..__—--l!-!!ll!ll!55“F—--fi!-.lll!gpl ERRATA November, 1958 chlichting, Harold E., Jr. Thr Role of Waterfowl nthe Dispersal of Algae. Unpublished doctoral Msis, Michigan State University. 1958. .208 ...greater variety was... line #6 .213 Purple Martin should not be classified as waterfowl. .249 Fetteroff-date entered twice Fassett .251 Irenee Marie Flor (drOp e) Egg (not Dis) 252 Kudo-date entered twice .256 Buschkarew-delete the second der Verbreitun RaorJournal of the ZEQAEE Botanical Society (not Indiana) 257 Savile-Vol.w# not included 259 Zacharias Uber, Schwimmvogel .._ THE ROLE OF WATERFOWL IN THE DISPERSAL OF ALGAE By 9" HAROLD EUGENE SC HLICHTING. JR. A THESIS Submitted to the School for Advanced Graduate Studies of Michigan State University of Agriculture and Applied Science in partial fulfillment of the requirements for the degree of DOC TOR OF PHILOSOPHY Department of Botany and Plant Pathology 1958 .2 1% a“? :9 7 ,5“ /3 THE ROLE OF WATERFOWL IN THE DISPE RSAL OF ALGAE By Harold Eugene Schlichting. Jr. AN ABSTRACT Submitted to the School for Advanced Graduate Studies of Michigan State University of Agriculture and Applied Science in partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY Department of Botany and Plant Pathology 1958 Approved ABSTRAC T Ideas expressed by most ecologists concerning the dispersal of micro-organisms by waterfowl have been largely based on assumptions or upon data from a few field collections. A series of controlled experiments was conducted at the W. K. Kellogg Bird Sanctuary on Wintergreen Lake. Kalamazoo County, Michigan. to determine the possible role played by waterfowl. Ducks were trapped and. in certain phases of the experiment, some were washed in a detergent. They were then placed in a water pen in Wintergreen Lake for periods of time varying from 15 minutes to 24 hours. After removal from the water pen. the ducks were exposed to the air for periods ranging from 15 minutes to 32 hours, either in an air cage or by being hung on a clothesline in a harness. A second series of experiments was conducted in which the ducks were placed in a mud pen. The ducks were restrained in a holding funnel while plastic boots filled with boiled pondwater were tied around their feet to remove any organisms present. Micro-organisms obtained in the boot washings were cultured in soil-water medium. Washings from the bills and feathers. the Contents from the gullets, and faecal material of some of the birds. ii Q and also washings of field-collected birds were cultured. All cul- tures were examined microscopically to determine the presence or absence of organisms. In some instances examinations were also made of the uncultured material. Controls were maintained by (1) sampling water taken from the water pen, while the ducks were there. to determine the micro- organisms present, (2) exposing the ducks and boiled pondwater to the air for the same period of time, and (3) observing cultures of unexposed boiled pondwater as used in the washings and uninoculated Culture medium to determine the presence or absence of micro- organisms. Environmental data such as humidity, wind velocity, air temperature, and sky conditions during the period of investigation were recorded. One hundred and six waterfowl. representing seventeen species. Were washed with boiled pondwater. Forty-one birds were used for the field data. whereas 23 ducks were used in the controlled experi- ments in 1955 and 42 in 1956. Viable organisms found on the Waterfowl were 87 species from the feet, 26 from the feathers, 2.5 fI‘om the bills, 14 from the gullet. and 8 from the faecal material. The modes of dispersal as well as the'nature of the aquatic eIl‘rironment determine what organisms are to be found in a giVen environment. Although often not considered, these modes are iii also important in explaining the distribution of aquatic micro- organisms throughout the world. iv PRE FAC E A Truism: Hardly any branch of natural history has been so neglected as that which treats of the various modes by which the different classes of organisms have become dispersed over the surface of the globe. Alfred R. Wallace (1893-1913) Quotation from Kew (1893, p. v of Preface) The primary objective in undertaking the following research was to demonstrate under what conditions algae might be dispersed by waterfowl and to study previously unknown factors through con- trolled experiments. This study was conducted in the hope that it might stimulate interest in problems of dispersal of micro-organisms and that it might be the basis for future research in this field to give us more insight into methods by which organisms are dispersed. No man is an island, entire of itself; every man is a piece of the continent, a part of the main. [Merton, 1957, p. 21] This seems especially true in ecological research. One can- not isolate himself from the work and ideas of others if he is to contribute to the advancement of knowledge. It is therefore with great respect and gratitude that those who have aided in advancing this research are acknowledged. Dr. Gilbert M. Smith of Stanford University was the first to arouse my interest in the dispersal of algae. and were it not for his encouragement the study would not have been initiated. I am also deeply indebted to Dr. G. W. Prescott of the De- partment of Botany and Plant Pathology and Dr. T. W. Porter of the Department of Zoology, Michigan State University, for their encour- agement, understanding. and guidance. I also wish to thank Drs. _ M. D. Pirnie of the Department of Fisheries and Wildlife, W. E. Wade 0f the Department of Natural Science, and G. P. Steinbauer of the Department of Botany and Plant Pathology for their helpful advice. Gratitude is also extended to Drs. I. W. Knobloch, P. H. Barrett, C. H. Nelson, and Messrs. D. L. Shull and D. K. Stewart, of the Department of Natural Science, Drs. D. E. Schoenhard of the Départment of Microbiology and Public Health. R. C. Ball of the Department of Fisheries and Wildlife, Michigan State University; A. D. Geis of Patuxent Wildlife Research Station, Laurel. Maryland; Drs. W- T. Edmondson, K. L. Osterud, and D. L. Ray of the Department of Zoology, University of Washington, Seattle; E. G. Pringsheim of Pflarizen-physiologischles Institut. Gottingen. Germany; W. J. Clench 0f the Museum of Comparative Zoolog at Harvard University; Drs. C' T. Black. T. J. Peterle, and Mr. Ray Schofield of the Michigan Department of Conservation Rose Lake Wildlife Experimental Station; Vi Mr. Thomas Graham of Thurso. Scotland; and Mr. W. H. Southworth, Farm Foreman at Michigan State University. For summer facilities I am indebted to the W. K. Kellogg Gull Lake Biological Station of Michigan State University, and R. D. Van Deusen, Director of the W. K. Kellogg Bird Sanctuary. Messrs. A1 England and Richard Cleaves aided in the construction of pens. traps, and other apparatus, for which I am very grateful. Gratitude is also extended to Mrs. H. V. Konkel of Detroit for aiding in the translation of De Guerne's French publication. I also wish to thank Mr. Edwin Wintermute of The Lansing State Journal for his encouragement and for critically reading this thesis. Lastly, I will be forever indebted to my- wife. Mary Southworth SChlichting. along with my relatives and friends. who make any diffi- cult task well worth doing. C HAP IV. C C HAPTER TABLE OF CONTENTS I. INTRODUC TION ........................ Origin of the Problem .................... Related Problems of Dispersal ............. Physical Agencies .................... Wind . Flooding and rain run- off ............. Water currents and floating objects ...... Biological agencies .................... Birds Insects Fish . Reptiles ......................... Amphibians ....................... Mammals ........................ Man. ooooooooooooooooooooooooooo II. HISTORY OF THE PROBLEM .............. III. HUNTERS' DATA SIEETS AND FIELD DATA .......................... Methods ........................... Field Collections ..................... Hunters' data sheet ................. Seattle. Washington ....... _ ........... Pine River. Michigan ................ St. Joseph River. Michigan ............ Port Sanilac. Michigan ............... IV. CONTROLLED EXPERIMENT. 1955 .......... Description of Wintergreen Lake .......... Materials Controls 00000000000000000000000000 ........................... 21 22 23 23 24 30 32 33 36 36 39 43 CHAPTER Page V. CONTROLLED EXPERIMENT. 1956 .......... 74 Modifications ........................ 74 Control Culture Flasks ................. 84 Winter Data ......................... 87 VI. DISCUSSION ........................... 190 VII. CONCLUSIONS ......................... 207 APPENDIXES ................................ 211 A. ADDITIONAL TABLES ................... 212 B. SUR LA DISSENIINATION DES ORGANISMES D'EAU DOUCE PAR LES PALMIPEDES .................. 236 BIBLIOGRAPHY . . ........................... 246 ix ea VIII. TABLE II. III. IV- VIII. LIST OF TABLES Birds Examined in the Hunters' Data Sheet Study. Fall. 1953 ............ Organisms Observed on Microscopic Examination of Washings from the Birds in the Hunters' Data Sheet Study . Fall. 1 953 .................... Data from Controlled Experiments with Classification of Organisms, 1955 ........ Organisms in Water Pen. 1955 .......... Classification of Organisms Observed in Uncultured Washings, 1956 ........... Data from Controlled Experiments (Water Pen) with Classification of Organisms. 1956 ..................... Data from Controlled Experiments (Miscellaneous) with Classification of Organisms , 1 956 .................. Data from Controlled Experiments (Mud Pen) with Classification of Organisms. 1956 .................... Data from Controlled Experiments (Control Flasks) with Classification of Organisms , l 956 .................. Feather Washings Planted November 2. 1956. with Classification of Organisms ......................... X Page 25 27 49 73 89 90 128 136 I46 182 TABLE Page XI. Feather and Other Washings Planted November 23, 1956, with Classification of Organisms . ............... 184 XII. Waterfowl Used for Washings .............. 213 XIII. Micro-organisms Found on Waterfowl Used in the Controlled Experiments .......................... 2 1 4 XIV. Planktonic Micro—organisms Taken from Wintergreen Lake during the Summer of 1956s-Qualitative Study ........... 218 XV. Environmental Factors in the Controlled Experiments. 1956 .............. 228 XVI. pH of Culture Flasks. 1956 ................ 232 XVII. Field Data Sheet for Duck Hunters ........... 235 xi LIST OF FIGURES Figure Page 1. Aerial View of Wintergreen Lake ............ 37 2. Funnel Trap .......................... 41 3. Water Pen ............................ 42 4. Air Cage . ............................ 44 5. Duck in Air Cage ....................... 44 6. Duck with Plastic Boots in Funnel ........... 45 7. Wintergreen Lake Showing Filamentous Algal Mat and Lily Pads with Boathouse in Background ......................... 75 8. Ducks in Water Pen ..................... 76 9. Ducks in Harnesses and Fingerbowl Hanging on Clothesline ................... 78 10- Ducks and Fingerbowl Covered with Cheesecloth Netting Hanging on Clothesline ..... 79 11 . Duck with Plastic Boots in Funnel. Side View ............................ 81 12-- Duck with Plastic Boots in Funnel. Rear View ............................ 82 13- Culture Flasks ......................... 83 14- Mud Pen ............................. 35 xii CHAPTER I INTRODUCTION Origin of the Problem While studying at the University of Michigan Biological Station in 1953, the writer became interested in the role waterfowl are as- sumed to play in the dispersal of algae and protozoa. The following quotations from Dr. G. M. Smith's text, 3112 Ereshwater Algae of the United States, called my attention to the fact that there was very little experimental evidence to support the generalizations made by various ecological investigators in respect to algal dispersal. Smith (1933. p. 11, 12) states: All discussions of the means by which alga [gig] are dis- Persed have been based upon general observations rather than a detailed study. and it is not definitely known whether algae are transported in a vegetative or in a resting stage. The importance of zygotes and resting cells has been greatly overemphasized in discussions on dispersal; it is very Probable that dissemination of vegetative cells is of far greater iImportance than that of resting cells. Streams assist in the dispersal of algae. but the two major agencies transporting algae fI‘om one locality to another are birds and the wind. Those [persons] who argue for transportation by birds hold that most of the algae are carried in half-dried mud adhering to the bird's 1 feet, but lodging of algae among the bird's feathers may be fully as important a factor. Transfer of plankton algae is brought about by migratory aquatic birds moving from one body of water to another. After realizing that a problem existed, the literature was searched to learn what investigations concerning methods of dispersal had been conducted. When it became clear that very little evidence was at hand, a plan of research was devised by which it could be learned whether waterfowl do play an influential role in the dispersal of freshwater algae. Attempts were made to develop several schemes Which would show empirically whether wind, flooding. rain run-off. water currents, floating objects, birds, insects, fish, reptiles, am- Phibians, mammals, and man were all involved in the dispersal of aquatic organisms. However, this thesis concerns only the role of waterfowl in the dispersal of algae. Related Problems of Dispersal Various agencies have been credited with dispersing micro- organisms throughout the world. .fiphysioal agencies Wind. - - There have been recorded in all periods of historic time. however, showers of one kind or another of animals and plants or their products-showers of hay. of grain. of manna. of blood. of fishes. or frogs, and even of rats. [McAtee. 1917, p. 217] Many researchers such as Gislen (1940 and 1948), Messikom- mer (1943). Hudson (1889). Beger (1927), and Huber-Pestalozzi (1937) have stressed the importance of air currents in the dispersal of micro-organisms. Gislen (1940, p. 22), Hudson (1889. p. 173), and Pennak (1953, P- 15) have indicated that the ability of an organism to form a light Spore or cyst will probably also explain the wide distribution of that Species and its being easily dispersed by wind currents. Pady (1957, p. 351) reported that fungal spores were present in the air throughout the year, but were seasonal in their distribu- t10171. with peaks in July and August, and occurred in low concentration during the winter. The intensity of the wind was also directly related to its Spore-load. He stated that additional work is necessary to determine SPOI‘e-loads at different intervals during the day. This work should be Concerned mainly with variations in temperature and humidity and their effects on the number and kinds of air-borne fungal spores. Gislen (1948, pp. 124-125), in the summary of his work, States in part: Small organisms have considerable possibilities of distri- bution by convection air currents and winds at moderate alti- tudes. Examples are given of such distribution over great distances. But as the animals are often strictly specialized ecologically (herbivores, parasites. etc.) they have particular difficulties to overcome in their new surroundings. Numbers of micro-organisms are constantly being driven up into the air to return again to earth in rain showers or downward air currents. Micro-organisms are very resistant to unfavorable factors met with in the air-sea. Some may be distributed through the air in an anabiotic stage. Being often hermaphrodite or parthenogenetic, many of them can give rise to progeny from a single individual which happens to arrive in suitable surround- ings. Their resistance to low temperature, low barometric pressure and drought is superior to that of all other organisms. Nevertheless, in comparison to larger forms,*they are very sensitive to radiations, especially ultra-violet, which seem to check their distribution more than that of larger forms. Meier (1933, p. 380) adds, as far as some green algae are cone erned, upon a 6-minute to 18-hour exposure to ultra-violet that: In the regions of ultra-violet beyond 3022 A. the approxi- mate limit of ultra-violet irradiation in nature. the green algal cells were killed. oooooooooooooooooooooooooooooooooooooooooooo Wave lengths longer than 3022 A., that is. wave lengths of 3130, 3341, and 3650 A., had no appreciable lethal effect on the algae. Gislen (1948, p. 125) asserts that: No geographical borders or barriers exist for microforms. They are often cosmopolitan, or else regionally distributed around the whole globe in certain climatic belts. Under favorable conditions. especially in humid air, the harmful influence of radiation is diminished, and microforms may be transported alive by winds over greater distances than in clear and dry weather. However. Hyman (1940, p. 71) makes the following statement: The cysts of Protozoa occur attached to grasses. and other objects. in the soil, etc.. and may be disseminated by Various agents but do not float about in the air to any extent. In Puschkarew's experiments, air inoculation of sterile cultures resulted in only 13 species, chiefly small amoebas and flagel- lates and one ciliate (Colpoda). Frequently. sterile cultures exposed to the air fail to de- ve lope any Protozoa. The ability of Protozoa to encyst and survive the effects of drying for long periods of time has been discussed by many (Hyman. 1942, p. 71; Gislen, 1940, p. 21; Kudo, 1946, pp. 147-149; Pennak. 1953, p. 15; and Galbraith and Taylor, 1950, p. 938). The formation Of a spore or cyst which can withstand desiccation favors but does not guarantee a wide distribution of particular species. As will be Seen later. my results upon exposing sterile pondwater to the air for various periods of time were very similar to those of Puschkarew. Talling (1951, pp. 160-161) states: Dispersal of small viable resting stages in wind-borne dust is frequently postulated but difficult to detect (c. f. Gislen, 1943). The exposed and drying mud on the margins of ponds would readily contribute to such aerial dust, as several authors have pointed out (c. f. Pettersson. 1940). An empirical approach to the problem is possible from observations of the entry of Small aquatic organisms into sterilized cultures or infusions left eXposed to the air. Its frequent rapidity led several algological Workers (e. g. Eddy, 1925; Pettersson. 1940; Messikommer. 1943) to emphasise [sic] the importance of wind dispersal for fI‘esh—water algae. HER/ever. the total number of species ob- tained was small, as in the earlier experiments of Puschkarew (1913) on Protozoa. in terrestrial habitats such as soil. It is Probably more appropriate to conclude. as Puschkarew did, that Such culture experiments have not established the importance of Wind in the dispersal of the aquatic micro-fauna and flora. In general. although many individual examples of the dispersal of Small aquatic organisms by wind have been established, the Over-all significance of such dispersal is still not clear. rfi Floodingand rain run—off.--The spread of algal species is aided also by flooding and rain run-off. Poretskii (1926, p. 798) "Normal phytoplankton organisms of the Nevka introduced states: by the flood into the pond rapidly decreased and had entirely disap- peared in 40 days." This would indicate that although new organisms are brought into an area by flooding. it still does not guarantee their e stablishment. Water currents and floating objects.--Water currents and floating objects also carry algae from one location to another in a given body of water but are relatively unimportant in populating a separate body of water. Biological agencies Birds.--Ridley (1930, p. 489), Taylor (1954, pp. 569-572), Savile (1956, p. 441), and others have shown that birds can transport the seeds of plants, externally or internally. relatively great distances in a viable condition. In fact it has been pointed out by Krefting and Roe (1949, pp. 271-286) and Ridley (1930. p. 489) that seeds passing filrough the digestive tract of a bird may be in better condition to germinate than if they had not been eaten. Insects.--Probably the greatest proponent of algal dispersal by insects is W. Migula (1888). In studying the scrapings from dif- ferent body parts of water beetles, Migula (1888, p. 516) recorded species of the following genera: Anabaena, Cfiharacium. Synedra. __ Oscillatoria. Scenedesmus. Navicula. Protococcus, Cosmocladium. Aphanochaete, Chlamydomonas. Cocconies. Palmella(?). Penium. Shroococcus, Hapalosiphon. Fragilaria. Encyonema, and Meridion. He concludes that the role of aquatic insects in the dispersal of ailgae is more important than either that of water birds or the air Currents. Kew (1893, pp. 62-63. 67) states that aquatic insects may also play a role in the dispersal of mollusks and fish: . John Curtis, the distinguished entomologist, expressed the opinion that the larger aquatic insects-especially the Cytiscides-might without doubt be the means of conveying fish- spawn from one piece of water to another and Mr. Wallace in like manner. discussing the means of dispersal of fishes. ob- serves that water-beetles flying from one pond to another "may occasionally carry eggs. " Irénée-Marie (1938, pp. 32, 35) reported finding members of the genus Closterium in the claws of a large Dytisid. He listed d‘Eisrmds found on the body of a dragonfly (Libellula sp.) and a beetle- Messikommer (1943, pp. 315-316) also credited dragonflies with dis- peli‘Sing micro-organisms from one body of water to another in encysted forms and spores, also as vegetative cells, if the distance was not too great. It may well be that sterile culture media exposed to the air may be contaminated by micro-organisms carried by insects rather than from the air currents. Yet in the literature Beger (1927, p. 393) and others attributed organisms found in these exposed sterile media as being air-borne and probably in the encysted stage. A study is being conducted at Cornell University, New York. by Mr. Bassett Maguire. Jr.. to advance our knowledge of the role 0f aquatic insects. especially Diptera. in the dispersal of micro- Organisms. Fish.--Dispersal of higher aquatic plants by fish has been disCussed by Ridley (1930, pp. 516-518); the role Of fish in The dis- persal of algae has been mentioned by Irénée-Marie (1938. p. 31). Lef‘evre (1940, pp. 347-349), Velasquez (1939. pp. 386, 389. 403). Tiffany (1927, p. 303), and others. The algae can be carried within the digestive tract and then Clefecated in a viable condition when the fish reaches a new location. or carried externally on the body. especially. along the edge of the scales. Tiffany (1927, p. 33) remarked in respect to the species and varieties of algae found in an identifiable condition in the digestive tract of a young gizzard shad: The species and varieties of algae identified from this young gizzard shad numbered 57, distributed in the following groups: 11 Myxophyceae, 3 Euglenidae. 1 Phaeophyceae [Dinobryon setularia], 2 Heterokontae, 13 Bacillariae, and 27 Chlorophyceaef fl Velasquez (1939, p. 403) removed the contents from various Sections of the digestive tract of the gizzard shad (Dorosoma cepedi— EEELQ). cultured these under sterile conditions. and found: The species and varieties of viable algae are practically the same throughout the alimentary canal. It may therefore be concluded that whatever species are quickly destroyed and di- gested are quite immediately effected near the mouth end of the alimentary canal. In this connection, the large number of com- mon genera that were not recovered at all in culture is signifi- cant. No specimens or only an insignificant number even turned up in culture of any of the following groups always present in ordinary freshwater habitats: (a) Bacillarie, (b) Volvocaceae group, (c) Dinobryon of the Heterokontae. and (d) filamentous green algae. . I There survived 30 species and varieties of Chlorophyceae: 12 species and varieties of Myxophyceae; 4 species of Bacillariae. 2- species of Heterokontae; and 1 species of Euglenophyceae. The order Chlorococcales (Chlorophyceae) had the greatest number of viable algae. It seemed that cell wall modifications or rather special secretions resisted the digestive fluids of the fish. Faecal material from three fresh-water fish (Syprinus Earpio. Mus rustilus‘. and Erama brama) was cultured by L‘efevre (1940, 99° 733-739) and the most common algal groups were: Protococcales. Flagellates. Cyonophyceae. Dinoflagellates. and Volvocales. The 10 desmids and many diatoms seem particularly susceptible to destruc- tion by intestinal secretions. Irénée-Marie (1938, p. 31) states that fish can carry count- less desmids cemented to their scales which may then become de- tac bed by the fish brushing aquatic vegetation. Reptiles.--Most of the work on the role of reptiles in the dispersal of algae has been carried out on turtles (Edgren. Edgren. and Tiffany, 1953, pp. 733-739). Ridley (1930, p. 515) does mention the role that lizards and tortoises play in the dispersal of higher Plants but does not discuss the algae. Vinyard (1953. PP- 63-64) mentioned that: . . Certain aquatic or semi-aquatic animals are excep- tionally good algal habitats in themselves. The dearth of in- formation on the species of algae occurring on such substrates. as well as on the identity of such animals bearing algal growths. has made it apparent that much useful information might be ob- tained on the nature of these plant-animal relationships. In his study of algal growths on some turtles in Oklahoma he 118led the species of algae found on the various turtle species. Neill and Allen (1954, p. 583) stated that ". . as a turtle moves from Pond to pond, it may disseminate the epicolous alga." Painted turtles (Chrysemys pigta) have been observed by the all’thor near the W. K. Kellogg Gull Lake Biological Station of Michi- gan State University traveling overland between ponds carrying an ll obvious algal growth upon their backs as well as various species of leeches adhering to the edge of their carapaces. The turtles prob- ably are also important in the dispersal of algae from one body of water to another. Amphibians.--Ire’née-Marie (1938, p. 32) has considered the possibility of frogs dispersing algae. Of the twenty-five frogs ex- amined. only one did not carry algae externally. From the body of One small frog he obtained 326 desmids representing nine different genera. Migula (1888, p. 517) asserts that water beetles and frogs Play an influential part in the dispersal of algae and that frogs may Carry more kinds of microscopic plants and animals than do water beetles. Salamanders may also play a role in dispersing micro- Organisms. Mammals.--Irénée-Marie (1938. pp. 31-32) trapped a mink (MBEEM vision, probably) and removed from its fur 169 desmids. The Slxteen species in nine genera, and some unclassified cells. genus Closterium was especially abundant. Therefore it seems probable that mammals may contribute to the transport of algae and protozoa as well as fish eggs. snail eggs, e1 Cetera, from one body of water to another. but there is no direct evidence to support this assumption. 12 Man.--Talling (1951, p. 160) states: Man himself has been an agent in the disPersal of fresh water organisms. In addition to accidental transportations, the vexed question of artificial tranSplantations belongs here. . . . Also, even the energetic field naturalist may be unconsciously responsible for extending the range of a species, as he empties the residue of his collections of the day into some convenient pond or stream. Man has had a great influence upon the distribution of various species of higher plants and animals through modification of the en- vironment and introduction of new species. Probably a long time must elapse before we shall have measured the total effect man exerts 0n the environment. Although we do know that the introduc- tion of even one species of higher plant or animal into a new area can have great effects upon the ecology of that area. the effects of a “GWIy introduced protozoan or algal species upon a given plankton population has not as yet been studied. Eddy (1925, p. 143) states: . In all probability, unicellular algae, such as Diatoms and Euglena, are pioneers of the initial stage. Seeding condi- tions for these forms are generally much better than for the filamentous types. From the early appearance of Flagellates and Diatoms in sterile cultures and initial stages, it is very eVident that the encysted forms of these species are very widely and readily dispersed. The higher types of filamentous algae are either not so readily dispersed, or do not possess so wide a r ange of adaptability and require more favorable conditions than Offered by the initial stages. 13 As suggested by Dr. K. L. Osterud (1955), the study of the initial bacterial. protozoan. or algal invaders of a pond and their modification of the aquatic environment would be well worth investi- gating. For an additional review of the various problems in the dis- persal of micro-organisms. The Elements of Cfihanfice in Pgnd E93- lations. by J. F. Talling (1951). is suggested. CHAPTER II HISTORY OF THE PROBLEM An exhaustive search of the literature reveals only a few references to the problem of dispersal of micro-organisms by birds. Charles Darwin was probably the first investigator to conduct any type of controlled experiment to demonstrate that waterfowl may be important in the transport of aquatic organisms. Darwin (1859. pp. 302, 304) states: When ducks suddenly emerge from a pond covered with duck-weed. I have twice seen these little plants adhering to their backs; and it has happened to me. in removing a little duck-weed from one aquarium to another. that I have uninten- tionally stocked the one with fresh-water shells from the other. But another agency is perhaps more effectual; I suspended the feet of a duck in an aquarium. where many ova of fresh-water Shells were hatching; and I found that numbers of the extremely minute and just-hatched shells crawled on the feet. clung to them so firmly that when taken out of the water they could not be jarred off. though at a somewhat more advanced age they Would voluntarily drop off. These just-hatched molluscs [sic]. though aquatic in their nature. survived on the duck's feetT-i-n damp air, from twelve to twenty-hours; and in this length of time a duck or heron might fly at least six or seven hundred miles. and if blown across the sea to an oceanic island, or to any other distant point. would be sure to alight on a pool or rivulet. We should not forget the probability of many fresh-water forms having formerly ranged continuously over immense areas. and then having become extinct at intermediate points. But the 14 15 wide distribution of fresh-water plants and of the lower animals, whether retaining the same identical form or in some degree modified, apparently depends in the main part on the wide dis- persal of their seeds and eggs by animals, more especially by fresh—water birds, which have great powers of flight, and nat- urally travel from one piece of water to another.. According to Talling (1951, p. 159), Darwin "performed the simple experiment of dipping a severed duck's foot in an aquarium to test the viabilities of aquatic organisms left stranded on the feet; a repetition of this experiment in nature would be of interest." Following Charles Darwin, the first person to do actual re— search to discover whether algae may be carried externally on water- fowl was Jules de Guerne. His work, Sur les dissemination des organismes d'eau douce gar les Palmip‘edes, published in 1888, is certainly the first major contribution to the study of waterfowl in the dispersal of micro-organisms. Although he conducted the original research, he is seldom quoted directly in the literature, and then only briefly. Reference had been made to De Guerne's work by Zacharias (1888, p. 369), Huber-Pestalozzi (1938, p. 72), and a few others, but most English-writing ecologists have preferred to quote German workers (i.e., Zacharias, 1888; Zschokke, 1900; and Beger, 1927). 15 wide distribution of fresh-water plants and of the lower animals, whether retaining the same identical form or in some degree modified, apparently depends in the main part on the wide dis- persal of their seeds and eggs by animals, more especially by fresh-water birds, which have great powers of flight, and nat- urally travel from one piece of water to another.. According to Talling (1951, p. 159). Darwin "performed the simple experiment of dipping a severed duck's foot in an aquarium to test the viabilities of aquatic organisms left stranded on the feet; a repetition of this experiment in nature would be of interest." Following Charles Darwin, the first person to do actual re- search to discover whether algae may be carried externally on water- fowl was Jules de Guerne. His work, Sur les dissemination des organismes d'eau douce par les Palmipledes, published in 1888, is certainly the first major contribution to the study of waterfowl in the dispersal of micro-organisms. Although he conducted the original research, he is seldom quoted directly in the literature, and then only briefly. Reference had been made to De Guerne's work by Zacharias (1888, p. 369), Huber-Pestalozzi (1938, p. 72), and a few others, but most English-writing ecologists have preferred to quote German workers (i.e., Zacharias, 1888; Zschokke, 1900; and Beger, 1927). 16 Because De Guerne's work has a direct bearing on this re- search, I have included my translation of the original French publi- cation in the Appendix. Thus, in tracing the history of the problem, one may regress from Smith (1933) to Beger (1927) to Zacharias (1888) to De Guerne (1888) and finally to Darwin (1859). Between 1850 and 1875, Sir Charles Lyell, A. R. Wallace, Charles Darwin, and their associates probably exchanged ideas freely. This group undoubtedly should re- ceive the credit for having laid the foundation for much of our pres- ent knowledge of diSpersal and distribution of plants and animals. Kew (1893) in The Dispersal of Shells gave many indications as to how various mollusks may be dispersed throughout the world. He is probably most frequently quoted by English ecologists. Various workers have reported finding snail eggs, pieces of higher aquatic plants, mollusks, insects, et cetera, adhering to the feathers, feet, and bills of waterfowl. Darwin (1859) observed Lemna adhering to the feathers. and young fresh-water snails ad- hering to the feet of ducks. Later, in 1878, he reported a clam (Unio complenatus) attached to the toe of a blue-winged teal (Querquedula discors) shot in Massachusetts. Zacharias (1888, p. 368) stated that Humbert found winter eggs of crustacea on the feathers of wild ducks, and Kew (1893, pp. 47, 52) reported that a l7 mallard was shot "in the Sahara desert a hundred miles from water" with mollusk eggs "attached by the glutinous coating to one of the feet." He also stated that pieces of aquatic plants may adhere to the body of waterfowl when they leave a body of water. Reid (1892, p. 278) stated that the nest of the stickleback fish is attached to water plants and would likely be tranSported with the plants when they become attached to waterfowl. . Molluscs might possibly be carried in the crops of birds considerable distances, and others be distributed and es- tablished in new districts or on islands, as the living shells might be ejected from the cr0ps, or the birds might be killed by birds of prey [or hunters] and the contents of the stomach dislodged and scattered. [Kew, 1893, p. 161] I picked a living bark beetle out of the feathers of an owl knocked down in flight in the highlands of Fiji. Owls have been seen at sea 1,000 miles from the nearest land. [Zimmerman, 1948, p. 54] Saville (1956, p. 441) stated that Lemna and Spirodela were probably spread largely by adhering to the body feathers and feet of waterfowl as they rise from ponds in which these plants are growing. There is no wish on the part of the author to imply that these early reports are untrue. But a more detailed account of these observations discussed in the literature would eliminate much misinterpretation in some ecological reports. Oneis inclined to be critical of another's work but sometimes it is more difficult to pro— duce something better. 18 Also the author had observed Lemna minor and filamentous green algae adhering to the feathers and feet of ducks when they were first removed from the water. After the ducks had been hanging in the air 10 minutes, the Lemna minor and algae were not found (Chapter VI). We will do well to keep in mind Darwin's remark, "How ignorant we are with respect to the many curious means of occasional tranSport.” [Zimmerman, 1948, p. 62] Ster (1924, p. 141) asserted that the flights and migrations of wading and swimming birds are of great importance in the distri— bution of organisms, perhaps eSpecially for local habitats, and that much research remains to be done concerning the possibilities and means of the distribution of algae. Some of this much-needed work, done recently by Irénée- Marie (1938) and Messikommer (1948) has aided us greatly in ob- taining a clearer idea of the role which waterfowl play in the trans- port of algae. In Flor Desmidiale de la Region de Montreal (1938, pp. 32, 33) Fr. Irénée- Marie described the washing of the feet of a Blue Heron in filtered water after observing its flight between peat bogs. The following desmids were recorded: 19 Genus Number Observed Vlklumber of Species Closterium 31 3 Penium 5 . 1 Rleurotaenium 5 2 Triplvocervas‘ 32 2 Sitaufirastrum‘ 1 8 3 Spongylosiuin 2 filaments. 17 cells 1 Netrium 9 1 Cosmarium 9 2 A total of 126 cells of 15 species representing eight different genera were found. Iréne’e-Marie had declared a belief that many specimens remained on the walls of the vessel containing the wash- ings. He also killed ducks before they could land in the water of the peat bog. The washings of the plumage of one duck yielded 517 desmids representing 31 species and 13 genera. In _A__l_g_e:1nachweis in Entenexkrementven (1948. pp. 23-24) Messikommer reported his findings from the direct microscopic ex- amination of fresh faecal material from ducks. He found fragments of higher plants, empty diatom frustules of gynedra. Soccengis. Fragilaria. Cymbella. Epithemia. Navicula. and gqrnphonema. an 20 unclassified living ciliate. epidermal cells of sedges. fragments of different filamentous green algae consisting of two species of Oedogonium and Microsporia quadrata and often Tribpnema vulgar}: Staurastrum cingullim. Scenedesmus ecornis‘, Spirogyra sp.. moss leaves. vessels of higher plants. and pieces of insects also were observed. Klingle (1940. p. 191) used sterile water to wash the feet of sixteen Spotted Sandpipers (Actitus magnlaria) which he shot on Inaqua Island in the West Indies. He found eleven small seeds. two species of desmids. microscopic green algae. and a number of un- classified amoeba-like organisms. There may have been others like Klingle who have washed birds in the field making brief comments about their observations. CHAPTER III HUNTERS' DATA SHEETS AND FIELD DATA A preliminary investigation concerning the problem of algal dispersal by waterfowl was begun in the fall of 1953. The objectives of tlus initial investigation were to determine (1) what micro- organisms are carried externally on waterfowl. and (2) in what stage they are carried; i.e.. vegetative. encysted, or in a spore Stage. By use of the hungers' data sheet (Appendix A) which was a form that the hunters were requested to fill out. the following infor- mation was secured: (1) name of the bird shot; (2) sex of the bird; (3) Position of the bird when shot; i.e.. coming into. leaving. or Swimming in the body of water; (4) name and location of the body of Water; (5) date; (6) time of day the bird was shot; (7) name and address of the hunter; and (8) additional remarks. The heads and feet of the ducks were removed by the hunters and placed in a new number 5 paper bag along with the correspond- ing hunters' data sheet. Later washings were made from the bills and feet using boiled pondwater. 21 22 Methods The glassware used in this research. after being cleaned, was boiled in tap-water for 30 minutes and allowed to dry on fresh Paper toweling. Distilled water was boiled 20 to 30 minutes in 250-ml. Erlenmeyer flasks. capped with cotton plugs. and allowed to 0°01 to room temperature. The sterile water was used for the Sterile washings of the birds' feet and bills. The medium for culturing algae and protozoa consisted of a Cla3"‘1c>am soil moderately rich in humus which was obtained from a univeI‘sity farm (Southworth). The soil was boiled in distilled water for 30 minutes. allowed to stand for periods of time varying from three hours to two days. and again boiled for 30 minutes. It was then poured into sterile fingerbowls. covered with sterilized glass plates . and allowed to cool. Later the medium was inoculated with centrifuged material from the bird washings. With each preparation 0f Inedium there were at, least two controls to which no foreign matter was added. The feet and bills of the birds were washed by swirling in the boiled distilled water for three minutes. although some speci- m . . ens were allowed to soak overnight. On a few occasmns. the w . . ash-lug periods were even longer. These washings were made 23 from one hour to as long as three months after the shooting of the bird. The birds had been stored in the Zoology Department cooler at approximately 40 degrees Fahrenheit. Approximately one-third 0f the washing was centrifuged and studied immediately after wash- ing, another third was placed in culture media. and the remaining third was preserved in lO-percent formalin solution. Samples of four to twelve drops of medium were removed from the surface. the middle. and the bottom of each fingerbowl. These samples. examined microscopically. were considered to con- tain representatives of the algal and protozoan inhabitants in the Cultur e. Field Collections Ete‘rs' data sheet The waterfowl were collected from various hunters in differ— ent localities. Six common snipe (Capella gallinago) were Shot by Dr. A. D. Geis at about 9:00 a.m. on October 15. 1953. They were flushed from the mud flats along the edge of Crooked Lake (Barry County. Michigan). The snipe were kept at air tem- perature until 3:00 p.m. Then they were placed in the Zoology De- ' partlruant's cooler at 40 degrees Fahrenheit. They remained in the 24 cooler until 4:30 p.m., October 16. 1953; then washings were made of their feet and bills. The feet of three snipe were washed for approximately two minutes in boiled distilled water. A sterilized scalpel was placed between the toes of the birds and shaken vigorously for several sec- OndS to remove any material which might have lodged there. A second beaker was used to wash the feet of the remaining three birds. The bills of four birds were washed in a third beaker. After being centrifuged. approximately the upper three-fourths of the liquid was decanted. In Table I are listed the other birds that were collected. The organisms observed microscopically are recorded in Table 11. m. Washingon On April 25, 1955. at 9:30 a.m. a Mallard duck (£2913. EIaEZrhynchos) was taken from Lake Washington at Seward Park. The Excess water was shaken from its feet. After the bird had been held in the air for five minutes. each foot was placed in a Small jar containing boiled pondwater and shaken vigorously. The was‘l‘la‘xigs were centrifuged at low speed and examined at 11:30 a.m. A filament of Oscillatoria sp.. a few unclassified small (3,“) green unicells. debris. and pieces of diatom frustules were observed. TABLE I BIRDS EXAMINED IN THE HUNTERS' DATA SHEET STUDY, FALL, 1953 25 Period Quan- of Time Bird tity of Body of between . Name . No. Birds Water Shooting Shot and Washings 1 l Ruddy Duck (Oxyura Lake 4 days jamaicensis) Lansing 2 2 Wood Duck (Aix sponsa) Rose Lake 5 days 3 1 Black Duck (Anas Rose Lake 8 days rubripes) 4 1 Redhead (Ay‘thya Lake 2 hours americana) Lansing 5 2 Coot (Fulica americana) Lake 2 hours Lansing 6 1 Canada Goose (Branta Wintergreen 3 days canadensis) Lake 7 1 Blue Goose (Chen Saginaw Bay 22 days caerulescens) Marshes 8 1 Green-winged Teal Saginaw Bay 22 days (Anas carolinensis) Marshes 9 3 Black Duck (Anas Saginaw Bay 22 days rubri pes) Marshe s 10 6 Black Duck (Anas Macks Creek. 13-2-4 rubripes) Stanwood days TABLE I (Continued) 26 Period Quan- of Time Bird tity of Body of between . Name . No. Birds Water Shooting Shot and Washings ll 2 Buffle-Head Duck Lake 12 hours (Bucephala albeola) Lansing 12 1 Ruddy Duck (Oxyura Lake 12 hours Elmaicensis) Lansing l3 3 Coot (Fulica americana) Lake 12 hours Lansing 14 1 Canada Goose (Branta Wintergreen 26 hours canadensis) Lake 15 6 Common Snipe (Capella Crooked 31 hours gallinago) Lake 27 TABLE II ORGANISMS OBSERVED ON MICROSCOPIC EXAMINATION OF WASHINGS FROM THE BIRDS IN THE HUNTERS' DATA SHEET STUDY, FALL, 1953 Portion of B1315? Bird Zrefizf::::? Organisms Observed Washed 1 Bill, Feet Cent. Wash.a No living algae or protozoa 2 Bill, Feet Cent. Wash. No living algae or protozoa 3 Bill, Feet Cent. Wash. No living algae or protozoa 4 Feet Cent. Wash. _E_u_g1ena gracilis, fungal Spores Cult. Mediab Nostoc verrucosum, Oscillatoria limnetica 5 Bill, Feet Cent. Wash. Oscillatoria amphibia Cult. Media Oscillatoria amphibia 6 Feet Cent. Wash. Navicula sp., unclassified ciliates 7 Bill, Feet Cent. Wash. Colpidium sp., fungal Spores 8 Bill, Feet Cent. Wash. No living algae or protozoa 9 Feet Cent. Wash. No living algae or protozoa Cult. Media No living algae or protozoa Bill Cent. Wash. Geminella minor Cult. Media Geminella minor aCent rifuged Washing. bCulture Media. 28 TABLE II (Continued) Bi rd No . Portion of Bi rd Washed Preparation of Material Organisms Observed 10 ll 12 13 14 Feet Bill Feet Bill Feet Feet Bill Feet Cent. Cent. Cent. Cult . Cent. Cent. Cult. Cent. Cult. Cent. Cent. Wash. Wash. Wash. Media Wash. Wash. Media Wash. Media Wash. Wash . Peranema sp., Navicula sp., Kirchneriella subsolitaria Navicula sp., Peranema sp., Collembola Scenedesmus abundans, S. ar- cuatus, Gloeocystis gigas, Pinnularia sp. Gloeocystis vesiculosa, G. gigas, Aphanocapsa elachista, Oscilla- toria Agardhii Oscillatoria limnetica, Gloeo- cystis sp., Gloeocapsa aertigi: nosa Oscillatoria limnetica, O. Agard- h_i_i_, Scenedesmus arcuatus Oscillatoria Sp. Scenedesmus arcuatus, Gloeo- cystis vesiculosa, Sphaerella lacustris Nostoc sp., Oscillatoria sp. Pediastrum Boryanum, Aphano- capsa elachista, Oscillatoria amphibia Gloeocystis vesiculosa, Palmella mucosa 29 TABLE II (Continued) Portion of B' d ' 1r Bird Preparation Organisms Observed No. of Material Washed 14 Feet Cult. Media Colpoda steini Bill Cent. Wash. Sphaerocystis Schroeteri Cult. Media Protococcus viridis, Trochiscia granulata, Sphaerocystis Schroeteri 15 Feet Cent. Wash. Frontonia sp., two unclassified and amoebae, Navicula sp., Phacus Cult. Media spp., Geminella sp. Bill Cult. Media Nostoc sp. 30 The washings were placed in the Zoology Department cooler until May 2. Upon examination of the washings May 2. the follow- ing were seen: Ulothrix sp.. Oscillatoria sp.. Monas sp.. and w—fffi Chilomonas sp. On May 11 Ulothrix sp.. Oscillatoria sp.. Monas fvw— Sp., an amoeba. fungal spores. and unclassified cysts were present. The culture was found to contain the same organisms on May 16 with the addition of Euglena Sp. and an unclassified holotrich. One-half of the April 25 washings were added to a sterile hay infusion and the other half to a wheat culture medium on May 2. On May 19 the hay infusion contained Xorticella sp.. Monas sp.. Colpoda sp., bacteria. and a Hartmonella-like amoeba. Colpoda sp., Paramecium bursaria. Vorticella sp.. and unclassified cysts were fiv m ,_ present on May 27 and 28. The wheat culture contained Vorticella sp.. Monas sp.. Ulothrix sp.. and Holotrich: Pithothorax Sp. On May 30 the wheat culture contained Colpoda Sp.. Ulothrix sp.. Oscillatoria sp., Bodo Sp., Monas Sp.. and Plagipcampa sp.. and the hay infusion contained only bacteria and Paramecium bursaria. Pine River . Michigan. On November 7. 1955. at 10:30 a.m.. a Goldeneye Duck (Glaucinetta clanggla americana) was flushed from the main stream of the Pine River between St. Louis and Porter. Michigan. The 31 bird was in the air an estimated 30 to 45 seconds before being shot down; it fell into the grass on a high bank (8 to 10 feet) about 20 feet from the river. The feet and bill were washed immediately by being shaken vigorously for two minutes in vials of sterile pondwater.- At 3:50 p.m. of the same day the washings were returned to Michigan State University and planted in culture flasks of soil-water media. Examination of the material on November 26 and December 9. 1955, showed the following organisms: Feet washings: Gyrosigria Sp., Gomphonema sp., Navicula Sp., Cyclotella Sp., Gloeocystis sp., ‘Nannochloris bacillaris, Chlamydomonas Cienknowskii, Scenedesmus longgs, S. armatus, Ulothrix sp., and CladOphora Sp. Bill washings: Ulothrix Sp., Euglena Sp., Gomphonema sp., and Oscillatoria Sp. A Gadwall Duck (Anas strepera) was flushed from an arm of the Pine River at 11:00 a.m. Duckweed (Lemna minor) was abundant in the area, although the duck may not have been in it. The bird was in the air 30 to 45 seconds before being shot. It fell in the grass about 50 yards from the river. Washings of the feet were made iInmediately. These washings were planted in culture flasks at 3:50 p.m. The feet appeared perfectly clean. Even under the toe nails, no dirt particles were visible. Examination of the culture showed Nannochloris bacillaris, Scenedesmus Spp., Scenedesmus fi—f 32 abundans, glothrix sp.. Mougeotia sp.. Spirogra Sp., Cyclotella sp., Navicula sp., Gomphonema Sp.. Gyrosigzna Sp., and Oscillatoria Sp. Samples were also planted from the upper portion of the gul- let of each bird. The gullet of the Goldeneye contained two seeds of Potomogeton Sp. which germinated. and a filament of Spirogyra sp. Also present were Gloeocystis sp.. Ankistrodesmus convolutus. Mougeotia sp.. Gyrosigma sp.. Navicula sp.. Cyclotella sp.. and protozoan: Monas-like flagellates. The gullet of the Gadwall showed Euglena sp.. Navicula sp.. Gyrosiglna sp.. Spirogra sp.. Lepocinclis sp.. Chromulina sp.. Nannochloris sp.. and rotifer: Bdelloidea. fi‘r—v St. Joseph River. Michifln A Mallard drake (Arias platryllyrnphos) was shot by Dr. M. D. Pirnie on November 13. 1955. in the St. Joseph River rice beds. Excess water was shaken from the duck and it was placed in a clean plastic bag for protection from contamination. The bird was kept in the Zoology Department cooler until November 16; then the feet and the bill were washed and the cultures made. On December 3 the following forms were noted in the culture WT. media. Feet washings: Scenedesmus spp.. S. quadricauda. Nan:- nochloris bacillaris. Ankistrodesmus spp.. Shlorella vulgaris. Syn. edra sp.. Cyclotella sp.. and Navicula sp. On December 9 33 Kirchneriella sp. was found in small numbers in addition to the above organisms. Bill washings: Navicula sp.. Valkamphia-like amoebae, and two species of unclassified flagellates. At this time there were large numbers of protozoa but no green or blue-green algae. Port Sanilac . Michigan On July 13. 1957. a Spotted Sandpiper (Actitis macularia) was Shot while walking in moist sand and debris on the shore of Lake Huron approximately one and one-half miles south of Port Sanilac. Michigan. The feet were washed immediately by being placed and shaken in a vial of boiled well-water for three minutes. A saturated solution of mercuric chloride was added to the vial in sufficient quantity to double the original volume of well-water. This solution remained standing for three minutes to allow organisms to settle out and then the upper half of the solution was decanted. An equal vol- ume of 6-3-1 (Transeau's) solution was added to the remaining por- tion containing the organisms. A microscopic examination on August 1. 1957. showed the following organisms to be present: Quadricula Closterioides, Pedi- astrum Boryanum. filaments of Rhizoclonium sp. with Characium sp. attached. Phormidium sp.. Synedra Sp.. Frag} aria sp. Tabellaria- sp. 34 frustule. pieces of a diatom frustule (Amphora sp.). and epidermal hairs of higher plants. Another Spotted Sandpiper was shot on dry wood chips 20 yards from the shore of Lake Huron. Upon microscopic examination only pieces of insect exoskeleton and an Alternaria-like fungal spore were found. in the washings. An Eastern Belted Kingfisher (Meggceryle alcygn) was shot out of the air; it fell into moist sand a few feet from the lake. Fragilaria sp. frustules. Navicula sp.. Synedra sp. frustule. pieces of insect exoskeleton. epidermal cells of higher plants. a few un- classified cysts or spores. and debris were found in the washings. A Purple Martin (P303118 Ems subis) shot under the same conditions yielded only debris and a few unclassified spherical cells (Cyanophyta) 2’“ in size upon microscopic examination. On~July 15 two Ring-billed Gulls (Larus delawarensis) were shot. One fell in dry grass on a high bank about 30 feet above the level of the lake. Its feet were washed in a vial of boiled well— water. A microscopic examination on August 1 revealed one cell of Gloeogystis gl'gas. a broken Ostracod valve. pieces of insect an- tennae. unclassified spherical cysts or spores. 3/u in diameter. and debris . 35 In the culture of the bill washings of the gulls. bacteria. an unclassified fungal mycelium. and cysts or spores were observed. About 2 ml. of the liquid contents of the upper part of the gullet were also cultured. Peranema-like flagellates and a brown fungal spore were seen. About 3 ml. of faecal material were obtained from the bird and cultured. This culture on August 1 Showed a Navicula sp. frustule. pieces of insect. bacteria. and debris. The second gull fell in dry sand on the‘ shore. Four to six drops of the settled washings revealed only sand grains. hairs. bacteria. pieces of insect antennae. and debris. The gullets were packed with earthworms. indicating that these birds had not been in the water for some time prior to the shooting. Gulls are frequently seen feeding in farm fields one-half mile from the lake shore. 1" a1.‘ ...¢. CHAPTER IV CONTROLLED EXPERIMENT. 1955 Controlled experiments were devised to demonstrate and evaluate the role of waterfowl in the dispersal of algae. This method has many advantages over the interpretation of data gathered from waterfowl shot in the field by hunters. Important factors not available from the Hunters' Data Sheet were: (1) where the bird was just prior to being shot. (2) how long the bird had been in the air before being shot. (3) where the duck fell after being shot. and (4) what microscopic organisms occur in these environments. Description of Wintergreen Lake The controlled experiments were conducted on Wintergreen Lake at the W. K. Kellogg Bird Sanctuary in section 8. Ross Town- ship. Kalamazoo County. Michigan (Figure 1). Fetteroff (1952. pp. 4, 7) stated that Wintergreen Lake has an area of 39.33 acres. has a mean depth of 7.56 feet. and a volume of 297.16 acre feet. the maximum depth being 21.33 feet. Although 36 37 334 somewaowsfls mo 33> 3?ko 3356952, .9 .m we $3.509 .2 manna 38 there are no permanent feeder streams. the lake has a drainage area of 530 acres. He continued: At the south end of the adjoining swamp area. there is an outlet which empties into Gull Lake. a half mile distant. Pre- sumably. springs located on the north and northeast Shore keep the lake at a fairly constant level. Wintergreen Lake lies in the Kalamazoo-Mississianawa morainic system outwash plain. This plain is characterized by numerous lakes in the morainic basins and in the pits in the outwash plain. Wintergreen Lake is one of many small pit lakes in the vicinity. Bottom deposits are variable. The south and west shore is [generally] pulpy peat to a depth of three feet. where marl be- comes intermixed with it. Marl is predominant to a depth of about twelve feet in all other parts of the lake except the east and northeast shore. These shores are exposed to wave and wind action and are sandy to a depth of 2.5 feet where marl again becomes predominant. Beyond the twelveefeet depth the bottom is of a fine organic ooze. The area in which the research was conducted was in shallow water on the west side of the lake. The substratum was sandy and covered with silt. Because of the activity of the ducks. there was very little silt within the water pen. Matted algae floated freely nearby during the month of July and in the first two weeks of Aug- ust. A dense bed of lily pads was growing three feet beyond the deeper end of the water pen. There was no wave action during the Period of investigation. 3 9 Materials A laboratory was available in the boathouse at Wintergreen Lake where the experiments were conducted. Carboys of tap-water and distilled water were brought in for water supply. A culture rack was placed at the northeast entrance of the boathouse to hold the flasks after inoculation. From approximately 6:30 a.m. until 9:30 a.m. each day. the flasks received direct sunlight and the re- mainder of the day. diffused light. During the three weeks in which the experiments were conducted. the air temperature in the boathouse fluctuated from 25 degrees to 30 degrees Centigrade. Glassware (slides. pipettes. and cover slips) was washed in a detergent solution. composed of one-fourth cup of Tide to nine cups ' of tap-water. They were then boiled in distilled water for one hour on two consecutive days and allowed to cool to room temperature. Plastic boot squares described below were washed with the detergent solution and rinsed well with sterile distilled water before use. Solution prepared in the same manner was also used to scrub the air cages and holding funnel before and after each experiment. The culture medium was prepared by placing 72 grams of sandy loam soil and 100 m1. of distilled water in 250-ml. Erlenmeyer 40 flasks. These were plugged with cotton. The flasks were then autoclaved at 15 to 20 lbs. pressure and‘at 100 to 120 degrees Centigrade for one hour on each of two consecutive days. Funnel traps. constructed from perma-netting over wooden frames. were used to capture the ducks. The traps were approxi- mately 12 feet long. four feet high. and five feet wide. The entrances to them were funnel-like in shape; i.e.. the inner portion of the open- ing was smaller than the outer portion (Figure 2). An enclosed pen of perma-netting was constructed in the water where the depth was 10 to 15 inches. The dimensions of the pen were: length. 14 feet; width. 5-1/2 feet; and height. 6-1/4 feet. It was used during the experiment to retain the ducks within a given en- vironment. As the study progressed. I learned that the dabbler ducks required a resting place at night. To provide this. an iron pipe two inches in diameter and three feet long was placed across one corner of the water pen about two inches below the surface of the water (Figure 3). Two cages were constructed for holding the ducks in the air. One cage was made from an orange crate. the other from an apple box two feet long. one foot wide. and one foot high. Chicken wire with two-inch mesh was placed across the bottom of the air cages to protect the ducks' feet from faecal contamination. The cages Figure 2 . Funnel Trap 41 Figure 3. Water Pen 43 were elevated 43 inches above the ground. They were placed be- tween two trees which were 30 feet apart and provided shade except between 10:00 a.m. and 3:00 p.m. (Figures 4 and 5). Another piece of apparatus constructed was a metal holding funnel with two five-inch slots cut in the underside. The sharp edges of the slots were covered with tape to prevent injury to the ducks ' legs. This funnel was used to hold the ducks and restrain their legs while work was being done on their feet (Figure 6). Clean plastic squares were cut from a clear plastic table- cloth. These were cupped. filled with 5 ml. of boiled distilled water. and were then tied securely with string around the legs of the ducks to form boots. Controls Three types of controls were used during this period. (1) Cultures were planted with lake water from the water pen. (2) Boiled pondwater from open fingerbowls was cultured after standing on the air cages. (3) One autoclaved flask of the soil- water media was kept free of inoculation and examined micro- scopically, The controlled experiment consisted of the following: 44 Figure 5. Duck in Air Cage Figure 6. Duck with Plastic Boots in Funnel 46 l. The ducks were captured in the funnel traps which had been placed in shallow water along the shore of Wintergreen Lake. Corn was used as bait to entice ducks to enter the trap. 2. According to the phase of the experiment being conducted. the ducks were washed for three to five minutes in a pan of the de- tergent solution at that time. They were then placed in the water pen. 3. While the ducks were in the water pen for varying periods of time. a check of the lakewater in the pen was made to determine the major organisms present. This check was accomplished by culturing 10 ml. of lakewater from the water pen (Table IV). and also by recording the organisms found on both the upper and lower surfaces of glass slides which had been exposed to the lakewater inside the water pen. 4. After removal from the water pen. the ducks were placed in the air cage for various lengths of time. 5. The ducks were placed in the metal holding funnel and the plastic boots were tied about their feet to obtain the desired washings. The boots were squeezed several times to impose an agitator-type motion upon the ducks' feet to wash off any organisms Present . 47 6. The washing from one foot was centrifuged at low speed for five minutes. All but 1 ml. at the bottom was decanted. The remainder was Shaken and three drops of it were removed for microscopic examination in the fresh state. The washing from the other foot was poured into flasks of autoclaved soil-water medium which were placed in the culture rack in the boathouse. 7. The pH of the culture flasks before inoculation was 6.1 as obtained with a Beckman pH meter. At the conclusion of the culture examination the pH varied from 5.5 to 6.7. Flask 31 (Table III) to which Sphagnum had been added gave a pH reading of 4.8 8. The culture flasks were kept in the Botany Department 0 greenhouse at Michigan State University between October 18. 1955, and January 25. 1956. The air temperature during this period ranged from 67 degrees to 76 degrees Fahrenheit. 9. Following completion of the culture examination. approxi- mately 5 m1. of the cultured material were added to vials containing an equal amount of 6-3-1 solution. These were preserved for a Permanent reference of the organisms cultured during the 1955-1956 period. 10. Microscopic examinations of the cultures were made three times during the eight months after inoculation. The method 48 of examination was to mix by swirling the liquid portion of the flasks and pipette out 5 m1. into a small beaker. This liquid was mixed again and three drops were withdrawn for microscopic study. Five transects were counted of each drop under high dry objective (430x) and two to five transects under low magnification. The or- ganisms observed were classified and recorded (Table III). Aerial forms observed ‘in the 1955 cultures were Anabaena sp.. Asterococcus sp.. Chlamydomonas sp.. Chlorella Sp.. Chlorococcum sp.. Gloeocystis gigas. Palmelia Sp.. Cyoococcus sp.. ffivi Gloeocapsa sp., Oscillatoria sp., protozoa: Anisonema-like flagellate. and an unclassified flagellate. fungal hyphae: Alterparia sp.. and a fern prothallus. 49 TABLE III DATA FROM CONTROLLED EXPERIMENTS WITH CLASSIFICATION OF ORGANISMS, 1955 Tide . . t : Da e Wash Time Time Boot Uncultured Flask Duck Cul- , in in , Time . Wash Washing No. No. ture , HZO Air , , , in Time Examination Plant , Pen Cage Min. 1 Aug. 4 0 3 15" 4, hrs. 1 955 15" 2 Aug. 3 0 3 30" 4, hrs. 1955 28" 3 Aug. Culture Control 4. 1955 4 Aug. 0 0 3 38" 6, hrs. 1955 TABLE III (Continued) 50 l) Planted Culture Examinations II III August 25, 1955 Unclassified spheri- cal blue-green cell Scenedesmus Sp. Aphanocapsa Sp. November 18, 1 955 Chlamydomonas Sp. Chlorococcum Sp. Chlorella Sp. August 25, 1955 Protococcus-like Spores November 19, 1955 Chlorococcum Sp. Encysted Chlamydo monas Sp. August 25 , 1955 No living cells ob- served November 19, 1955 Fungal myceli urn unclassified ___ August 25, 1955 Unclassifi ed dark brown Spore November 19 , 1955 Nannoc hloris bacil- laris *— 51 TABLE III (Continued) Tide I) t : . . Da e Wash Time Time Boot Uncultured Flask Duck Cul- , in in , Time . Wash Washing No. No. ture , HZO AlI‘ , , , in Time Examination Plant , Pen Cage Min. 5 Aug. 4 hr. 20 min. 6, Air Sample 1955 6 Aug. 5 0 25" 30" 8. 1955 7 Re- Aug. 5 0 30" 3 wash 8, hrs. 1955 5" STABLE III (Continued) 52 f L 4 1 Planted Culture Examinations II III August 25 , 1955 Unclassified yellow- gre en flagellate November 19 , 1955 Nannoc hloris bacil- laris April 7, 1956 Nannochloris bacil- filaris Chlo-r‘e-Ila sp. Gloeocapsa sp. Colorless flagel- lates August 23 , 1 955 Only bacteria ob- served November 19 , 1955 Protococc us viridis April 7, 1956 Oedogonium Sp. Protococcus viridis Nannochloris bacil- laris Gloeocystis gigas August 25 , 1 955 Arachnoc hloris-like cell Rhizoclonium fon- tanum Unclassified blue- green cell November 19 , 1955 Rhizocloni um fon- tanum April 14, 1956 Rhizoclonium fog- tanum 53 TABLE III (Continued) Tide . . Date. Wash Time Time Boot Uncultured Flask Duck Cul- , in in , Time . Wash Washing No. No. ture , HZO Air , , , in Time Examination Plant . Pen Cage Min. 9 Aug. 0 0 1 5 " 1 Arachnochloris 15, hr. sp. 1955 Epidermal cells from the duck's feet 10 Aug. 0 0 15" 1 Aphanocapsa sp. 15, hr. Oval yellow- 1955 green cell 17p by 20p 11 Aug. 2 hr. 30 min. 15, Air Sample 1955 54 TABLE III (Continued) Planted Cult ure Examinations I II III August 26, 1955 December 7, 1955 Nothing observed SJJhaerocystis Schroeteri Gloeocystis gigas Tetraedron minimum Bacteria August 26, 1955 December 7, 1955 Nothing observed Protococcus viridis Sphaerocystis Schroteri Tetraedron minimum Oedogonium Sp. Unclassified green unicell Bacteria August 26, 1955 December 7, 1955 April 14, 1956 Unclassified fungal Chroococcus sp. Oscillatoria sp. Spore Gloeocystis gigas Gloeocystis gigas \ 55 TABLE III (Continued) Tide . . Date. Wash Time Time Boot Uncultured Flask Duck Cul- . in in , Time . Wash Washing No. No. ture , HZO Air , , , in Time Examination Plant , Pen Cage Min. 12 Aug. Fresh Lake Water Aphanocapsa sp. 16, 10 ml. of Tide Solution Microcystis Sp. 1955 added to flask after Euglena Sp. inoculation Oscillatoria Sp. 13 Aug. 5 Tide Exposed Oscillatoria Sp. 16, Lake Water Aphanocapsa Sp. 1955 Microcystis sp. 56 TABLE III (Continued) Planted Culture Examinations II III August 26 , 1955 Nannochloris bacil- laris December 8, 1955 Chlamydomonas globosa Sc enedesmus quadricauda Oscillatoria lim- finetica Euglena proxima Navicula Sp. Amoeba Holotricha ciliates September 15, 1955 fienedesmus arma- tus glamydomonas Sp. W sp. Osc111atoria sp. DiatOms August 24, 1955 AraChnOC hloris- like cell December 8, 1955 Scenedesmus Spp. (2) Scenedesmus arma- 21E Gloeocystis gigas Oedogonium Sp. Phacotus lenticularis Ankistrodesmus convolutus Navicula Sp. Phacus orbicularis P. acuminata P. pyrum Lepocinclis acuta Amoeba verrucosa Rotifer Euchlanis April 14, 1956 Encysted Chlamydo- domonas sp. Oscillatoria Spp. (2) Anabaena sp. Navicula sp. Monas-like f lagel- lates Front onia- like cilliate 57 TABLE III (Continued) Tide Date: Wash Time Time Boot Uncultured Flask Duck Cul- , in in , Time . Wash Washing No. No. ture , H20 Air , , , in Time Examination Plant , Pen Cage Min. 14 Aug. Faecal Sample 16, 1 955 15 Aug. 5 30" 15" 3OH Diatom frus- 17 , tules 1955 Dead brownish- green fila- ments 58 TABLE III (Continued) Planted Culture Examinations II III August 26, 1955 Piece of Elodea leaf Unclassified algal Spores Unclassified proto- zoa Bacteria December 8, 1955 Chlamydomonas Sp. Arthrospira Sp. Spirulina Sp. Phormidium Sp. Oscillatoria Spp. (2) Small phytoflagel- lates Unclassified proto- zoan cysts April 14, 1956 Gloeocystis gi gas Spi rogyra Sp. Oscillatoria Spp. (2) Spirulina sp. Paramecium bur- saria August 2 7, 1955 DGbriS only December 8, 1955 Scenedesmus sp. Ankistrodesmus convolutus Chlamydomonas sp. Encysted green spheres Tetrahedron mini- *mum Phormidium Sp. Navicula Sp. 59 TABLE III (Continued) ‘—— Date- Tide Time Time ’ Wash . . Boot Uncultured Flask Duck Cul- , in m , Time . Wash Washing No. No. ture , HZO Air , , , in Time Examination Plant , Pen Cage Min. 16 Aug. 5 30" 15" 5" Fungal spore 17 , 1955 17 Aug. 5 hr. Air Sample 17, 1955 18 Aug. 5 3oH 3oH 30H 18, 1955 TABLE III (Continued) 60 Planted Culture Examinations II III August 27, 1955 Debri s only December 8, 1955 Chlamydomonas sp. Scenedesmus arcua- t_u_s_ Gloeocapsa sp. Ankistrodesmus convolutus Unclassified phyto— flagellates August 2 7, 1955 Debris Only December 8, 1955 Fungal Spore (Alternaria Sp.) M August 2 7, 1955 Debris only December 30, 1955 Scenedesmus Sp. Carteria multifilis Franceia Sp. Chlamydomonas Sp. (palmellaT stage) Ankistrodesmus convolutus 61 TABLE III (Continued) . Date: Tlde Time Time ' Wash . . Boot Uncultured Flask Duck Cul- , in in , Time . Wash Washing No. No. ture . HZO Air , , , in Time Examination Plant , Pen Cage Min. 19 Aug. 3 hr. 45 min. 18, Air Sample 1955 20 1 Aug. 0 1 1 30" Blood cells (bill) 22, day hr. from leg 1955 injury 21 1 Aug. 0 1 1 5" Arachnochloris- (feet) 22, day hr. like cells 1955 62 TABLE III (Continued) Planted Culture Examinations II III October 12, 1955 Chlorococcum sp. Sphae rocystis sp. December 30, 1955 Sphaerocystis Sp. Asterococcus Sp. Chlorococcum Sp. Palmella Sp. Chrysocapsa sp. April 15, 1956 Oscillatoria Sp. Chlamydomonas Sp. Ani sonema- like flagellate ii V October 14, 1955 Aphanothece Sp. December 31, 1955 Nannoc hloris Sp. n; November 11, 1955 Wmus bijuga Womonas Sp. Wria augus- tiSsima NaVicula sp. E25513 sp. December 31, 1955 Scenedesmus abun- dans Auk-{Stardesmus convulutus Oscillatoria angus— tissima Phormidium Sp. April 20, 1 956 Scenedesmus bijuga Scenedesmus abun- dans Mougeotia Sp. Oscillatoria Sp. Navicula sp. \ 63 TABLE III (Continued) Tide . . Date. Wash Time Time Boot Uncultured Flask Duck Cul- . in in , Time . Wash Washing No. No. ture . HZO Air , , , in Time Examination Plant , Pen Cage Min. 22 2 Aug. 5 1 30" 30" Feet: only (feet) 22 , hr. debris 1955 Bill: Arachno- Chloris-like cells 23 2 Aug. 0 1 2 30" Encysted green (feet) 22 , day hr. unicells , 1955 blood cells, and epider- mal cells from the duck's feet 24 2 Aug. 0 l 2 5" (bill) 22, day hr. 1955 TABLE III (Continued) I Planted Culture Examinations II III Novembe r 9 , 1955 Scenedesmus abun- dans Scenedesmus quad- ricauda Chlamydomonas lobosa glamydomonas pseudoPSr'tyi gillamydomvonas Sp. glamydomonas flicic 01a flMstronSmus Sp. '_I‘_§traedron sp. NEVICUla :p, Euglena minuta .—~ December 31, 1955 Scenedesmus gad- ricauda Scenedesmus abun- dans Ankistrodesmus convolutus Chlamydomonas sp. Tetraedron sp. Phormidium Sp. Navicula sp. November 9, 1955 De brig Only December 31, 1955 Phormidium foveo- latum Microcystis Sp. Plectonema nosto— corum Phormidium sp. ._..\ November 5 , 1955 F ungal mycelium December 31, 1955 Fungal mycelium \ fi—v II? V ..a- 65 TABLE III (Continued) Tide . . Date. Wash Time Time Boot Uncultured Flask Duck Cul- , 1n 1n , Time . Wash Washing No. No. ture , HZO Air , . . in Time Examination Plant , Pen Cage Min. 25 4 Aug. 0 4 30" 30" Epidermal cells 22, hrs. from the 1955 duck's feet Unclassified Spores 26 Aug. Air Sample 22, 1955 27 6 Aug. 5 24 30” 25" No algal cells 23, hrs. 1955 66 TABLE III (Continued) Planted Culture Examinations II III November 5 , 1955 Chlamydomonas Sp. Scenedesmus quad- ricauda Ankistrodesmus Sp. Oscillatoria lim- netica Navicula sp. Zoomastigidina December 31, 1955 Scenedesmus abun- dans Scenedesmus quad- ricauda Chlamydom onas Spp. Chlamydomonas globosa Phacotus-like cell Ankistrodesmus Sp. Tetraedron sp. Oscillatoria sp. Navicula sp. Monas-like flagel- lates November 5 , 195 5 Nannochloris bacil- laris January 6, 1956 Nannochloris bacil- laris Chlorella Sp. April 20, 1956 Fern prothallus Chlorella sp. Anisonema-like cell November 5, 1955 Ankistrodesmus convolutus Scenedesmus bijuga January 6, 1956 Anki strodesm us convolutus Scenedesm us bijuga Protococc us Sp. 67 TABLE III (Continued) Tide . . j F1 Date. Wash Time Time Boot Uncultured ask Duck Cul- , in in , T1me . Wash Washing No. No. ture , HZO Air , , , Plant in Pen Cage T1me Examination Min. 28 5 Aug. 5 13 30" 30" Cosmarium Sp. (s)a (died) 23, hrs. Arachnochloris- 1955 like cell Navicula Sp. Euglena sp. Misc. diatoms and Spores 31 1 Aug. 5 24 1 30” Arachnochloris- (s) 24, hrs. hr. like cell 1955 32 7 Aug. 5 19 2 30" Arachnochloris- (3) (died) 24, hrs. hrs. like cell 1955 Unclassified blue-green unicell 33 2nd Aug. 5 19 4 25" Debris only (5) 3 25, hrs. hrs. (boot fell off 1955 during wash- ing) a(s) a lump (3 to 4 grams) of Sphagnum peat was added to the culture flask. 68 TABLE III (Continued) Planted Culture Examinations I II III November 4 and 9, January 6, 1956 1955 Ankistrodesmus Sp. Scenedesmus bijugg Fungal spores Ankistrodesmus Chlamydomonas Sp. convolutus Chrysidella sp. Chromulina sp. Chlamydom onas sp. October 28, 1955 January 6, 1956 April 25, 1956 Etradesmus wis- . Tetradesmus wis- T. wisconsinense gnsinense consinense Chlorella Sp. Phormidium Sp. Zooflagellate \ October 28, 1955 January 6, 1956 Ankistrodesmus Ankistrodesmus convolutus convolutus ""‘"‘""\ OCtOber 28, 1955 January 6, 1956 Bacteria, Fungal Nothing observed Spore unclaSSified blue- green unicell N 69 TABLE III (Continued) Tide . . Date. Wash Time Time Boot Uncultured Flask Duck Cul- , in in . T1me . Wash Washing No. No. ture , HZO Air _ . . In Time Examination Plant , Pen Cage Min. 34 Aug. Air Sample 24, 1955 35 6 Aug. 5 26 24 30" Debris only (s) 26, hrs. hrs. 1955 36 8 Aug. 5 33 16 30" Gray spores (s) 26, hrs. hrs. 1955 37 2nd Aug. 5 24 16 30" Blood cells (S) 2 26, hrs. hrs. from foot 1955 injury 38 Aug. 30 hr. Air Sample 26, 1955 TABLE III (Continued) Planted Culture Examinations II III October 28, 1955 No living cells ob- served January 6, 1956 Nothing observed October 27, 1955 Fungi Spores Scenedesmus Sp. January 6, 1956 Nannoc hloris bacil- laris Monas-like flagel- lat es October 27, 1955 Fungi Bacteria January 6, 1956 F ungi October 21 , 1955 Fungal spore Alter- January 6, 1956 Nannochloris - like naria sp. cell Anabaena (young filament) October 18, 1955 Anabae na Sp. (young) January 6, 1956 Anabaena sp. 71 TABLE III (Continued) Tide . . Date. Wash Time Time Boot Uncultured Flask Duck Cul- , in in , T1me . Wash Washing No. No. ture , HZO Air , , , in Time Examination Plant , Pen Cage Min. 39 Aug. Sample of Lake Water 26, from pen 1955 72 TABLE III (Continued) Planted Culture Examinations II III October 14 and 15, 1955 Scenedesmus arma- ELIE. Scenedesmus abun- 9.85.12 Ankistrodesmus convolutus Oedcgoniuin Sp. Oscillatoria sp. Fragilaria sp. Navicula sp. Paranema Sp. Cladoceran January 6, 1956 Palm ella- like cells Scenedesmus arma- Lug Scenedesmus lim- netica Misc. diatoms Ankistrodesmus convolutus Closterium gracile Bodo sp. April 25, 1956 Scenedesmus bijuga Ankistrodesmus convolutus Chlorella sp. Mougeotia sp. Tetraedron sp. Chlamydomonas Sp. (encysted) Nannochloris bacil— laris Protococcus viridis Gloeocgpsa sp. Oscillatoria lim- netica Oscillatoria Spp. (2) Navicula Sp. Fungal spore £1.33? naria Sp. Fragilaria Sp. Monas-like flagel- lates 73 TABLE IV ORGANISMS IN WATER PEN, 1955 Date Samples Organisms Found August 24 10 m1. Lake Water Gloeocapsa sp. Cultured Gloeocystis Sp. Oscillatoria Sp. Spi rulina Sp. Diat oms August 27 Planted Slides Green algae: Arachnochloris Sp. Gloeocystis Sp. Blue-green algae: Chroococcus Sp. Others: Arcella Sp. Cladoceran Entosflghon sp. Glenodinium sp. Gomphonema Sp. Monas-like flagellates Peranema Sp. we. SP- Rotifers Diatoms Spores Snails Other ciliate protozoa 4‘ l CHAPTER V CONTROLLED EXPERIMENT. 1956 Modifications In 1956 the research plans were modified in order to obtain more precise data in respect to waterfowl dispersal of algae. The general procedure as outlined in Chapter IV was continued with the following modifications. 1. The media was prepared by placing 24 grams of loam soil into a flask and then adding 25 ml. of water from Wintergreen Lake. Pinches (approximately one-Sixteenth of a teaspoon) of CaCO3 and starch were added to another set of flasks before the addition of the soil. The flasks were then autoclaved for one hour at 15 pounds pressure on each of three consecutive days. 2. The sizes of the Erlenmeyer culture flasks were reduced from the 150 or 200 ml. to 50 ml. 3. A 1:625 (solution of Roccal (active ingredient. alkyl C8H17 to C H dimethyl benzyl ammonium chlorides-10%. Sterwin Chem— 18 37 icals. Inc.. New York) was used for the storage of clean slides and coverslips. 74 75 Figure 7. Wintergreen Lake Showing Filamentous Algal Mat and Lily Pads with Boathouse in Background 76 Figure 8. Ducks in Water Pen 77 4. Each flask was flamed before and after the cotton plugs were removed. Pipettes. slides. and coverslips were also flamed prior to use even though they had been cleaned and stored in Roccal solution. 5. After removal from the water pen. the ducks were har- nessed and hung upon a Clothesline. instead of being. placed in the air cages (Figure 9). At the suggestion of Mr. R. D. VanDeusen (1956) an elastic band was placed over the ducks' eyes to reduce their activity while they were hanging on the Clothesline. This blind did not materially affect the activity of the ducks in these experi- ments. and was discontinued. 6. Clean cheesecloth netting was placed around the ducks when they hung in the air for long periods of time (15 hours or over). Netting was also placed over most of the fingerbowls of boiled pondwater which were exposed to the outside air about five feet above ground to reduce the possibility of micro-organism con- tamination by insects (Figure 10). This may be an important factor depending on the specific conditions under which the research is carried out. The results did not seem to indicate that contamination by insects had occurred. 7. Following the suggestion of Dr. W. E. Wade (1956) the boot-wash time was reduced from 30 minutes to 15 minutes. There 78 Figure 9. Ducks in Harnesses and Fingerbowl Hanging on Clothesline Figure 10. Ducks and Fingerbowl Covered with Cheesecloth Netting Hanging on Clothesline 79 80 was no essential difference in the number or type of organisms found when one foot of a bird was boot-washed for 30 minutes and the other for 15 minutes (Figures 11 and 12). 8. The culture flasks. after being inoculated with the boot washings. were stored in an area where they received indirect light throughout the day (Figure 13) as advised by Dr. E. G. Pringsheim (1956). 9. The temperature of the culture flasks was checked by placing a clean centigrade thermometer in a culture flask plugged with cotton. In this way the temperature of the culture medium was recorded as well as the air temperature where the cultures were stored. The water temperature of the culture flask ranged from 14 degrees to. 26 degrees Centigrade. while the air temperature fluctu- ated from 9 degrees to 34.5 degrees Centigrade. 10. The pH of the flasks was. checked with a glass electrode Beckman pH meter. Two flasks with CaCO3 had a pH of 7.3 and 7.4. After swirling. the reading was 8.3. The pH of the flasks with plain soil was 6.2. After swirling. the pH was 6.8. 11. Microscopic examinations were made of some of the un- cultured washings. In Table V are listed the organisms found. 12. The culture flasks were sampled in the following manner. After the flasks were swirled. a sterile pipette was used to collect 81 Figure 11. Duck with Plastic Boots in Funnel. Side View 82 Figure 12. Duck with Plastic Boots in Funnel. Rear View 83 « — hm 2.\ .. Ont. 1.05.“ 5550!... V. i .. .. tries» Culture Flasks Figure 13. 84 medium along the side of the flask from the surface down to the soil. Two drops were taken from the upper. middle. and lower por- tions of the pipette for sampling. These drops were studied micro- scopically by examining three or more transects under low power and three or more transects under high power. An additional ex- amination was made of any visible growth in the flask. The organ- isms present are listed in Tables VI and VII. 13. In addition to the water pen series. another set of ex- periments was conducted whereby the ducks were placed in a pen Situated in the mud and organic debris along the lake shore (Table VIII and Figure 14). 14. Another improvement over the 1955 procedure was the recording of the environmental conditions. Both relative humidity and air temperature were checked while the experimental ducks were in the air. Wind velocity readings were obtained from the Kellogg Forest Station about three miles from the research area (Table XV). Control Culture Flasks Ten culture flasks containing sterile soil-water medium were examined microscopically along with. the experimental culture flasks. They served as controls to determine if any contamination of the ' flasks had occurred either from the soil-water medium or during Figure 14. Mud PPenV 85 86 the examination period. Some of the flasks showed bacterial and/ or fungal contamination during the eight-month period. However. no algal or protozoan contaminations were observed. Fingerbowls of boiled pondwater were exposed to the air for various periods of time and cultured in soil-water medium. The following organisms were observed living in the cultures.(based on 96 examinations of 32 culture flasks). Green algae: Chlamydomonas sp.. Chlorella ellipsoidea. C. vulgaris. Euglena sp.. Gloeocystis gigas. G. vesiculosa. G. sp.. Nannochloris bacillaris. Oedogonium sp.. Pleodornia californica. Protococcus sp.. Rhizoclonium sp.. Scenedesmus abundans. S. bijuga. _S; quadriCaUda: Sphaerocystis sp.. Ulothrix sp.. and Vaucheria sp. Blue-green algae: Chroococcus sp.. Gloeocapsa sp.. Nostoc SP” Qgcillatoria lacustris. O. Minima. 0. sp.. Pelogloea bacillifera. Ebormidium tenue. and P. sp. Protozoa: Colpoda Sp.. unclassified amoeba. and an unclas- Sified f1agellate. Higher plants: Elodea sp.. Fern prothalia. and Etricularia sp. Table IX lists the organisms observed microscopically in the C ontrol Cultures. 87 Another control study was culturing a lO-ml. sample of the boiled pondwater as used for washing the bills. feathers. and feet of the ducks. Examination on April 14. 1957. revealed no living organ- isms. Winter Data In November of 1956 eight feather samples were obtained from four Mallard ducks. After being in the water pen for one hour on November 2, two ducks were held in the air for ten minutes. Lower breast and undertail covert feathers were removed with sterile for- ceps. They were placed in vials half-filled with boiled pondwater. The vials were Capped. Shaken. and left standing for three hours. Cultures were planted with the feather washings on November 2. 1956. Two examinations were made of these cultures. The organisms pres- ent are recorded in Table X. Ten ml. of lakewater were taken from the water pen and planted in soil-water medium on November 2, 1956. A study was made of the uncultured lakewater showing Oedggnium sp.. £11152- clonium sp.. Anabaena sulacylindricia. Microcystis aeruginosa. Navicula sp., Synedra sp., and a copepod nauplius to be present. The first microscopic examinations of the cultures were made on November 16. 1956. and the following organisms were observed: 88 Nannochloris bacillaris. Oocystis E'gas, Scenedesmus quadricauda, Microcystis aerugé'nosa, Microspora Sp.. Oscillatoria Spp. (2), Ila: vicula Sp., and unclassified diatoms. The protozoa present were Uronema sp., Stylonichia-like ciliate, and an unclassified flagellate. On April 6, 1957, the second examinations were made and these organisms were present: Ankistrodesmus falcatus, Chlamy- domonas sp., Oedogonium sp., Pediastrum Boryanum, Scenedesmus bijuga, Sphaerocystis Schroeteri, Tetraedron minimum, Oscillatoria spp. (3), Navicula sp., and Phacus pleuronectes. Other organisms seen were protozoa: Oxytricha sp., and an unclassified ciliate; nematode: Rhabdolaimusrlike; an unclassified rotifer, and bacteria. The remainder of the feather washings were made on Novem- ber 23, 1956. These washings are described and the organisms are recorded on Table XI. Lakewater directly from the water pen was also examined on November 23, 1956. Microcystis aerllginosa. Navicula sp., 93513: gonium Sp., and a copepod nauplius were seen. 89 TABLE V CLASSIFICATION OF ORGANISMS OBSERVED IN UNCULTURED WASHINGS, 1956 Flask No. Organisms Observed 36 and 37a Chlamydomonas sp. (encysted) Synedra sp. Oocystis Sp. Cosmarium Sp. 41 and 42 Oocystis Borgei Oscillatoria sp. 43 and 44 Fragilaria sp. Unclassified cysts or spores 45 and 46 Fungal spore: Alternaria Sp. 48 and 49 Debris only b . 52 and 53 Fungal spore: Alternaria Sp. 55 and 56 Bacteria (faecal contamination) b 57 and 58 Rhizoclonium sp. (faecal contamination) Fungal Spore: Alternaria Sp. 61 and 62 Fungal spore: Alternaria sp. Unclassified spores or cysts (faecal contamination) aCentrifuged for five minutes. bCentrifuged for ten minutes. Note: Cl6’ an uncultured air sample, showed only debris. I 90 TABLE VI DATA FROM CONTROLLED EXPERIMENTS (WATER PEN) WITH CLASSIFICATION OF ORGANISMS, 1956 Soil- Five Time Air Boot Flask Date . 1n , No Pl nt d Water Minute Water T1me Wash ' a e Medium Wash in Min. in Min. Pen 1 July 12 CaCO3a Tide 30 3o 2 July 12 plain Tide 30 30 3 July 12 CaCO3 45 minute Air Sample 4 July 12 plain 45 minute Air Sample 5 July 13 plain Roccal 30 30 1:1250 aPincheS of CaCO3 and starch added. bNo material added to the soil-water medium. 91 TABLE VI (Continued) Planted C ulture Examinations I II III July 18, 1956 September 25, 1956 December 27, 1956 Bacteria Bacteria Protococcus viridis Scenedesmus abun— dans Bacteria July 18, 1956 September 25, 1956 December 27, 1956 Bacteria Bacteria Gleeocystis gigas Protococcus viridis July 19, 1956 September 25, 1956 December 27, 1956 Bacteria Bacteria ‘ Bacteria July 19, 1956 September 25, 1956 December 28, 1956 Bacteria Vaucheria sp. Bacteria July 20, 1956 September 25, 1956 December 28, 1956 Bacteria Bacteria Bacteria Protozoa: unclas- sified ciliates 92 TABLE VI (Continued) . . Time , 8011- Five , Air Boot Flask Date . 1n . No Planted Water Minute Water T1me Wash ' Medium Wash in Min. in Min. Pen 6 July 13 plain Roccal 30 30 1:1250 7 July 13 1 hour Air Sample 93 TABLE VI (Continued) Planted C ulture Examinations II III July 20, 1956 Protozoa: Monas socialis Bodo-like ciliates Bacteria September 25, 1956 Ankistrodesmus spp. (2) Carteria sp. Chlamydomonas sp. Closteriopsis-like cell Scenedesmus quad- ricauda Oscillatoria Sp. Unclassified diatom December 28, 1956 A. Braunii A. convolutus A. falcatus Chlamydomonas globosa Chlorella vulgaris Scenedesmus bijuga S. quadricauda Oscillatoria lim- netica Unclassified diatom July 26, 1956 Bacteria Fungal hyphae September 27, 1956 Bacteria December 31, 1956 Chlorella vulgaris Nannochloris bacil- laris Oscillatoria minima Euglena sp. Fungal hyphae: Alternaria Sp. Bacteria 94 TABLE VI (Continued) 8011- Five Time Air Boot Flask Date . 1n , N P1 nt d Water Mlnute Water T1me Wash 0' a e Medium Wash in Min. in Min. Pen 8 July 13 plain 1 hour Air Sample 9 July 17 CaCO3 Roccal 30 30 1:625 10 July 17 plain Roccal 30 30 1:625 11 July 17 CaCO3 45 minute Air Sample 95 TABLE VI (Continued) Planted C ulture Examinations II III July 26, 1956 Bacteria September 27, 1956 Chlorella vulgiris Bacteria January 2 , 1957 Chlorella vulgaris Nannochloris bacil- laris Pleodorina califor- nica Euglena Sp. Ba cte ria July 26, 1956 Bacteria September 27 , 1956 Bacteria January 2, 1957 Chlorella vulgaris Euglena sp. Nannochloris bacil- laris Bacteria July 26, 1956 Bacteria Fungal hyphae September 27, 1956 Chlorella vulgaris Nannochloris bacil- 1a ris January 3, 1957 Chlorella vulgaris Bacteria July 26, 1956 Fungal hyphae September 27, 1956 Unclassified blue- green cell January 3, 1957 Euglena sp. Fungal hyphae: Alternaria Sp. Bacteria 96 TABLE VI (Continued) 8011- Five T1me Air Boot Flask Date . 1n . N Planted Water Minute Water T1me Wash 0' Medium Wash in Min. in Min. Pen 12 July 17 plain 45 minute Air Sample 13 July 19 CaCO3 30 5 15 14 July 19 plain 3O 5 15 15 July 19 CaCO3 Roccal 15 15 1:625 97 TABLE VI (Continued) Planted Culture Examinations II III July 12, 1956 Fungal hyphae September 29, 1956 Fungal spore: Alternaria Sp. Bacteria January 3, 1957 Bacteria Contaminated July 31, 1956 Mic rocystis aeru- ginosa Bacteria September 29 , 195 6 Chlorella vulgaris Navicula sp. January 8, 1957 Euglena sp. Fungal hyphae Bacte ria July 31, 1956 Nothing observed September 29, 1956 Plectonema—like filament January 10, 1957 sierra sp- Oscillatoria sp. Bacteria 98 TABLE VI (Continued) Soil- Five T1me Air Boot Flask Date . 1n , N P1 t (1 Water Minute Water T1me Wash 0‘ a“ e Medium Wash in Min. in Min. Pen 16 July 19 plain Roccal 15 15 1:625 17 July 19 plain Roccal 15 Bill 1:625 wash:- ing f 18 July 20 Caco3 1'5 / 99 TABLE VI (Continued) Planted Culture Examinations II III July 31 . 1956 Bacteria September 29 , 195 6 Chlorella vulgaris Nannochloris bacil- laris Rhabdoderma irreg- ulare Scenesdesmus_quad- ricauda January 10, 1957 Chlargydomonas Sp. Nannochloris bacil- laris Scenedesmus abun- gees. S. quadricauda sneeze SP- Microcystis aeru- ginosa Oscillatoria sp. Elissa SP- Bacteria July 31 , 1956 Bacte ri a N September 29, 1956 Chlorella vulgaris Oscillatoria lim- netica Bacteria January 12, 1957 Oscillatoria spp. (2) Bacteria July 31 . 1956 Bacteria September 29, 1956 Bacteria January 12, 1957 Chlamydomonas Sp. ageless sp- Bacteria 7 100 TABLE VI (Continued) 3011- Five Time Air Boot Flask Date . 1n , N Pl nt d Water Minute Water T1me Wash 0' a e Medium Wash in Min. in Min. Pen 19 July 20 CaCO3 30 20 July 20 plain 15 101 TABLE VI (Continued) Planted Culture Examinations II III July 31, 1956 Bacteria September 29, 195 6 Spi rulina Sp. Bacteria January 12, 1957 Oscillatoria lim - netica August 1, 1956 October 2 , 1956 January 12, 1957 Protozoa: Monas Ankistrodesmus Sp. Chlamydomonas sp. Holotrich Gloegcystis Sp. Spp. (2) ciliates Nannochloris bacil- Chlorella Sp. Bacteria laris Gloeocystis gigas Scenedesmus biiuga Tetraedron minimum Unclassified phyto- Nannochloris bac il- laris Tetradesmus Sp. flagellate Arthrospira Jenneri Arthrospira Gomon— Microcystis Sp. tiana Oscillatoria lim- Chroococcus dis- netica persus Oscillatoria Sp. Oscillatoria lim- Euglena sp. netica Navicula sp. Euglena minuta Synedra Sp. Protozoa: unclas- Bacteria sified Holotricha 102 TABLE VI (Continued) A —===== Soil- Five Time Air Boot Flask Date . 1n , N Pl t d Water Minute Water T1me Wash 0' an e Medium Wash in Min. in Min. Pen 21 July 21 plain 30 22 July 21 CaCO3 Tide 15 30 15 103 TABLE VI (Continued) Planted Culture Examinations II III August 1, 1956 Euglena sp. Protozoa: unclas- sified ciliates October 2 , 1956 Ankistrodesmus Sp. Gloeocystis Sp. Nannochloris bacil- laris Scenedesmus bijuga S. quadricauda Tetraedron minimum Unclassified phyto- flagellates Oscillatoria lim- netica Euglena spp. (2) Diatoms (sp.) January 13, 1957 Ankistrodesmus Sp. Chlorella ellipsoidea Chlorella vul aris Dactylococcopsis acicularis Scenedesmus di- morphus Scenedesmus Sp. Tetraedron minimum Oscillatoria lim - netica Oscillatoria sp. Euglena minuta Euglena Sp. Diatoms (3 Spp.) Bacteria August 1, 1 956 Bacteria October 2 , 1956 Bacteria January 14, 1957 Bacteria Fungal hyphae Unclassified spore 104 TABLE VI (Continued) Time 8011- Five . Air Boot Flask Date . 1n , N Pl nt (1 Water Minute Water T1me Wash 0' a e Medium Wash in Min. in Min. Pen 23 July 21 plain Tide 15 3O 15 24 July 23 CaCO3 Tide 3O 30 15 25 July 23 plain Tide 30 3O 15 TABLE VI (Continued) 105 Planted Culture Examinations II III August 3 , 1956 Aphanot hece cas— tagn ei Pelcgigea bacillifera Bacteria Protozoa: unclas - sified flagellate October 2 , 1956 Arthrospira Gomo- tiana Euglena sp. (en- cysted) Oscillatoria Spp. (2) Diatom Unclassified phyto- flagellate Bacteria January 15 , 1957 Arthrospira Jen- ‘ neri Euglena Sp. Oscillatoria spp. (2) Diatoms (2) Protozoa: unclas- sified flagellate August 3. 1956 October 2, 1956 January 18, 1957 Aphanothece cas- Micrmfitis-like Bacteria tagnei cell Bacteria August 4, 1 956 October 2, 1956 January 18, 1957 Debris Oscillatoria Spp. (2) Diatoms Bacteria Euglena sp. Navicula Sp. Phormidium tenue Bacteria 106 TABLE VI (Continued) Soil- Five T1me Air Boot Flask Date . 1n , N P1 nt (1 Water Minute Water T1me Wash 0' a e Medium Wash in Min. in Min. Pen 26 July 23 plain Tide 30 30 Bill wash- ing 27 July 23 CaCO3 Tide 1 hr. 30 15 28 July 23 plain Tide 1 hr. 30 15 29 July 23 CaCO3 Tide 2 hrs. 30 15 TABLE VI (Continued) 107 Planted Culture Examinations II III August 9 , 1956 Gonium sociale Fungal hyphae Protozoa: unclas- sified ciliate Unclassified flagel- late October 2 , 1956 Chlorella vulgaris Nannochloris bacil- laris Chroococcus dis- persus Oscillatoria spp. (2) Euglena sp. (en- cysted) Diatoms Protozoa: Monas- 1ike flagellate Bacteria January 18, 1957 Chlamydomonas sp. Chlorella vufilgfaris Oscillatoria lim- netica Phormidium tenue Navicula sp. Synedra sp. August 9 , l 956 Bacteria October 3, 1956 Bacteria February 2 , 1957 Oscillatoria lim- netica Bacteria August 14, 1956 Fungal spore: Alternaria Sp. Bacteria October 2, 1956 Protozoa: unclas- sified flagellate Unclassified ciliate Bacteria February 2 , 195 7 Oscillatoria ac utis- sima O. limnetica Bacteria August 14. 1 956 Aphanothegg SP. October 4 , 1956 Bacteria February 2, 1957 Bacte ria 108 TABLE VI (Continued) . . Time , 3011- Five . Air Boot Flask Date . 1n , No Planted Water Minute Water T1me Wash ' Medium Wash in Min. in Min. Pen 30 July 23 plain Tide 2 hrs. 30 15 31 July 23 CaCO3 Tide 2 hrs. 30 15 Bill Washing TABLE VI (Continued) L —1 109 Planted Cult ure Examinations II III August 14, 1956 Arachnochloris Sp. Navicula sp. Fungal spore: Alternaria Sp. October 4 , 1956 Chlorella vflggis Scenedesmus quad- ricauda Unclassified phyto- flagellate Euglena Sp. Diatoms Bacteria February 2, 1957 Chlorella ellipsoidea Scenedesmus abun— 9.1295 Oscillatoria lim- netica Euglena Sp. Navicula sp. August 14, 1956 Chlorella vu_lgg.ris Gl oeocystis gigas Nannochloris bacil- laris Scenedesmus abun- __ dans S. quad-ricauda Ebenedesmus Sp. 5§cillatoria spp. (2) Efiglena minuta Navicula spp. (2) Protozoa: unclas- sified flagellates Bacteria Fungal hyphae October 4, 1956 Chlorella vggaris Chlorella sp. Gloeocystis gigas Kirchneriella sp. Scenedesmus hijugg S. gpoliensis Unclassified phyto- flagellate Oscillatoria lim- netica _C_)_. Sp. Euglena sp. Diatoms Protozoa: unclas- sified Holotricha February 7 , 1957 Ankistrodesmus Sp. Oscillatoria lim- netica Oscillatoria Sp. Euglena Sp. Navicula sp. Diatom Bacteria ———— 110 TABLE VI (Continued) Soil- Five Time Air Boot Flask Date . 1n , N P1 nted Water Minute Water T1me Wash 0' a Medium Wash in Min. in Min. Pen 32 July 24 CaCO3 Tide 4 hrs. 30 15 33 July 24 plain Tide 8 hrs. 30 15 34 July 24 CaCO3 Tide 8 hrs. 30 15 35 July 24 plain Tide 8 hrs. 30 15 TABLE VI (Continued) 111 Planted Culture Examinations II III August 15 , 1956 Bacte ria October 5 , 1956 Bacteria February 7, 1957 Oscillatoria lim- netica Bacteria August 15 , 1956 Fungal spore: Alternaria Sp. Bacteria October 5 , 1956 C hlorella vulgaris Oscillatoria sp. Bacteria February 16, 1957 Chlamydomonas Sp. Chlorella vulggris Euglena sp. Bacte ria August 15 , 1956 Bacteria Protozoa: unclas- sified unicell October 5 , 1956 Bacteria February 16, 1957 Anabaena sp. Lyngbya limnetica Oscillatoria sub- brevis August 1 5 , 1 956 Bacteria Protozoa: unclas- sified unicell October 5, 1956 Navicula Sp. Bacteria February 16, 1957 Lyrgbya attenuata Oscillatoria Sp. Fungal Spore 112 V -~ .‘3 TABLE VI (Continued) Soil- Five Time Air Boot Flask Date . 1n . No Planted Water Minute Water T1me Wash ' Medium Wash in Min. in Min- Pen 36 July 25 CaCO3 Tide 16 hrs. 30 15 37 July 25 plain Tide 16 hrs. 30 15 TABLE VI (Continued) 113 Planted Culture Examinations II III August 15, 1956 October 5 , 195 7 February 16, 1957 Protozoa: Oikomo- Chlamydpmonas sp. Chlamydomonas nas termo Phacus sp. globosa Bacteria: Spirillum Euglena Sp. Oscillatoria lim- Sp. Navicula Sp. netica Protozoa: Monas Navicula sp. Sp. Protozoa: Monas- Peranema sp. like flagellate Bacteria f August 15, 1956 F11 ngal hyphae Protozoa: Holotrich ciliate October 6, 1956 Ankistrodesmus Sp. Chlorella vulgaris Nannochloris bacil- laris Tetraedron minimum Euglena sp. Phacus sp. . Navicula Sp. Protozoa: Monas Sp. February 17, 1957 Ankistrodesmus convulutus Chlamydomonas Sp. Aphanocapsa Sp. Oscillatoria sp. Phormidium Sp. Euglena sp. Phacus sp. Navicula Sp. Protozoa: Monas sp. Oikom onas sp. Bacteria g Jan. 114 TABLE VI (Continued) . . Time . Flask Date 3011‘ Five . Alr Boot No Planted Water Minute ertl Time Wash ' Medium Wash a er in Min. in Min. Pen 38 July 25 plain Tide 16 hrs. 30 Bill wash- ing 39 July 25 CaCO3 Tide 24 hrs. 30 15 40 July 25 plain Tide 24 hrs. 30 15 ‘ / i 115 TABLE VI (Continued) Planted C ulture Examinations II III August 16, 1956 Oscillatoria lim- netica . Fungal hyphae October 6, 1956 Nannochloris bacil- laris Scenedesmus sp. Unclassified phyto- flagellate Oscillatoria sp. February 17, 1957 Scenedesmus bijuga Tetraedron minimum Oscillatoria sp. Phormidium muci- cola Egglena minuta August 16, 1956 Fungal hyphae Bacteria October 6 , 1956 GloeocLstis gigas Oscillatoria Spp. (2) Bacteria February 21, 1957 Chlamdomonag Sp. Scenedesmus bijuga Lyngbya limnetica Oscillatoria aggusta O. subbrevis August 16, 1956 Chromulina sp. Crypt oglena pigra October 6 , 1956 Euglena sp. (en- cysted) Navicula Sp. Scenedesmus quad- ricauda Unclassified phyto- flagellate Protozoa: Monas Sp. Unclassified ciliate February 21 , 1957 Chlorella vulgaris Nannochloris bacil- laris Protococcus Sp. Rhabdoderma sp. Scenedesmusjilliga S. quadricauda Oscillatoria lim- netica Euglena sp. Navicula Sp. Protozoa: Monas sp. 116 TABLE VI (Continued) Time Soil- Five , Air Boot Flask Date . 1n . No Planted Water Minute Water T1me Wash ' Medium Wash in Min. in MLn. Pen 41 Aug. 2 Caco3 Nonea 1 hr. 15 15 42 Aug. 2 plain 1 hr. 15 15 j 43 Aug. 2 CaCO3 1 hr. 30 15 / aDetergent wash discontinued for remaining washings. 117 TABLE VI (Continued) Planted Culture Examinations II III August 17, 1956 Arachnochloris sp. Unclassified spore October 6 , 1956 Aphanothe ca nidu- lans Bacteria February 22, 1957 Chlamydomonas sp. Euglena sp. Oscillatoria lim- netica Bacteria August 17, 1956 Fungal hyphae Protozoa: Unclas- sified flagellate October 9, 1956 Chlorella sp. Euglena sp. Diatoms (2 Sp.) Unclassified phyto- flagellate February 25, 195 7 Gloeocystis gi gas Nannochloris bacil- laris Oedogpnium Sp. Oscillatoria lim- netica sees sp- Navicula sp. August 18, 1956 Aphanothece nidu- lans Bacte ria October 9 , 1956 Bacteria Protozoa: unclas- sified flagellate February 2 7, 195 7 Microcystis incerta Oocystis Boggei 118 TABLE VI (Continued) Time Soil- Five , Air Boot Flask Date . 1n . No Planted Water Minute W t T1me Wash ‘ - Medium Wash 3 er in Min. in Min. Pen 44 Aug. 2 plain 1 hr. 30 15 45 Aug. 2 CaCO3 1 hr. 1 hr. 15 46 Aug. 2 plain 1 hr. 1 hr. 15 / vv'v 119 TABLE VI (Continued) Planted Culture Examinations II III August 18, 1956 Fungal spore Bacteria October 9 , 1956 Chlorella sp. Chroococcus minu— tus February 2 8, 195 7 Chroococcus Sp. Gomphonema Sp. Oscillatoria lim- netica Oscillatoria Sp. Bacteria , August 18, 1956 Bacte ria October 9 , 1956 Bacteria March 4, 1957 Oscillatoria sub- bre vis Bacteria August 1 8, 1956 Aphanothece nidu- lanS Bacteria October 11, 1956 Euglena Sp. Microcystis aerugi: nosa Osgi-ll—S-toria tenuis Bacteria March 4 , 1957 Oscillatoria Sp. Tetraedron minimum 120 TABLE VI (Continued) Soil- Five T1me Air Boot Flask Date . 1n , No Planted Water Minute Water T1me Wash ° Medium Wash in Min. in Min. Pen 47 Aug. 2 plain 1 hr. 1 hr. 15 Bill Washing 48 Aug. 2 CaCO3 1 hr. 2 hrs. 15 49 Aug. 2 plain 1 hr. 2 hrs. 15 50 Aug. 8 CaCO3 1 hr. 4 hrs. 15 121 TABLE VI (Continued) Planted Culture Examinations II III August 18, 1956 Apahanocapsa elachista Aphanothece nidu- lans Oscillatoria Sp. Rhabdoderma Sp. October 11, 1956 Chlorella sp. Aphanocapsa Sp. XfihrOSpira Sp. Oscillatoria sp. Euglena Sp. Protozoa: unclas- sified flagellate March 4, 1957 Euglena Sp. GloeogLstis sp. Oscillatoria lim— netica August 19, 1956 Gloeothece linearis Bacteria * October 11, 1956 Chlorella vulgaris Euglena sp. Scenedesmus bijuga Scenedesmus sp. March 5, 1957 Euglena Sp. Scenedesmus bijtgg Bacteria August 19, 1956 N . October 12 , 195 6 Bacteria March 6, 1957 Chlamydomonas Sp. Bacteria August 20, 1956 Bacte I‘ia October 12, 195 6 Arachnochloris- like cell Bacteria March 6, 1957 Oscillatoria sp. 0. subbrevis 122 TABLE VI (Continued) I Soil- Five T1me Air Boot Flask Date , 1n , No Planted Water Minute Water T1me Wash ' Medium Wash in Min. in Min. Pen 51 Aug. 8 plain 1 hr. 4 hrs. 15 52 Aug. 8 CaCO3 1 hr. 8 hrs. 15 53 Aug. 3‘ CaCO3 1 hr. 8 hrs. 15 54 Aug. 3 plain 1 hr. 8 hrs. 15 Bill Washing TABLE VI (Continued) 123 Planted Culture Examinations II III August 20, 1956 Bacteria October 12, 195 6 Bacteria March 27, 1957 Oscillatoria sp. Bacteria Unclassified spores August 19, 1956 Bacteria October 16, 1956 Bacteria March 27, 1957 Oscillatoria sub- bre vis Bacteria August 19, 1956 October 16, 1956 March 27, 1957 Bacteria Bacteria Chlamydomonas Sp. Chlorella vulgaris Nannochloris bacil- laris Oscillatoria lim— netica Bacteria \ j. August 20, 1956 Bacte ri a October 16, 1956 Navic ula Sp. Bacteria March 28, 1957 Euglena sp. Navicula sp. Oscillatoria ac utis- sima Bacteria 124 TABLE VI (Continued) Time Soil- Five , Air Boot Flask Date . 1n , No Planted Water Minute Wat T1me Wash ' Medium Wash er in Min. in Min. Pen 55 Aug. 3 CaCO3 1 hr. 12 hrs. 15 56 Aug. 3 plain 1 hr. 12 hrs. 15 573 Aug. 4 CaCO3 1 hr. 16 hrs. 15 58 Aug. 4 plain 1 hr. 16 hrs. 15 59 Aug. 6 CaCO3 1 hr. 24 hrs. 15 60 Aug. 6 plain 1 hr. 24 hrs. 15 aWashings planted in the remaining cultures in this table were obtained from ducks which had been enclosed in cheesecloth netting while exposed to the air. TABLE VI (Continued) 125 Planted Culture Examinations I II III August 20, 1956 October 16, 1956 March 29, 1957 Bacteria Bacteria Bacteria August 20, 1956 October 16, 1956 March 29, 1957 Bacteria Bacteria Bacteria August 20, 1956 October 16, 1956 March 29, 1957 Bacteria Bacteria Bacteria August 20, 1956 October 16, 1956 March 29, 1957 Bacteria Bacteria Bacteria August 20, 1956 October 16, 1956 March 29, 1957 Bacte ria Bacteria Bacte ria August 20, 1956 October 16, 1956 March 29. 1957 Debri S Bacteria Bacteria \ 126 TABLE VI (Continued) 8011- Five Ttme Air Boot Flask Date . 1n . No Planted Water Minute Water T1me Wash ' Medium Wash in Min. in Min. Pen 61 Aug. 15 CaCO3 1 hr. 32 hrs. 15 62 Aug. 15 plain 1 hr. 32 hrs. 15 Note: Numbers 63, 64, and 65 were not used. TABLE VI (Continued) 127 Planted Culture Examinations II III September 10, 1956 Bacteria October 16, 1956 Bacteria March 29, 1957 Bacteria September 10, 1956 Bacteria Unclassified spore October 16, 195 6 Bacteria March 29, 1957 Bacteria 128 TABLE VII DATA FROM CONTROLLED EXPERIMENTS (MISCELLANEOUS) WITH CLASSIFICATION OF ORGANISMS. 1956 . Time _ Flask Date 8011- in Time Boot No Planted Water Water in Wash ' Medium Air in Min. Pen Aa Aug. 6 CaCO3 1 hr. 15 hrs. 15 B Aug. 6 plain 1 hr. 15 hrs. 15 C Aug. 8 CaCO3 1 hr. 24 hrs. 15 (contami- nated; duck fell on ground) while exposed to air. aWashings planted in these cultures except F and G were obtained from ducks which had been enclosed in cheesecloth netting 129 TABLE VII (Continued) Planted Culture Examinations II III August 25, 1956 Bacteria October 27 , 195 6 Bacteria April 3, 1957 Fungal hyphae Bacteria August 25, 1956 Bacteria October 27, 195 6 Ankistrodesmus sp. Nannochloris bacil- laris Oscillatoria tenuis Synechococcus aeru- ginosus April 3, 1957 Anabaena affinis Ankistrodesmus convolutus Chlorella ellip- soidea Chroococcus lim- -__A August 25 , 195 6 Bacteria October 27, 1956 Bacteria April 5, 1957 Bacteria 130 TABLE VII (Continued) Time . _ . t Flask Date 8011 in Time B00 N P1 t d water Water 1" waSh 0' an e Medium Air in Min. Pen D Aug. 8 plain 1 hr. 24 hrs. 15 (contami- nated) E Aug. 8 plain 1 hr. 24 hrs. 15 Bill washing (contaminated) F Aug. 9 CaCO3 Hutchins 5 min. 3 goose 131 TABLE VII (C ontinued) Planted Cult ure Examinations II III August 25, 1956 Bacteria October 30, 1956 Cylindrospermum Sp. Euglena Sp. April 5, 1957 Anabaena affinis Chlamydomonas Sp. (encysted) Euglena Sp. Protococcus Sp. Bacteria September 11, 1956 Navicula Sp. Bacteria Protozoa: unclas- sified flagellates October 30, 195 6 Euglena Sp. Navicula Sp. Oscillatoria Sp. Scenedesmus bijuga April 5, 1957 Navicula Sp. Oscillatoria Sp. Phormidium muci- cola September 11, 1956 Fungal hyphae Protozoa: Sexto- monas-like flagellate; un- classified flagel- late; unclassified Heliozoan October 30 , 1956 Oscillatoria Sp. Phacus sp. Diatom Protozoa: unclas- sified flagellates (Z spp.) April 5, 1957 Chlamydomonas globosa Euglena gracilis Navicula sp. Oscillatoria Spp. (3) 132 TABLE VII (C ontinued) . Time , Flask Date 8011‘ in Time Boot No Planted Water Water in Wash ' Medium Air in Min. Pen G Aug. 9 plain Hutchins 5 min. 3 goose H Aug. 15 CaCO3 1 hr. 24 hrs. 15 I Aug. 15 plain 1 hr. 24 hrs. 15 J Aug. 15 plain Faecal Sample TABLE VH (Continued) 133 L Planted Culture Examinations II III a) 33W September 1 1 , Oscillatoria sp. Navicula sp. 1956 Scenedesmus Sp. Protozoa: Monas sp.; unclassified flagellate Rotifer: Philodina Sp. October 30, 195 6 Oscillatoria Sp. Palmodictyon Sp. Diat om April 5, 1957 Navicula sp. Oscillatoria tennis 0. subbrevis September 11, 1956 Bacteria: lum Sp. Spi ril - November 1, 1956 Bacteria April 5, 1957 Bacte ria September 11, 1956 November 1, 1956 April 5, 1957 Bacteria Anabaena Sp. Anabaena affinis Oscillatoria Sp. Nostoc sp.? Oscillatoria Sp. September 11, 1956 November 1, 1956 April 5, 1957 Debris Bacteria No check made 134 TABLE VII (C ontinued) Soil- Time Time Boot Flask Date in , No Planted Water Water in Wash ' Medium Air in Min. Pen K Aug. 15 CaCO3 Faecal Sample b . . M Sept. 29 plain 3 hour Air Sample Nb Dec. 13 plain 26 hour Air Sample ‘_.'5“” carried out. b . . . Flask exposed to air where culture examinations were 135 TABLE VII (Continued) Planted Culture Examinations I II ' III tember 11, 1956 November I, 1956 April 5, 1957 cillatoria Sp. Oscillatoria Sp. No check made classified spores Bacteria Protozoa: unclas- sified flagellate IV A December 27, 1956 April 14, 1957 3acteria Nothing observed December 27, 1956 April 14, 1957 Fungal hyphae Fungal hyphae Bacteria 136 ‘ TABLE VIII DATA FROM CONTROLLED EXPERIMENTS (MUD PEN) WITH CLASSIFICATION OF ORGANISMS, 1956 Time . _ . t Flask Date 5011 in Time Boo N Pl t d Water M d 1n Wash 0' an e Medium u Air in Min- Pen 66 Aug. 15 CaCO3 1 hr. 1 hr. 15 j 67 Aug. 15 plain 1 hr. 1 hr. 15 _/ 68 Aug. 15 CaCO3 1 hr. 2 hrs. 15 / L,— 1 TABLE VIII (Continued) —_—_— I 137 f L E T Planted Culture Examinations II III E—f Leptember 7, 1956 ’rotozoa: Scy_to- monas-like flag- ellate Bacteria October 19 , 1956 Euglena Sp. Protozoa: Monas sp.; unclassified ciliate; unclassi- fied flagellate Bacteria March 29, 1957 Anabaena sp. Euglena sp. Oscillatoria tere- briformis If :eptember 7, 1956 Euglena sp. Bacteria ’rotoz oa: unclas- sified flagellates (4 spp.) October 19, 1956 Euglena sp. Oscillatoria Sp. Protozoa: Monas sp.; unclassified flagellates (Z spp.) March 30, 1957 Chlamydomonas Sp. Euglena sp. Oscillatoria lim- netica ieptember 7, 1956 ’rotozoa: unclas- sified flagellate Bacteria October 19 , I95 6 Bact eria March 30, 1957 Anabaena Sp. Arthrospira Sp. Oscillatoria Sp. Phormidium Sp. 138 TABLE VIII (Continued) Time Flask Date 8011- in Time Boot No Planted Water Mud in Wash ' Medium Air in Min. Pen 69 Aug. 15 plain 1 hr. 2 hrs. 15 4”... 70 Aug. 15 plain 1 hr. 2 hrs. 15 Bill Washing / 71 Aug. 17 CaCO3 1 hr. 4. hrs. 15 / A . ”AA—— , .. ..-—~f*‘ TABLE VIII (Continued) 139 Planted Culture Examinations II III I; rptember 8, l 956 rotozoa: Monas sp.; Amoeba radiosa acteria October 19, 195 6 Chlorella Sp. Nannochloris-like cell Protococcus sp. Hyalotheca-like cell Oscillatoria Sp. Euglena Sp. Protozoa: unclas- sified flagellate March 30, 1957 Chlamydomonas S p. (encysted) Oscillatoria Sp. Protozoa: Stzlo- nzc hia— like ciliate Bacte ria teptember 8, 1956 ’rotozoa: unclas- sified flagellate Bacteria: Spirillum Sp.; other Species October 26, 1956 Chlamydomonas sp. Chlorella vulgaris Scenedesmus sp. Oscillatoria Sp. Fragilaria Sp. Navicula Sp. Pleurosigma Sp. Bacteria March 30, 1957 Chlamydomonas Sp. (encysted) Oscillatoria sub- brevis Oscillatoria Sp. Euglena Sp. Diplonesis Sp. Navicula sp. Protozoa: unclas- sified ciliate September 8, 195 6 Bacteria October 26, 1956 Bacteria March 30, 1957 Anabaena affinis Mic roclstis aeru- ginosa Oscillatoria spp. (2) "\ ' V 140 TABLE VIII (Continued) , Time . Flask Date 8011‘ in Time BOOt N Plant d Water Mud in Wash 0' e Medium Air in Min. Pen 72 Aug. 1? plain 1 hr. 4 hrs. 15 / 73 Aug. 17 CaCO3 1 hr. 8 hrs. 15 / 74 Aug. 17 plain 1 hr. 8 hrs. 15 TABLE VIII (Continued) 141 Planted Culture Examinations II III I: .tember 8, 1956 atozoa: Proto- monad; Valkam- p_t_i_i_a_-like amoeba; unclassified Holotricha October 27, 1956 Chlamydomonas sp. Ulothrix sp. Unclassified phyto- flagellate Oscillatoria Spp. (3) Euglena Sp. Protozoa: Valkem- Elia-like amoeba; unclassified ciliate March 30, 1957 Chlamxdomonas Sp. Oedogonium Sp. Gloeocapsa Sp. Oscillatoria spp. (Z) Protozoa: unclas- sified Holotricha ptember 8, 1956 )lpoda sp. acteria October 27 , 195 6 Chroococcus minor Protozoa: Pera- nema- like flagellate Bacteria April 1, 1957 Anabaena affinis Bacteria I‘— eptember 8, 195 6 'rotozoa: unclas- sified flagellate Bacteria October 27, 1 956 Nannochloris bacil- laris Ulothrix Sp. Protozoa: unclas- sified amoeba April 1, 1957 Chlamydomonas Sp. Spi rogyra- like zygospore Ulothrix Sp. Anabaena affinis Oscillatoria Sp. Protozoa: Fronto- gig-like ciliate 142 TABLE VIII (Continued) , Time . Flask Date 8011' in Time Boot No Planted water Mud 1“ was“ ' Medium Air in Min. Pen 75 Aug. 1? plain 1 hr. 8 hrs. 15 Bill Washing / 76a Aug. 19 CaCO3 1 hr. 16 hrs. 15 _/ 77 Aug. 19 plain 1 hr. 16 hrs. 15 / to the air. aWashing planted in the remaining cultures obtained from ducks which had been enclosed in cheesecloth netting while expose d TABLE VIII (Continued) A? “1“ 143 Planted Culture Examinations II III '7‘ ieptember 8, l 956 )ebris October 27, 195 6 Chlorella vulgaris Protococcus Sp. Scenedesmus abun- dans Euglena Sp. April 1, 1957 Chlamzdomonas Sp. Scenedesmus quad- ricauda Oscillatoria Sp. Euglena sp. Protozoa: unclas- sified ciliate September 8, 1 95 6 Bacteria Protozoa: unclas- sified flagellate October 27, 195 6 Aphanocapsa sp. Oscillatoria spp. (2) Euglena sp. Protozoa: Monas sp.; Amoeba radiosa-like cell; OikomonaS-like flagellate; un- classified flagel- late (2) April 2., 1957 Anabaena affinis Chlamydomonas (enc yst ed) Euglena Sp. September 8, 1956 Bacteria Protozoa: unclas- sified flagellate October 27 , 1956 Euglena sp. Glenodinium Sp. Protozoa: unclas- sified amoeba (lobopodia) April 3, 1957 as... sp. (en- cysted) Gloeocystis gigas Fungal hyphae Bacteria 144 TABLE VIII (Continued) . Time , Flask Date 8011- in Time BOOt No Plant d water Mud 1“ waSh ' e Medium Air in Min. Pen 78 Aug. 19 CaCO3 1 hr. 24 hrs. 15 I... 79 Aug. 19 plain 1 hr. 24 hrs. 15 / 80 Aug. 19 plain 1 hr. 24 hrs. 15 Bill Washing TABLE VIII (Continued) 145 -. Planted Culture Examinations II III September 8, 195 6 Bacteria: Sp. Spirill um October 2 7, 195 6 Bacteria April 3, 1957 Anabaena Sp. Euglena sp. Oscillatoria Sp. September 8, 1956 Navicula sp. Protozoa: unclas- sified flagellate October 2 7, I956 Euglena Sp. Glenodinium Sp. April 3, 1957 Chlamydomonas Sp. $351313 SP- Ulothrix Sp. Bacteria September 8, l 956 Navicula Sp. (en- cysted) Protozoa: unclas- sified flagellate October 2 7, 195 6 Euglena Spp. (Z) Navicula Sp. Oscillatoria Spp. (2) Protozoa: unclas- sified flagellate April 3, 1957 Anabaena Sp. Oscillatoria Spp. (Z) Euglena Sp. £93919; SD- M sp- 146 TABLE IX DATA FROM CONTROLLED EXPERIMENTS (CONTROL FLASKS) WITH CLASSIFICATION OF ORGANISMS, 1956 Soil- Fl t ask Da e Water Flask Description No. Planted . Medium Cl July 18 CaCO3 Culture flasks control Cl July 18 plain Culture flasks control / C2 July 19 plain Exposed to air 45 minutes / C3 July 20 plain Culture flasks control _/ C4 July 20 CaCO3 Culture flasks control / TABLE IX (Continued) 147 Culture Examinations I II III August 25, 1956 November 9, 1956 No check made Debris Bacteriaa August 25, 1956 November 9, 1956 No check made Fungal hyphae Bacteria Fungal hyphae August 25, 1956 November 9, 1956 April 19, 1957 Debris Bacteria Bacteria August 25, 1956 November 9, 1956 April 19, 1957 Debris Debris Bacteria Fungal hyphae August 24, 1956 November 9, 1956 No check made Debris Debris a . . Bacteria were recorded only when present in large numbers. 148 TABLE IX (Continued) Soil- 1 k t F as Da e Water Flask Description No. Planted , Medlum C5 July 21 CaCO3 15 m1. of Lake Water C6 July 21 CaCO3 30 minute Air Sample C July 21 plain 30 minute Air Sample 149 TABLE IX (Continued) ‘t r— Culture Examinations II III August 24 , 1956 Chlorella Sp. Navicula sp. Spirulina princeps Protozoa: Oikomo- naS Sp.; Codo- age-ca—like cell; , unclassified cili- ates November 9 , 195 6 Ankistrodesmus Sp. Scenedesmus sp. ArthrOSpira Sp. Oscillatoria spp. (2) Euglena sp. (en- cysted) Diatoms (2 sp.) Protozoa: unclas- sified flagellates (2 sp.) April 19, 1957 Oedggonium Sp. Scenedesmus bijuga Aphanocapsa Sp. Arthrospira Sp. Gloeocapsa calcarea Oscillatoria sp. Phormidium Sp. Pelogloea bacil- lifera Euglena sp. (en- cysted) Navicula sp. Protozoa: W sp.; unclassified ciliate Bacteria August 24 , 1956 Bacteria Fungal hyphae November 9 , 1956 Bacteria April 19, 1957 Bacte ria August 24 , 1956 Bacteria Debris November 10 , 1956 Debris April 20, 1957 Bacteria 150 TABLE IX (Continued) Soil- Flabk Date Water Flask Description No. Planted . Medium C8 July 23 CaCO3 4 hour Air Sample C9 July 23 plain 4 hour Air Sample / C10 July 23 plain 10 ml. Lake Water _-_ ‘WJ— . 151 TABLE IX (Continued) Culture Examinations II III August 22 , 195 6 Bacteria November 10, 1956 Bact e ria Fungal hyphae April 20, 1957 Bacteria: Sp. Spi rillum August 22 , 1956 Fungal Spore: Al: ternaria sp. Bacteria No check made No check made August 22 , 1956 Chlorella vulgaris Protozoa: Parame- cium bursaria Fungal hyphae November 10 , l 956 Chromulina Sp. Oscillatoria Sp. Scenedesmus Sp. Protozoa: Ento- Siphon Sp.; un- classified flag- ellate (3) April 20, 1957 Scenedesmus bijuga Nannochloris bacil- laris Oscillatoria acutis— sima Phormidium Sp. Protozoa: unclas- sified flagellates (2 spp.) 152 TABLE IX (Continued) ' ll Soil- Flask Date Water Flask Description No. Planted . Medlum C11 July 24 CaCO3 10 ml. Lake Water C12 July 24 CaCO3 6 hour Air Sample _/ C13 July 24 plain 6 hour Air Sample ’/ ) 153 TABLE IX (Continued) Culture Examinations II III August 22 , 1956 Chlamydomonas sp. Microcystis aerugi- nosa Protozoa: unclas- sified flagellate Bacteria November 10, 1956 Cosmarium Sp. Palmella sp. Scenedesmus bijuga Scenedesmus Sp. Arthrospira Sp. Oscillatoria Spp. (2) Euglena sp. (en- cysted) Navicula sp. April 20, 1957 Scenedesmus bagga- Chlamydomonas (en- cysted) Arthrospira Sp. Oscillatoria Spp. (2) Navicula Sp. Protozoa: unclas- sified flagellate; unclassified ciliate August 22, 1956 Bacteria Fungal hyphae November 16, 1956 Bacteria April 20, 1957 Chlorella vulgaris Nannochloris bacil- laris Oedgggnium Sp. Sphaerocystis Sp. Protozoa: ColEda Sp. August 22, 1956 Bacteria Fungal hyphae November 16, 1956 Bacteria April 20, 1957 Chlamxdomonas (encysted) Nostoc Sp. Oedogonium Sp. Ulothrix Sp. 154 TABLE IX (Continued) Soil- t Flask Da e Water Flask Description No. Planted , Medium C14 July 25 CaCO3 10 ml. Lake Water C15 July 25 CaCO3 5 hour Air Sample C16 July 25 plain 5 hour Air Sample ’7 TABLE IX (Continued) 155 Culture Examinations II III August 22 , 1956 Protozoa: Cycli- dium sp.; Peranema Sp.; unclassified cili- ate; unclassified flagellate Bacteria: Spi rillum sp. (contaminated) August 22 , 1956 Fungal hyphae November 16, 1956 Bacteria April 20, 1957 Chlorella vulgaris Nannochloris bacil- laris Ulothrix Sp. Fungal Spore: Alternaria August 22, 1956 Bacteria Fungal hyphae November 16 , 1956 Bacteria April 20, 1957 Higher plant: Utricularia Sp. a 1111' I‘ll! j 156 TABLE IX (Continued) Soil- Flask Date Water Flask Description No. Planted . Medium C July 26 CaCO Filamentous Mat; 5 minute Roccal l7 3 1:625 wash C18 Aug. 1 plain 5 minute Tide wash, rinse with sterile water (wash bottle); boO‘t wash 15 minutes ____,___... C19 Aug. 1 plain 1:625 Roccal wash, rinse with sterile water (wash bottle); boot wash 15 minutes TABLE IX (Continued) 157 Culture Examinations II III August 22 , 1956 Euglena sp. Protozoa: Chilomo- nas paramecium; Cyclidium Sp. Bacteria November 16, 1956 Chlamy_domonas Sp. Protococcus-like cell Scenedesmus b_iiuga Scenedesmus Spi—— Oscillatoria Sp. Spirulina Sp. Euglena sp. Navicula Sp. Protozoa: Pera- nema sp. April 20, 1957 Gloeocystis Sp. Scenedesmus bijuga Nostoc Sp. Navicula sp. Nematode (round worm) August 22 , 1 956 Protozoa: Pera- nema granulifera Bacteria: Spirillum sp. November 16, 1956 Chlorella Sp. Euglena Sp. Navicula Sp. Oscillatoria Sp. Fungal hyphae April 20, 1957 Chlamydomonas globosa Navicula sp. Nostoc Sp. August 22 , l 956 Debris November 21 , 1956 Nannoc hloris bacil- laris ’ Protococcus Sp. April 20, 1957 Chlorella ellipsoidea Cyanarcus Sp. NannOchloris Sp. 158 TABLE IX (Continued) Soil- t Flask Da e Water Flask Description No. Planted , Medium C20 Aug. 2 CaCO3 5-1/2 hour Air Sample C21 Aug. 2 plain 5-1/2 hour Air Sample C22 Aug. 2 plain 10 ml. Lake Water TABLE IX (Continued) 159 Culture Examinations II III August 21 , 1956 Bacteria November 21 , 1956 Bacteria Fungal hyphae April 20, 1957 Euglena Sp. (encysted) Gloeocystis Sp. Bacte ria August 21, 1956 Bacteria November 21, 1956 Debris April 26, 1957 Pelogloea bacillifora August 21 , 1956 Oscillatoria Sp. Protozoa: Chilo- monas Sp.; Entosiphon Sp.; unclassified flagellate Bacteria November 27 , 1 956 C hlamydom onaS Isphagnicola Chlamydomonas Sp. Chlorella sp. Scenedesmus bilgga Scenedesmus Sp. Oscillatoria tenuis Oscillatoria Sp. Euglena Sp. (2) Diatoms (2 Sp.) Protozoa: Monas- like flagellate Fungal hyphae April 26, 1957 Ankistrodesmus con- volutus Chlamydomonas Sp. C. Sp. (encysted) aloeocystis ampla Nannochloris bacillaris Scenedesmus arcuatus S. armatus S. bijuga S. dimorphus Stigeoclonium Sp. Tetraedron minimum Unclassified phyto- flagellate Chroococcus limneticus Oscillatoria sp. Phormidium Sp. Euglena Sp. (encysted) Fragilaria Sp. Navicula Sp. 160 TABLE IX (Continued) Soil- Flask Date. Water Flask Description No. Planted , Medium C23 Aug. 2 CaCO3 Culture flask control C24 Aug. 2 plain Culture flask control C25 Aug. 3 CaCO3 10 ml. Lake Water C26 Aug. 3 CaCO3 25-1/2 hour Air Sample C27 Aug. 3 plain 25-1/2 hour Air Sample TABLE IX (Continued) 161 Culture Examinations II III August 21 , 1956 Bacteria November 27, 1956 Nothing observed April 27, 1957 Bacteria August 21, 1956 Bacteria November 27, 1956 Nothing observed April 26, 1957 Bacteria August 21 , 1956 Protozoa: Chilo- monas sp.; un- classified flagel- late Bacteria November 27 , I956 Euglena Sp. (en- cysted) Oscillatoria tenuis Diatoms (2 Sp.) Protozoa: unclas- sified flagellate April 26, 1957 Chlorella vulgaris Euglena Sp. (en- cysted) Navicula Sp. Oscillatoria tenuis August 21, 1956 Fungal hyphae Bacteria November 27, 1956 Chroococcus Sp. Bacteria April 27, 1957 Bacteria August 21, 1956 Bacteria November 27 , 1956 Chlorella vulgaris Gloeocapsa sp. April 27, 1957 Chlorella vulgaris Euglena sp. (en- cysted) Bacteria 162 TABLE IX (Continued) Soil- t Flask Da e Water Flask Description No. Planted . Medium C28 Aug. 3 plain Faecal sample cultured C30 Aug. 3 plain Faecal sample cultured C31 Aug. 5 plain 10 ml. Lake Water TABLE IX (Continued) 163 1 hflE Culture Examinations II III August 21 , 1956 Bacteria December 1, 1956 Chlamydomonas Sp. Euglena sp. Oocystis pusilla Fungal Spore April 27, 1957 Nostoc Sp. Oocystis eremos— phaeria August 21, 1956 Bacteria December 1, 1956 Bacteria April 27, 1957 Ba cte ria August 20 , 1956 Chlorella vulgaris Microcystis aeru- inosa Oscillatoria spp. (2) Navicula Sp. Protozoa: Coleps Sp.; geranema BIL; protozoan CYStS; unclassi- fied ciliate; un- classified flagel- late December 8, 195 6 Ankistrodesmus Sp. Protgcoccus Sp. Scenedesmus ar- cuatus Oscillatoria Sp. Euglena sp. Phacus Sp. Diatoms (3 Sp.) Protozoa: Bodo Sp.; Bodo-like flagel- late; Entosiphon Sp.; Heliochona sessilis; Monas Sp. Rotif er: Philodina- like Fungal hyphae April 27, 1957 Navicula Sp. Oscillatoria lim- netica Selenastrum minu- tum Synedra Sp. 164 TABLE IX (Continued) Soil- Flask Date Water Flask Description No. Planted . Medium C29 Aug. 3 CaCO3 Faecal sample cultured C32 Aug. 6 CaCO3 29 hour Air Sample C33 Aug. 6 plain 29 hour Air Sample C34 Aug. 8 CaCO3 10 ml. Lake Water gum-n. u..-- M'fi‘ ' TABLE IX (Continued) 165 Culture Examinations II III August 21 , 1956 Bacteria December 1, 1956 Oscillatoria Sp. April 27, 1957 Phormidium Sp. Bacteria August 20, 1956 Bacteria December 11, 1956 Bacteria April 27, 1957 Bacteria August 20, 1956 Debris December 11 , 1956 Rhizoclonium Sp. Higher plant: Elodea (young plant) April 27, 1957 Higher plant: Utricularia Sp. August 20, l 956 Chlorella Sp. Oscillatoria Sp. Protozoa: Monas Sp.; Chilomonas paramecium Bacteria: Spirillum sp. December 13 , 1956 Chlamydomonas sp. Oscillatoria Spp. (2) Euglena spp. (2) Phacus sp. Diatom (2 sp.) Protozoa: Monas- like cell April 27, 1957 Chladeomonas (encysted) Chroococcus dis- persus Merismopedia sp. Chromulina sp. Navicula sp. Phacus orbicularis 166 TABLE IX (Continued) Soil- Flask Date Water Flask Description No. Planted , Medium C35 Aug. 8 plain 10 ml. Lake Water C35 Aug. 8 CaCO3 10 ml. Lake Water TABLE IX (Continued) 167 Culture Examinations II III August 20, 1956 Chlorella sp. Gloeocystis gigas Pediastrum BorLa- num Fragilaria sp. Navicula sp. Protozoa: Entosiphon Sp; unclassified ciliate; unclas- sified flagellate December 13 , 1956 Ankistrodesmus Sp. A. convolutus Chlorella Sp. Gloeocystis Sp. Nannochloris Sp. Palmodictyon varium Pediastrum tetras Protococcus Sp. Scenedesmus ar- matus 3. wage S. quadricauda Tetraedron Sp. Desmid Aphanocapsa Sp. Euglena Sp. (en- cysted) Phacus anacoelus Navicula Sp. Diatoms (2 Sp.) Protozoa: Monas Sp.; Halteria sp.; unclassified cili- ate; unclassified flagellate April 27, 1957 Ankistrodesmus fal- catus variety acicularis Scenedesmus bijuga Scenedesmus Sp. W Tetraedron mini- mum Oscillatoria sp. Navicula Sp. Protozoa: Monas Sp.; unclassified amoeba August 20, l 956 Bacteria December 13 , 1956 Bacteria April 27, 1957 Bacteria 168 TABLE IX (Continued) Soil- Flask Date Water Flask Description No. Planted , Medium C36 Aug. 14 CaCO3 10 ml. Lake Water C37 Aug. 15 CaCO3 32 hour Air Sample C38 Aug. 15 plain 32 hour Air Sample TABLE IX (Continued) - -7 f -_ Culture Examinations 169 II III September 10, 1956 Lyngbya sp. Oscillatoria Sp. Protozoa: Chilo- monas parame- cium: Cyclidium Sp.; Peranema Sp.; unclassified flagellate December 13 , 1956 Protococcus Sp. Scenedesmus Spp. (Z) Oscillatoria spp. (2) Euglena sp. Navicula Sp. Synedra Sp. Protozoa: Cyclidium Sp.; Peranema sp. April 27, 1957 Chroococcus dis- persus Navic ula sp. Oscillatoria spp. (2) Protozoa: Pera— nema Sp.; un- classified flag- ellate September 10, 1956 Bacteria Debris December 13, 1956 Bacteria April 27, 1957 Bacte ria September 1 0, 1956 Bacteria Debris December 13 , 1956 Chroococcus Sp. Euglena Sp. (en- cysted) Protococcus Sp. Sphaeroclstis Schroeteri April 29, 1957 Gloeocystis gigas Gloeocystis vesicu- losa Rhizoclonium Sp. 170 TABLE IX (Continued) Soil- Flask Date Water Flask Description No. Planted , Medium C39 Aug. 15 CaCO3 5 ml. Mud and Water C40 Aug. 15 plain 5 ml. Mud and Water TABLE IX (Continued) 171 it: Culture Examinations II IH September 1 0, 1956 Phacus acuminata Phacus Sp. Oscillatoria granu— lata Egg-Te}; elongata Euglena vsp. Protozoa: _I_I__o_1_o_- phyra-like ciliate; unclassified flag- ellate December 13, 1956 Oscillatoria Spp. (4) Euglena sp. Phacus sp. Diatom (2 sp.) Protozoa: Monas- like cell; Vorti- cella sp.; unclas- sified ciliates (2) Nematode (round worm) April 29, 1957 Scenedesmus bijuga Anabaena Sp. Oscillatoria am- phibia Navic ula sp. Diat om Nematode (round worm) September 10, 1956 Microcystis aeru- ginosa Oscillatoria Sp. Navicula sp. Synedra sp. Protozoa: Coleps hirtuS; Cyclidium Sp.; Euplotes sp.; Peranema Sp.; dedium vernale; Trachelophyllum Sp. Rotifers (2 sp.) Bacteria December 14 , 1956 Closterium Spp. (2) Rhizoclonium sp. Unclassified phyto- flagellate Anabaena sp. Oscillatoria Sp. 2.31.. sp- Diatoms (2 sp.) Unclassified zygo- Spores Bristleworm (annelid) Copepod: nauplius Nematode (round worm) April 29 , 1957 Chlamydomonas (encysted) Nostoc sp. Oscillatoria Sp. Phormidium Sp. Navicula Sp. Diatom Nematode (round worm) 172 TABLE IX (Continued) Soil~ Flask Date Water Flask Description No. Planted . Medium C41 Aug. 16 CaCO3 4 hour Air Sample C42 Aug. 16 plain 4 hour Air Sample C43 Aug. 16 CaCO3 Culture flask control C44 Aug. 16 plain Culture flask control TABLE IX (Continued) 173 7:: J 1 Culture Examinations I II 111 September 10, 1956 December 14, 1956 April 29, 1957 Bacteria Bacteria Bacteria September 10, 1956 December 14, 1956 April 30, 1957 Debris Oscillatoria lacus- Chlorella vulgaria tris Oscillatoria lacus- Protococcus Sp. tris Bacteria September 10, 1956 December 14, 1956 May 1, 1957 Debris Debris Nothing observed September 10, 1956 December 14, 1956 May 1, 1957 Debris Debris Fungal hyphae Bacte ria 174 TABLE IX (Continued) *1 Soil- Flask Date Water Flask Description No. Planted . Medium 45 . C Aug. 1? plain 5 ml. of Mud cultured C46 Aug. 17 CaCO3 7 hour Air Sample C47 Aug. 17 plain 7 hour Air Sample TABLE IX (Continued) 175 Culture Examinations II III September 10, 1956 Lyngbya Sp. Oscillatoria gardhii December 14 , 1956 Protococcus Sp. Gloeocapsa Sp. Oscillatoria Spp. (3) May 1, 1957 Oscillatoria sp. O. acutissima Navicula sp. O. articulata Euglena sp. (en- Synedra sp. 9. sp. cysted) Diatom Navicula Sp. Navicula Sp. Nematode (round Protozoa: Chilo- Diatom worm) monas parame- Nematode (round cium; Cyclidium worm) Sp.; Euplotes sp.; Monas Sp.; un- classified Heliozoan September 11, 1956 December 14, 1956 May 1, 1957 Fungal Spore Bacteria Fungal hyphae Debris Bacteria Bacteria September 11. 1956 December 15, 1956 May 1, 1957 Debris Debris Oscillatoria lacus- tris Protozoa: unclas- sified amoeba 176 TABLE IX (Continued) Soil- Flask Date Water Flask Description No. Planted , Medium C48 Aug. 19 CaCO3 16 hour Air Sample C49 Aug. 19 plain 16 hour Air Sample C50 Aug. 20 CaCO3 24 hour Air Sample C51 Aug. 20 plain 24 hour Air Sample TABLE IX (Continued) 177 A Culture Examinations I II 111 September 11, 1956 December 15, 1956 May 1, 1957 Bacteria Bacteria Bacteria September 11, 1956 December 15, 1956 May 1, 1957 Fungal spores Fungal hyphae Fern prot hallis Fungal hyphae Bacteria Fern prot hallis Bacteria September 1 l , 1956 December 15 , 1956 May 1, 1957 Bacteria Bacteria Phormidium Sp. Debris Bacteria September 11, 1956 December 15, 1956 May 1, 1957 Protozoa: unclas- sified flagell ate Lyngbya- like filament Bacteria Gloeocystis Sp. Oscillatoria sp. Phormidium tenuis 178 TABLE IX (Continued) Soil- Flask Date Water Flask Description No. Planted . Medium C52 Aug. 20 CaCO3 5 ml. of Mud cultured TABLE IX (Continued) 179 Culture Examinations I II III September 11, 1956 December 15, 1956 May 1, 1957 Anabaena sp. Scenedesmus spp. Ahkistrodesmus Anabaena sp. falcatus Microcystis aeru— ginosa Oscillatoria spp. (2) Euglena acus variety- ri g} da Euglena sp. Lepocinclis acuta Phacus S}: Diatoms (2 spp.) Protozoa: Monas sp.; Chilomonas paramecium; Coleps Sp: un- classified Heli- zoan; unclassified ciliate; unclassi— fied flagellate ArthrOSpira Sp. Lyngbya Sp. Oscillatoria Spp. (2) Euglena sp. (en- cysted) Phacus Sp. Diatoms (2 Spp.) Protozoa: Fron- tonia-like ciliate; unclassified amoeba; unclas- classified flag- ellate Gastrotrich (annelid) Rotifer: Philodina Sp. A. convolutus Cladophora Sp. Oscillatoria Sp. Euglena Sp. Navicula Sp. Diat om Nematode (round worm ) Copepod 180 TABLE IX (Continued) Soil- FlaSk Date Water Flask Description No. Planted . Medium C53 Aug. 20 plain 5 ml. of Mud cultured / C54 Aug. 20 CaCO3 Culture flask control / C55 Aug. 20 plain Culture flask control // TABLE IX (Continued) 181 Culture Examinations II III September 11, 1956 Rhizoclonium sp. Protozoa: Coleps sp.; Chilomonas December 15 , 1956 Chlamydomonas Chlorella Sp. Closterium Sp. May 1, 1957 Closterium Sp. Oscillatoria Sp. Euglena Sp. (en- paramecium; Pediastrum Sp. cysted) Euplotes Sp.; Pediastrum duplex Nematode Halteria sp. Scenedesmus Spp. Rotifer Rotifer (2) Navicula Sp. Synedra Sp. Diatoms (3 spp.) Ostracod Copepod: naupulus Rotifer September 11, 1956 December 17, 1956 May 1, 1957 Bacteria Bacteria Bacteria Debris September 11, 1956 December 17, 1956 May 1, 1957 Debris Bacteria Debris _— #— TABLE X 182 FEATHER WASHINGS PLANTED NOVEMBER 2, 1956, WITH CLASSIFICATION OF ORGANISMS Culture Examinations Flask . Flask No. Description I II ____,._..- 1 12 feathers November 15, 1956 April 6, 1957 from lower breast of a Navicula sp. Chlamydorggnfl SP‘ Mallard duck Bacteria Scenedesmw’ (1/2” X 2") dans Oscillatoria SP- 0. limnetigfi Euglena sp- Navicula sp- Bacteria /- 2 7 undertail November 15, 1956 April 12, 1957 covert feath— ers from a Mallard duck (3/4H x 3-3/4H) Navic ula sp. Protozoa: Front onia-like ciliate Chlamfiioryfflffi sp- . on a ScenedesmW" Oscillatoria sp- 0. limnetica Navicula sp- . tom Unclassified a t o— Protozoa: EELS-2%; ni.....a_‘.'like cilia ’ unclassifie ciliate Bacteria / TABLE X (Continued) 183 Culture Examinations Flask Flask N . ' D ‘ t' o escrlp ion I II 3 15 feathers November 16, 1956 April 12, 1957 from lower breast of a Navicula sp. Chlamydomonas Sp. Mallard duck Protozoa: unclas- Oscillatoria Sp. sified flagellate Navicula sp. Unclassified diatom Fungal hyphae Bacteria 4 10 undertail November 16, 1956 April 12, 1957 covert feath- ers from a Mallard duck Lyngbya s p. Synura sp. Protozoa: unclas- sified flagellate Unclassified Spore Chlamydomonas Sp. Scenedesmus abun- dans Oscillatoria granu- lata Navicula sp. Protozoa: radiosa Amoeba — .. 184 TABLE XI FEATHER AND OTHER WASHINGS PLANTED NOVEMBER 23, 1956, WITH CLASSIFICATION OF ORGANISMS Time in . . Flask Flask Description Water Time In . No. Air l Pen I 1 Feet washing 1 hr. 1 hr. / 2 Bill washing 1 hr. 1 hr- I / 3 Feet washing 1/2 hr. 1 hr: 185 TABLE XI (Continued) Culture Examinations II III December 17, 1956 Bacteria January 21, 1957 Bacteria Fungal hyphae Unclassified cyst or spore April 14, 1957 Chroococcus mini- Site Oscillatoria lacus- tris Oscillatoria Sp. Trichodesmium lacustre Eugle na sp. Bacteria December 17, 1956 Bacteria January 21 , 1957 Euglena Sp. (en- April 14, 1957 Chlamydomonas Sp. cysted) Chlorella vulgaris Bacteria Nannochloris bacil— laris Scenedesmus Sp. Oscillatoria sp. Phormidium Sp. Euglena sp. December 17, 1956 January 23 1957 April 4, 1957 Fungal spore Gloeocystis vesicu- losa Oscillatoria lim- netica Bacteria Chlamydomonas Sp. Chlorella ellip- soidea Stare sp- Oscillatoria Sp. 186 TABLE XI (Continued) Time in Flask Flask Description Water Tim? in No. Air Pen 4 Bill washing 1/2 hr. 1 hr. / 5 1 hour Air Sample / 6 18 Lower breast feathers from a Mallard duck TABLE XI (Continued) 187 Culture Examinations II III December 17, 1956 Debris January 23, 1957 Chlorella ellip- soidea C. vulgaris Scenedesmus abun- danS S. bi'u a April 14, 1957 Ankist rodesm us 1‘ alcat us Chlamydomonas Sp. Scene desmus abun- dans S. bijuga S. quadricauda Euglena sp. Bacte ria December 17, 1956 Bacteria January 26, 1957 Chlorella ellip- soidea C. vulgaris Scenedesmus biJuE Bacteria April 17, 1957 Chlamydomonas Sp. Scenedesmus abun- S. quadricauda Euglena Sp. Bacteria December 1 7, 1956 Bacteria Fungal hyphae January 31, 1957 Chlamydomonas (encysted) Chlorella ellip- soidea C. vulgaris Scenedesmus quad- ricauda Bacteria April 17, 1957 Chlamydomonas Sp. Nannochloris bacil- laris Scenedesmus quad- ricauda Phormidium tenue Series sp- 188 TABLE XI (Continued) Time in . . Flask Flask Description Water Time 1“ No. Air Pen 7 7 Undertail covert feathers from a Mallard duck / 8 21 Lower breast feathers from a Mallard duck / 9 8 Undertail covert feathers from a Mallard duck / 10 Culture flask control / 189 TABLE XI (Continued) Culture Examinations II III December 17, 1956 Protozoa: unclas- sified flagellate Bacteria Unclassified spore January 31, 1957 Ankistrodesmus Braunii Chlorella vulgaris Scenedesmus abun- S. quadricauda Euglena Sp. (en- cysted) April 17, 1957 Characium Sp. Nannochloris bacil- laris Scenedesmus abun- dans Euglena Sp. December 17, 1956 Bact e ria Fungal hyphae February 1, 1957 Chlorella vulgaris Scenedesmus bijuga Merismopedia tenuissima Navicula Sp. Bacteria April 17, 1957 Scenedesmus bijuga Me rismopedia tenuissima Phormidium Sp. Euglena sp. Navicula sp. December 17, 1956 Bacteria Fungal hyphae February 1, 1957 Mic rocystis incerta Bacteria April 17, 1957 Anabaena Sp. Chroococcus dis- persus December 17. 1956 Debris February 1, 195 7 Nothing obse rved April 17, 1957 Debris CHAPTER VI DISCUSSION A thorough review of the literature concerning the role of waterfowl in the dispersal of algae revealed that there is a lack of direct experimental evidence for the transmission of micro- organisms by waterfowl. It is clearly illustrated with one or two exceptions that this means of dispersal has been largely an assump- tion of most ecologists. Ingold (1953, pp. 137, 148) stated: Waterfowl almost certainly play an essential part in the long-distance dispersal of freshwater aquatic fungi. In spite of the fact that these fungi have no air borne spores. the distribution of individual species is just as wide. if not wider, than that of terrestrial species. This raises the Problem of the dispersal of these fungi from one isolated fresh- water system to another. and there can be little doubt, in spite of the absence of direct evidence. that. as with aquatic plants. water birds play an essential part in their long-distance dis- persal. The dispersal of micro-organisms is also attributed to chance or accident. Gulick (1932, p. 423), referring to Pacific oceanic iSIands. stated: Through their lists of species we have been able to verify the ability of certain rather restricted types of organisms to suffer transportation into such distant Spots by rare and rather accidental means. 190 191 Fritsch (1931. p. 253) also mentioned that an element of chance becomes a factor in the populating of ponds with algae. Palmgren (1926. pp. 593. 594. 595) has pointed out: In this multitude of conditions. and of various possibilities for their combination into complexes. probably lies the chief cause of the accidental characteristic of the conditions of occurrence-the stamp of mere chance. In phytogeographical discussion the notion of "chance" consequently means an effective complex of causes. so consti- tuted that scientific research. for the present at least. is unable to propound the problem of the ultimate essential dependence on natural laws. When it appears to be absolutely impossible to anticipate these phenomena of occurrence. then we may characterize the circumstance referred to as chance. When on the other hand we have been able to predict the occurrence. it must be ascribed to law. Therefore. in the author's opinion. as the unknown becomes known. the various "accidental" or "chance" occurrences used to explain dispersal. will be found to be predictable. These occurrences can then be ascribed to some of the various factors governing dis- persal of organisms in a more orderly fashion. This researchhas demonstrated some of these factors and conditions heretofore attrib- uted to chance in the dispersal of micro-organisms. The hunters' data Sheets and the field collections were used only as a preliminary attack toward an explanation of the dispersal of algae by waterfowl. This preliminary investigation Showed that 192 birds washed in sterile water after being Shot in the field. give us an incomplete representation of the forms which might be externally transported. In gathering field data the investigator was faced with the following problems: 1. The exact location of the bird prior to being flushed and shot is difficult to determine. 2. Even if the exact location from which the bird was flushed were known. extensive sampling would be necessary to determine the qualitative population of the micro-organisms in this habitat. 3. The period of time the bird had been in the air prior to being Shot is either unknown or is usually less than a minute in dura- tion. The organisms removed and cultured from a bird flushed from the water which had been in the air less than a minute only demon- strates that these forms are taken from an environment. This does not determine the time that they remain viable. 4. Frequently the exact place where the bird falls after being shot and the micro-organisms which might become adherent to it from that area are unknown. Acquiring birds in the field for sterile washings would best be accomplished by flushing a bird from a body of water and Shooting it down over dry land. For example. a high grassy bank where there 193 would be little likelihood of finding plankton-type organisms. All at- tempt was made by the author to secure birds under these specific conditions (pp. 31-36). In these situations the plankton organisms washed from the birds were actually carried out of the natural en- vironment by the bird and not by the hunter as he scooped up the bird. The organisms washed from birds shot in the field vary con- siderably in species and in number depending upon many factors. The micro-organisms in the environment from which the bird was taken, the period of time the bird was in the air. the sky conditions. air temperature. relative humidity. and wind velocity could all play an important part in determining which organisms are to be dis- Persed in a given situation. For these reasons the birds used in the controlled experiments were used as a check on the field collected sPecimens. Results from the controlled experiments did not lessen?he value of the field research done previously by Darwin. De Guerne. ' \ s . 1 - Klingle. Irenee-Marie. and others. They were intended to supp e , rstand- merit and clarify their findings and give us a more luc1d unde ' ed with mg 0f Some of the previously unconsidered factors concern the dispersal of micro-organism. W *‘A—’ 194 I had originally hoped that I would be able to determine the micro-organisms a duck could pick up in a certain period of time from a particular body of water. Conclusions based on my experi- mental data indicated that this was not possible. For example. just as many varieties of micro-organisms were attached to the ducks that were in the water pen one hour as to ducks in the water pen for 24 or more hours (Table 111, pp. 63. 64). I had expected that more organisms both quantitatively and qualitatively would be gath- ered with each increase in time interval; i.e.. 1/2, 1. 2, 4, and 8 hours. The relationship of numbers of organisms to time was not demonstrated by the results of the research. Frequently statements in the literature indicated that ducks arise from a body of water with filamentous algae and higher aquatic Plants adhering to their feet. feathers. and bills. The author noted in the controlled experiment that although ducks were removed from the water with Lemna minor. Spirggyri. and other filamentous green algae adhering to their feet, these forms were not present after being hung in the air for ten minutes. When placed in the pen Situated in the mud and organic debris. the feet of the ducks collected a large amount of material. After exposure to the air for one-half hour. the feet appeared per- feCtly clean to the naked eye. Even beneath the toe nails no dirt 195 particles were visible. Perhaps the oily secretion from the feet prevents the adherence of materials. Yet many micro-organisms were present; the algal forms being much the same as those found in the water pen series. The protozoa were much more numerous in the mud series (Table VIII. p. 136). This occurred with essen- tially no wind velocity so birds flying at an air speed of 20 or more miles per hour would be apt to carry the material for even Shorter periods of time. The effect of the detergent Tide upon micro-organisms was briefly studied. The cleansing effect of Tide is attested to by the following statement from a letter written by Mr. Owen Carter (1956): . The Tide solution used to wash the waterfowl would act in two ways to eliminate microorganisms from the skin and feathers of the birds (1) by physical removal of the organisms by means of detergent action. and (2) by cidal action. We would expect the washing procedure you described to be very effective in removing all types of surface micro flora and fauna. Spe- cifically regarding antimicrobial activity. our invitro test Shows that a five-minute exposure to Tide under ordinary washing con- ditions will kill 90-95% of the common Gram (+) and Gram (-) bacteria. We do not have similar information relating to the cidal effect of Tide solutions on algae and protozoa. Ten m1. of lakewater with a piece of the filamentous algal mat which was floating abundantly in the lake. was placed in an equal amount of concentrated detergent solution (proportion: 1/4 cup Of Tide in nine CUPS 0f tap-water). This was Shaken for five min- utes and then poured into a sterile flask of soil-water medium for 196 culturing. Later upon microscOpic examination of the culture the following organisms were found to have survived the detergent wash: green algae-~Ankistrodesmus convolutus, Chlamydomonas Sp., {Gloeocystis giga , Lepocinclis acuta, Oedogonium Sp., Scenedesmus armatus, S. Spp. (2); blue-green algae-Anabaena Sp., Spirulina Sp.; other algae--Arachnochloris-like cell, Navicula Sp., Phacus acuminata, P. orbicularis. P. pyrum, protozoa: Amoeba verrucosa, Frontonia-like ciliate, Monas-like flagellate, and rotifer: Euchlanis (Table IV). Ten m1. of the detergent solution were added to a culture flask which had been inoculated with 10 ml. of lakewater. Later examinations showed the following organisms: Chlamydomonas globosa, Nannochloris Sp., Navicula Sp., Oscillatoria limnetica, protozoa: an amoeba and a Holotricha. In both of the culture flasks containing the detergent solution, growth was more rapid and richer than in the other eXperimental flasks. Growth was eSpecially abundant in the former culture when the organisms were eXposed directly to the concentrated Tide solu- tion (Table IV, p. 73). Although some of the feet of the eXperimental ducks were washed for five minutes with the detergent solution. some organisms appeared in the culture flasks. They were Arachnochloris-like cell, Gloeoclfitis gigas, Protococcus viridis, Rhizoclonium fontanum, 197 Scenedesmus abundans. and an unclassified blue-green unicell. The presence of the organisms could possibly have been contributed to the 30-minute exposure to the air before the boot wash. These ex- periments would indicate that Tide is not an algacide. but on the con- trary it seems to accelerate growth in culture flasks. The author noted that the ducks washed in Tide prior to being placed in the water pen tended to pick up a greater variety of forms than did ducks which were trapped from the wild and placed directly in the water pen. One reasonable explanation would be that the de- tergent removes the oily secretion from the ducks' feet facilitating the adherence of micro-organisms. In another instance when a duck had died in the water pen. more algal forms were found to be adherent than occurred on the live birds. Charles Darwin experimented with severed ducks' feet and found that the larval stage of fresh-water mollusks became firmly attached (p. 14). Perhaps the decrease in the amount of oily secretion as well as the lower temperature and lack of move- ment of the feet of a nonliving duck may contribute to an increase in a_dherence of organisms. “Roccal was also found to be an ineffective algacide. Organisms found in the cultures which had survived a five-minute Roccal wash were: green algae--Chlamydomonas Sp.. Chlorella vulgaris. 198 Nannochloris bacillaris. Rhabdoderma irregulare. Scenedesmus abundans. and S. quadricauda; blue-green algae--Lyngbya Sp.. Micro- cystis aeruginosa. Oscillatoria sp.. Plectonema-like filament; other algae-~Eu lena Sp. Tide was more effective than Roccal in the physical removal of micro-organisms. Much work has been done in respect to the dispersal of air- borne fungal Spores. bacteria. and pollen grains. Studies have also been completed concerning the various environmental factors which affect their numbers in the air: Pady (1957). Feinberg (1949). Ingold (1953), Meier and Lindberg (1935). Zobell (1942). and others. The study of algae and protozoa carried by air currents. however. has been largely neglected. When boiled pondwater having been exposed to the air for various periods of time was cultured (p. 86). my re- sults were very similar to those found by Pushkarew (1913). To expose sterile culture media to the air in the laboratory does not demonstrate the exact nature of micro-organism dispersal by air currents. The cultured forms may have been carried into In the the medium from a dried-up culture in the same room 01‘ fro very table where the cultures were placed. 051.1133 . eXP There was little correlation between the time of the . . , r of the boiled pondwater and the humidity. wind veloc1ty. 31 199 temperature, and sky conditions in respect to the air-borne algae obtained from the cultures. The boiled pondwater exposed to the air for the longer period of time generally, but not always, pro- duced the most algal forms. AS Shown in Table IX, a Six-hour exposure yielded more algal forms than did a 24-hour exposure. Wind velocity, humidity, and air temperature may be more important than the time of exposure. A steady breeze would perhaps keep more organisms aloft than would short gusts of wind at a greater air speed (Table XV, p. 228). Insufficient data were gath- ered to reach any conclusions on the effect of these environmental factors on the algal and protozoan content of the air. Faecal material was collected from several birds and cul- tured under sterile conditions (Tables III, VII, and IX). All of the forms recorded were apparently in a healthy vegetative condition and had multiplied, giving rise to a very rich growth in some of the culture flasks. Organisms found in the field collections of faecal material iring the summers of 1955 (p. 57) and 1957 (p, 35) were: green .gae- ‘Chlamydomonas Sp., Gloeocystisggas, SEirogzra sp.; blue‘ . . - h I‘ seen algae--Arthrospira Sp., Phormidium Sp., Spirulina sp.. 0t e lassified gash-Navicula sp.; protozoa--Paramecium bursaria, unC :)tozoa(~1 Cysts and small flagellates. 200 The following forms were taken from three faecal samples during the summer of 1956: green algae--Chlamydomonas Sp., Oocystis Eremosmiaeria, Oocystis pusilla; blue-green algae--Nostoc Sp., Oscillatoria Sp., Phormidium Sp., and unclassified zooflagellates (pp. 132-135; 164-165). The organisms found compared favorably with the work done 3y Messikommer (1943). His method differed from mine in that his microscopic examinations were made directly from the fresh material without use of cultures. Usually, he did not indicate whether his orms were in a viable condition. Messikommer's findings are listed below: green algae-- Ecrospora quadrata, Oedogonium Spp. (2), Staurastrum cingglum, ‘enedesmus ecornis, Spirogyra sp., Tribonema vulgare; other algae-- :mbella cymbiformis, Epithemia zibra, Fragilaria capercina, Gom- fl mema angustatum, Navicula gracilis, N. radiosa, and a living ate. He also found much debris and pieces of both plant and mal material. Various culture media for use in this research were consid- d and soil-water medium was selected. This medium provided near as possible the natural conditions for the growth Of algae . . d for d Protozoa. Certain artific1al media have been recommerlcle ob.e Lre cultures of particular organisms, but have been found t 201 unsatisfactory for growing mixed cultures. According to Dr. E. G. Pringsheim of Pflanzenphysiologisches Institut, Gottingen, Germany (1946, also personal communication in 1956,), "when only one medium is to be used for the culturing of algae, the soil-water medium would be the best selection. Since nutrient requirements for many Species of algae are as yet unknown, many forms which can not be grown in artificial medium will thrive in soil-water medium." 'In 1955 three or four grams of Sphagum peat were added to several of the flasks (Table III, pp. 67, 68) to increase the acidity in an attempt to encourage new forms to develop. There was very little change in the rate of growth or the forms found. In 1956 a pinch (approximately 1/16 of a teaSpoon) of both CaCO3 and starch was added to half of the soil-water medium flasks to produce a more basic medium (Table VI). The pH readings of the various culture flasks are given in Table XVI (Appendix, p. 232). Algal and protozoan growth in quantity and variation of forms was far superior in the plain medium than in the more basic ones. The bacterial and fungal growth occurred at a greater rate in the basic medium while the algal and protozoan growth was extremely poor (Table VI). Those organisms which can form Spores or cysts and those with a matrix were expected to be favored inthe diSpersal by 202 waterfowl. Some cells such as Chlorella and Scenedesmus might be- come embedded in the matrix of other cells (Gloeocystis). In this way they could be carried externally by waterfowl for greater pe- riods of time. An illustrative point may be made of the phytoplankton of the lakes in the Faeroes (Borgesen, 1903). The more common plankton forms found frequently had a matrix such as Cosmarium, Crucigenia, Cyclotella, Gloeocystis, Raphidium, and Sphaerocystis, Staurastrum, and Xanthidium. These islands are located in the Atlantic Ocean about 400 miles from the coast of Norway. The cultured washings which were made Showed, however, that forms of algae both with and without a matrix and those capable of cyst- or spore-formation were taken from the ducks exposed to the air for Short periods of time. The Spore- or cyst-forming algae such as Chlamydomonas and Euglena were more prominent in the longer air exposure. To observe the growth rates of the various algae and protozoa in the culture flasks under these experimental conditions was very interesting. The succession of protozoan forms was more rapid than that of the algal forms. The greatest variety of protozoa oc- curred usually within the first two to four weeks after inoculation. The algal succession was very Slow and some cultures, once the 203 growth peak was reached, were relatively unchanged quantitatively or qualitatively during the remainder of the nine months of culturing. Rao (1953, pp. 173-175) stated the peak of algal growth in numbers was determined by placing 50 gm. of dried soil in various types of media. These cultures were kept in bright light at all times. Medium D318 of Growth pH Readirgg Molischs' solution 130 5.1 Knop‘s medium 191 7.4 Distilled water 70 6.7 The increase in numbers in a mixed culture was much Slower than that of a pure culture. In general, it was not until after a two- month period that the increase of algal numbers reached a peak and the forms that appeared first were not always present later. Some- times different Species would appear from latent forms after four or five months of, culturing. In pure cultures the rate of growth was usually far more rapid with the maximum peak in numbers oc- curring within less time than two months. Spores, cysts, and single cells were found which could not be classified without being cultured. Therefore, microsc0pic ex— aminations of the uncultured washings did not give a complete 204 analysis of the organisms present, and only a few were made in 1956. De Guerne (Appendix B, p. 237) also observed that some forms appearing in his cultures were either not detected or not identifiable in the uncultured state. Bristol (1920, p. 39) mentioned some of these same difficulties in his research of culturing organisms from soil. A great deal of difficulty was experienced in identifying the algae found in the cultures for various reasons. In the first place, the preliminary treatment of the soils was such as to preclude the possibility of the presence of all algae except in a resting condition [in] the initial stages of the cultures. The length of time taken for the germination of these resting forms varied in individual Species, and for some months the cultures contained largely developmental stages which it was impossible to identify with any degree of certainty. Again, the somewhat abnormal conditions of excessive moisture under which the algae were growing tended to produce forms which in some cases were rather different from those of typical species already de- scribed, and it was necessary to decide whether such variations were the result of these conditions or whether they might per- haps characterize new species or varieties. Therefore, the use of cultures seems extremely important to the author for two reasons: (1) to determine the viability of the or- ganisms found, and (2) to aid in classification of Spores and cysts of algae and protozoa which may produce vegetative cells. Other taxonomic difficulties were experienced as follows: (1) only one or a few cells were observed; (2) no reproductive struc- tur‘es were available for study: and (3) some cultures contained 205 species which varied only slightly from species already described, i.e., Size, thickness of matrix, pyrenoid or flagellum lacking or not visible, and cells such as Scenedesmus not being in their usual coenobium form (cells existed singly, in pairs, triples, and in the regular coenobium of four or eight cells). Blue-green cells of ap- proximately lfl in diameter which might be classified either as bac- teria or blue—green algae were also difficult to identify to Species using the present taxonomic keys (see Bibliography). Separate listings of the organisms cultured from the washings of the feet, bills, feathers, gullets, and faecal material of the water- fowl are given in Table XIII (Appendix, p. 214) as well as a listing of the Specific waterfowl studied (Table XII, p. 213). The organisms found in the research area of Wintergreen Lake are listed in Table XIV. Some forms of blue-green algae (Anabaena, Aphanizomenon, W, etc.) which do "bloom" in a fairly Short period of time With Proper environmental conditions, are known to give off toxic Substances in sufficient quantity to cause poisoning of livestock (Ingram and Prescott, 1954, p. 86). The author does not know if any relationship exists between the large numbers of migrant waterfowl flocking to the small ponds, Watering holes, et cetera, in Texas and the toxic poisoning of livestock. 206 The death of waterfowl (Gray, 1943, p. 39) and fish (Ingram and Prescott, 1954, p. 83) also have been attributed to phytoplankton organisms, some of which are capable of being carried by waterfowl from one aquatic environment to another. CHAPTER VII CONCLUSIONS 1. Various parts of 106 waterfowl from seventeen Species (Table XII) were washed with boiled pondwater; Data from the ex- amination of 41 birds were obtained from the hunters' data sheets and field washings. Twenty-three ducks were used in the controlled experiments of 1955, whereas 42 were used in 1956. Organisms collected by washing the gullet, bill, feathers, and feet, as well as organisms from the faecal material of the ducks, were examined directly and after culturing. 2. The research shows that various micro-organisms can be carried both internally and externally by waterfowl. The contents from the gullets sampled produced good algal growth in culture, whereas only a few of the faecal samples contained viable forms. 3. The length of time the birds were in the water pen prior to boot washing did not seem to be correlated with the various forms of organisms which would adhere to the feet of the ducks. Thus, in general, the ducks in the water pen for one hour yielded 207 208 as many organisms as those ducks which had been in the water pen twelve or more hours (Table VI). 4. Generally the ducks exposed to the air for one—half, one, two, and four hours carried a great variety of organisms in a viable condition. Those in the air eight hours transported some organisms on their feet, but a greater variety were found to be carried in their bills. The birds exposed to the air longer than eight hours yielded very few organisms. In one instance three genera were recorded after the duck was in the air for 24 hours (pp. 69-70). 5. The controlled experiments gave a more lucid understand- ing of dispersal of micro-organisms by waterfowl than did the field data. 6. The morphology of algal cells with respect to their ability to be dispersed is not clear. Cells without Spines and matrix were carried, but encysting, spore-producing, and matrix-producing forms were most commonly found on the waterfowl. 7. Planktonic organisms from the water pen were not picked up as readily by the ducks or at least did not survive after exposure to the air as long as the organisms taken from the mud pen. 8. Plain soil-water medium with a pH between 6.5 and 7.0 gave the best results for algal growth under these experimental 209 conditions. A more basic or acidic medium retarded growth in this research. 9. Findings indicate that waterfowl play a major role in the dispersal of algae and protozoa for Short distances but become less important with an increase in distance between bodies of water. Dispersal over a distance of five hundred or more miles, for ex- ample, may take place under certain conditions but would be rather rare, according to information derived from birds sampled in this research. When considering the vast numbers of migratory birds in existence, however, it seems probable that micro-organisms have been dispersed to distant oceanic islands and that certain algal forms would be favored as implied by Warming (1901-1908). 10. Higher forms of life (rotifers, nematodes, copepods, etc.) were not found in the cultured washings taken from ducks exposed to the air for over two hours. However, these organisms were Present in the lakewater taken from the water pen while the ducks Were there and were cultured in the soil-water medium. 11. Chance happenings in dispersal are explainable, in part, in terms of cell morphology, reproductive structures, the nature 0f the dispersing agent, and environmental conditions. 12- Not only the biological and physical nature of the aquatic eflVironment determines the organisms which are to be found in a 210 given environment but also their modes of dispersal. Although often not considered, these are also important in explaining the distribution of aquatic micro-organisms throughout the world. Future investiga- tions will probably reveal "laws of dispersal" which will Show that one form is less favored in diSpersal than another. Apparently natural selection is Operating not only in reSpect to the physical and biological nature of a given aquatic environment, but also as to which organisms will reach these new environments by various modes of dispersal. Specific organisms are not found in all suitable habitats and restriction may be attributed to competition in reaching the suit able envi ronm ent . APPE NDIXES Additional Tables The Author's Translation: "Sur la dissemination dissemination des organismes d‘eau douce par les Palmipedes" by J. M. de Guerne 211 APPENDIX A ADDITIONAL TABLES 212 r» _r .n. 213 TABLE XII WATERFOWL USED FOR WASHINGS Black duck (Anas rubripes) Blue goose (Chen caerulescens) Buffle-head duck (Bucephala albeola) Canada goose (Branta canadensis) Coot (Fulica americana) Eastern Belted Kingfisher (Mggceryle alcyon) Gadwall (Anas strepera) Goldeneye (Glaucinetta clangula americana) Green-winged teal (AnaS carolinensis) Mallard (Anas platyrhynchos) Purple Martin (Progue subis) Redhead duck (Aythya americana) Ring Billed Gull (Larus delawarensis) Ruddy duck (OxLura jamaicensis) Spotted Sandpiper (Actitis macularia) Common Snipe (Capella Elinago) Wood duck (Aix Sponsa) A total of 106 waterfowl were washed in the field and in con- trolled experimentation during 1955-56. Thirty—two were collected by hunters and nine by the author to provide field data. Twenty- three birds in 1955 and forty-two birds in 1956 were used in the controlled experiments. 214 TABLE XIII MICRO-ORGANISMS FOUND ON WATERFOWL USED IN THE CONTROLLED EXPERIMENTSa L A X 1 TL 1— Feet Green Algae: Ankistrodesmus Braunii, A. convolutus, A. falcatus, Arachnochloris-like cells, Arthrospira Gomotiana, A. Jenneri, Chlamydomonas globosa, C. mucicola, C. pseudopertyi, _(_I_. Sp.,b Chlorococcum sp., Chlorella ellipsoidea, C. vulgaris, _C_. sp.,b ClgovsteriOpsiS-like cell, Dactylococcopsis acicularis, Franceia Sp., Glenodinium sp., Gloeocystis gigas,b Mougeotia Sp., Nannochloris bacillaris,b Oedogonium Sp., Oocystis Borgei, Palmodictyon sp., Protococcus Sp., Rhabdoderma . . . b irregulare, Rhizoclonium fontanum, Scenedesmus abundans, S. dimorghus, S. Luadricauda,b _S_. sp., Sphaerocystis Schroe- teri, Tetraedron minimum, T. wisconsinense, 1. Sp., and Ulothrix Sp. Blue- green Algae: Anabaena affinis, Aphanocapsa Sp., Aphanothece castagnei, A. nidulans, Chroococcus dispersus, _C_. minutus, Gloeocapsa Sp., Gloeothece linearis, Lyngbya attenuata, aBacteria have been excluded in the table. bThe most common forms found on the feet. 215 TABLE XIII (Continued) J L. limnetica, .1: sp., Microcystis aeruginosa, Nostoc Sp.(?), Oscillatoria angustissima, O. limnetica, O. subbrevis, O. tenuis, O. terebriformis, Q. Sp.,b Pelogloea bacillifera, Phormidium mucicola, P. tenue, 1:. Sp., Plectonema nosto- c orum , Synechococcus aeruginosus Euglenophyta: Euglena gacilis, E. minuta, E. sp.,b Phacus sp. Chrysophyta: Gomphonema Sp., Navicula Sp.,b Sypedra Sp. Protozoa: Anisonema-like cell, Bodo-like cell, Carteria multifilis, C. Sp., Chromulina Sp., Chrysidella Sp., Cosmarium sp., Cr'ygploglenagpigra, Holotricha ciliate, Monas Sp., Monas-like cell, Oikomonas Sp., Peranema Sp., Phacotus-like cell, Scytomonas-like flagellate, Stjlonchia-like cell, unclassified Heliozoan. Fungi: Alternaria Sp. Rotifer: Philodina Sp. Feathers Green Algae: Ankistrodesmus Sp., Characium sp., Chlamz- domonas sp., Chlorella ellipsoidea, C. vulgaris, g //._ bThe most common forms found on the feet. 216 TABLE XIII (C ontinued) I l, ‘_,___. m Nannochloris bacillaris, Scenedesmus abundans, S. bijuga, S. quadricauda Blue-green Algae: Anabaena sp., Chroococcus dispersus, ijbya Sp., Merismopedia tenuissima, Microcystis incerta, Oscilla- toria granulata, O. limnetica, Q. Sp. Chrysophyta: Navicula sp., unclassified diatom, Synura Sp. Euglenophyta: Euglena Sp. Protozoa: Amoeba radiosa, Frontonia-like ciliate, unclassified ciliate and flagellate Fungi: hyphae Bills Green Algae: Chlamydomonas Sp., Chlorella vulgaris, Nannochloris bacillaris, Oocystis pusilla, O. eremOSphaeria, Protococcus Sp., Scenedesmus abundans, S. bijuga, S. quadricauda, _S_. Sp. Blue-green Algae: Anabaena Sp., Aphanothece Sp., Nostoc Sp. (few cells), Oscillatoria subbrevis, Q. Sp., Phormidium mucicola, P. sp. Chrysophyta: Arachochloris-like cell, Diplonesis sp., Fragilaria Sp., Navicula sp., Pleurosigma Sp. 217 TABLE XIII (C ontinued) J J Euglenophyta: Euglena sp., Phacus Sp. Protozoa: unclassified ciliate and flagellate Gullets Green Algae: Ankistrodesmus convolutues. Gloeocystis Sp., Mougeotia Sp., Nannochloris Sp., Spirogyra Sp. Chrysophyta: Cyclotella Sp., Pleurosigma Sp., Navicula sp. Euglenophyta: Euglena sp., Lepocincles Sp. Protozoa: Chromulina Sp., Monas-like flagellate Rotifer: Bdelloidea. Higher plant: Potomogeton Sp. Faecal Material Green Algae: Chlamydomonas Sp., Gloeocystis gigas Blue-green Algae: Arthrospira Sp., Oscillatoria Sp., Phormidium Sp., Spirulina Sp. Protozoa: Paramecium bursaria, unclassified flagellates 218 TABLE XIV PLANKTONIC MICRO-ORGANISMS TAKEN FROM WINTERGREEN LAKE DURING THE SUMMER OF 1956 -- QUALITATIVE STUDY Water Pen Plankton Bloom Sample, July 20 Blue-green Algae: Anabaena affinis,a A. Bornetiana(?), Chamaesi- phon incrustans, Chrococcus Sp., Lyngbyg Higronymusii, _I__._. a a Sp., Microcystis aeruginosa. Oscillatoria lacustris, 0. lim- netica, 9. sp. Euglenophyta: Euglena Sp. Chrysophyta: Navicula Sp. Fungi: Alternaria Sp. spores Rotifer: unclassified cells Other organisms: ostracod Slides planted in the Water Pen 3-6 inches below the surface of the water on July 2; removed and examined July 15 Green Algae: Chlorococcum Sp., Closterium Sp., Cosmarium sp., Oocystis Sp., Pediastrum Boryanum, Scenedesmus quadri- cauda, Sirggonium sticticum, Spirogyra Spp. (2) a . . Organisms cauSing bloom. 219 TABLE XIV (C ontinued) Blue-green Algae: Anabaena Sp., Microcystis aeruginosa, Oscilla- toria Sp., Rivularia Sp. Euglenophyta: Euglena Sp. Chrysophyta: Fragilaria Sp., Navicula Sp., Synedra sp. Protozoa: Arcella vulgaris, Bodo Sp., Frontonia sp., Loxodes Sp., Pgranema sp., Stylonychia sp., Valkomphia-like amoeba Other organisms: gastrotrich, ostracod Slides planted Juli}: 15; removed and examined for 23 Green Algae: Chlamydomonas sp., OedoLonium Spp. (2), Oocystis Sp., Pediastrum tetras var. tetraodon, 2. sp., Rhizoclonium Sp., Scenedesmus Sp., Sphaerocystis Schroeteri, Staurastrum Sp. Blue-green Algae: Anabaena Sp., Chroococcus Sp., MicrocLstis aeruginosa. Oscillatoria Sp., Synechococcus aeruginosus ChrySOphyta: Arachnochloris Sp., Cyclotella Sp., Cymbella Sp., Fragilaria Sp., Gomphonema Sp., Navicula Sp., Surirella Sp. Protozoa: Amphileptus sp., Arcella Sp., Stentor Sp., Vorticella Sp-. unclassified Heliozoan 220 TABLE XIV (Continued) km: A Slides planted August 3; removed and examined Aggiist 20 Green Algae: Apiogystis Sp., Coleochaeta orbicularis, C. scutata, Cosmarium Sp., Pediastrum Boryanum, P. tetras, 1:. sp., Rhizoclonium Sp., Scenedesmus Sp. Chrysophyta: Gomphonema sp., Navicula Sp., Stauroneis Sp., Synedra Sp. Blue-green Algae: Anabaena Spp. (2), Calothrix sp., Gloeotrichia Sp., Merismopedium Sp., Microcystis aeruginosa, Oscillatoria Spp. (2). Euglenophyta: Euglena sp. Protozoa: Actinoghrys Sp., Arcella sp., Epistylis Sp., LionotuS Sp., Stentor Sp., Valkamphia—like amoeba Other organisms: amphipod, Cladoceran, flatworm (Planaria), gastrotrich, rotifer (Bdelloidea), roundworm (nematod). Plankton net tow preserved in 6-3-1 (Transeau's) solution on Jug 8 (1 gallon of lakewater) Green Algae: Eudorina alegens, Pediastrum integrum, Pleodorina californica. Spirggyra Sp., Volvox_tertius Blue-green Algae: Anabaena affinis, A. Sp., Microcystis aeruginosa Other organisms: Ceratium hirudinella, c0pepod nauplius, rotifer 221 TABLE XIV (Continued) Plankton net tow July 8 (planted in CECOJ culture) Green Algae: Pleodorina californica, Spirpgyra Sp. Euglenophyta: Euglena sp. Bacteria: Spirillum Sp. Other organisms: Chromulina Sp., two unclassified phytoflagellates Plankton net tow (planted in culture media) July 8 Green Algae: Chlamydomonas globosa, 9. sp., Pandorina sp., Scenedesmus Sp., Spirogyra Sp. Blue—green Algae: Anabaena sp., Aphanizomenon flos-aquae, Micro- gystis aeruginosa, Oscillatoria sp. Euglenophyta: Euglena sp., Lepocinclis acuta ChrySOphyta: Navicula Sp. Protozoa: Peranema Sp. Other organisms: unclassified phytoflagellates flankton net tow--July 21 Green Algae: Oedogonium Sp., Pandorina morum, Spircgyra Weberi, gaurastrum pentacerum var. tetracerum, S. Sp., Tetraedron sp. 222 TABLE XIV (Continued) 1 t Blue-green Algae: Anabaena affinis, Microcystis aeruginosaa Chrysophyta: Cyclotella Sp., Navicula Sp. Other organisms: Cladoceran, copepod, rotifer (3 Spp.) Plankton net tow--July 23 Green Algae: Eudorina elegans, Oedqgonium Sp., Oocystis Sp., Pleodorina californica, SJiirogyra Weberi, Staurastrum pen- tacerum var. tetracerum Blue-green Algae: Anabaena affins, Chroococcus dispersus, Micro- cystis aeruginosa Other organisms: Ceratium hirudinella, rotifer (2 Spp.) Plankton net tow--July 24 Green Algae: Pandorina morum, Pleodorina californica, Spirogyra Weberi, Staurastrum pentacerum var. tetracerum, S. sp. Blue-green Algae: Anabaena affinis, A. Sp., Merismopedia Trolleri, Microcystis aeruginosa Other organisms: copepod nauplius, rotifer: Keratella Sp., Bay- cella Sp. a . . Organisms cauSing bloom. 223 TABLE XIV (Continued) Plankton net tow--July 25 Green Algae: Oedogonium Sp., Oocystis Sp., Pandorina morum, Pleodorina californica Blue-green Algae: Anabaena affinis, A. sp., Chroococcus dispersus, Microcystis aeruginosa ChrySOphyta: Navicula Sp. Other organisms: Ceratium hirudinella, rotifer: Keratella Sp., Bgycella Sp. Plankton net tow—-August 2 Green Algae: Oocystis Sp., Pleodorina californiga, Staurastrum Sp. Blue-green Algae: Anabaena affinis, Microcystis aeruginosa,a _Q_S_- cillatoria tenuis Other organisms: Ceratium hirundinella,a copepod nauplius Plankton net tow-~August 3 Green Algae: Spirggyra Weberi Blue-green Algae: Microcystis aeruginosa Other organisms: Ceratium hirundinella, Cladoceran Plankton net tow--August 6 Green Algae: Oedogonium sp., Spirogyra Weberi ‘ a . . Organisms causmg bloom. 224 TABLE XIV (Continued) Blue-green Algae: Gloeotrichia Sp., Microcystis abundans, Nostoc punct if orme Chrysophyta: Navicula Sp. Plankton net tow-—August 9 Green Algae: Oocystisfipyriformis, Pandorina morum, Pediastrum duplex, Pleodorina californica, Volvox tertius Blue-green Algae: Anabaena affinis, A. Sp., Chroococcus dispersus, Microcystis aeruginosa ChrySOphyta: Navic ula Sp. Protozoa: Arcella vulgaris Other organisms: Ceratium hirundinella, rotifer Plankton net tow-~Auggst 8 Green Algae: Pandorina morum, Pleodorina californica,a Rhizoclo- nium Sp., Sphaerogystis Schroeteri Blue-green Algae: Anabaena affinis,a A. subcylindraca, A. Sp., . . . a Microcystis aeruginosa Chrysophyta: Navicula SP- Protozoa: Diffulgia sp. Other organisms: copepod nauplius, rotifer (3 Spp.) __m a . . Organisms causmg bloom. 225 TABLE XIV (Continued) Plankton net tow-"August 14 Green Algae: Pandorina morum, Pleodorina californica,a Sphaero- cystis Schroeteri Blue-green Algae: Anabaena affinis,a A. Subcylindrica, A. sp., . . . . ' a . . Chroococcus diSpersuS, Microcystis aeru inosa, OSCillatorla Sp. Chrysophyta: Navicula Sp., Syfledra Sp. Fungi: Alteinaria Sp. Spore Other organisms: Ceratium hirudinella, rotifer: Keratella BrLcella Sp. , Brachionus sp. Filamentous Mat of Algae Floatirg near the Water Pew-Living and Preserved Material, July 9 Green Algae: Oedogonium Sp., Oocystis Sp., Pediastrum Boryanium, Rhizoclonium sp., Spirogyra Spp. (2) Blue-green Algae: Anabaena affinis, A. sp., Aphanizomenon flos- aguae, Aphanocapsa Sp., Aphanothece gelatinosa, A. micro— Spora, Microcystis aeruginosa, Oscillatoria Spp. (2) EuglenOphyta: Etiglena Sp. ChrySOphyta: Gomphonema Sp., Navicula Sp. a . . Orgamsms causmg bloom . 226 TABLE XIV (Continued) Rotifers: unclassified cells Protozoa: not recorded Mud Pen Five- milliliter sample- -August 16 Blue-green Algae: Anabaena affinis, Chroococcus minimum, lim- bya Sp., Microcystis aerpginosa, Oscillatoria Sp. Euglenophyta: Phacus orbicularis ChrySOphyta: Cocconeis Sp., Navicula Sp., Synedra Sp. Fungi: Alternaria sp. Spore Five- milliliter sample- - Afiuggt 17 Green Algae: Cerasterias sp. Blue-green Algae: Anabaena affinis, Aphanocapsa pulchra, Chroo- coccus Sp., Microcystis aeruginosa, Oscillatoria Sp. Euglenophyta: Phacus sp. Chrysophyta: Cocconeis Sp., Navicula Sp. Fungi: Alternaria Sp. spores Other organisms: nematode Five-milliliter sampleuAtgujt 19 Green Algae: Ankistrodesmus Sp., Pandorina morum, Scenedesmus quadricauda 227 TABLE XIV (Continued) Blue-green Algae: Anabaena affinis, Lingbya limnetica, Microcystis aeruginosa, Oscillatoria lacustris, (_)_. Sp.,_Synechococcus aeruginosus Chrysophyta: Cocconeis Sp., Navicula Sp., Stauroneis Sp. Protozoa: Holotrich ciliate Fungi: Alternaria sp. spore v—v fi TABLE XV ENVIRONMENTAL FACTORS IN THE CONTROLLED EXPERIMENTS, 1956a 228 Organisms Time R1318- Sky Air Wind VelocityC Total of tlve . D t (Genera) Ex- Hu- Con- Temp. Miles ae from Air po_ midfiy di-b Max.- 8:00 1:00 5:00 of Sample sure (pct.) tion Min. a.m. p.m. p.m. Wind 7/12 Vaucheria 45 75-88 C 62-80 1 4 4 36 min. 7/13 Chlorella 1 . PC .62-79 2 4 z 36 Nanno- hr. chloris Pleodorina Oscillatoria Euglena 7/17 Euglena 45 66 PC 59-77 2 4 1 38 min. 7/19 nothing 45 77 C 57-75 1 0 4 24 min. 7/21 nothing 30 72 PC 63-83 2 3 2 20 min. aBacteria and fungi are not listed. b c: cWind velocity data were obtained from W. K. Kellogg Fore“ Station. cloudy; PC = partly cloudy; C1 = clear. 229 TABLE XV (Continued) f W. I #47:: Organisms Tyre fig: Sky Air “Find Velocity Total Date (Genera) Ex- Hu- Con- Temp. Miles from Air po- midity di- Max.- 8:00 1:00 5:00 Of Sample sure (pct.) tion Min. a.m. p.m. p.m. “find 7/23 nothing 4 59-63 PC 59-79 1 3 2 31 hr. 7/24 Chlamydo- 6 51-63 CI 59-89 2 2 2 24 (d) monas hr. Chlorella Nanno- chloris Oedogonium Sphaero- cystis Ulothrix Nostoc Colpoda 7/25 Chlorella 5 72-82 C 66-81 1 l 0 17 Nanno- hr. chloris Ulothrix Utricularia 8/2 Euglena 5- 48-56 CI 54-77 3 z 5 38 Gloeocystis 1/2 Pelogloea hr. . dCheesecloth netting was placed over the remainder of the h ngerbowls . TABLE XV (Continued) 230 i , Time Rela- . . Organisms of tive Sky Air “find Veloc1ty Total Date (Genera) Ex- Hu— Con- Temp. —fi Miles from Air po— midity di- Max.- 8:00 1:00 5:00 of Sample sure (pct.) tion Min. a.m. p.m. p.m. “Find 8/3 Chlorella 25- 51-87 C 55-77 1 1 l 31 Chroococ- 1/2 cus hr. Gloeocapsa Euglena 8/6 Rhizoclo— 29 86-90 C 64-81 2 3 l 31 nium hr. Utricularia 8/15 Gloeocystis 32 44-90 PC 61—85 1 1 2 19 Protococ- hr. cus Rhizoclo- nium Sphaero- cystis Chroococ— cus Euglena 8/16 Chlorella 4 54-61 PC 65-87 1 4 0 22 Oscillatoria hr. Protococ- cus TABLE XV (Continued) 231 , Time Rela- . . . Orgamsms of five Sky Air Wind Veloc1ty Total D t (Genera) Ex- Hu- Con- Temp. Miles ae from Air pm midfiy di- Max.- 8:00 1:00 5.00 of Sample sure (pct.) ion Min am pm pm Wind 8/17 Oscillatoria 7 59-70 PC 63-88 2 2 1 15 UnclaS- hr. sified amoeba 8/19 Fern pro- 16 53-78 PC 55-76 - 4 1 45 thallus 8/20 Gloeocystis 24 53-78 PC 48-70 2 0 1 33 Lyngbya- hr. like cell Oscillatoria Phormid- ium Unclas- sified flagellate g: h TABLE XVI pH OF CULTURE FLASKS, 1956 232 pH pH PH pH Flask before after Flask before after No.a Swirl- Swirl- No.a Swirl- Swirl- ing ing ing ing July 30, 1956 CaCO3b 7.3 8.3 Plain 6.2 6.8 CaCO3 7.4 - January 5 , 1957 C 7(c) 7.7 8.1 19(p) 7.4 7.4 8(p) 6.9 6.9 20(c) 6.3 6.5 9(c) 7.9 8.0 21(p) 7.2 7.4 10(p) 7.0 7.1 22(c) 7.3 7.4 11(c) 7.8 8.0 23(p) 7.4 7.4 mm) 7.4 7.4 24(c) 8.3 8.4 17(p) 6.5 6.6 25(p) 7.6 7.7 18(c) 8.0 7.9 26(p) 7.4 7.5 aL(c) = CaCO3; (p) = plain. bColman pH electrometer (Model 18) was used for these readings, cBeckman pH meter (Serial No. 126747) was used for the remainder. 233 TABLE XVI (Continued) pH pH fl pH pH Flask before after Flask before after No. Swirl- Swirl- No. Swirl- Swirl- ing ing ing ing January 5, 1957 27(c) - 8.1 39(c) - 7.5 28(p) 7.1 7.0 40(p) 7.2 7.0 29(c) 8.3 8.3 41(c) 8.0 7.9 30(p) 7.6 7.5 42(p) 7.4 7.4 31 7.4 7.4 43(c) 8.1 8.3 32(c) 8.4 8.4 44(p) 7.3 7.2 33(p) 7.3 7.3 45(c) 8.3 8.4 34(c) 7.8 8.0 46(p) 7.5 7.3 35(p) 7.4 7.4 47(c) 8.0 8.0 36(c) 7.5 7.5 48(c) - 8.3 37(p) 7.2 7.1 49(p) - 7.2 38(p) 7.2 7.2 50(c) 7.3 7.4 March 29, 1957 51(p) 8.1 8.2 53(p) 7.6 7.5 52(c) 8.1 8.1 54 7.3 7.6 234 TABLE XVI (Continued) pH pH pH 35H Flask before after Flask before after No. Swirl- Swirl- No. Swirl- Swirl- ing ing ing ing March 29, 1957 55(c) 7.8 7.8 59(c) 8.3 - 56(p) 7.3 7.1 60(1)) 6.9 6.8 57(c) 8.2 8.2 61(c) 8.2 - 58(p) 7.2 7.0 62(p) 7.6 7.6 April 6, 1957 66(c) 8.1 - 80(p) 7.4 7.5 70(p) 7.5 7.5 C(c) 8.3 8.3 72(p) 7.1 - D(p) 7.4 7.3 74 6.8 6.7 mm 7.4 7.4 75 7.2 7.2 th) 7.7 7.7 79(p) 7.4 7.3 I(p) 7,2 _ Mag 5, 1957 C47th 7.5 6.8 C55(p) 7.5 7.4 C49(p) 7.0 - M m . _ fly: Note: The pH of the water pen during 1956 experimental period varied from 10.0 in June to 7.5 the latter part of August. 235 TABLE XVII FIELD DATA SHEET FOR DUCK HUNTERS Research is being carried out to determine what microscOpic organisms are carried externally on waterfowl from one body of water to another. PLEASE CUT OFF THE HEAD AND FEET OF THE DUCK SHOT AND PLACE THEM WITH THIS DATA SHEET IN THE PAPER BAG PROVIDED. LEAVE IT AT THE ROSE LAKE EXPERIMENTAL STATION OR RETURN TO H. E. SCHLICHTING, ROOM 131, NATURAL SCIENCE BUILDING, MICHIGAN STATE COLLEGE. Your assistance in the collection of material will be greatly appreciated. Please check the appropriate items: Waterfowl shot (check only one): Coot Blue- winged teal Wood duck Mallard Ruddy duck Ring-neck duck Canada Goose Canvasback White-winged Scooter Blue Goose Greater Scaup duck American Scooter Black Duck Lesser Scaup duck Hooded Merganser Baldpate American Goldeneye American Merganser Pintail Bufflehead Red-breasted Merganser Green-winged teal Old Squaw Other (or if in doubt) Sex Of bird: male female The bird was Shot (please check one): coming into the body of water. leaving the body of water. in the water. Name and location of the body of water Time of day that the bird was shot Datefi Name and address of hunter RemarkS: # ‘F APPENDIX B SUR LA DISSEMINATION DES ORGANISMES D'EAU DOUCE PAR LES PALNIIPEDES 236 APPENDIX B ON THE DISSEMINATION OF ORGANISMS FROM FRESHWATER BY THE WEB-FOOTED by Mr. Jules de Guerne1 The possibility of the transport of organisms by the birds is admitted by the majority of naturalists. However, if one begins to look on what basis this opinion almost always rests, we recognize that but for the plants the number of observed facts is extremely limited. Lyell and Darwin, who have specially studied the mode of dispersion applied to plants are far from having unrecognized its importance concerning the aquatic animals. But they did not use the micrOSCOpe, and it is probably the main reason which has prevented them from going deeply into the question. Even in the last years of his life, Darwin was to be preoccupied by it. The more startling observations that we possess on the tranSport of the Lamelibranchs by winged beings, birds and insects, were published by him in 1878 and 1882. 1J. M. de Guerne. Sur la dissemination des organismes d'eau douce par les Palmipedes. Societe de VBiologie. 8 (March, 1888). PP. 294-298. 237 238 Aside from these documents, rather few in number, collected by Darwin, I only know but one precise fact on this subject which was reported by Professor F. A. Forel in 1876 according to Alois Humbert. This naturalist has found sticking to the feathers of ducks and of Grebes some winter eggs of Cladocerans. This question was therefore hardly scratched when upon the return from the third voyage of the Hirondelle, completed under the direction of S. A. the Prince Albert of Monaco, after having dis- covered at the Azores a lacustral fauna composed almost entirely of European types, Spread over a considerable geographical area, I at- tempted to bring about new arguments in favor of this doctrine of tranSport. My researches started in the fall of 1887, at the time of the arrival of birds from the North, and have been continuing during each winter under circumstances more or less favorable. I limited myself, until now, in examining the web-footed, and especially of the COmmon wild ducks, Anas boschas, ordinarily very abundant and easy t0 obtain. Two Species of Teal (Querquedula circia and Q. crecca) and also various birds not Specified, likewise, have furnished me S(”he Objects for study. I have had at my disposal the game coming from the hunting of S- A. the Prince Albert of Monaco, at Marchais, Aisne, and sent ‘6 239 forth directly to Paris. Besides, a well known zoologist, M. Chev- reaux, has beenOkind enough to send me the product of the washing from the feet of several Teal (Querauedula crecca L.) killed at Croisic, (Loire-Inferieure). I have examined as well a certain number of web-footed killed in January, 1888, in the marshes of Arleuz, near Douai (Nord). Finally at different times I myself have procured wild ducks in different parts of Paris, in the markets, or from peddlers. The person in charge Of buying being ignorant of the purpose pursued could not be tempted, consequently, to choose among those birds whose feet appear particularly dirty.. By a trick of fate which iS Certainly permissible to call upon as a favorable argument in this case, that it is on the feet of the duck handled many times from the marsh, to the central market, and at the retail store where I have found some of the most interesting objects. mentioned below (Sytheridea torosa, for example). Except for the Teal of the Croisic Which had been examined immediately by M. Chevreaux, the inspec- tion of the birds had not occurred until nearly twenty-four hours after death. It is on the average the time necessary for the arrival 0f the ducks from the region of the Nord (Bay of Somme, etc.) from place of the hunting to the hands of the consumers in Paris. The State of freshness of the viscera (the digestive apparatus several g. 240 times supplied valuable indications about the last stop of the Bird) Showed me that in several circumstances this delay had not even been attained. Two procedures have been followed for the researches: 1. The direct examination, practiced either immediately on the material collected and diluted with water or at the end of a certain period on the product of the washing of the feet and of the bill in the water with the addition of alcohol immediately after the Operation. 2. The culture of the material collected. The direct observation furnished me the following results. All the webbed-feet examined, with a few exceptions, carried foreign material on various parts of the body. From the point of view of the quantity transported, the feet Should be cited as the most im- POr'tant, next are the edge of. the tongue and the bill, finally the feathers. The latter, oily and compact, appear to be generally very Clean. However, it has happened to me to find a few Spatters on the neck, on the face, and inside of the secondary wing feathers. Those On the wing feathers are produced in all probability when the bird has Shaken itself on the bank or else even in open water. as is SO often the case. The Specks of dirt whose composition I have studied Were entirely made up of microscOpic plant debris. I do not doubt 241 that in the future researches, one will meet such Spatters or of organisms in the state of latent life (Spores, winter eggs, etc.) capable of being tranSported from one lake or from one marsh to another. These Specks have a good hold on the feather while dry, but are dissolved quickly in the water: this circumstance appears to be most favorable to the dissemination. I insist on the transport by the feathers, it is on them, in fact, that can be removed the bodies that float far from the banks on clear and deep water; the question offers a great interest from the point of view of the dispersion of some lacustral pelagic types: but it will be necessary, in order to definitely solve it, to undertake series of observations on places of hunting near large surfaces of water. If the preceding observations allow some doubt as to‘the Subject of transport of pelagic organisms by the birds, those that follow show the important role which these last play in the diS- semination of the littoral forms. AS I have said, it is upon the feet that one finds most of the material carried. On November 18, 1887, it happened to me to gather on the upper part of the membranes between the toes of a Wild duck a quantity sufficient to entirely cover the bottom of a plate 15 centimeters in diameter. It was the murk from one peaty marsh, a bit brown, formed almost exclusively of plant debris (many 242 attaining l centimeter in. length) mixed with a very small number of round quartz grains. In this deposit, I have found the presence of a large number of micrOSCOpic cysts, animal and plant, of many diatoms, of one desmid, of one Cladoceran egg (Lyndeide?), of the half of a Pluetella repens statoblast, and one Ostracod valve. The latter, thanks to a particular definite character, has been able to be determined. It belongs to a Species unknown in France, Cytheridea torosa Jones, but whose geographic distribution is very extensive. They have been reported in England, in the A20 Sea, in the east, etc. It lives in fresh and brackish waters and occurs especially in estuaries. Among these easily recognizable bodies are found many others which the specialists may succeed in naming. The fragments Of insects are numerous. The majority hairy or even thorny in ap- Pearance should easily hook themselves and retain in addition some diverse matter. One of the fragments, a Dipteran femur three mili- meters long, formed a true protective tube where some delicate beings incapable of surviving the dessication could find protection. The compression had forced out some pieces of trachea and a very large number of infusoria cysts. It is worth while to remember this fact: some Similar debris is to be found frequently; in conserv- ing small quantities of water by protection from evaporation, they 243 render perhaps possible the dissemination of certain aquatic organ- isms in their normal surroundings. The case which I have explained in detail is absolutely typical; it is the most interesting that I have seen yet, however 'the examina- tion of matters adhering to the feet of other ducks have fur- nished me with some different Objects: some rotifers of the family Philodinadae (Arleuz, Marchais), a large number of setae from Oligocheates, one antenna of CycIOpS?, some debris of Acariens (possible parasites on the bird?), one capsule of Turbellaria? having oviform fruit, and many carapaces of a cladoceran of the genus Alona (Marchais). In the same way that the fragments of insects, these contained some diatoms and various corpuscles of which they facilitate for sure the dissemination.. The edge of the carapace isbordered with numerous thorns. The objects encountered on the tongue and on the bill have gained my attention many times by their volume. Thus I have collected in the interior of the bill of a duck some plant frag- ments attaining up to three centimeters in length and by no means desiccated. In one other case, on the edge of the tongue were found some ovoid particles of quartz being three millimeters in diameter longitudinally and about two millimeters in transverse diameter. 244 This Shows that some Molluscs, for example, of a certain Size can be transported in the same manner. I will be forced to be brief on the subject of the cultures I have still to Speak about. One of them continued for two months- from November 18, 1887, to January 17, 1888, with some material taken from the duck mentioned above, has furnished aside from other living animals, some Nematodes, and some very lively Rotifers (Philodinadae). The direct examination had not Shown these types. Some RhiZOpOds (Trinema enchelys Ehv. for example) seem to have made their appearance there as well. But one Should never rely on this experiment, the winter being an unfavorable season during which many aquatic organisms remain inactive in our climate. I dare to hope that the cultures where the presence of some living beings is certain will not delay in changing its appearance; the study of which will immediately be resumed. Nevertheless, the range of general obServationS which pre- cedes is by now obvious. They Show the important role played by the birds, and the web-footed in particular in the dissemination of organisms from fresh water. They explain the cosmopolitan char- acter Of certain types, at the same time that their presence in these isolated points and notably on some oceanic island; they explain also the introduction of these types in the basins Lacustres of recent 245 origin, or in the artificial ponds. 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