ALBERT

Notes: . Dehydrogenase activity with hydroxysteroids has been observed in Tetrahymena furgasoni (formerly T. pyriformis strain W), and the enzyme responsible has been isolated from this organism. The purified dehydrogenase is active with a variety of steroid alcohols at apparent Km values ranging from 0.2 to 4.0 mM. The C-3 hydroxyl of ring A of the steroid nucleus is the preferred position of oxidation. However, a variety of other secondary alcohols are also substrates, with apparent Km values for 2-butanol, 2-pentanol, and cyclohexanol of 880, 1000. and 150 mM, respectively. With both steroidal and nonsteroidal alcohols. NAD is the preferred co-substrate, although low activity with NADP is observed. Evidence is presented that the activity with secondary alcohols, whether steroidal or not, is the property of a single protein species.

Notes: . Domestic turkeys naturally infected with Leucocytozoon smithi were blinded by bilateral ocular enucleation, pinealectomized, sham-pinealectomized, pinealectomized plus enucleated, or maintained as controls. Groups of turkeys were acclimated to either light-dark periods of 14L:10D or “darkness” with intermittent periods (10–20 min) of red light at irregular hours approximately every three days as required for maintenance of turkeys. Peripheral gametocyte numbers of L. smithi in all groups were determined every 2 h over a 36 h period. Under 14L: 10D photoperiod, no observable difference in the pattern of gametocyte circadian rhythmicity between pinealectomized, enucleated, pinealectomized plus enucleated, and control turkeys was noted. Although mean parasitemias differed among groups, peak gametocyte numbers occurred between 1000 and 1800 h; how parasitemias occurred between 2000 and 0400 h. However, the phase of gametocyte rhythmicity in pinealectomized plus enucleated turkey hosts did exhibit a lag with reference to other hosts when examined by least squares fits of simple harmonics. Under conditions of “darkness” with intermittent, irregular periods of red light, L. smithi gametocyte numbers of individual turkeys, pinealectomized, sham-pinealectomized, or maintained as controls, exhibited a circadian periodicity though parasite cycles were out of phase with the natural photoperiod to which the turkeys previously had been exposed. A slight drift out of phase of L. smithi gametocyte periodicity occurred among turkeys in the sham-pinealectomized and the control groups while a considerably more prominent drift out of phase was seen among the parasite rhythmicity patterns of the pinealectomized birds. Data indicate that the pineal gland of the turkey did not directly mediate L. smithi gametocyte circadian periodicity, although an indirect involvement in regulating the timing of parasite rhythmicity is suggested.

Notes: . The surface of merozoites and sporozoites of Eimeria tenella was affected by incubation with E. tenella-immune chicken serum (ICS). Normal chicken serum (NCS) and heat-inactivated ICS had no effect on the pellicular surface of either developmental stage. Sporozoites formed surface bulges or swellings after 10 min of incubation with ICS, and by 15 min postincubation, the morphology of the sporozoites was distorted by a surface coating of fibrinous material. Merozoites exposed to ICS were similarly coated, but surface swelling was not as severe. The coating formed rapidly and was seen as early as 5 min postincubation. Sporozoites incubated with heat-inactivated ICS supplemented with normal chicken serum were coated with a fibrinous material and in some cases lysed. These data indicated that complement must be present for the surface interaction to occur.

Notes: . The fine structure of the trophozoite, encysting cells, and the cyst of Acanthamoeba astronyxis has been examined. In the trophic form a microtubule organizing center was associated with a well developed Golgi complex. During encystment the organelles of the amoeba changed considerably. The profiles of rough endoplasmic reticulum elongated and were often arranged in circles of multilayered concentric systems, enclosing mitochondria, the nucleus, or other inclusions. The mitochondria showed a tendency toward elongation and constriction. One or two nucleolus-like bodies appeared in the nucleus. Lipid droplets increased considerably in amount and were distributed individually or as aggregates. The mature cyst was star-shaped and surrounded by an almost circular exocyst and an endocyst that was closely apposed to the cell membrane. Both walls differed in their thickness and granulation. The exocyst was continuous over the entire cyst, while the endocyst was interrupted by gaps, ostioles. in the region of the rays. Within the ostioles was a bell-shaped structure, the operculum. The latter was composed of a granular material comparable in electron density to that of the endocyst.

Notes: Ambiphrya ameiuri is an ectocommensal peritrich that attaches to the gills of warm-water fishes and filters bacteria from the water. The ultrastructure of this protozoon, its attachment to the fish gills, and its effect on the gill tissue were investigated by scanning and transmission electron microscopy. The peritrich attached to the gills by fibers extending from the scopula. A microtubular array, apparently a barren kinetosome, was present in each lobular projection, but no scopular cilia were observed. At low densities Ambiphrya had no apparent harmful effects on the fish; however, at high densities respiration may be impeded. Ultrastructural studies indicate that this organism receives no nourishment from the host tissue.

Notes: Une méthode d'obtention et de maintien des Ciliés du rumen (genre Polyplastron) en culture axénique, est décrite. Elle comporte deux périodes d'incubation en anaérobiose avec différentes associations d'antibiotiques, séparées par un lavage et un renouvellement du milieu nutritif. Dans ces conditions. Polyplastron multivesiculatum est obtenu à l'état axénique et maintenu en survie pendant cinq jours.〈section xml:id="abs1-2"〉〈title type="main"〉ABSTRACTA method to obtain and keep rumen ciliates (genus Polyplastron) in an axenic condition is described. It consists of two incubating periods with different antibiotic mixtures, separated by washing and renewing of nutrient medium. Under these conditions, Polyplastron multivesiculatum is established in an axenic state and can survive thusly for five days.

Notes: Proteins of whole cell extracts of Naegleria fowleri, precipitated with acetone, have been resolved by two-dimensional polyacrylamide gel electrophoresis. Autoradiograms of the [35S]-methionine-labeled polypeptides were scanned and analyzed by a computer-assisted program in order to determine whether there were correlations between selected attributes of proteins (e.g., subunit size and charge). The majority of the polypeptides had molecular sizes within the range of 20–60 kilodaltons. The mean amount of polypeptide was less for those with molecular sizes between 20 and 45 kilodaltons than for those larger than 45 kilodaltons. The mean amount of polypeptide was greater in the isoelectric focusing range of pH 5–6 than in the range of pH 6–7. Polypeptides in the size range of 20–40 kilodaltons had a median isoelectric point of 6.1, whereas polypeptides in the size range of 40–80 kilodaltons had a median pI of 5.6. Our data indicated that molecular size and charge were not entirely independent variables, and that the composition of a polypeptide might have an important influence on its steady state level in N. fowleri.

Notes: During a survey of the coccidian parasites of reptiles from Iowa, three specimens of Crotalus horridus L., the Timber Rattlesnake, and one of Sistrurus catenatus (Rafinesque), the Massasauga Rattlesnake, were found to be passing oocysts of a Caryospora, here described as C. bigenetica n. sp. Since these snakes (family Crotalidae) are known to subsist mainly on small mammals, oocysts from one of the Timber Rattlesnakes were fed to laboratory white mice (Mus musculus L.) to determine if mammals might be involved as alternate hosts in the life cycle. At necropsy, tissues of the tongue and dermis of the mice revealed a sequence of stages which included mature male and female gamonts, fully sporulated sporocysts, “excysted” sporozoites, and “resting” sporozoites that lay individually in solitary, cyst-like host cells termed “caryocysts.” A coccidia-free Massasauga that was fed an infected mouse, at a time when caryocysts in the mouse would have been present, later passed oocysts similar to those of the original inoculum. These results, along with the discovery of endogenous stages (asexual and sexual) in the intestine of the Timber Rattlesnake and the experimentally infected Massasauga, suggest that this parasite has a heteroxenous life cycle pattern, with sexual stages occurring both in the ophidian and the mammalian hosts.

Notes: Ciliophrys marina is a small marine helioflagellate, with a central nucleus, which is capable of reversibly transforming from a rapidly swimming flagellate cell with no axopodia to the structure of a heliozoan with a flagellum that beats only a few times a minute. When in the flagellate form, the flagellum acts as a tractellum due to the tubular mastigonemes found along its length. When the rapidly swimming flagellate strikes a piece of debris, the flagellum goes through a very characteristic shock-induced avoidance reaction. Similarly, when a mechanical shock is delivered to the cell in its heliozoan form, the axopodia are contracted in less than 20 msec. Both reactions are inhibited in low calcium seawater. Transformation from the heliozoan to the flagellate form is accomplished by slow retraction and absorbance of the axopodia and activation of the flagellum. Ultrastructurally, each axopodium is found to contain three microtubules which attach to the outer nuclear membrane of the central nucleus at sites that this study characterizes by electron microscopy of thin sections and freeze fracture preparations. The mitochondria have tubular cristae, each containing an intracristal filament. Finally, a taxonomic review of the helioflagellates is presented, and it is suggested that C. marina is derived from the chrysomonads. An argument is also made for classifying C. marina with the heliozoan order Actinophryida, as a recently published classification of the protozoa does.

Notes: Laboratory-reared Fundulus grandis and F. heteroclitus were experimentally infected with Eimeria funduli by being fed Palaemonetes pugio (grass shrimp) collected from endemic areas. Histological sections were made of heart, liver, hepatopancreas, spleen, gall bladder, kidney, intestine, peri-intestinal fat, reproductive organs, and brain from F. grandis sacrificed at 1, 2, 6, 12, 18, and 24 h and from F. heteroclitus at 5, 6, 9, 14, 19, 24, 29, 34, 39, and 44 days after consuming naturally infected shrimp. We first found merogonous stages at day 9 postinfection (p.i.). No developmental stages of the parasite could be positively identified in the tissues of experimentally infected fish prior to day 9 p.i. Mature meronts were found 14 days p.i. The majority contained 8–16 (mean, 13) merozoites, but a few meronts had 18–26 (22) merozoites. Gamonts first appeared on day 14, were mature by day 19, and fertilization was completed by day 24 p.i. After sporoblast formation, sporopodia appeared during sporocyst wall formation, between days 24 and 29 p.i. Sporozoite formation was completed by day 44 p.i. in most sporocysts. Most endogenous stages occurred in hepatocytes; however, pancreatic and spleen cells were sometimes infected with gamonts.

Notes: A recent isolate of Eimeria praecox, strain G, was obtained from Georgia and purified. Studies of the life history, pathogenicity, and cross-immunity of the isolate were conducted to verify its identity. In inoculated three-week-old chickens, the occurrence of merogony and gametogony was limited to the superficial epithelium of the upper intestine. Oocysts, 23 × 19.5 äm, with a shape index of 1.17 were first observed 83 h after inoculation. Mortality and morbidity were not observed in any of the experimental birds. However, there was a positive correlation between dose of oocysts, reduced weight gain, and the incidence of exudative diathesis. These studies showed that E. praecox depresses weight gains in chickens and may be of economic importance. Although complete immunity to avian coccidiosis is believed to be species specific, chickens immune to E. praecox (G) or E. acervulina had a degree of cross-immunity to a heterologous challenge. Electrophoretic analysis of glucose phosphate isomerase and lactate dehydrogenase prepared from the European strain of E. praecox and E. praecox (G) showed no differences, confirming the identity of the isolate as E. praecox.

Notes: The antiviral agent phosphonoacetic acid inhibits growth of Tetrahymena thermophila at concentrations comparable to those inhibiting growth of other eukaryotic cells, with 50% inhibition at 0.5 mM phosphonoacetic acid. The compound is cytotoxk to Tetrahymena at concentrations greater than 2.0 mM. When a culture of Tetrahymena the growth of which was totally inhibited by 2.0 mM phosphonoacetic acid was diluted with fresh medium, growth resumed in an exponential, rather than synchronous, fashion. [2–14C]phosphonoacetic acid is not metabolized by Tetrahymena.

Notes: Agglutinins were not detected in sera from mice given one, two, or three intranasal (i.n.) inoculations or a single intravenous (i.v.) inoculation of trophozoites of Naegleria fowleri. However, agglutinins were produced following second and third i.v. inoculations. Serum immunoglobulin levels increased in both i.n.- and i.v.-inoculated mice. IgG and IgM increased substantially more for i.v.-inoculated mice. IgA levels increased more consistently for i.n.-inoculated mice.

Notes: The sensitivity and specificity of the indirect fluorescent antibody (IFA) test for the detection of serum antibodies were examined in mice that were infected with Eimeria falciformis, E. ferrisi, E. papillata, or E. vermiformis. For the study of each species, five groups of mice were given graded inoculation doses of 10, 102, 103, 104, or 105 sporulated oocysts in a primary infection. The sixth group was infected with three sequential doses of 1.5 times 103, 1.5 times 104, and 1.5 times 105 sporulated oocysts per mouse at two- to three-week intervals. All groups of infected mice developed serum antibodies. Sera were titrated by the IFA test with purified sporozoites. Strong fluorescence and high IFA titers were observed with homologous reactions mainly with the sera from mice infected with the higher inoculation dose levels in primary infections and from those given three sequential inoculation doses. Immunological cross reaction among the four species of Eimeria occurred at dilutions of 1:10 to 1:160. Very weak or no fluorescence of free sporozoites was observed with sera from noninfected mice, and there was no fluorescence of sporozoites contained in intact sporocysts.

Notes: The structure of both the host and parasite membranes during stages in the asexual development of Plasmodium chabaudi in mouse red blood cells is examined by transmission electron microscopy of thin sections and freeze-fracture preparations. The erythrocyte's plasma membrane, the membrane of the parasitophorous vacuole, and the plasma membrane of the parasite exhibit different structural properties in terms of membrane width and the frequency and diameter of the typical intramembrane-particles (IMP) populating the membrane's fracture faces. The difference between the parasitophorous vacuolar membrane and host cell's plasma membrane is remarkable because the vacuolar membrane is formed from an invagination of the erythrocyte's plasma membrane. The vacuolar membrane has significantly reduced frequencies and diameters of IMP's on both faces and reveals a marked temperature response manifesting itself as large IMP-devoid domains emerging on both faces on chilling to 4°C. In contrast, cooling induces only some very small IMP-devoid patches on both faces of the host plasma membrane. Neither of these membranes changes significantly as parasite development progresses. In contrast, the parasite's plasma membrane shows local alterations during its development, forming compaction domains with the nuclear envelope in ca. 30% of the ring-stages and trophozoites. These compaction domains disappear in late uninuclear trophozoites and schizonts. Furthermore, the plasma membrane of the host cell, the vacuolar membrane, and the parasite's plasma membrane do not respond to externally applied Ca2+, and their temperature-response remains unaltered during the infection cycle. Thus, modification of these three membranes as a consequence of invasion and development of the parasites, as recently found in the primate malaria caused by P. knowlesi, can be detected neither directly nor indirectly via temperature- and/or Ca2+-response in the rodent malaria caused by P. chabaudi.

Notes: A system of prescreens and screen has been developed to select chelators as potential drugs against Trypanosoma brucei brucei EATRO 110. The chelators tested were nearly all commercially available, low molecular, and having moderate to high affinity for Fe(III). We prescreened 70 compounds showing heme-sparing or inhibitory activity in a Crithidia fasciculata growth system having excess Fe and minimal hemin. Of these, 45 were highly trypanocidal for suspensions of bloodstream T. b. brucei; criteria of activity here were immobilization, lysis, and loss of infectivity. Eighteen of the chelators highly active in the suspension prescreen were tried in T. b. brucei-infected mice. Thirteen of these chelators were curative in mice with 24-h infections, that is, they allowed survival 〉30 days beyond the untreated controls. 3,4-Dihydroxycinnamic acid (caffeic acid). 2,9-dimethyl-1, 10 phenanthroline (neocuproine), and 2-pyridinecarboxaldehyde-2-pyridyl-hydrazone cured five out of five mice after an i.v. dose of 100 mg/kg. Salicylaldehyde thiosemicarbazone cured five out of five mice at an i.p. dose of 500 mg/kg. Lesser activity was shown by several other chelators.

Notes: The size and fatty acid composition of Tetrahymena pyriformis W cells were influenced by the provision of a nutritional supplement of ergosterol, cholesterol, or tetrahymanol, but not of 20-isocholesterol. Ergosterol and cholesterol addition led to a reduction in cellular volume, an increase in glycerophospholipid saturated fatty acid content, and an increase in palmitoleic acid and its metabolic products when compared to unsupplemented controls. Tetrahymanol supplementation resulted in an increase in cellular volume, a decrease in saturated fatty acid content, and a reduction in palmitoleic acid and derivatives. 20-Isocholesterol was accumulated by the cells; however, this compound had no effect on any of the parameters followed in this investigation and had only a small depressant effect on tetrahymanol biosynthesis. Ergosterol and cholesterol had the same impact on the ciliates, even though the ergosterol-supplemented cells contained approximately three times as much free sterol as did cholesterol-grown cells. The amount of the free cholesterol and metabolic products in supplemented cultures was similar to the amount of tetrahymanol present in control cultures. This observation suggests that the cells recognize qualitative differences among the various polycyclic alcohols rather than responding to the amount of sterol present. Increased cellular levels of tetrahymanol led to a response unlike that of the true sterols, which again suggests that the high degree of specificity depends on the structure of the added polycyclic alcohol. The changes in fatty acid composition may be required to maintain proper interaction of the polar lipids and the polycyclic alcohols to give an appropriate degree of membrane fluidity.

Notes: An unknown, ninhydrin-positive substance detected on paper chromatograms of the endogenous metabolites of mixed rumen ciliate protozoa was isolated and purified by column chromatography with ‘Dowex’ 50-X8 resin and identified as 2-aminobutanoic acid (α-amino-n-butyric acid) on the basis of elementary analysis, mass spectrometry, paper chromatography, infrared spectrometry, and melting point.

Notes: Book reviews in this article: Dillon, Lawrence S. 1981. Ultrastructure, Macromolecules and Evolution Schwemmler, Werner & Schenk, Hainfried E. A., eds. 1980. Endocytobiology: Endosymbiosis and Cell Biology, a Synthesis of Recent Research Krylov, M. V. & Starobogatov, Y. I., eds. 1980. Principles of the Construction of the Macrosystem of the Unicellular Animals Palmer, W. J., ed. 1981. Rapid & Automated Methods in Microbiology & Immunology: A Bibliography, 1976–1980 de Harven, E. & Nemanic, M. K., organizers. 1981. Cell Surface Labeling Inoki, Shozo, ed. 1981. Atlas of Protozoa Czapik, Anna 1980. Pdostawy Protozoologii. [Introduction to Protozoology.] Levine, Norman D. & Ivens, Virginia 1981. The Coccidian Parasites (Protozoa, Apicomplexa) of Carnivores Page, Frederick C. 1981. The Culture and Use of Free-Living Protozoa in Teaching

Notes: SYNOPSIS. Traditionally, observations on the nature of protozoa have been published in periodicals or books, or remain buried in research notebooks. The retrieval and processing of information on a particular species or strain are dependent solely upon individual investigators. Although various modern methods have been applied to the study of protozoa, no attempt has been made to develop a system with which information on protozoan strains can be stored, retrieved easily, and processed for various analyses by computer technology. Based upon an existing system for encoding data on bacterial strains, a complementary system applicable to protozoan strains was developed and is described herein.

Notes: SYNOPSIS. The development of Sarcocystis cruzi Hasselmann (syn. S. fusiformis Railliet) meronts was studied in seven 7- to 10-day-old calves killed 4, 7, 11, 15, 22, 25 and 28 days postinoculation (DPI) with 5 × 107 sporocysts from feces of coyotes. No meronts were found 4 and 7 DPI. Young and intermediate meronts with 1–16 nuclei were found in endothelial cells of arteries in mesenteric lymph nodes, but not in kidneys 11 DPI.Mature meronts were noted in endothelial cells of arteries, arterioles, or capillaries of many organs of calves killed 15 to 25 DPI. No first-generation meronts were found 28 DPI. By electron microscopy, all stages of the first-generation merogony were found free within the host cell cytoplasm and not within a parasitophorous vacuole. The appearance of intranuclear spindles preceded the formation of merozoites by endopolygeny. Mature meronts measured 41.0 × 17.5 (34–50 × 15–24) μm, contained ∼ 100–350 merozoites, and had 2 to 4 relatively small residual bodies, 2.8 μm in diameter. Merozoites measured 6.3 × 1.5 (5.5–7 × 1 μm) and contained most of the organelles characteristically found in coccidian merozoites. Micropores were observed in merozoites, but not in young and intermediate meronts. Merozoites were seen free in the lumen of blood vessels, in intracellular areas, and free within the host cell cytoplasm.

Notes: SYNOPSIS. Myxidium spores from various eel hosts (Anguilla spp.) are compared. Myxidium anguillae, M. enchelypterygii, M. illinoisense, M. serum, and M. zealandicum are synonymized with M. giardi, a ubiquitous species reported from A. anguilla, A. rostrata, A. mossambica, A. japonica, A. reinhardtii, A. bicolor pacifica, A. australis, and A. dieffenbachii. Myxidium uchiyamae, M. lentiforme, M. matsuii and M. acinum are retained, and 2 new species described. Species other than M. giardi appear to be restricted to the Indo-Pacific region.

Notes: SYNOPSIS. A new species of kinetophragminophoran ciliate, collected from dried vegetation and capable of forming an aerial sorocarp, is described and named Sorogena stoianovitchae gen. n., sp. n. This ciliate is a voracious predator that feeds on species of Colpoda, and, when the latter is depleted in numbers, aggregates to forms sorogens. Each sorogen rises into the air from the surface of the water, forming a secreted stalk with a sorus of cysts at its apex. the feeding stage of the ciliate resembles an Enchelys in that it has an apical, slit-like mouth surrounded by a lip, a somewhat dorso-ventrally flattened body, and meridional kineties. Its length ranges from 40–75 μm and width from 23–55 μm. It has a typical rhabdos type of cytopharynx, but no specialized oral ciliature. the somatic kineties are formed of rows of paired kinetosomes with associated microfibrils, the arrangement of which differs a little from that of other ciliates of this subclass. Sorogena has tentatively been placed in the order Haptorida although it lacks toxicysts, recognizable mucocysts, and clavate cilia. Its unique life cycle and some of the details of its fine structure indicate differences between Sorogena and other haptorids so profound that a new family, SOROGENIDAE, is created for it. the type species (PNG76-73) was collected on dry figs at the Wau Ecology Institute, Papua New Guinea.

Notes: Cell surface pellicular membranes (PM) were isolated from promastigote forms of Leishmania donovani by differential and discontinuous sucrose gradient centrifugation procedures. the PM had a density equivalent of ∼ 1.19 g/cm3. As ascertained by electron microscopy, longitudinal parallel arrays of subpellicular microtubules (MT) remained attached to the isolated PM inner lamina, and this feature was used to assess membrane fraction purity. Gradient fractions having ∼ 95% of all membranes combined with MT were obtained routinely. the attached MT imparted a structural asymmetry to the PM permitting uniequivocal identification of the membrane external and cytoplasmic surfaces. the supramolecular structure of attached MT was evident in negatively stained PM. In ultrathin sections, PM had a mean width of ∼ 7.2 nm and attached MT a diameter of ∼ 29 nm. the MT were apparently cross-bridged both to each other and to the PM via a flocculent filamentoid nexus. As determined by sodium dodecylsulfate-polyacrylamide gel electrophoresis, isolated PM contained ∼ 40 peptide bands ranging in apparent molecular weight from ≤ 1.2 × 104 to ≥ 2.2 × 105daltons. of these, 19 were stained with periodic acid-Schiffs’ reagent suggesting that most PM carbohydrate constituents were present as glycopeptides. A presumpative glycolipid/polysaccharide PM constituent was also identified in such gels.

Notes: Organic requirements for attachment to glass, elongation, and motility of Entamoeba histolytica, have been determined. the trophozoite, which has been grown axenically only in highly complex media with reduced oxygen tensions, remains rounded and detached when placed in a Tris-HCl buffered solution containing NaCI, KCI, MgCI2, and CaCI2. A maintenance medium in which the amebae could attach to glass, elongate, and remain motile and viable for 12 to 24 h was devised with the addition of cysteine, ascorbic acid, bovine serum albumin, and the vitamin solution of medium NCTC #107. Tris-HCI was the most effective buffer tested and the optimal pH was 6.9 to 7.0. Survival, but not attachment, of the amebae was decreased at osmolalities ranging between 110 and 180 milliosmoles/kg, whereas both functions were decreased above ∼260 milliosmoles/kg. Bovine serum albumin, the most effective of the proteins tested, and the vitamin solution helped maintain attachment of some ameba strains, but were not required by other strains. the requirements for cysteine and ascorbic acid were absolute and highly specific. During incubation in the maintenance medium, cell volumes decreased. Sensitivity of the organisms to agglutination by concanavalin A, wheat germ agglutinin, soybean agglutinin and fucose binding protein remained unchanged.

Notes: SYNOPSIS. Differences in the composition and distribution of cell membrane carbohydrates were demonstrated in the 3 life cycle forms of 3 Trypanosoma cruzi strains by using lectins with different specificities. The results suggest that lectin binding may be useful in characterization of the parasite strains.

Notes: SYNOPSIS. Promastigotes of Leptomonas sp., a flagellate parasite of the silkworm, Bombyx mori, multiplied by binary fission with the following sequence of events: duplication of the flagellum; division of the kinetoplast and the nucleus; spatial separation of the kinetoplast: and cytokinesis resulting in the formation of 2 daughter promastigotes. In the early stages of encystment, promastigotes aggregated in a rosette and assumed a stumpy form. The nucleus and kinetoplast of the stumpy promastigotes were double, suggesting a possibility of fusion of the organism in the rosette. When most of the promastigotes in the cluster became stumpy, each individual was isolated from the cluster and acquired a thick coat with an acidophilic substance, thus forming a cyst.

Notes: Book reviewed in this article:Gantt, Elisabeth, ed. 1980. Handbook of Phycological Methods. Developmental and Cytological Methods.

Notes: SYNOPSIS. Doublet Paramecium tetraurelia would be expected to contain 2 macronuclei if their nuclear complement were strictly analogous to that of singlets. However, most doublets are unimacronucleate. It is shown in this study that dimacronucleate cells are present only in young clones. Unimacronucleate cells arise either through abnormalities in the determination and distribution of macronuclear anlagen during the first cell cycle after conjugation, or from dimacronucleate cells through abnormal division and segregation of macronuclei during the fission process.When a change in the number of macronuclei occurs through abnormalities in the division and segregation of daughter macronuclei, the daughter cells produced typically have DNA contents more similar than those expected from either random segregation of daughter macronuclei, or from the normal segregation pattern in ciliates in which changes in the number of macronuclei in progeny cells do not occur. This suggests that part of the regulation process of macronuclear DNA content in Paramecium may occur through control of the segregation pattern of daughter macronuclei.

Notes: SYNOPSIS. The relation of humoral antibody response to protection was evaluated in mice immunized with whole homogenates of Trypanosoma cruzi or with its flagellar fraction by direct agglutination and indirect fluorescent antibody test as well as by lytic and neutralizing activity against blood trypomastigotes. The results indicated that lytic antibodies were not implicated directly in protection against these trypanosomes. It was evident from histopathologic examination that the higher the degree of protection achieved, the lower the tissue damage observed in the challenged mice. Serum-neutralizing activity was highest in the groups protected most effectively.

Notes: SYNOPSIS. We have examined various properties of DNAs from 7 dinoflagellate isolates of wide geographic distribution; all of the isolates are superficially indistinguishable from a laboratory strain of Crypthecodinium cohnii originally isolated at Woods Hole, Massachusetts (WHd strain). Two isolates, one from Puerto Rico and the other from Honduras, are clearly distinguishable from WHd and the other isolates by their DNA buoyant density values. WHd and the other 5 isolates we have examined are indistinguishable from one another in terms of DNA buoyant densities and melting temperatures. The relationship among the various isolates, including WHd, were evaluated at a finer level through restriction endonuclease cleavage and molecular hybridization to compare ribosomal RNA gene structure in the several DNAs. All the isolates could be further categorized by this method, the patterns of restriction endonuclease cleavage of ribosomal RNA genes in the isolates paralleling exactly their sexual compatibilities established from breeding experiments by Beam & Himes. The DNAs were also treated with a restriction endonuclease sensitive to the presence of the modified base 5-methylcytosine. In all isolates, cytosine residues in both total DNA and DNA specifically containing the ribosomal RNA genes were found to be extensively methylated, as was previously shown for the WHd strain.

Notes: SYNOPSIS. Chemical procedures remove some of the outer 3 limiting membranes of 2 ciliate protozoa, Euplotes eurystomus and Tetrahymena pyriformis, and reveal sheets of microtubules in their ectoplasm for SEM study. This greatly enhances the analysis of the 3-dimensional geometry of these sheets, as is shown especially for E. eurystomus. In this organism, sheets of microtubules can readily be observed and described as they course through or around parts of the oral apparatus and other 3-dimensionally complex regions.

Notes: SYNOPSIS. A large, external glycoprotein with antigenic properties isolated from the ciliate Pseudomicrothorax dubius was found to have a molecular weight of ∼ 250,000 daltons. Analysis of the extracts by isoelectric focusing in combination with immunodiffusion and gradient polyacrylamide gel electrophoresis revealed that the principal antigen was a large glycoprotein. the glycoprotein was purified partially by Sephadex ultrafiltration. and almost completely by affinity chromatography on a concanavalin A-Sepharose column.

Notes: SYNOPSIS Nosema algerae, a microsporidan parasite of anopheline mosquitoes, was successfully replicated in 3 insect cell culture lines: Trichoplusia ni (TN-368); Heliothis zea (IPLB-1075); and Mamestra brassicae (IZD-Mb-0503). Infectious spores were produced in vitro. Spores were observed at 48 h postinfection, and some cells were filled with sproes by 72 h.The number of parasites per cell increased with time. At 72 h postinfection, the infection rates for the 3 cell lines ranged from 23 to 32%. Infected cell lines were subcultured, and by the 6th passage spore production had ceased.

Notes: The somatic and oral ultrastructures of Woodruffia metabolica Johnson & Larson are described. the somatic kinetids are dikinetids, with the anterior kinetosome often not ciliated. A transverse ribbon of microtubules and a single postciliary microtubule are associated with the anterior kinetosome. A transverse ribbon, postciliary ribbon and kinetodesmal fibril are linked with the posterior kinetosome. the posterior transverse ribbons extend posteriad, to the left of the Kinety, joining more anteriorly originating transverse ribbons in a compound LKm fiber. These features together with interkinetosomal linkages relate Woodruffia to other members of the order Colpodida.

Notes: Developmental stages of Eimeria meleagrimitis Tyzzer were found throughout the intestine and ceca of turkeys given inocula ranging from 104 to 7.5 × 105 sporulated oocysts/bird. Infection initially occurred in the duodenum and upper jejunum but later moved down the intestine and into the ceca. the speed with which the infection moved into these areas was roughly proportional to the inoculum size. Heaviest infections were in the ileum, neck of the cecum, and large intestine. the life cycle consisted of 5 asexual generations before gametogony, a 6th asexual generation developing simultaneously with gametogony. First- and 2nd-generations were located along the sides of villi in the upper intestine rather than in the crypts of Lieberkühn, as previously described in England for this species. Transitory first-generation stages that were abnormally large and usually degenerate were found in the neck of the cecum.

Notes: Galactosephilic and mannosephilic lectins from Pseudomonas aeruginosa interact with Tetrahymena pyriformis GL. Specific adsorption of these lectins onto the Tetrahymena can be shown by inhibition of hemagglutination and by peroxidase binding to the cells mediated by the mannosephilic lectins. Interaction with the lectins does not agglutinate the protozoa even after immobilization by Na fluoride, formaldehyde, and glutaraldehyde or after papain treatment. However, inclusion of the lectins in the growth medium increases the growth rate of Tetrahymena and their presence in the medium supplied to starved ciliates increases phagocytosis of Chinese ink and vacuolization.

Notes: Nineteen clones of Trypanosoma cruzi were obtained as single-cell isolates from Triatoma infestans. Ten of the clones were isolates from a patient with chronic Chagas' disease; nine clones were isolates from a dog infected with T. cruzi strain CA-I isolated originally from a chronic chagasic patient. The growth kinetics and peak modal Coulter volume of these clones were characterized. Significant inter- and intra-group differences between growth rates and peak modal volumes were found. These data indicate that subpopulations and, consequently, genetic heterogeneity of T. cruzi exist in chronic chagasic patients. All of the clones infected vertebrate cells in vitro.

Notes: The ox-coyote cycle of Sarcocystis cruzi was studied by killing 38 calves between 4 and 153 days postinoculation (DPI) with 55 × 103-5 × 108 sporocysts from the intestines of coyotes. At 4 DPI, a zoite was found within the lumen of a mesenteric lymph node artery. At 7 DPI, zoites were found in mononuclear cells and in endothelial cells in mesenteric arteries. First generation meronts (41.0 × 17.5 μm in diameter) occurred 7–26 DPI in mesenteric lymph nodes. At 19–46 DPI, second generation meronts occurred in kidneys, muscles, and other tissues: renal meronts were 19.6 × 11.0 μm, and intramuscular meronts were 25.0 × 11.1 μm. Merozoites were found in the peripheral blood 17 DPI and later at 24–46 DPI. They divided by endodyogeny in mononuclear cells. Sarcocysts were seen first in the heart at 45 DPI and contained one or two metrocytes. At 55 DPI, sarcocysts containing only metrocytes were found in striated muscles, heart, and in smooth muscles of the urinary bladder, rumen, omasum, abomasum, and small intestine. At 67, 87, 112, and 153 DPI, sarcocysts were found only in striated muscles and in the heart. At 67 DPI, sarcocysts were up to 360 μm long. They contained only metrocytes and were not infective to the dog. At 86 DPI, sarcocysts contained mostly bradyzoites, a few metrocytes, and were infective to a coyote. The thin-walled sarcocysts grew to a maximum length of 800 μm and contained bradyzoites that were 10.9 × 3.0 μm. At 90 DPI, two mature sarcocysts were found in 2 of 73 sections of brain and spinal cord; hundreds of sarcocysts were present in sections of tongue and heart of this calf. Gametogony occurred in the small intestine of the coyote. Macro-and microgamonts were found in goblet cells of the small intestines of coyotes 6 h after the ingestion of infected meat. Microgamonts were few and contained 3–11 slender gametes. Oocysts were seen at 12 h and sporulation was completed 9 DPI. The prepatent period in the coyote was 8 days. The ox-coyote cycle is compared with ox-dog cycle.

Notes: The structure of the oral apparatus in the carnivorous macrostomal form of Tetrahymena vorax has been investigated using serial thin sections and preparations of isolated oral apparatuses. The cilia of the oral apparatus are organized into an undulating membrane that borders the right and part of the posterior margin of the buccal cavity and three membranelles that project from plateaus on the anterior surface. Each membranelle consists of one short row and two longer rows of hexagonally packed kinetosomes. The organization of the microtubules of the oral ribs is identical to that in the T. vorax microstomal cell type. However, the first oral rib originates near the first kinetosome at the anterior end of the undulating membrane. The fine filamentous reticulum that underlies part of the oral ribs in the macrostomal cell type is not striated, unlike the reticulum in the microstomal form. A band of filaments similar to the fine filamentous reticulum extends around the anterior margin of the large cytostomal opening that occupies most of the posterior part of the oral cavity. The single row of microtubules along the left side of the oral cavity and cytostome also has filaments associated with it. A major difference between the microstomal and macrostomal forms in the structure of the oral apparatus is in the oral connectives. The macrostomal cell type contains only a single cross-connective that joins the three membranelles and the anterior portion of the undulating membrane. The posterior or peripheral connective between the posterior ends of membranelles one and two and the posterior end of the undulating membrane is absent.

Notes: . Studies of the bristle (dorsal) cilia of Euplotes minuta. E. aediculatus, and Stylonychia mytilus by light and electron microscopy indicate that these cilia do not beat metachronously in any of the species. The bristle cilia in Stylonychia may beat actively, but those in Euplotes stand erect or are bent in different directions with the flow of water. The duration and degree of bending appear correlated with the duration and velocity of the water current. The fine structure of the bristle complex is similar in both Euplotes species and like other reports of Euplotes in the literature. The complex consists of paired kinetosomes, the anterior bearing a short cilium containing four to six rows of fibrous balls (lasiosomes) oriented along the anterior surface of the axoneme, the posterior lacking a cilium but with a small cap. Microtubular ribbons are associated with the paired kinetosomes, and a collar with a pronounced alveolar ring underneath the pellicular membrane tightly surrounds the cilium at the opening of the bristle pit. The bristle complex in S. mytilus differs from that of Euplotes and other hypotrichs in that it has a single kinetosome in interphase cells and, attached to the kinetosome, a prominent fibrous structure (parakinetosomal body). Microtubules are attached to the parakinetosomal body. As in Euplotes, the bristle unit is surrounded by mucocyst-like organelles (ampules). Observations of behavior and fine structure suggest that the dorsal bristles may be sensory, perhaps responding to stimuli from water currents, although other functions are possible, too.

Notes: . The sequence of formation and ciliation of basal bodies and the subsequent organization of compound ciliary structures of the oral apparatus of Tetrahymena thermophila was reanalyzed with the aid of scanning electron microscopy of cells in which the epiplasmic layer was exposed, as well as by light microscopy of protargol-impregnated specimens. This combination of methods allowed the delineation of numerous steps in the patterning of the oral ciliature, some of which have received little or no previous attention. Highlights include: the initial formation of “strings” of nonciliated new basal bodies in juxtaposition to relatively few basal bodies of the stomatogenic kinety; generation of basal body pairs, roughly oriented along the anteroposterior axis of the cell, that later align side-by-side to assemble promembranelles; condensation and reorientation of promembranelles simultaneous with addition of a third row of basal bodies anterior to the original two rows; production of a very short fourth row of basal bodies at the anterior right end of each developing membranelle; generation of the outer basal body row of the undulating membrane (UM) after alignment of the inner row, with transient ciliation of the inner row preceding permanent ciliation of the outer row; limited basal body resorption at the ends of membranelles; and sculpturing of the right ends of membranelles by a movement of basal bodies associated with formation of the ribbed wall adjacent to the UM. In the old anterior oral apparatus a repetition of the processes of generation of a new outer UM row and sculpturing of right ends of membranelles takes place in synchrony with the corresponding events in the oral primordium, following prior shedding of the old outer UM row and loss of the sculptured pattern in association with temporary regression of the ribbed wall micro-tubules. Oral development is complex, with different processes involved in the assembly of the membranelles and the UM, and with a sequence of distinct events involved in the generation of each of these structures. Speaking comparatively, membranelle development follows the same pathway in many, perhaps all, ciliates in which these structures or their homologues develop from a common stomatogenic field.

Notes: . The temporal changes in the size and pH of digestive vacuoles (DV) in Paramecium caudatum were reevaluated. Cells were pulsed briefly with polystyrene latex spheres or heat-killed yeast stained with three sulfonphthalein indicator dyes. Within 5 min of formation the intravacuolar pH declined from ∼7 to 3. With the exception of a transient and early increase in vacuolar size, vacuole condensation occurred rapidly and paralleled the acidification so that vacuoles reached their lowest pH and minimal size simultaneously. Neutralization and expansion of vacuole size began when vacuoles were GT8 min old. No labeled vacuoles were defecated prior to 21 min after formation but almost all DV were defecated within 1 h so that the digestive cycle of individual vacuoles ranged from 21 to 60 min. Based on these size and pH changes, the presence of acid phosphatase activity, and membrane morphology, digestive vacuoles can be grouped into four stages of digestion. The DV-I are GT6 min old and undergo rapid condensation and acidification. The DV-II are between 4 to 10 min old and are the most condensed and acidic vacuoles. The DV-III range in age from 8 to ∼20 min and include the expanding or expanded vacuoles that result from lysosomes fusing with DV-II. The DV-IV are GD21 min old, and since digestion is presumably completed, they can be defecated. The rise in intravacuolar pH that accompanies vacuole expansion suggests that lysosomes play a role in vacuole neutralization in addition to their degradative functions. The acidification and condensation processes in DV-I appear to be unrelated to lysosomal function, as no acid phosphaiase activity has been detected at this stage, but may be related to phagosomal functions important in killing food organisms, denaturing proteins prior to digestion, and preparing vacuole membrane for fusion with lysosomes.

Notes: . Total DNA/organism was determined by flow cytometry on stocks of 33 single-cell-isolate clones and one strain of mithramycin-stained Trypanosoma cruzi. Interstrain differences in mean total DNA/group of 34% and interclone differences in total DNA/organism of 41% were found. Microspectrofluorometric analyses of the trypomastigote stage of selected clones confirmed the flow cytometry data and indicated that the total DNA/organism differences were due to differences in DNA of both the nucleus and kinetoplast with the nucleus being the major contributing factor. These data imply that the potential for genetic diversity in T. cruzi may be very large.

Notes: . The cortical membrane proteins of three gregarine and one coccidian species were compared using sodium dodecyl sulfate/polyacrylamide-gel electrophoresis. About 30 proteins were identified in the ghosts of Gregarina blaberae and G. garnhami and 20 in G. rhyparobiae ghosts and Sarcocystis tenella pellicles. No protein with the electrophoretic mobility of muscular actin was present in the ghosts of the sporozoan species under study. Each species possessed a characteristic electrophoretic pattern; no protein was present simultaneously in the four sporozoan species and only one protein band with a similar electrophoretic mobility was found in the three gregarine species (52 Kd protein). Two G. garnhami subpopulations living in Locusta migratoria and Schistocerca gregaria exhibited the same ghost protein pattern. Thus, large differences were observed between species and not within species, and the protein electrophoretic analysis appears to be a powerful tool for taxonomic investigations in gregarines. Gregarina blaberae and G. garnhami glycoconjugates were compared after periodate/Schiff staining of the polyacrylamide gel slabs. Several glycoconjugates were reported to belong to the cytoplasmic fraction; and, in view of cytochemical and ultrastructural data, a contribution of these glycocomponents to the secretion of a mucus is discussed.

Notes: . Phytomonas davidi is found in the latex of species of the plant family Euphorbiaceae, including Chamaesyce hirta and C. hyssopifolia, to latitude 32°N in the southeastern United States. The hemipteran Pachybrachius bilobata scutellatus (family Lygaeidae) serves as an agent of transmission from plant to plant. Heavy infections of flagellates in the salivary glands were observed; trypanosomatids were less than one-half the size seen in the plants and in the gut of the bug.

Notes: . Flagellar cysts of Blastocrithidia triatomae form from active flagellates by diminution in size. The pellicular microtubules disappear. The inner layer of the cell membrane thickens progressively as the organism shrinks. The fully formed cyst has an electrondense layer that corresponds to the outer layer of the unit membrane. An electron-lucent layer is approximately twice the thickness of the middle layer of the unit membrane. Inside that is a 92 nm layer that may represent the cytoplasm. The nuclear content is in the form of whorled bundles of 10–15 nm fibrils. The kinetoplast was not seen in electron micrographs of cysts.

Notes: A study of plankton samples collected in the Western Sargasso Sea during nine cruises has shown discontinuity between samples from different latitudes as regards their percentage, abundance, and number of species of Acantharia. Limits separating zones rich in Acantharia from those poor in Acantharia were singled out. The latitude at which the difference in abundance occurs can vary. This is in accord with differences in primary production, phytoplankton, and mesopelagic fishes noted by other workers who believe that the points of these faunal changes correspond to a temperature change indicated as “thermal fronts.”

Notes: A simple and rapid method is described for the isolation of nuclei from the Florida red tide dinoflagellate Gymnodinium breve. The nuclei are free of cytoplasmic contamination and are active in endogenous RNA synthesis. The ratio of DNA: RNA: acidsoluble protein: acid-insoluble protein is 1:0.39:0.13:0.63, respectively, and each nucleus contains ca. 113 picograms of DNA. Electrophoretic analysis of the acid-soluble proteins reveals the presence of two histone-like proteins with molecular weights of 12,000 and 13,000.

Notes: Proteins of surface membranes and surface-related cytoskeletons in Tetrahymena vorax microstomes and macrostomes were compared by one-dimensional SDS polyacrylamide gel electrophoresis to see if protein differences could be detected that correlate with the transformation from one phenotype to the other. Some differences were observed. However, these alterations appear to result from the heat-shock procedure used to synchronize the microstome-to-macrostome transition. The apparent lack of transformationspecific changes in cortical proteins is discussed. Similarities and differences between cytoskeletal proteins of T. pyriformis GL-C and T. vorax are also noted and discussed.

Notes: The growth inhibition of Tetrahymena furgasoni (once known as “T. pyriformis W”) by C19 and C21 steroids of similar structure was measured by determining cell population at 24 h and 48 h following addition of the steroid. A cis-fusion of the A/B rings junction, unsaturation at C-1,2, or C-4,5 and carbonyl substitution all enhanced inhibition, whereas the presence of two hydroxyl groups decreased inhibition. The results indicated that the transformation of C19 and C21 steroids by this protozoon may be part of a detoxication mechanism.

Notes: The metabolism of phospholipids in synchronous Plasmodium falciparum-infected erythrocytes was studied over one cycle of 48 h by the incorporation of labeled palmitate, serine, choline, and myo-inositol into cellular lipids. The rates of incorporation of palmitate and serine into total phospholipids and of choline into phosphatidylcholine (PC) were linear with the maturation of the parasite, increasing by a factor of 2–5.6 according to the precursors. The rate of inositol incorporation into phosphatidylinositol was 9.6 times higher at the schizont stage than at the ring stage, with a marked increase in the second half of the cycle. A significant incorporation of palmitate into triglycerides also occurred during the schizont stage of the parasite. The incorporations of serine and palmitate into phosphatidylethanolamine (PE) and PC showed a net increase at approximately the twentieth hour of the cycle, while the radioactivities recovered in phosphatidylserine (PS) had already reached a maximum by this time. These findings indicate an instantaneous transformation of PS into PE and PC through a decarboxylation of PS into PE, then a methylation of PE into PC during the second half of the cycle. Although PS is a minor component of the Plasmodium parasite, our findings demonstrate the important role of this phospholipid as a precursor of PE and PC, which are major constituents of parasite phospholipids.

Notes: With the aid of energy-dispersive X-ray microanalysis, several protozoa were tested for content of cations within inorganic minerals. The skeleton of acantharia consists mainly of Sr with small quantities of Ca and Ba. Two Loxodes species contain nothing but Ba, while in some Remanella species Sr with small quantities of Ba were present. In one Geleia species, Ca with small quantities of Sr was found; in two Trachelocerca species from Sylt (Germany), Ba is there in addition. Another Trachelocerca species from northern Italy lacked Ba, but did possess Mn. In Prorodon only Ca was found.

Notes: Thin-sectioning and freeze-etching electron microscopy were applied to explore the structure and the temperature- and Ca2+-response of the different host and parasite membranes during intraerythrocytic development of Plasmodium knowlesi in Macacca mulatta. The plasma membrane of uninfected erythrocytes is temperature- and Ca2+-responsive: chilling to 4°C and exposure to 5 mM Ca2+ induces a slight decrease in IMP-frequency and the emergence of small IMP-devoid patches on P-faces. On parasite infection, the erythrocyte membrane becomes modified as indicated by an enhanced temperature-response and the appearance of caveolae, ca. 70–90 nm in diameter. The frequency of these caveolae is increased in schizont-infected erythrocytes. Moreover, electron dense plaques, ca. 40 nm in width, appear just beneath the erythrocyte membrane in late trophozoites and schizonts, thus indicating a further modification of the host cell membrane during parasite development. The membrane of the parasitophorous vacuole, derived from the host plasma membrane, dramatically reduces the IMP-frequency especially on the P-face upon parasite infection. This leads to an apparent reversal of the IMP-distribution persisting throughout the whole infection cycle. The parasite plasma membrane forms local compaction domains with the nuclear envelope in ca. 30% of the ring-stages and trophozoites, which disappear in late trophozoites and schizonts. Moreover, the IMP-frequency on plasma membrane fracture faces almost doubles during parasite development. Chilling induces a decrease in the IMP-frequency on P-faces of the plasma membrane. Surprisingly, however, the parasite plasma membrane and the vacuolar membrane respond to externally applied Ca2+ with almost a doubling of the IMP-frequency. The different parasite endomembranes also undergo characteristic changes during parasite development.

Notes: The leech Calliobdella vivida (Verrill) is the vector of Trypanoplasma bullocki. At 10°C, infective-stage flagellates were first present in the leech's proboscis sheath five days after feeding. At 5°C, infective-stage flagellates were not present in the leech's proboscis sheath until 10 days after feeding, but at 20°C, flagellates were located there as early as 24 h after feeding. Infected leeches retained flagellates through three subsequent feeds on uninfected fish. When flagellates were first observed in hogchoker, Trinectes maculatus (Bloch & Schneider), they were much larger than infective stages from the leech. Average flagellate length then decreased during early acute phase, but gradually increased thereafter. Peak parasitemia was greater in a hogchoker inoculated by only one leech but held at colder temperature than in a hogchoker inoculated by 45 leeches, suggesting that temperature may be more important than inoculum in determining peak parasitemia. Cell division in the fish host is described. SEM studies of fish blood flagellates revealed a pre-oral ridge and a cytostome.

Notes: Histones were prepared from chromatin of the eukaryotic (endosymbiont) nucleus of Peridinium balticum (Levander) Lemmermann. The amino acid composition of whole histone was rich in lysine and similar to that of Olisthodiscus luteus and Euglena gracilis. Electropheretic analysis of these proteins in acidic-urea disc gels revealed four major bands: one with a mobility slightly lower than that of calf thymus HI; and three others which comigrated with calf H2B, H2A, and H4, respectively. The low mobility band was soluble in 5% perchloric acid and was sensitive to FeCl3 destaining. Electrophoresis in slab gels containing 0.1% SDS revealed five major components, with approximate molecular weights of 23,000, 20,000, 15,000, 13,000, and 11,000, respectively. The 15,000 and 11,000 dalton histones had mobilities identical to those of calf H3 and H4, respectively. The two highest molecular weight components were soluble in 5% perchloric acid. No bands were found to comigrate with calf H2A or H2B but a band was present that migrated to a position intermediate between calf H2A and H4 (13,000 dalton histone). Two-dimensional gels consisting of acidic-urea gels in the first dimension and SDS gels in the second dimension revealed that the 20,000 dalton component and the 13,000 dalton component are not resolved in the acidic-urea gel. As a working hypothesis, it is suggested that two of the five bands seen in SDS gels represent an H1-like doublet, and two are analagous to H3 and H4, respectively. The remaining histone may replace H2A and H2B.

Notes: The iodinating reagent 1,3,4,6,-tetrachloro-3α,6α-diphenylglycoluril (IODOGEN3) was used to label antigens on zygotes of Plasmodium gallinaceum with parallel studies using lactoperoxidase-catalyzed radioiodination for comparison. Proteins labeled by the IODOGEN method are most probably on the surface of the zygote, as the pattern of labeled proteins analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis was very similar to the pattern of lactoperoxidase-labeled proteins. Furthermore, the labeled proteins represented only a subset of the total Coomassie Blue-stained proteins. The radioiodinated zygote proteins were immuno-reactive after IODOGEN or lactoperoxidase labeling. The IODOGEN method is technically much more simple than the lactoperoxidase method and does not require the addition of extraneous proteins or H2O2. The advantages of IODOGEN labeling, together with the essential equivalence of results obtained by these two, methods, make the IODOGEN method attractive for labeling parasite antigens in general.

Notes: A new method is described for mass cultivation of Euplotes aediculatus, a hypotrich ciliate containing omikron-symbionts. The ciliate cultures, continuously aerated in Erlenmeyer flasks (5000 ml) with 4500 ml medium, yield densities of 2300 cells/ml which are four to five times higher than cell densities of cultures grown in unaerated Fernbach flasks. Harvesting such cultures involves the application of 25-μm mesh sieves. Cells so concentrated can be purified by using columns or special chambers in which Euplotes migrates towards the cathodes in an electric field (field strength 7 V/cm).

Notes: Stocks of Entamoeba histolytica grown in a monoxenic culture system from the feces of nonhuman primates are compared with the eleven zymodemes of E. histolytica so far demonstrated from man. In a similar fashion, Entamoeba chattoni has also been grown and identified. Both E. histolytica and E. chattoni have been demonstrated in keepers of the primate collections. Comparisons have been made using the electrophoretic patterns of three enzymes: glucosephosphate isomerase [(GPI) E.C.5.3.1.9], phosphoglucomutase [(PGM) E.C.2.7.5.1], and L-malate—NADP+ oxidoreductase (oxaloacetate-decarboxylating) [(ME) E.C. 1.1.1.40]. Enzyme patterns of E. histolytica from the apes were found to be identical with three of those already demonstrated from man. The enzyme pattern of E. chattoni was distinctly different from that of any of the E. histolytica zymodemes. Other protozoa found in the single fecal sample examined from each subject are also listed.

Notes: SYNOPSIS. Dipodomys merriami Mearns and Dipodomys ordi Woodhouse were surveyed for coccidia in El Paso County, Texas. Infections with Eimeria chobotari. Eimeria dipodomysis and Eimeria balphae were 24.8%. 4.4%, and 11%, respectively, for D. ordi. Dipodomys merriami had an infection level of 23.8% with E. chobotari. Four animals concurrently harbored E. chobotari and E. balphae or E. dipodomysis or a new species Eimeria chihuahuaensis. Male and female host infection levels were not significantly different. The new species is described and photographs of 3 previously described Eimeria from Dipodomys are presented.

Notes: SYNOPSIS. Experimentally induced precystic stages and mature cysts from 3 clones of Tetrahymena rostrata were examined by light and electron microscopy. It was demonstrated by cytochemical staining and fine-structural observations that precystic stages release mucocyst material that provides for the production of a cyst wall. Early and late cysts also contain numerous autophagous vacuoles. In late cysts there is a replacement of depleted mucocyst organelles. The developmental evidence obtained from sampling of sequential developmental stages suggests an ∼24-h timetable of cytoplasmic events associated with encystment in this organism.

Notes: SYNOPSIS. Filamentous cyanobacteria are ingested through the cytopharynx of the ciliate Pseudomicrothorax dubius. The cytopharynx is a complex of microtubules and microfilaments located in a highly vesiculated cytoplasm, the phagoplasm. Two types of membrane-bounded phagoplasmic vesicles can be distinguished by their differences in size, fine structure, and acid phosphatase (AcPase) content. One type has a homogeneous, electron-dense interior which is AcPase-positive. These vesicles are present in fed cells and in unfed cells devoid of food vacuoles, and thus appear to be primary lysosomes. During phagocytosis, exocytosis within the cytopharynx of the primary lysosomes results in the elaboration of a food vacuole. The vacuole grows by incorporation of lysosomal membrane; lysosomal hydrolases are liberated into the vacuole. Within less than 1 second of AcPase's entry into the food vacuole, it is detectable within the cyanobacterial cytoplasm, and within 5 seconds, destruction of the cyanobacterial filament is observed. It is hypothesized that the rapidity of hydrolase penetration of the cyanobacterial cell wall is the result of the action of molecules analogous to the “killing agents” of neutrophil leukocytes, which rapidly render bacterial envelopes permeable. AcPase, and presumably other hydrolases, are present in the cyanobacterial filament when filament destruction occurs; they thus appear implicated in this process. Hydrolases may activate an autodestruction mechanism in the cyanobacterium. Firm adherence of the food vacuole membrane to the cyanobacterial filament is demonstrated, and its role in phagocytosis is discussed.

Notes: SYNOPSIS. The surface membrane potentials of suctorian genus Heliophrya were studied with intracellular electrodes. Resting membrane potentials averaged -32 mV, and spontaneous depolarizing potentials occurring at apparently random intervals and having a variety of waveforms were routinely observed. Such spontaneous potentials were correlated in time with visually monitored contractile vacuole activity. Individual contractile vacuoles had unique, although somewhat variable, electrical signatures. In the presence of an intracellular electrode all vacuoles contracted independently, but at approximately the same frequency. The amplitude of the electrical potentials increased when the membrane was hyperpolarized and decreased when it was depolarized. The sign of such potentials reversed at between -10 mV and the zero membrane potential. A 20% decrease in the membrane resistance was measured at the peak of the spontaneous depolarizing potentials.

Notes: SYNOPSIS. Female golden hamsters, either in the last week of pregnancy or in the first weeks of nursing, excreted in their feces variable numbers of pseudocysts of Tritrichomonas muris. Pseudocysts examined by electron microscopy had internalization of the 3 anterior flagella and the undulating membrane with its recurrent flagellum. The undulating membrane and the associated marginal lamellae were characteristic of T. muris. Pseudocysts gradually become motile after 2 or more hours of incubation in medium. The “excysted” trophozoites were identified ultrastructurally as T. muris. Newborn hamsters were not infected with T. muris at 3 days of age, but by the 7th day essentially all were found to have infected ceca, concomitant with cecal enlargement and the appearance of adult-type feces.

Notes: SYNOPSIS. Mating-type dependent cell pairing in Oxytricha hymenostoma was studied by both light autoradiography and transmission electron microscopy. The process results in the macronuclear bodies of the 2 partners fusing completely with each other 2 by 2. In each fusion the 2 chromatin masses, easily distinguishable from each other for the first 6 h of the pairing, progressively intermingled and underwent a sort of rearrangement so that they eventually acquired a uniform pattern. The possible mechanisms regulating macronuclear fusion and DNA amount in the daughter cells are discussed.

Notes: SYNOPSIS. Trypanosoma (Schizotrypanum) cruzi clones were derived from isolates of an acute human case of Chagas' disease (strain Esmereldo), a human case of T. cruzi infection (strain CAN-III) and from a naturally infected opossum (strain WA-250). The isoenzyme patterns and growth rates of the clones were stable during long-term cultivation, by serial passages, of the parasites in liquid medium. Both clones of strain Esmereldo were zymodeme II; the 2 clones of strain CAN-III, zymodeme III; and the 5 clones of strain WA-250, zymodeme I. The range in doubling times of the parasite populations in liquid medium were 36–49. 7 h (strain WA-250), 117.2–133.7 h (Esmereldo clones) and 169–208 h (CAN-III clones).

Notes: SYNOPSIS. Under aerobic conditions, we have determined glycerol uptake in the long slender (LS) bloodstream form of Trypanosoma (Trypanozoon) brucei brucei by studying glycerophosphate accumulation in the parasites. The coupled enzyme theory applies to the permeation-phosphorylation sequence. Glycerol passage through the plasma membrane is asymmetric, the efflux process being favored over the influx process. No free diffusion of glycerol can be detected even under conditions under which free glycerol accumulates within the cells; most probably, glycerol permeation is mediated by a specific transport system. In the absence of respiratory activities, glycerol is known to be an end-product of T. brucei glycolysis; its production from glycerophosphate should allow ATP synthesis. The observed efflux of free glycerol following intracellular accumulation of glycerophosphate confirms the hypothesis that glycerol production occurs through reversal of glycerol kinase activity.We conclude that in vivo the role of the carrier-mediated asymmetric permeation process is to prevent inhibition of the reversal of the glycerol kinase-mediated reaction by removing free glycerol.

Notes: SYNOPSIS. the following species are described from Hawaiian birds: Isospora brayi sp. n., with oocysts 27 × 26 μm and sporocysts 19 × 12 μm, from the Japanese white-eye. Zosterops japonicus Temminck & Schlegel; Isospora cardinalis sp. n., with oocysts 24 × 23 μm, and sporocysts 16 × 10 μm, from the cardinal, Cardinalis cardinalis (Linnaeus); Isospora ivensae sp. n., with oocysts 26 × 25 μm, and sporocysts 18 × 12 μm, from the spotted or white-throated munia, Lonchura punctulata (Linnaeus); Isospora loxopis sp. n., with oocysts 26 × 23 μm, and sporocysts 16 × 13 μm, from the amakihi or honeycreeper, Loxops virens (Gmelin); and Isospora phaeornis sp. n., with oocysts 27 × 19 μm, and sporocysts 16 × 11 μm, from the omao or Hawaiian thrush, Phaeornis obscurus (Gmelin). All the host birds belong to the order Passerorida.

Notes: SYNOPSIS. Equine protozoal myeloencephalitis (EPM) was diagnosed in 10 horses. By electron microscopy, schizonts were found in intact host cells of the spinal cords or, more frequently, free in the extracellular spaces. Developmental stages of schizonts differed morphologically, and the late stage of schizogony was characterized by endopolygeny. These findings permitted tentative identification of the protozoon as a Sarcocystis sp. Free merozoites were present in the extracellular spaces or in cells of the spinal cord. Pericytes of capillaries were most frequently parasitized by merozoites, but the cytoplasm of neurons, macrophages, intravascular and tissue neutrophils, and axons of myelinated nerve fibers also contained these organisms. the presence of parasites in the cytoplasm of tissue and circulating neutrophils suggests that this putative Sarcocystis sp. may have a hematogenous phase of infection.

Notes: SYNOPSIS. 20-Methylcholanthrene induced cyst formation in Nyctotheroides puytoraci when injected into its host Bufo regularis. Presumably this hydrocarbon or its metabolites reaches the parasites in the recta of treated host animals and induces encystment. However, injection of B. regularis with 0.5 mg of 20-methylcholanthrene + vitamin A palmitate (5,000 IU) inhibited the hydrocarbon-induced encystment of the parasites.

Notes: SYNOPSIS. Simple media for Tetrahymena, using rat gut or soybean as the nutrient source are described. Cultures can be maintained in these media up to one year at room temperature.

Notes: SYNOPSIS. A mutant of Tetrahymena thermophila. “pig,” excretes melanin precursors into the culture medium where spontaneous polymerization to melanin occurs. the precursors, probably oxidation products of catecholamines. are produced in large amounts by the mutant by decarboxylation of tyrosine and hydroxylation of the resulting tyramine. Overproduction and excretion of precursors by the mutant appears to result from elevated specific activity of L-aromatic amino acid decarboxylase (DOPA decarboxylase) (E.C. 4.1.1.26).

Notes: SYNOPSIS. Paramecium multimicronucleatum has been cultured for 20 years on a medium of salts, vitamins, amino acids, fatty acids, ribosides, and stigmasterol plus a little nondialyzable fraction (NDF) of baker's yeast. Fractionations of NDF identified 2 essentials: (a) in a fraction 〈 100,000 daltons which contained much protein and replaceable by ovalbumin and (b) in a fraction of 〈 300,000 daltons; this fraction contained much polysaccharide, replaceable by glycogen, which is 〉 300,000 daltons. For 2 years now P. multimicronucleatum has grown well with ovalbumin and glycogen replacing NDF. Besides ovalbumin, concanavalin A satisfies the protein requirement; this lectin attaches to sugar residues in glycogen. Studies with a fluorescent dye, PGA-1A, a stilbene derivative, provides further evidence for the polysaccharide requirement. This dye attaches to polysaccharides; when added to glycogen, and this in turn is added to a culture containing ovalbumin, fluorescent blue vacuoles appear within 2–3 h. When dye + glycogen were added to a culture without ovalbumin, no fluorescent vacuoles were found. A protein appears involved in formation of food vacuoles; this fits the pattern for endocytosis described in recent reviews. Besides glycogen, mannan gave good growth. Dextrin and amylopectin gave only fair growth through 7 serial transfers; glucose, maltose and amylose did not sustain growth. Strain 51 of P. tetratrelia, which grows well in NDF medium, grows well when NDF is replaced with ovalbumin and glycogen.

Notes: Oocysts of Eimeria caprovina sp. n. from the domestic goat, Capra hircus, are ellipsoidal, subspherical or slightly ovoid, usually flattened at the micropylar end. They measure 29.7 (26-36) × 23.7 (23-28) μ. the sporocysts are elongate ovoids, measuring 14.3 (13-17) × 8.3 (8-9) μ. with Stieda bodies at the narrow ends. the oocyst wait is 1.6 μ thick, smooth, dark-brown to brownish-yellow, and 2-layered. A micropyle. 6.2 (4-10) μ in diameter, polar granule, and sporocyst residuum are present: micropylar cap and oocyst residuum are absent.

Notes: Infective trypanosomes developed when Trypanosoma brucei was cultivated at 28 C in a liquid medium containing tsetse fly head-salivary gland explants. They were separated from the noninfective culture forms using DEAE-cellulose column chromatog-raphy. It was demonstrated by light and electron microscopy that the separated organisms were morphologically similar to metacyclic stages found in a tsetse fly and that they had a characteristic surface coat. Single metacyclic trypanosomes isolated from the cultures gave rise to infections in mice.

Notes: Sporozoites of Plasmodium berghei and Plasmodium knowlesi, incubated in normal serum readily interact with peritoneal macrophages of mice or rhesus monkeys, respectively. Interiorization of the sporozoite requires that both serum and macrophages be obtained from an animal susceptible to infection by the malaria parasite. Serum requirements for sporozoite attachment to the macrophage are less specific. Phagocytosis is not essential for the parasites to become intracellular. Our findings indicate that active penetration of the sporozoites into the macrophages does occur.

Notes: The uptake of [57Co]B13 (cyanocobalamin) by Euglena gracilis strain Z (ATCC 12716) occurred in 2 distinct phases-an initial rapid phase followed by a slower secondary phase. This secondary phase appeared after the saturation of the binding sites involved in the initial rapid phase and was energy-dependent and completely inhibited by 2,4-dinitrophenot, KCN and sodium azide. the subcellular localization of labeled cyanocobalamin taken up by the cell was mostly contained in the chloroplast fraction. the time course and the saturation kinetics of B12 uptake by purified chloroplast fraction indicated that this fraction and the intact cell had a similar affinity for the vitamin B12. This suggested that the chloroplasts contained the binding sites for vitamin 12 and might regulate the uptake process in the intact cell. the kinetic properties of the overall 12 uptake mechanism suggested that the initial phase represent the binding of vitamin 12 to the available sites on the chloroplast. the secondary phase may represent the de novo synthesis of new binding sites.

Notes: SYNOPSIS Amebae isolated from sediments of the Atlantic Ocean and Gulf of Mexico were maintained in continuous culture and most were identified to genus and species. Twenty-six species representing 12 genera were recognized from existing literature and several others (Flabellula, Mastigamoeba, Cochliopododium) were identified only to genus. One ameboflagellate and several small limax-type amebae which require further study also were isolated. Other sarcodmids belonging to the Heliozoida, Testocida, Leptomyxida, and Proteomyxida were identified only tentatively. the distribution of the amebae and ameba-like organisms was tabulated for the following geographic areas: Atlantic Ocean near Long Island, New York: Atlantic Ocean 16-65 miles offshore from New York and New Jersey: Atlantic Ocean 1-50 miles offshore from Maryland and Delaware: and the Gulf of Mexico 3.5-41 miles offshore from the southeastern United States. Amebae present in shellfish holding trays at Lewis. Delaware, were isolated, and identified to compare the distribution of species in laboratory tanks with those present in natural ocean bottoms. Published accounts of each collection site were reviewed to obtain specific data on contamination with sewage wastes, acid wastes, dredge spoils, and petroleum hydrocarbons. Two previously undescribed amebae were found to represent new genera and species and are described herein, one from the Delaware mariculture facility, and the other from the digestive tract of the blue crab, Callinectes sapidus, and the gill surface of the lady crab, Ovalipes ocellatus. Sarcodinids present in clean or stressed environments were listed, and genera and species that were widespread or apparently geographically restricted were recorded.