ALBERT

Notizen: SYNOPSIS. In an electron microscopic study, counts of peripheral microtubules were made in spheromastigotes and trypomastigotes in tissue cultures of embryonic heart muscle cells. In interphase spheromastigotes there were, at the level of the nucleus, ∼ 114 microtubules; in dividing forms, there were ∼ 222. In trypomastigotes, the number of microtubules varied according to the level of the section—there were fewer than 40 tubules in the pointed ends of an organism, while in the central segment the number of these elements ranged from 60 to 115. The highest number of microtubules was found in the region containing the Golgi complex. The distance between the microtubules was constant, equalling 44 nm, even at the pointed ends of a trypanosome. This suggests that the microtubules course parallel to one another. Cross sections and randomly arranged, variable length, longitudinal sections of the tubules were noted around the kinetosomes in dividing organisms.

Notizen: SYNOPSIS. An intracellular protozoon was discovered in the epithelium of young rainbow trout (Salmo gairdneri) exposed for as short a time as 1 hr to water known to contain infective stages of Myxosoma cerebralis. Light- and electron-microscopic examination of this tissue revealed what appeared to be a proliferative stage (presumptive schizont) of a sporozoon; other possible stages in the life cycle were also observed. The relationship of this unidentified protozoon to M. cerebralis remains unresolved.

Notizen: SYNOPSIS. Coriphosphine staining of Stylonychia mytilus exconjugants at different times after separation reveals some details of the developing macronucleus. Green fluorescence is seen in both bands and heterochromatic blocks of the polytene chromosomes. No red fluorescence was observed along these chromosomes. Fragments of the old macronucleus and the pycnotic micronuclei have red or orange fluorescence. Red fluorescence is characteristic also of nucleoli in the new macronucleus.

Notizen: SYNOPSIS. Trophozoites and other pre-spore stages of the myxosporidan Myxosoma cerebralis were taken from infected rainbow trout (Salmo gairdneri) and cultured in vitro. Cultures eventually yielded mature spores capable of discharging their polar capsules. This is the first report of culture of a myxosporidan.

Notizen: SYNOPSIS. Conjugation in Paramecium tetraurelia can be induced within mating-reactive cultures of a single mating type by treating the cells with solutions of KC1 + acriflavine in culture medium low in Ca2+. Gene mutations with known physiologic effect were used as selective inhibitors of cell surface membrane function to see which functions are necessary for chemical induction of conjugation. The results strongly suggest that a transient increase in the internal concentration of calcium at the very beginning of chemical induction is a necessary but not sufficient step.

Notizen: SYNOPSIS. In a method for isolating a relatively clean suspension of concentrated Plasmodium berghei ookinetes from infected midguts of Anopheles stephensi at appropriate times after the infective blood meal, the ookinetes are freed from the midguts by enzymatic digestion, and then concentrated by means of a BSA/renografin gradient. The mean number of ookinetes recovered/midgut was 152. More than 95% of the recovered ookinetes were viable by the criteria of motility, incorporation of adenosine and leucine, and appearance on light and electron microscopic examination. Trypan blue exclusion was not a valid criterion for viability. These ookinetes were not useful for in vitro studies of further development due to the presence of contaminating microorganisms. Our attempts to determine their potential for further development in vivo have similarly not been successful. Nevertheless, our ability to obtain large numbers of ookinetes at defined times during development now permits further studies on their structure, biochemistry, and physiology, as well as comparison with ookinetes formed in vitro.

Notizen: SYNOPSIS. An extracellular surface coat was observed at the fine-structural level on the outer lamina of the pellicular and flagellar membranes of intact Trypanosoma musculi bloodstream forms. The surface coat had a mean width of 9.2 nm, and was composed of a somewhat electron dense, uneven, fibrillar-like matrix. Brief trypsin treatment of living blood forms completely removed the cell surface coat.Several cytochemical methods applicable to electron microscopy were used to detect the presence and distribution of carbohydrates in the trypanosome's surface coat and pellicular membrane. The polycationic dye compounds employed were: ruthenium red, ruthenium violet, Alcian blue chloride, and lanthanum nitrate. Electron-dense stain reaction products, indicative of polysaccharides, were evident in the surface coat of cells treated with these dyes, which also agglutinated both living and glutaraldehyde fixed cells. Like the surface coat, the pellicular membrane of trypsinized cells gave strong positive staining reactions with the several dyes, indicating the presence of membrane bounded carbohydrates, and living and glutaraldehyde-fixed trypsinized cells were agglutinated with the polycationic stains. Bloodstream forms were treated with the enzymes, α-amylase, dextranase, and neuraminidase. No obvious morphologic difference, however, was apparent between the surface coat of untreated cells and those subjected to treatment with any of the various glycoside hydrolase enzymes. Further, these enzymes had no apparent gross effect on the staining affinity of the surface coat for the several polycationic dyes. Cationized ferritin was used to visualize the negative cell surface charge of T. musculi bloodstream forms. Large quantities of cationized ferritin were bound in the surface coat matrix. Glycoside hydrolase enzyme treatments had no apparent effect on the amount of ferritin bound in the surface coat. Cationized ferritin was bound also to the outer lamina of the pellicular membrane in trypsinized cells, which had quantitatively less ferritin bound per surface unit area than bloodstream forms untreated by the enzyme. Living and glutaraldehyde-fixed cells were agglutinated with cationized ferritin. The results obtained in the various experiments indicated that polyanionic polysaccharides were constituent terminal ligands of the surface coat matrix and pellicular membrane in T. musculi bloodstream forms.

Notizen: SYNOPSIS. The sporogonic stages of Leucocytozoon dubreuili in the midgut and salivary glands of the simuliid vectors was studied by electron microscopy. Young uninucleate oocysts have a pellicle that initially resembles that of the ookinetc. Numerous electron-dense bodies and microtubules in the peripheral cytoplasm may be involved in the formation of the cyst wall. The dense bodies appear to give rise to the amorphous material of the wall. The tubules which run circumferentially beneath the oocyst's boundary probably serve as a skeletal support for the cell surface during deposition of the wall material. A subcapsular “space” which provides area for expansion of the developing sporozoites is formed in early multinucleate oocysts. The subcapsular “space” appears to be formed through a condensation of the peripheral cytoplasm, resulting in an osmotic gradient across the oocyst's limiting membrane. Consequently water diffuses out, creating a fluid-filled space. Sporozoite formation begins with localized thickenings on the oocyst's limiting membrane. Subsequent extension of the thickened regions into the subcapsular “space” marks the onset of sporozoite budding. The process is highly synchronized, and culminates with the production of up to 150 sporozoites about the sporoblastoid body. The structure of sporozoites from mature oocysts and of the salivary glands of the vector is basically similar, although salivary gland sporozoites are more elongate and have numerous electron-dense micronemes. The paired rhoptries in the latter sporozoites are more elongate and uniformly electron-dense than in oocyst sporozoites.

Notizen: SYNOPSIS. Temperature strongly influenced morphogenesis of intracellular trypomastigotes in cell culture infected with 2 different strains of T. cruzi. With the Gilmar strain the amastigote-to-trypomastigote differentiation readily occurred at 33 and 37 C, whereas with the CL strain differentiation took place at 33 C but was inhibited at 37 C. The possibility of this selective thermosensitivity resulting from mutational adaptation of the parasite is discussed.

Notizen: La présence de diplocaryons à tous les stades de la sporulation d'Octosporea muscaedomesticae Flu, 1911 est mise en évidence ainsi qu'une membrane pansporoblastique autour des sporoblastes en développement et des spores. Le problème présenté par ces 2 faits est discuté. Une évolution étrange des ribosomes périnucléaires et démontrée. Une telle étude ultrastructurale est nécessaire et intéressante car O. muscaedomesticae est l'espèce-type du genre Octosporea.

Notizen: SYNOPSIS. It is evident from studies of cellular recognition resulting in phagocytosis by Chaos carolinensis that cilia from certain protozoa serving as prey, e.g. Tetrahymena pyriformis, influence phagocytic action. Glutaraldehyde-fixed C. carolinensis to which cilia, isolated from Tetrahymena, are attached by the benzidine reaction, are engulfed by living Chaos but not digested. Fixed Chaos without attached cilia are not engulfed. Adding increased numbers of cilia from Tetrahymena progressively inhibits the rate at which Chaos phagocytizes Paramecium caudatum, suggesting blockage by the cilia of contact sites normally available to initiate phagocytosis of Paramecium.

Notizen: SYNOPSIS. The microsporidan Glugea stephani is a common parasite of juvenile English sole (Parophrys vetulus) in Yaquina Bay, Oregon. Field observations indicated that fish became infected only in the upper estuary where summer temperatures were above 15 C and the incidence of infection reached 79.8% in the late fall.Laboratory infections developed and parasite growth occurred only at or above 15 C. The parasite was successfully transmitted to juvenile English sole by brine shrimp (Artemia salina) and amphipod (Corophium spinicorne) vectors as well as by direct ingestion of spores by the host. Infections that resulted from ingestion of spore-carrying vectors were much heavier than those resulting from the direct ingestion of spores. The speckled sanddab (Citharichthys stigmaeus), a nonpleuronectid flatfish, and chum salmon (Oncorhynchus keta) were refractory to G. stephani infection in the laboratory.Results of this study suggest that G. stephani is potentially lethal to young pleuronectid flatfishes when heavy infections involve the entire intestine and reduce the capacity to absorb nutrients. Under these circumstances, starvation is probably the direct or indirect cause of death. The restriction of infection to fish that reach the upper estuary very likely mitigates the impact of G. stephani caused mortality on the entire English sole population on the Yaquina Bay nursery ground.

Notizen: SYNOPSIS. Optimal assay conditions are described for 8 hydrolases of Euglena gracilis var. bacillaris, SM-L1 (streptomycinbleached) strain, 7 of which have an acid pH-optimum. Acid phosphatase, β-galactosidase, β-glucosidase, β-fucosidase, cathepsin D, RNase, DNase, and an esterase are active in cell homogenates. Amylase has very low activity, and β-glucuronidase, arylsulfatase, β, N-acetyl-glucosaminidase, α-fucosidase, and α- and β-mannosidase are inactive.Hydrolase activity increases as a culture proceeds from the midexponential to the late stationary-phase of growth, being most pronounced in the case of β-glucosidase. In cultures deprived of a utilizable carbon source, the specific activities of the hydrolases (per mg total protein or dry weight) increase. When expressed on a per cell basis, however, the activities of DNase decrease while those of β-galactosidase, cathepsin D, and RNase increase. The hydrolases appear to be involved in the adaptation of Euglena to the metabolic demands imposed by different conditions of growth.

Notizen: SYNOPSIS. Sensitivity to pentamidine of bloodstream forms and culture forms of Trypanosoma brucei brucei, strains of this subspecies, and strains of T. brucei rhodesiense characteristically differs in vitro. Analyses of transport parameters for pentamidine uptake in these organisms show differences that correspond with drug sensitivity. Long slender bloodstream forms of T. b. brucei have a high affinity for the drug and high rates of uptake as indicated by Km and Vmax values for [3H]pentamidine transport. Although pentamidine and stilbamidine resistance is associated with dyskinetoplasty. this condition does not itself confer resistance to pentamidine nor does it affect pentamidine transport. However, drug-resistant strains show lower rates for pentamidine transport as does T. b. rhodesiense, which is characteristically less sensitive to the drug. Of all the forms and strains studied, procyclic trypomastigotes were least sensitive to pentamidine and had a remarkable ability to exclude the drug.

Notizen: SYNOPSIS. The 16S ribosomal RNA of the chloroplast of Euglena gracilis strain Z has been characterized in terms of its 2-dimensional electrophoretic “fingerprint” (T1 ribonuclease). Over 100 spots were resolved on the “fingerprint” and each spot was characterized as to which RNA oligonucleotide fragment(s) it contained. When compared to similar analyses of prokaryotic 16S rRNAs and eukaryotic cytoplasmic 18S rRNAs, the chloroplast 16S rRNA was a typically prokaryotic RNA, but bore little if any relationship to eukaryotic 18S rRNAs. Therefore, the cistrons for chloroplast 16S rRNA are related to the equivalent prokaryotic cistrons, but, apparently, are not related to the equivalent eukaryotic cistrons. Among the organisms available for comparison, the Euglena chloroplast 16S rRNA appears most closely related to the 16S rRNA of the eukaryote, Porphyridium cruentum (a red alga), and at least distantly related to the 16S rRNAs of the blue-green algae and perhaps also to the bacilli.

Notizen: SYNOPSIS. A survey of 22 Western Painted Turtles (WPT), Chrysemys picta belli Gray; 2 Map Turtles (MT), Graptemys geographica Le Sueur; 8 False Map Turtles (FMT), Graptemys pseudogeographica Gray; 26 Illinois Mud Turtles (IMT), Kinosternon flavescens spooneri Smith; and 5 Spiny Softshell Turtles (SST), Trionyx spiniferus Le Sueur, revealed the presence of 10 species of coccidia. The species found, the shape and average length-width dimensions (in micrometers) of their oocysts, and the respective hosts from which they were isolated were as follows: Eimeria chrysemydis Deeds & Jahn, slightly pear-shaped (27.6 × 17.0), from 5 WPTs; Eimeria delagei marginata Deeds & Jahn, markedly pearshaped (22.1 × 17.6), from 2 WPTs, 1 MT, and 3 FMTs; Eimeria graptemydos sp. n., broadly ellipsoid to subspherical (12.6 × 11.4), from 10 WPTs, 2 MTs, and 5 FMTs; Eimeria lutotestudinis sp. n., broadly ellipsoid to subspherical (11.9 × 10.8), from 4 IMTs; Eimeria mascoutini sp. n., broadly ellipsoid to subspherical with outer surface of oocyst wall mammillated (14.0 × 11.9), from 2 SSTs; Eimeria mitraria (Laveran & Mesnil), asymmetrically ellipsoid, bearing 1 or 2 conical projections at the polar end and 2 or 3 at the antipolar end (10.0 × 7.6), from 7 WPTs, 1 MT, 3 FMTs, and 4 IMTs; Eimeria pseudogeographica sp. n., narrowly ellipsoid to slightly ovoid (19.5 × 13.5), from 5 WPTs and 3 FMTs; Eimeria tetradacrutata sp. n., spherical to subspherical with outer surface of oocyst wall mammillated (19.5 × 19.2), from 1 WPT; an Eimeria sp. narrowly ellipsoid, bearing 2 conical projections at the polar end and 2 at the antipolar end (23.8 × 13.2), from 1 MT; and Mantonella hammondi sp. n., conical to narrowly ellipsoid, bearing 1 conical projection at the polar end and 2 or 3 at the antipolar end (14.3 × 7.1), from 1 IMT. The overall infection rate for the 63 turtles examined was 54.0%.

Notizen: SYNOPSIS. The schizont stage of 3 strains of the testate marine ameba, Trichosphaerium sp., was examined in the transmission electron microscope. The cytoplasm of this multinucleate organism contained the usual organelles; the test was covered by spicules. Pseudopodial types included broad ectoplasmic lobopods that assisted in locomotion, and thin dactylopods that probably had a sensory function. Nuclear division, observed in one of the strains, was characterized by an intact nuclear envelope (at least through anaphase), and the absence of centrioles. Nuclei in an ameba divided synchronously. An unusual intranuclear body of unknown function was found in another of the strains examined.

Notizen: SYNOPSIS. The fine-structural aspects of development of microgamonts of Eimeria magna were studied in kidney cell cultures and in experimentally infected rabbits. Spheroidal masses of gamont-like cytoplasm containing ribosomes, polyribosomes, and amylopectin granules were found within the parasitophorous vacuole; these bodies were apparently pinched off the surface of the gamont. Nucleoli were present in the early stages of nuclear division but disappeared as development proceeded. Spindles were eccentric and the nuclear membrane always remained intact in dividing nuclei. Nuclei eventually became elongate in shape, compact, and electron dense at the end oriented toward the periphery and lucent centrally. Usually, only the dense portion was incorporated into the gamete as the gamete developed by protruding from the gamont surface. Fullyformed microgametes were biflagellate and contained a nucleus, a mitochondrion, and 8-10 microtubules. Multiple-membrane complexes, which apparently originated from the nuclear envelope or endoplasmic reticulum, were found within the gamont residual body. Occasionally, 2 or 3 micro- and/or macrogamonts were seen within the same cell. In some cells one or 2 micro- or macrogamonts as well as several merozoites were present in the same parasitophorous vacuole.

Notizen: SYNOPSIS. Oxytricha fallax and Stylonychia pustulata possess 6 rows of dorsal bristle units. Each dorsal bristle unit consists of a pair of kinetosomes; the anterior kinetosome has a cilium and the posterior kinetosome a ciliary stub. The kinetosome pair, located at the bottom of a cortical pit surrounding the cilium and ciliary stub, is surrounded by an asymmetrical fibrillar mass. Future rows 1-4 are formed from 2 sets of primordia originating within mature dorsal rows 1-3. Rows 5 and 6 originate from the anterior regions of both right marginal cirral primordia. Old dorsal bristle units utilized in formation of primordia are presumably maintained in the new rows of the proter and opisthe; those outside the primordia are resorbed. The morphogenetic pattern of the Oxytrichidae is similar to those of the Urostylidae and Holostichidae, but quite different from that of the Euplotidae.

Notizen: . Allantosoma intestinalis, a suctorian ciliate isolated from the intestine of the horse, was studied utilizing light and electron optical methods. These small sausage-shaped organisms have a varying number of tentacles (between one and 12) located at each extremity of the body. The microtubular axoneme of each tentacle in cross-section consists of two files of microtubules arranged in a daisy-like configuration. Haptocysts occur in the tentacle shaft, abutted to the plasma membrane of the knob of the tentacle, and in the cell body. The haptocysts are bottle-shaped, with prominent annular striations around their midportion. The cell is covered by three membranes, an outer plasma membrane, an outer alveolar, and an inner alveolar membrane. A thin epiplasmic layer is found beneath the inner alveolar membrane, and a single row of microtubules underlies the epiplasm. The subpellicular microtubules are arranged parallel to each other forming a corset around the cell along the long axis: such a system is not characteristic of suctorians. A field of diminutive kinetosomes (each 180 nm long, max. of 15 per field), lacking cilia, was found below the cortex. The function of these prokinetosomes is unknown. A ciliated swarmer has not been observed, only the nonciliated adult. The characteristics of Allantosoma are compared with those of other suctorian genera.

Notizen: . Temperature shifts have been used to block critical points in the conjugation sequence of Paramecium tetraurelia. Increasing temperatures above 27°C reduced ciliary agglutination, pair formation, and nuclear exchange; a complete inhibition of these stages occurred at 37°C. Temperatures below 19°C had no effect on ciliary agglutination or nuclear exchange but completely inhibited pair formation. The bases for the cells’ inability to form pairs at 19°C and 37°C were sought. Cells placed below 19°C were unable to deciliate or fuse membranes in the holdfast region; at 37°C, membrane fusion in both the holdfast and paroral regions was prevented. Time course studies on cross-fertilization reveal that temperatures 35°C block all stages of the process up to the actual exchange of pronuclei. After the exchange has begun, the process continues despite the elevated temperature. Temperature shifts are discussed as a means of conditionally blocking critical points in the developmental program of conjugation.

Notizen: . [14C]chimyl and [3H]batyl alcohols were added to Crithidia fasciculata cultures during the mid-log phase of cell growth, and the lipid extracts of the cells were analyzed for degradation products. C. fasciculata cells were able to take up exogenous glyceryl ethers, and in amounts as high as the endogenous lipid content. The glyceryl ether taken up by the cells was incorporated into lipids either prior to the ether bond cleavage or after degradation to fatty acid. The extent of degradation and the degree of incorporation of degradation products into cellular lipid were higher for chimyl than for batyl alcohol. Batyl alcohol was not metabolized efficiently, leading to the formation of large intracellular pools of free substrate. One product of glyceryl ether degradation was identified as alkyl-dihydroxy acetone, and was detected inside and outside of the cells. The data strongly suggest that this product is the first stable intermediate in the degradation process and indicate that the extracellular formation of alkyl-dihydroxy acetone is due to the action of exocnzyme; ecteted by the cells. The constant detection of alk I cnyl glycerol among the degradation products indicates the existence of a second mechantsm in C. fasciculata for converting the alkyl-to alkenyl-glycerol.

Notizen: . Sick budgerigars from a local aviary were found to be infected with Giardia. The trophozoites were of the Giardia duodenalis type as defined by Filice with elongate median bodies pointed on one or both ends and more or less perpendicular to the long axis of the body. Using three fixation-staining methods, and material from three birds, the length ranged from 10 to 18 μm and the width from 4.5 to 11 μm with a mean length to width ratio of 2. Attempts to culture the trophozoites in vitro from intestinal scrapings were unsuccessful. Also attempts to transmit the infection by fecal cysts to canaries and mice failed. It is proposed that the budgerigar form be called Giardia duodenalis, race psittaci.

Notizen: Three species of Pseudocohnilembus were examined with respect to infraciliature and the silverline system. The freshwater species P. putrinus and P. pusillus exhibited no differences in general organization of the argyrophilic structures when compared with the marine P. marinus. A review of the Pseudocohnilembus species that have been described on the basis of silver preparations shows that at present four species have been well characterized: P. pusillus (Quennerstedt, 1869) nov. comb., P. putrinus (Kahl, 1928) nov. comb., P. hargisi Evans & Thompson, 1964, and P. marinus Thompson, 1966, P. persalinus Evans & Thompson, 1964 and P. longiseta Evans & Thompson, 1964 are regarded as identical with the older known P. pusillus, because with respect to morphology and argyrophilic structures they are within the range of variability of that species; and their names thus fall as synonyms of P. pusillus.

Notizen: The structure of micronemata arising from the surface of the bloodstream form of Trypanosoma gambiense was studied by electron microscopy. In order to produce micronemata, trypanosomes were incubated in either 1) phosphate buffered saline supplemented with glucose (PBSG), 2) immune mouse serum or 3) PBSG after passage through a DEAE-cellulose column. Electron microscopic examination of the parasite revealed the presence of thread-like micronemata arising from the anterior end and from the flagellar pocket regardless of the incubation conditions. Negative staining revealed a distinct peripheral fringe layer with nodular protrusions covering the entire surface of the micronema. The distribution and number of intramembrane particles (IMP) on the P and E faces of the micronema were similar to those of the flagellum of T. gambiense, indicating a close relationship between the membrane structure of the micronema and the flagellum. Micronemata became fragmented and adhered to each other after incubation of the parasite in the media for 12 h. Since micronemata tend to have the characteristics of adhesiveness and fragmentation, fragments of these structures might adhere to various host organs. Dispersal of potential antigenic material might be responsible, in part, for the induction of the host immune response.

Notizen: The motion of Paramecium caudatum has been investigated at various temperatures by measuring the transient behavior of spatial distribution in the diffusion process of organisms that, by electric stimulus, are initially gathered at a single place in the glass culture cell. The spatial distribution through the course of diffusion has a nearly Gaussian profile. Dispersion was obtained at 1 sec intervals and increased linearly with time. The time dependence of the dispersion gave a diffusion coefficient for the random motion of the organisms. The results show that the diffusion coefficient has a maximum at the temperature at which the paramecia were cultivated.

Notizen: . Conjugating cells of Euplotes vannus (syngens Naples and Barcarès) were investigated for Con A-binding sites by means of fluorescence microscopy. Cells fixed with 0.4% paraformaldehyde and incubated with 15 μg/ml FITC-Con A showed a distinct region of strong fluorescence, the size, shape and localization of which characteristically changed during the course of conjugation, first appearing at courtship stage (cells contact without forming pairs) and vanishing soon after the migration pronuclei have been exchanged, but before cells have separated. Con A binding and conjugation were blocked by cyclohexamide and α-galactosidase. Con A-binding was also inhibited by α-mannosidase, α-glucosidase and β-galactosidase without affecting conjugation except a delay of pair formation. The results suggest an involvement of newly formed or translocated glycoconjugates in cell pairing during conjugation.

Notizen: . The development of Sarcocystis cruzi was studied in an 11-day-old calf killed seven days postinoculation with 5 × 108 sporocysts from feces of coyotes. Uninucleate zoites were found in arteries of mesenteric lymph nodes but not in other organs. Zoites measured 4.9 × 3.0 (3.5–7.0 × 2.1–3.5) μm. Of the 36 zoites studied, 31 were in endothelial cells, four were in macrophages in the lumen of arteries, and one was free in the lumen of an artery. Infected endothelial cells were two to three times larger than uninfected cells. Zoites appeared structurally similar to sporozoites. The occurrence of zoites in macrophages suggests that sporozoites of Sarcocystis might use such cells to reach the site of their first merogony.

Notizen: . The enzyme that catalyzes the critical conversion of the proposed reductive thymidine catabolic end product, P-aminoisobutyric acid, to the initial anabolic reutilization substrate, probably methylmalonic semialdehyde, was investigated to un- ambiguously substantiate the operation of a reductive pyrimidine catabolic and reutilization pathway in Tetrahymena pyriformis strain GL. Although most of the studies on the further metabolism of P-aminoisobutyric acid in other organisms suggested transamination of p-aminoisobutyric acid to methylmalonic semialdehyde followed by its further oxidation to methylmalonic acid, such a transaminating system could not be demonstrated. By means of a sensitive fluorometric assay system, however, a low, but significant amount of P-aminoisobutyric acid oxidase activity was detected. This enzymatic activity exhibited the following characteristics in homogenates: good activity in alkaline 0.2 M Tris-HCI buffer with a rather broad pH optimum ranging from 7.8 to 9.0; optimum activity at a temperature of 37°C; stimulation on the addition of exogenous FAD; inhibition on the addition of divalent cations, EDTA, or PCMB; little stimulation on the addition of detergents; and no increase in activity on repeated freezing and thawing. In addition, crude preparations of this oxidase were found to be relatively stable when stored up to one week either refrigerated or frozen, to have a specific activity of 2.8 nmoles/min/mg of protein, and to have a K, of 3.6 × 10− M for d,l-β-aminoisobutyric acid. Whether this high Km, is physiologically significant or not, however, will have to await further investigation. Preliminary (NH4)2SO4 fractionation and affinity chromatography studies also indicated that this enzyme appears to be a unique and specific oxidase whose activity is separable from other marker enzymes, including other oxidases.

Notizen: . Phosphatase activity in Trypanosoma rhodesiense has been examined histochemically by light and electron microscopy and by enzymatic assay in homogenate fractions. Using a method with lead as capture ion, acid phosphatase was found in lysosome-like vesicles and in the flagellar pocket. No alkaline adenosine triphosphatase (ATPase) was detectable by this method. Direct assay of p-nitrophenylphosphatase activity in homogenate fractions showed that acid phosphatase activity was strongly membrane-bound, but that activity at pH 9 was minimal in both soluble and particulate fractions. “Endogenous” ATPase activity was localized specifically and reproducibly in the mitochondrial membranes and under the plasma membrane of the flagellum. This nonenzymic reaction product could not be eradicated by glycerol extraction or glucose depletion. Unlike the membrane staining, which was manifest only after lead treatment, heat-resistant electron-dense material was found in the matrix of lysosomal vesicles in trypanosomes fixed in glutaraldehyde only and not subjected to further treatment with heavy metal reagents. X-ray emission analysis showed the presence of calcium and phosphorus, indicating that the matrix might have a phosphate storage function.

Notizen: The longitudinal flagellum of Ceratium tripos moves in two dissimilar ways: undulation and retraction. The undulatory wave is planar and has a wavelength of 74.3 ± 9.6 μm and an amplitude of 14.2 ± 2.3 μm in sea water. The beat frequency is 30 Hz at 20°C, pH 8.0. The retractile motion is unique to Ceratium and is triggered by mechanical stimulation on the cell body, especially at the tip of the apical horn. When it retracts, the longitudinal flagellum folds every 4–5 μm along the flagellum. Cinematographic study showed that the flagellum folded from tip to base and was finally installed into the sulcus, a groove on the ventral side of the cell. This motion is completed in sea water within 28 msec. The retracted flagellum then re-extends and restores the undulation within a few seconds. The flagellum unfolds in the proximal portion first, then the distal, and finally the middle portion. Fixation always triggers the retraction. Scanning electron microscopy showed that the flagellum is folded and secondarily twisted in a helix. A new fiber in addition to the flagellar axoneme was found in the retracted flagellum by phase microscopy. This fiber (R-fiber) seems to contract during the retraction to fold the flagellum.

Notizen: Phagocytosis of culture forms of Trypanosoma cruzi was assayed by a radioisotopic method. Purified polymorphonuclear leukocytes (PMN) were mixed with 3H-uridine-labeled T. cruzi epimastigotes in the presence or absence of anti-T. cruzi antibodies. The reaction was stopped by adding N-ethyl-maleimide, and noningested parasites were lysed by complement. The percentage of radioactivity incorporated into the PMN pellet was recorded. The phagocytosis reaction was rapid, yielding maximum incorporation at 30 min at which point the radioactivity associated with the PMN cells decreased through release of the isotope to the supernatant. The degree of incorporation of radio-labeled parasites was a function of the effector/target cell ratio and the antibody concentration. The method is suitable for the quantitative determination of phagocytosis of T. cruzi by normal PMN.

Notizen: The following species of the gregarine genus Lankesteria were found in a study of ascidians from Monterey Bay, on the central California coast of the Pacific Ocean: L. abbotti n. sp. in Clavelina huntsmani, L. aplidii n. sp. in Aplidium solidum, L. ascidiae in Ciona intestinalis, L. diaphanis n. sp. in Archidistoma diaphanes, L. euherdmaniae n. sp. in Euherdmania claviformis, L. montereyensis n. sp. in Archidistoma molle, L. pescaderoensis n. sp. in Ritterella rubra, L. pittendrighi n. sp. in Ascidia ceratodes, L. psammii n. sp. in Archidistoma psammion, L. ritterellae n. sp. in Ritterella pulchra, L. ritterii n. sp. in Archidistoma ritteri, L. synoici n. sp. in Synoicum parfustis, and Lankesteria sp. in Botrylloides sp. No gregarines were seen in the following ascidians: Aplidium californicum, A. propinquum, Botryllus tuberatus, Diplosoma macdonaldi, Perophora annectens, and Polyclinum planum.

Notizen: The effect of holothurin (a marine biotoxin) on the resistance of mice to Trypanosoma musculi was measured by studying changes in the parasite population in vivo. Swiss Webster (SW), Beige (BG), and Black (BL) mice treated with holothurin prior to and simultaneously with infection of trypanosomes had lower parasitemias than controls. Higher levels of parasitemia were observed in mice treated after infection with trypanosomes. The timing of administration of holothurin appeared to be an important factor in the observed effect. The minor variations in the parasitemia seemed to be related to the mouse strain.

Notizen: The structure of the cyst wall of the heliozoon Echinosphaerium nucleofilum has been investigated using light microscopy, scanning and transmission electron microscopy, and X-ray microanalysis. The cyst wall is a composite structure of seven or eight layers. These are: an enveloping gelatinous layer; a layer of siliceous spheroidal bodies; an electron-dense supporting membrane; a broad electron-lucent zone; an electron-dense layer; a layer of helicoidally packed material; and one or two layers with a granular appearance lying next to the plasma membrane of the encysted organism. The structure of the cyst wall closely resembles that of Actinophrys sol, confirming the close relationship of these actinophryid heliozoa while emphasizing their distinctiveness from other amoeboid protista.

Notizen: Morphogenesis of cell division was investigated in Diophrys scutum, D. oligothrix, and D. appendiculata utilizing both light microscopy of living and stained specimens and SEM of preserved specimens. The cortical morphogenetic pattern of Diophrys is similar to that of other members of the family Euplotidae. The opisthe oral primordium, which develops in a subsurface pouch, forms posterior to the parental buccal cavity. The proter inherits the parental adoral zone of membranelles (AZM) apparently unchanged. The endoral membrane forms to the right of the posterior end of the AZM in the proter, in association with the developing AZM in the opisthe. The paroral cirrus and membrane develop from a single streak that first appears along the right edge of the buccal cavity in the proter to the right of the developing buccal structures of the opisthe. Frontal and transverse cirri develop in both proter and opisthe from five separate cirral primordia that form to the right of the buccal cavity. Left marginal cirri do not develop in association with the corresponding parental structures. Kinetosomes formed within the opisthe oral primordium, or kinetosomes that were part of any parental ciliary structure, do not appear to become part of any developing paroral structures, frontal, transverse, or left marginal cirri. Speciation within the genus Diophrys and evolution of the family Euplotidae as they relate to the morphogenesis of cortical structure are discussed.

Notizen: Autecology (cellular organelles and secretions, encystment and dispersal abilities), environmental conditions (physiological tolerances and interaction with other organisms), and evolutionary history contribute to protist biogeography, which manifests ecological and historical aspects. Ecological biogeography is seen in the influence of geochemistry on the distribution of fresh-water phytoflagellates and algae, and of moisture and vegetation type on soil-litter protists. A temporal feature is often present because many protists encyst and respond only to certain ranges of temperature and organic content. Historical biogeography has occurred by radiative host evolution on symbiotic protozoa (e.g., termite flagellates and rumen ciliates), and by the isolating effects of water currents, temperature, and density gradients on oceanic protists (coccoliths, dinoflagellates, foraminifera, radiolaria, and tintinnines). These two aspects combine in protists living on animal surfaces. Humans affect protist biogeography by increasing parasite ranges through human migrations and by water pollution. They can diminish these situations by disease control and exploiting appropriate ciliates in sewage disposal. Many free-living protozoa appear to be cosmopolitan, but mating types and isoenzyme studies suggest that speciation with its geographic connotations may be more widespread than presently appreciated.

Notizen: Available evidence indicates that many of the antiprotozoal drugs currently in use significantly modify the immune response of the host. The effect depends on both the drug and the host. Some drugs enhance the immune response, some are immuno-suppressants, and others enhance some types of immune mechanisms while suppressing others. Future efforts in the development of antiprotozoal drugs should consider their effects on both the parasite and the immune response of the host. Also in the chemotherapy of protozoal infections consideration should be given to the combined usage of immunoenhancing agents and antiprotozoal drugs.

Notizen: A survey of protozoa polluting bottled mineral water in Mexico was carried out using samples obtained from the three best-selling brands of bottled mineral water in the country. The organisms were concentrated through filtration procedures and subsequently cultured in sterile media. The cultures were observed over four weeks, with identification to the level of genus and species. Most commonly found were the amoebae Naegleria gruberi, Acanthamoeba astronyxis, and Vahlkampfia vahlkampfi (trophic as well as cystic stages) plus one flagellate, Bodomorpha minima. No ciliates were detected. The public health importance of the findings is obvious, since some strains of Naegleria and Acanthamoeba have the potential to cause human disease that may lead to death.

Notizen: . Amphidinium carteri was unable to grow on nutrient-enriched seawater in the presence of 200 μg/ml fluoride (F) but could be adapted to grow successfully on this F concentration when repeatedly cultured with stepwise increases in sub-inhibitory F concentration. Electron microscopic investigation of the F-adapted dinoflagellate cells showed abnormal ultrastructural features in the chloroplast (especially the pyrenoid), mitochondria, and nucleus. Simultaneous comparison with the F-inhibited dinoflagellate cells showed that thylakoid formation was extremely disorganized by fluoride and that F-adaptation conferred a prolamellar-like configuration on the thylakoids in the center of the pyrenoid. This unexpected appearance of lamellae formation in the F-adapted cells suggested that the pyrenoid may be a center for thylakoid assembly. Such cells also showed large, intensely osmiophilic inclusions in the mitochondria. Microbodies are found in close juxtaposition to the mitochondria and chloroplast, suggesting an increased metabolic dependence on photorespiration. The F-adapted nucleus showed dark and light concentric rings in the nucleolus region, accompanied by other signs of mitotic activity, which were not observed in the F-inhibited cells. It was inferred that the F-adaptation may have required some form of genetic change resulting presumably in the development of a phenotype mutant.

Notizen: . The large fresh-water microaerobic amoeba Pelomyxa palustris does not contain mitochondria, but three types of bacterial endosymbionts are always present. Thus, it is of interest in the discussion of the possible origin of mitochondria from primitive prokaryotes. Gas exchanges (O2, CO2) and concentration of endosymbionts were determined in individual amoebae, in which the life cycle stage was noted. Grey type (stationary phase) amoebae had a lower O2 uptake and lower endosymbiont concentration than light type (growth phase) amoebae, and highest O2 uptake was found in centrifugal pieces of light type Pelomyxa, centrifuged in vivo, which contained nearly all of the endosymbionts. In light type amoebae, the respiratory activity was independent of O2 concentration between 1 and 21%, and, when compared on the basis of dry weight and protein, of the same order as that of other free-living protozoa. The R.Q. was slightly higher than 1, indicating that glycolysis does not play a significant role in energy metabolism. It is concluded that P. palustris is fully aerobic, and suggestions are presented as to the role of the endosymbionts in its respiratory metabolism.

Notizen: . A series of new in vitro systems for the cultivation of bloodstream forms of Trypanosoma (Trypanozoon) brucei brucei, T. (T.) b. rhodesiense, and T. (T.) b. gambiense was developed. The standard system consists of a feeder layer of fibroblast-like cells derived from embryos of New Zealand White rabbits (REF) or a mountain vole, Microtus montanus (MEF), with HEPES-buffered Minimum Essential Medium (MEM), with Earle's salts, supplemented with 15% inactivated rabbit serum. These two and other feeder layers were cross-checked with different sera to test for growth support of bloodstream forms of the three trypanosome subspecies studied. Cultures could be initiated with bloodstream forms from mammalian hosts or from cryopreserved stabilates. Metacyclic forms from infected Glossina m. morsitans could also be used as inoculum; they transformed within 6 h to bloodstream forms. Maintenance of cultures and growth properties are described in detail. Experiments were undertaken to confirm that the cultivated bloodstream forms still possess some of the characteristic features of pleomorphic bloodstream populations. Cultivated bloodstream forms were always infective for mice, and a surface coat could be demonstrated by electron microscopy. They could also be cyclically transmitted through tsetse flies, and the metacyclic forms from these flies could be brought back into culture. In vitro cloning with single bloodstream forms and metacyclic forms could be achieved with high cloning efficiency. The consumption of glucose and the production of pyruvate and lactate were determined.

Notizen: Fathead minnows, Pimephales promelas, raised from eggs in the laboratory, were experimentally infected with oocysts of Eimeria iroquoina from either P. promelas or the common shiner, Notropis cornutus. Within intestinal epithelial cells, trophozoites thought to be derived from the sporozoites contained a prominent electron-dense refractile body. Merozoites dedifferentiated into trophic forms by losing components of their apical complex and pellicle. The inner membrane components of the pellicle appeared discontinuous, and the micronemes became enclosed within vacuoles. Prior to merozoite formation, multinucleate meronts were limited by a single membrane. Golgi complexes were associated with the nuclei of this stage. Merozoites were formed by ectomerogony in one generation and by endomerogony in the final generation. In both forms of merogony the final nuclear division was coupled with the onset of differentiation of the merozoites and featured eccentric mitotic spindles associated with centrocones located within the nuclear envelope and with the precursors of the apical complex. A Golgi complex was closely associated with the nucleus and apical tip of the forming merozoite. Unlike other Eimeria species, the complete pellicle of the merozoites of the final asexual generation of E. iroquoina was formed within the cytoplasm of the meront, without association with the limiting membrane, thus, all pellicular components are synthesized de novo. The inner membranes of the pellicle initially appeared as longitudinal strips, each of which was associated with a pair of the 22–24 subpellicular microtubules. Mature meronts of the final asexual generation averaged 9 μm in diameter and produced 13–16 merozoites. With the exception of the internal completion of the pellicle of the final generation merozoites, the basic processes of merogony in fish Eimeria species are similar to those recorded in terrestrial hosts.

Notizen: The glycerophospholipids of the protozoon Tetrahymena pyriformis W are unique in that the polyunsaturated fatty acid γ-linolenate (18:3Δ6,9,12) is a major component of both the sn-C-1 and sn-C-2 positions. Tetrahymena were incubated with [1-14C]γ-linolenate. The positional distribution of the radiolabeled fatty acid in the three major glycerophospholipids was determined. [1-14C]γ-linolenate was found at both carbons of the three lipids, in general agreement with the mass distribution of γ-linolenate, except for markedly greater labeling at the sn-C-2 position of phosphatidylcholine. We hypothesize that an acyltransferase exists in Tetrahymena that can esterify γ-linolenate at both carbons during glycerophospholipid biosynthesis.

Notizen: Promastigotes of Leismania donovani cultured for either 3 or 10 days in vitro and inoculated intracardially into golden hamsters with an equal number of organisms from either population showed a 7-fold difference in infectivity when compared at both 10 to 16 days post-infection. Reproducible histochemical staining for the promastigote enzymes glucose-6-phosphate dehydrogenase (G6PDH) and peptidase after polyacrylamide gel electrophoresis showed two isoelectric variants of G6PDH (Bands 1 and 2) that displayed a 45% decrease (Band 1) and a 60% increase (Band 2) in total activity when 3- and 10-day-old promastigores were compared. Peptidase activity, present in a single band, increased 7-fold in 10-day-old promastigotes. A decrease in the lectin-induced agglutination of promastigotes by castor bean agglutinin (RCA60), specific for D-galactose and N-acetyl-D-galactosamine, was seen when 3- and 10-day-old promastigotes are compared. Antisera raised against sonicated 10-day-old promastigotes showed a unique precipitin band between the antiserum and sonicated 10-day-old promastigotes not found between the antiserum and sonicated 3-day-old promastigotes.

Notizen: The compound 6-(L-erythro-1,2′,3′-trihydroxypropyl)pterin, at a concentration of 50 pg/ml (“L-erythro-neopteria”), supports half-maximal growth of Crithidia fasciculata; biopterin at a concentration of 30 pg/ml is shown to yield similar growth. N2-dimethyl-6-(L-erythro-1′,2′,3′-trihydroxypropyl)pterin (A) was inactive even at 100 ng/ml. Synergism was observed with the N2-dimethylamino derivative (A) in the presence of suboptimal biopterin, its activity then being of the order of L-erythro-neopterin. In contrast, the stereoisomeric N2-dimethyl-6-(D-erythro-1′,2′,3′-trihydroxypropyl)pterin (“dimethyl-D-erythro-neopterin”) and its 3′-mono-phosphate only slightly enhanced growth under similar conditions but both threo-isomers had no supplementary activity. Biopterin-induced growth was slowed by 6-(D-erythro1′,2′,3′-trihydroxypropyl)pterin (D-neopterin); the threo-isomers had no such effect. An adaptive demethylation capacity by growing cultures and competition of biopterin uptake by D-neopterin seems likely. The report of the occurrence in Euglena of N2-dimethyl-6-(L-threo-1′,2′,3′-trihydroxypropyl)pterin and its 3′-mono-phosphate adds further interest to our observations.

Notizen: We investigated the macronuclear DNA genomes of several marine and fresh-water ciliates. The marine forms studied were: Uronema nigricans, Parauronema virginianum, Parauronema acutum, and two strains of Miamiensis avidus; the fresh-water ciliates included: Tetrahymena pyriformis, Paramecium octaurelia, and P. caudatum. The organisms were cultured axenically and the DNA extracted from isolated and purified macronuclear preparations. Reassociation rate constants of purified DNA preparations used to calculate kinetic complexity were determined both optically and by hydroxyapatite chromatography. Analytical complexity was determined chemically. Ciliate macronuclear DNA appeared to reassociate as a single unique sequence, except for a small fraction (4% of the total DNA) that was repetitive and renatured rapidly. Values for the kinetic complexities of macronuclear DNA in these forms varied over a relatively narrow range, from 1.5 to 3.8 times 1010 daltons, and were only 7–15x larger than that of the bacterium Escherichia coli. On the other hand, values for analytical complexities of macronuclear DNA of marine and fresh-water ciliates varied over two orders of magnitude and were related to the size of the animals. It is suggested that ploidy levels of macronuclear DNA in these ciliates may represent a functionally permanent amplification of the genome.

Notizen: Variations in DNA levels for log-phase Euglena have been shown to be directly related to the pH of the culture medium. Data are presented here for cultures grown in Cramer and Myer's medium at pH 3.0 and 6.8. DNA levels are ∼50% greater for Euglena at the higher pH value, confirming earlier reports.

Notizen: Guanylate cyclase activity decreased during the division phase of heat-shock synchronized Tetrahymena pyriformis, strain GL. However, when Ca2+ was removed by EGTA to negate the effects of the Ca2+-binding protein (calmodulin), which is required for the full activity of guanylate cyclase in this organism, no significant change in the enzymatic activity was observed throughout the cell cycle. On the other hand, the reduced guanylate cyclase activity at division phase was associated with a decreased level of calmodulin content. These results suggest that fluctuations in guanylate cyclase activity during the cell cycle would be dependent on the concentration of calmodulin.

Notizen: Identification and characterization of kDNA is described in the naturally occurring totally dyskinetoplastic species Trypanosoma equinum. Fluorescence microscopy of live cells, using the highly sensitive and specific probe DAPI (4,6,-diamidino-2-phenyl-indole), showed the presence of a diversity of extranuclear fluorescent bodies scattered along the length of the organism. Transmission electron microscopic studies revealed a close similarity between the distribution of these DAPI-fluorescing particles and of dense aggregates of nonfibrillar material resembling the kDNA of dyskinetoplastic strains of other species. Variable sized remnants of kDNA, occurring singly or in clusters, were found scattered throughout the mitochondrion. Analytical cesium chloride ultracentrifugation of total cellular DNA extracts showed a kDNA banding profile at a buoyant density equal to 1.691 gm/cm3, representing approximately 11% of the total cellular DNA content. Molecular spreads of isolated kDNA revealed a population of open circular molecules ranging in contour length from 0.11–9.69 μm.

Notizen: Morphogenesis in Conchophthirus curtus has been investigated by the use of protargol silver impregnation supplemented by selective use of scanning electron microscopy. Events are assigned to nine sequential stages; the last, stage 9, which involves maturation of the somatic ciliature and infraciliature as well as the final development of the buccal cavities of both proter and opisthe, occurs following cytokinesis. Stomatogenesis involves both the parental haplokinety and the deep kinetosomal unit (DKU). In perhaps a unique phenomenon, phylogenetically, the ciliated parental haplokinety forms the early oral primordium of the opisthe and is replaced in the proter by the DKU. The DKU then acts as a formative center for new kinetosomes that migrate into the developing opisthe primordium. Late in the process, the haplokinetal primordia of both proter and opisthe give rise to their respective DKU's. Some events of nuclear activity and somatic development are also described. The high degree of structural differentiation of this ciliate has provided the opportunity to examine temporal relationships during morphogenesis. We have found that somatic, buccal, and nuclear events proceed in a tightly coupled sequence. Hence, somatic and nuclear development can be directly correlated with the nine stages of stomatogenesis.

Notizen: The interaction of Leishmania with lysosomes within macrophages in vivo has been investigated. Lysosomes labeled with colloidal gold in vivo fused with phagocytic vacuoles containing Leishmania amastigotes within the macrophages of infected footpad tissue of BALB/c mice. This localization of Leishmania within macrophage phagolysosomes in vivo is the first confirmation for any obligate intracellulaire protozoon that parasite-lysosome interactions in vitro occur in vivo.

Notizen: The life cycle of Caryospora bubonis was studied in the Great Horned Owl. Owls fed sporulated oocysts became patent on days 8–13, and peak oocyst production occurred between days 14 and 18. Owls fed infected mice became patent on days 8–10, and peak oocyst production occurred between days 9 and 13. The reduction in length of the life cycle in owls fed infected mice provides evidence that both indirect and direct life cycles can occur in this species and suggests that parallels exist in the lives of various isosporid, eimeriid, and other coccidia that may not have been sufficiently appreciated in the past.

Notizen: Species of Eimeria occurring in fishes show important differences, especially in their life cycles, with respect to what is known for those forms found in mammals or other hosts. In some cases, at least, some small invertebrate may play the role of vector or intermediate host. Present studies of fish coccidia may throw light on the possible evolution of certain aspects of host-parasite interrelationships involving eimerian species in particular: for example, the earliest site (perhaps not the gut) of infection and even the “original” host group (perhaps invertebrates rather than vertebrates).

Notizen: Problems of the taxonomy and, especially, of the nomenclature of various species of coccidia of direct importance to humans are further discussed. The diversity of host potential and site selection is also mentioned, with the prediction that, hopefully, our knowledge will be much greater 15 years from now than it presently is.

Notizen: The use of drugs in domestic animals is dominated by considerations of cost-effectiveness and profitability. They are extensively used against coccidial infections of poultry where they are an important factor in intensive husbandry. Ionophore antibiotics, which dominate this field, may have applications in ruminants. Imidocarb is of therapeutic and prophylactic value against babesial infections and there are new prospects for control of theileriasis. Effective drugs for the control of African trypanosomiasis are limited and attention is being given to alternative uses of available compounds.

Notizen: Histological examination of gills of the rock crab (Cancer irroratus Say, 1917) taken from specimens collected in polluted and nonpolluted areas indicated that gill discoloration was not directly related to the degree of microbial fouling. Light microscope studies demonstrated that bacteria, peritrich and suctorian ciliates, and some naviculoid diatoms were the principal members of the fouling community. Examination of the bacteria by electron microscopy showed that several distinct types were present, and that some of these were attached to the gill by holdfast structures while others were enmeshed in a filamentous slime layer. An unusual small marine flagellate and an unidentified amoeba were also examined ultrastructurally.

Notizen: The endogenous development of Eimeria christenseni was studied in 10 two- to four-week-old kids inoculated with 106-107 sporulated oocysts. They were killed at intervals from two to 26 days after inoculation, and their tissues were examined for endogenous stages of the coccidian by light microscopy. Such stages were found in the small intestine and mesenteric lymph nodes. In the sexual cycle, two generations of meronts were found. The first generation developed in endothelial cells of lacteals in the jejunun and ileum and mesenteric lymph nodes, and mature meronts were first seen 14 days after inoculation. The second generation developed in epithelial cells of the glands of Lieberkuehn in the jejunum and ileum and in mesenteric lymph nodes, and its mature meronts were first seen by 16 days. Sexual stages were present mostly in epithelial cells of the tips and sides of the villi and less frequently in crypt cells of the jejunum and ileum. Mature macrogametes and microgamonts and oocysts were also first seen by 16 days. The prepatent period was 17 (14-23) days; the patent ranged from 8 to more than 30 days. Sporulation time was 3-4 days at 30°C. E. christenseni was found to be pathogenic, kids inoculated with 1-5 × 105 sporulated oocysts exhibited the following signs: severe diarrhea, anorexia, polydipsia, poor hair coat, and extreme weakness. They recovered about a month later, but their growth rates appeared to be lower than those of uninoculated animals kept under the same conditions. One kid died 20 days after inoculation with 107 oocysts.

Notizen: The ultrastructure of Rhodomonas lacustris was examined using scanning and transmission electron microscopy, and the pigments were scanned in vivo by the opal plate technique. The periplast is composed of hexagonal plates, with trichocysts at the corners. This cryptomonad lacks a gullet but has a deep ventral furrow, in which more than 20 large ejectosomes are located. The chloroplast has mainly three thylakoids per lamella, and it is structurally reminiscent of that of certain haptophycean flagellates. The pigments appear to be as usual within the photosynthetic cryptomonads, with phycoerythrin type PE I (phycocyanin was not detected). Other cellular inclusions (e.g., lysosome-like vesicles and the contractile vacuolar complex) are also discussed.

Notizen: Following conjugation in ciliates, the usual fate of the old pre-conjugant macronucleus is resorption. In some species, however, old macronuclei, or their fragments, have the ability to reform functional vegetative macronuclei when new macronuclear anlagen are defective. The present work on Euplotes shows that if anlagen are allowed to carry out their essential roles in early exconjugant development, including influence on cortical reorganization such that feeding can resume, they can then be permanently damaged by UV-microbeam irradiation and regeneration of old macronuclear fragments can occur. E. aediculatus exconjugants were anlage-irradiated at 40–60 hr of development and the irradiated cells cultured individually and fed. Squashes revealed enlargement and anteriorward migration of the persistent (posterior) macronuclear fragments. The first post-conjugant fission of such cells was delayed (times ranged 6–43 days) and did not seem to involve the damaged anlagen, which remained rudimentary, did not divide along with the cells, and were subsequently resorbed. It appeared that cell fission was supported by the fragments of the old macronuclei, which either divided or partitioned themselves between the two daughter cells. Mating tests performed on early clones derived from irradiated exconjugants revealed ample conjugation competence; intraclonal conjugation in such clones was also apparent. The absence of the immature period seen in normal exconjugants provides further evidence that the clones arose from cells with regenerated macronuclei.

Notizen: Cell free preparations of Tetrahymena thermophila contain an enzyme that catalyzes the direct desaturation of stearoyl CoA to octadecenoic acid. The enzyme is associated with the microsomal fraction of the ciliate. Substrate for the enzyme consists of either free stearic acid or stearoyl CoA. Both ATP and CoA are required when free stearate is the substrate and are also highly stimulatory when stearoyl CoA is the substrate. With stearoyl CoA as the substrate, either NADH or NADPH are required for desaturase activity. In the presence of ATP and CoA, either NAD or NADP can replace NADH and NADPH. Desaturase activity is optimal when the enzyme is incubated at a pH of 7.2 and a temperature of 30–35°C. Highest levels of the stearoyl CoA desaturase are found in stationary phase ciliates grown at 35°C.

Notizen: SYNOPSIS. The name Nebela tuberculata comb. nov. is proposed for the testate ameba formerly known as Difflugia tuberculata (Wallich, 1864) Archer, 1867. The generic reassignment is based upon the test construction material which is shown, by electronmicrographs and X-ray diffraction, to be autogenous regular and irregular silicious rods. A brief history of the animal is recounted utilizing works of several authors who have encountered the ameba over the last century.

Notizen: SYNOPSIS. The ultrastructure of the mitochondrion during transformation of Leishmania donovani from amastigote to promastigote stages was studied by morphometric analysis. There was a slight but significant decrease in relative mitochondrial volume (Vvli) during transformation. This decrease was linear with time for at least 27 hr. No change in relative lipid inclusion volume (Vvli) was observed during transformation. The ratio of inner to outer mitochondrial surface membrane densities (Svmii:Svmio) also remained unchanged. Although the relative number of cristae remains unchanged after transformation, there is an increase in cristae number in the portion of the kinetoplast opposite the flagellum.

Notizen: SYNOPSIS. Electron-dense deposits indicating possible Ca-binding sites were found at the ciliary base of Paramecium caudatum fixed in a glutaraldehyde solution containing 5 mM CaCl2. The deposits appeared mainly at the inner surface of the ciliary membrane above the “ciliary necklace” region, although they could also occur in the space between the outer and the central microtubules. In some cases a ring of exactly 9 deposits was found in a ciliary cross section of a cilium.

Notizen: L'infraciliature ventrale est presque entièrement détruite lors de la conjugaison: la totalité des structures buccales, tous les cirres fronto-ventraux et transversaux, certains cirres caudaux. Le retour à l'état végétatif s'effectue en 2 étapes. Première Etape.–Différenciation d'un nouvel ensemble de cirres, dépourvu du cirre 1/I (selon la nomenclature de Wallengren); néoformation, à partir d'une ébauche située sur le territoire présomptif du cirre manquant, d'une partie des structures buccales (la moitié antérieure de la frange adorale de membranelles). Deuxième Etape.–Remplacement de tous les cirres par un nouvel ensemble comportant le cirre 1/I; différenciation des structures buccales manquantes (moitié postérieure de la frange adorale, ciliature parorale)Les rapports morphogénétiques entre le cirre 1/I et la ciliature parorale suggèrent que le territoire de ce “cirre paroral” est homologue des territoires stomatogènes d'autres Hypotriches tels que Kahliella et Stylonychia.SYNOPSIS. During conjugation of Euplotes, the ventral ciliature, including the entire oral apparatus, all the fronto-ventral and transverse cirri, and some of the caudal cirri, is nearly completely lost. As followed in silver-stained preparations, the redifferentiation of the ciliature proceeds in 2 steps. The first step entails differentiation of a new complement of cirri, except for cirrus 1/I (according to the nomenclature of Wallengren), and neoformation of the anterior part of the adoral zone of membranelles (AZM) from a primordium located in an area that would be expected to give rise to the missing 1/I cirrus. The 2nd step involves replacement of all the cirri, including 1/I, and completion of the oral apparatus by redifferentiation of the posterior half of the AZM and of the paroral ciliature. The spatial morphogenetic relationships between cirrus 1/I and the paroral ciliature suggest that the area of this “paroral cirrus” is homologous with the stomatogenic areas of other Hypotrichida, such as Kahliella and Stylonychia.

Notizen: SYNOPSIS. Material from various steps obtained in the French pressure cell technic of preparing antigen from Plasmodium knowlesi-infected red cells, was examined by electron microscopy. A positively charged colloidal iron solution was used to differentiate between membranes of host red cells and parasites. Red cell membranes take the stain, whereas parasite membranes do not. This antigen which has been used previously to protect monkeys against P. knowlesi appears to consist almost entirely of membrane-bounded vesicles. Some of these vesicles contain a fine granular material, whereas others appear empty. The antigen failed to stain with the positively charged iron solution, which suggests that it is free of contamination by host cell membrane.

Notizen: SYNOPSIS. The glucose transport system in Leishmania tropica promastigotes was characterized by the use of labeled 2-deoxy-D-glucose (2-DOG), a nonmetabolizable glucose analog. The uptake system has a Q10 of 2 and a heat of activation of 10.2 kcal/mole. The glucose transport system is subject to competitive inhibition by 2-DOG, glucosamine, N-acetyl glucosamine, mannose, galactose, and fructose which suggests that substitutions in the hexose chain at carbons 2 and 4 do not affect carrier specificity. In contrast, changes at carbon 1 (α-methyl-D-glucoside, 1,5-anhydroglucitol) and carbon 3 (3–0-methyl glucose) lead to loss of carrier affinity since these sugars do not compete for the glucose carrier. Sugars that compete with the glucose carrier have one common feature—they all exist in the pyranose form in solution. The carrier for D-glucose does not interact with L-glucose or any of the pentose sugars tested. Uptake of 2-DOG is inhibited by glycerol. This inhibition, however, is noncompetitive; it is evident, therefore, that glucose and glycerol do not compete for the same carrier. Glycerol does not repress the glucose carrier since cells grown in presence of glycerol transport the sugar normally.

Notizen: SYNOPSIS. Interaction of several plant lectins with the ciliates Stylonychia mytilus, Euplotes aediculatus, and Tetrahymena pyriformis GL, was investigated. The motility of Stylonychia is specifically inhibited by treatment with concanavalin A, with which the 2 other ciliates react only weakly. Stylonychia can regain its motility by shedding the lectin-loaded surface components and rebuilding a new pellicle. Other lectins used in this study did not interact with these ciliates.

Notizen: SYNOPSIS. Crithidia harmosa and Crithidia fasciculata were compared immunologically by the indirect fluorescent antibody (FA) method and by agglutination. The FA technic yielded more specific results with cellular-debris than with whole-cell antigens. The immune sera collected from chickens 9 days after the last inoculations with whole-cell antigens had higher homologous titers than those collected after 4 days. Major antigenic differences between the 2 species were revealed by both methods. The electrophoretic patterns of C. harmosa and C. fasciculata obtained by polyacrylamide gel slab electrophoresis differed in the numbers and relative mobilities of their component bands.

Notizen: SYNOPSIS. Bacteria can be eliminated from protozoan cultures by adding a bacterium supplanting those difficult to destroy, if the added bacterium can be easily eliminated. Pseudocohnilembus persalinus was cultivated axenically in a simple medium containing peptones, nucleic acid extract, and vitamins. Growth was stimulated by saturated or unsaturated fatty acids.

Notizen: SYNOPSIS. Acanthamoeba castellanii grows in a minimal medium (AMLIV) containing only arginine, methionine, leucine, isoleucine, and valine as sole nitrogen sources, other than vitamins, when glucose is the carbon source. With acetate as the carbon source, glycine must be added to AMLIV. Doubling time in AMLIV varies according to the ratio of amino acids concentrations. Several combinations yield Td values of ∼ 70 hr.

Notizen: Book reviewed in this article:Garcia, L. S. & Ash, L. R. 1975. Diagnostic Parasitology: Clinical Laboratory Manual

Notizen: SYNOPSIS. Tetrahymena grown overnight in deep cultures were incubated for 1 hr with [1-14C]labeled substrates in the presence or absence of 3-mercaptopicolinic acid (3-MPA). 3-MPA inhibited appearance of label in glycogen from bicarbonate, acetate, pentanoate, octanoate, and succinate, but not from glycerol or glucose. In vitro assays of phosphoenolpyruvate carboxylase and phosphoenolpyruvate carboxykinase activity showed that both enzymes were about equally distributed between the particulate and cytosol fractions. 3-MPA inhibited phosphoenolpyruvate carboxykinase from both the cytoplasmic and particulate fractions, but had no effect on phosphoenolpyruvate carboxylase from either location. These results suggest that the in vivo effects of this drug are due to inhibition of glyconeogenesis at this site.

Notizen: SYNOPSIS. It was demonstrated with the aid of light and electron microscope studies that Sorites marginalis (Lamarck) harbors zooxanthellae. The hosts were scraped from Thalassia testudinum Konig growing in Key Largo Sound (Florida, U.S.A.) and immediately preserved in appropriate fixatives. Zooxanthellae were distributed unevenly throughout all the chamberlets; only a few symbiotes were found in the embryonic chambers and the inner or outer chambers, but the intermediate chambers were packed with symbiotes. The outer chambers contained many food vacuoles in addition to symbiotes. Some zooxanthellae might have been in the process of degeneration or digestion. The symbiotes were found to have a typical dinoflagellate nucleus, a single large lobate cortical chloroplast with one stalked pyrenoid, an accumulation body, and many starch granules. The nonmotile stage of the zooxanthella was similar, but perhaps not identical, to Symbiodinium microadriaticum Freudenthal from various hosts.The foraminiferan host is heterokaryotic with hundreds of generative (small) nuclei and scores of vegetative (large) nuclei. Most of the generative nuclei were found in the embryonic apparatus and the inner chambers. Most of the vegetative nuclei were found in the inner and outer chambers.

Notizen: SYNOPSIS. The effects of fixation with various concentrations of glutaraldehyde or formaldehyde, acetone or ethanol, and freeze-drying on 5 phosphatases of Eimeria tenella and chick kidney cell cultures were demonstrated in situ. Gultaraldehyde inactivated the phosphatases more than did the formaldehyde, but the effect of the combination of the 2 (Karnovsky's fixative) was greater than that of either glutaraldehyde or formaldehyde alone. The higher the concentration of aldehyde and the longer the duration of exposure, the greater the inactivation. The order of sensitivity to aldehyde fixation of the enzymes tested was glucose-6-phosphatase 〉 thiamine pyrophosphatase 〉 5′-nucleotidase 〉 adenosine triphosphatase 〉 acid phosphatase. Cytologic detail was preserved more efficiently with glutaraldehyde than with formaldehyde. Optimal preservation of enzyme activity for cytochemistry was with 2% glutaraldehyde for 30 min or 2% formaldehyde for 1 hr for G-6-Pase. TPPase, and 5′-nucleotidase, and with 2% glutaraldehyde or 2% formaldehyde for 2 hr with ATPase and AcPase.Quenching with subsequent fixation in cold acetone or ethanol resulted in complete inactivation of G-6-Pase, TPPase, and 5′-nucleotidase; although cells fixed in this manner yielded large amounts of reaction product for ATPase and AcPase, the distribution was diffuse, and some of it appeared to be artifactual. Quenching with subsequent freeze-drying was unsatisfactory because nearly all of the cell layers rolled off the cover glasses.

Notizen: SYNOPSIS. The behavior of the contractile vacuole of Tetrahymena pyriformis W has been recorded and analyzed quantitatively by cinephotography. The vacuole fills in a stepwise fashion by the confluence of ampullae which appear regularly at the beginning of systole and whose membranes are continuous with that of the contractile vacuole throughout the cycle. The vacuole may subsequently fill slowly by a means not discernible by light microscopy. The vacuole rounds up at the beginning of systole and shortly thereafter the ampullae reappear around the perimeter of the vacuole. They are expanded by fluid forced into them from the vacuole. Round-up and the mode of growth of the ampullae indicate that the contractile vacuole is truly contractile. Expulsion occurs soon after the appearance of the ampullae and terminates the cycle. Contraction is initiated at regular intervals by a timing mechanism which is independent of the size of the vacuole. Suitable terminology to describe the structure and behavior of the contractile vacuole is discussed.

Notizen: SYNOPSIS. Cells of Euglena gracilis strain Z were extracted with trichloroacetic acid. Samples of gross cellular protein were hydrolyzed by a variety of reagents. Amino acids released by these procedures were analyzed and the overall composition of cell protein was quantitatively determined.

Notizen: SYNOPSIS. Pellicles of the ciliate Tetrahymena pyriformis strain GL (phenoset A) were isolated by a new procedure. Oral apparatuses were also purified by a modification of a previous method. Both preparations were characterized by electron microscopy.Proteins of the isolates were separated by analytical SDS polyacrylamide gel electrophoresis. The isolated pellicles, which included oral apparatuses, contained only 6 major proteins (gel bands), designated A through F. Bands A, B, and C, were found in the pellicle fraction, but not in the oral apparatus fraction. Therefore, these proteins are believed to be present in the somatic cortex of Tetrahymena. Bands D and E were greatly enriched in the oral apparatus fraction; these proteins are therefore believed to be present primarily in the oral apparatus. Band F, identified as tubulin, was present in both preparations. Molecular weight determinations and some selective solubilization experiments are also presented.

Notizen: SYNOPSIS. Uridine uptake was examined in Tetrahymena pyriformis GL-7 in defined medium under conditions where food vacuole formation is not a significant factor in solute acquisition by the cell. The results indicate the presence of a saturable mechanism which follows Michaelis-Menten kinetics. When corrected for diffusion the apparent Km for the carrier is 2.3 ± 0.6 μM and the Vmax is 7.3 ± 0.2 × 10−7 nmoles/cell/min. It is evident from nucleotide pool analysis that most of the radioactivity of externally supplied [3H]uridine appears in UMP with the remainder in UTP. Uridine is apparently phosphorylated immediately upon entry into the cell and neither uridine-cytidine kinase activity nor RNA synthesis are rate-limiting in the uptake process. Uridine transport is competitively inhibited by a variety of ribo- and deoxyribonucleosides as well as several nucleoside analogs. Neither uracil nor ribose or deoxyribose are effective inhibitors of uridine transport indicating the carrier is specific for the nucleoside. There is little difference between the Ki values for ribo- as opposed to deoxyribonucleosides except in the case of deoxyguanosine which is much less effective as an inhibitor under the conditions of this study, than all the other nucleosides, including guanosine.

Notizen: Book reviewed in this article:Canning, E. U. 1975. The Microsporidian Parasites of Platyhelminthes: Their Morphology, Development, Transmission and Pathogenicity

Notizen: SYNOPSIS. Nuclear features of dinoflagellates that were used originally to support the Mesocaryota concept are reviewed. The fibrillar diameter of dinoflagellate chromatin, low level of chromosomal basic proteins, membrane attachment of chromosomes and swirl pattern observed in sectioned chromosomes are features that support a prokaryotic affinity. The presence of repeated and highly complex DNA, a S-phase of DNA synthesis in the cell cycle, presence of basic proteins, and the reinterpretation of extranuclear microtubules as a spindle support the contention that dinoflagellates are eukaryotes. This combination of prokaryotic and eukaryotic features suggests that dinoflagellates are a geologically old group and that perhaps they diverged from the higher eukaryotic lineage before evolution of eukaryotic chromatin but after the evolution of repeated DNA. The 2 patterns of carotenoid composition exemplified by the presence of peridinin or fucoxanthin suggest separate origins of dinoflagellate plastids, perhaps by prokaryotic and eukaryotic capture. It is suggested that the species possessing fucoxanthin obtained their plastids by capture of photosynthetic eukaryotes.A new class and order, Syndiniophyceae and Syndiniales, are proposed for the dinoflagellates with low chromosome numbers, V-shaped chromosomes, chromosomes containing a sufficient quantity of basic proteins detectable histochemically, possession of centrioles associated with mitosis, intracellular parasitism as a mode of nutrition, and lack of a cellular covering containing plates.Ultrastructural and paleontologic evidence indicates that the thecate is more primitive than the nonthecate condition.The Prorocentrales are considered to be primitive and their thecal construction is reinterpreted as having epithecal and hypothecal regions surrounding a flagellar pore region containing 7 plates. Acritarchs resemble cysts of modern dinoflagellates in size, structure, and chemical composition except for the absence of a polygonal excystment aperture and lack of any indication of transverse and longitudinal flagellar grooves on the acritarchs. The suggestion that some acritarchs may have dinoflagellate affinities is supported by the occurrence of modern dinoflagellates (Prorocentrales) which lack a theca of numerous polygonal plates and lack transverse and longitudinal flagellar arrangement. The Prorocentrales, as opposed to the more typical Dinophyceae, perhaps represent the type of organism that produced some acritarchs.

Notizen: SYNOPSIS. Progress in ciliatology and in allied fields may demystify ciliate phylogenetics. Concentration on hymenostomes (mainly Tetrahymena and Paramecium) may have obscured directional features of ciliate physiology in phylogenetic problems. Therefore, means are suggested for “domesticating” the presumptively primitive, predominantly marine, sand-dwelling gymnostomes having nondividing diploid macronuclei. The prize quarry is the marine psammophile Stephanopogon whose homokaryotic condition may mark it as a living fossil. Eventual axenic cultivation of these “primitive” ciliates may be aided by use as food of easily grown photosynthetic prokaryotes, some isolated from the marine sulfuretum or adjacent aerobic muds and sands where “karyorelictid” ciliates flourish.We assume that: (a) the macronucleus evolved as a coordinator of chemical and physical signals, for efficient detection of food and toxins; (b) oral structures evolved meanwhile as sensors as well as mechanical food-gatherers. This conjunction enabled complexity of adaptive behavior and evolutionary success. Ciliate origins cannot be considered apart from origin(s) of phagotrophy and its underlying versatile heterotrophy. Because of the well developed heterotrophy in some photosynthetic prokaryotes (including several proposed as food organisms), they are viewed as alternatives to blue-green algae as forebears of eukaryotes. Nor can ciliate origins be considered apart from origin(s) of eukaryotes. A check of these assumptions—that Stephanopogon and gymnostomes with nondividing macronuclei are primitive—may be forthcoming from sequencing amino acids in certain key enzymes, given an adequate sampling of ciliates, flagellates (especially dinoflagellates and cryptomonads), lower fungi, and photosynthetic prokaryotes other than blue-green algae.

Notizen: SYNOPSIS. The sensitivity of Paramecium tetraurelia (=P. aurelia syngen 4) cells to pulse treatments with various doses of Actinomycin D (AMD) was estimated by comparing the generation times of treated and untreated sister cells. It was found that the delay of division in treated cells depended on the concentration of AMD, on their “age” at the time of the pulse treatment, and on their individual sensitivity.Sensitivity of Paramecium to AMD changes during the cell cycle in a predictable way. About 3 1/2 hr before the normally expected cell fission (total generation time ∼ 5 1/2 hr) there is a decrease of sensitivity. Thereafter, the cell enters a new stage with a progressive increase of sensitivity. This 2nd phase ends at the “transition point” (∼ 2 hr before cell division), when sensitivity drops abruptly. The division process itself may be altered and slowed down by high concentrations of AMD, even if the drug is applied after the transition point, but this process can never be completely annulled. The impairment of the division mechanism may lead to morphologic anomalies in the offspring.Resorption of oral anlagen in P. tetraurelia probably never occurs during the cell cycle after AMD treatment.The reason for individual variability of the cells, mechanisms controlling development, and the question of an obligate sequence of gene action in each cell cycle are discussed.

Notizen: SYNOPSIS. Information relating to the ultrastructure of 4 organellar systems of flagellates—nuclei (including mitosis), flagella, mitochondria and chloroplasts—is examined for bearing on the probable phylogeny of the principal flagellate groups, first considered singly and then in combination. The mitotic mechanism has not proved to be as conservative a character as might be hoped, but still remains characteristic for the average condition in many of the groups. Flagellar features are useful if allowance is made for the reduction or multiplication of the basic pair, and the loss of lateral and terminal hairs seems to have occurred independently several times. The presence of paraxial rods within flagella may be a useful indication of affinity. Rootlet systems are not dealt with in detail here, although the possible similarity between axial microtubular sheets in axostylar flagellates and some members of the green algae containing “manchettes” is noted. The basic patterns of chloroplast internal structure are summarized and their general agreement with other characters is affirmed, noting however that cryptomonads may be closer to the green flagellates (including euglenoids) than is generally accepted. Attention is drawn to the potential value of internal mitochondrial morphology as an indicator of large assemblages. Finally, a “tree” based on multiple cell organizational features is presented and discussed.

Notizen: SYNOPSIS. By fine-structural studies of speculation stages in the European flat oyster pathogen Marteilia refringens this organism is shown to be a member of the protozoan class Haplosporea. This conclusion is based on observations that haplosporosomes are found in plasmodia and spores and that sporoplasm delimitation occurs by internal cleavage within plasmodia. During sporulation plasmodia become sporangiosori by enlarging, cleaving internally into ∼ 8 sporangia, each of which then cleaves internally into 3 or 4 spore primordia. The primordia divide into 3 uninucleate sporoplasms of graded sizes, the smallest lying in an eccentrically located vacuole of the medium-sized one, and the latter situated within an eccentric vacuole of the largest. Haplosporosomes are found only in the cytoplasm of the largest sporoplasm. Mature, living spores measure 3.9(3.5–4.5)μm diameter and are enclosed in a continuous wall without a mantle of epispore cytoplasm as in other Haplosporea.An amended genus and species description of Marteilia refringens is presented, based on light and electron microscope observations.

Notizen: SYNOPSIS. Phagocytosing Amoeba proteus at different stages of forming foodcups have been observed by scanning electron microscopy. A nonphagocytosing ameba is characterized by dorsal and lateral ridges running longitudinally over the posterior half of the cell and its attachment to the substrate over small areas. When stimulated by prey organisms, the ameba loses polarity and ridges, and adheres to the substrate more firmly over a wider area of contact. Then it forms broad pseudopods to surround its prey and this results in the formation of foodcups. The surface of all amebae is covered with small projections, and membranous blebs are often seen on the surface of phagocytosing organisms.

Notizen: SYNOPSIS. The life cycles of 3 strains of Allogromia laticollaris, a monothalamous foraminiferan, have been studied. Each of the strains had a different, nonclassical, and basically apogamic, life cycle. The Cold Spring Harbor (CSH) strain regularly alternated between 2 agamontic forms: agamont I (uninucleate and diploid) and agamont II (multinucleate and diploid). The complete life cycle took 26 days. Sexual reproduction was rare (0.01%) and autogamous. Small numbers of organisms also underwent budding, binary fission, and cytotomy. The life cycles of the Towd Point (TPA) and Sippewissett (SIP) strains were comparatively abbreviated. Agamont II dominated their typical life cycles, which were completed in 16-18 days. The life cycle of SIP was basically a continuous cycling of the agamont II phase. Approximately 75% of the schizozoites of the TPA strain developed into agamont II. The other 25% alternated between agamont II and agamont I phase.In the CSH strain schizozoites with ∼ 8 (range 5-15) nuclei characterized newly formed agamonts II. More nuclei (∼ 25) were found in the other 2 strains. The nuclei in young agamonts II underwent rapid morphologic changes leading to a “mushroom-like” chromosome appearance and extensive RNA synthesis. Nucleolar material accumulated at the nuclear periphery and eventually was discharged to the cytoplasm. Karyokinesis took place without the breakdown of the nuclear membrane.The single nucleus of young agamont I forms was proportionally quite large. The S1 phase occurred quite early (2-5 days) in this part of the life cycle. RNA in the CSH strain formed a compact, subcortical, coarsely granular ring, while in the TPA it was cortical and differentiated into finely granular matrix with randomly distributed coarse granules. During the G2 phase the nucleus became further enlarged and eventually amoeba-form. Intermediate stages in nuclear breakdown were not found.