ALBERT

Notes: Kallinikova, V. D. 1977. Kletochnaya Organella Kinetoplast. [The Kinetoplast, a Cell Organelle (in Russian).]

Notes: McClure, H. E., Poonswad, P., Greiner, E. G. Laird, M. 1978. Haematozoa in the Birds of Eastern and Southern Asia.

Notes: Long, P. L., Boorman, K. N. Freeman, B. M. 1978. Avian Coccidiosis. Poultry Science Symposium #13.

Notes: The effect of exposure period and concentration of algae on the frequency of infection of aposymbiotic ciliates by algae obtained from the same clone of Paramecium bursaria syngen 2, was studied. The frequency of infection was roughly proportional to the algal concentration and to the exposure time of ciliates to algae. The relationship of algal concentration to infection frequency closely fitted the Poisson distribution curve for N = 1, suggesting that the minimum number of algae required to infect a single ciliate is 1. However, the data also strongly suggested that the average number of algae required to initiate infection of an average ciliate was ˜ 1,000. Three possible resolutions of this situation are: (a) the selection by the ciliate of a rare infective variant from a heterogeneous population: (b) the rare escape of an alga from digestion by the ciliate; and (c) the requirement for a large number of algae-ciliate contacts to induce susceptibility in the ciliate. Splitting the exposure of ciliates to algae into 2 periods of 0.5 h, separated by 5 h in the absence of algae, produced a much higher frequency of infection than a single l-h exposure, supporting the suggestion that the large number of algae is required to induce susceptibility in the ciliate which can then be infected by as few as a single algal cell.

Notes: A plaque technic was evaluated and used for the isolation and enumeration of small, free-living amebae in lake-bottom samples which were collected each month for one year from a Nebraska lake. Several culture media were evaluated, and a simple glucose-salts medium was chosen. The most frequent ameba in the lake-bottom samples was Acanthamoeba polyhaga which underwent marked increases and decreases in population densities during the collection period. This pattern was not correlated with water temperature, bacterial counts, and nitrate or phosphate levels. Other species of amebae of the genera Hartmannella, Vahlkampfia, Naegleria, Paratetramitus and Echinamoeba were isolated. Most of these were either found infrequently or remained at relatively low, constant densities throughout the year. In addition, 4 species of Acrasieae of the genera Dictyostelium and Polysphondylium were isolated from the lake-bottom samples.

Notes: SYNOPSIS Hydroxyurea (HU) inhibits increase in cell number in cultures of Crithidia fasciculata. Complete inhibition is produced by 8 mM and higher concentrations. If HU is not removed, population growth resumes in 45–50 h: if HU is removed, partially synchronous growth occurs through 2 cycles. During HU inhibition, the rate of DNA synthesis is reduced to 1% of that in exponentially growing cultures; protein and RNA syntheses continue at slightly reduced rates. Mean cell size and protein and RNA contents per cell increase; rate of oxygen consumption per mg cell protein remains constant. The behavior of a culture upon addition of HU and upon its removal agrees with predictions based on the hypothesis that the only direct effect of HU is to block DNA synthesis. The synchrony produced by HU is judged satisfactory for investigations of kinetoplast and nuclear replication but not for biochemical characterization of other aspects of the cell cycle.

Notes: SYNOPSIS Dihydrofolate reductase (E.C. 1.5.1.3) from Plasmodium falciparum and from its host, the owl monkey (Aotus trivirgatus). were partially purified and characterized. The molecular weight of the parasite enzyme was estimated to be over 10 times as high as that of the host enzyme. The host enzyme had 2 pH optima whereas the parasite enzyme only one. The activity of the host enzyme was greatly stimulated by KCI and urea, while that of the parasite enzyme was inhibited at high concentrations of such chaotropic agents. Km of the parasite enzyme was significantly higher than that of the host enzyme. The parasite enzyme had much lower Ki for pyrimethamine than the host enzyme. Dihydrofolate reductases isolated from pyrimethamine-resistant and pyrimethaminesensitive strains of P. falciparum were found to be similar.

Notes: The chelator GEDTA was used to show that Ca2+ is required for the growth of Tetrahymena pyriformis strain W and for normal galvanotactic responses and swimming.

Notes: Entamoeba histolytica required CO2 for growth in axenic culture while growth was inhibited by H2. The organism was tolerant to 5% O2 in the gas phase and it was able to detoxify products of O2 reduction in the medium. The ameba did not require a negative oxidation-reduction potential for axenic growth. However, little or no free O2 was present in media exposed to 5% O2 in the gas phase. Growth was improved by adding yeast extract to the medium.

Notes: Twelve acid hydrolases, 4 near-neutral hydrolases and alkaline phosphatase were demonstrated in 0.34 M sucrose homogenates of Trypanosoma cruzi strain Y: p-nitrophenylphosphatase and α-naphthylphosphatase, with optimum pH at ˜ 6.0; α-galactosidase, β-galactosidase, β-glucosidase, N-acetyl-β-glucosaminidase, cathepsin A and peptidase I and III, with optimum pH between 5.0 and 6.0: and arylsulfatase cathepsin D, α-arabinase and α-mannosidase with optimum pH at ˜ 4.0 α-Glucosidase, gluccse-6-phosphatase and peptidase II had optimum pH at ˜ 7.0. β-Glycerophcsphatase had a broad pH-activity curve from 4.0 to 7.4, with maximum activity at pH 7.0. The main kinetic characteristics of these enzymes and their quantitative assay methods were studied.No activity was detected for α-fucosidase, β-xylosidase, β-glucuronidase, elaidate esterase. acid lipase, and alkaline phospho-diesterase.

Notes: SYNOPSIS. Cortical ultrastructure of the scuticociliates Dexiotricha media and Dexiotricha colpidiopsis was investigated. The following elements of the somatic cortex were studied: the cell membrane, alveolar membranes and the epiplasm, kinetodesmal fibers, postciliary and transverse microtubular ribbons, and transverse fibers associated with single and paired kinetosomes; mitochondria and single microtubules located in interkinetal ridges; mature and early extrusion stages of mucocysts: the expulsion vacuole pore and tube, the nephridioplasm and the cytoproct. In the buccal cortex, the paroral kinety-ribbed wall complex, the 3 polykineties, and the cytostome-cytopharynx were investigated.Comparative survey of ciliate ultrastructure indicates 2 principal orientation patterns for kinetodesmal and postciliary fibers, recognition of which leads to reevaluation of the theory of paroral kinety formation and the ideas of homology based on this theory. Ultrastructurally, the scuticociliates are not distinct from tetrahymenines and peniculines; the 3 groups appear to be 1 assemblage.

Notes: SYNOPSIS. In this study the characteristics of the synthesis of DNA and RNA in the nuclei of Loxodes were investigated. Loxodes striatus is a primitive ciliate with 2 pairs of structurally differentiated diploid nuclei, the macro- and micronuclei. The macronuclei are differentiated morphologically into a clearly recognizable central core and an outer zone.To determine DNA and RNA synthesis, individual organisms were analyzed by autoradiography after incubating groups of cells with a 3H-labeled precursor ([3H]thymidine for DNA and [3H]uridine for RNA).The following observations were made: (A) All portions of macro- and micronuclei appeared to contain DNA as judged by the localizations of incorporated [3H]thymidine. (B) The macro- and micronuclei did not synthesize DNA at the same time; moreover, the duration of DNA synthesis in the former was much longer than of the latter nucleus. (C) Replication of DNA in the inner core and outer zone of the macronucleus occurred at separate times with little if any overlap. (D) All of the detectable [3H]uridine incorporation was found in the macronucleus and none in the micronucleus. Within the macro-nucleus the central core was more heavily labeled. (E) The quantitative differences in the label of the different components of the nuclear complex were investigated. (F) Contrary to the previously reported information our results suggest that DNA synthesis can occur in adult macronuclei.The possible explanation of these results is discussed in the context of the nuclear evolution of ciliates and of recent information on nuclear differentiation.

Notes: SYNOPSIS. Eimeria dispersa (turkey strain) and Eimeria gallopavonis sporozoites were inoculated into primary cultures of chicken kidney (CK) and turkey kidney (TK) cells. Eimeria dispersa sporozoites were more infective in either cell type than those of E. gallopavonis: at 4 hr, the percentage of infection was 67-98 for E. dispersa but only 23-56 for E. gallopavonis. E. dispersa also survived better in culture: at 2 days, losses of E. dispersa in both cell types were only 4-19%, whereas losses of E. gallopavonis were 35-47% in TK cells and 60–95% in CK cells. However, E. gallopavonis developed further than E. dispersa. Location and increase in numbers of intracellular stages at 4 days indicated that E. dispersa proceeded through 2 schizogonic generations before development stopped.

Notes: Book reviewed in this article:Cuatrecasas, P. & Greaves, M. F., eds. 1976, Receptors and Recognition, Vol. I, Series A.

Notes: SYNOPSIS. A mating type analysis was performed on 231 isolates of the cellular slime mold, Polysphondylium pallidum found in 61 samples collected in eastern North America between northern Florida and southern Canada. Seventy-eight percent of the isolates belonged to one of 2 mating types; 18% were incapable of mating with any partner; 3% were homothallic; and 1%, consisting of 2 isolates from a Florida sample, belonged to a separate breeding group. It is suggested that the majority of isolates represent a species capable of local genetic adaptation to a niche, the parameters of which undergo considerable variation over space and time.

Notes: Book reviewed in this article:Avakyan, A. A. 1976. Atlas Anatomii Prosteishikh, Patogen-nykh dlya Cheloveka i Zhivotnykh.

Notes: SYNOPSIS. The ultrastructure of interphase and mitotic nuclei of the epimastigote form of Trypanosoma cyclops Weinman is described. In the interphase nucleus the nucleolus is located centrally while at the periphery of the nucleus condensed chromatin is in contact with the nuclear envelope. The nucleolus fragments at the onset of mitosis, but granular material of presumptive nucleolar origin is often recognizable in the mitotic nucleus. Peripheral chromatin is in contact with the nuclear envelope throughout mitosis, and it seems reasonable to assume that the nuclear envelope is involved in its segregation to the daughter nuclei. Spindle microtubules extend between the poles of the dividing nucleus and terminate close to the nuclear envelope. The basal body and kinetoplast divide before the onset of mitosis and do not appear to have any morphologic involvement in that process. Spindle pole bodies, kinetochores, and chromosomal microtubules have not been observed.

Notes: SYNOPSIS. Developing 2nd- and 3rd-generation schizonts of Eimeria tenella were found in the ceca of chicks infected orally with sporulated oocysts. Several free 2nd-generation schizonts, which varied in diameter from 11 to 21.6 μm, were found on the epithelial surface of the cecum. Some schizonts appeared to have lost merozoites. Other schizonts were intact, one of which was surrounded by an unbroken membrane that followed the contours of the merozoites. Third-generation schizonts, much smaller than 2nd-generation schizonts and with fewer merozoites, were found only on cut or fractured surfaces of the cecal tissue. Third-generation merozoites appeared shorter and thicker than those of the 2nd-generation and were attached to the schizont residuum. A form with conical protuberances and another with 4 triangular segments were found; they were believed to be developing stages 3rd-generation schizonts.

Notes: SYNOPSIS. Endosymbiotic algae from Paramecium bursaria when added to the culture medium are ingested by Chlorella-bearing P. bursaria at a rate of 2,000 algae/organism/day. That the ingested algae are digested and assimilated by the ciliates is suggested by the more rapid growth of Paramecium when algae are added to the medium (G= 40 hr with algae compared to 190 hr without). The digestion by the ciliates of exogenous algae contrasts with the survival of these algae under normal growth conditions. It is suggested that the protection of the endogenous algae is related to their location in peripheral perialgal vacuoles.

Notes: SYNOPSIS. Cell size, macromolecular composition, carbohydrate utilization patterns, and O2 concentrations were measured throughout the growth stages of Naegleria gruberi in agitated cultures in a complex medium. Biphasic logarithmic growth occurred during the initial 83 hr of growth and the mean generation time was 7.0 hr and 19 hr during initial and secondary log growth stages, respectively. The maximum yield was 5 × 10* amebaeJml. The pH rose rapidly (1 pH unit) during the secondary log growth phase (52-83 hr) and continued into the stationary growth phase (83-120 hr). Dry weight, total protein, carbohydrate, and RNA per ameba increased just before the secondary log growth phase. RNA increased 31% to 35% per ameba at the end of each phase of log growth. DNA increased ∼ 2-fold throughout the different growth phases. Average cell size increased 90% during biphasic log growth then decreased during stationary phase. O2 tension decreased from 100% to 18% of saturation during the biphasic growth phase, then increased during stationary growth to near 100% saturation. Glucose and total carbohydrate assays showed little utilization of those substrates throughout the growth stages. Naegleria gruberi presumably has a predominantly aerobic metabolism, also its metabolism may change during the different growth phases.

Notes: SYNOPSIS. Toxoplasma ranae sp. n. is described from the brain of a leopard frog, Rana pipiens, probably from Mexico. Its pseudocysts were 72(55-106) × 48(29-70) μm in fixed sections. They contained an average of ∼ 4,000 slightly curved elongate zoites measuring 4–5 × 0.5 μm, with a central, spherical, vesicular nucleus.

Notes: Book reviewed in this article:Pennycuick, C. J. 1974. Handy Matrices of Unit Conversion Factors for Biology and Mechanics.

Notes: SYNOPSIS. Egestion of carmine particle-containing food vacuoles from the cytoproct of Tetrahymena pyriformis has been analyzed by high-speed cinemicrography. The vacuole may enter into position in the cytoproct ∼ 7 sec before ejection, and forms a distinct bulge beyond the outline of the cell surface for over 2 sec prior to ejection. The ejection process itself requires 20–80 msec.

Notes: SYNOPSIS. Blood films were examined from 1477 birds of Taiwan (193 species, 49 families). Haemoproteus Kruse was by far the commonest parasite, with Leucocytozoon Danilewski a not very close second. It is probable that some of the Haemoproteus infections represented new species, and 1 occurring in the Bamboo Partridge (Bambusicola thoracica sonorivox Gould) seemed characteristic enough to justify recognition as such; the name Haemoproteus bambusicolae sp. n. is proposed for this organism. Malaria was found in 77 birds, the greatest number of infections occurring in the Bamboo Partridge. Most of them were caused by Plasmodium juxtanucleare Versiani & Gomes, a pathogen of chickens, but a number were due to an undetermined species of Plasmodium. The Bamboo Partridge may be a reservoir host of the former. A few other identified species (P. rouxi Sergent & Sergent, P. hexamerium Huff, P. tenue Laveran & Mesnil) were seen, as well as some unidentified ones. Plasmodium tenue was seen in Garrulax canorus taewanus Swinhoe, a babbler: until now it was known only from the Pekin Robin (Leiothrix luteus Scopoli), also a babbler, in which we have found it extremely common. Sixty-four microfilarial infections were identified; they were especially frequent in the Button Quail (Turnix suscitator rostrata Swinhoe).

Notes: SYNOPSIS. Short pulse-labeling of log phase Crithidia fasciculata cells with [3H]thymidine allowed the autoradiographic visualization of 2 sites of replication of kinetoplast DNA situated at the periphery of the networks and separated by 180°. Longer pulse-labeling led to the previously reported total peripheral labeling pattern. Pulse-labeled networks possess an intermediate density in ethidium bromide-CsCl equilibrium gradients between the densities characteristic of closed networks and open or linear DNA. Removal of ethidium bromide by several methods and treatment of intermediate band networks with RNase and pronase had no effect on the equilibrium rebanding pattern. Closed minicircles of Leishmania tarentolae are not labeled by a short pulse of intact cells with [3H]thymidine. A chase of ∼ 3–4 hr is required for the appearance of radioactivity in closed minicircles, a time delay which implies the existence of intermediate events between replication and eventual covalent closure of the minicircles.

Notes: SYNOPSIS. Epizootic outbreaks of red-sore disease in several reservoirs in the southeastern United States have been reported to cause heavy mortality among several species of fish having sport and commercial value. The etiologic agent is said to be the peritrich ciliate Epistylis sp.; secondary infection by the gram-negative bacterium Aeromonas hydrophila produces hemorrhagic septicemia which results in death. However, in recent studies on the largemouth bass Micropterus salmoides, Epistylis sp. could be isolated from only 35% of 114 lesions from 114 fish, while A. hydrophila was found in 96% of the same lesions. Transmission and scanning electron microscopy of lesions associated with red-sore disease indicate that neither the stalk nor the attachment structure of Epistylis sp. have organelles capable of producing lytic enzymes. Since other investigators have shown that A. hydrophila produces strong lytic toxins, and in absence of evidence to the contrary, it is concluded that Epistylis sp. is a benign ectocommensal and that A. hydrophila is the primary etiologic agent of red-sore disease.

Notes: Weiser, Jaroslav. 1977. An Atlas of Insect Diseases, 2nd ed. Dr. W. Junk, B. V., Publishers, P. O. Box 3713, The Hague, Holland and Academia Publishing House, Czechoslovak Academy of Sciences, Prague, Czechoslovakia. 81 + 240 pp. 75 Dutch Guilders.

Notes: SYNOPSIS. The structure and cytochemistry of spores of Myxobolus sp. from plasmodia which occur in the gill filaments of the common shiner Notropis cornutus were studied by light microscopy and by scanning and transmission electron microscopy. The thin-walled valves of the pyriform spores are thickened in the lateral sutural and apical regions. Mucous material is associated predominantly with the posterior end of many spores. The plasmodium is surrounded by a syncytial wall bounded by 2 membranes. Pinocytotic channels are formed by the inner membrane and numerous dense vesicles are pinched off at the distal ends of the channels. Sporogenesis is initiated by the envelopment of one vegetative cell by another. The larger, enveloped cell divides to form a disporous pansporoblast, which contains 2 pairs of capsulogenic and valvogenic cells and 2 binucleate sporoplasm cells. Each capsular primordium and connecting external tubule gives rise to a polar capsule which houses a helically coiled polar tubule. The apical end of each polar capsule is plugged by a stopper. The valvogenic cells surround the capsulogenic and posteriorly situated sporoplasm cells to form the spore valves. Iodinophilic (glycogen) inclusions were not seen in spores stained with iodine or Best's carmine. A darkly stained band was observed around the posterior region of most spores stained with Best's carmine. In the electron microscope large aggregates of β glycogen particles were seen in the cytoplasm of sporoplasm cells in mature spores.

Notes: SYNOPSIS. DNA synthesis during growth and differentiation in Naegleria gruberi strain NEG populations has been studied. Autoradiography of cells labeled with [3H]thymidine revealed that grains are concentrated over the nuclei in logarithmically growing populations of cells, whereas in differentiating cells, grains are scattered over the cytoplasm; i.e. no significant nuclear labeling is detectable. It was established by MAK chromatographic analysis that [3H]thymidine is incorporated into double-stranded DNA in Naegleria and that the actual amount of incorporation in the logarithmically growing populations of cells is 20 times greater than that in differentiating cells. These results suggest that nuclear DNA synthesis is reduced markedly soon after the initiation of differentiation, while cytoplasmic DNA synthesis continues. It was established from cell cycle analysis that the approximate intervals of G1, S, G2, and M phases were 180, 183, 90, and 28 min, respectively. Hence, the reduction in the nuclear DNA synthesis in differentiating cells is not due to the inhibition of initiation of DNA replication, but rather to the termination of the DNA replicating process. Thus DNA synthesis is curtailed in the presence of RNA and protein synthesis which are required for differentiation.

Notes: SYNOPSIS. The activity and sedimentation of acid phosphatase (APase), acid deoxyribonuclease (DNase), and acid ribonuclease (RNase) were investigated throughout growth and encystment in Acanthamoeba castellanii. The activities/mg protein of all 3 hydrolases are high in young cultures and decrease to constant levels in postlog cells. The RNase activity/ ameba decreases 50% during growth, whereas the activity/cell of both APase and DNase remains constant. The percent sedimentation at 20,000 g of all 3 enzymes gradually increases from about 40% in midlog to a plateau of 80% in postlog cells. During encystment, the sedimentation behavior of RNase differs from that of APase and DNase. Encystment is characterized by a differential decrease in the activity/cell of the 3 hydrolases, with RNase decreasing most rapidly and APase least rapidly. APase is unique in that a transient increase of its specific activity is noted during encystment, even though its activity/cell is decreasing.

Notes: SYNOPSIS. Eighteen strains of algae, including 17 exsymbiotic from Paramecium bursaria, were tested for infectivity for P. bursaria, syngen 2 aposymbiotes, and Concanavalin A (Con A) agglutinability. All 6 infective algal strains were relatively resistant to agglutination by Con A, suggesting that algal surface characteristics are correlated with infectivity. Among the noninfective strains, high and low agglutinability were about equally represented, indicating that the Con A titer alone is not a sufficient indicator of infectivity. It is suggested that noninfective algal strains are the progeny of mutations occurring within the endozoic population and fortuitously selected by the external culture medium.

Notes: SYNOPSIS. Studies on sexual compatibility among populations of different origins of axenically cultivated, heterotrophic, homothallic dinoflagellates resembling Crypthecodinium cohnii have been undertaken using motility mutants and complementation testing. We showed previously that one interbreeding group or biologic species can be found on the East and West coasts of the United States, thus far associated only with Fucus. In addition, we found several strains of these organisms each of which, though fertile in intrastrain crosses, appeared infertile with all other isolates. A continuation of these studies has revealed another biologic species of wide geographic distribution (Taiwan, California, Florida, New York) and apparently associated with sea weeds other than Fucus.

Notes: SYNOPSIS. The effect of pretreatment with Isospora felis and bacillus Calmette-Guérin (BCG) on the reexcretion of Toxoplasma gondii occysts was studied in 16 coccidia-free cats. The following conclusions were drawn: (A) Chronically T. gondii-infected cats reexcreted T. gondii oocysts after superinfection with I. felis, and this reexcretion was prevented in cats infected with I. felis before T. gondii infection. (B) Administration of BCG before Toxoplasma infection had no apparent effect on the outcome of the infection.

Notes: SYNOPSIS. In cultures of Platymonas subcordiformis Hazen, grown in appropriate light-dark cycles, as many as 75% of the cells adhered to the surface of the glass culture vessel toward the end of the light period of each day. Cell division occurred primarily while the cells were attached. Subsequently, motile daughter cells were released into the growth medium by the rupture of the mother cell theca. The settling behavior appears to be an integral part of the life cycle being synchronized to the same extent as cell division.

Notes: SYNOPSIS. The coccidian parasite Eimeria gadi was found in the haddock, Melanogrammus aeglefinus, taken from the Nova Scotian fishing banks. The haddock infection rates ranged from a high of 58% on Emerald Bank to a low of 4% on Georges Bank, the average being 32%. There was no relationship between sex and degree or prevalence of infection. Although the probability of an occurrence of infection increased with size, small fish with heavy infections were observed. The degree of infection had no apparent effect on the condition factor (length/weight) of the fish. The infection rate reached a maximum in the fall of the year while the heaviest infections were observed in the spring. It is evident from the data that the infection is fatal.The parasite mass, appearing as a creamy viscous to a yellow semisolid material in the swimbladder, consisted of various parasite stages, fibrous and cellular debris, and lipid material. Some aspects of the sporocyst stage are described.No other gadoids from the Nova Scotian banks were found to be infected; however, a single specimen of the fourbeard rockling, Enchelyopus cimbrius, from St. John's, Newfoundland, was found to be heavily infected with E. gadi.

Notes: SYNOPSIS. Synthesis of RNA in the macronucleus and appearance of RNA in the cytoplasm were studied in heat synchronized Tetrahymena pyriformis GL and compared to those found under conditions of logarithmic growth (28 C) and during heat shocks (34 C).In macronuclei of logarithmically growing cells precursors were processed to 2 rRNA species (25S and 17S). In addition, another RNA (15S), more homogeneous than the RNA (8-15S) in the cytoplasm, was observed in the macronucleus. Both 17S and 25S rRNA species were found in the cytoplasm, 17S rRNA appearing more rapidly than 25S rRNA. Synthesis of rRNA was suppressed at 34 C in cells subjected to heat synchronization; 8-15S RNA synthesis appeared to be inhibited to a lesser extent.During the time preceding the first synchronized division, the synthesis of rRNAs in the macronucleus slowly recovered. Early in the cycle, almost no newly synthesized rRNAs were extracted. By 30 min after the last heat shock (EH), most of the RNA synthesized was not identified as rRNA. By 60 min after EH, the pattern of RNA synthesis had not returned to that observed in logarithmically growing cells.

Notes: SYNOPSIS. Cell-free preparations of Acanthamoeba castellanii trophozoites transfer glucose from UDP-[U-14C]glucose to a chloroform-soluble form. This radioactive material has been isolated by thin-layer chromatography; it contains an alkali-labile and an alkali-stable (unsaponifiable) component. Treatment of the enzymic product with 0.1 N KOH for 15 min at 0 C or 20 C releases radioactivity into the aqueous phase as glucose. During this treatment, 30–60% of the original glycolipid remains chloroform-soluble. It is considered to be an alkali-stable glycolipid because no further loss of radioactivity occurs during an additional 45-min of treatment with 0.1 N KOH. During incubation with 0.1 N HCI at 100 C glucose is released quantitatively from both the untreated glycolipid and the alkali-stable glycolipid with a half-time of 6 min. Glycolipid formation is inhibited by UDP and is reversible; extracts catalyze the formation of UDP-glucose from the alkali-stable glucolipid and UDP.The chemical and physical properties of the alkali-stable glycolipid are consistent with a glucosyl phosphoryl polyprenol structure. Extracts prepared from cysts catalyze the formation of glycolipids aiso, but the glucosyltransferase activity/cell decreases during the course of encystment. Radioactivity is incorporated into the fraction insoluble in chloroform-methanol-water (1:1:1:) during these incubations when UDP-[U-14C]glucose or [14C]glycolipid is the substrate.

Notes: SYNOPSIS. In populations of Euglena gracilis strain Z synchronized by cultivation on a repetitive light-dark cycle, chloroplasts undergo cyclic changes in structure. During most of the light period chloroplasts are relatively compact with closely appressed lamellae; during the dark (division) period the chloroplasts become quite distended. This change persists for at least one cycle even when the cells are left in continuous light, suggesting that the periodicity may be related more to the age of the cell than to a direct effect of light. In addition, the pyrenoid in synchronized cells has a transient existence, being present only in the first half of the light period.

Notes: SYNOPSIS. Although large hemoglobin inclusions are observed in intraerythrocytic Babesia microti parasites, they are absent from parasites freed of hamster red cells by immune lysis with antihamster erythsocyte serum. Babesia microti has no cytostome. This parasite, therefore, does not appear to feed by phagocytosis of large boluses of hemoglobin, as does Plasmodium. To determine whether Babesia can pinocytose protein, free parasites were fed ferritin in an in vitro system. Ferritin was taken up from the entire cell surface into narrow channels within 15 min at 37 C. Only merozoites, with their pellicular complex, failed to take up the protein. By 60 min, the ferritin was highly concentrated in many channels and vesicles, which formed interconnecting stacks. The ferritin-containing channels became associated with membrane whorls of the multimembranous structure. Membrane whorls were also observed in the process of extrusion in samples incubated for longer times. These events may represent steps in the digestion and excretion of the pinocytosed protein. Empty channels formed when Babesia was fed albumin. The diaminobenzidine reaction for hemoprotein was positive for the channels in both free and intraerythrocytic babesias. The staining reaction was completely inhibited by cyanide, but not at all by aminotriazole. These results further suggest that Babesia pinocytoses hemoglobin in vivo. Plasmodium lophurae parasites freed of red cells by immune lysis are surrounded by 2 membranes and apparently can ingest ferritin only through the cytostome. Extracellular cytostomal feeding involves both membranes, as it does in vivo. Ferritin was found in food vacuoles, some of which contained hemoglobin ingested before parasite isolation, connected to or near the cytostome. In both Plasmodium and Babesia low temperature inhibited ferritin uptake.

Notes: SYNOPSIS. During the course of infection in the rat, Trypanosoma lewisi produces 2 antigenic variants: the 1st represents the initial, reproducing population of cells; and the 2nd the nonreproducing, ablastin-inhibited adult population. The specificities of the agglutinins elicited by the variants were studied by adsorption and agglutination methods and the newer immunoelectroadsorption technic. It was found that the reproducing variant has a surface antigen that reacts with the agglutinin specific for the adult variant, but this antigen does not become immunogenic until transformation to the adult variant occurs. It was also found, with fractions of immune sera obtained by gel filtration, that the agglutinin specific for the reproducing variant is IgG and that specific for the adult variant, IgM. The antigenic variants of pathogenic and nonpathogenic trypanosomes are compared, and the roles of trypanocidal and ablastic antibodies in the induction of antigenic variation are discussed.

Notes: SYNOPSIS. Living Trypanosoma musculi bloodstream trypomastigotes were agglutinated specifically with concanavalin A (ConA), wheat germ agglutinin (WGA), soybean agglutinin (SBA), and fucose-binding protein (FBP). The agglutination with these lectins of living cells from which the coat was removed by trypsinization was the same as with intact trypanosomes. Glutaraldehyde or formalin fixation did not affect the results with regard to agglutination with WGA, SBA, and FBP, but lower agglutination with ConA was observed upon fixation. By using a dense iron-dextran marker many fewer ConA marker particles were localized at the fine structural level in the intact than in trypsin-treated trypanosomes. On the basis of the results obtained by agglutination and electron microscopy, it is likely that fixation cross-links intact surface-coat components associated with the ConA binding sites. It is evident from the studies in which lectins were employed that ligands containing α-D-mannose, N-acetylglucosamine, N-acetylgalactosamine, and α-L-fucose are randomly distributed in the outer surface of the pellicular and flagellar membranes of T. musculi trypomastigotes. Results obtained with α-amylase- and dextranase-treated trypanosomes suggested that lectin-binding sugar ligands in the cell surface were not directly associated with α-1,4 or repetitive α-1,6 glucan-bonded polysaccharide moieties. Similar conclusions can be drawn on the basis of neuraminidase treatment with regard to N-acetylated neuraminic acids.After thorough washing, intact, but not trypsin-treated trypomastigotes were agglutinated specifically with antisera against whole mouse serum and against mouse IgG. Evidently, adsorbed constituents of mouse serum are regular components of the T. musculi surface coat. After incubation in dilute whole mouse serum or in mouse IgG solutions, also the trypsinized cells were agglutinated by the 2 antisera. No such results were obtained with trypsinized cells incubated in serum-free buffers. It was concluded that mouse serum proteins were readily readsorbed on, and firmly bound to the trypsinized cells' surfaces. Specific agglutinations were obtained with trypsinized cells after incubation in dilute rat, rabbit, bovine, and human sera and in solutions of rat and rabbit IgG in reactions with the corresponding antisera. It seems, therefore, that the host serum proteins are adsorbed nonspecifically to the cell surface of trypsinized T. musculi bloodstream forms.When examined by electron microscopy, the intact trypomastigotes were covered by an ununiform, slightly granular, fibrillar extracellular coat, applied to the entire outer lamina of the pellicular and flagellar membranes. No indication of such a coat was noted in the trypsinized organisms. Flocculent surface coat-like matrix could, however, be discerned in cells which, after trypsinization, were incubated in various sera.

Notes: SYNOPSIS. Leptomonas pessoai from the reduviid hemipteron Zelus leucogrammus, was cloned. The original strain (ATCC 30252) and clones can differentiate from promastigote to opisthomastigote. Differentiation is faster at high temperature (37 C) and in a defined medium as compared with a complex medium. It is suggested that Leptomonas pessoai be designated Herpetomonas samuelpessoai.

Notes: Book reviewed in this article: Journal of Toxicology and Environmental Health. Vol. 1, no. 1, Sept. 1975. Hemisphere Publishing Corp.Maloine, S. A., ed. 1974. Diagnostique Immunologique des Parasitoses a Protozoaires et Helminthes.

Notes: SYNOPSIS. Haemogregarina bigemina was found in all Blennius pholis which exceeded 5.0 cm in length, but in none measuring less than 3.5 cm. No exoerythrocytic development was recorded. The first B. pholis eggs hatched in May while the first patent infections of H. bigemina occurred from September onward in metamorphosed fish. Consequently, if the life cycle of H. bigemina includes a vector, that organism is active between May and September at least. Circumstantial evidence indicates that the hematophagous isopod, Gnathia maxillaris and not leeches, could be a vector of H. bigemina. Developmental stages of sporozoa were found in a small number of the isopods which had fed on infected B. pholis but the parasites could not be identified as H. bigemina with certainty. Subcellular organization, typical of sporozoa, was recorded by electron microscopy of H. bigemina.

Notes: SYNOPSIS. Leptomonas pessoai promastigotes cultivated in a synthetic medium were agglutinated with concanavalin A (Con A). Agglutination was predominantly of the flagellar-flagellar type, and was inhibited with sucrose or α-methyl-D-mannoside.Cell surface polysaccharides and/or glycoproteins were demonstrated by several cytochemical methods at the fine-structural level. A Con A-horseradish peroxidase-diaminobenzidine (DAB) technic was used to detect Con A receptors in the pellicular membrane of the flagellar pocket region and in the flagellar membrane proper. Somewhat less of the Con A-DAB reaction product was observed in the pellicular membrane enclosing the rest of the cell.

Notes: SYNOPSIS. Phylogenetic relations are ultimately determined by homologous relationships, including both structural and functional data. Following the establishment of those relationships, the direction of evolutionary change must be determined using paleontologic, developmental, and especially morphocline, data. From that perspective the direction of subsequent development becomes clearer and the problems of origins become more explicit.Using the foregoing methodology it has been possible to identify plausibly monophyletic groups of animal protists or protozoa. Allowing for attendant difficulties, there nevertheless emerges certain fairly convincing generalities: (a) the predominantly pseudopodial forms, with a few minor exceptions, have direct origins from apochlorotic algae; (b) the predominantly kinetidal forms (zooflagellates and ciliates), though also derived originally from apochlorotic algae, give evidence of extended evolutionary development with the especially noteworthy emergence of a permanent ingestatory structure; (c) both groups have increased size, a tendency towards multinuclearity and polyploidy, cytoplasmic differentiations of various sorts, and complex life cycles.In terms of further evolution, namely the emergence of multicellular animals, the pseudopodial forms are almost certainly a dead end while the kinetidal forms are arguably the ancestors of at least 2 metazoan phyla.

Notes: SYNOPSIS. A clone of Cyclidium citrullus, adapted to growth at 18, 27, 37, 43, and 46 C had an optimum at 43 C, with 6.5 divisions/day. Transfer of cells previously grown at 43 or 46 C to 18 C resulted in death of most of the cells, transfer to 27 C increased the lag period, and transfer from 18 C to 37 or 46 C was followed faster division. All cells died at 48 C; some divided before death. At the temperatures employed maximum cell sizes (length and width) were achieved in the early log phase. At 43 C, however, the early log phase cells were smaller. Quantitative and qualitative differences in the free amino acids in the cells were found in ciliates grown at 18, 43, and 46 C; the highest amount/cell was found at 18 C, and the lowest at 43 C. High concentration of proline was noted only at 18 C.

Notes: SYNOPSIS. Additional information on host interactions with trypanosomatid membranes was obtained from studies of a monomorphic strain of Trypanosoma brucei harvested at peak parasitemia from intact and lethally irradiated rats. Pellets of trypanosomes were fixed briefly in glutaraldehyde and processed for thin section electron microscopy or freeze-cleave replicas. Observations of sectioned material facilitated orientation and comparison of details seen in replicas. Fracture faces of cell body and flagellar membranes as well as 3-dimensional views of the nuclear membrane were studied. Cell body membranes of 80% of the organisms from intact rats contained random arrays of intramembranous particles (IMP). Aggregated clusters of particles appeared on the fracture faces of 20% of the trypanosomes. Some of these membranes had nonrandomly distributed particles aligned in distinct rows on the outer fracture face of both cell body and flagellum. Many inner face fractures of the cell body membranes had a particle arrangement similar to the longitudinal alignment of cytoskeletal microtubules. No aggregated particle distribution was seen in membranes of trypanosomes harvested from lethally irradiated rats. Replicas of trypanosome pellets also had plasmanemes as a series of attached, empty, coated membrane vesicles. These structures were found in close association with, as well as widely separated from the parasites. The shedding of these vesicles and the variation of particles in cell body membranes are discussed in light of antibody-induced architectural and antigenic changes in surface properties of trypanosomatids.The convex face of the inner membrane of the nucleus also is covered with randomly arrayed particles. More IMP were observed on the inner than on the outer nuclear membranes. Images of nuclear pores were also seen. The importance of these structures in drug and developmental studies of trypanosomes is discussed.On fracture faces of the flagellar membrane there were miniature maculae adherentes, unique to the inner fracture face and occurring only at regions of membrane apposition between cell body and flagellum. Each cluster of particles exposed by the freeze-cleave method corresponds to an electron-dense plaque seen in thin section images. However, because of a unique fracture pattern, these plaques were not revealed on the apposing body membranes, as illustrated in thin sectioned organisms.

Notes: SYNOPSIS. Phytomonas davidi (Trypanosomatidae) originally discovered by Lafont in 1909 on the island of Mauritius was rediscovered in Euphorbia cyathophora in Florida. Successful cultures were established in diphasic medium consisting of duck blood agar and modified Phillips’medium as overlay. Optimal growth was obtained when Mansour's medium was used as overlay and poorest growth when Cowperthwaite's medium buffered at pH 5.0 was utilized for this purpose. Marked changes tending toward choanomastigotes rather than the elongate twisted promastigotes were observed in cultures.

Notes: SYNOPSIS. Past reports of Synophrya, the only apostome ciliate known to harm its host, indicated that it was restricted in its distribution and in the hosts it can infect. In recent collections of decapods from the off-shore waters of North Carolina and Georgia, 30% of all specimens were infected with Synophrya. Forty-four percent of all the species collected had individuals infected with Synophrya, and 63% of all the families collected contained species that had infected individuals. There is no obvious structural or phylogenetic relationship between these families that explain why they are infected. The presence of Synophrya may be related to salinity since decapods captured in estuaries were never infected and decapods captured close to shore were only rarely infected. The salinities of these waters range from 15–35%, but the salinity of the off-shore waters where the great majority of infected specimens was found ranges only from 35–36%.

Notes: Book reviewed in this article:Pirt, S. J. 1975. Principles of Microbe and Cell Cultivation.

Notes: SYNOPSIS. Plasmodium lophurae serine hydroxymethyltransferase (EC 2.1.2.1) was partially purified and characterized by (NH4)2SO4 fractionation and chromatography on Sephadex G-100. The enzyme, precipitated by 3.0–3.3 m (NH4)2SO4, had a molecular weight of 68,300 as estimated by exclusion chromatography on G-100. The pH optimum of the enzyme was 6.8–7.6 in sodium phosphate-citrate buffer. Citrate stabilized the enzyme during storage in phosphate buffer at 4 C. The Km was 4.3 × 10−3m for l-serine and 2.5 × 10−4m for tetrahydrofolate.

Notes: Book reviewed in this article:Cordero del Campillo, M. et al. 1975. Indice-Catalogo de Zooparasitos Ibéricos. I. Protozoos. II. Trematodos.

Notes: SYNOPSIS. Haemogregarina balli sp. n. is described from the blood and organs of the common snapping turtle Chelydra serpentina serpentina and from the gastric and intestinal ceca of the presumed invertebrate hosts, the leeches Placobdella parasitica and Placobdella ornata. In the peripheral blood of the turtle, male and female gametocytes and immature erythrocytic schizonts are found within erythrocytes. The maturation of erythrocytic schizonts containing 6–8 merozoites is recorded from liver imprints. Schizonts with 13–25 merozoites are found in various cells of the liver, lung and spleen. In the gastric ceca of the leeches the host erythrocytes are digested, releasing the gametocytes and immature erythrocytic schizonts. Immature erythrocytic schizonts degenerate. Association of the gametocytes occurs in the intestinal ceca. The microgametocyte apparently gives rise to 4 nonmotile microgametes, one of which fertilizes the macrogamete while the other remain as condensed, residual nuclei on the periphery of the developing oocyst. The oocyst increases in size with maturity. A mature oocyst produces 8 sporozoites from a single germinal center. Sporozoites liberated from the oocyst are found in the tissues of the leech. Transovarial transmission of the parasite does not occur in the turtle. Attempts at experimental transmission failed. Previously unfed (control) leeches were negative for the parasite. Haemogregarina balli is compared with other haemogregarines described from C. serpentina. Features of species of Haemogregarina and Hepatozoon as well as the taxonomy of these genera are discussed.

Notes: SYNOPSIS. In low viscosity media, Euglena gracilis strain Z responds to a sudden change in light intensity by a cessation of forward movement, followed by a reorientation of the locomotor flagellum which results in turning of the cell around the lateral axis (photophobic response). At a viscosity interface between low [∼ 1 cP (centipoise)] and high (4000 cP) media, the cells exhibit avoidance responses or become immobilized in the higher viscosity medium. Upon changing the light intensity, free swimming cells have photophobic responses, while immobilized ones undergo body contractions. For cells immersed in media of varying viscosity, the delay between light stimulation and body contraction (transduction time) is shortest at high viscosities. From 500 to 2000 cP, where the cells are capable of both movement and light-induced body contractions, there is a logarithmic dependence of the transduction time on the viscosity. The transduction time does not vary appreciably with the intensity of the primary light stimulus within a range of 0.14-1.13 kW/m2.

Notes: SYNOPSIS. Eight species representing 8 genera of gallinaceous birds were used: Alectoris graeca; Colinus virginianus; Coturnix coturnix; Gallus gallus; Meleagris gallopavo; Numidia meleagris; Pavo cristatus; Phasianus colchicus. Three-week-old birds were dosed with sporulated oocysts of Eimeria tenella Beltsville strain. At 4, 24, 48, 72, 96, 120, 144, and 168 hr after inoculation, 1-3 infected birds and uninoculated controls of each species were killed by cardiac exsanguination. Pieces of intestines were fixed and examined for stages of E. tenella as stained paraffin sections or indirect fluorescent antibody preparations. Oocyst counts were made in droppings collected for the first 6 days of the patent period. Sporozoites were found in the lamina propria of some birds of 5 species at 4 hr postinoculation, but no stages were found thereafter except in the breeds of G. gallus and A. graeca. At 144 and 168 hr postinoculation, a few macrogametes were found in the ceca of 2 A. graeca, but no oocysts were found in the feces. No statistical difference was found between the number of oocysts produced/bird in the breeds of G. gallus examined. It is evident from these observations that E. tenella did not complete its life cycle in several close phylogenetic relatives of G. gallus, even though in other studies this parasite was found to complete its life cycle in cell cultures derived from the same birds.

Notes: Book reviewed in this article:Faust, E. C., Beaver, P. C. & Jung, R. C. 1975. Animal Agents and Vectors of Human Disease.

Notes: SYNOPSIS Previous surveys of feral Drosophila melanogaster in Clarke County, Georgia, indicated 10 to 30% of the flies were parasitized with Herpetomonas ampelophilae. In the present study, herpetomoniasis in D. melanogaster reared in population cages rose from 0.01% to 90% in 14 days. The infection fluctuated between 5% and 100% during the 250 days in which the ageledeme was maintained. Herpetomonas were found endotrophically, peritrophically, and in the Malpighian tubules. In population cages, although there were sporadic rises in percentages, peritrophic and Malpighian tube infections usually remained low. Endotrophic infections often reached 100%, with individual flies being heavily parasitized. Although Chatton felt that several species of Herpetomonas inhabit a single species of Drosophila, we suggest that all the Herpetomonas in D. melanogaster be referred to as H. ampelophilae.

Notes: SYNOPSIS Experiments were designed to study intestinal pathophysiologic changes associated with coccidial infections in mammalian hosts. Pairs of male Sprague-Dawley rats were killed at various times postinoculation (PI) with 104 or 106 sporulated occysts of Eimeria nieschulzi. The small intestine from each rat was removed, weighed, measured, and divided into thirds. From the middle 11 cm of each third, one cm was fixed for histologic examination. Mucosa was scraped from the remaining 10 cm and was assayed for protein content and for peroxidase, sucrase and trehalase activities. Infection with E. nieschulzi was associated with increased mass of the small bowel. Histologically, crypt depth throughout the small bowel was significantly greater (P≤ 0.005) in infected rats than in non-infected ones on PI days 8 and 16. Villus height did not change drastically during low-dose infections (104 oocysts) and varied during high-dose infections (106 oocysts). As a result of these morphologic changes in the mucosa, crypt/villus ratios were usually significantly greater (P≤ 0.005) in all infected rats throughout the small bowel. In general, increased gut weight and changes in crypt and villus dimensions became evident by PI day 2, were most pronounced at PI day 8, and began to return to control values by PI day 16. Peroxidase, sucrase, and trehalase levels equaled or were slightly higher than in controls on PI day 2, dropped significantly below controls (P≤ 0.05) by PI day 8, and returned to, or exceeded control levels by PI day 16. The intensity of all changes was directly dose-dependent.

Notes: SYNOPSIS Random examination of domestic animals revealed the frequent presence of free-living amebae in their bodies. In diseased or dead cows, dogs, pigs, rabbits, pigeons, and turkeys 15 strains of amebae were found, belonging to the genera Acanthamoeba (A. polyphaga), Hartmannella (H. vermiformis), and Vahlkampfia (V. avara, V. enterica, V. inornata)., They were usually accompanied by other infectious agents in different parts of the host bodies. Pathogenicity of 3 isolates could not be demonstrated by inoculation of laboratory animals.Some features of the isolates differed from those previously known for members of these genera. These strains may be considered amphizoic amebae according to Page (1974).

Notes: SYNOPSIS In a hologamy-like process observed in Oxytricha hymenostoma, 2 cells belonging to complementary mating types fused and appeared to contribute equally to the macronuclei of the organism resulting from the fusion. The process was characterized by the absence of micronuclear division and by fusion of the macronuclei of the 2 partners. True conjugation involving micronuclear meiosis was not observed in O. hymenostoma. The biologic significance and the possible adaptive role of the hologamy-like process are discussed.

Notes: SYNOPSIS The mesenteric microvasculature was studied in rats and rabbits infected with Trypanosoma congolense. By examining vessels in the living animals, trypanosomes were observed to adhere to vessel walls by their anterior ends. It was evident from stained preparations of the vessels that the microcirculation contained 4–1400 times as many trypanosomes as were free in the cardiac blood. Parasites were more numerous in very small vessels than in larger vessels, and they were clustered in groups within the small vessels. The localization of T. congolense in the microvasculature is demonstrated and it is shown that this localization is established by attachment of the organism to the vessel wall.

Notes: SYNOPSIS The quantitative direct fluorescent antibody (QDFA) methods were employed for the antigenic analysis of bloodstream forms and culture procyclics of 2 variants, TRUM (Trypanosome Research University of Massachusetts) 106 and TRUM 107, of Trypanosoma brucei brucei. Intact and trypsinized trypanosomes were studied. It was demonstrated that: (A) The specific variant antigens are localized in the surface coat of bloodstream trypomastigotes. (B) In addition to the common antigens shared by bloodstream forms and culture procyclics, there are also certain antigens unique to these latter stages. (C) Still another group of antigens. not found in the culture procyclics, appears to be shared by the bloodstream forms, irrespective of their variant-specific antigens. These antigens may be present in part in the coat or on the cell membrane and in part within the cytoplasm. (D) Irrespective of the bloodstream-form variant from which they are derived, the procyclics are antigenically the same. The QDFA results are analyzed statistically and discussed in the light of the available literature.

Notes: SYNOPSIS Chemical analysis of axenic cultures of Hymenomonas sp. reveals that the degree of coccolith calcification is inversely proportional to the concentration of available nitrogen sources in the medium. This relationship makes possible a useful degree of control over the organism's calcifying activities. In addition, a possible selective advantage of such a relationship in nature is suggested.

Notes: SYNOPSIS. Fine structure and development of Encephalitozoon cuniculi from rabbits were studied in rabbit choroid plexus (CP) cell cultures and were compared to hamster and mouse microsporida.Sporoplasms had a single limiting membrane and contained a large nucleus. Proliferative forms (schizonts) had double outer membranes, the outermost being associated with the formation of the limiting membrane of vacuoles formed within the host cell cytoplasm. These organisms were often binucleate and divided to form sporonts. Sporonts divided once to form 2 sporoblasts which developed into electron-dense spores. Spores had a thick, 3-layered wall and contained a polar filament.The developmental cycle of E. cuniculi in rabbit CP cultures progressed rapidly. Sporoplasms were observed in host cells at 3 hr postinoculation (PI). By 24 hr PI proliferative forms were associated with host cell cytoplasmic vacuoles which contained developing organisms. Mature spores were present in vacuoles by 2 days PI, indicating that the life cycle in the CP system is ∼ 48 hr.The fine structure and the sequential developmental cycle of the mouse and hamster isolates were observed to be identical to those of the rabbit isolate and different from those of the genus Nosema. It is proposed, therefore, that the 3 organisms represent the same species, Encephalitozoon cuniculi.

Notes: RESUME. Deux espèces d'Enteromonas sont observées, provenant, l'une de l'intestin de Triton, l'autre des crottes du Lapin domestique. La cellule piriforme porte un noyau antérieur et 4 flagelles insérts près du pôle ventral du noyau. Le flagelle récurrent (R) est logé dans une dépression ventrale ou cytostome. Les cinétosomes, disposés en une paire antérieure (#1, #2) et une paire postérieure (#3, R), sont liés entre eux par des microfibrilles. Une fibre microtubulaire située au-dessus du noyau est reliée au cinétosome #1. Une autre fibre microtubulaire sous-nucléaire est homologue de la fibre microtubulaire croisée qui existe chez les cellules de Diplozoaires. Le cytostome est bordé par 2 lèvres: la gauche proéminente et armée par plusieurs rangées de microtubules, la droite contenant seulement une mince fibre microtubulaire associée à des microfibrilles. Le cytostome occupe les 2/3 de la face ventrale. Le flagelle récurrent pénètre dans le cytostome puis dépasse l'extrémite de la cellule. Les Bactéries sont phagocytées au fond du cytostome, entre les 2 lèvres distendues. Elles sont digérées dans les nombreuses vacuoles et les corps résiduels sont évacués par rupture de la membrane cellulaire. L'ergastoplasme est concentré près de la périphérie de la cellule. Il n'y a pas de mitochondrie ni d'appareil de Golgi. Dans les kystes observés la cellule plurinucléée est enfermée dans une enveloppe kystique microfibrillaire, les axonèmes sont libres dans le cytoplasme.Les formes diplomonades sont nombreuses et ressemblent aux cellules d'Hexamita, excepté par le cytostome qui est différent. Dans ces formes, les 2 monades sont souvent disposées selon une symétrie axiale binaire mais quelquefois elles sont associées de façon plus anarchique. La cinétide d'Enteromonas est organisée comme celle d'un zoïde de Diplozoaire. Il est possible que le genre Enteromonas soit à l'origine des Diplomonadida et que l'état diplomonadien transitoire chez Enteromonas se soit stabilisé ensuite chez les Diplomonadida. Enteromonas apparaît plus primitif que les autres genres de Diplomonadida aussi nous proposons de créer 2 sous-ordres: celui des Enteromonadina avec le genre Enteromonas et celui des Diplomonadina avec les genres Trepomonas, Trigonomonas, Hexamita, Spironucleus, Octomitus, Giardia. La disposition des cinétosomes et l'existence du cytostome sont les principaux caractères communs entre Enteromonas et les Retortamonadida, cependant les fibres annexes ne sont pas homologues. Une étude plus complète de la division nucléaire et cellulaire de ces 2 ordres de Zooflagellés est nécessaire pour donner un meilleur schéma évolutif.SYNOPSIS. Fine structure of 2 species of Enteromonas, one from the intestine of the salamander, Triturus vulgaris, and another from the feces of domestic rabbit, Oryctolagus cuniculi, is described. The pyriform cell has an anteriorly located nucleus. The 4 flagella originate from an area near the anterior end of the nucleus. The recurrent flagellum (R) is lodged in a ventral depression or cytostome. The kinetosomes, arranged into 2 pairs, anterior (#1, #2) and posterior (#3, R), are interconnected by microfibrils. One microtubular fiber, connected to kinetosome #1, is situated near the anterior surface of the nucleus. Another, subnuclear, microtubular fiber is homologous to the “crossed'’fiber found in Diplozoa. The cytostome is bordered by 2 lips: the preeminent left lip is equipped with several rows of microtubules, while the right lip contains only a thin microtubular fiber associated with microfibrils. The cytostome occupies 2/3 of the ventral surface. The recurrent flagellum passes over the anterior surface of the cell and then comes to lie in the cytostome. The bacteria are phagocytosed in the bottom part of the cytostome between the 2 distended lips. They are digested in numerous vacuoles. The undigested residual bodies are evacuated by a rupture of the cell membrane. The ergastoplasm is concentrated near the cell periphery. Mitochondria and the Golgi apparatus are absent. In the cyst stage, the multinucleate cell is enclosed in a microfibrillar membrane; the axonemes lie free in the cytoplasm.Diplomonad forms of Enteromonas resembling Hexamita are numerous, except that the cytostome is different in these 2 genera. In such forms, the arrangement of the 2 individuals often has binary axial symmetry, but on occasion they are associated in a more anarchic fashion. The mastigont of Enteromonas is organized like that of a single zooid of a diplozoon. It is possible that the genus Enteromonas is ancestral to Diplomonadida and that the diplomonad state, transitory in Enteromonas, became permanently established in Diplomonadida. Enteromonas appears to be more primitive than the other genera of Diplomonadida. Thus we propose 2 suborders: Enteromonadina, subord. nov. with the genus Enteromonas, and Diplomonadina Wenyon, emend., with the genera Trepomonas, Trigonomonas, Hexamita, Spironucleus, Octomitus, Giardia. The arrangement of the kinetosomes and the existence of a cytostome are the principal characters common to Enteromonas and Retortamonadida, while their “accessory'’fibers are not homologous. A more complete study of division of the 2 zooflagellate orders is necessary for the presentation of a more detailed evolutionary scheme of these groups.

Notes: SYNOPSIS Enhanced esterase C activity could be demonstrated by starch gel electrophoresis in various stocks of Paramecium spp. (P. primaurelia stocks 90 and 540, P. biaurelia stock 93, P. tetraurelia stock 29. P. pentaurelia stock 87, P. octaurelia stocks 31 and 300, and P. multimicronucleatum species 3, stock 8 MO) grown in Adaptation Medium. This esterase, however, was barely detectable when they were cultivated in Axenic Medium. Addition of trypticase to Adaptation Medium resulted in reduction of esterase C in the ciliates. This effect is ascribable to Na acetate present in trypticase. Since esterase C increased with the decrease in acetate concentration (as estimated by gas-liquid chromatography) during growth of Paramecium, acetate appears to be utilized by the cells. Sensitivity of esterase C to acetate occurs in all 6 species of Paramecium examined. Different stocks within a species may have different levels of sensitivity; in one case this is genetically determined. The results emphasize the importance of controlling and manipulating growth conditions for the assessment of inter- and intraspecies variations in the isozymes of Paramecium.

Notes: SYNOPSIS Vitamin B12 deficiency arrests cell division in Euglena gracilis. B12 starvation for short periods made it possible to induce synchronous growth by addition of the vitamin. Culture conditions were established to optimize replenishment synchrony. The DNA content of E. gracilis in steady state culture and vitamin B12 deficiency culture was measured by flow cytofluorometry and was consistent with colorimetric determinations. The cell volume and DNA distributions of E. gracilis in synchronous culture were analyzed and the sequential changes during the division cycle were computed. Synchronous culture permits more definitive studies of shifts in cell volume and DNA distributions, in which the biochemical events required for cell division are presumably synchronized.

Notes: Atypical eukaryotic RNA polymerase activity was demonstrated in nuclei of Crypthecodinium cohnii, a eukaryote devoid of histones. Nuclei were isolated from growing cultures of this dinoflagellate and assayed for endogenous RNA polymerase (EC 2.7.7.6) activity. There was a biphasic response to Mg2+ with optima at ˜ 0.01 and 0.02 M MgCl2, but in contrast to other eukaryotic RNA polymerases, this enzyme activity was inhibited by low MnCl2 concentrations. In the presence of 0.01 M MgCl2 the optimum (NH4)2SO4 concentration was 0.025 M, a concentration at which the nuclei were lysed. Incorporation of [3H]UMP into RNA was inhibited by actinomycin D and dependent on the presence of undegraded DNA, and the reaction product was sensitive to ribonuclease and KOH digestion. Omission of one or more ribonucleoside triphosphates greatly reduced the incorporation. Only a slight enhancement of RNA polymerase activity resulted from the addition of various amounts of native and denatured calf thymus DNA. Spermine caused a marked inhibition while spermidine had little effect on RNA synthesis in the nuclei. Under the optimum conditions described in the present paper the nuclei incorporated ˜ 3 pmoles of [3H]UMP/muml; DNA at 25 C for 15 min, and ˜ 80% of this activity was inhibited by the eukaryotic RNA polymerase II inhibitor, α-amanitin (20 m̈/ml). A unique situation therefore exists in C. cohnii nuclei, in which absence of histones (a prokaryotic trait) is combined with α-amanitin-sensitive RNA polymerase activity (a eukaryotic trait).

Notes: After demonstration that emetine is amebicidal by inhibiting protein synthesis, the question arose whether active protein synthesis is required for emetine's amebicidal effect. The answer appears to be “no,” as derived from experiments on intact amebae. Responses were compared for log- and stationary-growth phase amebae. In the latter, protein synthesis is significantly slower, and sensitivity to emetine, i.e. degree of inhibition of protein synthesis, was maintained independently of rate of protein synthesis. Both stages equally bound tritiated emetine to their ribcsomes. Binding of [3H]emetine was not affected by certain drugs that interfere with energy metabolism, protein synthesis, and/or ribosomal function, e.g. dinitrophenol, puromycin, chloroquine, and acriflavin. High concentrations of EDTA combined with puromycin (which disaggregates ribosomes into their subunits) lowered binding by 50%.In chase experiments the ribosomes of intact amebae were prelabeled with [3H]emetine or [3H]isoemetine, then exposed to relatively high concentrations of unlabeled emetine. Labeled isoemetine was displaced almost completely, whereas no displacement of [3H]emetine occurred; evidently, the high stability of the emetine-ribosome binding is due in part to a hydrogen-bonding reaction of the C-1' atom of the emetine molecule with the chain-elongation site. Finally, evidence was obtained that capacity to bind emetine is an index of drug resistance.

Notes: RESUME. L' étude ultrastructurale de la grégarine Gregarina blaberae a été entreprise après mise au point d'un protocole d'infestation expérimentale de larves vierges de Blattes, Blaberus craniifer. Les observations en microscopie électronique, effectuées en fonction du temps, ont permis de suivre le développement du sporozoïte, ainsi que la croissance et l' évolution du trophozoïte.La croissance est spectaculaire puisqu'en 18 jours elle entraîne la transformation du germe infectieux ou sporozoïte (L = 15 μm, φ=1 μm) en un céphalin—trophozoïte fixéà l' épithélium intestinal—de 250 μm de longueur pour un diamètre de 65 μm. L'organisation ultrastructurale du sporozoïte ne diffère pas de celle de la plupart des sporozoïtes des autres sporozoaires étudiés jusqu' à présent, le conoïde et les corps denses sont présents. La paroi est trimembranaire. toutefois le complexe membranaire interne présente quelques interruptions. Les premiers dictyosomes s' élaborent à partir de l'enveloppe nucléaire. La migration du noyau et des corps denses, puis la régression de toutes les formations caractéristiques du sporozoïte et l' établissement d'une zone corticale venant coiffer l' épimérite, ont lieu dans les 48 heures suivant l'infestation et marquent la transformation sporozoïte-céphalin.L' évolution du céphalin, se poursuit par la formation des ébauches des plis épicytaires qui s'effectue à partir du 3éme jour du développement à la base du deutomérite. Un système régulier de plis longitudinaux ou épicyte s' établit ainsi sur toute la partie extra-cellulaire de la grégarine. Dès les 4éme-5éme jours de développement, un réseau vacuolaire et un chondriome important se différencient dans l' épimérite, tandis qu'un septum de nature fibrillaire isole le protomérite du deutomérite. Le stade suivant—à partir du 6éme jour—est marqué par une ségrégation du matériel glucidique au niveau de ces 2 segments.Le modèle étudié permet de préciser le rôle de l' épimérite dansla nutrition du parasite, l' évolution du chondriome et des membranes corticales au cours de la phase de croissance végétative des grégarines polycystidées.

Notes: SYNOPSIS. Stomatogenesis was studied in the heterotrich ciliate Blepharisma japonicum stained with protargol. During binary fission not only is a new oral apparatus made for the posterior daughter, but the already existing oral apparatus of the parent cell is reorganized, i.e. partially disassembled and then subsequently reassembled to provide a functional feeding apparatus for the anterior daughter cell. These morphogenetic events, requiring 21/2 to 3 hr, are complete by the time the anterior and posterior daughters separate.In preparation for division, an oral anlage is formed by the rapid proliferation of kinetosomes along 4–5 stomatogenic kinetics directly subtending the cytostome. This field of randomly oriented kinetosomes ultimately gives rise to the feeding apparatus of the posterior daughter cell. Early in division, the oral anlage separates into 2 longitudinal fields of kinetosomes: one is destined to give rise to the undulating membrane and the other forms the adoral zone of membranelles. Shortly after the anlage is established posterior to the cytostome, reorganization of the existing functional mouth is initiated. The morphologic changes associated with this dedifferentiation-redifferentiation sequence lead to the formation of an oral apparatus for the anterior daughter and cannot be distinguished from those characteristically seen during physiologic reorganization.

Notes: SYNOPSIS. The effects of age and sex of the cat on oocyst shedding, multiplication of Toxoplasma gondii in tissues of cats, and acquisition of immunity were investigated after oral inoculation of cats with Toxoplasma cysts. Twenty-five cats varying in age from 1 week to 39 months were killed 7-97 days after inoculation with T. gondii. Homogenates of brain, heart, mesenteric lymph nodes, retina, and blood from these cats were inoculated into mice to test for Toxoplasma infectivity. Toxoplasma was isolated more frequently and in higher titers in mice receiving inocula from cats of the youngest age group (1 week old). Toxoplasma gondii was isolated from tissues of only 2 of 21 cats older than 2 months (at the time of inoculation), although all of the animals shed oocysts within 1 week after ingesting the parasites.The number of oocysts shed varied among littermates of the same sex and between sexes. Generally, cats younger than 12 months shed more oocysts than older cats. The number of oocysts shed by older cats varied considerably; males generally shed more oocysts than the females. However, the numbers of cats examined were too small for statistical comparison. Nevertheless, the observations suggest that cats older than 12 months should not be used in experiments where numbers of oocysts shed is critical.

Notes: SYNOPSIS. Changes in the metabolism of Crithidia fasciculata ATCC 11745 when grown in the presence of ethidium bromide were studied. Ethidium bromide-grown cells had decreased respiratory activity as measured by oxygen consumption. More than 50% of the organisms cultivated in a defined medium containing 1.0 mg/liter of ethidium bromide became dyskine-toplastic and had decreased activities of particulate succinate and NADH-linked dehydrogenases as well as of soluble isocitrate dehydrogenase. These cells also had increased activities of particulate α-glycerophosphate dehydrogenase, soluble α-glycerophosphate dehydrogenase, malic enzyme, hexokinase, and malate dehydrogenase. Ethidium bromide-grown cells had a lower level of ATP and contained less DNA than cells grown in its absence.

Notes: SYNOPSIS. A video technic is described that permits a quantification of the degree of attraction of Trypanosoma cruzi trypomastigotes to vertebrate cells in vitro. Bovine embryo skeletal muscle cells (BESM), HeLa cells and Vero cells all attract a myotropic strain of T. cruzi trypomastigotes. BESM cells, however, are 2-fold more attractive to trypomastigotes than HeLa cells and 10-fold more attractive than Vero cells. Heat-inactivation of BESM cells abolishes their ability to respire and also to attract T. cruzi trypomastigotes. As there is no difference in the endogenous oxygen consumption between BESM, HeLa, and Vero cells, it is unlikely that differences in the attraction of trypomastigotes to the 3 cell types are due to variations in the magnitude of pO2 or pCO2 gradients in the milieu around the cells.

Notes: SYNOPSIS. Euglena gracilis strain Z has a motor response which results in orientation with respect to the polarization of a light stimulus. Cells swim preferentially in a direction perpendicular to the plane of polarization of the stimulus. If 2 polarized stimuli are given from opposite directions, the preferred direction is, under certain circumstances, at right angles to the directions of both stimuli. Euglena also preferentially assumes an orientation that is at right angles to the force of gravity. The relationships between these responses and phototactic movements oriented with respect to the direction of the stimulus are discussed.

Notes: SYNOPSIS. The behavior of Paramecium caudatum in small capillary glass tubes was investigated under various ionic conditions and at the various tube diameters. Along the inner walls of the tubes ciliates undergo regular spiral motion, which is completely different from natural spirallings or random walk-like movements observed usually in large vessels. The curvature calculated from the tracks of spiral motions was independent of the inner diameters of capillary tubes, but depend specifically on ionic conditions.A plausible law governing such regular spiral motions of Paramecium caudatum is proposed. A definite part of the anterior end of a ciliate seems to contact the curved surface of the inner wall of a capillary tube during the motion so that the organism receives a constant tactile stimulus, and the direction of motive force keeps a certain angle against the surface.

Notes: SYNOPSIS. Observations of the ultrastructure of marine scuticociliatids, tentatively assigned to the genus Uronema, were made by light, transmission electron, and scanning electron microscopy. Giant, cortically oriented mitochondria filled the subpellicular, intermeridional areas, and were in close association with the epiplasm immediately under the inner alveolar sac membranes. Reconstructions of serial sections of the posterior poles of ciliates indicated that the intermeridional mitochondria could fuse at that point and the entire chondriome might at times be a single organelle. A system of tubules was observed to be intimately associated with the mitochondria in the posterior region. The tubules anastomosed and were directed posteriorly into the region of the nephridial-contractile vacuole system. The outer surfaces were coated with projections arranged in helical patterns. The system may be regarded as a fluid segregation organelle. The tripartite nature of the polar basal body complex observed by silver impregnation was confirmed by transmission electron microscopy. The 3 structures were the basal body of the caudal cilium and 2 parasomal sacs. A prominent ring around the caudal cilium was observed by scanning electron micrcscopy; it is probably responsible for the silver deposition surrounding the polar basal body complex that can be seen by light microscopy of silver-impregnated specimens. The ultrastructure of the nonmotile caudal cilium and its kinetosome was unremarkable, being like that of the motile, somatic cilia. The micronuclear and macronuclear outer membranes were continuous at several sites. Such interconnections explain the intimate physical relationship between the nuclei during interphase in many ciliates, and could be a structural basis for chemical communication between the 2 nuclear types. Within the cytoplasm surrounding the opening of the cytoproct, numerous clear vesicles were observed. Their position and appearance suggested that the cytoproct may be involved in the elimination of solutions as well as solids. Food vacuoles, cortical microtubules, lamellar vesicles, disc-shaped vesicles, mucocysts, and a contractile vacuole and its pore were also observed.

Notes: SYNOPSIS. The occurrence and levels of activity of various enzymes of carbohydrate catabolism in culture forms (promastigotes) of 4 human species of Leishmania (L. brasiliensis, L. donovani, L. mexicana, and L. tropica) were compared. These organisms possess enzymes of the Embden-Meyerhof pathway but lack lactate dehydrogenase. No evidence could be found for the production of lactic acid by growing cultures and lactic acid could not be detected either in cell-free preparations or after incubation of cell-free extracts with pyruvate and NADH under appropriate conditions. All 4 species possess α-glycerophosphate dehydrogenase and α-glycerophosphate phosphatase which together could regenerate NAD, thus compensating for the absence of lactate dehydrogenase. The oxidative and nonoxidative reactions of the hexose monophosphate pathway are present in all 4 species. Cell-free extracts have pyruvate dehydrogenase activity which allows the entry of pyruvate into and its subsequent oxidation through the tricarboxylic acid cycle. All enzymes of this cycle, including a thiamine pyrophosphate dependent α-ketoglutarate dehydrogenase are present. Both NAD and NADP-linked malate dehydrogenase activities are present. The isocitrate dehydrogenase is NADP specific. There is an active glutamate dehydrogenase which could compete with α-ketoglutarate dehydrogenase for the common substrate (α-ketoglutarate). Replenishment of C4 acids is accomplished by heterotrophic CO2 fixation catalyzed by pyruvate carboxylase. All 4 species have high levels of NADH oxidase activity. Several enzymes thus far not found in any species of Leishmania have been demonstrated. These are: phosphoglucose isomerase, triose phosphate isomerase, fructose-1, 6-diphosphatase, 3-phosphoglycerate kinase, enolase, α-glycerophosphate dehydrogenase, α-glycerophosphate phosphatase, pyruvate dehydrogenase complex, citrate synthase, aconitase, α-ketoglutarate dehydrogenase, glutamate dehydrogenase, and NADH oxidase.

Notes: SYNOPSIS. Oocysts of Isospora vulpina were found in silver foxes (Vulpes vulpes) on a fox farm in Wisconsin. They were 29.7 (25-38) × 24.3 (21-32) μm. The sporocysts were 17.7 (15–23) × 13 (11–16) μm. Five coccidia-free puppies were inoculated with 22,000–42,000 oocysts each of I. vulpina from the fox: a patent infection resulted after 6-7 days. The infection was then transferred from 1 of these dogs to another coccidia-free puppy. After a 7-day prepatent period the puppy passed oocysts for 7 days.

Notes: 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.

Notes: 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.

Notes: 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.

Notes: 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.

Notes: 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.