ALBERT

Notes: Vorticellas were maintained on Bacillus cereus in a medium composed of unbuffered Proteose-Peptone, Cero-phyl, and the filtrate from boiled wheat kernels. Vorticella-free aliquots of culture medium were analyzed at critical intervals, beginning immediately before encystment and until re-encystment. Control aliquots changed insignificantly; significant changes were detected in the experimental cultures. Osmolality increased from 14 to 18 milliosmols/1, Na+ stayed in the range 2 to 3 milliequivalents/1, K+ decreased slightly below the initial value of 5 mEq/1, Ca++ decreased from 5.7 to 4 mg.%, CI fluctuated slightly down and up from 112 mg.% during the initial 8 hr., then down to 〈 104 mg.%, phosphate decreased steeply from 14 to 〈 0.6 mg.%, pH increased from 6.7 to 8.5.The results indicate that suboptimal levels of K+ Ca++ and phosphate impose upon V. microstoma a physiological stress which leads to encystment. The role played by Na+ and CI- was not clear. Osmolality was a major factor in excystment but not in encystment.

Notes: SYNOPSIS. Vorticella microstoma was grown non-axenically and axenically at pH 6.4. Vorticellas were maintained indefinitely on Bacillus cereus in a medium composed of Proteose-Peptone, Cerophyl, and the filtrate from boiled wheat kernels. Prolific growth occurred in 2-membered cultures. A medium containing hydrolyzed gelatin, aqueous liver extract, yeast nucleic acid hydrolysate, glucose, and penicillin is recommended for axenic growth.The potential value of vorticellids as research tools is discussed together with metabolic implications of supplementing sterile Proteose-Peptone broth with natural substances in particle form. The ineffectiveness of adding tryptophan, thiamine, glycine, and chelating agents to the axenic medium was considered. Refinements of the axenic medium are on trial.

Notes: SYNOPSIS. Pyxicola nolandi, n. sp., is a loricate peritrich favored by clean pond water. The yellowish-brown lorica measures 74 × 25 μ, on the average; supported by a noncontractile stalk, it resembles a vase with a slightly curved neck bearing an oblique opening. The stalk, ∼ 13 × 6 μ, is marked by vertical lines which give it a corrugated appearance. The bluish-gray translucent truncated body is protrusible beyond the lorica's opening, averages 87 × 15 μ, and is characterized as follows. An expansive sphincter collar that extends into a narrow shelf; 3 rows of concentrically spiralling peristomial membranes; prominent “soie de Lachmann” visible when peristomial region expands; a slightly lobulated discshaped operculum originating on the ventro-lateral surface of the peristomial collar, averaging 15 μ in diameter, and overhanging the edge of the lorica when the body extends; one contractile vacuole; C-shaped macronucleus; micronuclcus ∼ 6 times 3 μLarge populations were found in June, July, and August, attached to the alga Rhizoclonium hieroglyphicum and to other kinds of filamentous algae. The peritrich was scarce in winter, but it was found even when the pond was covered with ice, providing that Rhizoclonizm survived in the cold weather.Some aspects of binary fission, lorica formation, conjugation, environmental pREFERENCES, and maintenance are described. The peritrich was cultivated in laboratory aquaria. Clones were established repeatedly from pond D-7 and from laboratory aquaria. Loricae were stained successfully with Congo red. Nuclear phenomena were revealed with the Feulgen technique. Silver impregnation was unsuccessful, thus far.

Notes: SUMMARY. We have been investigating asexual and sexual stages in the life cycle of a ciliated protozoan, Vorticella microstoma, with the object of determining characteristic patterns at the biochemical level. In the course of our studies we have made progress in the chromatographic analysis of acid hydrolysates of the asexual (neuter) and the sexual forms. Our findings demonstrate that it is possible to obtain from the neuter stages ninhydrin-positive patterns which are characteristically different from the patterns derived from the sexual stage. These patterns are believed to be indicative of the biochemical peculiarities of the organisms. Ninhydrin-positive patterns of the sexual stages of Vorticella microstoma are composed of 15 bands, in contrast with 10 bands observed in the patterns of asexual stages. In regard to color, bands 1–10 in the neuters are similar to bands 1–10 in the sexuals. The difference between the two ninhydrin-positive patterns is believed to be significant, in that it indicates the biochemical peculiarities of the respective kinds of Vorticellae. It is suggested that chromatographic analyses may lead to the identification of the substance or substances liberated by the macroconjugant for the purpose of attracting microconjugants. The methods which have been successfully used in this study may be helpful to investigators who utilize protozoa as experimental organisms. We believe the methods and results reported here may have applications in the genetics of protozoa and in biological studies devoted to metazoan subjects.

Notes: SYNOPSIS. Cultures of Telotrochidium henneguyi, begun with logarithmic phase cells, were employed in an effort to produce synchronized fission by heat treatment. The cells tolerated a temperature range of 20–50 C; temperatures above 50 were lethal. When cells were exposed to a single shock for 30 min, 30–40 produced 0–50% encystment with total excystment after 10 min exposure to room temperature (heat shock range). No encystment occurred between 20–30 (intershock range). Encystment and excystment time varied directly with temperature between 40–50.The most effective procedure for inducing synchronized fission consisted of 6 cycle program of 38/28 C (shock temperature/intershock temperature) administered for 15/15 (shock/intershock duration in min). Division indices (DI = cells dividing/total population X 100 =%) ranged from 12–66% with a mean of 37.25%. In control cells, division indices ranged from 2–20% with an average of 12%. Inferences from these independently derived findings are discussed.

Notes: SYNOPSIS. Telotrochidium henneguyi was cultured axenically. The major nutrients in medium T-3 were liver extract (1:20 National Biochemicals Co.), hydrolyzed yeast nucleic acid, glucose, and dl-β-hydroxybutyric acid; these were fortified with phosphate buffer, EDTA, and penicillin. The supplements were: 18 amino acids, 7 vitamins, 10 salts supplying trace-metals; uridylic, cytidylic, guanylic, and adenylic acids; thymidine-5-diphosphate, nicotinamide mononucleotide, and choline. Optimum conditions for axenic cultivation were obtained with phosphate buffer 2 × 10−1M, penicillin 5,000 U.S.P. units/ml, 23°C, pH 6.8. A monoxenic maintenance medium (“T-broth”) allowed prolific growth and produced trxmendous populations. It was composed of Bacillus cereus in an aqueous broth-concoction of Proteose-peptone, Cerophyl, and wheat kernels. Axenic T-3 medium supported serial subculture; yields of peritrichs were comparable to those in T-broth. Axenic yields were poorer in medium lacking the T-3 supplemenits but containing the major nutrients fortified with acid-hydrolyzed gelatin, serine, riboflavin, EDTA, CaCL, FeCI3, KCI, MgSO4·7 H2O, phosphate buffer, and penicillin. For rapid axenination, excystment was induced by vibrating encysted peritrichs ∼ 30 sec in a Vortex Jr. mixer. Freshly excysted animals were washed by centrifugation. A salt solution, not distilled water, was used for washing inoculants. Inocula consisting of 1 × 103 or more animals were obtained by conventional centrifuge methods.Extension of this investigation to construction of a chemically defined medium is discussed.

Notes: SYNOPSIS. A study of the ectoplasm and infraciliature of Spirostomum ambiguum has been made with light and electron microscopy. Proceeding from the exterior, the cortex is described as the highly structured ectoplasmic portion of Spirostomum consisting of ectoplasmic ridge components and ectoplasmic furrow components. The pellicle invests the ectoplasmic ridges, ectoplasmic furrows, cilia, and adoral membranelles. Electron micrographs show that the cell surface is bounded by two membranes, for the most part parallel. The outer one is continuous with the unit membrane of each cilium; the course of the inner membrane has not been determined in areas bearing cilia.The ridge components are: (i) a collection of fibrils called peripheral ectomyonemes, situated beneath the inner membrane of the cell surface, oriented parallel with the organism's longitudinal axis and following the peripheral contours of each ridge; (ii) distinct rows of tubular fibrils which form a longitudinal bundle called lateral ectomyonemes, situated in the lower portion of each ridge and oriented parallel to its spiral course; (iii) cytoplasmic matrix suspending and surrounding components (i) and (ii). Mitochondria and unidentified electron-opaque oval-shaped or round bodies called X-bodies are dispersed in the ectoplasmic ridges but they also occur in the endoplasm; therefore, they are considered to be transient inclusions instead of constituents of the ridges.The furrow components are: (i) the body cilia and adoral membranelles; (ii) the kinetosomes; (iii) root fibrils, called kinetodesmal strands, of the adoral membranelles; (iv) cytoplasmic matrix suspending and surrounding the furrow components.A trabecular band of randomly oriented filaments, called the endomyoneme is described as possibly the structure called M-band by Randall and Jackson, 1958, and contractile fibrillar system by Yagiu and Shigenaka, 1963.It is suggested that collections of distinct fibrils in Spirostomum be called fibrillar complexes. Acceptance of this suggestion will lead to the designation of four distinct fibrillar complexes in heterotrichs; all four complexes have been described or mentioned in previous studies.Some findings of previous investigators are discussed and confirmed (especially pertaining to Randall and Jackson, also Yagiu and Shigenaka, cited above). Inferences from these independently derived findings are discussed.

Notes: SYNOPSIS. The peritrichs have taught naturalists, biologists, zoologists, and other kinds of scientists many of their secrets. Consequently, we know many details regarding their gross structure, ultrastructure, ecology, and life history. Their sexual phenomena are rather clearly understood. While general agreement has been attained as regards the forementioned topics, the taxonomic position of peritrichs is still vigorously debated. In this address representative examples of advances in our knowledge of peritrichs will be reviewed … representative instead of all-inclusive examples. A plea for the restoration of peritrichs to a taxonomic position above spirotrichs will be made. The Society's members will be assured that all protozoologists will have the opportunity to help design a completely International Commission on Protozoology in the International Union of Biological Sciences and they will be assured that such a Commission can and should be planned at the 1969 International Conference in Leningrad.