Notes: Three strains of Hypotrichomonas acosta were isolated in axenic culture. Attempts to develop a defined medium directly from a defined medium suitable for Tetrahymena pyriformis were unsuccessful. Development of partly defined media by substitution for undefined materials in Diamond's medium were more successful. Horse serum was replaced by 1 mg. % TEM-4T (a diacetyl tartaric acid ester of monoglycerides from tallow) and 0.5 mg.% cholesterol. Yeast extract was replaced by a mixture of ribonucleotides. Inclusion of several additional components permitted reducing the Trypticase concentration from 2% to 0.25%.

Notes: SYNOPSIS. The life cycles of 2 strains of Allogromia laticollaris (CSH and TPA) raised on experimentally restricted diets have been studied. Cloned cultures of the strains differed in nutritional requirements for continuous reproduction. The TPA strain was more fecund. Both strains have a basically apogamic (asexual) life cycle; only occasionally were gamonts (sexually reproducing individuals) produced. They were autogamous (self-fertilizing) and fusion of the gametes took place within the parental test.We found in CSH clones a regular alternation of vesicularly nucleated and compactly multinucleated diploid generations. The details of the life cycle of either strain are not known well enough to be able to construct life cycle diagrams more meaningful than that of Arnold. The 2 strains studied differ in cytologic and nuclear detail from each other and from the strain studied by Arnold.

Notes: SYNOPSIS. Several trichomonads from poikilotherms (Hypotrichomonas acosta, Monocercomonas colubrorum and M. sp., Trichomonas batrachorum, and Tritrichomonas augusta) grew in a nearly-defined medium to a level of 10–40% that permitted with media containing casein digests; T. augusta (4 strains) grew best. The defined medium contained TEM-4T (a semisynthetic fat based on beef-tallow fatty acids) and ascorbyl “palmitate” as sources of fatty acids. Besides ascorbyl palmitate, thiomalic and ascorbic acids, and tocopherol served as reducers and antiöxidants. High concentrations of nucleotides (adenylic, cytidylic, guanylic, and uridylic acids) seemed necessary. Indigo carmine was a useful redox indicator; the flagellates never grew in the oxidized zone. Extra trace elements and amino acids or rich natural media permitted better growth at about 34.5°C—the upper temperature limit for these trichomonads, raising the issue of a biochemical basis of a thermal barrier to the invasion of warmblooded hosts. Except for the limited carbohydrate in the nucleotides, carbohydrate could be omitted, the main substrates being a combination of glycerol, acetate, and gylcero-phosphate, posing the question of the identity of the anaerobic, presumably non-glycolytic, energy-yielding pathway.

Notes: SYNOPSIS. A polymorphic allogromiid (our strain NF) was isolated in monoxenic culture but attempts at its axenic culture failed. Growth of monoxenic cultures was stimulated by various metals and vitamins. The morphology and life cycle of this allogromiid with a prominent collar have been studied in detail. It varied in form from ovoid to elongate bioral “Shepheardella-like” forms and to irregular polyoral organisms. The organisms ranged in length from 56–385 μ (118 μ± 50.54) and in width from 35–385 μ (99 μ± 39.34). The number of nuclei per organism averaged 8.9 ± 6.3 (range 1–40). Reproduction of “Shepheardella-like” forms was by binary fission. Three types of budding have been observed and, rarely, schizogony. No evidence for sexual reproduction has thus far been seen.

Notes: Hypotrichomonas gen nov., with a feeble undulating membrane but lacking a costa, is placed in the family Monocercomonadidae Kirby of the order Trichomonadida Kirby. H. acosta (Moskowitz) from squamate reptiles is designated as the type species. It has 3 unequal anterior flagella. The proximal part of its recurrent flagellum is attached to the surface of the body along an accessory filament. A low membrane with typically a few undulations extends between the accessory filament and the body surface. The membrane, which varies greatly in length in relation to the length of the body among strains of the species, is not supported by a costa. The recurrent flagellum continues as a free posterior flagellum beyond the end of the undulating membrane and terminates in an acroneme. The capitulum of the axostyle, spatulate or scoop-shaped, is closely applied to the anteriorly located nucleus. The capitulum extends anteriorly into a crescent-shaped pelta. The relatively stout trunk of the axostyle is straight or twisted. It projects for some distance from the posterior body surface, and the terminal segment of the projecting part tapers gradually to a point. The parabasal apparatus consists of a rod- or shaped body and of a filament or filaments.In division, the full complement of flagella in each of the mastigonts is restored relatively late during division but before cytokinesis. One of the daughter individuals retains the parental undulating membrane; the other develops a new organelle. The development of the new membrane is first marked by the appearance of an acroneme, presumably recurrent flagellum, early in division. The proximal part of this flagellum becomes progressively attached to the body surface. The process of attachment is paralleled by differentiation of the accessory filament and is followed by the appearance of the undulating membrane. The parental axostyle is resorbed during division, and each daughter individual develops this organelle de wove. In the very early stages of division the parabasal consisted of two arms and in all more advanced stages of a single rod-shaped body applied to a single filament.The significance of Hypotrichonionas in the evolution of trichomonads is discussed. Its morphology and morphogenesis suggest that this genus may represent an important step in the evolution of Trichomonadidae from Monocercomonadidae.

Notes: SYNOPSIS. Two new dinoflagellates, isolated from Long Island Sound and maintained axenically, are described. Gyrodinium instriatum n. sp. changes shape and size as cultures age. Glenodinium halli n. sp. is a delicately-armored form whose plate structure differs slightly from the present diagnosis of the genus, but not sufficiently like any other genus in morphology or life history for inclusion elsewhere. In G. halli the size of the motile form and predominance of motile or vegetative stages are nutrient dependent. Both dinoflagellates illustrate the need for morphological descriptions based upon continuous culture methods in defined media.

Notes: Summary Monhystera denticulata Timm, a free-living nematode present in the aufwuchs assemblages of several marine macrophytes located in North Sea Harbor, Southampton, New York, was isolated from Zostera marina and established in laboratory culture in order to study the influences of temperature and salinity on its life history. Under experimental conditions, M. denticulata has a generation time (Measured as the time elapsing between the first egg depositions of consecutive generations) of 10–12 days at 25° C and 26‰ S, which represent optimal growth conditions in the laboratory. The organism has a generation time of 20 days at 25° C and 13‰, 17 days at 25° C and 39‰, 18 days at 15° C and 26‰, 36 days at 15° C and 13‰ and 34 days at 15° C and 39‰. As conditions vary from the optimum of 25° C and 26‰ S, a decrease in temperature of 10° C and an increase or decrease in salinity of 13‰ results in a doubling of the generation time. At 5° C the generation time is about 180–197 days. Assuming optimum conditions and average generation time, about 15 generations of M. denticulata could occur in North Sea Harbor during the year. The number of generations occurring in reality is probably less, however, due to the fact that the females deposit their eggs over a period of several days.

Notes: Summary Chromadora macrolaimoides Steiner, a free-living nematode present in the aufwuchs assemblages of several marine macrophytes located in North Sea Harbor, Southampton, New York, was isolated from Enteromorpha intestinalis and established in laboratory culture, where its life history and feeding habits were studied. Under the experimental conditions (25 C and 26‰ S) the worm has an average generation time (22 days) and average life span (45 days) similar to other chromadorids which have been studied in the laboratory. Tracer-feeding experiment with 32P-labelled bacteria, diatoms and chlorophytes indicate selectivity by the worm in both the ingestion and apparent digestion of potential food organisms, with the diatoms and chlorophytes being the preferred foods. Out of a total of 20 species of algae and 14 species of bacteria, two species of diatoms (Nitzschia acicularis and Cylindrotheca closterium) were found which are capable of sustaining indefinite growth. Bacteria-free culture has not been established, however, due to the extreme sensitivity of the worm to antibioties. A comparison of the feeding habits of C. macrolaimoides with Rhabditis marina, another marine nematode fed the same potential food organisms is made, and the influence of selective feeding on the spatial and temporal distribution of marine nematodes is discussed.