ALBERT

Notes: SYNOPSIS. Ten new species of colorless Eugleninae are described. One of them is the type of a new genus (Calycimonas physaloides nov. gen. nov. sp.). Especially remarkable forms are: Astasia acus, a species related to the green Euglena acus but without chromatophores and stigma and considerably smaller; Astasia edax, a species with animal-like nutrition and peculiar ecological adaptations; Petalomonas striata, a form which is not spirally but transversely striated; and finally Hyalophacus caecus, which was observed already by Klebs and Pochmann but was not sufficiently described. The new genus Calycimonas (ex fam. Peranemacearum) is related to Tropidoscyphus on the one hand (nearly the same shape as e.g. Tr. octocostatus, but possessing only a single flagellum) and to Petalomonas on the other hand (mode of nutrition and of swimming, formation of the reservoir, rigidity: however in transverse section not flattened).The Astasiae with zootrophic nutrition are collected in a new subgenus Astasiae devorantes or Phytophaga which comprises besides A. edax three species already described by Skuja.

Notes: SYNOPSIS. H. canis (James) is reported from three of 36 stray dogs at Singapore and one of 34 palm civets (Paradoxurus hermaphroditus Pallas) from Jeram, Selangor. H. muris (Balfour) was found at Singapore in 10 of 61 examples of Rattus norvegicus (Erxleben) and five of 23 of R. rattus diardi (Jentinck), and is also recorded from six of 139 examples of R. r. jarak (Bonhote) from Pulau Jarak, Straits of Malacca. Neutrophils of P. hermaphroditus containing gametocytes of H. canis exhibit pronounced karyorrhexis, while most of the parasites themselves show signs of pyknosis.

Notes: SYNOPSIS. A total of 155 clones of Stylonychia putrina collected in nature were tested for mating type. Two sexually isolated varieties were found among them: Variety I with 12 mating types and Var. II with 11. Five of the clones isolated in the laboratory belonged to 3 additional mating types of Var. I making a total of 15 mating types in this variety.Crosses were made among 6 of the mating types of Var. I, and 114 clones of progeny were raised and tested for mating type. No simple genetic explanation of the results was evident.An abnormal type of conjugation with fusion of the conjugants and subsequent fission was studied. Both parental mating types sometimes segregated during the first two fissions of a fused animal. Mating type determination in the progeny of the fused animals seemed to parallel that of the progeny of the normal conjugants in that in both cases certain mating types tended to be dominant over others.

Notes: SYNOPSIS. The cytochemical procedure of Sen was used to demonstrate urease activity in Tetrahymena pyriformis S.

Notes: SYNOPSIS. The chimpanzee is shown to behave like man to infection with Babesia divergens or bovis: the intact animal is totally resistant, whereas the splenectomized animal develops a fulminating infection accompanied by blackwater. The splenectomized rhesus monkey reacts in the same way also, but splenectomized rabbits are insusceptible. In the chimpanzee the typical accolé position of the “divergens” organisms (as seen in cattle) is absent, but occurs in the rhesus. It is suggested that latent piroplasmosis in man may exist on a large scale in rural populations in infected localities.

Notes: SYNOPSIS. Studies cf 3 conjugating strains of Tetrahymena pyriformis grown in a bacterized medium indicated the presence of a system of alternative immobilizing surface antigens. Each strain was found to have the potential for the expression of 3 serologically unrelated antigens. These consisted of a “high temperature” antigen (expressed in the range of 20–35d̀C), a “low temperature” antigen (exhibited in cultures at 10d̀), and an antigen induced by growth in the presence of the above-mentioned “high temperature” antiserum. Normally, by the immobilization reaction, only one of these antigens could be detected as present on any one organism at any given time. The “high temperature” antigens of two of the strains were serclogically related while that of the 3rd strain did not cross-react with antisera to the other two. The “low temperature” antigens of the 3 strains were serologically related as were the antigens induced by growth in antiserum.Studies of inbred hybrids of two of the strains indicated a potential for still more alternative antigens as well as for the spontaneous expression of the antigen which in the parental strains could only be induced to appear by the presence of the specific antiserum.Study of the 3 strains in axenic medium indicated the existence of a more complex system and the possibility that two or more immobilizing antigens may be present simultaneously.

Notes: SYNOPSIS. The establishment of Eimeria tenella sporozoites in the cecal mucosa of the chicken is described. The invasion process was similar to that reported for E. necatrix by Van Doorninck and Becker. Sporozoites were found to pass through the surface epithelium of the cecal mucosa into the lamina propria. Within the lamina propria the sporozoites were engulfed by macrophages and transported to the cells of the glands of Lieberkühn. Development of the sporozoites ensues within the gland epithelium.

Notes: SYNOPSIS. The morphology of the vegetative phase and the sexual process in Pandorina morum have been studied in detail under controlled laboratory conditions. A number of strains from various areas of the United States, all essentially indistinguishable on morphological grounds, were analyzed for sexual compatibility. Forty-seven heterothallic clones were found to represent 15 separate pairs of mating types, or a minimum of 15 syngens (sensu Sonneborn, 1957). Two clones proved to be homothallic. Nine additional clones, which were not observed to mate with any strain, can be classified only after further collecting. The separate heterothallic pairs of mating types are sexually isolated by factors acting at the stage of gamete production; incompatible mating types produce no gametes when they are mixed together. The geographical distribution of the sets of mating types is very incompletely defined as yet, but they are not strictly endemic forms. Thirty heterothallic strains, representing each of the 15 sexually isolated sets of mating types of Pandorina morum, have been deposited with the Culture Collection of Algae maintained at Indiana University, Bloomington, Indiana, U.S.A.

Notes: SYNOPSIS. When grown in CPLM medium or in a similar medium containing glucose instead of maltose as its sugar, Tritrichomonas foetus, Strain O, was much more sensitive to injury when frozen to -21d̀ C. in the presence. of 1 m glycerol during the initial and logarithmic phases of its population growth curve than at its peak and for some time thereafter. In 7 experiments in which the population peak occurred an average of 28.1 hours after inoculation, the average culture age at which the protozoa first survived freezing was 20.3 hours, at which time the population had reached 52.6% of its peak. The optimum culture age for survival after 1 day of freezing averaged 37.7 hours at which time the population averaged 75.6% of its peak. The optimum culture age for survival after 7 to 15 days of freezing averaged 32.3 hours, at which time the population averaged 82.7% of its peak. Better survival upon freezing was obtained in those experiments in which the population peak was reached relatively rapidly.

Notes: SYNOPSIS. Specimens of Blepharisma undulans were transferred from young clone cultures to a rotocompressor with 3 mm3 of culture medium. The organisms were slightly compressed and photographed at various intervals with dark field illumination. The sequence of macronuclear and cytoplasmic changes was compared with similarly followed Feulgen preparations.The cycle falls into several phases: (1) an interphase (12–24 hr.) in which the organism increases in size while cytoplasmic and macronuclear appearances remain unchanged. During this phase, the macronucleus consists of 3 to 5 nodes of various sizes connected by strands. (2) a pre-condensation phase (1 hr.) in which a new posterior peristome and cytopyge appear without visible macronuclear change. (3) a condensation phase (10–20 min.) in which the macronuclear nodes coalesce into a round mass without dissolution of the central nodes or strands. (4) a postcondensation phase (1 hr.) characterized by: (a) elongation of the condensed macronucleus into a rod-like shape followed by typical nodal formation; or (b) elongation of the condensed macronucleus into a form resembling the letter “J”, followed by nodal formation, and resulting in a double row of nodes in one daughter and a single row in the other. Separation occurs at this time, initiating the new interphase. The development of the j-form macronucleus. the lack of obliteration of the central nodes, and the characteristic interphase condition distinguish this strain of B. undulans from others described elsewhere.

Notes: SYNOPSIS. Six species of astomatous Infusoria, 4 new, are described from the alimentary tract of Oligochaeta from Ochrida Lake. Two belong to Ochridanus, characterized by a cytoskeleton whose “V”-shaped basal piece bears on its branches two articulated hooks brought together by a skeletal blade. This genus found in Ochridanous Tubificidae represents the counterpart of Anthonyella in the Lumbriculidae of Lake Baïkal. The presence of Ochridanus in these worms concurs with the almost complete absence of representatives of Radiophrya of which we have found but one species. A species of Metaradiophrya, a genus unknown to date from the Lumbricidae, was noticed in Glossoscolecidae. The different species of Ochridanus, Anthonyella, Metaradiophrya, and Radiophrya compose a very homogeneous group of Radiophryinae. The study of the ciliary rows and the cytoskeleton of 2 species of Juxtaradiophrya, parasitic in the Lumbriculidae of Ochrida, shows that, in the morphology of these ciliates, as well as in the same forms from Lake Baïkal, many transition characteristics exist between the Radiophryinae and Hoplitophryinae or Mesnilellinae. These 3 sub-families, with their undeniable genetic kinship give coherence to the family Hoplitophryidae.

Notes: SYNOPSIS. Histochemical techniques were used to demonstrate the intracellular distribution of some hydrases, hydrolases, oxidases, and dehydrogenases in Stylonychia pustulata. The hydrase, aconitase, was confined to the mitochondria. Zymohexase activity occurred in the cytoplasm and probably in the mitochondria. The hydrolases, acid and alkaline phosphatases, lipase, and urease, were localized in the mitochondria. Lactic and glutamic dehydrogenases were confined to the mitochondria. Peroxidase and glycerophosphate dehydrogenase were absent.

Notes: SYNOPSIS. When two strains cf T. pyriformis that do not require exogenous pyridoxine are crossed, all progeny grow without the vitamin. Offspring from crosses of two pyridoxine requiring clones require pyridoxine with the exception of a few which will grow without pyridoxine. The ratio is approximately 3:1 favoring the pyridoxine requiring category. In matings involving the homozygous dominant pyridoxine requiring clones with the double recessive mutant, that is +/+ X p/p, all of the resulting progeny need pyridoxine. Test crossing these heterozygotes (+/p) with the parental pyridoxine non-requiring clones (p/p) gives offspring approximating a 1:1 ratio. Matings between two heterozygotes derived from breeding experiments also yield progeny in approximately 3 pyridoxine requiring: 1 pyridoxine non-requiring. All data indicate selection for the heterozygote in the population and a possible selection against either homozygote. The great abundance of heterozygotes and rarity of recessive homozygotes in natural habitats corroborates these findings. The genetic evidence supports a single gene hypothesis although the possibility of multiple closely linked genes cannot be ignored. There is also the possibility that a dominant suppressor gene may function in blocking the activity of the pyridoxine mutant genes. Moreover, if this gene exists it may be incompletely dominant since the heterozygote grows slightly on deficient media.

Notes: SYNOPSIS. Trichodina urinicola was found in newts, Triturus cristatus and T. taeniatus, in three localities in Czechoslovakia. The ciliate populations showed important differences on the basis of which they were separated as three new forms of this species: T. urinicola f. typica, T. urinicola f. bohemica, both from Triturus cristatus, and T. urinicola f. taeniatus from Triturus taeniatus. The great variability of trichodinids is evident from the literature as well as from our own observations, so that these new forms are to be regarded as provisional ones until it is possible to decide on the basis of a large number of observations the extent of specificity and variation of individual endozoic species of Trichodina. A detailed description of these forms is given as well as a comparison with the known species of trichodinids inhabiting the urinary tract of amphibians. A brief comment on the present taxonomy of the Urceolaridae in general is outlined.The need for a uniform description of these ciliates is emphasized; in connection with this, the taxonomic value of individual body characters is discussed. Special attention is paid to the adhesive disk of Trichodina, the structure of which is of greatest importance in the taxonomy of this group. On the basis of Dogel's and Fauré-Fremiet's descriptive methods employed in study of trichodinids, a proposal of a uniform description of Trichodina is made which involves all the important features of these protozoa.

Notes: SYNOPSIS. Several substrains of Euglena gracilis var. bacillaris made chlorotic by treatment with pyribenzamine or streptomycin, or by growth at high temperature (35–36°C.), have been examined for their carotenoid content. They differ from the normal green strain both qualitatively and quantitatively. Some strains produce no detectable carotenoids while the carotenoid concentration in the strains producing most is at best only one-fifth that of the normal strain. In all substrains producing carotenoids, the carotene fraction consists of β-carotene accompanied by some members of the phytofluene series. In only two of these substrains, HB-G and PBZ-G3, are xanthophylls produced in significant amounts. In HB-G, the main pigment is echinenone, and in PBZ-G3 it is zeaxanthin. The significance of these findings is briefly discussed.

Notes: SYNOPSIS. Association of gamonts of Pyxinia crystalligera takes place in the midgut of its beetle host, Dermestes vulpinus. At 25°C. the development of gametocysts to the point of liberation of sporocysts is completed between about 15 hours and 27 hours after the gametocysts are deposited with fecal material. Dehiscence is favored by relative humidities of 0% to 90%, but is not favored by a relative humidity of 100%. During the early development of the gametocysts outside the host, the crystals and paraglycogen granules in the cytoplasm of the associated gamonts become concentrated in large masses. The gametes are formed at the periphery of the gamonts. After fusion of the gametes takes place and the sporoblasts begin to develop, the residual cytoplasm containing the inclusions moves outward to form a continuous layer next to the gametocyst envelope, so that the sporoblasts become crowded into a central core. A few hours before dehiscence is initiated a clear area appears on the upper side of the gametocyst. The contents of the gametocyst begin to shrink away from the envelope except in the region of the clear area. Eventually the sporocysts emerge through the clear area and press against the envelope of the gametocyst, causing formation of a conical papilla in the envelope. With continued pressure from the sporocysts, the papilla ruptures at its tip, and the sporocysts emerge in a continuous thread until dehiscence is completed. The thread of sporocysts may attain a length of about 11 mm.

Notes: SYNOPSIS. The steps leading to purine ring closure were examined in Strigomonas oncopelti. The flagellate has an obligate adenine requirement (hypoxanthine and guanine are inert) when grown without p-aminobenzoic acid. The imidazole counterpart of adenine but not the imidazole counterpart of hypoxanthine was active. A pathway for purine biosynthesis compatible with these results is sketched.

Notes: SYNOPSIS. The Nuttallia of the gerbil is transmitted by Rhipicephalus secundus only if the infected meal is taken by the larva, and the subsequent nymphal stage is the only one which is able to infect. There is no transovarial transmission.Infective trophozoites remain in the larval caeca for about 12 hours after gorging. The infective nymph is able to give rise to new infections at various times after it has been allowed to feed, but never later than the third day after disengagement from the host.

Notes: SYNOPSIS. The process of autogamy in unassociated individuals of Paramecium polycaryum was reported by the author in 1954. In May, 1955, conjugation was first seen in this species in cultures collected by me at Annamalainagar, South India, thus removing it from the list of non-conjugating species. This appears to be the first instance in which the process of autogamy was detected prior to observation of conjugation in the same species. Autogamy occurs in singles of the Indian race and appears to be similar, cytologically, to that of American races. The details of the micronuclear behavior in conjugation parallel those of autogamy in singles. In fact, the conjugation process seems to be one of double autogamy (cytogamy), rather than of reciprocal gametic interchange. Paroral cones, often of fair size, are formed but breakdown of the cones to permit micronuclear passage has not been observed. In conjugation there are the usual three pregamic divisions; the first shows four characteristic crescents. The resulting nuclei may all participate in the second division. Fertilization occurs in the paroral cone area. Frequently, separation of the conjugants takes place immediately after the first division of the synkaryon. The old macronucleus undergoes very little change prior to the last postzygotic micronuclear division in the ex-conjugant, when it goes into a skein condition. Four macronuclear and four micronuclear anlagen are formed in the ex-conjugants at the completion of reorganization. On occasion giant individuals of P. polycaryum were observed to have ingested numbers of Tetrahymena pyriformis. The presence of an unidentified rod-like organism in the cytoplasm of the paramecia (non-conjugating) was detected in one collection from Bangalore, India.

Notes: SUMMARY. The mating behavior of 16 stocks of Paramecium multimicronucleatum from 12 states has been studied. Mating always follows a decline in nutritive conditions. The evidence indicates that there exists one set of four interbreeding mating types. The period of subculture strongly influences the mating reactions, many stocks mating only long after isolation from nature and culture in the laboratory under the restricted conditions employed. Selfing was observed to occur in many races, usually after long periods of subculture. It first occurred in only a small proportion of the population, later in a larger proportion. The similarities and differences between P. multimicronucleatum and other species of protozoa showing multiple mating types are discussed.

Notes: SUMMARY. A study of the temperature-pressure relationship in oxygen poisoning of Paramecium caudatum was undertaken, as the initial step in defining some of the major factors in in vivo oxygen poisoning. Paramecium was selected because it was relatively simple to culture in a pureline clone, large numbers were readily obtained, and it was large enough to be clearly visible under low magnification. The protozoa were exposed to oxygen pressures of 0 (100%), 15, 30, 45, 60, 75, 90, 105, 120 pounds/inch2 gauge pressure at each of the following temperatures: 1, 5, 10, 15, 20, and 27°C. Exposure was accomplished in a transparent, high-pressure lucite tank which permitted visual observation with aid of a dissecting microscope. It was found at temperatures above 5°C. that oxygen toxicity varied directly with pressure, but below this temperature, with oxygen tensions of 1–2 atmospheres (absolute), oxygen toxicity varied inversely as the temperature. The possibility is advanced that oxygen may be affecting two cellular processes (perhaps enzymatic), one of which is temperature-limited below 5°C. and would, therefore, decrease the death time as the temperature is decreased. Several experiments performed at 3°C. produced a death time intermediate between results obtained at 1° and 5°C.

Notes: SUMMARY. A stock of Paramecium bursaria is described in which the peniculus contains only 8 columns of cilia and associated “granules”. The gullet system is composed of organelle complexes almost exclusively, and incorporates no underlying or transecting extensions of “pellicular fibrillar systems”. A brief discussion is presented on the value of the peniculus as a taxonomic criterion in separating “aurelia” and “bursaria” groups in Paramecium.

Notes: SUMMARY. Strains E, S and W of Tetrahymena pyriformis were examined for their ability to carry out the reactions of the Krebs-Henseleit urea cycle, using growth and enzyme studies. None of the strains was able to grow on either citruliine or ornithine in place of arginine, and proline was as active as citrulline or ornithine in sparing arginine. So little citrulline or arginine was synthesized by cell-free preparations as to be of no significance in the growth or nitrogen metabolism of the ciliates. Slight arginase activity could be detected in homogenates, but no urea was found in cultures. No urease activity could be detected using urea-C14.

Notes: SUMMARY. An eimerian coccidian is described from the flying squirrel, Glaucomys volans, in Florida. It is identified as the same eimerian described by Roudabush from the flying squirrel in Iowa as Eimeria sciurorum. Evidence is presented that Roudabush incorrectly identified the organism. It is renamed as E. parasciurorum nov. sp. Mature oocysts have mean measurements of 29 × 16°, an index of 1.82, are cylindrical with rounded ends, have a dual membrane, and no extra residual body. Oocysts are without micropyle. Four egg-shaped, mature sporocysts in the oocyst have mean measurements of 11.2 × 6.2°, an index of 1.81, contain an oval, granular, intraresidual body and two pyriform sporozoites 10 × 3.2°, index 3.11.

Notes: SUMMARY. Two centrioles, an old one and a new one, are always present in the resting cell. From prophase onward two new ones and two old ones are present. Beginning with the resting stage, five types of centriole life cycles are described and compared with one another: In type 1, both centrioles are elongate; in type 2, the old one is long and the new one, which is short, elongates in prophase; in type 3, both are short, both elongate in prophase, and both, except for their anterior tips, degenerate in late telophase; in type 4, both are long but in prophase their distal ends become free of the rest of the centrioles, these ends migrate to center or posterior end of cell, where, after they produce the achromatic figure and it completes its function in nuclear division, they degenerate; in type 5, both are short and neither elongates at any stage of its life cycle.New centrioles are produced by the anterior ends of old ones. In their first generation, centrioles produce only extranuclear organelles (flagella, parabasals, axostyles, etc.); in their second and later generations, they produce only the achromatic figure (gametogenesis in Trichonympha and reorganization in Barbulanympha and Rhynchonympha are exceptions to this rule).The distal ends of centrioles in some types of cycles are surrounded by centrosomes; in others they are not. In one type of centriole life cycle a small central spindle is present in the resting cell in two genera; in the other types this is not the case.

Notes: SUMMARY. The endogenous development of the life cycle of Eimeria alabamensis Christensen, 1941, occurs in the nucleus of the intestinal cells of cattle. Calves were killed at various intervals after inoculation with infective oöcysts to study the endogenous cycle. Excysted sporozoites were found in the contents or scrapings from the walls of rumen, omasum, small intestine, cecum, and colon. They were found in the cytoplasm of intestinal epithelium at 2 days. Schizonts were found in the nuclei beginning at 2 days, but the number was low by the 8th day. Merozoite numbers usually ranged between 16 and 32. Some host nuclei contained as many as 48 or more, but these appeared to be the result of more than one schizont merging in the same host nucleus. Merozoites were slender, spindle-shaped bodies while still in the schizont walls, but were short with bluntly rounded tips when found in intracellular spaces and crypts. Gametocytes were found as early as the 4th day. Most of the stages of gametogenesis were limited to the lowest third of the small intestine, but in heavy infections some were also found in the cecum and upper colon. Microgametocytes were multinucleate and were more densely stained than the uninucleate macrogametocytes. The ratio of macrogametocytes to microgametocytes in 100 gametes was 78: 22. Oöcysis with “shells” were found in sections of the lower 20 feet of the ileum on the 6th day, which coincided with the shortest prepatent period reported previously. As many as three schizonts or microgametocytes or four or five macrogametocytes or oöcysts could be found in the same host nucleus. The variations in shape of the oöcysts appeared to be dependent on the number of oöcysts crowded into each nucleus.

Notes: SUMMARY. The role of centrioles in achromatic figure production is considered when the number present varies from 1–8. Each centriole after it becomes elongate produces astral rays from its distal end. Some of these rays remain free; some, by joining centromeres, become chromosomal fibers; and some, by joining and growing along those produced by one or more other centrioles, produce the central spindle portion of the achromatic figures. Thus, one centriole may function cooperatively with one to several others in the production of central spindles. But at least two centrioles must be present, and in the proper spacial relation to each other, to form a central spindle; one by itself can form only free astral rays, no central spindle or chromosomal fibers.The flagellated areas (to which the centrioles are anchored anteriorly) play an important role in determining the position of the distal ends of the centrioles with respect to one another, and the position of these ends, in turn, in a large measure, determines the types of achromatic figures produced, particularly the number of central spindles.

Notes: SUMMARY. Observations on binary fission of Lacrymaria olor show that it is a transverse fission. It involves probable intranuclear division of the micronucleus. Both micro- and macronucleus elongate in preliminary stages. Each is ultimately divided as cytoplasmic constriction cuts the spindle fibers of the former, and the connecting, nucleoplasmic thread of the latter.Surging movements of cytoplasm after fission elongate the daughter organisms and move new nuclei to normal, central sites. The anterior proboscis of the posterior daughter regenerates suddenly, complete with coronal cilia. Metachronal waves along ciliary meridians, strongly reversed on the posterior daughter, cause an oscillating movement which pulls the two apart, except for a slender, pellicular thread, ultimately severed. Until broken, this thread connects the rear tip of the anterior animal to the forward end of the proboscis of the posterior one. The organism is semi-quiescent, with proboscis retracted (except spasmodically) throughout fission. After fission the anterior animal quickly begins feeding movements and soon swims away. The posterior animal requires about half an hour before being able to begin feeding and swimming movements.The anterior contractile vacuole of the original animal becomes the primarily active vacuole of the anterior daughter; the posterior one that of the posterior daughter. Missing vacuoles are regenerated by the daughters in about one hour after fission. The division process requires about one hour for completion at 22.4°C.

Notes: SUMMARY. The multiplication rate of Tetrahymena pyriformis HS in proteose peptone medium was measured at 12 temperatures between 18.4°C. and 36.6°C. At the temperature optimum, 32.5°C., the generation time is 2.25 hours. The upper lethal temperature lies between 36.6°C. and 38.0°C. Similarly, a study of Tetrahymena pyriformis GL revealed a temperature optimum for multiplication of 29°C. with a generation time of 3.70 hours. The upper lethal temperature falls between 34.6°C. and 35.4°C. At all temperatures employed the HS strain of organisms multiplies more rapidly than strain GL. Under identical conditions, the two strains have distinctly different growth optima, upper lethal temperatures and growth rates.As measured by multiplication rate the readjustment to a sudden change in temperature (from 18.4°C. to 27.7°C.) is completed very rapidly, with an effective lag time of about 1 hour. Such a shift in temperature gives rise to a small degree of division synchrony during the first and second population doublings which follow. Subsequently, all traces of division synchrony are lost.

Notes: SUMMARY. Test tube cultures of Tetrahymena pyriformis GL in the early maximum stationary phase of growth were used as inocula. After the elapse of a short lag phase a rapid transition to the logarithmic phase of growth was observed in cultures grown on peptone media(4,6). During the early phase of exponential multiplication more cells (as expressed on a percentual basis of the population number) were in the visible stage of cytoplasmic fission than after the elapse of 5 to 6 generations. Analysis of comprehensive data suggests that the higher division index in the early logarithmic phase of growth is not the expression of a synchronization of cells in metabolic respects but rather indicates a prolongation of the stage of cell fission during this phase.

Notes: SUMMARY. Ochromonas malhamensis (Pringsheim strain) can be grown above 35.5°C.; below 35°, the previous chemically defined medium supports dense growth. The B12 and thiamine requirements rise steeply with temperature, and growth promotion by folic acid emerges; folic acid spares the enhanced B12 requirement. B12 is spared also, perhaps wholly bypassed, by purines + pyrimidines + amino acids (below 35°, exogenous purines, pyrimidines, and folic acid have little effect). Requirements also emerge for glycine (spared by serine), valine and isoleucine (their ratio is critical; leucine and threonine assist in maintaining a good balance), and, at very slightly higher temperatures, phenylalanine, tryptophan, cystine, and lysine. Requirements for Mg, Fe, Zn, and Mn appear to rise steeply with temperature; metal toxicities have to be circumvented carefully. The proportion of histidine + arginine to carbohydrate has to be increased, and a Krebs-cycle component such as succinic acid becomes stimulatory. At 36.3–36.7°, a further supplement of crude natural materials such as an autoclaved suspension of Ochromonas cells is needed. Relevance of these findings to fever stress in vertebrates, general mitochondrial function, and repair of radiation damage, is discussed.

Notes: SUMMARY. According to Kahl the trichostome genus Trichopelma may be considered to include five species: T. sphagnetanim, T. eurystoma, T. euglenivora, T. opaca, and T. torpens. A sixth species is added in the present account. The following constant characteristics distinguish it from the five earlier described ones: total lack of trichocysts; differences in the morphology of the body surfaces; dissimilarity of the upper and lower surfaces in shape (one plane, the other convex), in ciliation, and in number of cuticular furrows; unique location of the contractile vacuole.

Notes: SUMMARY. Isospora citelli n. sp. is described from the rock squirrel, Citellus variegatus Utah, from Grand Canyon National Park, Arizona. Its oocysts are subspherical, 22.4 by 21.5 μ, with a smooth, two-layered wall, an oocyst refractile globule and a sporocyst residuum, but without a micropyle or oocyst residuum.

Notes: SUMMARY. Leptomonads of Leishmania tarentolae were grown continuously in a defined medium containing: inorganic salts', glucose, hemin, 17 amino acids, purines and pyrimidines, and a mixture of vitamins of the B group. In this medium the population of organisms reached about 20 to 50 million per ml. alter 1 week at 27°C. Only slightly better growth occurred in a partially defined medium containing bovine plasma fraction V. In earlier experiments, however, omission of the plasma fraction resulted in decreased growth, and under these circumstances cholesterol or lecithin had growth-stimulating effects. In later experiments in the fully-defined medium no effect of these lipids could be found. The leptomonads were shown to require at least the following substances: inorganic salts; a source of purines and pyrimidines; tryptophan and the nine other amino acids essential for the growth of rats, glutamic acid, tyrosine, proline, serine, one or more of the group alanine, glycine and aspartic acid; folic acid, biotin, pantothenic acid, nicotinamide, riboflavm, thiamine, and either pyridoxine plus choline or pyridoxal or pyridoxamine. Choline at 2 × 10−5 m gave optimal growth in the presence of pyridoxine at 1 × 10−5 m. In a medium with a suboptimal concentration of choline (0.4 × 10−5 m) the leptomonads grew through nine transfers but they were mostly somewhat rounded and aflagellate.

Notes: SUMMARY. Trichopus lachmanni n. sp. possesses, in common with other members of the family Dysteriidae, a fixation organelle composed of vesicles and a secretory ampoule. But no “foot-like” appendix exists in this species, and the ampoule opens at the bottom of an antapical pit which bears a short fringe of membranelles derived from the somatic ciliature. The vibratile fringe participates in the spinning of the glutinous secretion which temporarily fastens the organism to the substrate. This specialized ciliature is characteristic of the genus Trichopus which was created by Claparède & Lachmann for a species, T. dysteria, which, though insufficiently described, is certainly different from T. lachmanni.

Notes: SUMMARY. Various species of ciliates are characterized by the formation and accumulation in the cytoplasm of mineral concretions which are refringent, isotropic or anisotropic. These cytoplasmic inclusions most often are composed of calcium carbonate; in several species, however, their nature remains partially or even totally undetermined. The isotropic calcium-containing concretions often exhibit a definite shape; the calcium carbonate in this case appears to be bound to an organic substrate. The physiological role of the calcic concretions is not known; their characteristic presence in a given species is not necessarily related to ecological conditions. In a few species the calcification is localized in definite structures: spicules, skeletal plates, or otoliths of organelles supposedly sensory in nature.

Notes: SUMMARY. The combination Tween 80 and cholesterol replaced blood serum as a requirement for the cultivation of Tritrichomonas foetus. Choline + potassium glycerophosphate enhanced growth in the presence of Tween 80 + cholesterol. A method for ensuring adequate cholesterol suspension-an essential factor for consistent growth-is described.

Notes: SUMMARY. High variability within cultures of Tritrichomonas batrachorum (from Bufo boreas halophilus, Rana catesbeiana, and Rana pipiens) casts doubt on the validity of length measurements for species designation.The morphology differs from that described previously. The axostylar capitulum is a complex structure with a preblepharoplastic portion and a perinuclear cup spreading laterally in curved membranes ending in filaments. The parabasal apparatus is Y-shaped with a very short base.Two types of amoeboid activity occur. Filopodia are used for anchorage. Their bases migrate and the filopodia may fuse and reseparate as in foraminifers. Ingestion is an amoeboid process utilizing lobopodia. Food adherent to a lobopod is invested in a food cup at the surface. Ingrowth of the lip of the food cup forms the food vacuole, or in cases of cannibalism, constricts the prey and forms a food tube pushing a vacuole before it. Repeated, this produces an alternating series of vacuoles and tubes. There is a pseudo-amoeboid degenerative process in which the flagella, withdrawn into the cytosome, continue to move and produce non-functional lobopod-like protrusions.An actual cytostome has not been observed, but there is a region of reduced staining intensity ventral to the axostylar capitulum.

Notes: . Norlevinea n. g. is established for microsporidia in which a uninucleate meront changes into a sporont by secreting a thin, membranous, sporontogcnetic and fragile sporophorous vesicle (pansporoblast membrane) in which four uninucleate sporoblasts are formed. In contrast to the genus Gurleya, the sporoblasts and later the spores are permanently joined into doublets, being laterally cemented by an electron-dense substance structurally identical to and continuous with the exospore layer. The polar filament is of the anisofilar type. The type species is Norlevinea daphniae (Weiser, 1947) n. comb., a parasite of the ovaries of Daphnia longispina occurring in several carp ponds in Czechoslovakia.

Notes: . The microsporidian parasite known as Nosema helminthorum Moniez, 1887, parasitic in the tapeworm Moniezia expansa (Rudolphi, 1810), has been shown by electron microscopy to have two cycles of development, one with isolated nuclei, the other with paired nuclei (diplokarya). Both merogony and sporogony of the two separate sequences take place in direct contact with the host cell cytoplasm and ultimately give rise to unikaryotic and diplokaryotic sporoblasts. Sporogony is disporoblastic. The nuclear condition of the spores was not seen. The sequences, corresponding to those of the genera Unikaryon and Nosema, may be part of a single dimorphic life cycle and, if so, the species will have to be transferred to a new genus.

Notes: . Sarcocystis falcatula Stiles, 1893 is re-described. Intermediate hosts of the parasite which was earlier described as Sarcocystis debonei Vogelsang, 1929 are species of passeriform, psittaciform, and columbiform birds. In these birds, muscle zoites are 6.88 × 2.19 (4.8-8.4 × 1.2-3.6) μm and are enclosed in a cyst wall with regular protrusions, 1-5 μm long. The convoluted primary wall has multiple thin areas in the osmiophilic layer. Microtubules originate in the ground substance and extend to the tips of the protrusions. The only known definitive host is the opossum, Didelphis virginiana; rats, cats, a dog, and a ferret could not be infected from muscle cysts. Sporocysts from opossums infected from five different infected avian sources measure 11.2 × 7.4 (9.6−12.0 × 6.0-8.4)μm.

Notes: . Descriptions are given of two new species of Hepatozoon Miller, 1908 found in the pygmy squirrel, Idiurus macrotis, in the Ivory Coast. Gamonts of both are parasites of monocytes.The size and shape of the gamonts of one, H. normani n. sp., are similar to those of a number of gamonts of other species of rodent hemogregarines and the separate identity of the parasite is based on the host restriction of mammalian hemogregarines. The gamonts of the other species, H. dolichomorphon n. sp., are remarkably long and slender and are unlike those of any other known hemogregarine of mammals. Schizonts of this species were found in a smear prepared from heart blood.

Notes: . The presence of nonvariant antigens (NVAs) limited to bloodstream forms of Trypanosoma brucei brucei and Trypanosoma brucei rhodesiense was demonstrated for the first time by immunodiffusion and Immunoelectrophoresis. Noncloned and cloned populations were employed in preparation of polyclonal antisera in rabbits and of antigens to be used in the immunologic reactions. The NVAs could be shown best in systems in which hyperimmune rabbit sera (adsorbed with procyclic forms to eliminate antibodies against antigens common to bloodstream form and procyclic stages) were reacted with trypanosomes characterized by heterologous variant-specific antigens (VSAs).The NVAs demonstrated in this study are very likely different from the common parts of VSAs. As has been suggested by experiments with living trypanosomes, at least a part of the NVAs appears to be located on the surface of the bloodstream forms. In these experiments involving the quantitative indirect fluorescent antibody test, the amount of fluorescence recorded for the heterologous system, i.e. ETat 5 trypanosomes incubated with anti-AmTat 1.1 serum, equalled ∼3.0% of the fluorescence emitted by the AmTat 1.1 bloodstream forms treated with their homologous antiserum. Evidently, only small amounts of NVAs are present on the surfaces of T. brucei bloodstream forms.In addition to the NVAs, the electrophoresis results suggested the presence of antigenic differences between procyclic stages belonging to different T. brucei stocks.

Notes: . Late schizonts from continuous cultures of P. falciparum were concentrated over Percoll, inoculated to various experimental media at the rate of about 20 × 106 per 0.5 ml of medium, and incubated in a candle jar at 37° for 1 day. Controls in standard culture medium showed a heavy invasion with young rings in the previously uninfected red cells introduced with the inoculum of schizonts. In a medium of high potassium content containing a 33% extract of human erythrocytes, this invasion was inhibited and many free merozoites were present. If, however, this same medium was supplemented with both ATP, as the dipotassium salt at 1.6 mM, and sodium pyruvate at 3.6 mM, there appeared large numbers of extracellular forms resembling young rings. Examination of these by electron microscopy shows that they are indeed merozoites that have begun to differentiate extracellularly. This suggests that the trigger for differentiation of merozoites may not depend on the process of entry into a red cell but rather on specific factors within the red cell.

Notes: . Opossums (Didelphis marsupialis), act as intermediate hosts for Besnoitia darlingi and could be infected orally with sporozoites (oocysts) and bradyzoites (tissue cysts), or intraperitoneally (i.p.) with tachyzoites. Infections could presumably be transmitted through cannibalism. Cats (Felis catus), the definitive host, could be infected only with bradyzoites but not sporozoites. Oocysts shed by cats measure about 12 × 12 μm, resemble similarly sized oocysts of Toxoplasma gondii and Hammondia hammondi, and must be differentiated by the appearance of tissue cysts after experimental infection of intermediate hosts. Cats did not form tissue cysts of B. darlingi. Tachyzoites from the related B. jellisoni could be used in the Sabin-Feldman dye test to determine the development of antibody to B. darlingi in opossums after infection.

Notes: Developmental stages of Caryospora simplex were found in connective tissue of the cheek, tongue, and nose of Swiss-Webster and C57 BL/6 mice (Mus musculus) from 8 through 70 days after oral inoculation with 50,000 or 250,000 oocysts, or 60,000 free sporocysts of the same species obtained from an Ottoman viper, Vipera xanthina xanthina. The earliest developmental stages were seen on day 8 post-inoculation (PI) and consisted of two types of meronts and gamonts (undifferentiated sexual stages). Gamonts, microgametocytes, macrogametes, and unsporulated oocysts were found on days 10 and 12 PI. Fully sporulated, thin-walled oocysts containing eight sporozoites surrounded by a thin sporocyst membrane were first seen 12 days PI. Monozoic cysts (caryocysts) were first seen 12 days PI and appeared fully viable throughout the duration of the study, 70 days PI. Four mice injected intra-peritoneally with 150,000 free sporozoites and killed 12 days PI contained unsporulated and sporulated oocysts in connective tissues of the cheek, tongue, and nose, suggesting that sporozoites may be carried to the site of infection via the lymphatic/circulatory system. Four cotton rats, Sigmodon hispidus, inoculated orally with 250,000 oocysts all had unsporulated and sporulated oocysts of C. simplex in connective tissue of the cheek, tongue, and nose when killed on day 12 PI, indicating extraintestinal development in the secondary host is not species specific. This is the first report of a heteroxenous coccidium with both asexual and sexual development in the primary (predator) and secondary (prey) hosts.

Notes: Trypanosoma lucknowi n. sp. was isolated in culture from one of 126 Macaca mulatta originating from the vicinity of Lucknow, Uttar Pradesh, India. Trypanosoma lucknowi is distinctive because of the large number of epimastigotes and trypomastigotes which, in culture, exhibit no movement or only a slight bending of the flagellar end. This limited motility coincides with a free flagellum which is either completely absent or rudimentary. The microorganism is cloned readily, and the description is based upon such cultures. Trypanosoma lucknowi shows pronounced differences from other trypanosomes of South Asian macaques and from “aflagellar” African trypanosomes. The ultrastructural demonstration of a cytostome and contractile vacuole suggests ultimate grouping with stercorarian trypanosomes. A 3-D reconstruction of the flagellar pocket/cytostome region is included.

Notes: The ultrastructure of the freshwater, heterotrophic dinoflagellate Peridiniopsis berolinense (Lemm.) Bourrelly resembles other dinoflagellates in the structure of its nucleus, theca, flagella, and mitochondria. Other features less frequently reported in related organisms include fine sub-sulcal fibers, collared pits in the flagellar base region, and unusual structures herein termed fibrillar lamellae. Numerous vesicles are present, some of whose contents are distinctly crystalline, while others contain what appears to be membranous material arranged in either whorls or parallel stacks; still other vesicles contain electron-dense, granular spheres. Of particular interest is the transitional helix present in the longitudinal flagellum, this being the first report of such a structure among the dinoflagellates. Plastids of any kind are lacking, and a peduncle is present and is used during phagotrophy.

Notes: Several axenic strains of pathogenic and nonpathogenic Entamoeba histolytica were tested for their capacity to digest native radioactive type I collagen gels and to produce liver abscesses when injected into the liver of newborn hamsters. The results demonstrate that the pathogenic strains of amebas (HM1:IMSS, HM3:IMSS, HM38:IMSS, and HK9) have a collagenolytic activity that closely correlates with their in vivo capacity to produce liver lesions. The nonpathogenic isolate (Laredo) did not show collagenolytic activity and failed to produce lesions in the liver of newborn hamsters. The results also demonstrate that type I collagen obtained from rodents and cats is degraded less by amebic collagenase than is bovine collagen, which is similar to human collagen. These findings suggest that species susceptibility to invasive infection may depend, among other factors, on the characteristics of the extracellular components of host tissues.

Notes: At Makthlawaiya, in the Paraguayan Chaco, the prevalence of Trypanosoma (Schizotrypanum) cruzi infection among both domestic Triatoma infestans and domestic dogs was 38%, and IgG anti-T. cruzi antibody was detected by the quantitative enzyme-linked immunosorbent assay (ELISA) in 80% (105/133) of human sera. Ninety percent (25/28) of T. cruzi strains isolated from both T. infestans and dogs showed heterozygous isoenzyme profiles for glucose phosphate isomerase, phosphoglucomutase and 6-phosphogluconate dehydrogenase. These strains appeared to be closely related to Bolivian zymodeme 2. Three Paraguayan T. cruzi strains showed homozygous isoenzyme profiles, similar to those of major Brazilian zymodemes. It was concluded that T. cruzi strains with heterozygous isoenzyme profiles predominate in domestic transmission cycles in this highly endemic area of the Paraguayan Chaco.

Notes: Results obtained in immunofluorescence localization studies involving three antisera, six species of ciliates, and a variety of fixation procedures suggest that superior results can often be obtained by fixing cells in 35–70% ethanol. Formaldehyde fixation appeared to induce redistributions of epiplasmic proteins and surface antigens which were not observed in ethanol-fixed cells. In addition, background fluorescence was significantly lower in ethanol-fixed cells than it was in cells fixed in aldehydes.

Notes: Actin has been identified in the ciliated protozoon Tetrahymena paravorax on the basis of the ultrastructural detection of filaments typically decorated with heavy meromyosin (HMM) in glycerinated microstome cells. These filaments are widely distributed in endoplasmic and cortical regions and can form bundles. They are particularly numerous in elongating cells; HMM-binding filaments run approximately parallel to rib microtubules in the ectoplasm of the right wall of the buccal cavity and seem to extend to the cytopharyngeal region, suggesting some role of actin in maintenance of the crest-trough pattern of ribbed wall and/or in formation of food vacuoles. Extensive actin bundles are observed below some membranellar areas and are thought to follow the course of the microtubular “deep fiber bundle.” The “fine filamentous reticulum” underlying the oral ribs and the “apical ring” extending beneath kinetosomes of ciliary couplets display filaments that do not bind HMM and are ˜ 14 nm in diameter. No evidence for actin in these structures was obtained in the present study. The “specialized cytoplasm” of the cytostome-cytopharyngeal region appears as an undecorated reticulum with 20 nm-spaced nodes. Occasionally HMM-binding filaments were found inside the macronucleus, just beneath its envelope. Actin is suggested to be involved in cell shaping and in control of the transport of food vacuoles.

Notes: Two allelic Mendelian mutations which confer a short flagella phenotype were used to explore flagellar size control in Chlamydomonas reinhardtii. When mutant/wild type quadriflagellate dikaryon cells were constructed, their two short flagella rapidly grew out to near wild type length. The kinetics of elongation suggest that the flagellar assembly process is not intrinsically self-limiting as a number of otherwise attractive models for size control require. Instead, we suggest that there exists a cellular machinery dedicated to flagellar size control and that the short-flagella mutations alter the machinery in some as yet unknown way. One of the mutants shows temperature-sensitive flagellar assembly, and both are flagellaless in acetate media. Genetic analysis indicates that the temperaturesensitive, acetate-sensitive, and short-flagella phenotypes have a common genetic basis. The responsible gene has been named shf-1, and it has been mapped to chromosome VI, approximately 5 map units from the centromere.

Notes: Fourteen strains of Naegleria australiensis, including the type strain, were compared for virulence for mice, maximum growth temperature, lectin agglutination, isoenzyme pattern, and total protein banding pattern. Their relation to other species of Naegleria also was compared by immunoelectrophoretic analysis. Strains with high virulence, comparable to that of N. fowleri, were found to be different in concanavalin A agglutination as well as with regard to zymograms and total protein patterns. Although serologically different from N. fowleri and reacting with N. australiensis antiserum in the fluorescent antibody test, these high-virulence strains differed in number of immunoelectrophoretic precipitin bands. Because of these results, the high-virulence strains are considered to be a subspecies of N. australiensis. The low-virulence strains showed minor differences from the type strain. Thus, N. australiensis does not appear to be as homogenous a species as N. fowleri. Pathogenic N. australiensis also seems to be more widespread than previously thought.

Notes: In Pleurotricha lanceolata, the ventral somatic infraciliature presents 13 frontoventral cirri, 5 transverse cirri, one row with 18–19 left marginal cirri and two rows of right marginal cirri of different length. On the dorsal side there are six longitudinal rows of dorsal bristles, four of them bipolar and the other two less than half body length. The oral infraciliature includes the adoral zone of membranelles, with 45–55 membranelles of three or four rows of kinetosomes each, and two undulating membranes (paroral and endoral membranes), each with two rows of kinetosomes. Some structures of the oral and somatic fibrillar systems have also been examined and are similar to those described in other species of hypotrichous ciliates.

Notes: Fecal samples of 36 ground squirrels, Spermophilus beldingi, from Tioga Pass (elev. ca. 3315 m) in the Sierra Nevada, California, yielded oocysts of Eimeria beckeri in nine squirrels, E. citelli in four squirrels, E. beldingii n. sp. in two squirrels, and degenerated, unidentifiable oocysts in ten squirrels. Eimeria beldingii n. sp. oocysts are ellipsoidal, 30–34 × 24–30 (mean 32 × 26) μm with a two-layered, rough, striated wall, without a micropyle or residuum, with polar granules; they contain ellipsoidal or ovoid sporocysts 11–15 × 9–12 (mean 13 × 10) μm with a Stieda body and residuum.

Notes: Ten years of research on digestive vacuoles (phagosomes) of Paramecium caudatum have revealed sequential changes both within the vacuole lumen as well as within the surrounding membrane. Four vacuole stages can be recognized by a combination of thin section and freeze-fracture ultrastructural features. Three sets of vesicles (discoidal vesicles, acidosomes, and lysosomes) fuse with the vacuole, each at a predetermined stage, to bring about these membrane and physiological changes. At various times membrane is removed as vesicles from the vacuole surface, which has the effect of regulating vacuole size. Membrane recycling, membrane replacement, and specific membrane to membrane recognition all appear to be operating during the digestive cycle. Details of these events are summarized in this address and a number of unanswered questions suggest areas for future research.

Notes: Reports of Cryptosporidium in various hosts and cross-transmission experiments are reviewed. Cryptosporidium has been found in mammals (Primates, Artiodactyla, Perissodactyla, Carnivora, Lagomorpha, and Rodentia), birds, reptiles, and fish. The only cross-transmission attempts that have been made have been from mammals to other mammals and to a few birds. Names have been given to 19 “species,” but it is concluded that only four of these should be considered valid at present. These are: C. muris Tyzzer, 1907 in mammals, C. meleagridis Slavin, 1955 in birds, C. crotali Triffit, 1925 in reptiles, and C. nasorum Hoover, Hoerr, Carlton, Hinsman & Ferguson, 1981 in fish.

Notes: In vitro excystation of sporozoites of the heteroxenous coccidian Caryospora simplex Léger, 1904 (Apicomplexa: Eimeriorina) is described. Sporocysts freed mechanically from oocysts released a maximum of 51% of their sporozoites within 45 min at 25°C and a maximum of 74% within 20 min at 37°C when incubated in a 0.25% (w/v) trypsin–0.75% (w/v) sodium taurocholate (bile salt) excystation solution. At emergence from sporocysts, sporozoites were weakly motile then became highly active after about 2 min in excystation solution. Sporozoites within sporocysts exposed to bile salt only became highly motile within 25 min at 25°C and within 15 min at 37°C but did not excyst. When exposed only to trypsin at the above temperatures, the Stieda body dissolved; the substieda body remained intact, and the sporozoites exhibited only limited motility within sporocysts; only a few excysted. Intact, sporulated oocysts incubated at 25° or 37°C in 0.02 M cysteine-HC1 and a 50% CO2 atmosphere for 18 h had no morphologic changes in the oocyst wall. Further incubation of these intact oocysts in excystation solution for 30 min at 37°C caused neither motility of sporozoites within sporocysts nor excystation. Grinding oocysts for 30 sec in a motor-driven, teflon-coated tissue grinder caused motility of some sporozoites within sporocysts but did not result in excystation.

Notes: The effect of the cationic permeant fluorescent dye, rhodamine 123 (R123), on the in vivo growth of Plasmodium yoelii was examined. Plasmodium yoelii-infected mouse erythrocytes were incubated in vitro with R123 and injected intravenously into mice. Examination of daily parasitemias showed that R123 delayed parasite growth whereas rhodamine 110, a neutral compound, and fluorescein, a negatively charged fluorescent dye, did not. Infected erythrocytes treated with R123 were not cleared from the circulation even 7 h after injection. Quantitation of cell-associated R123 by spectrophotometry revealed that infected cells with increased levels of R123 considerably prolonged the 2% prepatent period, the time required for the parasite to develop a 2% parasitemia. Degenerating parasites within and outside the host erythrocytes were observed on day 1 of infection in the mice. Thus it follows that R123, which accumulated in infected erythrocytes, inhibits the growth of P. yoelii; moreover, when R123-labeled infected erythrocytes were treated with 1–10 μM carbonylcyanide m-chlorophenylhydrazone (CCCP), a proton ionophore, to release R123 from the cells, the inhibitory effect on the growth rate of P. yoelii was partially reversed.

Notes: Oocysts of Caryospora corallae n. sp. were isolated from the feces of three Emerald Tree Boas Corallus caninus. The spherical oocysts of C. corallae averaged 22.4 μn (range 18.7 to 24.6) in diameter and were lacking a micropyle and oocyst residuum; a polar granule was present. The ovoid sporocysts measured 19.1(17.6-20.0) × 13.1(11.7-14.0) μm and a sporocyst residuum and a Stieda body were present. The oocyst wall was approximately 1 μm thick. The sporulation was completed in about 5–6 days at 23 ± 2°C. This is the first report of the genus Caryospora from Corallus caninus a member of the Boidae.

Notes: One third of a collection of cloned Stylonychia pustulata micronuclear DNA PstI fragments were found to be of a similar size, consistent with their being members of a repetitious sequence family with a repeat size of about 160 base pairs. Cross-hybridization experiments confirmed that these small cloned fragments are related by sequence homology. Hybridization of the cloned repetitious sequences to PstI digested micronuclear DNA revealed a “ladder” of bands (step size = 160 base pairs), indicating that the repeats are found in tandem arrays. This is the first demonstration of highly repetitious, tandemly repeated sequences in a ciliated protozoan.

Notes: The erythrocytic developmental cycle of Plasmodium falciparum can be conveniently divided into the ring, trophozoite, and schizont stages based on morphology and metabolism. Using highly synchronous cultures of P. falciparum, considerable variation was demonstrated among these stages in sensitivity to chloroquine. The effects of timed, sequential exposure to several clinically relevant concentrations of chloroquine were monitored by three techniques: morphological analysis, changes in the rate of glucose consumption, and changes in the incorporation of 3H-hypoxanthine into parasite nucleic acids. All three techniques gave essentially identical results. The trophozoite and schizont stages were considerably more sensitive to the drug than ring-stage parasites. Chloroquine sensitivity decreased as nuclear division neared completion. The increase in chloroquine sensitivity was coincident with a marked rise in the rate of glucose consumption and nucleic acid synthesis. The rate of nucleic acid synthesis decreased as schizogony progressed while glucose consumption continued at high rates during this process. The degree of chloroquine sensitivity was not highly correlated with either metabolic activity.

Notes: Bloodstream trypomastigote and culture procyclic (insect midgut) forms of a cloned T. rhodesiense variant (WRATat 1) were tested for agglutination with the lectins concanavalin A (Con A), phytohemagglutinin P (PP), soybean agglutinin (SBA), fucose binding protein (FBP), wheat germ agglutinin (WGA), and castor bean lectin (RCA). Fluorescence-microscopic localization of lectin binding to both formalin-fixed trypomastigotes and red cells was determined with fluorescein isothiocyanate (FITC)-conjugated Con A, SBA, FBP, WGA, RCA, PNA (peanut agglutinin), DBA (Dolichos bifloris), and UEA (Ulex europaeus) lectins. Electron microscopic localization of lectin binding sites on bloodstream trypomastigotes was accomplished by the Con A-horseradish peroxidase-diaminobenzidine (HRP-DAB) technique, and by a Con A-biotin/avidin-ferritin method. Trypomastigotes, isolated by centrifugation or filtration through DEAE-cellulose or thawed after cryopreservation, were agglutinated by the lectins Con A and PP with agglutination strength scored as Con A 〈 PP. No agglutination was observed in control preparations or with the lectins WGA, FBA or SBA. Red cells were agglutinated by all the lectins tested. Formalin-fixed bloodstream trypomastigotes bound FITC-Con A and FITC-RCA but not FITC-WGA, -SBA, -PNA, -UEA or -DBA lectins. All FITC-labeled lectins bound to red cells. Con A receptors, visualized by Con A-HRP-DAB and Con A-biotin/avidin-ferritin techniques, were distributed uniformly on T. rhodesiense bloodstream forms. No lectin receptors were visualized on control preparations. Culture procyclics lacked a cell surface coat and were agglutinated by Con A and WGA but not RCA, SBA, PP and FBP. Procyclics were not agglutinated by lectins in the presence of competing sugar at 0.25 M. The expression of lectin binding cell surface saccharides of T. rhodesiense WRATat 1 is related to the parasite stage. Sugars resembling α-D-mannose are on the surface of bloodstream trypomastigotes and culture procyclics; n-acetyl-D-galactosamine and D-galactose residues are on bloodstream forms; and n-acetyl-D-glucosamine-like sugars are on procyclic stages.

Notes: The cationic permeant fluorescent dye rhodamine 123 (R123) was used to stain Plasmodium yoelii-infected mouse erythrocytes. Fluorescence microscopic observations demonstrated that the parasite, but not the matrix of the infected erythrocyte, accumulated the dye. Differences in fluorescence intensity could not be found at the various developmental stages of the parasite; however, quantitation of the cell-associated dye revealed an increase in R123 uptake with parasite development. The retention of the parasite-associated dye, as measured by fluorescence microscopy and spectrophotometry after extraction of R123 with butanol, was markedly reduced by treatment of the infected erythrocytes with a proton ionophore, carbonyl cyanide m-chlorophenylhydrazone (CCCP), and an inhibitor of proton ATPase, dicyclohexylcarbodiimide (DCCD). These results indicate that the accumulation and retention of R123 in P. yoelii reflect the parasite membrane potential and suggest that the parasite plasma membrane has a membrane potential-generating proton pump.

Notes: Distinctive organic-walled resting cysts of at least three different types with a highly conservative morphology appear to characterize specific orders or groups of genera within the Class Polyhymenophorea (Protozoa, Ciliophora), contrasting markedly with the great diversity of form seen in trophic stages. Polyhymenophorean ciliates have been considered in the past to form a cohesive class within the Phylum Ciliophora and, possibly, to represent the pinnacle of ciliate evolution. Evidence from cysts challenges the cohesive nature of the class, suggesting that the hypotrichs should be subdivided and that they have a different phylogenetic origin from the heterotrichs, tintinnids, and oligotrichs.

Notes: Yellow-brown, algal symbionts varying in diameter from approximately 5 μ m to 20 μ m, associated with solitary Radiolaria with spongiose skeletons (i.e. Spongodrymus sp.), exhibit fine structural features resembling the Prymnesiida (botanical class, Prymnesiophyceae). A large central vacuole is surrounded by a thin layer of cytoplasm containing plastids with lamellae composed of three thylakoids and granular pyrenoids with internal tubules immersed between the thylakoids. The pyrenoids lack internal thylakoid membranes. The nucleus is surrounded by a dilated cisterna of the nuclear envelope that also encloses the plastids and gives rise to saccules of the endoplasmic reticulum. The algal symbionts appear coccoid; hence no flagella nor surface scales were observed. The symbiont fine structure is compared to similar yellow-brown symbionts associated with Acantharia. Thus far, three kinds of algal symbionts have been observed to be associated with solitary Radiolaria: dinoflagellate, prasinomonad, and this apparent prymnesiomonad.

Notes: Ultrastructural observations of the cortically-located mitochondria of Tetrahymena thermophila revealed associations not only between the mitochondria and certain of the cortical microtubule bands, but also between the mitochondria and the epiplasm of the cortex. Most of the distal mitochondrial surface is close and parallel to the epiplasm; favorable views show bridge-like structures spanning the 20–10 nm gap between the mitochondrion and the epiplasm.Previous studies have shown that the placement of mitochondria in the cortex appears to be determined by certain of the cortical microtubule bands. This study, however, shows that mitochondrion-microtubule interactions account for only a small proportion of the total mitochondrial area associated with the cortex; the rest is accounted for by the epiplasm. A possible analogue of the spectrin layer of erythrocyte membranes, the epiplasm may be important in helping to arrange the intricately organized components of the ciliate cortex. Its involvement in apparently helping to “moor” mitochondria to their cortical sites is the first suggestion of any role in cell patterning played by the epiplasm.

Notes: Ce Tetradimorpha, rencontre en eau douce se présente soit sous forme sphérique pourvue de quatre flagelles et d'axopodes rayonnants, soit sous forme allongée avec a l'avant quatre flagelles associes a quatre axopodes et a l'arriére six a huit axopodes divergents. L'etude ultrastructurale révèle un cytosquelette axopodial de type centroplastidie comprenant un centroplaste lenticulaire homogéne, centre organisateur des quatre axopodes anterieurs et des six a huit axopodes posterieurs, auquel s'ajoutent les quatre cinetosomes des flagelles anterieurs. En outre, un deuxiéme éleément cytosquelettique incluant un microtubule associe chacun des quatre cinetosomes a l'axopode antérieur correspondant. Des cordons microfibrillaires réunissent axopodes et cinetosomes au niveau du centroplaste, puis a quelque distance du centroplaste les axopodes posterieurs. Les axonémes des axopodes comprenant de 5 a 30 microtubules sont constitues de triades, lorsqu'on peut détecter une organisation. Le noyau, a nucléole central est coince dans le cone axopodial posterieur, lui-méme entouré des dictyosomes. Par l'organisation du cytosquelette, par la structure des kinétocystes, par la structure des flagelles dépourvus de mastigonémes tubulaires, Tetradimorpha différe nettement de Ciliophrys marina. Comme le prévoyait Davidson (1975), il represönte bien un des chainons dans la série évolutive des Héliozoaires centrohélidiens. Mais il ne présente guère d'affinites avec les Chrysomonadines considerees comme la souche des Héliozoaires. L'intéret de ce Protiste dans l'étude de la differentiation et de l'evolution du cytosquelette est également présente.〈section xml:id="abs1-2"〉〈title type="main"〉ABSTRACTThis freshwater species of Tetradimorpha has a spherical body with four flagella and radiating axopods; it transforms into a pear-shaped cell that anteriorly has four flagella intercalated between four axopods and posteriorly has six to eight divergent axopods. Ultrastructural study reveals an axopodial cytoskeleton of the centrohelidan type comprising an homogeneous lenticular centroplast which acts as MTOC for axopodial microtubules. A second skeletal element is a microtubular linkage between the kinetosomes and the axonemes of anterior axopods. A microtubule embedded in dense material diverges from near the base of each kinetosomes and parallels the distal portion of the axoneme of each anterior axopod. A microfibrillar envelope around the centroplast links the axopodial bases to the kinetosomes situated just above. Close to the centroplast, microfibrillar strands link the axopodial axonemes to the kinetosomes. Axopodial axonemes are composed of 5 to 30 microtubules irregularly arranged except for some that form equilateral triangles. The nucleus containing a central nucleolus is constrained within a cone formed by the axonemes of the posterior axopods and surrounded by dictyosomes. By the cytoskeletal organization, the structure of kinetocysts, and flagella wthout tubular mastigonemes, Tetradimorpha differs obviously from Ciliophrys marina. As Davidson (1975) predicted, Tetradimorpha is an intermediate link in the centrohelidan lineage: however, it lacks the characteristics of chrysomonads, the supposed ancestors of Heliozoa. The contribution of this genus to the study of the differentiation and the evolution of the cytoskeleton is also presented and discussed.

Notes: Two-dimensional gel electrophoresis was used to identify the patterns of protein synthesis during initiation, and the patterns of membrane protein expression following initiation, in all of the mating types of the Tetrahymena thermophila B family. In addition, one-dimensional analysis was used to survey 125I-Concanavalin A-binding proteins. Although a large number of proteins was identified by each technique, no variation among the mating types was observed.

Notes: Cells of Tetrahymena pyriformis, T. thermophila, and Euglena gracilis were saturated with nitrogen gas at pressures up to 300 atm and rapidly decompressed. Damage was assessed by measuring post-decompression cell fragmentation or viability. Occurrence of intracellular bubbles was determined by cinephotomicrography performed during the decompression or by direct observations afterwards. The extreme gas supersaturations induced led to intracellular bubble formation and rupture in cells of Tetrahymena that contained food vacuoles, but only with supersaturations of 175 atm or higher; 225 atm left few cells intact. Bubbles were never observed in cells of Euglena or in Tetrahymena cells freed of food vacuoles, even when they were decompressed from substantially higher nitrogen supersaturations. Cells of Euglena were most resistant and were unaffected by supersaturations up to 250 atm.

Notes: The rapid, synchronous differentiation of N. gruberi from amoebae to flagellates is a useful paradigm to study aspects of cell differentiation, including regulation of the synthesis of proteins that are related to the changes in cell shape and motility, which occur during differentiation. The differentiation requires synthesis of new RNA and protein molecules to accomplish defined morphogenetic events. Specific new proteins, including the tubulins that form the flagellar microtubules, are synthesized at various times during differentiation, and particular mRNA species appear and disappear. The time course of the synthesis of the α and β subunits of flagellar tubulin is paralleled by the programmed appearance and disappearance of flagellar tubulin mRNAs. The evidence supports the hypothesis that the synthesis of flagellar tubulin is regulated by the transcription, and subsequent disappearance, of flagellar tubulin mRNA. Translatable mRNAs for two calmodulin-like calcium-binding proteins appear and disappear contemporaneously with those for flagellar tubulin. During differentiation the synthesis of actin, the major protein of amoebae, is selectively shut down, and translatable actin mRNA rapidly disappears. This description of the orderly appearance, utilization, and disappearance of the mRNAs for actin, calcium-binding proteins, and flagellar tubulin during differentiation provides means and motivation to investigate the mechanisms that regulate these events.

Notes: Earlier experimental work involving macronuclear implants in Stentor coeruleus has shown that the cytoplasmic cortex of the nuclear site 1) attracts the macronucleus and 2) holds it in place during interphase. Now experiments indicate macronuclei transferred with overlying cortex elongate in the direction of the transferred cortical pigment stripes, whether or not the transferred stripes realign in the direction of the host stentor's stripes. Therefore the third function of the cortex is to determine the direction of elongation and thus assure that both daughter cells at division receive part of the macronucleus.

Notes: During an electron microscopic study of Glugea stephani, three morphologically distinct tubular appendages that are continuous with the sporoblast plasmalemma were observed. The tubules were designated as: type I, 45–50 nm in diameter and 600–900 nm in length; type II, 25–35 nm in diameter, averaging 1300 nm in length; type III, 50–70 nm in diameter and with an indeterminate length, which often exceeds 3000 nm. Type III tubules contain regularly spaced, electron-dense particles that are approximately 30 nm in diameter. Since many genera of microsporida have some type of appendage, which may eventually be utilized for taxonomic purposes, we propose the formation of a system of serially numbered detailed descriptions of these structures to promote uniformity and clarity in future publications.

Notes: Cells of Paramecium tetraurelia, stock hrd, cultured in a micro-capillary containing 1 μl fresh culture medium, expressed mating activity through the whole cell cycle. Mating-reactive G2 phase cells can conjugate with cells of other phases. The G2 phase cells, which have double (4C) the normal micronuclear DNA content, undergo pre-meiotic DNA synthesis when conjugated with G1 phase cells. The micronucleus of the progeny from the cross between a G1 and a G2 cell becomes triploid.

Notes: Buffered solutions of KCl and NaCl were tested for their stimulatory effect on the germination of variously-aged spores of Vavraia culicis. Germination was optimal in 0.2 M KCl, pH 6.5 for one isolate, and, for another isolate, peaks of germination occurred at pH 7.0 and 9.5. Spores incubated for several hours in suboptimal solutions became unable to germinate under optimal conditions. After being returned to water, they regained their ability to germinate. Calcium chloride, magnesium chloride, and ammonium chloride inhibited germination. After ingestion by mosquito larvae, spores germinated near the posterior end of the midgut.

Notes: . Leishmania tropica promastigotes transport α-aminoisobutyric acid (AIB), the nonmetabolizable analog of neutral amino acids, against a substantial concentration gradient. AIB is not incorporated into cellular material but accumulates within the cells in an unaltered form. Intracellular AIB exchanges with external AIB. Various energy inhibitors (amytal, HOQNO, KCN, DNP, CCCP, and arsenate) and sulfhydryl reagents (NEM, pCMB, and iodoacetate) severely inhibit uptake. The uptake system is saturable with reference to AIB-and the Lineweaver-Burk plots show biphasic kinetics suggesting the involvement of two transport systems. AIB shares a common transport system with alanine, cysteine, glycine, methionine, serine, and proline. Uptake is regulated by feedback inhibition and transinhibition.

Notes: . Oxytricha strains used in biochemical studies have traditionally been grown in unaerated, unagitated culture tubes or Fernbach flasks. These cultures are limited in volume to about one liter and have a very nonuniform distribution of cells, with the majority of the cells at the very top or bottom of the medium. We have found conditions in which Oxytricha can be grown in 50-liter fermentation vats. The cultures grow to a uniform density of about 6000 cells/ml.

Notes: . The fine structure of the tomite of Foettingeria actiniarum (Claparède) was examined and compared with that of other apostome tomites. This stage in the life cycle has a unique configuration of kineties that form a spiral through the cytoplasm in the interior of the body. The structure and behavior of this internal spiral were evaluated as a mechanism for the storage of kinetosomes, an adaptation to the ciliate's two-host life cycle. The spiral is composed of nine ribbons of laterally compressed kinetosomes that are in contact with a thin electron-dense fibril. Paralleling the kineties of the spiral are conspicuous, swollen lamellae of the rough endoplasmic reticulum; these lamellae contain moderately electron-dense material. The spiral is associated with the large contractile vacuole and winds about the macronucleus. The tomite of Foettingeria possesses a single, robust, caudal cilium located in a pit, along with the nozzle-like pore of the contractile vacuole. The walls of the pit contain several trichocysts arranged radially about the caudal cilium and aimed into the pit.

Notes: . Fine structural studies of a specialized vesicle system associated with the endoplasmic reticulum (ER) of exo-erythrocytic Plasmodium berghei suggest that this system may be the equivalent of a Golgi apparatus. Patches of ER, randomly distributed in the cytoplasm of developing parasites, are formed of smooth and ribosome-studded cisternae intermingled with each other. The vesicle systems are located between as well as at the edges of ER aggregates and appear to be in different stages of budding from the cisternae. Prolonged osmication reveals distinct staining of the nuclear envelope and ER of the parasites as well as part of the Golgi apparatus of the hepatocytes. However, the small vesicles associated with the parasite's ER are unstained, as are the coated vesicles in the Golgi region of the liver cell. These sites in the parasite cytoplasm seem comparable to the concave surface of the Golgi apparatus in liver cells. The pinched-off vesicles fuse with others to form the prominent peripheral vacuolization characteristic of the nearly mature exo-erythrocytic form. The formation of these peripheral vacuoles and their subsequent fusion with the parasite membrane may be an exocytosis mechanism supplying the rapidly expanding parasite with new plasma membrane material.

Notes: . Ultrastructural cytochemical techniques were used to analyze the nucleus and the kinetoplast of epimastigotes of Trypanosoma cruzi. With the use of ethanolic phosphotungstic acid, which detects basic proteins, reaction product was seen in the chromatin and at the periphery of the kinetoplast. Thallium alcoholate, which interacts with DNA, stained strongly the whole kinetoplast and the chromatin. With the use of a silver impregnation method that detects acidic nucleolar proteins, silver granules were seen preferentially located in the central region of the nucleolus. With the EDTA method, which reveals the presence of ribonucleoproteins, staining was observed in the nuclear pores. Also 6–8 nm fibrils, 25 nm and 40 nm granules, which correspond to the perichromatin fibers, interchromatin granules and the perichromatin granules, respectively, were identified in the nucleus. The EDTA method also revealed the presence of 40 nm granules in the kinetoplast. These granules were seen mainly at the two extremities of the kinetoplast. Freeze-fracture images indicate that the nuclear membrane contains ca. 9 pores/μm2 of nuclear surface area. The mean diameter of the pores was 80 nm. All these results suggest that epimastigotes of T. cruzi have a very active nucleus and a high rate of nucleocytoplasmic interchange.

Notes: SUMMARY. A study has been made of the cytology of an undescribed species of Spirostomum. The species is much smaller than S. ambiguum. The peristome extends to about half the length of the animal, whereas in S. ambiguum it is about two-thirds the body length. The nuclear apparatus reveals some striking differences. The macronucleus is cylindrical, and not chain-like, in the vegetative animal. The micronuclei are far fewer than in S. ambiguum and number 6–15, but are larger in size. During binary fission, the macronucleus becomes condensed into an oval or polymorphic mass and is drawn out again into a cylinder before it is cut into two lengths. The micro-nuclei divide by mitosis and, whatever their number in the vegetative animal, only 7–8 take part in the division. The others presumably degenerate.

Notes: SUMMARY. The structure and life history of a new species of eugregarine, Gregarina garnhami n.sp., is described from the intestinal caeca and mid-gut of Schistocerca gregaria Forsk. The parasite destroys considerable areas of the caecal epithelium and in cases of heavy infection, the masses of parasites present in the mid-gut result in the formation of localized barriers between the gut wall and the food material in the lumen. The cephalont and sporont stages of gregarines from Locusta migratoria migratorioides R. & F. and Anacrydium aegyptium Linn. are shown to be similar to those from Schistocerca gregaria and are believed to belong to the same species.

Notes: SUMMARY. Oxygen concentrations of 70–80 per cent of an atmosphere destroy all chromosomes of the flagellate Trichonympha provided the oxygen treatment is carried out during the early stages of gametogenesis at which time the chromosomes are in the process of duplicating themselves. This treatment does no damage to the cytoplasm and its organelles. Following the loss of chromosomes, the centrioles function in the production of the achromatic figure, the flagella, and‘parabasal bodies. Then the cytoplasm divides, thus producing two anucleate gametes which make some progress in the cytoplasmic differentiations characteristic of normal male and female gametes of Trichonympha.It is also possible, with somewhat higher concentrations of oxygen, with temperatures slightly above the freezing point and a longer period of treatment, to destroy the chromosomes of resting asexual nuclei in several genera of the flagellates that live in the roach Cryptocercus. So far as one can determine by observing organisms so treated, their cytoplasm and organelles are not injured.