid stages also, the similarity amounts practically to an identity; and my own obser- vations on these stages agree essentially with those made and illustrated by Leplat. More recently Gutherz (1912) has described and illustrated (in two figures) first spermatocytes of the cat stained both in the iron-hematoxylin and the Biondi mixtures. His conclusion confirms my own as to the probable absence of heterochromosomes. However, he figures a synaptic (synizesis) and a postsynaptic stage in which appear heterochromosome- like elements in iron-hematoxylin preparations. The true acidophile nature of these bodies, however, is revealed in the Biondi material. On the basis of this observation he casts doubt upon the interpretation as a heterochromosome of the element described by Winiwarter and Sainmont in their iron-hematoxylin preparations of the odcyte of the cat. I can only add that my material (also stained with iron-hematoxylin) reveals elements somewhat similar to those described and illustrated by Gutherz for the first spermatocytes, but which, on grounds of morphology, location, relation to spireme, and general behavior, I can not interpret as typical X-elements. Moreover, in well-decolorized preparations their non-chromatic nature is 168 Papers from the Marine Biological Laboratory at Tortugas. likewise revealed. The illustrations of the odcytes of the cat by Winiwarter and Sainmont (see figs. 27a, 33, and 44) answer to the main criterion, namely morphological, for the presence of a heterochromosome, ‘‘split monosome.”’ A comparison of the illustrations of the spermatocytes and oécytes of the cat as given by Gutherz and by Winiwarter and Sainmont respectively does not seem to warrant the conclusion of an homology between the hypo- thetical X-element in the male and that in female germ-cells of the cat. In short, the odcytes would seem to contain, the spermatocytes to lack, typical X-elements. SQUIRREL. The spermatogenesis of the squirrel has been described in two papers by Van Mollé (1906, 1907). The cells are relatively very large and the idio- some is very distinct. My observations agree with the illustrations of Van Mollé with respect to the absence of heterochromosomes. Duesberg (1910) has recently questioned the accuracy of Van Mollé’s observations with respect to the details of the later stages in the metamorphosis of the spermatid. Interest in these details is outside the scope of this contribution, and my observations on these points have not yet been carried to an extent where I might confidently presume to express an opinion. The absence of any accessory chromosomal element during the growth period appears un- questionable. PIG. It will suffice for present purposes to state simply that evidence of an accessory or heterotropic chromosome at early stages of the spermatogenesis of the pig is completely lacking in my material. The cells here are relatively small; but the fixation is good and reveals clearly all the details of structure, and every phase is abundantly represented prior to the division phases; oc- casionally first maturation groups are also present.! RABBIT. Exactly the same statement holds for the rabbit as for the pig; and I make it with greater confidence, since the material is more extensive including besides adult testes those of new-born and young individuals. In all of the foregoing instances the spermatogonia contain plasmosomes. The resting primary spermatocytes also contain either plasmosomes or karyosomes (net-knots), or both, all of which disappear in early presynapsis. It seems legitimate to assume, on the basis mainly of this observation, that in mammals generally plasmosomes almost invariably disappear before 1 However, no chromosome counts were attempted. In view of J. E. Wodsedalek’s recent work (‘‘Acces- sory Chromosomes in Pig;” Science, N.S., vol. xxxvi, No. 966, pp. 30-31) reporting two additional chro- mosomes or accessories in half of the secondary spermatocytes—this number (10) corresponding with the re- duced number in the eggs—the pig can no longer be listed under ‘‘exceptions.’’ In the absence of definite evidence during the growth period, the final criterion for the presence of heterochromosomes must be an actual count. Since the appearance of Wodsedalek’s complete paper (Biol. Bull., 25: 1), I have carefully re-examined my sections of the pigs’ testes. Mitoses are too infrequent to warrant any statement concerning the presence or absence of accessory chromosomes on the basis of appearances at metaphase, or of chromosome counts. My conclusions rested upon evidence drawn from a study of appearances during the growth stages including the early prophase. As to the resting spermatogonial and primary spermatocyte nuclei I find no such con- stancy with reference to nucleoli as Wodsedalek records. The number, both of the large and smaller nuc- leoli, is very variable. Three or four large nucleoli are common. Moreover, with reference to the chro- mosome-nucleoli I can not find in my preparations (of young testes) any cells to parallel his illustrations 21, 23, 24,and 25. My material does not confirm his finding that “two large round nucleoli remain very conspicuous throughout the process of growth of the primary spermatocyte.” Spermatogenesis of the Mongoose, etc. 169 synapsis, and can thus produce no confusion at those stages where hetero- chromosomes when present are most conspicuous, 7. e., synaptic and early postsynaptic stages. The conclusion that heterochromosomes are wanting in the male of the mongoose, cat, squirrel, pig, and rabbit is, of course, based on the assumption that if present they would be conspicuous at the same stages in which they are so strikingly in evidence in the group of mammals next to be described. The final test must, however, in these cases be an actual count of chromosomes. HETEROCHROMOSOMES PRESENT IN MALE. WHITE MOUSE. Spermatogonial and spermatocyte resting nuclei agree in being vesicular and having a delicate chromatic reticulum with numerous net-knots (figs. 22 and 23); they differ in that the spermatocyte nucleus has a bilobed or double chromatin (chromosome) nucleolus. The Sertoli cell has a still more vesicular nucleus, which is very con- spicuous by reason of a characteristic trilobed, deep-staining nucleolus, the central section being invariably the larger. This peculiar type of nucleolus is as characteristic of the Sertoli cell as its peculiar shape, and suggests interesting speculations concerning the relationship between nucleolar morphology and cell functions. My earlier observations led me to the conclusion that the synapsis phenomenon (i. e., synaptic knot, contraction or bouquet phase, or synizesis stage)! was lacking in the auxocytes of the white mouse. Further careful search has disclosed a few cells in the condition illustrated in figure 24. This is tentatively interpreted as synapsis (polarized amphitene). If the nucleus is normal, as it appears to be, the paired threads unmistakably indicate synapsis. I find that Regaud (1909, 1910) reports his failure to find a synapsis phase also in the rat. This phase is assuredly not accentu- ated in white mouse. Nothing more closely resembling synapsis than the nuclear arrangement illustrated in figure 24 could be found in my material. This, however, seems sufficiently suggestive of true synapsis to show that something similar to synapsis actually occurs in the mouse, perhaps too rapidly to appear except very occasionally in “‘fixed’’ preparations. If the phenomenon is really lacking the “‘synapsis”’ figure does not appear essen- tial to the pairing of the chromosomes in the formation of the haploid group. During synapsis (parasynapsis) and early postsynapsis (figs. 24 and 25) the chromatic accessory body (chromosome) is apparently usually single, usually oval, sometimes slightly bilobed, and occasionally wedge-shaped or even irregular. During later postsynaptic (fig. 26) and prophase (fig. 27) stages, this element again appears invariably double, frequently in the form of a longitudinally split rod. There is no unequivocal evidence that the 1 Synapsis is assumed to take place during the period of contraction or ‘‘ synizesis’; hence the terms are here used interchangeably. 170 Papers from the Marine Biological Laboratory at Tortugas. pair may entirely and widely separate, as is the case in the mule for example. The double nature of the heterochromosome suggests a pair of idiochro- mosomes; but the fact that during and immediately after synapsis this element appears single invalidates somewhat such an assumption, and suggests rather a double accessory chromosome, or X-element of Wilson. In leptotene and diplotene phases it very frequently lies in a clear space free of threads, a condition similar to that described by Wilson (1912) for Largus and Oncopeltus. Figure 27a would seem to permit of not the slightest doubt of the presence of a “‘sex chromosome”’ (Wilson) in the white mouse. Its later history, however, is still obscure. Division figures are very abundant in my material and the preservation is nothing short of superb, but at no later stage can I with certainty identify this body. Occasionally a spindle appears with a large chromosome in advance of the main metaphase group (fig. 27b), but the instances seem too rare to have much significance apart from morpho- logical marks of identification from among the chromosomes of the meta- phase plates. SHEEP. In sheep I am able to distinguish the ‘‘X-element’’ during the presyn- aptic (fig. 28), synaptic (fig. 29), and postsynaptic (fig. 30) stages. Both in the first and second stages it is usually attached to the chromatic spireme, or one of its segments. During the postsynaptic stages it is usually split or bilobed, and, as at earlier stages, frequently but not invariably lies close to the idiosome. Synapsis is effected by side-by-side union. In the sheep, also, this accessory chromosome eludes later certain identification. HORSE. As previously briefly noted (Jordan, 1911, 1912), during postsynaptic stages the horse shows frequently a tripartite X-element (fig. 33), usually next the idiosome. In the resting phase of the primary spermatocyte (fig. 31), and during synapsis, it appears double. Further clear tracing of this body seems impossible. The evidence is thus far from sufficient to warrant even an inference as to whether we are here dealing with a multiple X-element (as in Sinea, for example, according to Payne, 1909) or a pair of idiochromosomes. Both conditions have been described for mammals; for example, man as having a double X-element (as in Syromastes—Wilson, 1909) by Guyer (1910), and guinea-pig with a pair of heterochromosomes, by Miss Stevens (1912). The only point sought to establish is the presence of “sex chromosomes”’ of some sort. Kirillow (1912) has recently published the first number of his projected studies on the spermatogenesis of the horse. The accompanying illustra- tions, including all the chief stages, give no indication of a heterotropic body so conspicuous in my preparations; nor is any mention made of such body. His observations on the germ-cells of the horse lead him to accept the opinion of Regaud (1909, 1910) that ‘the appearance or failure of Spermatogenesis of the Mongoose, etc. 171 synapsis is a variable factor in different species”’ (Kirillow, p. 145); that is, synapsis (contraction phase) is regarded as a normal phase in the auxocytes of some forms (species), but is thought not to appear in others. MULE. The presence of an accessory chromosome in the mule was also noted briefly in an earlier paper (Jordan, 1911, 1912). Here it usually appears double in pre- and post-synaptic stages (figs. 34 and 36). At synapsis it appears as a single large oval chromatic body (fig. 35). It is very clearly recognizable among the prophase chromosomes as a large paired element (fig. 37). Since at certain phases (synapsis) it may be a single compact body, it is perhaps more legitimate to regard it as a double X-element. In the case of the mule it will be impossible further to trace this body with any degree of certainty. As first noted by myself (see quotation from my unpublished manuscript in a paper by Dr. R. H. Whitehead, ‘“‘A Peculiar Case of Cryporchism, and its Bearing upon the Problem of the Function of the Interstitial Cell of the Testis,” Anat. Record, Aug. 1908; vol. 11, No. 5), spermatogenesis in the mule does not ordinarily proceed beyond the early prophase. This is the cause of the sterility of mules bred inter se; the mule does not generally produce ripe spermatozoa. My obser- vations were subsequently confirmed by Poll (1911), who succeeded in observing a few first maturation spindles at metaphase. BULL. The spermatogenesis of the bull has been fully described by Schoenfeld (1901). My observations agree essentially with his illustrations. Schoen- feld figures a stage similar to my illustration of postsynapsis (fig. 38), where a single large, oval, chromatic “ accessory chromosome ”’ appears among the bivalent threads, close to the idiosome. This element, at this as well as at earlier and later stages, Schoenfeld describes as the ‘“‘corpuscle intra- nucleare.”” In the light of what we now know of ‘‘sex-chromosomes”’ generally, and in mammals more particularly, this body may, I believe, be regarded confidently as an accessory chromosome. DOG. Figure 39 illustrates a resting primary spermatocyte with pale plasmo- some and chromatic, bilobed chromosome nucleolus. No very satisfactory synapsis stages could be observed in my material.! Figures 40 and 41 illustrate two successive postsynaptic phases in which the X-element is conspicuously present as a chromatic, sharply contoured, irregularly oval body close to the idiosome. We are probably here again dealing with an accessory chromosome. ‘The attempt further to trace this body, however, was very disconcerting by reason of the absence of definite marks of identity, lack of clear delimitation of the relatively large number of small chromo- ‘somes, and confusing contradictions of trying observations. In short, any further analysis was unsuccessful. 1 That is, from the standpoint of a conspicuous ‘‘ X-element.” 172 Papers from the Marine Biological Laboratory at Tortugas. DISCUSSION. Heterochromosomes are apparently lacking in mongoose, cat, squirrel, pig, and rabbit, and appear to be present in white mouse, sheep, horse, mule, bull, and dog. On the basis of what was already known regard- ing man (Guyer, 1910; Gutherz, 1912),! rat (Guyer, I910), armadillo (Newman and Patterson, 1910), guinea-pig (Stevens, I912), and opossum and bat (Jordan, 1911, 1912), heterochromosomes were suspected in mam- mals generally. With respect to heterochromosomes, then, mammals fall into two groups: those lacking and those possessing the X-element in the male germ-cells. Under the first group five? mammals can be classified; under the second, twelve. Both groups include higher as well as lower mammals. Measured by no other criterion known to me can these mammals be similarly grouped. Presence of heterochromosomes obviously bears no relationship to natural affinities or evolutionary levels of mammals. The proportion of 12 to 5 among examined and recorded cases of mammals in favor of those possessing heterochromosomes suggests that in those forms apparently lacking them they have simply eluded detection. They might presumably be so small as to escape recognition, or lack the peculiar morphological marks by which they are usually characterized, or be too labile to be identified. Regarding a similar condition in Culex, Stevens (1911, 1912) suggests that here, “‘where no heterochromosome differentiation of any kind has been detected, the members of a pair of chromosomes may differ by a sex unit or some other character unit, and the difference in size comes within the probable error of most careful observation”’ (p. 165). The observation of the attachment of the accessory chromosome to the parasynaptic spireme (figs. 28, 29, and 40) controverts the plausibility of Stevens’s (1912) suggestion that ‘‘the condensed condition of the odd chro- mosome and the unequally paired heterochromosomes of the growth stage of the first spermatocytes might be due to their unpaired or unequally paired condition preventing them from joining with the other bivalent chromo- somes to form a spireme, especially in cases of parasynapsis”’ (p. 166). It would seem to be required, then, for a proper grounding of later hypotheses, that all uncertainty be disposed of, first with respect to the possible presence in some form or at some stage of an X-element in those animals in which it seems to be wanting; and second, with respect to possible confusion with a persisting plasmosome in those instances where an X- element is believed to obtain. Though the above order would seem to be the more logical, the best approach to the argument and the proof is per- haps by way of the second point, though in fact they sustain a reciprocal relationship. 1 Gutherz (1912) confirms Guyer’s (1910) observation respecting the presence of a heterotropic element in man, but interprets it as a pair of equal idiochromosomes undergoing neither heterokinesis nor giving rise to a dimorphism of spermatozoa as described by Guyer. 4 f : 2 This must be a tentative grouping; only four mammals can now be temporarily placed in this group. Spermatogenesis of the Mongoose, etc. 73 Regarding a possible confusion between plasmosomes and accessory chromosomes, or X-elements, it must be noted first that in the stain em- ployed (Heidenhain’s iron-hematoxylin; checked in doubtful instances by Auerbach’s methyl-green-acid-fuchsin stain) the plasmosome when present (as in spermatogonia and occasionally in primary spermatocytes) is always in properly decolorized preparations considerably less deeply stained than the X-element. Moreover, the latter is frequently attached to the spireme or one of its segments, and generally holds a position close to the nuclear wall and close to the point where the idiosome lies. Again, it is frequently bilobed or compound. In certain instances, e. g., mouse more particularly, one of the first-division chromosomes occasionally passes to the pole in advance of the mass; but in no instance could the X-element be followed with any satisfaction or certainty beyond the early prophase. At this stage, however, it is in at least a number of instances clearly recognizable among the pale, mossy chromosomes as the body earlier identified as the X-element. The evidence would seem amply adequate to support the interpretation here as a typical X-element. Heterotropic nuclear elements must be identified by other criteria than staining reaction. At best, staining capacity can only give confirmatory evidence. Morphological marks would seem to be the most certain grounds for basing distinctions. The complete nuclear history is of course necessary for full certainty. Chromosomes undergo alterations in their chromaticity and consequent staining capacity at different phases of the nuclear cycle. Again, true chromatin-nucleoli occur in certain forms (e. g., odcytes of Asterias forbesii, Jordan, 1908; odcytes of Echinaster and Cribrella, Jordan, 1910), whose function at least in part is to contribute chromatic material to the chromosomes just before they enter the first maturation spindle. The plasmosomes of certain forms may thus be of true chromatic nature. Moreover, true nucleoli never assume—except perhaps very exceptionally and atypically—the characteristic bilobed and split forms of the chromo- some-nucleoli during the growth stages. Degenerating nucleoli become ragged of outline, frequently fragment, and undergo karyolysis, but never show the series of phenomena characteristic of heterochromosomes: sharp contour, deep basophily, frequently split form, frequent attachment to spireme, and usual location close to the nuclear membrane. A presumptive chromosome-nucleolus may meet the test of basophilic staining reaction in a differential dye, but if it does not meet at least the majority of the above structural and spatial tests it very probably is simply a chromatic plasmo- some; on the other hand, if it answer to the latter tests, confirmatory—but perhaps not crucial, in view of the undoubted chemical changes which chromosomes undergo—evidence accrues from a chromatic reaction to a “specific ’’ stain. Regarding the first stated point of possible doubt, then, nothing further can be said than that in the five forms enumerated as lacking an X-element, the tissue was similarly well preserved and stained, but notwithstanding 174 Papers from the Marine Biological Laboratory at Tortugas. careful search, not the slightest evidence of such an element in its usual morphological form and peculiar behavior with reference to spireme and idiosome could at any stage be detected; but this negative evidence obviously can not remove all uncertainty with respect to this first point. Heterochromosomes (allosomes, Montgomery) in mongoose, and other forms showing like growth-stage conditions, may possibly have disappeared by close union with one of the ordinary chromosomes and thus forced into the usual behavior of the autosomes during auxocyte phases. Transition stages in such process of disappearance are apparently shown in certain orthoptera (McClung, 1905) and amphibia (e. g., Necturus, King, 1912). Militating further against the objection of mistaken identity is the fact that in this group the plasmosome has almost invariably disappeared! before synapsis, when the X-element first exhibits its characteristic behavior. If plasmosome could be confused with X-element in mammals, a group of five, with no relatively closer affinity than obtains among the contrasted group, could hardly be expected to give consistent and unequivocal evidence in favor of complete absence. Moreover, if the plasmosome generally disap- pears before or about the time of synapsis in a relatively no more closely related group of five forms, it seems quite legitimate to infer that it disap- pears at about the same time in six other—and as closely related—forms, and thus leaves little opportunity for confusion. And the above deduction is supported to some extent by direct observations (exception: man). Having disposed of these objections as far as possible within the limita- tions of microchemical technic, it remains to consider further bearings and implications of the facts established. The observation of an X-element in the ovary of young cats (fig. 42) by Winiwarter and Sainmont (1909) has important significance in this connection. If my negative evidence for the male germ-cells in the cat can be regarded as equally certain with the positive evidence of these investigators for the female germ-cells, then it would seem that in those forms in which the males lack a typical X-element this is possessed by the female.?_ An analogous instance from the inverte- _ brate group is that of certain sea-urchins reported by Baltzer (1909). Here the female possesses a pair of idiochromosomes, while the male lacks any indication of such. The above hypothesis for mammals must be tested by examination of the young ovaries of those forms in which the male appears to lack the accessory. This hypothesis would be somewhat weakened if cases were known in which both male and female germ-cells possessed typical? X-elements. The meager evidence available is contrary to such a hypothetical condition. Wilson (1905, 1906) was unable to find a chromosome nucleolus in certain genera among the insects, including Euchistus, in which he was confirmed by Foote and Strobell (1909). Buchner’s (1909) evidence with respect to Gryllus—the only presumptive contradictory evidence known to me— 1A delayed disappearance occasionally occurs, more especially in cat and man. 2 Presumably by approximately half of the ova. 3 That is from the standpoint of behavior during the growth stages. Spermatogenesis of ihe Mongoose, etc. 175 judged from his illustrations, seems far from conclusive. In fact, the hypothetical so-called X-element (‘‘accessory body”’) in the ovary lacks much of being similar in structure and behavior to the undoubted X-element in Gtdipus (described and figured in the same paper) and the male Gryllus. Moreover, Gutherz (1910) has shown that the two X-chromosomes, char- acteristic of the female, are both present in the metaphase plates of the odgonia, in addition to the outlying ‘‘accessory body.” As is now estab- lished beyond any doubt the accessory, for example, of the male germ-cells passes into all females. Its homologue is present in both males and females; but the additional male accessory, at any rate, would seem to be able again to comport itself during the growth stage of the maturing oGcyte as it originally behaved in the growing spermatocyte; and similarly with respect to its female homologue, since that in the male, coming originally also from the egg, behaves again in typical ‘‘accessory’’ fashion. It seems fairly well established that the female homologues of the male accessory do not behave in the growing odcytes as does the X-element in the spermatocyte. As indicated by observation on the cat, the reverse of the usual relation- ship here obtains: the odcytes are probably dimorphic with respect to an accessory; the spermatocytes apparently monomorphic. Here, then, as in certain sea-urchins, the female is apparently digametic, the male apparently homogametic. Further investigations must establish the fact whether the same reverse condition obtains with respect to an X-element in the case of the remaining four mammals of the first group. Male and female sex (together with secondary sexual characters) appear to be hereditary in a manner similar to other organic characters, and may be regarded as a pair of ‘‘unit characters.’’ Since the femaleness is associ- ated with, or determined by, the presence of X-elements, the female (con- taining a pair of homologous X-elements) is presumably homozygous,! the male heterozygous for sex; or, in accordance with a more recent termin- ology, the female represents a duplex (but apparently recessive like a nulli- plex) condition, the male a simplex (dominant) condition. Maleness is accordingly characterized by the absence, femaleness by the presence of a “determiner’’ (presumably an inhibitor), the X-element. The former is the negative or minus (lacks accessory, contributed to female at fertili- zation), the latter the positive or plus, condition. Sex inheritance con- forms thus to the Mendelian scheme, in which maleness represents the DR and femaleness the RR condition; or, stated in terms of the presence (+) and absence (—) of an inhibiting accessory, or X-element, maleness is + —, and femaleness the +-+ condition. Femaleness may therefore be plus or positive in an inhibiting sense; a germinal plus condition prevailing in the female, inhibiting the appearance of a positive somatic condition or male sex. This modified Mendelian interpretation is suggested in part by an analogy with conditions in crosses between horned and hornless cattle, where the positive character (horns) is recessive to the negative or hornless 1 That is, in the group where the male germ-cells contain an X-element. 176 Papers from the Marine Biological Laboratory at Tortugas. character; meaning, according to the general interpretation, that hornless- ness is determined by the presence of some determiner which prevents the development of horns. Similarly, femaleness may be due to the presence of the X-element, preventing the development of maleness.! It would be immaterial whether it were contributed by the male or by the female gamete. This idea is further suggested by, and seems in perfect accord with, the numerous observations indicating that femaleness is undeveloped maleness, e. g., embryological facts; relatively female characteristics of male infants; the assumption of male secondary characters after spaying, or after the menopause; etc. SUMMARY AND CONCLUSIONS. Among the forms examined, including mongoose, cat, squirrel, pig, rabbit, white mouse, sheep, horse, mule, bull, and dog, heterochromosomes are apparently lacking in the male germ-cells of the first five, and present in the remainder. The available evidence favors more the interpretation in terms of a bipartite univalent or compound X-element than of an asso- ciated X and Y group (idiochromosomes). In view of the fact that heterochromosomes have recently been reported in man (Guyer, double X-element; Gutherz, equal pair of idiochromosomes, or X- and Y-elements), rat (Guyer), armadillo (Newman and Patterson), guinea-pig (Stevens), and opossum and bat (Jordan), the evidence indicating similar elements in the above-enumerated group of six common mammals would seem to warrant the conclusion that sex-chromosomes are very generally present in mammals. Universality of presence seems vitiated for the present by the fact that in another group of five mammals, carefully studied, such elements seem apparently lacking. It might be assumed that such elements are actually present in the male germ-cells, but are so small or labile as to elude detection by ordinary methods, or do not present the usual morphology of heterochromosomes during the prophase stages. The unmistakable presence, however, of a “‘split-accessory” in the female germ-cells (primary odcyte) of the cat, as recorded by Winiwarter and Sain- mont, and the absence of any X-element in the male (confirmed by Gutherz, 1912); suggests very forcibly that sex-chromosomes are present in all mammals, generally in the male, exceptionally in the female. The same result would follow (that of numerical sex-equality) whether present in one or the other sex. If this hypothesis can be further sustained, it would seem cogently to reinforce the evidence for an essential sex-determining function of heterochromosomes. Interpreted in terms of Mendelian heredity-formule, in those mammals in which an X-element is present in the male, the female sex is homozygous, the male heterozygous. The facts would seem to fit the hypothesis that the accessory chromosome acts as a deterrent to the develop- 1 However, in the case of horn-heredity, the inhibitor, according to this explanation, is dominant; in sex- heredity, recessive. In cases where the female is the heterozygote, the condition might be parallel to that of hormn-heredity. In these instances the female would be simplex, the male nulliplex. Recession of a duplex Sea ie seems a contradiction in terms. The suggestion can have significance in only an approximate or general sense. Spermatogenesis of the Mongoose, etc. E77 ment of maleness; or more accurately, and in keeping with a quantitative interpretation of sex in a final analysis, the accessory with its egg-homologue (two X-elements) inhibits male-sex development, the single egg-homologue in males being insufficient to counteract the male tendency, thus giving origin to male individuals. In those instances where the female can be shown to be the heterozygote, one X-element, according to the hypothesis, may be assumed to be able to counteract the male tendency. ADDENDUM. The opportunity recently presented itself for the study of very favorable human material. I am indebted to Dr. H. T. Marshall, professor of pathology at the University of Virginia, for the specimen, a thin slice of testicle fixed in Zenker’s fluid, and obtained at autopsy from a negro aged 38 years, who died as the result of drinking wood alcohol. The material is in excellent state of preservation. It was stained for study with iron- hematoxylin. Primary spermatocytes in early postsynaptic phases (text- figs. I to 9) are especially abundant and clear. Mitoses, however, both in spermatogonia and spermatocytes, are infrequent, and, though a sufficient number can be found at metaphase in the latter cells to furnish a fairly satisfactory opportunity for attempting chromosome counts, I can come to no definite conclusion concerning the actual number. My specimens of primary spermatocyte mitoses, however, compare very favorably with the illustrations given by Guyer and by Gutherz, every one of which I can practically duplicate. An impartial judge of the figures must admit, I believe, that certainty is impossible at least within an error of two—and two makes all the difference between the presence of an accessory as urged by Guyer and denied by Gutherz. The inherent difficulty of an accurate count is indicated by the fact that the diploid chromosome number of the male cells has been given as 22 (Guyer), 24 (Flemming, Duesberg, Branca, Gutherz), 16 (Bardeleben), 18 (Wilcox), 33 or 34 (Wieman), and 47 (Winiwarter). It is significant, however, that Guyer, Gutherz, Duesberg, and Branca agree on 12 as the haploid (reduced) number; but Guyer, in contrast to the others, regards 2 of these as univalent accessories; also Guyer, Gutherz, and Winiwarter agree in claiming the presence of a heterochromosome; and Guyer and Winiwarter both report a dimorphism of spermatozoa, the former giving the number of chromosomes as 10 and 12, and the latter as 23 and 24. It seems clear that a heterochromosome of some valency is present; it is equally clear that the true nature of this body can not yet be established on the basis of numerical relationships; the count is still uncertain as regards at least 2 chromosomes. “Study of my material convinces me that the reduced number is not less than 12, but perhaps several more. As regards conclusions respecting the presence of an accessory chromosome drawn from a study of side views of metaphase spindles, it may be pointed out that Guyer’s and Gutherz’s figures are substantially alike; but Guyer interprets I2 178 Papers from the Marine Biological Laboratory at Tortugas. the appearance as demonstrating, Gutherz as contradicting, the presence of an accessory. My own figures occasionally show a double chromosome at one pole in advance of the main complex, and the evidence favors Guyer’s interpretation rather than Gutherz’s, but there is here nothing so striking as one sees in certain insects (e. g., the phasmid, Aplopus mayeri) and the opossum. Strong evidence for the presence of a heterochromosome in man is given by Gutherz in his illustrations of early postsynaptic nuclei (text-figs. 2 to 6). But neither he nor Guyer seems to offer satisfactory evidence as to its composition from chromosome counts and behavior at metaphase. Both, moreover, regard it as a double structure. My own evidence confirms the findings of Guyer and Gutherz in so far as pertains to the actual presence and bipartite nature of a heterochromosome or X-ele- ment, and it emphasizes the additional points of similarity between this Fig. 1—Human primary spermatocyte; nucleus in early diplotene post-synaptic phase, showing a deeply staining oval chromatin (chromosome) nucleolus on the nuclear wall and attached to one of the threads; the true nucleolus is a spherical, less deeply staining body, and only very rarely placed close to the nuclear wall The cytoplasm is homogeneous, finely granular. Figs. 2 to 9.—Outlines of nuclei at same stage to show the general form and location of the heterochro- mosome. It is almost invariably bipartite and attached to one of the threads, and on the nuclear membrane. The nucleolus when present is indicated in outline. and undoubted sex-chromosomes, namely, frequent connection with the diplotene thread and close spatial relationship to nuclear wall (exception, text-fig. 3). Judging from the illustrations, Winiwarter (1912) has had the advantage of possessing by far the best human material. His illustration (fig. 25) is practically a duplicate of my own of the postsynaptic diplotene (passing into the pachytene) phase. However, the later history of the accessory element as traced by Winiwarter is not at all stages perfectly clear. But figures 39 and 40 show pairs of sister spermatids, one pair with, the other without, a deep-staining nuclear body (accessory chromosome?). If this spermatid element actually represents, as seems very probable, the postsyn- aptic nuclear bipartite body, or heterochromosome, then cogent additional evidence here also accrues favoring the legitimate interpretation as a heterochromosome of the similar (structurally and tinctorially) peculiar postsynaptic nuclear element wherever found in mammals. LITERATURE. BaLtTzeR, F. 1909. Die Chromosomen von Strongylocentrotus lividus und Echinus micro- tuberculatus. Arch. f. Zellf., Bd. 1, Heft 4. Bucuner, P. 1909. Das accessorische Chromosome in Spermatogenese und ovogenese der Orthoptern, zugleich ein Beitrag zur Kenntnis der Reduktion. Arch. f. Zellf., Bd. rv, Heft 2. DUESBERG, J. 1910. Nouvelles recherches sur l’appereil mitochondrial des cellules séminales. Arch. f. Zellf., Bd. v1, Heft 1. Foot and STROBELL. 1909. The nucleoli in the spermatocytes and germinal vesicles of Euchistus variclatus. Biol. Bull., vol. xv1, No. 5. Gross, J. 1912. Heterochromosomes and sex-determination. Zool. Jahrb., vol. Xxx. GUTHERZ, S. 1909. Wird die Annahme einer Beziehung zwischen Heterochromosomen und Geschlechts bestimmung durch das Studium der Gryllus-oogenese wiederlegt? Sitzungsber. d. Gesellsch. Naturforsch, Freunde, Nr. 9. 1912. Ueber ein bemerkenswertes Structurelement (Heterechromosom?) in der Spermiogenese des Menschen. Arch. f. Mikr. Anat., Abteilung 11, 79, 2. Guyer, M.F. t1g10. Accessory chromosomes in man. Biol. Bull., vol. xrx, No. 4. Jorpan, H. E. 1908. The spermatogenesis of Aplopus mayeri. Carnegie Institution of Washington Pub. No. 102. Abstract in Anat. Anz., vol. XXXII. — 1908. The relation of the nucleolus to the chromosomes in the primary oocyte of Asterias forbesii. Carnegie Institution of Washington Pub. No. 102. ——— 1g10. The relation of nucleoli to chromosomes in the egg of Cribrella sanguine- olenta Liitken. Arch. f. Zellf., Bd. v, Heft 3. ——— 1g911. The spermatogenesis of the opossum (Didelphys virginiana), with specia reference to the accessory chromosomes and the chondriosomes. Arch. f. Zellf., Bd. vi, Heft 1. 1912. Notes on the spermatogenesis of the bat. Anat. Anz., Bd. XL. Kinc, HELEN DEAN. 1912. Dimorphism in the spermatozoa of Necturus maculosus. Anat. Record, vol. v1, No. 10. Krrittow, S. 1912. Die Spermiogenese beim Pferde. I. Arch. f. Mikr. Anat., Bd. LxxIx, Abteilung 0, Heft 3. LEPLAT, G. I9Q10. i Spermiogénése chez le Chat. Arch. de Biol., Tome xxv, Fasc. 2 and 3. McCiunc, C. E. 1905. The chromosome complex of orthopteran spermatocytes. Biol. Bull., vol. 1x. Mo té, J. vAN. 1906. La Spermiogénése dans l’écureuil. La Cell. Tome xxi, Fasc. I. 1907. Les Spermatocytes dans l’écureuil. Ibid. Tome xxIv, Fasc. 2. Montcomery, T. H. 1910. On the dimegalous sperm and chromosomal variation of Euchistus, with reference to chromosomal continuity. Arch. f. Zellf., Bd. v, Heft 1. Morean, T. H. 1909. A biological and cytological study of sex determination in phylloxerans and aphids. Jour. Exp. Zool., vol. vir, No. 2. Newnan, H. H., and J. T. PATTERSON. 1910. The development of the nine-banded armadillo from the primitive streak to birth, with special reference to the question of specific polyembryony. Jour. Morph., vol. xx1, No. 3. Payne, F. 1909. Some new types of chromosome distribution and their relations to sex. Biol. Bull., vol. xvi, Nos. 3 and 4. Pott, H. 1911. Mischlings Studien V: Vorsamenbildung die Mischlingen. Arch. f. Mikr. Anat., Abteilung 1, Bd. Lxxvit, Heft 2. Recaup, C. 1909-1910. Etudes sur la Structure des Tubes Séminiféres et sur la Sper- matogénése chez les Mammiféres. Arch. de Anat. Micr., Tome x1. Paris. SCHOENFELD, H. 1902. La Spermatogénése chez le taureau, et chez les Mammiféres en général. Arch. de Biol., Tome xvi, Fasc. I. StEvENs, N. M. 1909. An unpaired heterochromosome in the aphids. Jour. Exp. Zool., vol. vi, No. I. 1911. Further studies on heterochromosomes in mosquitoes. Biol. Bull., vol. xx. 1912. Heterochromosomes in the guinea-pig. Biol. Bull., vol. xx1, No. 3. Wireman, H. L. 1913. Chromosomes in man. Am. Journ. Anat. 14: 4. Witson, E.B. 19054. The chromosomes in relation to the determination of sex. Science, n. s., vol. XX, 564. 1905B. The behavior of the idiochromosomes in Hemiptera. Studies on Chromo- somes, I. Jour. Exp. Zool., vol. 11, No. 3. ——— 1905c. The paired microchromosomes, idicchromosomes, and heterotrophic chromosomes in Hemiptera. Studies on Chromosomes, II. Ibid., vol. 11, No. 4. 179 180 Papers from the Marine Biological Laboratory at Tortugas. WILsoN, E. B. IQOQA. IQOQB. 1910. IQIl. 1912. 1906. The sexual differences of the chromosome groups in Hemiptera, with some considerations on the determination and heredity of sex. Studies on Chromosomes, III. Ibid., vol. 111, No. 1. The accessory chromosome in Syromastes and Pyrrhocoris with a com- parative review of the types of sexual differences of the chromosome groups. Studies on Chromosomes, IV. Ibid., vol. v1, No. 1. The chromosomes of Metapodius. A contribution to the hypothesis of the genetic continuity of chromosomes. Studies on Chromosomes, V. Ibid., vol. v1, No. 2. The chromosomes in relation to the determination of sex. Science Prog- ress, vol. Iv, No. 16. The sex chromosomes. Arch. f. Mikr. Anat., Bd. Lxxvu, Abteilung u, Heft 2. Observations on the maturation phenomena in certain Hemiptera and other forms, with considerations on synapsis and reduction. Studies on Chromosomes, VIII. Ibid., vol. x1m, No. 3. Von WINrIWARTER, H. 1912. Etudes sur la spermatogenése humaine. Arch. Biol. Tome XXVII. Von WInrtwartTer, H., and G. SAINMONT. 1909. Nouvelles recherches sur l’ovagénése et lVorganogénése de l’ovaire des Mammiféres (chat). Arch. Biol., Tome XxIVv, Fasc. 2 and 3. . Spermatogonium; DESCRIPTION ‘OF PLATE. Fixation: Flemming’s strong solution. MONGOOSE. the nucleus contains three small plasmosomes. . Spermatogonium; the nucleus is in process of amitotic division. Primary spermatocyte; the nucleus contains a centrally located plasmosome; the delicate chromatic reticulum consists of an apparently continuous single thread; leptotene-nucleus. Bouquet stage; the slightly thickened spireme has aggregated in a close-meshed mass at the idiosome pole of the nucleus; the cytoplasm surrounding the idiosome contains pale mito- chondria; synizesis, polarized amphitene. . Synapsis: the disentangling threads are pairing side by side; the nucleus is still approximately of the same size as in the resting spermatocyte, hence synizesis and synapsis signify activity (motion) on the part of the nuclear reticulum; synaptene-nucleus. . Postsynapsis; early prophase; diplotene-nucleus. and 8. Later prophases; at no stage do hetero- chromosomes appear. and to. Side and polar views respectively of metaphase plates; no chromosome is conspic- uous for unusual size or behavior; the number of chromosomes is approximately twenty- four, the haploid group. 1. Resting secondary spermatocyte. and 13. Side and polar views respectively of metaphase plates; the number of chromosomes is approximately twelve, forming a hemioid group. . Spermatid; the cytoplasm has become filled with pale mitochondria. . Appearance of ‘‘sphere’’ and centrosome. and 17. Appearance of axial filament and middle piece. and 19. Formation of spiral filament by coales- cence of mitochondria. _ and 21. Face and profile views respectively of mature, living spermatozoa. WHITE MOUSE. 22. Spermatogonium. 23. Primary spermatocyte, . Synapsis, nucleus at leptotene phase; the bilobed nucleolus may be a pair of heterochromosomes. polarized amphitene, bouquet or synaptene phase; the heterochromosome is apparently single. . Postsynapsis; early prophase; heterochrome (or accessory) conspicuous; pachytene-nucleus. and 27a. Early and late prophase groups respec- tively, both including conspicuously the pair of heterochromosomes almost invariably situated on the nuclear wall and close to the idiosome; 26, diplotene-nucleus. i t Stain: Heidenhain's iron hematoxylin. Magnification: 1500 diameters. 27b. Metaphase; one of the chromosomes has moved 20. 30. Bite oe 33. 34 35 36 38. . Resting primary spermatocyte, in advance of the others to one pole; this has a tetrad form and is still clearly bivalent; ac- cordingly, it fails to divide in this mitosis and may represent the ‘‘accessory’’ of the growth stages. SHEEP. showing the accessory at the idiosome pole; perhaps post- synaptic ‘‘confused stage’’ of Wilson. Bouquet (synapsis) stage; the accessory chromo- some is attached to a double thread, and in- variably lies close to the nuclear wall. Postsynapsis; early prophase, showing a bipartite accessory (or pair of heterochromosomes). HORSE. Primary spermatocyte; the nucleus contains two irregular karyosomes and a_ bilobed chromatic nucleolus, probably a pair of hetero- chromosomes. Synapsis stage, showing the heterochromosomes sues the pairing threads close to the nuclear wall. Postsynapsis, showing a tripartite X-element, the heterochromosome group; pachytene- nucleus. MULE. . Resting primary spermatocyte, showing a divided, or paired, heterochromosome. . Synapsis stage, showing a heterochromosome. and 37. Early and late prophase stages respec- tively, showing a pair of heterochromosomes. BULL. Postsynapsis, showing an accessory chromosome among the bivalent threads, close to wall and idiosome; diplotene-nucleus. Doc. . Primary spermatocyte; presynaptic resting phase, showing plasmosome and_ bilobed monosome. . Postsynaptic phase, pachytene-nucleus; mono- some attached to one of the threads. . Early prophase, diplotene-nucleus; chromosomes in form of long bivalent threads; monosome close to nuclear wall. . Odcyte from ovary of 23-day cat. Synapsis. True nucleolus; monosome divided longitu- dinally (after H. von Winiwarter et G. Sain- mont, fig. 43). JORDAN XI. THE BRYOZOA OF THE TORTUGAS ISLANDS, FLORIDA. BY RAYMOND C. OSBURN. Twenty-three text-figures. THE BRYOZOA OF THE TORTUGAS ISLANDS, FLORIDA. By Raymonp C. OsBURN. In the summer of 1908 the writer had the privilege of spending the month of June at the Carnegie Institution Laboratory for Marine Biology, located on Loggerhead Key of the Tortugas Islands. Owing to the short time at my disposal the entire period was devoted to a close search for the bryozoa inhabiting the shallow waters about the reefs, on the piles of the old government dock on Garden Key, in the moat of old Fort Jefferson on the same key, and in dredging the shallow waters about the islands down to 22 fathoms. For much of the work a skiff or a small launch was used, and for the deeper dredging (10 to 22 fathoms) the schooner Physalia was employed. The Tortugas Islands are islets of a coral atoll and in the diversified bottom afforded by such a region a rather abundant bryozoa fauna was found. Comparatively little work has been done on the bryozoa of the Florida and West Indian regions. Smitt’s papers on the Florida bryozoa (1872-73) deal with 87 species collected by Count L. F. de Pourtales and afford the only extended record of the bryozoa of this region. Pourtales (1867), in a paper entitled ‘‘ Distribution of the Fauna of the Gulf Stream at Great Depths,” listed and described as new 7 species of bryozoa, 2 of which have been proved to besynonyms. Verrill, in his Bermuda papers, has listed 21 species from the Bermuda Islands. Levinsen, in his important work, ‘Morphological and Systematic Studies on the Cheilostomatous Bryozoa” (1909), records 6 species from the West Indian and Florida region, describing two of them as new. Otherwise this vast region remains untouched. As the collections of Pourtales were made in the deeper waters of the Florida region, it was presumed that careful collecting in the shallow waters would disclose the presence of a somewhat different fauna, a presumption well borne out by the results of my collecting. There is, indeed, a remarkable disparity between the list given by Smitt and that forming the basis of the present paper. Not only did 41 of the species described by Smitt fail to appear in the shallow waters, but 40 others, whose presence was not hitherto suspected in the Florida region, and many of them not even in America, were taken. This brings the list of the species at present known from Florida and West Indian waters up to 127. By comparing with lists from other regions where the bryozoa have been carefully worked, it will be seen that the bryozoa fauna of the Tortugas and of the Florida-West Indian regions is fairly rich in species and fairly representative of tropical and semi-tropical regions. Careful collecting for 183 184 Papers from the Marine Biological Laboratory at Tortugas. a number of years in a limited region about Woods Hole, Massachusetts, and extending down to about 20 fathoms, yielded 83 species (see Osburn, 1912). Waters (1909-10) listed 73 species from the Red Sea. Norman (1909) records 139 species from Madeira and neighboring islands. This latter list is of special interest for comparison, since the Madeira bryozoa have been collected with some care for about 50 years and it may be supposed that it is fairly complete. In making comparison, however, it must be noted that the dredging about Madeira extended down to 200 to 300 fathoms, while those of the present list go only to 22 fathoms, so that to properly compare the Florida with the Madeira fauna the present list of shallow- water forms should be combined with that of Smitt. The Tortugas bryozoan fauna (shallow water) presents a very different facies from that of Woods Hole at the same depth, since the two lists of 76 and 83 species, respectively, have only 13 species in common. The Woods Hole fauna is distinctly northern, while that of the Tortugas, at least for the shallow waters, is distinctly tropical. This is readily understood when we recall that the Tortugas are within 0.5 degree of the tropics, and that these islands receive, directly along their shores, the warm waters of the Gulf Stream which sweep up from the Caribbean Sea. It is interesting to note in this connection that, of the 40 species here added to Smitt’s list, 33 are limited entirely or chiefly to a tropical distri- bution. The arrangement of the bryozoan species, genera, and families has under- gone so much alteration in recent years, as a result of the studies of Norman, Waters, Calvet, and Levinsen, as to be wholly unintelligible to anyone except the systematist familiar with the changes of the last ten years. It is im- possible that this should be otherwise, since the older classification was based almost entirely upon the structure of the skeleton of dead and dried material, without reference to the general morphology. In the following list I have followed as far as possible the classification adopted by Levinsen (1909). In the cases of a few imperfectly understood species the older genera have been retained as ‘“‘catch-alls.’’ Thus the genera (?) Lepralia and Phylactella have been used in the old Hincksian sense for certain species which, with more complete knowledge, must un- doubtedly go elsewhere. The figures were drawn by Mr. S. Shimitori, except figures 7, I1, 22, and 23, which are the work of Mr. H. Murayama. ENTOPROCTA. GENUS PEDICELLINA Sars, 1835. Pedicellina cernua (Pallas). OSBURN, 1912, p. 213, Synonymy and previous records of the occurrence of this species on the American coast. Apparently not common, but several small colonies were taken on the piles of docks and dredged at 10 fathoms. The Bryozoa of the Tortugas Islands, Florida. 185 GENUS BARENTSIA Hincks, 1880. Barentsia discreta (Busk). Busk, 1886, p. 44 (Ascopodaria discreta).—JULLIEN, 1888, p. 13 (Pedecellina austra- lis).—(?) VERRILL, 1900, p. 594 (Barentsia timida sp. nov.).—WATERs, 1904, p. 99.—OSBURN, I9QI2, p. 214. This species, first described from Tristan da Cunha in the Challenger reports, is now known to be widely distributed, as follows: Tristan da Cunha, 100 to 150 fathoms (Busk); China Sea, 27 fathoms (Kirkpatrick) ; Cape Horn, 26 fathoms (Jullien); Ile Londonderry, Magellanes, Chile (Waters) ; Woods Hole, Massachusetts (Osburn); Beaufort, North Carolina (Osburn); Tortugas, Florida (Osburn). If Verrill’s Barentsia timida from the Bermudas is the same species, as I strongly suspect, this will add another locality. Verrill states in regard to his B. timida that it is closely allied to B. discreta, but that the latter has a shorter and more annulated basal cylinder and also several annulations of the stem below the base of the cup. In my experience, the annulations of the stem are of little importance, if any, as there is much individual vari- ation which may represent only differences in contraction at this point. The individuals are small and the species may readily escape observation. It appears only once in my collection made at the Tortugas, being dredged in 18 fathoms directly north of the island on a bottom of coral mud; several fragments of colonies attached to shells. CYCLOSTOMATA. GENUS CRISIA (PART) Lamouroux, 1816. ? Crisia denticulata (Lamarck). LAMARCK, I816, p. 137 (Cellaria denticulata).—STIMPSON, 1853, p. 18.—SMITT, 1872, p. 4 (Crista eburnea).—JELLY, 1889, p. 73.—HARMER, I891, p. 136.—VERRILL, 1879, p. 28; 1900, p. 592.—WHITEAVES, I9QOI, p. I10.—OSBURN, I9I2, p. 216. Numerous small colonies were taken at from 10 to 15 fathoms in various places about the island, attached usually to sponges and shells. Smitt records the species as C. eburnea from 7 to 60 fathoms. Stimpson (1853), Verrill (1879), and Whiteaves (1901) have recorded it from New England to Canada, though these records are questionable on account of failure to consider the ocecia, and the writer has recorded it questionably from Woods Hole, Massachusetts (ocecia wanting). Verrill has reported it as common at Bermuda. Harmer (J. c.), referring to Smitt’s Florida record, states: ‘‘I do not feel certain that the form described is really identical with C. ramosa, although it can hardly be regarded as C. denticulata,” and suggests that it may be Stimpson’s C. cribraria. It does not seem to agree with the latter species, however, in any essential point (see Osburn, 1912, p. 215, for redescription of C. cribraria), nor does it agree with C. ramosa, in which the radical fibers have long internodes with yellow or colorless joints. It does agree with C. denticulata in having short internodes in the radical fibers and the 186 Papers from the Marine Biological Laboratory at Tortugas. joints of these fibers, as well as those of the zoarium, conspicuously jet- black. As the form of the colony and of the zocecia agree fairly with denticulata, I retain that name for the species. It is unfortunate that ovicells are wanting, so that it is impossible to make a positive determina- tion, for, in spite of the number of times the species has been recorded from North America, there seems to be not one of these records based on an unquestioned identification. GENus LICHENOPORA Defrance, 1823. Lichenopora hispida (Fleming). FLEMING, 1829, p. 530 (Discopora hispida).—JELLY, 1889, p. 134, synonymy. A very minute specimen was taken on an alga at 2 fathoms and placed with some question in this species. A well-developed colony taken by Dr. Paul Bartsch at Biscayne Key, Florida, has recently been examined, and after comparison there seems to be no doubt as to the identity of the Tortu- gas specimen. The species hitherto has not been noted in the Florida or West Indian regions. CHEILOSTOMATA. GENus AETEA Lamouroux, I8I2. Aetea truncata (Landsborough). LANDSBOROUGH, 1852, p. 288 (Anguinaria truncata).—JELLY, 1889, p. 5, synonymy and references.—CORNISH, 1907, p. 75.—NORMAN, 1909, p. 283. Common in shallow water and down to 5 fathoms, creeping over shells and seaweed. It has been recorded at Canso, Nova Scotia (Cornish, I. c.). Aetea sica (Couch). Coucu, 1844, p. 102 (Hippothoa sica).—JELLY, 1889, p. 5 (Atea recta) synonymy and references.—WATERS, 1909, p. 129 (A¢tea recta).—NORMAN, 1909, p. 283. Tortugas at 10 fathoms on shells. Not previously noted in American waters. Some authors prefer to consider this only a variety of tea anguina. The latter has not been taken in the Florida region, though it is common northward from Beaufort, North Carolina. GENUS BuGuLA Oken, I815. Bugula neritina (Linne). LinnE, 1758, p. 38 (Sertularia neritina).—VERRILL, 1878, p. 304 (Acamarchis neritina); 1900, p. 588.—ROBERTSON, 1905, p. 266.—WATERS, 1909, p. 135. This widely distributed warm-water species is abundant in the shallow water, growing on piles, shells, seaweed, etc. Colonies 1.5 inches in height with ocecia abundantly developed were taken on the bottom of a skiff which had been in the water only from May 1 to June 23. Verrill has recorded the species for the Bermuda Islands and from Fort Macon, near Beaufort, North Carolina. Otherwise it has not been noticed on the American side of the Atlantic. Robertson found it common on the Pacific coast as far north as Monterey Bay, California. The Bryozoa of the Tortugas Islands, Florida. 187 Bugula neritina var. minima Waters. WATERS, 1909, p. 136. One small colony with ovicells abundantly developed and with numerous avicularia (as described by Waters in Red Sea specimens) was taken at a depth of 8 fathoms. Previously known only from the type locality. This variety is distinguished from the typical neritina by the smaller size of its zocecia and by the presence of avicularia situated near the lower end of the zocecia. Bugula flabellata (Gray). Gray, 1847, p. 106 (Avicularia flabellata).—SMITT, 1872, p. 18.—VERRILL AND SMITT, 1873, p. 7II.—VERRILL, 1879), p. 189; 1880, p. 29 (Bugula flustroides).— OsBURN, I9I2, p. 225. A single small colony of this species, attached to Retepora ailantica, was taken at a depth of 12 fathoms. The ovicell is more widely opened than in Woods Hole specimens. A very widely distributed species, occurring northward on our coast to Maine. Miss Robertson found it at San Diego, California. Bugula microcecia n. sp. (Figs. | to 3.) pe Fic. 1.—Bugula microecia n. sp. Front view of portion of branch, showing details. Upper two ocecla not quite complete. Lower one complete with embryo. Fic. 2.—The same. Portion of main branch showing joints. Degenerate polypides are present. Fic. ar eae Portion of stem higher up, showing less-modified zocecia. Remains of degenerate poly- pides within. Zoarium delicate, composed of a central stalk with long, narrowly flabellate branches arising in an irregular dichotomous fashion from the main stem. The stalk, as well as its branches, consists of very much elongated and modified zocecia arranged biserially. That these are really zocecial in character is shown by the fact that they contain polypides and occasionally one of them even bears an avicularium. Notwithstanding this, definite joints appear at the bifurcations of the main and all the accessory branches. These conditions show an interesting connecting link 188 Papers from the Marine Biological Laboratory at Tortugas. between the Bugula and Stirparia types of stems. From the lower stem zocecia arise numerous, strong, radical fibers. The ordinary zocecia, which are arranged biserially, are long and slender and expand but little toward the tip; the frontal aperture extends almost to the base and faces somewhat toward the axis of the branch; a strong spine is situated at the distal outer edge; near the distal end, at the point where the outer wall curves farthest over the frontal area, is situated a short, stout avicularium with a strongly decurved beak; the avicularium is set upon a stalk so short as to be almost wanting. The ocecium is hemispherical, very small, and set very low down, par- tially hidden by the spine, so as to be very inconspicuous; it is thin and transparent, with a slightly arched aperture. Many of these contain em- bryos in various stages of development. Taken at 18 fathoms on a bottom of coral mud, several colonies attached to shell fragments; the largest 2.5 inches in height. Small fragments were also found decorating the legs of crabs (Hyas sp.?) at the same depth. Bugula caraibica Levinsen. LEVINSEN, 1909, p. 104, pl. 1m, figs. 2A—-2N. This species, recently described from St. Croix, Danish West Indies, is common at the Tortugas on the piles of the government docks, and colonies 1.5 inches high were taken on the bottom of a skiff which had been in the water only from May 1 to June 23, 1908. It is one of the most conspicuous species of the region, growing in loose tufts of a fine purple color to the height of 4 to 6 inches. Levinsen does not recognize Stirparia as a separate genus, but if the species with a jointed stalk are to be separated, caraibica will fall in that genus. Bugula armata Verrill. (Figs. 4 and 5.) VERRILL, 1900, p. 588 (Bugula [Caulibugula] armata n. sp.). Verrill has described from Bermuda a species which I take to be the same as one occurring at Tortugas. His description is as follows: A much more delicate, white Bugula, consisting of diverging, fan- like branches attached to the alter- nate sides and to the tip of slender jointed stems, sometimes having alternately a long joint and a very short joint, but more frequently the Fic. 4.—Bugula armata Verrill. Modified zocecia at base short joint is lacking and the end of ot Diane snosene arrangement of spines and position the joint is swollen, as in Stirparia. of avicularia. ‘ Fic. 5.—The same. At tip of same branch, showing character There are usually 2 or 3 annulations aS ae eat at the base of each main branch and these arise just below the internodes. Many of the cells have a slender, distal vibraculum or sometimes two. It should doubtless form the type of a new genus or subgenus (Cault- The Bryozoa of the Tortugas Islands, Florida. 189 bugula) intermediate between Bugula and Bicellaria on account of its articulated spines or vibraculum, and related to Stirparia by its jointed stem. It may be named Bugula (Cauli- bugula) armata. Its zocecia are oblong and biserial, alternate. The pedicellarie are on short pedicels, large, lateral not numerous. Verrill in the above description overlooked some interesting and im- portant points. He makes no mention of radical fibers which originate at the lower ends of the internodes of the stalks. He does not mention the ocecium, which is subglobose with a rather wide opening directed toward the zocecial axis. The ocecium is attached by a narrow stalk to the outer edge of the zocecial aperture near the distal extremity and is turned sidewise at a right angle to the zocecial axis. The avicularia (‘‘pedicellariez”’ of Verrill) show a peculiar distribution. The basal zocecium bears no avicularium, but the zocecia immediately above have the avicularia situated on the side about half-way along the aperture. The next few cells in order have the avicularium farther and farther removed toward the distal end, and in all zocecia farther out on the branches the avicularium is on the distal outer edge of the aperture. This is a feature which I have never seen exhibited by any bryozoa. There are one or two ong spines, jointed at the base, on the distal end of the zocecium (sometimes wanting). Verrill errs in calling these ‘“‘vibracula,’’ a term which should be applied only to modified avicularia with elongate mandibles. The basal zocecium of the main branches arising from the stem differs from the others in having usually 6 spines surrounding the aperture, 4 on the outer, 2 on the inner edge, a feature exhibited by Stirparia occidentalis (Robertson, 1905, pixie, fig:-73).. I can see no reason for erecting a new genus for this species. The jointed stalks, swollen at the nodes, seem sufficient to place it in Stirparia if that name were to be maintained for the Bugulas with jointed stalk. The jointed spines do not necessarily relate the species to Bicellaria, as this feature is present in some species of Bugula and Stirparia. Taken on a number of occasions at 8 to 10 fathoms, growing in a sprawl- ing fashion over sponges. Found also on the legs of spider crabs at Io to 18 fathoms, and dredged on a bottom of coral mud at a depth of 18 fathoms. GENus BEANIA Johnston, 1838. Beania mirabilis Johnston. JOHNSTON, 1847, p. 372.—JELLY, 1889, p. 17, for further reference.—ROBERTSON, 1905, p. 276. Several fragments of colonies on the legs of a crab (Hyas sp.) taken at 18 fathoms. This striking and well-known species has not before been noticed on the American side of the Atlantic. Beania intermedia Hincks. Hincxs, 18814, p. 133 (Diachoris intermedia).—WATERS, 1906, p. 15; 1909, p. 137+ At 5, 10, and 15 fathoms, sprawling over hydroids and other bryozoa, and on shells. Very small colonies consisting of only a few cells. The 190 Papers from the Marine Biological Laboratory at Tortugas. simple, spineless character of the zocecium easily serves to distinguish it from others of the genus. The species is entirely southern in its distribution, being recorded from Tasmania (type locality), Australia, Chatham Island, Ganjam coast, and the Red Sea. The present record therefore adds the species to the Atlantic fauna and greatly increases the known range. Hincks described the species as having spines on each side. Waters (1909) remarks that they are absent in Red Sea specimens, but sometimes present in those from Chatham Island. There is no indication of them in my material. Beania cupulariensis n. sp. (Figs. 6 and 7.) Small, straggling colonies growing attached to the under side of Cupularia guiniensis. ‘The individual cells are fairly large (about as in B. mirabilis). On each side of the flattened frontal area there are about 6 (5 to 7) slender marginal spines which curve somewhat over the area. At the distal end 2 smaller spines project forward. At each side of the aperture is placed an ( ee S 6 Sr 7 oe 6 Fic. 6.—Beania cupulariensis n. sp. Mode of branching and details of zocecium, SS | Fic. 7.—The same. Dorsal side, showing zocecial base and point of origin of W/, ae dorsal fiber. avicularium situated on a short stalk on margin of cell; as far as observed, these are always paired. The form is short, the beak strongly decurved. The zocecium arises from the preceding one of the series, near the distal part of the dorsal surface, by a tubular stalk so short that the top of one zocecium projects over the base of the next in series. A branch arises about midway of the zocecium near the lateral margin. Here again the tubular portion is very short and, as far as observed, only one branch arises from a zocecium—never in pairs. There seem to be no other processes arising from the zocecium and many of the cells have no tubes, branches, or other processes arising from them, except that of the zocecium next in the series and a dorsal process for attachment. The Bryozoa of the Tortugas Islands, Florida. I9I The colony appears to be very loosely attached to the Cupularia by processes from the dorsal sides of the cells. The species occurred at various depths—in fact, wherever this species of Cupularia was found, at from 10 to 22 fathoms, but was found nowhere else. Usually only a single colony was found on one Cupularia, but occasionally on a large specimen two or three colonies would occur. GENUS SYNNOTUM Hincks, 1886. Synnotum aviculare (Pieper). PIEPER, 1881, p. 43 (Gemmellaria avicularis).—HINCKS, 1886, p. 257-—ROBERTSON, 1905, p. 286. This well-known species is abundant about the islands, growing espe- cially on shells and sponges at a depth of 8 to 10 fathoms. It has not before been recorded from North America, except on the Pacific coast, where Miss Robertson found it at San Pedro and San Diego, Southern California. Hincks (1886) described the sessile avicularia as alternating on Adriatic specimens. Robertson states that they appear on every pair in specimens from California. In the Florida material in my collection the alternate condition is the rule, but occasionally sessile avicularia appear on successive pairs of zocecia. Besides the above localities the species is now known from the Mediterranean and Red Seas, South Africa, Australia, and the Andaman Islands. Genus NELLIA Busk, 1852. Nellia oculata Busk. BuSK, 1852, p. 18.—LAMARCK, 1816, p. 135 (Cellaria tenella).—SmirT, 1873, p. 3.— JELLY, 1889, p. 94 (Farcimia tenella, references and synonymy).—WATERS, 1909, p. 167 (Farcimia oculata).—LEVINSEN, 1909, p. 120 (Nellia tenella). Abundant at 10 to 18 fathoms, frequently attached to sponges or occasionally to shells. The usual height of the colonies is 0.5 to 1 inch. One large, branched colony was taken, measuring 4 inches across and of equal height. Smitt recorded the species from 13 to 138 fathoms. It has also been recorded from Texas and St. Thomas, West Indies (Levinsen), and is widely distributed in the Atlantic, Pacific, and Indian Oceans down to a depth of 550 fathoms. Waters (1909, p. 167) objects to the use of Lamarck’s name ¢enella on the ground that the short description given by Lamarck is not diagnostic and ‘‘might be applied to several species or even genera.” GENUS SCRUPOCELLARIA van Beneden, 1844. Scrupocellaria cornigera (Pourtales). POURTALES, 1867, p. 111 (Canda cornigera).—SmITT, 1872, p. 14 (Cellularia cornigera).— JELLY, 1889, p. 61 (Cellularia cornigera). One small specimen of this species was taken at 10 fathoms and several others at 15 fathoms. The radical fibers are characteristically armed with stout, retrorse spines, as figured by Smitt. This alone is not sufficient for determining the species, as Busk has noticed serrate radicals in S. ferox 192 Papers from the Marine Biological Laboratory at Tortugas. Busk and S. macandrei Busk, and Waters has named a species from the Red Sea S. serrata on account of this character. Florida specimens of S. cervicornis Busk show similar spines occasionally on the basal radicals. S. cornigera is a very delicate species and the radical fibers are transparent and thread-like, and this fact alone will serve to distinguish it from S. cervicornis. Pourtales took the species at 270 fathoms, so it has a rather wide range in depth and temperature. Scrupocellaria cervicornis Busk. Busk, 1852, p. 24.—SMITT, 1872, p. 14 (Cellularia cervicornis).—VERRILL, 1900, p. 594. Occurs commonly about the islands, from low water to 18 fathoms, growing on piles of docks, attached to stems of gorgonias, etc. Colonies about 0.5 inch in height, with ocecia containing eggs, were taken from the bottom of a skiff on June 23, which had been in the water only since May 1. Recorded by Smitt from 7 to 17 fathoms. The radical fibers, which are abundant at the base of the colony, are usually smooth, but in a few cases they bear retrorse spines like those of S. cornigera. In such cases the species are easily distinguished by the fibers alone, as they are much coarser in cervicornis. The cervicorn spines on the outer distal extremity of the zocecia are well marked. GENus CANDA Lamouroux, I816. Canda caraibica Levinsen. LEVINSEN, 1909, p. 142. This species has been recently described by Levinsen, who gives no further indication of its distribution than is embodied in his remark “ plenti- ful material of a West Indian species.” Levinsen suggests the probability that this species is identical with C. simplex Busk, but being unable to determine this point he gives a new name to the West Indian material. Several colonies were taken by the writer at the Tortugas at a depth of 15 fathoms, attached to shells. The largest of these was not much over 0.25 inch in height and no ovicells nor avicularia were present. The absence of an opercular spine and the length of the membranous frontal area (two-thirds of the whole zocecium) show this form to be that described by Levinsen. The vibraculum in some cases is longer than the width of the branch. The other Florida species of this genus, C. retiformis Pourtales (1867, p. 110), which Levinsen wrongly attributes to Smitt, is quite distinct from C. caratibica, since it possesses a broad opercular spine, the membranous area is but little more than half as long as the zocecium, and there are differences in the vibracula. C. retiformis was not found at the Tortugas. The Bryozoa of the Tortugas Islands, Florida. 193 GENUS MEMBRANIPORA Blainville, 1834. Membranipora membranacea (Linné). LinnE£, 1766-68, p. 1301 (Flustra membranacea).—PACKARD, 1867, p. 274.—ROBERTSON, 1908, p. 267.—NORMAN, 1909, p. 286. One colony on the bottom of a skiff. In every respect this colony agrees with European specimens in my collection, except that there is a very slight extension of the calcification over the area at the base of some of the zocecia. This is so slight, however, that I can not consider it of specific value. The species was not listed for Floridian waters by Smitt, nor was it taken by the writer at Woods Hole, Massachusetts, in several years collecting. Packard recorded it from the Labrador coast. It isa common European species, occurring as far south as Madeira (Norman). It is also found in Australia and New Zealand, and Miss Robertson (1908) states that on the Pacific coast of North America it occurs from California to Alaska. ? Membranipora lacroixii (Audouin). AUDOUIN, 1826, p. 240 (Flustra lacroixii).—BUSK, 1852-4, vol. 2, pl. 1—DAwson, 1859, p. 256.—PACKARD, 1867, p. 8.—SMITT, 1873, p. 18.—JELLY, 1889, p. 162 (M. reticulum).—WATERS, 1898, p. 697.—WHITEAVES, 1901, p. 97.—ROBERT- SON, 1908, p. 261.—OSBURN, IQI2, p. 227. Taken in drift incrusting the carapace and legs of a large crab, and at 8 fathoms on shell. The specimens taken at Tortugas are much more heavily calcified than those from Massachusetts and between the zocecia are numerous, roughly triangular prominences with triangular, membranous areas, as figured by Busk (l.c., pl. 1), while these are almost wanting in the Massachusetts specimens. Examination of the reverse side of the zocecium, however, reveals the transparent circles and the irregular projections described by Waters (I. c.). There has been much difference of opinion concerning this species and there can be but little certainty in regard to many earlier records, such, for example, as those of Dawson and Packard. Smitt’s figures of Florida specimens seem, without question, to represent the same species as my own, though the dorsal surface is not shown. The range on the eastern coast of North America is from Florida to the Gulf of St. Lawrence, or to Labrador, if Packard’s record is valid. Elsewhere it is known from Europe, New Zealand, and from California northward to Alaska. Membranipora tehuelcha (d’Orbigny). D’ORBIGNY, 1839, pl. 17 (Flustra tehuelcha).—Bosc, 1802, p. 118 (Flustra tuberculata). —JELLY, 1889, p. 167 (M. tehuelcha) and p. 168 (M. tuberculata).—WATERS, 1898, p. 674.—VERRILL, 1900, p. 594 (Bzflustra dentata).—NORMAN, 1909, p. 286 (Membranipora tuberculata).—OSsBURN, 1912, Pp. 231. This species is found on Sargassum bacciferum (gulfweed) wherever it floats. It is abundant at the Tortugas. Verrill notes its occurrence at Bermuda, under the name of Biflustra dentata, as ‘‘common on Sargassum found upon the beaches.’’ Smitt did not record it for the Florida region. 14 194 Papers from the Marine Biological Laboratory at Tortugas. Norman prefers to use Bosc’s older name for this species on the ground that, while his description means nothing, Bosc stated that it occurs ‘“‘en immense quantité sur les fucus nageans dans |’Atlantique.’’ As I am not aware that, under the rules of nomenclature, the habitat alone is considered a sufficient diagnosis, I prefer to follow Waters in using d’Orbigny’s name. Membranipora irregularis d’Orbigny. D’ORBIGNY, 1839, p. 17, pl. vIn, figs. 5 and 6.—Smi1T, 1873, p. 8, pl. 11, fig. 63.— JELLY, 1889, p. 151, for synonymy.—WATERS, 1904, p. 31. Incrusting shells at 8, 15, 18, and 22 fathoms. Smitt’s figure is very satisfactory and there can be no doubt that the present species is the one which he figured as taken at 60 fathoms. The granulation of the border varies with the amount of calcification. Ocecia are present in some numbers. In younger stages these are quite prominent, but with later calcification they become included in the general crust. Membranipora savartii (Audouin). AUDOUIN, 1826, p. 240 (Flustra savartit)—Smitt, 1873, p. 20 (Biflustra savartii).— WATERS, 1909, p. 137. One small, branching specimen 0.5 inch in height was collected at a depth of 10 fathoms. The broad, internal, basal denticle figured by Waters for Red Sea specimens is the best means of determining the species. Recorded by Smitt, 29 fathoms. GENUS CUPULARIA Lamouroux, 1821. Cupularia guiniensis Busk. Busk, 1854, p. 98.—Busk, 1859, p. 87 (C. canariensis).—SMITT, 1873, p. 10 (Mem- branipora canariensis).—NORMAN, 1909, p. 289, synonymy. Abundant at 10 fathoms, especially on soft bottom with alge. The umbrella-shaped disks range in size from 0.125 to 0.75 inch in diameter. The color of living colonies is horn brown, due mainly to the chitinous bristles which form the mandibles of the avicularia. When touched the bristles stand erect for some time. Recorded by Smitt from Pourtales’s collections from 10 to 44 fathoms. Smitt listed this species under Membranipora because of the character of the zocecium, holding the zoarial characters as of no importance. Just why he should have retained the related C. lowei and C. doma in Cupularia is not certain, but probably on account of the greater amount of calci- fication of the front wall. The under surface of the colony is usually clear and free of any attached animals or plants, except that in numerous cases there occurs, closely attached, a new species of Beania, which I have named B. cupulariensis. Cupularia lowei Busk. Busk, 1854, p. 99.—SMITT, 1873, p. 14 (C. umbellata Manzoni).—VERRILL, 1878, p. 305. (C. umbellata)—NoORMAN, 1909, p. 290. Only two specimens, both dead, were taken, one at 12 and the other at 22 fathoms, in company with C. guiniensis. Each was about 0.25 inch in. diameter. The Bryozoa of the Tortugas Islands, Florida. 195 Recorded by Smitt for 29 fathoms and also in 7 fathoms off the mouth of the Cape Fear River. Smitt regarded C. johnsoni Busk (syn. C. doma d’Orbigny) as a highly calcified form of C. lowei (umbellata), but later inves- tigations do not support this view. Verrill (1. c.) recorded the species with doubt from one worn specimen taken at Fort Macon, near Beaufort, North Carolina. As the writer has taken specimens at the same locality, Verrill’s record may be considered good. GENUS CRIBRILINA Gray, 1848. Cribrilina floridana (Smitt). SMITT, 1873, p. 23 (Cribrilina figularis and local variety floridana).—JELLY, 1879, p. 66 (calls attention to fact that C. figularis of Smitt is not Johnston’s species, and lists C. floridana as a distinct species). A number of small colonies of this species taken at 5 to 15 fathoms on shells. Smitt recorded it from 29 to 42 fathoms. It is not known to occur outside of the Florida region. No ocecia nor avicularia are developed on my specimens. The posterior margin of the orifice is straight, and the orifice is completely filled by the semicircular operculum, which is well chitinized and of a brownish color. There are two short, stout oral spines and in young zocecia there is fre- quently a third median spine. There is a possibility that this species may belong to the genus Puellina of Jullien as amended by Levinsen (1909, p. 159), but the immature colonies in my collection seem rather to belong to Cribrilina in the strict sense. GENus ARACHNOPUSIA Jullien, 1888. Arachnopusia monoceros (Busk). Busk, 1854, p. 72 (Lepralia monoceros).—JELLY, 1889, p. 67, synonymy.—LEVINSEN, 1909, p. 160 (discusses relationship and adopts Jullien’s genus Arachnopusia with emendations). A single specimen of this widely distributed species was taken at 5. fathoms, incrusting a shell. The colony was small, measuring only 0.125. inch across, but the zocecia ranged in development all the way from the ancestrula to adults with ovicells and from these to marginal individuals in various conditions of calcification. The species has not before been taken in North American waters. GENUS SMITTIPORA Jullien, 1881. Smittipora abyssicola (Smitt). SmittT, 1873, p. 6 (Vincularia abyssicola).—JELLY, 1889, p. 253, synonymy and refer- ences.—HINCKS, 1881, p. 155; 1887, pp. 161 and 164. Found at low water and down to 15 fathoms on shells, bases of gorgonias, etc. Smitt recorded two specimens taken by Pourtales, one at 68 fathoms, the other at 450 fathoms. The bathymetrical range of the species, from low water down to nearly 0.5 mile, is worthy of note. Only the incrusting phase was seen, with no evidence of the erect stems described and figured by Smitt in one colony. The family relations of this 196 Papers from the Marine Biological Laboratory at Tortugas. genus are very uncertain. Hincks (1881, p. 155) placed it in the Micro- poride, but later (1887, p. 154) he removed it from this family on account of ‘‘the double layer of the ectocyst,’’ and placed itin the Steganoporellide. As we understand the latter family at present, this genus can not be included init. It will require a careful study of sectioned material to determine the real position of the genus. GENUS STEGANOPORELLA Smitt, 1873. Steganoporella magnilabris (Busk). Busk, 1854, p. 62 (Membranipora magnilabris).—SmITT, 1873, p. 15 (Steginoporella elegans).—VERRILL, 1900, p. 594 (Steginoporella elegans)—HARMER, 1900, pp. 279-286 (revision, synonymy, and detailed description) —LEVINSEN, 1909, pp. 167-168 (discussion of family and genus). Common in low water to 15 fathoms. Beautifully convoluted, frill-like colonies were taken on piles and on shells, sponges, coral, etc., at 12 to 15 fathoms. The erect frill-like extensions are sometimes 2 inches high and 3 inches broad. These are bilaminate and are sometimes tubular in form. The color in life varies from pink to reddish brown. Smitt recorded the species in Pourtales’s collections from 15 to 37 fathoms; Bermuda (Verrill) ; other distribution given by Harmer (I. c., p. 280). Steganoporella connexa Harmer. HARMER, 1900, p. 254. Dead colonies of what appears to be this species were taken incrusting shells at a depth of 12 fathoms. The opercula are all lost from the colony, so it is impossible to make use of this important structure for identification. The character of the calcification, with the junction of the median process with the sides of the zocecium to form a pair of opesiule or foramina, is similar to that described and figured by Harmer. Since this connection of the median process with the zocecial walls is known only in S. connexa, I refer my specimens to that species. The other points, as far as I can make them out on the dead specimens, agree with connexa, except perhaps in the matter of size, since the zocecia in my specimens are smaller than described by Harmer. There is much variation in the matter of size, however, a condition also stated by Harmer to occur in connexa. Only one form of zocecium can be distinguished. The species is known from a single locality, ‘‘John Adams Bank,” the location of which is somewhat in doubt, but Harmer seems to feel satisfied that it is south of Abrolhos Island off the coast of Brazil. GENUS THALAMOPORELLA Hincks, 1887. Thalamoporella rozierii (Audouin). AUDOUIN, 1826, p. 239 (Flustra rozierit).—(?) SMITT, 1873, p. 16 (Steginoporella rozi- erit).—LEVINSEN, 1909, p. 181 (discusses at length, with five varieties). Found on shell in drift and at 10 fathoms on sponges. Only small, unilaminar colonies without ocecia were taken. Smitt found only a single, small specimen in Pourtales’s collections. His description of this specimen The Bryozoa of the Tortugas Islands, Florida. 197 is brief and his figure (pl. 4, fig. 102) does not show the avicularium nor the spicules, so I include it in the above synonymy with some doubt. I have received from Professor A. E. Verrill a fine, erect, convoluted, bilaminate colony taken on the west coast of Florida. Levinsen (I. c., p. 183) records the variety labiata from Jamaica and my Florida specimens agree fairly closely with this variety. Thalamoporella granulata Levinsen. LEVINSEN, 1909, p. 188. In the drift were found several dead specimens incrusting shells and calcareous alge. The sinuated character of the ori- fice with the strong lateral denticles; the nearly equal opesiule reaching the mar- 8 ginal wall and with granulated edges; the dorsal opesiular outgrowths; the P'qyo Taner sores Gomuaa Levinsen: granulated character-of the zocécial walls; Ce2qbet 1s closed basally," except for'two the form of the large, spatulate avicularian mandible, and the character of the spicules, all approach closely to T. granulata. The only difference to be observed is in the basal portion of the avicularian chamber. When completely calcified this structure becomes closed by a calcareous lamina up to the hinge denticles of the mandible, leaving only two openings similar in appearance to the opesiulz of the ordinary zocecia, except that they are much smaller and are removed from the lateral wall. Thalamoporella falcifera (Hincks). HINCKS, 18808, p. 380 (Steganoporella rozieri, form falcifera).—LEVINSEN, 1909, p. 186. A single specimen of this species, which is distinguished by the falcate mandible of the avicularium, was taken in shallow water growing on alge. Apparently this species grows only on alge, if one may judge from the comparatively small number of records. It has a wide distribution and is recorded from Campeche Bank, Yucatan; Java Sea; Geographe Bay, Aus- tralia, and from latitude 23° 30’ N., longitude 40° W. GENUS SAVIGNYELLA Levinsen, I909. Savignyella lafontii (Audouin). AUDOUIN, 1826, p. 242 (Eucratea lafontii).—JELLY, 1889, p. 93 (synonymy under Eucratea lafontit) NORMAN, 1909, p. 295 (Catenaria lafontit, later synonymy and discussion of relationships)—WATERS, 1909, p. 131 (Cafenaria lafontiz, later synonymy and discussion of generic name).—LEVINSEN, 1909, pp. 213 (note) and 274 (Savignyella, new genus and reasons for change in generic name). This widely distributed tropical species is abundant about the islands from low water down to 10 fathoms on shells, piles, and sponges, in company with Amathia, Bowerbankia, etc. Ocecia with eggs in various stages of 198 Papers from the Marine Biological Laboratory at Tortugas. development are common. A number of colonies with ocecia containing eggs were on June 23 taken from the bottom of a skiff which had been in the water only since May 1. The color in life is dark brick-red, which makes the colonies quite conspicuous against the background in spite of their small size. The presence of this species in American waters has hitherto escaped notice. While there has never been any question as to the validity of this spe- cies, there has been great confusion as to its generic position ever since its discovery. This has been sufficiently discussed by Norman (I. c.), Waters (I. c.), and Levinsen (I. c.). Waters remarks ‘‘though it certainly never ought to have been placed under Catenaria, it seems best to leave it here,” but Levinsen solves the problem by the erection of a new genus Savignyella. This seems the only proper course, since the species does not have generic relationship with any of the older genera (Catenaria, Eucratea, Alysidium, Catenicella) in which it has been placed by previous authors. GENuS HiprpoTHOA Lamouroux, 1821. Hippothoa distans MacGillivray. MACcGILLIvRAY, 1868, p. 130.—JELLY, 1889, p. 112 (Hippothoa flagellum), references.— WATERS, 1904, p. 54, synonymy and references. This minute species occurs abundantly at the Tortugas from low water to 12 fathoms, spreading over the inside of dead shells. Ocecia are abun- dantly developed. The ‘‘short processes arising from the side of the zocecium, below the lateral tubular branches,” described and figured by Waters (l. c.) as occurring on specimens from Sydney, New South Wales, are developed on many of the zocecia. JI am unable to add anything to Waters’s descrip- tion of this organ, except that it is constricted at the base where it is attached by a movable joint to the zocecium. The species is very widely distributed in European seas, New Zealand, Australia, Pacific Ocean, Cape Horn, etc., but has not hitherto been noticed on the American side of the Atlantic. GENUS TRYPOSTEGA Levinsen, 1909. Trypostega venusta (Norman). NORMAN, 1864, p. 84 (Lepralia venusta) and 1909, p. 299.—GABB and Horn, 1862, p. 127 (Lepralia tnornata).—SmitT, 1873, p. 37 (Gemellipora glabra, forma striatula) and p. 61 (Lepralia inornata Gabb and Horn).—JELLy, 1889, p. 233 (Schizoporella striatula), p. 237 (S. venusta) and p. 128 (Lepralia inor- nata).—LEVINSEN, 1902, p. 23, and 1909, p. 281 (Trypostega venusta). On shells, Steganoporella and Oculina, at 5 to 15 fathoms. Smitt recorded it as Gemellipora glabra forma striatula, incrusting nullipores at a depth of 68 fathoms, one colony, and as Lepralia inornata, two colonies from 26 and 60 fathoms. After careful study of the material at hand, I am unable to distinguish more than one species, or to separate it from the venusta of Norman, even as a variety. The form of the aperture, the umbonate knob below, the puncturing, the character of the dwarf zocecia and their relation The Bryozoa of the Tortugas Islands, Florida. 199 to the ocecia, all appear to me to be identical. Levinsen (see above) has recently made this species the type of a new genus, Trypostega, characterized especially by having the ocecia covered by dwarf zocecia with scattered pores and a minute orifice. This species is widely distributed, but on the American coast is known only from the Florida region. Levinsen (p. 281) included Gabb and Horn’s and Smitt’s records for Lepralia inornata, but failed to include Smitt’s Gemellipora glabra forma striatula as a synonym. Smitt (pp. 61-62) sepa- rated the species on the following ground: ‘‘The most characteristic differ- ence, which has caused us to place them in different families, depends on the shape of the zocecial aperture,” and his figures show some differences, but these may be accounted for by variation. Ina single colony in my posses- sion the variation is as great as that shown by Smitt’s two specimens. GENUS ADEONA Busk, 1884. Adeona violacea (Johnston). JOHNSTON, 1847, p. 325 (Lepralia violacea).—(?) REuss, 1847, p. 85 (Cellepora heckeli).— SMITT, 1873, p. 30 (Porina violacea and Porina plagiopora Busk).—JELLY, 1889, p. 184 (Microporella heckeli), synonymy.—VERRILL, 1901, p. 54 (Porina plagiopora).—NORMAN, 1909, p. 296 (Reptadeonella violacea).—LEVINSEN, 1909, p. 283, discusses genus and species. Taken from 5 to 18 fathoms. Both the typical form and the nominal variety plagiopora (Busk) occur and in some colonies the avicularia are intermediate in position between that of violacea and plagiopora. Occa- sionally the characters of both “‘varieties,’”’ together with intermediate con- ditions, may be observed in the same colony. The usual color is purple, varying to nearly black, or through pale bluish to white. Smitt recorded the species ‘‘from the depth of 35 fathoms W. off Tortu- gas’’ (Porina violacea), and ‘‘at a depth of 60 fathoms W. off Tortugas” (Porina plagiopora), and Verrill has noted the form plagiopora at Bermuda. Otherwise the species has not been recorded from American waters. Levinsen (1909, p. 282) has returned to the use of Busk’s family Ade- onide, thus again breaking up the arrangement established by Hincks in his family Microporellide. The latter family was based on the presence of a frontal pore (ascopore) as the principal character; but, as Levinsen has shown, this brought together a number of forms which are separated by more fundamental characters. GENUS BRACEBRIDGIA MacGillivary, 1886. Bracebridgia subsulcata (Smitt). SmITT, 1873, p. 28 (Porina subsulcata).—VERRILL, 1901, p. 54 (Porina subsulcata).— HInNcKS, 1880a, p. 76 (Microporella subsulcata). Abundant from 10 to 12 fathoms. Branching colonies reach a height of 2inches or more. The color in life varies from yellowish pink to orange; when dead, white. Smitt recorded the species from 10 to 48 fathoms and a dead specimen from 471 fathoms. Verrill has recorded it from Bermuda, but otherwise it is known only from the Florida region. 200 Papers from the Marine Biological Laboratory at Tortugas. I have placed this species in the genus Bracebridgia with some hesitation, since, as this genus is understood, it possesses no ascopore (Levinsen, 1909, pp. 283, 289). In subsulcata a minute pore is usually present below the suboral avicularium; a similar pore appears above the basal avicularium when this organ is present. This latter pore was not figured nor mentioned by Smitt. I have not been able to determine positively the nature of this structure and it may be, from the fact that it is often wanting and also because of its presence in connection with the basal avicularium, that it is not an ascopore at all, or that it represents this organ in a vestigial condition. The nature of the colony and the zocecia with the marginal and other ribs, the form of the orifice, the presence of a line of special independent avicularia on each edge of a branch, all seem to agree with Bracebridgia. B. pyriformis (Busk) has a flattened area below the orifice where the avicu- larium is located in subsulcata, but the fact that MacGillivray found a single avicularium in this position in one colony of B. pyriformistis sufficient to show the relationship. Moreover, in the rare cases in which the oral avicularium is wanting in subsulcata there is a flattened area similar to that of pyriformis. GENUS RETEPORA Smitt, 1867. Retepora marsupiata Smitt. SMITT, 1873, p. 67.—Busk, 1884, p. 116 (Relepora atlantica)—GABB AND Horn, 1862, p. 138 (Phidolophora labiata).—JELLY, 1889, p. 212 (Jelly adopts Busk's. name délantica for this species, but Smitt’s publication of the name marsupiata clearly has priority). Taken on a number of occasions at 10 to 18 fathoms. The largest colony measured about an inch in height. The color in life is a delicate pink. Smitt records the bathymetrical distribution as 16 to 262 fathoms. GENUS RHYNCHOZOON Hincks, 1891. Rhynchozoon tuberculatum n. sp. (Fig. 9.) Zocecia small, rather evenly swollen, separated by slightly raised marginal walls; the surface, ex- cept in young individuals, strongly tuberculate. Pores are wanting, except for occasional very minute ones at the margin. Orifice ovate, the broader distal border often somewhat straight; at one side near the proximal border a strong pointed tooth extends often more than half-way across the orifice and curves backward; on the op- posite side a minute projection sometimes appears, fy. 9.—Rhynchosoon _ tubercula- but is often entirely absent. The peristome is thin (ft, G0. nowins Characters and raised high above the primary orifice, usually "4 ovicell- erect at the sides, but flaring slightly backward behind the orifice; above the large tooth the peristome appears to bear a minute, short-elliptical avicularium, but I am not able to distinguish a mandible in my specimen. The Bryozoa of the Tortugas Islands, Florida. 201 The ovicell is globose, at first smooth, but later tuberculate like the zocecium; on either side at the widest part and near the base is a rounded thin area appearing membranous; in complete calcification the peristome rises over the front of the ovicell and fuses with the outer layer, leaving a thin, somewhat triangular area on the front. One colony at 18 fathoms, incrusting a shell with a single layer of zocecia. Rhynchozoon solidum n. sp. (Figs. 10, 11, and 12.) pte: ( 10. Fic. 10.—Rhynchozoon solidum n. sp. Enlarged, showing primary aperture, spines, two pairs of denticles, and the beaded vestibular arch. Fic. 11.—The same. Operculum. i rn 5 Fic. 12.—The same. Fully calcified, showing avicularium, peristome, and ovicell. Zocecia small, little swollen, rather broad, closely set in a continuous crust, thick-walled, the surface smooth or irregularly traversed by very fine lines. As a rule only three pores are seen, one at the posterior margin and the others on either side of the orifice at the margin. Orifice evenly rounded, a little broader than long, with a rounded sinus in the proximal border. This sinus is really double, being formed by two sets of denticles, the lower pair equal in size and inclosing a semicircular sinus much like that of many Schizoporellas. Immediately above them is another pair of denticles, more pointed and a little longer than the lower ones, and often one of them is a trifle larger than the other; they are turned inward more than the lower pair, so as to inclose a narrower and more nearly circular sinus. The operculum is well chitinized, yellowish in color, the dots showing attachment of muscles situated nearly half-way from the circumference toward the center. Through the operculum can be seen a beaded vestibular arch very similar to that figured by Levinsen (1909, pl. XXIII, fig. 4e) for R. angulatum. In young cells there are four short, stout spines about the anterior margin of the orifice. The peristome is high and thick, extending upward and forward on the sides into thick, lappet-like processes, usually a little unsymmetrical; posteriorly the peristome extends upward and over the orifice in the form of a broad mucro, though sometimes this is but little developed. Small avicularia, sometimes a pair, frequently only one, are placed posteriorly not far from the margin, with the triangular beak pointing laterally. 202 Papers from the Marine Biological Laboratory at Tortugas. The ocecia are globose, raised, smooth, without pores; a thin extension of the peristome runs upon the top and fuses with the outer layer of the ovicell, leaving a thinner area upon the front. One small colony incrusting a shell at 8 fathoms. ARBORELLA, NEW GENUS. Zoarium erect, dichotomously branched, with flexible corneous joints at the bifurcations. Zocecia arranged in pairs, back to back, each pair facing at right angles to the preceding pair of the series, giving a somewhat quadrangular appearance. Front wall porous, but without special en- larged pores. Fertile zocecia shorter than the infertile. Orifice with a distinct sinus. Ovicell external, but inserted partly under the front of the zocecium, its orifice closed by the zocecial operculum. Spines and avicu- laria absent. A clear, chitinous, uncalcified ectocyst covers the zocecium and ovicell. The relationships of this genus can not as yet be stated positively. The general appearance of the colony and especially the character of the joints and the membranous ectocyst seem to relate it to Tubucellaria, but the character of the ocecium and the manner of origin of the zocecia at the tips of the branches, together with the absence of a special median frontal pore, distinctly separate it from this genus. It may be necessary to erect a new family to accommodate it. Sectioned material will be studied to determine the points necessary for a more complete understanding of the relationships. Arborella dichotoma n. sp. (Figs. 13 to 15.) Zoarium erect, dichotomously branching, with hinged, flexible joints at the points of bifurcation; internodes consisting of 8 to 18 pairs of zocecia, each pair set slightly in advance and at right angles to the preceding pair of the series, facing in four directions. Zocecia broad fusiform, wedge- shaped at the base where they are inserted between the preceding pair. The front wall evenly arched, cryptocyst well calcified, porous, with no specialized, larger pores. Ectocyst uncalcified and covering the calcareous wall as a clear layer. The orifice is evenly rounded in front and on the sides, somewhat straighter on the posterior margin, with a broad, shallow, but very evident sinus. The operculum is compound, well chitinized, with an arched rib on either side, extending inward and upward from the hinge denticle. Avicularia and spines are wanting. The fertile zocecia are some- what different in form from the ordinary ones, being shorter and somewhat broader, and the ocecium incloses nearly all of the orifice. The ocecium is rounded in outline, large and prominent, the aperture looking upward and closed by the zocecial operculum. The ocecial wall is porous like that of the zocecium and is covered by the clear ectocyst. The colony was evidently broken off in dredging and no description can be given of the base or the mode of attachment. The Bryozoa of the Tortugas Islands, Florida. 203 One colony 0.5 inch high, composed of 24 internodes, taken at 10 fathoms on a sponge bed between Loggerhead and Garden Keys, Tortugas. Fic. 13.—Arborella dichotoma n. sp. Portion of colony, showing mode of growth. Fic. 14.—The same, showing details. Fic. 15.—The same, operculum. GENUS TUBUCELLARIA d’Orbigny, 1850-52. Tubucellaria cereoides (Solander). SOLANDER, 1786, p. 26 (Cellaria cereoides).—WATERS, 1907, p. 129.—LEVINSEN, 1909, P- 305- One small colony about an inch in height was taken at a depth of 15 fathoms. This species has not before been taken on the American side of the Atlantic. Waters makes T. opuntioides Pallas a synonym of cereoides, but Levinsen believes that they should be kept separate. This difference of opinion, which has existed since the discovery of the form, does not seem likely to reach a settlement soon. The single specimen taken by me at the Tortugas is not sufficient to decide the point. It agrees closely with specimens of cereoides of the same size taken at Naples, but cereoides has hitherto been limited to the Mediterranean by those who separate the species, while Atlantic specimens are considered to be opuntioides. GENUS ESCHARELLA (PART) Gray, 1848. Escharella costifera n. sp. (Fig. 16.) Zocecia of moderate size, rather regularly ovoid in form, evenly swollen, nearly hyaline. Around the margin is a row of rather large pores, between 204 Papers from the Marine Biological Laboratory at Tortugas. which rise strong, conspicuous ribs which converge regularly toward the central portion of the front wall. They fade out, however, before they reach it, leaving a smooth central area devoid of ribs. The orifice, which is placed far forward, is rounded in front, more straight on the posterior border, and the hinge denticles are rather conspicuous. A few zocecia seem to show a slight median denticle, but I can not be certain of this. The peristome, which is very thin, rises in front and on the sides into a short tube, and is beset with six or eight long spines which are jointed at inter- vals. Posteriorly the peristome rises into a high, pointed mucro which projects forward over the ori- fice. This mucro is flat and thin, with a strong rib on each edge, looking as though it originated from two spines fusing at the tip, leaving an open space between. The development of the mucro, however, Fic. 16.—Escharella costifera ote : ; . sp. Details of front shows that this is not the case. The thin area forming wall, mucro, avicularium, i : geell sacere ova the middle of the mucro is rounded below and pointed arch (ia upper figure), above and looks as though it were meant to harbor ie or ee an avicularium, but no such structure is present. Avicularia are present on the zocecium, usually situated near the margin on either side of the mucronate process. Occasionally only one is pres- ent. The avicularium is comparatively large, the mandible long triangular, projecting sidewise or a trifle backward and pointing strongly upward, The ocecium is rounded in outline, prominent, imperforate, smooth on the upper surface, or with fine, radiating lines. Around the margin is a strong, raised rib, within which there is a row of large pores with strong ribs running upward as on the zocecial wall. The anterior pair of spines is inclosed within the ovicell. Two small colonies, the largest scarcely over 0.125 inch across, in- crusting algz, at 2 fathoms. In spite of the small size nearly every cell is adult, with a complete ocecium. I am a little uncertain as to the generic position of this species. Es- charella as amended by Levinsen (1909, p. 315) includes only forms without avicularia. In spite of this difference the species seems to fit better into Escharella than into any related genus. The absence of zocecial and ocecial pores, except the marginal ones, the well-developed vestibular arch, the very thin operculum, the character of the orifice and peristome, and the nature of the jointed spines, seem to locate the species in Escharella. In many respects it resembles E. diaphana MacGillivray, as figured by Levinsen (I. c.), except for the presence of the avicularia and the strong development of the ribs, which are wanting in diaphana. The Bryozoa of the Tortugas Islands, Florida. 205 GENUS SCHIZOPORELLA Hincks, 1880. Schizoporella unicornis (Johnston). JOHNSTON, 1847, p. 320 (Lepralia unicornis).—DESOR, 1848, p. 66 (Lepralia vari- olosa).—LEIDY, 1855, p. 10 (Escharina variabilis).—SMITT, 1873, p. 44 (Hip- pothoa isabelleana).—VERRILL AND SMITH, 1873, p. 713 (Escharella vari- abilis).—VERRILL, 1875), p. 41 (Hippothoa variabilts); ibid., p. 41, pl. 11, fig. 1 (Hippothoa reversa, n. sp.); 1878, p. 305 (H. variabilis); 1879b, p. 193, and 1880, p. 30 (Escharina isabelliana d’Orbigny, E. reversa Verrill and E. ansata Gray); 1900, p. 592 (? Schizoporella isabelliana).—LEVINSEN, 1909, Pp. 323.—OSBURN, I9QI2, p. 236. Abundant from low water to 10 fathoms, in the drift and growing profusely on the piles of docks, incrusting shells, and coral rocks, and rising into branched, finger-like processes about worm-tubes and the stems of hydroids, or occasionally growing free. Masses nearly as large as a man’s head were dredged. The color varies from white or pale pink or purple in young, rapidly growing colonies, to a dark purplish red or sometimes nearly black. Colonies 1.5 inches in diameter were found incrusting the bottom of a skiff that had been in the water only from May 1 to June 23. Smitt recorded the incrusting form ‘‘ growing in a single layer of whitish and yellowish hue on Oculina and Nullipora at the depth of 42 fathoms,” and, ‘‘a more compound colony, composed of several concentric layers of a purplish-blue tint, around an axis of foreign matter,” at a depth of 17 fathoms. Schizoporella floridana n. sp. (Figs. 17 and 18.) Primary characters of the zocecium much like those of S. unicornis (Johnston) but with a slightly more V-shaped sinus; avicularia situated far forward, often in advance of the middle of the orifice, and with the mandible usually strongly curved toward the zocecial axis; large vicarious avicularia with long-pointed, straight mandibles on hemispherical cells raised high above the surface of the colony. The naked-eye appearance of the colonies is very similar to that of S. unicornis (Johnston) and the general characters of the zocecia are not very different from that species. It does differ remarkably, however, in the presence of the large, vicarious avicularia, which are sometimes very abundant. These are raised high above the surface of the colony as rounded, knob-like prominences; the mandibles, while broad at the base, are very much narrowed and elongated distally. These are turned in all directions on the colony without any apparent arrangement. Schizoporella ampla Kirkpatrick, from Mauritius, and S. viridis Thornley, from the Red and Mediterranean Seas, also have vicarious avicularia. The ordinary avicularia are situated at the side of the orifice and the mandibles are curved toward the axis of the zocecium so as to nearly con- form to the curve of the margin of the orifice. There is much variation in this avicularium and the mandible is occasionally straight instead of curved, and it may be directed backward or sideways. It is usually situated well forward near the anterior border of the orifice, but may be placed farther 206 Papers from the Marine Biological Laboratory at Tortugas. back, occasionally behind the orifice. The zocecia are swollen about as in unicornis, the punctures are similar to that species, and the form of the orifice (with the sinus) might readily pass for that species. No oral spines have been observed. In the very young stage a low, raised margin separates the cells, but this is soon obscured by the secondary calcification. An umbo is developed below the orifice, apparently only by secondary calci- fication. In young colonies, where the vicarious avicularia are not yet developed, this species may readily be mistaken for S. unicornis, but the more anterior position and the curved form of the avicularia will serve to distinguish it even in this condition. The ocecium differs from that of any Schizoporella with which I am acquainted. It is comparatively small, very short (about two-thirds as long as wide), very high and prominent, smooth, and imperforate. This well-marked species, which appears to be undescribed, was dredged in 15 to 18 fathoms, incrusting shells and hard sponges, and, in one case, forming an erect nodular growth nearly an inch in height and about half an inch in diameter around some foreign matter as a center. The color ranges from pure white to dark purplish-red. Fic. 17.—Schizoporella floridana n. sp. Details of young zocecium. ‘ é Fic. 18.—The same. Highly calcified condition, showing ovicells and the long, independent avicularia mounted on large, swollen cells. Schizoporella sanguinea (Norman). NORMAN, 1868, p. 222 (Hemeschara sanguinea).—JELLY, 1889, p. 233, synonymy. Taken at 15 fathoms on a Vermetus shell; one colony with ovicells. Reported by Smitt from Pourtales’s collections at 60 fathoms, southwest of Tortugas. The Bryozoa of the Tortugas Islands, Florida. 207 Schizoporella biaperta (Michelin). MICHELIN, 1841-2, p. 330 (Eschara biaperta).—Smitt, 1873, p. 46 (Hippothoa biaperta) and p. 47 (Hippothoa divergens n. sp.).—VERRILL, 1875), p. 41 (Hippothoa biaperta); 1878, p. 305, 1879), p. 30, and 1880, p. 193 (Escharina biaperta).— HINCKs, 1880, p. 258 (S. biaperta var. divergens).—NORMAN, 1909, p. 303 (S. biaperta var. divergens)—LEVINSEN, 1909, p. 323 (discusses and amends genus).—OSBURN, I9I2, p. 237. A few colonies of this species were taken at various depths from low water to 22 fathoms. The appearance is characteristic and the specimens compare very well with those from the Woods Hole region, but never reach such a luxuriant development. Recorded by Smitt (under Hippothoa biaperta) from 9 to 60 fathoms, and (under H. divergens new species) from 135 fathoms ‘forma typica,” and 120 fathoms “‘forma laxa.” Smitt’s “H. divergens”’ has been ranked by Hincks and Norman as a variety of biaperta, and recorded by them from the British Isles and Madeira. All specimens examined by the writer belong to the typical biaperia. Schizoporella spongites (Pallas). PALLAS, 1766, p. 45 (Eschara spongites).—SMutT, 1873, p. 42 (Hippothoa spongites).— VERRILL, 1900, p. 592 (Hippothoa or Schizoporella Spongites).—LEVINSEN, 1909, p. 324 (full discussion of the species). One of the most characteristic species of the region; abundant from low water to 18 fathoms; incrusting shells, coral, rocks, and growing on the surface of harder sponges; also on piles. The very large ocecia serve to distinguish this species at a glance from any related forms. In life the color varies from translucent white or yellow to bright brick-red. Recorded by Smitt from Pourtales’s collections from 13 to 35 fathoms and by Verrill (1. c.) from Bermuda. Levinsen (I. c.) records it from St. Thomas and St. John, West Indies, and from Malacca (with some differences). GENUS ESCHARINA (PART) Milne-Edwards, 1838. Escharina pesanseris (Smitt). SMITT, 1873, p. 42 (Hippothoa pes anseris).—JELLY, 1889, p. 141 (Mastigophora duter- tret var. pesanseris).—WATERS, 1909, p. 169 (Schizoporella pesanseris).— NORMAN, 1909, p. 302.—LEVINSEN, 1909, p. 326 (discusses genus, amended, and species fully). One small dead specimen, composed of about a dozen zocecia, was taken at a depth of 8 fathoms. The vibracula characteristic of the species were broken off. Ocecia present. Smitt described this species from ‘‘two small colonies growing on a Nullipora, west of Tortugas” at 42 fathoms. It occurs also at Madeira in 70 fathoms (Norman), the Island of Mauritius (Kirkpatrick), the Red Sea (Waters), and Siam (Levinsen). 208 Papers from the Marine Biological Laboratory at Tortugas. Genus MICROPORELLA (PART) Hincks, 1877. Microporella ciliata (Pallas). PALLAS, 1766, p. 38 (Eschara ciliata).—SmITT, 1873, p. 26 (Porellina ciliata).—PACKARD, 1867, p. 270 (Lepralia ciliata)—VERRILL, 1879), p. 29 (Porellina ciliata); 1875¢, p. 53; 1880, p. 190, and 1879, p. 29 (Porellina stellata for a form of this species). —OSBURN, I912, p. 233 (M. ciliata), and p. 234 (the variety stellata Verrill).—LEVINSEN, 1909, p. 328 (discusses genus). From 5 to 18 fathoms on shells, corals, etc. One fine colony growing on an egg case of a large mollusk. Smitt records it at various depths from 7 to 60 fathoms. It is one of the most widely distributed of all bryozoa and occurs in all oceans. Levinsen (I. c.) places this genus in family Escharellide. GENUS SMITTINA NORMAN, 1903. Smittina trispinosa (Johnston). JOHNSTON, 1838, p. 280 (Lepralia trispinosa).—DAwWSON, 1859, p. 256 (Lepralia trispinosa).—PACKARD, 1867, p. 67 (Lepralia trispinosa).—SMITT, 1873, p. 59 (Escharella jacotint), and p. 60 (E£. spathulata).—VERRILL, 1879), p. 31; 1880, p. 195 (Mucronella jacotini); 1875a, p. 514, 1878, p. 305, and 1879), p. 30 (Discopora nitida).—WHITEAVES, 1901, p. 106 (Smittia trispinosa).— NORMAN, 1903, p. 120 (Smittina nom. nov. for Smiitia preoccupied in Dip- tera).—OSBURN, I9I2, p. 246 (Smittia trispinosa). One of the most abundant and characteristic species in the Tortugas region from low water to 12 fathoms. A cosmopolitan species and dis- tributed all along the Atlantic coast northward to the Arctic region. Smitt recorded it from 13 to 44 fathoms in Florida waters. Smitt states (p. 60) ‘“‘For the peculiarity of the development of the avicularia, the Floridan form, as a distinct variety, I propose to be named Escharella spathulata.’’ But variation in the secondary characters of this species here as in many other regions occurs to a bewildering extent. The typical form with the large, pointed avicularia, common in northern waters, is rare, but these are occasionally observed. One beautiful colony growing on the under side of Cupularia guiniensis at 10 fathoms is an almost pure example of Smitt’s spathulata. Even the smaller, triangular avicularia figured by Smitt (pl. x, fig. 200) are not represented, but in a few cases very elongated pointed avicularia occupy the place of the long-spatulate form figured by Smitt. Another colony found in the drift is more like Verrill’s nitida in the character of the avicularia, which are small, short- pointed, and short oval in form, very much like specimens from the southern New England coast. Ina few cases very large spatulate avicularia, as long as the whole zocecium, were observed. In one colony short-triangular avicu- laria were rather regularly disposed in pairs below the orifice with the man- dibles pointing outward. Occasionally the avicularia are almost wanting from a colony, a condition seen in many New England specimens that bear evidence of very rapid growth in a single layer. The ocecia vary much with the state of calcification. In younger stages they are coarsely and sometimes very irregularly punctured. This layer may be entirely covered by secondary calcification, when the ocecia present The Bryozoa of the Tortugas Islands, Florida. 209 a smooth, shining appearance or are variously roughened. The secondary layer usually grows irregularly over the primary layer, but in some cases it forms a rather regular raised border with a rounded area of the primary layer exposed on the top. This form resembles closely the variety protecta Thornely, as figured by Waters (1909, pl. 17, fig. 5), but the large avicularia are evenly rounded at the tip instead of being divided into points. The peristome also varies greatly. In some cases it is scarcely raised; in others it takes the form of a tall, broad, median projection, often forked at the tip and projecting so far over the orifice as to nearly obscure it from above. Ill sorts of intermediate conditions are observed. The peristome also occasionally shows raised lappet-like projections at the side of the orifice, as seen commonly in speci- mens from Massachusetts (Osburn, 1912, pl. xxvut, fig. 65a, 66). In spite of all these variations in secondary characters there is a remarkable con- stancy in the primary orifice, denticle, oral spines, and the primary zocecial walls. Another variation, and one which I am at a loss to explain, is this: Some colonies consist of zocecia, the secondary characters of which are all of one type or with little variation, while other colonies will show not only zocecia representing two or more of the so-called varieties, but all sorts of intergradations as well. GENUS LEPRALIA (PART) Johnston, 1847. Lepralia audouinii (d’Orbigny). D’ORBIGNY, 1850-2, p. 401 (Cellepora audouinii).—SmitTT, 1873, p. 56 (Escharella audouinit). This species, which is known from the Red and Mediterranean Seas, occurs at the Tortugas from low water to 10 fathoms. Colonies taken from the bottom of a skiff which had been in the water from May 1 to June 23 were an inch in diameter, with ocecia containing embryos. In life the colonies are translucent white, but the ocecia often appear light pink when embryos are present. Smitt’s record was based on a single colony growing on a nullipore at 37 fathoms, west of Tortugas. Lepralia porcellana Busk. BuskK, 1860, p. 283, pl. 31, fig. 3.—SmiTT, 1873, p. 62 (Lepralia cleidostoma).—NORMAN, 1909, p. 305, Synonymy. Taken at 5 to 15 fathoms on shells and on other bryozoa, such as Steg- anoporella magnilabris and Holoporella turrita. The key-hole form of the orifice, the pointed avicularia with elongate mandibles, and the absence of pores except occasional ones at the margin serve to distinguish the species readily. Odcecia present. In one case there is a vicarious avicularium, some- what larger than the usual type, mounted on a smooth, swollen chamber. A low, smooth, umbonate process is sometimes present below the orifice. Recorded by Smitt from Pourtales’s collections from 30 to 120 fathoms. 15 210 Papers from the Marine Biological Laboratory at Tortugas. Lepralia uvulifera n. sp. (Figs. 19 and 20.) Zocecia small, the surface shining and covered by small, smooth knobs. Pores are wanting except for 3 to 5 large, marginal pores. Usually these are placed as follows: One on either side near the orifice, one on either side about half-way back, and a single pore at the posterior end. The orifice is slightly broader than long, evenly rounded anteriorly, nearly straight on the posterior border, hinge teeth scarcely noticeable. Peristome slightly raised anteriorly and beset with about six slender spines. Posteriorly the peristome rises into a strong, mucronate process, which in complete calci- Fic. 19.—Lepralia uvulifera, n. sp. ; Fic. 20.—The same. Portion of colony showing at the right. the orifice of a young zocecium. fication is divided at the tip into three sharp spines. A small avicularium is present on one zocecium, situated somewhat behind the orifice near the lateral border, with the pointed mandible pointing backward and outward. The ocecium is comparatively large, heavily calcified, and covered with knobs like those of the zocecium; a high umbonate process rises on the top, and there is a single large pore on either side of the oceciostome near the base; the ocecial orifice is wide and high, with a calcified projection extending nearly vertically downward into the orifice and bearing a close resemblance to the uvula of the human palate, except that it is a little more truncate at the tip. One small colony of perhaps three dozen zocecia incrusting the under side of a Cupularia guiniensis at 10 fathoms. This species is certainly closely related to L. watersi Calvet (1906, p. 412, pl. xxvul, fig. 11) from the Cape Verde Islands, but it differs in having the umbo strongly trifid instead of merely pointed; in the more open form of the ovicell, which partially incloses the anterior part of the peristome, instead of being placed in front of it as in L. watersi; and also in lacking the complete lower border of the oceciostome shown in the figure of L. watersi. Calvet’s figure also shows the oral spines situated slightly outside of the oral border, while in uwvulifera the spines are on the border. The Bryozoa of the Tortugas Islands, Florida. 211 Lepralia cucullata Busk. Busk, 1854, p. 81.—WATERS, 1909, p. 150, synonymy and references.—JULLIEN ET CALVET, 1903, p. 141 (Schizoporella cucullata). Common in shallow water at the Tortugas. In life the color of the colony is dark brownish-purple, sometimes almost black, and the polypides in the young zocecia at the edge of the colony have a fine carmine color. A number of colonies more than an inch in diameter were found on the bottom of a skiff that had been in the water only from May I to June 23. The generic position of this well-known species is very unsettled. A majority of authors have agreed in placing it in Lepralia, others in Smittia, and still others in Schizoporella. The reason for this difference of opinion lies in the general simplicity of the zocecia, which lack characters usually considered as diagnostic. The form of the orifice is somewhat intermediate between that of Lepralia and Schizoporella. There is a very shallow curve on the posterior border, which may be interpreted as a sinus, as Calvet has done (l. c.), or as merely the space between the hinge denticles. In the absence of any means of settling the controversy, I place the species tenta- tively in Lepralia, in which genus it was described by Busk. Distribution: Mediterranean, Red, and Arabian Seas; Azores Islands; Cape Verde Islands; South Africa, and Cape St. Lucas, Lower California. Not before noted on the Atlantic side of America. Lepralia rostrigera (Smitt). SmitTT, 1873, p. 57 (Escharella rostrigera).—WATERS, 1885, p. 298; 1887, p. 61.—JELLY, 1889, p. 126 (under L. depressa). The Escharella rostrigera of Smitt has been made a synonym of L. depressa Busk by Miss Jelly (1. c.), but in the opinion of the writer it repre- sents a distinct species. Smitt recognized a relationship between the species, but indicated that the differences are constant. The study of material taken by me at the Tortugas confirms this view. The form of the zocecial aperture is quite different, being well rounded at the posterior mar- gin, as figured by Smitt, and there is never more than a faint indication of a constriction of the sides. The avicularia are characteristically placed far forward, alongside of the anterior part of the orifice, or frequently entirely in front of it. Busk’s figure of L. depressa (1854, pl. 91, figs. 3 and 4) shows no indication of pores. In rostrigera there are numerous small pores in the frontal wall in younger stages, and large, marginal pores are present in all stages. The larger zocecia with the broad aperture described and figured by Smitt (pl. x, fig. 205) are occasionally present in my specimens. Smitt’s specimens were taken by Pourtales in 35 and 43 fathoms. Mine were taken at 10 and 15 fathoms on coral and shells. A fine colony at Io fathoms incrusted the base of a living Cladocora arbuscula. Lepralia contracta Waters var. serrata Osburn. WATERS, 1899, p. I11.—NORMAN, 1909, Pp. 306.—OSBURN, I9I2, p. 242 (Lepralia serraia). Several colonies incrusting shells at 5 to 18 fathoms. This variety was 212 Papers from the Marine Biological Laboratory at Tortugas. described by me as Lepralia serrata n. sp., from the Woods Hole region (1912, p. 242, pl. 26, fig. 57). After a careful study of the Florida specimens I am of the opinion that this can be only a variety of L. contracta Waters, described from Madeira (1899, p. 11). Waters’s description is brief and his figures small, but Norman (Il. c., pl. 41, fig. 56) has refigured the species. The differences seem to be largely in the secondary calcification, which appears to be much greater in American specimens; also there are six oral spines in Madeira specimens, while usually only four are to be observed in American material. The avicularia in Florida specimens are much more abundantly developed and are frequently raised high above the front of the zocecium on small mammillary processes. Also, the lateral oral denticles are much more prominent than they appear in the figures of Madeira specimens and are strongly bifid; however, Norman figures one such bifid denticle. In one of my speci- mens the spatulate avicularia are extremely elongated, fully as long as the zocecium, and, while usually directed forward, are sometimes reversed in position. The zocecial wall appears to be much more thickened than in the Madeira specimens, to judge by the figures of Waters and Norman. Not- withstanding these differences I believe they are closely related. The form of the ocecium with the peculiar membranous area and the serrated char- acter of the primary zocecial orifice are identical and are so unique that I believe them to indicate the same species. Specimens from southern New England are much more highly calcified than Florida specimens, which seem in some respects to be intermediate between those from New England and Madeira. Lepralia edax (Busk). Busk, 1859, p. 59 (Cellepora edax).—SmiITT, 1873, p. 63 (L. edax forme typica and calcarea).—HINCKS, 1880, p. 311.—VERRILL, 1909, p. 54. One colony taken at 18 fathoms, forming a cylindrical mass more than 0.25 inch in diameter and many layers in thickness, which originally sur- rounded a branched structure of some sort, and one colony taken at 8 fathoms incrusting a shell fragment. Smitt’s figures and description of this species are very complete. I have compared these specimens with one from the English Channel sent me by Dr. S. F. Harmer. English speci- mens are said to be found only on certain species of gastropod shells. While Florida specimens have not been taken in such situations, I can distinguish no differences of any importance. My Tortugas specimens belong to what Smitt calls the forma calcarea, but it can scarcely be considered a distinct variety. The large, pointed, vicarious avicularia described and figured by Smitt are abundantly distributed over the colony. The ovicell has a peculiar membranous area on the upper surface, a feature also possessed by L. contracta Waters, though in the latter species this area is much nearer the zocecial aperture. The Bryozoa of the Tortugas Islands, Florida. Die Smitt records the species from Pourtales’s collections from 49 to 79 fathoms. Verrill records it for the Bermudas in shallow water. Lepralia janthina (Smitt). SMITT, 1873, p. 63 (Lepralia edax forma janthina). This, I believe, must be separated from edax as a distinct species. The form of the orifice, a character to which Smitt gave undue importance, is similar, but in janthina the orifice is not only much larger, but relatively wider and shorter, with less space behind the hinge denticles. The avicu- laria are all pointed and are situated on a prominence behind the orifice with the mandible turned forward and frequently slightly outward. There are no vicarious avicularia. The color is a deep blue-black or violet, entirely different from any specimens of L. edax Ihave seen. The zocecia are consid- erably larger than those of L. edax. Ovicells are wanting. One colony 0.25 inch in diameter taken at 6 fathoms incrusting a shell. Smitt records one specimen from 13 fathoms on coral. The Cellepora janthina of Waters (1899, p. 14) from Madeira can not be the L. edax var. janthina of Smitt, as Norman (1909, p. 311) points out in renaming Waters’s species C. rotundora. GENUS PHYLACTELLA Hincks, 1880. Phylactella labrosa (Busk). Busk, 1854, p. 92 (Lepralia labrosa).— JELLY, 1889, p. 204.—NORMAN, 1909, Pp. 308. Dredged at 22 fathoms on shells and hard sponges. Ovicells are lacking. There is no median denticle on the proximal margin of the orifice, nor are the zocecia disposed in radiating lines, but form a solid crust. However, Norman (1. c.) has shown that both of these characters are variable. Phylactella collaris (Norman) var. aviculifera nov. (Fig. 21.) Norman, 1866, p. 204 (Lepralia collaris); 1909, p. 309.—JELLY, 1889, p. 203.—ROBERT- SON, 1908, p. 307. This species has not heretofore been recorded from the Atlantic side of North America, though Miss Robertson records it from southern Cali- fornia, and it is found on the Euro- pean side of the Atlantic Ocean as far south as the Madeira Islands. At the Tortugas, growing on shells at from I to 15 fathoms, occurs the variety which I have here called aviculifera. In all re- 21 spects this appears to agree fully Fic. 21.—Phylactella collaris (Norman) aviculifera, var. with P. collaris, except that there —_20¥;,,, Viewed partly from/n front and showing avic is a rather large avicularium situ- ated within the collar immediately below the orifice, with a triangular mandible pointing upward, or very rarely sidewise. This appears on every zocecium of the colonies studied. Norman has described a rounded avicu- 214 Papers from the Marine Biological Laboratory at Tortugas. larium on the lower lip of occasional zooecia of L. labrosa from Madeira (1909, p. 309). GeENus CELLEPORA Linné, 1767. Cellepora dichotoma Hincks. HINcKs, 1862, p. 304.—SMITT, 1873, p. 53, pl. IX, fig. 193-198 (C. avicularis).—]JELLY, 1889, p. 51, gives synonymy and also, p. 46, includes Smitt’s references in C. avicularts Hincks.—(?) VERRILL, 1878, p. 305 (C. avicularis); (?) 1901, p. 54 (C. avicularis). Two well-developed specimens, 0.5 inch or more in height, taken at a depth of 10 fathoms; one was attached to a hydroid stem. There can be no doubt that these specimens belong to C. dichotoma instead of C. avicularis, as I have compared them with authenticated material from England. Smitt’s figures, which he refers to C. avicularis, undoubtedly are C. dicho- toma, as Hincks (1880, p. 404) has already pointed out. Smitt indicates the bathymetrical distribution as from 9 to 111 fathoms. Verrill’s records for C. avicularis at Fort Macon, North Carolina (1878), and Bermuda (1901) in all probability apply to dichotoma, though it is possible that the former should be C. americana Osburn (1912, p. 238), which Verrill at another time (18798) listed as avicularis. C. americana and C. dichotoma have been taken at Beaufort, North Carolina, by the writer. Cellepora verruculata Smitt. SMITT, 1873, p. 50.—Busk, 1884, p. 150 (Escharoides verruculata).—JELLY, 1889, p. 60.—CALVET, 1906, p. 444. A number of colonies taken on shells and in similar situations from low water to 15 fathoms. Smitt described the species from a single specimen taken by Pourtales west of the Tortugas in 42 fathoms. Recorded by Busk (1. c.) off Heard Island at 75 fathoms, and by Calvet (I. c.) for the Mediter- ranean Sea, ‘‘ Naples, cétes de Corse, Cette.” The zocecia undergo a remarkable change in appearance with advancing calcification. GENUS LAGENIPORA Hincks, 1877. Lagenipora ignota Norman. NORMAN, 1909, p. 3009, pl. 42, figs. 10-13. A colony taken at Tortugas in 12 fathoms on a dead colony of Mucronella bisinuata and another at 10 fathoms on a Vermetus shell seem to be identical with Norman’s species. They differ in not rising into rounded branches, but the small size of the colonies easily accounts for this. The character of the zocecium, with the pair of oral avicularia, is identical, and the occia also. The large spatulate avicularia, said by Norman to be present in extraordinary numbers, are but sparsely represented. Hitherto known only from the Madeira Islands, where it was taken at 70 fathoms (Norman). The Bryozoa of the Tortugas Islands, Florida. 215 Genus HOLOPORELLA Waters, 1909. Holoporella albirostris (Smitt). SMITT, 1873, p. 70 (Discopora albirostris) —Busk, 1881, p. 346 (Cellepora albirostris).— JELLY, 1889, p. 45 (references under Cellepora albirostris)—WATERS, 1885, Pp. 304; fate p. 68 (Cellepora albirostris); 1909, pp. 159-161 (Holoporella gen. nov.). This species, which is very characteristic of the region, occurs in abun- dance from low water to 15 fathoms. It grows on coral-rocks, corals, shells, and on the firmer sponges. Recorded by Smitt from Florida waters in 25 to 35 fathoms. It is readily distinguished from other species of this region by the presence of a towering, pointed white spine situated at one side of the proximal margin of the orifice. Except in the youngest stages the colony has a dark grayish or blackish color, against which the white spines stand out in sharp contrast. A very small avicularium is situated at the base of the spine, large spatulate avicularia are distributed over the colony, and vestigial, minute avicularia are occasionally present at the side of the orifice. Waters in 1909 (I. c.) established the genus Holoporella to include, for the most part, those species of the old genus Cellepora in which the sinus is lacking and which have a rounded or straight posterior margin of the orifice, and in which also the opercular muscles are attached near the border. Levinsen (1909, p. 347) erected a new family, Holoporellide, to include this genus, thus farther removing it from Cellepora. This separa- tion is based on the fact that in Holoporella the operculum is attached, sometimes by hinge teeth, to the lateral margins of the orifice, while in Cellepora the operculum is suspended at the edges of the sinus. Oral spines are often present in Holoporella, but are wanting in Cellepora. Rostra, with avicularia, may be present in both genera. Waters includes the following American species of the Florida region in Holoporella: H. (Discopora) albirostris (Smitt), H. (Lepralia) turrita (Smitt), and H. (Discopora) pertusa (Smitt). Levinsen adds H. (Discopora) advena (Smitt), which apparently completes the list of the described species of this region. Holoporella pusilla (Smitt). SMITT, 1873, p. 70 (Discopora albirostris forma pusilla).—Busk, 1884, p. 436 (Cellepora pusilla). In the ‘Floridan Bryozoa”’ Smitt describes this species as a form of his Discopora albirostris, stating that their relationship is ‘‘incontestably proved by the very same form of the zocecial aperture.” Busk (1. c.) objects to placing pusilla as merely a variety of albirostris, giving it as his opinion that the two seem to him to be quite distinct. Although Busk possessed no material of pusilla, he was evidently correct in forming this opinion. If the single specimen taken by me at the Tortugas is identical, as I believe it to be, with Smitt’s pusilla, it differs from albirostris in the entire absence of dark pigment; in the smaller size of the colony, the individual, and the orifice; in the presence of radiating lines of depressions and small 216 Papers from the Marine Biological Laboratory at Tortugas. wart-like prominences; in the smaller size of the ocecia; in the much slighter development of the rostrum, which never rises much above the avicularium; and in the more procumbent position of the rostrum, which projects forward over the orifice. The form of the orifice is, as stated by Smitt, very similar to that of albirostris, but (as Busk points out) this form is common to a number of species. Small hinge denticles are sometimes present. Smitt figures the species as having from 3 to 6 oral spines. There is scarcely any indication of these in the specimen in my possession. The oral avicularium is usually rounded, but occasionally the mandible is obtusely triangular. No other avicularia are present. One colony 0.25 inch in diameter incrusting a shell at low tide. Smitt records it from Pourtales’s collections at 9 to 60 fathoms. Holoporella magnifica n. sp. (Figs. 22 and 23.) A very coarse species with very large zocecia and orifices large enough to be distinguished readily by the naked eye. Taken at 10 fathoms incrust- ing a sponge and a very small colony on a dead shell at the same depth. ope: Fic. 22.—Holoporella magnifica n. sp. Young zocecium at edge of colony, showing manner of growth. Rostrum and rostral avicularium not yet developed. Fic. 23.—The same. Highly calcified portion near center of colony, showing one deeply immersed and three erect superficial zocecia, independent avicularia of two sizes, and the rostral avicularium. Zocecia large, procumbent at the growing edge, erect and irregular at the center of the colony. Imperforate except around the edge, where there are large pores, often coarsely cancellated between the cells in the middle of the colony. Very heavily calcified, even in young cells. The orifice is much as in H. albirostris, except that it is much larger. The peristome is very thick and broad, slightly raised all around the orifice (when ocecia are present it extends over them in later stages of growth). The rostrum is small and low, projecting so slightly over the orifice as to scarcely hide the posterior margin in any case. On one side of the rostrum is placed a large ww? The Bryozoa of the Tortugas Islands, Florida. 217 avicularium with an elliptical mandible, black in color. Sometimes the rostrum is so slightly developed that the avicularium appears to be placed transversely behind the orifice. The operculum is heavily pigmented with dark brown, especially at the edge. Large, spatulate avicularia with brown or blackish mandibles are abundantly distributed over the colony. There is a slight amount of the brownish pigment in the zoccial wall. In all the avicularia the pigment is confined to the mandible, which is thus strikingly contrasted with the colorless basal membrane. In the operculum also the color is limited strictly to the hinged portion and does not extend upon the base. The ocecia are, as in other species of this genus, shallow and widely open, and they are deeply immersed in the crust, except in the very young stage. In addition to the specimens mentioned above, a large flat specimen, 2 inches across and many layers in thickness, which was apparently growing unattached, was taken by Dr. Paul Bartsch off Biscayne Key, Florida. It has not been possible to identify these specimens with any known species. The large size of the zocecia and the orifice, the great thickness of the walls, even in young specimens, the straight proximal border of the orifice, the absence of a tall rostrum, and the large size of the oral avicularium, together with the elongate form of the mandible with its round tip in the vicarious avicularia, form a set of characters which serve to distinguish it. Holoporella turrita (Smitt). SMITT, 1873, p. 65 (Lepralia turrita).—WATERS, 1883, p. 438 (Smittia turrita); 1909 p. 161 (includes in Holoporella, new genus).—RIDLEY, 1881, p. 55 (Cellepora turrita).— JELLY, 1889, p. 253 (Smittia turrita). A very common species at 12 to 15 fathoms, growing on shells, corals, and the firmer sponges. Recorded by Smitt from Pourtales’s collections as not frequent, only a few specimens taken, at from 26 to 44 fathoms, growing on corals and nullipores. Color in life bright pink to brick-red. Above the spreading incrustations tise knob-like projections, usually 0.25 inch or so in diameter, sometimes much larger. The younger zocecia are separated by delicate, raised, white walls which are very conspicuous against the red color of the colony. The white points of the blunt spines are also strongly contrasted with the ground color. GENUS PETRALIA MacGillivray, 1869. Petralia bisinuata (Smitt). SMITT, 1873, p. 59 (Escharella bisinuata).—LEVINSEN, 1909, pp. 350-51. This well-marked species, which has not been noted outside of the Florida region, is common at the Tortugas on shells and sponges, from 10 to 18 fathoms. The color in life is bright vermilion. Smitt described the species from Pourtales’s collections at 9 and 19 fathoms. Levinsen (see above) has placed this species in MacGillivray’s genus Petralia and has erected a new family, Petraliide, to include this genus. 218 Papers from the Marine Biological Laboratory at Tortugas. CTENOSTOMATA. GENUS BOWERBANKIA Farre, 1837. Bowerbankia gracilis Leidy. LEIDY, 1855, Pp. 142.—HINCKs, 1877, p. 215 (Valkeria caudata); 1880, p. 521 (Bower- bankia caudata).—VERRILL AND SMITH, 1873, p. 709 (Vesicularia gracilis), and p. 710 (V. fusca).—VERRILL, 1880, p. 28 é Vesicularia gracilis).—LrEv- INSEN, 1894, p. 82 (Bowerbankia caudata).—OsBURN, 1912, pp. 253-245 (synonymy); 19124, p. 287. Specimens of this species, which do not seem to differ from Woods Hole material, were taken in the moat at Fort Jefferson and on the piles of the docks, attached to shells, or creeping over the sea-weed, and covered to such an extent by coral mud as to almost obscure them. No erect branches were noted. Many individuals show the caudate process of the variety caudata (Hincks). On the North American coast the species has been identified by the writer from Nova Scotia, Maine, Massachusetts, Connecticut, New Jersey, North Carolina, and Florida. Otherwise it is known from England and Denmark. It is quite probable that the genus Bowerbankia, as Waters (1910, pp. 241 and 244) suggests, must be fused with Zoobotryon, but as Waters himself keeps them separate pending more thorough studies of the ctenostomes, the writer has not the temerity to combine them. The only marked dis- tinguishing characters are the much greater softness and hyalinity of the stock and the absence of a creeping stolon in Zoobotryon. These characters will readily separate the one species of Zoobotryon from any Bowerbankia with which the writer is acquainted. GENuS ZOOBOTRYON Ehrenberg, 1831. Zoobotryon pellucidum Ehrenberg. EHRENBERG, 1831, pl. 111, fig. 10.—SONDER in Coll. Binder (Ascothamnion trinitatis).— WATERS, IQIO, p. 243. This species, first described from the Mediterranean, is common at the Tortugas, floating and attached, especially in the Fort Jefferson moat and about the piles of the dock. One colony 8 inches across was observed. The zocecia in all cases were much obscured by a layer of white coral mud adhering to the ectocyst. Not previously recorded for the Florida region. Sonder (see above) recorded it as a plant from the island of Trinidad, and according to Waters (see above) it is known from the Isle of Pines, South Australia, Zanzibar, the Red Sea, and the Cape Verde Islands. GENUS CYLINDRECIUM Hincks, 1880. Cylindrceecium giganteum (Busk). BuskK, 1856, p. 93 (Farrella gigantea).—HINCKS, 1880, p. 535. This species, which has been recorded from the British Isles, from various localities in the Mediterranean Sea, from the Red Sea, and from the Queen The Bryozoa of the Tortugas Islands, Florida. 219 Charlotte Islands, has not hitherto been noted on the American side of the Atlantic. It is abundant at the Tortugas from low water down to several fathoms, growing profusely over piles, shells, seaweed, and sponges. The zocecial wall in all cases was thickly covered with a layer of coral mud. Occasional individuals showed the swelling at the side of the zoceciostome which forms the ocecium. Embryos in various stages of development were present. Genus ANGUINELLA van Beneden, 1844. Anguinella palmata van Beneden. VAN La 1844, p. 58.—Busk, 1856, p. 95.—OSBURN, I9I2, p. 253, pl. XXVIII, g. 78. This species occurs rather sparingly among algz on the reefs in shallow water. It is very easily overlooked because its irregularly branched form gives it a marked resemblance to some of the smaller alge. The species is now known to occur on the American coast from the Tortugas to Buzzard’s Bay, Massachusetts (Osburn, 1912). At Beaufort, North Carolina, it is very abundant and grows much larger than at the localities mentioned above. It has not previously been noticed south of Charleston, South Carolina, where it was recorded by Busk (Harvey, coll.). GENus AMATHIA Lamouroux, 1812. ? Amathia goodei Verrill. VERRILL, I901, p. 329.—(?) SMITT, 1872, p. 4 (Serialaria sp. undet.). Several colonies, growing attached to piles, seem to belong to this species, which Verrill described from the Bermudas. Verrill described the branches as ‘‘thick, soft and flaccid’’ and the zodids as “‘ numerous, arranged in large, dense elongated clusters composed of several close rows, which often nearly or quite surround the stem, but are scarcely at all spiral.’’ BIBLIOGRAPHY. Aupoutn, J. V. 1826, Explication sommaire des planches des mollusques, des annélides, des crustacés, des arachnides, des insectes, des échinoderms, des ascides de L'Egypte et de la Syrie, par G. C. Savigny. Paris. BENEDEN, P. J. VAN. 1844. Recherches sur l’anatomie, la physiologie et le développe- ment des bryozoaires qui habitent la cOte d’Ostend. Nouveau Mémoires Académie Royale Belgique, t. xvi, 1845. Bruxelles. Bosc, L. 1802. Histoire naturelle des vers, vol. +: Busx, G,. saeshy Catalogue of the marine polyzoa in the collection of the British u seum. Chilostomata, I, 1852; part 2, 1854. 1856. Zoophytology. Quarterly Journal of Microscopical Science, vol. 4, pp. 93-96, pl. 95-96. —— 1859. The polyzoa of the Crag. Paleontographic Society. —— 1860. Catalogue of the polyzoa collected by J. Y. Johnson at Madeira, etc. anaaghd Journal of Microscopical Science, vol. 8, pp. pote pl. 30, 31. —— 1881. Polyzoa collected by Capt. H. W. Feilden in the North Polar dase aoe with descriptions of new species. Journal of the Linnean Society, Zoology, vol. 15, pp. 231-241, pl. 13. ——— 1884. Report on the polyzoa collected by H. M. S. Challenger, pt. 1, Cheilo- stomata. Vol. X, pt. XXX, pp. (XxIv) 1-216, pl. -xxxvi._ London. 1886. Idem, pt. 2, Cyclostomaia, Ctenostomaia, and Pedécellinea. Vol. xvut, pt. L, pp. (vit) 1-47, pl. r-x. CatvetT, L. 1906. Expédition Scientifique des Travailleur et du Talisman. Pp. 355- 495, pl. 26-30. Couc#, R. Q 1844. Cornish Fauna. Vol. 3. Dawson, J. W. 1859. Geological Survey of Canada for 1858. lets PP. 255-257. Dersor, E. 1848. Ascidioidian polyps or bryozoa (from Nantucket). Proceedings of the Boston Society of Natural History, vol. 3, pp. 66-67. ERRENBERG, C. ed 1831. Symbol Physice, pt. 2, Evertebrata. Pages not numbered, pl. 3, fig. ro. FLEMING, J. 1828. A history of British animals, exhibiting the descriptive characters and systematical arrangements of the genera and Sieg: of quadrupeds, birds, reptiles, fishes, Mollusca, and Radiata of the United Kingdom. First ed., 8vo, Zoophytes, pp. 505-554. Edinburgh. Gasp, W. M., and Gro. H. Horn. 1862. Monnans of the fossil polyzoa of the second- ary and tertiary formations of North America. Journal of the Academy of Natural Sciences of Philadelphia, vol. 5, pt. 2, pp. 111-179, pl. 19-21. Gray, J. E. 1848. List of British Animals in the British Museum, part 1. HARMER, S. F. 1891. On the British species of Crista. Quarterly Journal of Micro- scopical Science, n. s., vol. 32, pp. 127-181, pl. 12. 1900. A revision of the Genus Steganoporella. Quarterly Journal of Micro- scopical Science, n. s., vol. 43, pp. 225-297, 3 12-13. Hincks, T. 1862. A catalogue of the zoophytes of South Devon and South Cornwall. Annalsand Magazine of Natural History, ser. 3, vol. + Pp. 303-310, pl. 12. 1877. On polyzoa from Iceland and Labrador. Annals and Magazine of Natural History, ser. 4, vol. XIX, pp. 97-112, pl. x, x1. London. —— 1880. British marine polyzoa. Vol. 1, 601 pages of descriptive matter; vol. 1, 83 pl. to accompany text. London. ——— 188o0a. Contributions toward a general history of the marine Polyzoa. Annals and Magazine of Natural History, ser. 5, vol. 6, pp. 69-91, pl. 9-11. ———_ 18Sos. Same title, pt. 2 and 3, same journal, ser. 5, vol. 6, pp. 376-384. —— 1881. Same title, pt. 4, same journal, ser. 5, vol. 7, pp. 147-162, pl. 8-10. 188ra. Same title, pt. 8, same journal, ser. 5, vol. 8. 1887. Critical notes on the polyzoa. Same journal, ser. 5, vol. 19, pp. 150-164. lexus, es. &: wie i Pe synonymic catalogue of the recent marine bryozoa, 322 p. ondon. Jounston, G. 1838. History of British zoophytes. London. Bryozoa confused with other groups under Ascédioida. Pp. 238-324, pl. XXIX—XLII. 1847. Idem, 2d ed. Polyzoa, pp. 253-406, pl. XLVI-LXXIV. ——S—E ee eeeeeSE—E———=—e elle eee ee ee EEE The Bryozoa of the Tortugas Islands, Florida. 221 JuLuren, J. 1881. Note sur une nouvelle division des Bryozoaires Cheilostomiens. Bul- letin Société Zoologique du France, vol. 6, pp. 271-285. 1888. Mission Scientifique du Cap Horn. Bryozoaires. Tome 6, Zoologie, pp. I-92, pls. I-15. JULIEN, J., et L. CALVET. 1903. Bryozoaires provenant des campagnes de L’Hirondelle. Résultats des campagnes scientifique du Prince de Monaco, fasc. XXIII, pp. 1-188, pl. i-xvit. Monaco. Lamarck, J. B. 1816. Histoire naturelle des animaux sans vertébres, vol. 1, 1° éd., Paris. Bryozoa scattered among “ Polypes.”’ LanpsporoucH, D. 1852. A popular history of British zoophytes. London. Polyzoa, pp. 265-386, pls. 15-20. Leipy, J. 1855. Contributions toward a knowledge of the marine invertebrate fauna of Rhode Island and New Jersey. Journal of the Academy of Natural Sciences of Philadelphia, ser. 2, vol. 3 (Polyzoa on pp. 9-11). Levinsen, G. M. R. 1894. Mosdyr (Polyzoa eller Bryozoa). In: Schiodte, J. C., Zoologica Danica, 4de bd., Iste afd., pp. 1-105, pl. 1x. Kjobenhavn. 1909. Morphological and systematic studies on the cheilostomatous bryozoa. 364 pages, 24 plates. Copenhagen. LINNE, C. 1758. a stema nature, ed. 10, vol.1. Lithophyta and zoophyta, pp. 789-821. olmiz. 1767. Idem, ed. 12, vol. 1, pt. 2. Lithophyta and zoophyta, pp. 127-1337. Holmiz. MacGitiivray, P. H. 1869. Descriptions of some new genera and species of Australian Polyzoa, etc. Transactions and Proceedings of the Royal Society of Victoria, part 2, vol. 9, pp. 126-148. MIcHELIN, H. 1841-2. Iconographie zoophytologique ... des polypiers fossiles de France, . . . XII, 348 p., atlas of 79 pl. Paris. Norman, A. M. 1864. On undescribed British hydrozoa, actinozoa, and _polyzoa. Annals and Magazine of Natural History, ser. 3, vol. 13. 1903. Notes on the natural history of East Finmark. Annals and Magazine of Natural History, ser. 7, vol. 12, pp. 87-128, pl. 8-9. 1909. The Polyzoa of Madeira and neighboring islands. Journal of the Linnean Society, vol. Xxx, pp. 275-314, pl. 33-42. Orpicny, A. dD’. 1839. Voyage dans l’Amérique méridionale. Vol. 5, pt. 4, Bryozoa, pp. 7-23, pl. 1-10. Paris. 1850-2. Paléontologie frangaise, Terrains crétacés. Vol. 5, pp. I-1191, pl. 600-800. Ospurn, R. C. 1912. The bryozoa of the Woods Hole region. Bulletin of the United States Bureau of Fisheries for 1910, vol. 30, pp. 201-266, pls. 18-31. 1912A. Bryozoa from Labrador, Newfoundland, and Nova Scotia, collected by Dr. Owen Bryant. Proceedings of the United States National Museum, vol. 43, pp. 275-289, pl. 34. ; Packarp, A. S. 1863. List of animals ‘dredged near Caribou Island (Labrador). Cana- dian Naturalist and Geologist for 1863, pp. 406-412. 1867. Invertebrate fauna of Labrador and Maine. Proceedings of the Boston Society of Natural History, vol. 1, pp. 66-69. Pauias, P. S. 1766. Elenchus zoophytorum. Hage Comitum. Prerer, F. W. 1881. Eine neue Bryozoe der Adria, Gemellaria (?) avicularis. Jahres- ber. Westfialischen Prov.-Vereins, vol. 9, pp. 43-48, pl. I. PourtaLes, T. F. pE. 1867. Contributions to the fauna of the Gulf Stream at great depths. Bulletin of the Museum of Comparative Zoology, vol. 1, No. 6 (Bryozoa on pp. 106 and I10-II1I). ‘ Reuss, A. E. 1847. Die fossilen Polyparien des Wiener Tertiar-beckens. Wien. Ripvey, S. O. 1881. Account of the zoological collections made during the survey of H. M.S. Alert. Proceedings of the Zoological Society of London. ROBERTSON, ALICE. 1905. Non-encrusting chilostomatous bryozoa of the west coast of North America. University of California Publications, Zoology, vol. 1, No. 5, pp. 235-322, pl. 4-16. , 1908. The encrusting chilostomatous bryozoa of the west coast of North America. University of California Publications, Zoology, vol. Iv, No. 5, pp- 253- 344, pl. 14-24. Smitr, A. F. 1872-3. Floridan Bryozoa, collected by Count L. F. de Pourtales. Kongl. Svenska Vetenskaps-Akademiens Handlingar, pt. 1, 1872, in bd. 10, No. 11, pp. 1-20, taf. I-Iv; pt. 2, 1873, in bd. 1, No. 4, pp. 1-83, taf. I-XIII. Stockholm, SOLANDER, D. 1786. Natural history of many curious and uncommon zoophytes, col- lected from various parts of the globe by the late John Ellis, syste- matically arranged and described by the late D. Solander. London. 222 Papers from the Marine Biological Laboratory at Tortugas. Stimpson, W. 1853. Synopsis of the marine invertebrata of Grand Manan. Smith- sonian Contributions to Knowledge (Bryozoa, pp. 17-19, pl. 1). VERRILL, A. E. 1875. Brief contributions to zoology from the Museum of Yale College, No. 32. American Journal of Science and Arts, vol. 9 (Bryozoa on p. 414). 1875A. Idem, No. xxx. Results of dredging expeditions off the New England coast in 1874. Ibid., vol. x. Bryozoa, pp. 41-42, pl. m1. ——— 1878. In: Coues and Yarrow, Notes on the natural history of Fort Macon, North Carolina, and vicinity. Proceedings of the Academy of Natural Sciences of Philadelphia. List of polyzoa by Verrill on pp. 304-305. —— 1879. Preliminary check-list of the marine invertebrata of the Atlantic Coast from Cape Cod to the Gulf of St. Lawrence. Pp. 28-31. (Published privately, New Haven, Connecticut, April 1879.) —— 1879A. Brief contributions to zoology from the Museum of Yale College, No. xLi11, Notice of recent additions to the marine fauna of the eastern coast of North America, No. 6. American Journal of Science and Arts, vol. XVIII, pp. 52-54. New Haven. —— 1880. Notice of recent additions to the marine invertebrata of the Atlantic coast of America. Proceedings of the United States National Museum, vol. 2, 1879 (published 1880). Polyzoa, pp. 188-196. —— 1900. Additions to the Tunicata and Molluscoidea of the Bermudas. Zoology of the Bermudas, vol. 1 (Bryozoa on pages 592-594, figs. 4 and 6 of pl. 20). —— 1901. Additions to the fauna of the Bermudas from the Yale Expedition of 1901. Zoology of the Bermudas, vol. 1 (Polyzoa on p. 54). —— 1g01A. Review of recent papers relating to the fauna of Bermuda. American Journal of Science, vol. 11 (on page 29 in a footnote is a description of Amathia goodei n. sp.). VERRILL, A. E., and S. I. SmirH. 1873. The invertebrate animals of Vineyard Sound and adjacent waters. Report of the Commissioner of Fish and Fisheries for 1871-2. (Bryozoa, pp. 707-714 and p. 747.) Washington. Waters, A. W. 1883. Fossil chilostomatous bryozoa from Muddy Creek, Victoria, etc. Quarterly Journal of the Geological Society, vol. 39, pp. 423-443, pl. 12. 1885. Chilostomatous bryozoa from Aldinga and the River-Murray Cliffs, South Australia. Same journal, vol. 41, pp. 280-310, pl. 7. —— 1887. On tertiary chilostomatous bryozoa from New Zealand. Same journal, vol. 43, pp. 40-72, pls. 6-8. —— 1898. Observationson Membraniporide. Linnean Society Journal, Zoology, vol. XXVI, pp. 654-693, pl. 47-49. London. ——— 1899. Bryozoa from Madeira. Jour. Roy. Micros. Soc., pp. 6-16, pl. 3. —— 1904. Resultats du voyage du S. Y. Belgica. Bryozoa, pp. 1-103, pl. I-9. ——— 1906. Bryozoa from Chatham Island and d’Urville Island, New Zealand. Annals and Magazine of Natural History, ser. vit, vol. XVII, pp. 12-23, pl. I. 1907. Tubucellaria: Its species and ovicells. Linnean Society Journal, vol. xxx, pp. 126-133, pl. 15, 16. 1909. Reports on marine biology of the Sudanese Red Sea, etc., xm, Bryozoa Pea oat part 1, Linnean Society Journal, vol. xxx1, pp. 123-181, pl. 10-18. 1910. Same title, xv, Bryozoa Cyclostomata, Ctenostomata, and Endoprocta, pt. 11, same journal, vol. XxxI, pp. 231-256, pl. 24, 25. WHITEAVES, J. F. 1901. Catalogue of the marine invertebrata of Eastern Canada. Geological Survey of Canada, Ottawa. Polyzoa, pp. 91-144. Nek hea hi Peo 4 is ue jv ge faba lla cht AG i ao £.p a 4 ol | a pAR Le Lwe dd t hdd adda “vippel? , Tn ‘ YL EU EL aps a sag * aa cal Awa “<7 4P 4 Lieb hk -9,96%- inte? eaaleth Pao nl pateat! ppb rb -f/ Perea As pie i a7, 2 \ pz ) 4.44 LD ; a, aAK var ga male : Lee eAne ~ tty @: Ap, Aw. 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