ALBERT

Notes: Trimyema compressum is a species included in the family Trimyemidae with the single genus Trimyema. This species has 50–60 somatic kinetics and three rows of kinetosomes surrounding the oral cavity. Two isolated groups of kinetosomes can also be observed on the right side of the oral region. The morphogenesis of bipartition is telokinetal; all the new infraciliary structures of the opisthe come from the longitudinal and postero-anterior proliferation of the last kinetosome of all the somatic kinetics. In the proter there is a reorganization of the oral infraciliature.As a result of our observations, we suggest that the systematic position of the genus Trimyema be changed from the subclass Vestibulifera to the subclass Gymnostomata. We also consider that this genus must be included in the suborder Trimyemina Jankowski, 1980.

Notes: Specimens of Pelomyxa palustris from five collecting sites had numerous nonmotile flagella. The structures are called flagella because of morphological similarities to flagella and because P. palustris has affinities with amoeboid flagellates. Flagella were photographed on living cells and studied by transmission and scanning electron microscopy. From 64 to 742 flagella per cell were estimated from scanning electron microscopy of ten cells 204 to 1269 μm in length. The nonmotile flagella arise from basal granules which were, in one strain, surrounded by radiating electron-dense microtubules. This strain also had excess axonemal microtubules. Abundant cytoplasmic microtubules were arranged in several different patterns. In about half of the P. palustris cells in which nuclei were studied, microtubules were either apposed to the nuclear membrane in a parallel alignment (with some also radiating) or radiating from the nuclear membrane (with none parallel). Bacteria associated with nuclei were of three characteristic types: Gram-negative rods, Gram-positive rods, and large rods. All nuclei within a given trophozoite had similar perinuclear features. Recent proposals for separation of Pelomyxa to its own phylum (based on its proposed primitive, unique nature) can not be justified. Pelomyxa is a complex, highly specialized organism adapted to live in a specific fresh-water environment. Mastigamoebid amoeboid flagellates of the genera Mastigamoeba, Mastigella, Mastigina, and possibly Dinamoeba are placed with Pelomyxa within the order Pelobiontida Page, 1976, emend., containing two families. Pelomyxidae Schulze, 1877, and Mastigamoebidae Goldschmidt, 1907.

Notes: SYNOPSIS. Thymidylate synthetase (E.C.2.1.1.45) has been demonstrated in unsporulated oocysts of Eimeria tenella. The properties of this enzyme have also been investigated in Tetrahymena pyriformis, as a protozoan model, and 7-day-old chick embryo, as a host model. The enzymes from E. tenella and chick embryo were inhibited by all concentrations of MnCl2 and MgCl2 tested. Tetrahymena pyriformis thymidylate synthetase was stimulated by low concentrations of both these cations but was inhibited by high concentrations. Subsequent data refer to chick embryo, E. tenella and T. pyriformis respectively: the apparent Km was 5.89 μM, 5.94 μM, and 0.53 M for the substrate dUMP: and 5.13 μM, 1.10 μM and 4.65 μM, respectively for the cofactor N5N10-methylenetetrahydrofolate. The pH optimum for the enzyme from both chick embryo and T. pyriformis was 8.0, with Tris-HCl buffer; activity of E. tenella thymidylate synthetase was still increasing at pH 8.2. The E. tenella enzyme was found to have a molecular weight of 4.6–4.9 × 105 daltons. The effects of nucleotides, inhibitors, and the omission of assay components on each enzyme are presented. Thymidylate synthetase from E. tenella is not greatly different from that of chick embryo, but does not resemble the enzyme from T. pyriformis. A case for using thymidylate synthetase as a chemotherapeutic target in the treatment of Eimeria infections remains. Indeed Eimeria may be considered as a model for infections caused by other protozoan parasites, such as Toxoplasma and Plasmodium, provided that suitable inhibitors can be found that are not toxic to the host.

Notes: SYNOPSIS. A protocol based on density differences between starved and fed cells and employing density gradient centrifugation has been devised to facilitate the isolation of auxotrophic mutants of cell lines derived from Tetrahymena thermophila strain B1868. First, a mass phenotype screening procedure was established whereby true auxotrophic mutants and slow-growing wild-type cells such as strain C* could readily be distinguished. Second, simulation experiments were performed in which wild-type cells starved first in non-nutritive buffer, then suspended in a defined medium lacking a single essential amino acid became significantly denser than the same cells when starved, then suspended in a complete defined medium. Finally, using the same protocol, a reconstruction experiment was carried out which resulted in effective separation of wild-type cells from cells of a tyrosine auxotroph. The overall procedure resulted in a 9-fold increase in the relative frequency of auxotrophic cells, while the density gradient centrifugation alone provided a 400-fold enrichment.

Notes: SYNOPSIS. Infectivity of Plasmodium gallinaceum (Brumpt) sporozoites isolated from midguts and salivary glands of experimentally infected Aedes fluviatilis (Lutz) was studied. The 2 populations were compared at 7, 8, and 9 days postisolation from mosquitoes, which were maintained at 27 C ± 1C and ∼75% relative humidity. Infectivity of the parasites was evaluated by the length of the prepatent period of the infection in 2-week-old chicks inoculated intramuscularly. Infection was caused by 7-day-old sporozoites from salivary glands, but not from midguts. Older sporozoites induced infection in all the inoculated chicks. The results suggested a somewhat higher infectivity of the 8- and 9-day salivary-gland parasites than of the oocyst sporozoites. However, unlike sporozoites from mammalian malaria, oocyst sporozoites from avian malaria were highly infective at this age.

Notes: SYNOPSIS. Sorogena stoianovitchae Bradbury & Olive, an epiphytic ciliate found in various parts of the world, has a trophic stage that feeds on members of the ciliate genus Colpoda. When grown in the presence of the food ciliate, it multiplies rapidly. When the cells become abundant they aggregate at the water surface on inserted plant fragments or floating pollen grains, the sides of culture dishes, or on floating films such as those deposited by bacteria or pollen grains. an aggregate mounds up and becomes ensheathed above the water level, after which the mass of cells called a sorogen rises aerially at the apex of a stalk deposited at its base. the tapering, noncellular stalk consists of a conspicuously furrowed sheath that encloses a mucilaginous matrix. At completion of stalk development the cells of the sorogen become encysted. the sorocysts are commonly discharged by fracturing of the drying sorus. Alternating light and dark conditions are required for sorocarp development.

Notes: SYNOPSIS. the antigenic types in populations of metacyclic trypanosomes of Trypanosoma brucei isolated from Glossina morsitans head-salivary gland trypanosome cultures and bloodstream forms in the early parasitemias produced from whole culture supernatant fluids containing metacyclic forms, were analyzed by the indirect fluorescent antibody test using clone-specific antisera. Metacyclic trypanosomes in cultures initiated with cloned bloodstream forms were heterogeneous with respect to their variable antigenic type (VAT). Trypanosomes comprising early parasitemias in immunosuppressed mice infected with metacyclics produced in cultures also had a range of VATs. Three of the VATs detected in the early parasitemias in mice have also been identified by other investigators in tsetse fly-transmitted populations of the same stock.

Notes: SYNOPSIS. A simple method is described for plating and cloning ciliates and other protozoa, based on a principle differing from that traditionally used for plating and cloning bacteria and other microorganisms. This procedure, referred to as the silicone-oil-plating-procedure (SOPP), involves vortexing small volumes of culture medium containing protozoa with larger volumes of a non-toxic silicone oil and plating the resulting unstable emulsion in small plastic petri plates. Discrete microdroplets of culture medium form containing protozoa entrapped and immobilized between the hydrophobic surfaces of the plastic petri dish and the oil. Protozoa, isolated by this method grow, divide, and multiply to form clones. the procedure may be used for plating and cloning protozoa in bacterized and axenic culture. Variations of the basic method may be applied to isolating protozoa from the wild, washing protozoa to remove microorganisms, screening for potential mutants, and for replica plating.

Notes: SYNOPSIS. Cyclic nucleotide phosphodiesterase [EC 3.1.4.17] was examined in Tetrahymena pyriformis strain NT-1. Enzymic activity was associated with the soluble and the particulate fractions, whereas most of the cyclic GMP phosphodiesterase activity was localized in the soluble fraction: the activities were optimal at pH 8.0–9.0. Although very low activities were detected in the absence of divalent cations, they were significantly increased by the addition of either Mg2+ or Mn2-. A kinetic analysis of the properties of the enzymes yielded 2 apparent KIII values ranging in concentration from 0.5 to 50 μM and from 0.1 to 62 μ M for cyclic AMP and GMP. respectively. A Ca2+-dependent activating factor for cyclic nucleotide phosphodiesterase was extracted from Tetrahymena cells, but this factor did not stimulate guanylate cyclase [EC 4.6.1.2] activity in this organism. On the other hand, Tetrahymena also contained a protein activator which stimulated guanylate cyclase in the presence of Ca2+, although this activator did not stimulate the phosphodiesterase. the results suggested that Tetrahymena might contain 2 types of Ca2+-dependent activators, one specific for phosphodiesterase and the other for guanylate cyclase.

Notes: SYNOPSIS. the cell size of Didinium nasutum was found to be dependent on the size of the Paramecium species available as prey. Didinium feeding on P. tetraurelia averaged 5.6 × 105μm3. the cell volume of Didinium increased with increasing prey size for the 5 prey species tested, to 9.1 × 105μm3 for Didinium feeding on P. caudatum. Didinium nearing a cell division ranged in size from 8.6 × 105μm3 on P. tetraurelia to 12.9 × 105μm3 on P. caudatum. the range in cell volume is such that Didinium feeding on P. caudatum are larger than the size at which Didinium divide when feeding on P. tetraurelia. This morphologic plasticity in cell volume allows Didinium to exploit a wide size range of Paramecium species as prey. It is proposed that the size of a Didinium may have profound effects on its ability to encounter and capture prey of different sizes.

Notes: RESUME. Chacun des 45–80 organelles adoraux de Bursaria truncatella O. F. Müller est constitué de 3 rangées de cinétosomes et l'aire buccale droite est couverte de nombreuses doubles rangées de cinétosomes. La stomatogenèse débute par la désorganisation et la résorption des organelles buccaux postérieurs. Puis, il y a désorganisation des rangées parorales de cinétosomes et multiplication des cinétosomes sur l'aire orale droite, en měme temps que sont rompues, selon une ligne oblique, un certain nombre de cinéties somatiques. La prolifération des cinétosomes aux extrémités des cinéties. de part et d'autre de la ligne de rupture, aboutit, d'une part, à la formation d'un champ anarchique qui est le primordium oral droit de l'opisthe, d'autre part, à la formation de nombreux doublets qui constituent chacun le primordium de chaque organelle adoral. Après la séparation des tomites, les cinétosomes de l'aire droite s'ordonnent en doubles rangées et les organelles adoraux se complètent par addition d'une 3ème rangée de cinétosomes. Les cinétosomes somatiques sont jumelés, reliés par 2 desmoses. Les fibres transverses postérieures et les fibres postciliaires forment de longs rubans de microtubules dirigés vers l'arrière et juxtaposés dans les crětes intercinétiennes. Les doubles rangées droites de cinétosomes buccaux sont assimilables à des stichodyades. Les organelles des cinétosomes adoraux portent des rideaux de fibres postciliaires convergents ou divergents. La rangée postérieure de chaque organelle est non ciliée. Par son type de stomatogenèse, par sa structure corticale, par l'ultrastructure des organelles adoraux, Bursaria appartient aux Colpodidea, ce qui suggère des remarques de plusieurs types.SYNOPSIS. In Bursaria truncatella O. F. Müller, each of the 45–80 adoral organelles is composed of 3 rows of kinetosomes, and the right buccal area is covered by many double rows of kinetosomes. Stomatogenesis begins by disorganization and disappearance of the posterior buccal organelles. Next, there is disorganization of the paroral rows of kinetosomes and multiplication of kinetosomes in the right oral area; at the same time, some somatic kineties are disrupted along an oblique line. Multiplication of kinetosomes at the extremities of the kineties, on both sides of the disruption, leads to the formation of an anarchic field which is the right oral primordium of the opisthe and the formation of doublets each of which constitutes an adoral organelle. After the separation of the tomites. the kinetosomes in the right buccal area position themselves, and the adoral organelles are completed by the addition of a 3rd row of kinetosomes. Somatic kineties are formed by successive pairs of ciliated kinetosomes united by 2 desmoses. the long posterior transverse ribbons and the postciliary ribbons extend posteriad, overlapping in the pellicular ridges. Oral rows of kinetosomes on the right can be compared with stichodyads. the adoral kinetosomes have convergent or divergent postciliary ribbons. the posterior row of kinetosomes in each organelle is not ciliated. By the type of stomatogenesis, the cortical ultrastructure, the ultrastructure adoral of its organelles, Bursaria belongs to the Colpodidea.

Notes: SYNOPSIS Free-living marine ciliates occur in the interstitial spaces of a wide vareity of filamentous and particulate substrata, on the surfaces of planar substrata, and in the plankton. In addition, they are found in association with a wide variety of plant and animal hosts. In this paper I review the progress during the past decade in understanding the distribution of marine ciliates, with particular emphasis on the relationship between ciliate biogeography and the species problem. It is concluded that as a general rule among marine ciliates, genera and species complexes are cosmopolitan. Specific locales may support a confusing array of sibling species or subspecific morphologic variants. Because the distributional processes and breeding biology of marine ciliates are only beginning to be understood, conventional ideas that marine ciliate species are cosmopolitan may require modification.

Notes: SYNOPSIS The subkingdom Protozoa now includes over 65,000 named species, of which over half are fossil and ∼ 10,000 are parasitic. Among living species, this includes ∼ 250 parasitic and 11,300 free-living sarcodines (of which ∼ 4,600 are foraminiferids); 1.800 parasitic and 5,100 free-living flagellates: ∼ 5,600 parasitic “Sporozoa” (including Apicomplexa, Microspora, Myxospora, and Aseetospora); and ∼ 2,500 parasitic and 4,700 free-living ciliates. There are undoubtedly thousands more still unmamed. Seven phyla of PROTOZOA are accepted in this classification—SARCOMASTIGOPHORA. LABYRINTHOMORPHA, APICOMPLEXA, MICROSPORA, ASCETOSPORA, MYXOSPORA, and CILIOPHORA. Diagnoses are given for these and for all higher taxa through suborders, and representative genera of each are named. the present scheme is a considerable revision of the Society's 1964 classification, which was prepared at a time when perhaps 48,000 species had been named. It has been necessitated by the acquisition of a great deal of new taxonomic information, much of it through electron microscopy. It is hoped that the present classification incorporates most of the major changes that will be made for some time. and that it will be used for many years by both protozoologists and non-protozoologists.

Notes: SYNOPSIS Leishmania donovani amastigote-to-promastigote transformation is inhibited by homogenates of infected hamster liver and spleen. This inhibitory activity is localized in the 100,000 g pellet fraction. Tests with lysates of adherent (macrophyages) and nonadherent (lymphocytes) spleen cells indicated that the inhibitory activity resided in the lymphocytes, specifically in the 100,000 g pellet fraction.

Notes: SYNOPSIS Antibodies induced in rabbits against Paramecium multimicronucleatum syngen 2 prevent sexually reactive cells from clumping, pairing, and forming cytoplasmic fusions. A biologic assay for the detection of these antibodies (designated blocking antibodies) is described. the blocking antibodies, unlike the immobilization antibodies, are produced against breis of sexually reactive cells and nonreactive cells of 2 types, nonstarved and immature. Isolated cilia from reactive cells of either mating type are weak immunogens for blocking antibodies. No correlation between the mating type specificity (III or IV) and these antibodies has been detected. Blocking antibodies can be absorbed with living cells, of which sexually reactive ones are the most effective absorbers, while immature ones are the least effective.

Notes: SYNOPSIS A method is described for the axenic mass cultivation of Paramecium tetraurelia strains 51s and 299s. the ciliate is grown in an enriched axenic medium developed by Soldo, Godoy & van Wagtendonk. Under continuous shaking on a rotary shaker, cultures were grown in one-liter Erlenmeyer flasks with 330 ml medium yield cell densities of 32,000 cell/ml and 20,000 cells/ml for strains 299s and 51s respectively. Doubling time is considerably shorter under these conditions than in the conventional static cultures. A 20-liter airlift bioreactor is described in detail which can be used successfully to otain up to 100 g wet weight of Paramecium in a single run; in this reactor the cell density reaches 38,000 cells/ml for strain 299s. and 23,000 cells/ml for 51s. This technic should facilitate the study of minor protein components of the ciliate.

Notes: SYNOPSIS The cadmium ion (Cd2+) was accumulated by Amoeba proteus in all cellular fractions, the highest level being associated with the cytosol fraction. On gel separation of the cytosol fraction, Cd-binding protein appeared in 2 peaks: one 〉45,000 MW (peak I) and the other 12,000 MW (peak II). Added cysteine increased the total Cd2+ taken up by the cells and resulted in disproportionate increase of Cd incorporated into the Cd-binding protein of peak II. the Cd-binding protein of peak II is analogous to the low-MW, Cdbinding proteins in Anacystis nidulans, Mytilus edulis, and to the metalloprotein of some vertebrates.

Notes: . Moles from Japan were examined for coccidian oocysts, and 67 of 77 (87%) hosts were infected including 8 of 11 (73%) Euroscaptor mizura, 31 of 36 (86%) Mogera kobeae, 17 of 17 M. tokudae, and 11 of 13 (85%) M. wogura. Of 67 infected hosts, 57 (85%) had multiple infections representing 2–5 coccidian species when examined. All oocysts in the infected fecal samples remained unsporulated and the absence of sporulation may be the result of storing feces from Japanese moles in 2% aqueous H2SO4. Five structurally distinct forms of unsporulated oocysts were found in E. mizura, and five distinct forms of unsporulated oocysts were also seen in Mogera spp. Two of the forms from E. mizura were similar to forms from Mogera spp., and the five forms from Mogera were shared freely between the three Mogera species. This is the first systematic survey of Japanese moles for coccidia.

Notes: . Total cellular DNA from the ciliates Halteria grandinella and Trithigmostoma cucullulus was analyzed by agarose gel electrophoresis. The macronuclear DNA (MAC DNA) of Halteria consisted of very small fragments, which suggests that the MAC DNA organization of oligotrichs resembles that of hypotrichs (gene-sized DNA). The MAC DNA of Trithigmostoma, a cyrtophorid having a heteromeric MAC, also existed as small fragments, but with a significant fraction (20–30%) comprising larger molecules unresolved by the method used. It is suggested that MAC heteromery is related to the differential localization of two kinds of DNA molecules of different sizes.

Notes: . Moles from England were examined for coccidian oocysts and all 64 Talpa europaea were infected; of 64 infected hosts, 56 (88%) had multiple infections representing two to six coccidian species when examined. Oocysts in 31 of the 64 samples remained unsporulated. Three eimerians and one isosporan were studied from the 33 fecal samples that had sporulated oocysts and these are described as new species; Cyclospora talpae Pellérdy & Tanyi, 1968, and Isospora sofiae (Golemansky, 1978) Levine & Ivens, 1979, are redescribed; and Cyclospora sp., similar to C. talpae, is discussed. Sporulated oocysts of C. talpae are ellipsoidal, 14.3 × 9.6 (12–19 × 6–13) μm with sporocysts ovoid, 9.4 × 5.7 (6–13 × 4–8) μm; it was found in 21 of the 33 (63.6%) sporulated samples. Sporulated oocysts of Cyclospora sp. are subspheroidal to ellipsoidal, 12.5 × 8.9 (10–14 × 6–12) μm with sporocysts ovoid, 8.6 × 5.3 (6–10 × 4–6) μm; it was found in 21 of the 33 (63.6%) sporulated samples. Sporulated oocysts of Eimeria avonensis n. sp. are elongate-ellipsoidal, 15.0 × 9.6 (13–20 × 7–12) μm with sporocysts ovoid, 6.6 × 3.6 (5–9 × 3–7) μm; it was found in 15 of the 33 (45.5%) sporulated samples. Sporulated oocysts of Eimeria berea n. sp. are subspheroidal, 12.1 × 10.5 (10–15 × 8–14) μm with sporocysts ovoid, 6.3 × 3.9 (5–10 × 2–5) μm; it was found in 8 of the 33 (24.2%) sporulated samples. Sporulated oocysts of Eimeria globula n. sp. are spheroidal, 20.9 × 19.9 (19–24 × 17–21) μm with sporocysts elongate-ovoid, 11.5 × 6.9 (9–16 × 6–9) μm; it was found in 3 of the 33 (9.1%) sporulated samples. Sporulated oocysts of Isospora sporopointaea n. sp. are subellipsoidal to ellipsoidal, 17.1 × 11.4 (13–21 × 8–14) μm with sporocysts ellipsoidal with both ends pointed, 11.9 × 5.9 (9–16 × 4–8) μm; it was found in 27 of the 33 (81.8%) sporulated samples. Sporulated oocysts of I. sofiae are spheroidal to subspheroidal, 12.2 × 11.0 (9–16 × 8–15) μm with sporocysts ovoid, 9.1 × 5.2 (6–13 × 3–8) μm; it was found in 25 of the 33 (75.8%) sporulated samples. To date, the coccidian parasites of talpids include two cyclosporans, 12 eimerians, and six isosporans, exclusive of the four new species described here.

Notes: . This report deals with a group of ciliated protozoa with short ciliary bands found mainly in the cecum of black rhinoceros, Diceros bicornis (Linnaeus, 1758), and white rhinoceros, Ceratotherium simum (Burchell, 1817) from southern Africa. A new genus, Rhinozeta, based on the sum total of the characteristics of seven new related species is described. The species described are R. rhinozeta n. sp., R. triciliata n. sp., R. caecalis n. sp., R. addoensis n. sp., R. cristata n. sp., R. multiplatus n. sp., and R. unilaminatus n. sp. The specific features of the new genus make it incompatible with any of the known families of the Order Entodiniomorphida containing the ciliates present in the digestive tract of herbivorous mammals. This merits the creation of a new family, the Rhinozetidae.

Notes: . Four isosporan species are described from the small tree finch, Camarhynchus parvulus from Isabela Island on the Galapagos Archipelago. Isospora exigua n. sp. oocysts subspheroidal, one-layered, smooth, yellow-brown color, 20.4 × 20.1 (20-23 × 18-23) μm, with no micropyle, residuum, or polar body. Sporocysts ovoidal, 14 × 9.5 (13-15 × 8-10) μm, with small Stieda and substieda bodies and irregular-shaped residuum. Isospora rotunda n. sp. oocysts subspheroidal, single-layered, smooth, yellow-brown wall with large polar body and no micropyle or residuum, 20.9 × 20.8 (20-24 × 19-23) μm. Sporocysts ovoidal, 15 × 9.7 (13-16 × 9-10) μm with knob-like Stieda bodies, prominent substieda bodies and round residuum. Isospora fragmenta n. sp. oocysts subspheroidal with no micropyle or residuum but with many splinter-like polar granules and a smooth, colorless, single-layered wall, 25.3 × 24.2 (24-27 × 23-25) μm. Sporocysts piriform 15.4 × 11.5 (14-17 × 11-12) μm with knob-like Stieda bodies, prominent substieda bodies, and irregular-shaped residuum. Isospora temeraria n. sp. oocysts ellipsoidal with one polar body, no micropyle or residuum, and wall of a single layer, smooth, yellow-brown color, 25.4 × 21.1 (21-30 × 17-23) μm. Sporocysts piriform, 15 × 10 (14-15 × 9-11) μm with knob-like Stieda bodies, prominent substieda bodies, and a round residuum. One woodpecker finch, Cactospiza pallida, was found to be infected with I. exigua, and a warbler finch, Certhidea olivacea was infected with I. fragmenta.

Notes: The ultrastructural features of cell division in the biflagellate, phagotrophic euglenoid, Entosiphon sulcatum, have been examined. Prophase is marked by the appearance of daughter feeding apparatuses and the emergence of two additional flagella. Pairs of flagella begin to migrate laterally along the surface of the elongating nucleus and remain lateral to the developing spindle poles. As the nucleolus elongates, it becomes dumbbell-shaped and the chromosomes move to the center of the nucleus, forming a loosely organized metaphase plate. Microtubules from opposing spindle poles attach to one of the pair of kinetochores found on each chromosome. The initial chromosome separation occurs during anaphase as the nucleus elongates. The length of the chromosomal microtubules does not decrease until late anaphase/early telophase. As the nucleus elongates, it forms a dumbbell-shaped structure. Most of the remaining microtubules are positioned in the interzone between the forming daughter nuclei. The interzonal spindle becomes somewhat constricted but remains intact until it is broken by the impinging cleavage furrow. Replication of the pellicular strips is not completed until late in cytokinesis.

Notes: The storage carbohydrate granules from Euglena and Pavlova were compared by light and electron microscopy. Freezeetch studies demonstrated that while both types of granules are crystalline, they have different structures. The elemental microfibril of the euglenoid granule measures 4 nm, and the elemental striation of the granule from Pavlova is 22 nm. The granules each have a unique X-ray diffraction pattern. The storage carbohydrate granules from Pavlova are not the same as paramyton, and the term “paramylon” should be reserved for the euglenoid storage carbohydrate.

Notes: A random-walk model of food-searching behavior is considered for the microzooplankton. It is suggested that in still waters a random walk of the conventional sort, modeled by a Wiener process, is less efficient than a Levy walk (a random walk whose excursions follow a Levy distribution) with Levy parameter less than two. For Levy parameter less than one, however, little advantage is gained by further reduction. In turbulent water, on the other hand, dispersion due to a random walk is dominated by the turbulent diffusion of the medium so that the Levy parameter appears to be less important. The effect of chemosensory responses is considered. It is suggested that these are most useful in still water, whereas in turbulent water their value would be less, and a non-specific filtering behavior might be more plausible.

Notes: Serotonin and catecholamines affect the regeneration of cilia in Tetrahymena thermophila in a dose-dependent manner: micromolar concentrations are stimulatory, whereas millimolar concentrations have little or no effect. This conclusion is based on motility measurements in regenerating cells and on ciliary counts in scanning electron micrographs. In addition, the recognition mechanism for each hormone appears to be specific and independent. Our results suggest an evolutionary link with hormonal mechanisms in multicellular eukaryotes.

Notes: Amicronucleate Paramecium tetraurelia fails to develop an oral apparatus but resorbs the pre-existing one in the sexual cycle. Previous cytological studies have revealed the presence of a shallow, oral depression with a few scattered basal bodies after autogamy or conjugation. The present ultrastructural study of the ectoplasm beneath the oral depression revealed only the presence of a disorganized filamentous reticulum and small blocks of cytopharyngeal ribbons. In addition, beneath the oral depression, a large number of discoidal vesicles were found, similar to but larger in diameter than those normally associated with the cytopharynx of vegetative cells. The oral structures found in amicronucleates in autogamy might be remnants of the pre-existing oral apparatus, but the possibility that they were newly developed was not excluded. The morphogenetic interest of these structures was discussed. It is evident that amicronucleates in autogamy do not develop an extensive oral ultrastructural organization.

Notes: The 115,000-molecular-weight antigen of Trichomonas vaginalis was characterized using monoclonal antibodies developed to three different strains of T. vaginalis and one strain of Tritrichomonas foetus. The antigen was found to be present on all strains or isolates of T. vaginalis examined and was demonstrated to be located on the external surface plasma membrane by agglutination assays and complement-mediated lysis assays. Characteristics of the antigen were assessed with a proteolytic enzyme and periodate oxidation. Periodate treatment of whole T. vaginalis abrogated binding for eight antibodies while use of pronase-treated antigen resulted in loss of antibody binding for two different antibodies. Screening of 19 axenized clinical isolates of T. vaginalis and one strain each of T. foetus and Giardia lamblia with type-specific antibodies delineated three major groups of T. vaginalis based on antigenic specificities (epitope distributions) within the 115,000-molecular-weight antigen. In addition, one epitope of the 115,000-molecular-weight antigen was found only on the immunizing strain. Two epitopes were present on all T. vaginalis isolates as well as T. foetus and G. lamblia. One epitope was common to all T. vaginalis except one. A minimum of six different epitopes of the 115,000-molecular-weight antigen were identified. Antigens purified with type-specific or “common” monoclonal antibodies shared the same partial peptide maps demonstrating relatedness.

Notes: Cells of Entamoeba histolytica grown over a period of four days contained NADP+-dependent alcohol dehydrogenase exclusively inside the cells. No activity of this enzyme could be found in the growth medium after harvesting the cells. Under the same conditions, acid phosphatase, β-N-acetylglucosaminidase, esterase, α-glucosidase, and different amylases of the parasite were found both inside the cells and in the medium. The activities present in the cell homogenate and in the medium before and after growth of the amoebas were partially separated by gel filtration on Sephadex G150 and G75, respectively. The comparison of the elution diagrams revealed that NADP+-dependent alcohol dehydrogenase, acid phosphatase, esterase, and amylases occurred as multiple forms inside the cells. These activities, as well as β-N-acetylglucosaminidase and α-glucosidase, were released into the extracellular environment to a different degree. The enzymes originating from the parasite were identified and distinguished from those of the ingredients of the growth medium according to their molecular mass and pH optimum. Furthermore, the amoebic origin of the secreted enzymes was shown on the basis of their inhibition by antibodies prepared against the supernatant fraction of the homogenate.

Notes: A previously reported microsporidium in Penaeus merguiensis de Man, 1888 (Decapoda, Penaeidae), in the Philippines has been identified as a species of Agmasoma Hazard & Oldacre, 1975, and named Agmasoma aquinoae n. sp. (Microsporida, Thelohaniidae).

Notes: The fine structure of trypanosomatids from the plants Euphorbia hyssopifolia, E. characias, E. pinea, and Manihot esculenta, cultivated under axenic conditions, was compared. All parasites had a structural organization characteristic of members of the Trypanosomatidae. The organization of the membranes of the endoplasmic reticulum (ER) varied according to the isolate. In Phytomonas francai (isolated from M. esculenta), the ER cisternae form parallel rows. In Phytomonas sp. from E. characias, a para-crystalline array of the ribosomes attached to the ER membranes was observed. The ER membranes found in Phytomonas sp. from E. pinea seemed to originate from the central portion of the protozoon, branching in all directions. The peroxisome-like organelle (glycosome) found in Phytomonas sp. from E. hyssopifolia and E. characias occasionally showed an organized array. Taken together, the morphological observations make it possible to distinguish the four isolates of Phytomonas.

Notes: Here we describe a method that allows the isolation of intact trypanosomatid symbionts in amounts sufficient for biochemical analysis. The isolated symbionts retain their characteristic morphological features and are reasonably free of subcellular debris. They actively incorporate [3H]leucine and [35S]methionine into proteins. Chloramphenicol and rifampicin at 50 μg/ml almost completely inhibit the incorporation of protein precursors. The inhibition of protein synthesis by the antibiotics provides direct evidence for the existence of a prokaryotic protein-synthesizing system in this unusual intracellular structure. The pattern of protein synthesis of the isolated symbionts is complex. Several symbiont polypeptides are absent or poorly represented in the flagellate.

Notes: Eimeria bovis has two types of first-generation merozoites with distinct ultrastructural characteristics. Type I merozoites were relatively large (X = 13.2 times 1.5 μm) and crescent-shaped, contained numerous micronemes and amylopectin granules, had a posteriorly located nucleus and a conical-shaped posterior tip, and were highly motile and capable of penetrating cultured cells. Type II merozoites were small (X = 5.9 times 0.9 μm) and spindle-shaped, had a centrally-located nucleus, few micronemes, few or no amylopectin granules, a dome-shaped posterior tip and little motility, and appeared to be incapable of penetrating cultured cells. It is possible that these two types of merozoites have considerably different roles in the life cycle of E. bovis.

Notes: To better understand the general distribution of glycoproteins and the distribution of specific glycoprotein-bound sugar residues in Paramecium, a survey of the binding pattern of selected lectins was carried out in P. tetraurelia, P. caudatum, and P. multimicronucleatum. Lectins studied were concanavalin A (Con A), Griffonia simplicifolia agglutinins I and II (GS I and GS II), wheat germ agglutinin (WGA), Ulex europaeus (UEA I), peanut agglutinin (PNA), Ricinis communis toxin (RCA60) and agglutinin (RCA120), soybean agglutinin (SBA), Bauhinia purpurea agglutinin (BPA), Dolichos biflorus agglutinin (DBA), and Maclura pomifera agglutinin (MPA). Those giving the most distinctive patterns were Con A, GS II, WGA, UEA I, and PNA. No significant differences were found between the three species. Concanavalin A, a mannose/glucose-binding lectin, diffusely labeled the cell surface and cytoplasm and, unexpectedly, the nuclear envelopes. Events of nuclear division, and nuclear size and number were thus revealed. Both WGA and GS II, which are N-acetylglucosamine-binding lectins, labeled trichocyst tips, the cell surface, and the oral region, revealing stages of stomatogenesis. The lectin WGA, in addition, labeled the compartments of the phagosome-lysosome system. The lectin PNA, an N-acetyl galactosamine/galactose-binding protein, was very specific for digestive vacuoles. Finally, UEA I, a fucose-binding lectin, brightly labeled trichocysts, both their tips and body outlines. We conclude that a judicious choice of lectins can be used to localize glycoproteins and specific sugar residues as well as to study certain events of nuclear division, cellular morphogenesis, trichocyst discharge, and events in the digestive cycle of Paramecium.

Notes: A simple method for isolation of plasma membrane from Acanthamoeba using self-generating gradients of Percoll is described. To obtain a membrane marker, intact amoebae were radioiodinated and the distribution of the radiolabel was followed through the plasma membrane isolation procedure. The purity of isolated plasma membrane was assessed by enrichment of radiolabel, by electron microscopy, and by enzymatic assays for contaminating membranes. As judged from enrichment of radiolabel, a 37-fold purification of plasma membrane was obtained. We estimate that 80% of the total protein was from plasma membrane and 10% from membrane-associated actin.

Notes: . We have studied linkage disequilibrium in natural populations of Trypanosoma cruzi, the agent of Chagas’ disease, by analyzing (i) a set of 524 stocks from the whole geographical range of the parasite, characterized at four gene loci coding for enzymes; (ii) a subsample of 121 stocks characterized at 12 enzyme loci; and (iii) a subset of 386 stocks from six locations in Bolivia, characterized by four enzyme loci. Our results show that the linkage disequilibrium reaches the maximum possible value, given the observed allelic frequencies, for almost all the locus pairs. This result is most consistent with the hypothesis that genetic recombination is absent or very rare in T. cruzi natural populations. Partition of the linkage disequilibrium variance for the six Bolivian populations shows that both inter- and intrapopulation components are substantial and that the relationships among the components are DIS2 〈 DST2, and D'IS2 〈 D'ST2. These inequalities are interpreted as the result of an interplay between genetic drift, rare or absent mating, and clonal selection in generating linkage disequilibrium in T. cruzi populations.

Notes: . The population of organelles in the periphery of the contractile vacuole of Amoeba proteus has been studied throughout the cycle using proper fixation and dehydration procedures on serial semithin and thin sections. Three distinct types of perivacuolar vesicles which arise successively from one another in the course of the cycle have been described. Size relationships have been determined by stereology.

Notes: . The binding of synthetic peptides modeled from the sequence of the cell attachment site of fibronectin to T. cruzi trypomastigote surface receptors was investigated by fluorescence-activated cell-sorting analysis using fluorescein-labeled peptides. Peptides with the sequence Arg-Gly-Asp-Ser bound to the parasite surface. A low percentage of fresh parasites recently liberated from infected fibroblasts had the capacity to bind the peptide. In contrast, these parasites showed a time-dependent several-fold increase in their ability to bind the Arg-Gly-Asp-Ser-containing peptides during extracellular incubation. From these observations, it appears that the expression of surface receptors on a particular, mature stage of the parasite parallels its ability to adhere to and infect host cells.

Notes: . Geographically diverse strains and clones of Tritrichomonas foetus have been examined with respect to their expression of a major surface antigen of approximately 150,000 relative molecular weight (Mr), designated the 150 Ag. Radioiodination and 13S-methionine labeling of T. foetus followed by immunoprecipitation with monoclonal antibodies (MAbs), separation of polypeptides by SDS-PAGE, and autoradiography or fluorography confirmed the parasite origin of the 150 Ag. The results of flow cytometry analysis employing a panel of MAbs against live T. foetus parasites revealed that from 5 to 84% of individuals in a given population of T. foetus expressed a particular epitope of the 150 Ag. All strains and clones were positive for surface expression of epitopes of this antigen. These results show that the 150 Ag is widely distributed in populations of T. foetus, confirm the surface location of this antigen, and suggest its importance as a target for protective immune responses.

Notes: . The hypotrich ciliate Onychodromus quadricornutus is remarkable in its potential for voluminous size (up to 900 μm in length), its possession of a unique set of four dorsal spines or horns, and its capability to express two kinds of developmental polymorphism induced by intraspecific predation. Cell length frequencies in replicates of a well-fed clone show normal distributions; starvation followed by intraspecific predation, however, induces cells within a clone to transform into two size classes: small lanceolate cells and cannibal giants. Induction experiments indicate that a substance released by cannibal giants stimulates defensive spine growth in clonemates within 24 h. Giants can also induce spine growth in non-clonemates. Furthermore, O. quadricornutus cells exposed to the predacious ciliate Lembadion magnum also develop hypertrophied spines. Selection experiments show that conspecific giants prey on cells with undeveloped spines (〈 20 μm in length) to a much greater extent than on cells with developed spines (〉40 μm in length). Transformation of a population of similarly sized O. quadricornutus cells into two different size classes may function to increase the range of potential prey sizes available to the O. quadricornutus population; hypertrophied spines appear to function as an inducible defense against intermittent predators appearing in the system including conspecific giants. This is the first reported case of a defensive developmental polymorphism induced by intraspecific predation.

Notes: . We studied the cellular regulation of vesicle exocytosis by Entamoeba histolytica utilizing release of endocytosed 125iodine (125I) labeled tyrosine conjugated dextran; 125I-dextran entered the acid pH vesicles of the amebae and was not degraded during these studies. Exocytosis was temperature dependent with 74%, 36%, 4%, and 0% of 125I-dextran released after 120 min at 37°C, 31°C, 25°C, and 4°C, respectively (P 〈 0.01 for each). Exocytosis at 37°C was inhibited by cytochalasin D (10 μg/ml), EDTA (10 mM), or the putative intracellular calcium antagonist TMB-8 (250 μM) (P 〈 0.01 for each at ≥ 60 min). Calcium ionophore A23187 (1 μM) enhanced exocytosis at 5 and 15 min (P 〈 0.01). Elevation of vesicle pH with NH4Cl (10 mM) had no effect on release of 125I-dextran; phorbol myristate acetate (10−6 M) increased exocytosis by 46% at 30 min (P 〈 0.01). Centrifugation of amebae with target Chinese hamster ovary cells resulted in decreased 125I-dextran release into the cell supernatant after 30 and 60 min at 37°C (by 40% and 42%, respectively, P 〈 0.01); release of 125I-dextran returned to control values with addition of 1.0 g% galactose or GalNac but not with mannose or N-acetyl-D-glucosamine. Amebic phagocytosis of serum-exposed latex beads had no effect on release of dextran by amebae (n = 16). Exocytosis of acid pH vesicles by E. histolytica is temperature-, microfilament-, and calcium-dependent, and stimulated by phorbol esters.

Notes: . The purpose of this research was to determine whether mice could be protected from lethal challenge with Naegleria fowleri by prior intranasal exposure to pathogenic and nonpathogenic Naegleria. Mortality ranged from 0 to 100% for mice inoculated intranasally (i.n.) with 5 × 103 amebae of 13 human isolates of N. fowleri. Mice were immunized and challenged i.n. using live amebae of strains of low, medium, and high virulence. The greatest protection against lethal challenge was afforded by three immunizing doses of 103 amebae per dose of the strain of medium virulence. Nonpathogenic N. gruberi also was used to immunize mice i.n. against lethal challenge with N. fowleri. Protection was greater following immunization with N. gruberi than it was after immunization with N. fowleri, suggesting that nonpathogenic N. gruberi may be a better immunogen in protecting mice against lethal naeglerial challenge.

Notes: . Stomatogenesis in Opercularia coarctata has been studied in specimens treated with Fernández-Galiano's silver impregnation method. The new buccal structures originate from the germinal row and from the parental haplokinety.

Notes: Bruce-Chwatt, L. J., ed., with Black, R. H., Canfield, C. J., Clyde, D. F., Peters, W. & Wernsdorfer, W. H. 1986. Chemotherapy of Malaria. Peters, W. & Killick-Kendrick, R., eds. 1987. The Leishmaniases in Biology and Medicine, Vol. I: Biology and Epidemiology Euzéby, Jacques. 1987. Protozoologie Médicate Comparée. Les Protozooses des Animaux et Leurs Relations Avec les Protozooses de l'Homme, Vol. II: Myxozoa—Microspora—Ascetospora. Apicomplexa, 1: Coccidioses (Sensu Lato). McDougald, L. R., Joyner, L. P. & Long, P. L., eds. 1986. Research in Avian Coccidiosis. Wichterman, R. 1986. The Biology of Paramecium, 2nd ed. Levine, N. D. & Ivens, V. 1986. The Coccidian Parasites (Protozoa, Apicomplexa) of Artiodactyla.

Notes: . One of the two methanogenic endosymbionts of the giant sapropelic amoeba Pelomyxa palustris was isolated in pure culture. The cells were slender non-motile rods (3 × 0.4 μm), sometimes occurring in chains of 3–4 cells. Ultrathin sections revealed a Gram-positive cell wall and conically pointed ends with mesosome-like structures in the cytoplasm. The isolate had a generation time of 10 and 12 h during growth on H2/CO2 and formate, respectively. The optimum growth temperature was 40°C and the optimum pH was 7.8. The G+C content of its DNA was found to be 37.7% mole percent. The isolate was identified as Methanobacterium formicicum.

Notes: Books reviewed in this article:Larwood, G. & Rosen, B. R., eds. 1979. Biology and Systematics of Colonial Organisms.Hellebust, Johan A. & Craigie, J. S. eds. 1978. Handbook of Phycological Methods. Physiological and Biochemical Methods.

Notes: SYNOPSIS. Surface saccharides in 2 Trichomonas vaginalis strains, the moderately pathogenic, JH34A, and the mild, JH162A, were analyzed with the aid of plant lectins. Concanavalin A (Con A), wheat germ agglutinin (WGA), soybean agglutinin (SBA), castor bean agglutinin (CBA), and lectin from the garden pea (GPA) were employed in agglutination tests and in treatment of ultrathin sections for electron microscopy according to the horseradish peroxidase-3,3′-diaminobenzidine method. With Con A and WGA, small quantitative differences were noted between the 2 strains in the results of agglutination and in the reaction-product deposits observed by electron microscopy. Distribution of the binding sites for the 2 lectins was also somewhat different in the JH34A and JH162A trichomonads. In general, the reactions with the more pathogenic strain were slightly stronger. Although the reactions with SBA and CBA lectins were weaker than those with Con A or WGA, they provided the means for qualitative differentiation between the 2 trichomonad strains. SBA alone agglutinated the JH34A strain and formed demonstrable deposits on the cell surfaces. On the other hand, only CBA reacted with JH162A flagellates. The garden pea lectin failed to bind to the surface of either strain. On the basis of results obtained with the control preparations incubated in the presence of specific inhibitors, it was concluded that both strains had α-methyl-D-mannoside and/or α-methyl-D-mannoside-like as well as N-acetyl-D-glucosamine residues on their surfaces. In addition, JH34A strain had D-lactose-containing residues while JH162A trichomonads had residues with D-galactose. Neither strain appeared to possess residues containing N-acetyl-D-galactosamine.

Notes: SYNOPSIS. Centrifugation for 30–40 seconds at 8,000 g has been used to render monopodial specimens of the large free-living ameba. Chaos carolinensis. These monopodial amebae exhibit obvious torsional movements in the tail. In many cases the posterior ectoplasm assumes the form of a screw with helical ridges forming in place of the more common straight dorsal fins. This finding prompted a re-examination of normal polypodial C. carolinensis, and a majority of these were found also to exhibit torsional movement in the tail and in retracting pseudopodia. These movements suggest that the cytoskeleton of Chaos may have a helical component in its organization.

Notes: Books reviewed in this article:Levandowsky, M. & Hutner, S. H., eds. Biochemistry and Physiology of Protozoa.Raymont, John E. G. with J. D. Burton & K. R. Dyer. 1980. Plankton and Productivity in the Oceans. Vol. I. Phytoplankton.

Notes: Book reviewed in this article:Maramorosch, Karl & Hirumi, Hiroyuki, eds. 1979. Practical Tissue Culture Applications.

Notes: SYNOPSIS. Cysteine and ascorbic acid were previously shown to be required by Entamoeba histolytica trophozoites for attachment to glass, elongation, and ameboid movement as well as for short-term (12–24 h) survival in a balanced salt solution containing bovine serum albumin and a vitamin solution (Maintenance Medium 1). If the only function of cysteine and ascorbate was to decrease the redox potential, other reducing agents should be effective. However, the requirement for cysteine in the presence of ascorbic acid was highly specific. Equally effective were D- and L-cysteine; however, of many other compounds tested, only thioglycolic acid, ascorbic acid, or L-cystine (in decreasing order) were somewhat active. Under N2 atmosphere, cysteine and ascorbic acid were still required, although their concentrations could be halved. The ability to attach in the maintenance medium was irreversibly lost after only 5 min of cysteine-ascorbic acid deprivation; however, there was no decrease in viability when the amebae were transferred to growth medium within 30 min. Cysteine thiol groups in the medium were oxidized rapidly regardless of the concentration of ascorbic acid or the presence of amebae; however, ascorbic acid prolonged attachment of amebae.

Notes: SYNOPSIS. Low concentrations of chlorpromazine (∼0.01 mM) inhibit growth and nucleic acid synthesis in the ciliate Tetrahymena pyriformis. Brief exposure of the cells to, e.g. 0.018 mM chlorpromazine, had very little effect on 14CO2 production or on label incorporation into glycogen from [1-14C]glucetate, [6–14C]glucose, or [1-14C]leucine, but 17-h exposure of stationary phase cultures to this drug caused marked alterations in metabolism, including an almost complete loss of ability to decarboxylate L-[1-14C]leucine and L-[1-14C]tyrosine. It was shown that loss of ability to decarboxylate these amino acids results from loss of ability to transport them.

Notes: SYNOPSIS. A survey of 41 herbivorous mole-rats, Spalax ehrenbergi Nehring, in Urfa, Adiyaman, and Maras provinces of Turkey revealed 7 new species of Eimeria in addition to previously described Eimeriidae. The shape, average dimensions (in μm) of their oocysts, and the numbers of hosts from which the new species were isolated were as follows: Eimeria urfensis sp. n., ellipsoidal (33 × 21), from 8 hosts; Eimeria adiyamanensis sp. n., ovoid to ellipsoidal (33 × 18), from 6 hosts; Eimeria haranica sp. n., elongate ovoid (37 × 20), from 22 rats; Eimeria marasensis sp. n., ellipsoidal (36 × 18), from 2 rats; Eimeria oytuni sp. n., pear-shaped (24 × 17), from 2 hosts; Eimeria celebii sp. n., ellipsoidal (16 × 9), from 1 rat; and Eimeria torosicum sp. n., spherical to subspherical (11 × 10), from 2 animals.

Notes: Defined media are described that support 14-20 h generation times for Acanthamoeba castellanii and A. rhysodes in monolayer cultures. the media differ in minor ways from previously described media, but the growth rates are greatly improved over previously reported values. Maximum growth rates were observed for A, castellanii in a complex medium containing 21 amino acids, but near-maximum rates could be achieved in relatively simple media containing 9 amino acids. Growth occurred with 6 amino acids, as reported by others, but generation times exceeded 30 h. Amitosis was a common problem during early subcultures in defined media, but became infrequent after repeated transfers. Synchronous encystment resulting in 70-80% cyst formation could be induced in the defined media by glucose and acetate starvation. the rate of encystment varied with cell density at the time of starvation and was optimal at initial densities of 400-800 amebae/mm2.

Notes: Tetrahymena pyriformis strain HSM secretes 4 isozymes of hexosaminidase. Purified isozymes B1 and B2 are eluted from the void volume of a concanavalin A-Sepharose column, suggesting that they are not glycosylated. Purified isozymes A1 and A2 bind to the column and are eluted at ∼0.1 M α-methylmannoside, suggesting that these isozymes are glycoproteins. In agreement with earlier deductions based on a differential kinetic assay for the A and B isozymes, the elution pattern of hexosaminidase activity from material secreted by cells grown to early and late stationary phase was consistent with these secretions containing primarily the B and the A isozymes, respectively.

Notes: The surface proteins and glycoproteins on red cells from normal and Babesia bovis-infected calf blood have been compared. Several radiolabeling probes were used to label specifically external membrane molecules which were then separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and identified by autoradiography or fluorography. No differences were observed among the Coomassie Blue-stained membrane proteins of erythrocytes from individual uninfected calves. Comparison of red cells from these animals also indicated no qualitative differences in the surface proteins with accessible tyrosyl residues labeled by lactoperoxidase-catalyzed radioiodnation, although some quantitative variation in the uptake of radioactivity into particular proteins was observed. the major radioiodinated bands on normal bovine erythrocytes had Mr of 165, 130, 90, and 45 kiloDaltons. However, labeling of surface glycoproteins by the periodate/[3H]NaBH4 and galactose oxidase (± neuraminidase)/[3H]NaBH4 methods showed significant differences in the surface proteins of red cells from individual uninfected calves. of 14 animals tested, 5 had major labeled glycoproteins of unique Mr. No changes were observed in radioiodinated surface proteins of total red cell samples from infected calves with 0.5-6% parasitemia. Radioiodination of concentrated infected red cells from the same samples (concentrated by selective hypotonic lysis of uninfected erythrocytes in KC1) resulted in the labeling of 3 new surface proteins, with Mr of 118, 115, and 60 kiloDaltons. the same new 125I-labeled bands were identified on infected cells from 3 avirulent strains of B. bovis used in vaccine production. Furthermore, in concentrated infected cells there was very poor radiolabeling of major bands strongly labeled on uninfected cells (Mr 165, 130, and 90 kiloDaltons), suggesting parasite-induced loss of these proteins. Although there were some differences in 3H-labeled surface glycoproteins of red cells from normal and. B. bovis -infected blood, they were restricted to minor labeled bands and were not seen consistently. the labeled surface glycoproteins of concentrated infected cells were very similar to those of the uninfected red blood cells from infected blood.

Notes: Two kinds of pigment structures, pigment vacuoles and pigmentocysts, cause the orange-red color of Pseudokeronopsis carnea (Cohn, 1866). The pigment vacuoles are undischargeable and two to five layers of them form a characteristic ectoplasmic zone. The pigmentocysts mainly surround the infraciliature and show a unique channel which is probably used for extrusion. Previous data on the fine structure of subpellicular granules and extrusomes of hypotrich ciliates are summarized. Their obviously diverse organization argues for a great value of these structures in species identification. The basic structural features of the infraciliature and the cytoplasmic organelles of P. carnea are similar to those found in other hypotrichs; however, a special kind of linear microtubular array borders the longer sides of the cirral bases and the margins of the adoral membranelles and those of the membranes in the right buccal area. To the left of the endoral membrane, these microtubular arrays result in a highly ordered structure reminiscent of oral ribs. This peculiar arrangement of microtubules in cirri and paramembranelles has also been found in the related form, Thigmokeronopsis jahodai, probably indicating a homogeneity of the fine structure of urostylid hypotrichs. In P. carnea, the basal bodies of the paroral membrane are proximally connected like a polykinetid. Its cilia are unlinked, whereas those of the endoral membrane are fused by microfibrillar material. The terms diplostichomonad and polystichomonad only refer to quantitative aspects and omit the evident, high diversity of microtubular and microfibrillar associates occurring in the membranes in the right buccal area. These terms need to be redefined on the basis of more material that is better described.

Notes: Netzelia tuberculata secretes a test composed of siliceous particles cemented together by organic plaques forming a single-layered spheroidal shell. The siliceous particles are produced within cytoplasmic vacuoles by three mechanisms: 1) synthesis de novo by deposition of the silica on a matrix; 2) deposition of silica on particles remaining in digestive vacuoles, including starch grains and undigested walls of yeast cells; and 3) secretion of silica as a hollow sphere at the periphery of vacuoles enclosed by the silicasecreting membrane. The silicalemma (silica-secreting membrane) originates as fibril-containing vesicles (GFV) secreted by the Golgi body. Fusion of these vesicles with membranes surrounding digestive vacuoles or with membranes surrounding specialized vacuoles containing a silica-binding matrix apparently converts the vacuole into a silica-depositing organelle. Small spherules of silica occur on the vacuolar side of the membrane surrounding the developing test granules, marking the presence of silicalemma activity. These colloidal spherules become aggregated into larger spherules that condense to form the siliceous surface of the developing test particle. Other Golgi vesicles, designated Golgi plaque vesicles (GPV), produce the organic plaques that are deposited among the siliceous particles at the periphery of the cell during new test construction during cell division. The fine structure of the GFV and GPV and their role in test wall deposition are discussed in relation to other silica-biomineralizing protozoa, including radiolaria.

Notes: We have measured binding of fluorescein-conjugated succinyl-concanavalin A (Fl-s-Con A) to bloodstream and procyclic forms of Trypanosoma brucei gambiense and to bloodstream forms of T. b. rhodesiense by flow cytofluorimetry. Bloodstream forms bound an order of magnitude less lectin than procyclic forms. Trypsin-treating cells enhanced binding of Fl-s-Con A to bloodstream forms 3–16-fold depending on the strain and the length of trypsinization but had little effect on Fl-s-Con A binding by procyclics. The trypsinization protocol used did not remove major common glycoproteins detected on lectin blots of either life cycle form but removed 〉95% of the variant specific glycoprotein and fragments derived from this protein of bloodstream forms. Microscopically detectable Fl-s-Con A binding to bloodstream forms was confined to the flagellar pocket. Trypsinized bloodstream forms and procyclics bound Fl-s-Con A in the flagellar pocket, on the flagellum, and on the cell surface. Lectin remained cell associated but appeared to redistribute towards the flagellum and pocket when cells that had bound lectin on ice were subsequently incubated at physiological temperatures. The Fl-s-Con A binding had specificity characteristic of the interaction between the lectin and oligosaccharides. These results are consistent with the hypothesis that the variant specific surface glycoprotein blocks binding of the lectin to surface glycoproteins of bloodstream forms and suggest that concanavalin A-binding glycoproteins are abundant in the flagellar pocket of both life cycle forms.

Notes: The fine structure of the trophozoite and cyst of Entamoeba histolytica from the stool of a patient was compared using the freeze-fracture method. The intramembranous particles (IMP's) were heterogeneously distributed on the plasma membrane of the trophozoite and their density was 1139 ± 105/μm2 on the P face and 27 ± 9/μm2 on the E face. Particle-rich depressions and linear particle arrays, reported by other investigators on cultured trophozoites, were also observed on the P face while on the E face such special particle arrangement was not recognized. Particle-free, small protrusions were frequently observed on the P face of the trophozoite membrane. The existence of these protrusions is a new finding. In the cyst, the IMP's were also distributed heterogeneously on both the P and E faces of the plasma membrane. The density of the IMP's, however, was much lower than in the trophozoite: 6 ± 2/μm2 on the P face and averaging less than 1/μm2 on the E face. In freeze-fracture images, the plasma membrane of the cyst showed a variety of configurations from smooth to uneven or ridged surfaces. These morphological alterations of the plasma membrane may be attributed to the aging of the cyst. The thick wall of the cyst had a filamentous tri- or tetra-lamellar structure. The cytoplasm of the cyst was similar in structure to that of the trophozoite and the diameter of the nuclear pores was equal in both trophozoites and cysts.

Notes: A new starvation procedure permitted the study of early events in a protozoon's growth cycle. Growing cultures of Tetrahymena that differed from non-growing cultures by one variable were produced by adding histidine to cells deprived of that amino acid in an otherwise complete medium. Alterations of the nucleotide pools were examined in +His and in -His cultures in the period preceding RNA synthesis by cells in +His medium. High performance liquid chromatographic analysis provided a balance sheet for the difference in purine compounds in the two cultures. The change in rNTP levels occurred only when the cells were resuspended in a fresh medium and was not a function of cell density. These observations point to the presence of a factor(s) in the old medium that inhibits the energy charge increase in rNTP and in purine accumulation.

Notes: We have recently shown that the ribosomal RNA genes of the amoebo-flagellate Naegleria gruberi Schardinger, 1899, strain NEG-M are carried exclusively on a 14 kilobasepair plasmid. To explore the distribution of this unique gene arrangement, we have examined another strain of N. gruberi and four other species from the order Schizopyrenida. All have this unusual gene arrangement although the size of the plasmid varies widely. Species groups based on morphological criteria do not agree with those resulting from comparison of plasmid restriction enzyme patterns.

Notes: . Sphaerospores were found among three species of fish examined from waters known to be enzootic for proliferative kidney disease (PKD) of salmonids. They were detected in the renal tubules of both hatchery-reared rainbow trout (Salmo gairdneri) exposed to the infectious stage of PKD and in chubs (Gila bicolor) in the headwaters of a hatchery where PKD is enzootic. Sticklebacks (Gasterosteus aculeatus) collected near net pens where Pacific salmon had experienced a PKD epizootic were also found to harbor sphaerospores in the lumen of the kidney tubules. The latter two host species contained developmental stages of a myxosporidan in the blood and in the lumen of the kidney tubules which are similar to those of PKX, the causative agent of PKD in salmonid fish. The sphaerospores observed in the rainbow trout are the first to be observed in this species. The similarity to previously observed developmental stages, rarity, and presence of these sphaerospores in salmonid fish from a hatchery where PKD is enzootic suggest that they are the most mature stage of the PKX myxosporidan yet observed.

Notes: . High-resolution polyacrylamide gradient gel electrophoresis (PGGE) was used to separate isoenzymes of 12 Naegleria strains: one N. australiensis, two N. lovaniensis, one N. jadini, two N. gruberi isolated from environmental samples, and six N. fowleri strains isolated from patients with primary amoebic meningoencephalitis. Of the eight enzymes studied, seven showed zymograms with interspecific variation that identified all the species tested. Although the six N. fowleri strains were biochemically the most homogeneous, they showed intraspecific isoenzyme variation that allowed them to be grouped into four zymodemes. The PGGE technique, which separates isoenzymes by their molecular shape, is both sensitive and economical. It offers an addition or an attractive alternative to isoelectric focusing which has commonly been used to aid species identification of Naegleria by separating isoenzymes by their isoelectric point.

Notes: . In an attempt to solve the ambiguity in the taxonomy of the Euplotes crassus, minuta, and vannus group, 19 strains were tested for mating interactions, electrophoresed for isozymic variations, and analyzed by multivariate morphometrics of the conventional diagnostic traits. The overall results supported the validity of the three named species. Inter-specific mating occurred only in a few crassus x vannus strain combinations and was usually inviable. Isozymic variations, in particular of amylases, malic enzyme, and malic dehydrogenase, were very restricted within conspecific strains and were great between non-conspecifics. The species ascertainment of the strains was possible on the basis of clustering and principal component analyses of morphological measures.

Notes: . Among the known life cycle stages of Trypanosoma cruzi only the amastigote form bound lactoferrin (LF), a glycoprotein produced by neutrophils. This capacity was readily demonstrable by indirect immunofluorescence in amastigotes derived from mice, a mammalian cell culture, or grown in an axenic medium. No LF binding was detectable on trypomastigotes from blood or mammalian cells, insect-derived metacyclics or epimastigotes, or on epimastigotes grown in Warren's medium. Serum levels of LF were increased in mice acutely infected with T. cruzi, and amastigotes from the spleens of these animals were found to have the glycoprotein on their surface. The amastigote LF receptor may have biological significance in parasite-host interaction since mononuclear phagocytes also express a LF receptor, and treatment of these cells with LF has been shown to increase their capacities to take up and kill T. cruzi amastigotes in vitro. The LF receptor is the first marker for T. cruzi amastigotes for which a naturally occurring ligand has been described.

Notes: . The following species are described from Indonesian birds: Isospora paddae n. sp. with oocysts 41.5–45.5 × 40.3–41.5 (44 ± 1.15 × 41.2 ± 0.38) and sporocysts 22.8–24.5 × 14.7–17 (24 ± 0.55 × 16.2 ± 0.81) from the Java sparrow, Padda oryzivora, and Isospora indonesianensis n. sp. with oocysts 39.3–43.6 × 37–40.8 (41.8 ± 1.3 × 39.6 ± 1.25) and sporocysts 25.6–28.4 × 15.2–18.5 (27.1 ± 1.05 × 16.8 ± 1.22) from the chestnut Munia, Lonchura malacca (L.). The host birds belong to the order Passerorida.

Notes: . The composition of the fauna of rumen ciliates in zebu cattle in Kenya was surveyed and 13 genera containing 51 species with 19 formae were identified. Four new species were recognized, then described as Diplodinium africanum n. sp., Diplodinium nanum n. sp., Eudiplodinium kenyensis n. sp., and Ostracodinium iwawoi n. sp. In addition, Buetschlia triciliata and Ostracodinium stokyi were found for the second and third times and described as Hsiungia triciliata n. comb. and Enoploplastron stokyi n. stat., with two formae, respectively. The species composition was similar to that of Indian zebu but several species were considered to originate from African native ruminants. In the percentage composition of genera, the numbers of Entodinium, which normally predominate in rumens in other areas, were very low.

Notes: . Giardia trophozoites and cysts, isolated from mammalian and avian hosts, were examined by transmission electron microscopy (TEM), scanning electron microscopy (SEM), and by fluorescent light microscopy for the presence of microbial symbionts. Mycoplasma-like organisms were observed on the surfaces of trophozoites isolated from the prairie vole, laboratory rat, and beaver. Intracellular bacteria were observed by TEM in the trophozoites and cysts of G. microti and by fluorescence microscopy in trophozoites and cysts of Giardia spp. isolated from beaver, muskrat, great-blue heron, and the green heron. Trophozoites of G. muris from rat small intestine contained viral-like particles measuring 60 nm in diameter. These observations suggest that biological associations between Giardia spp. and diverse microbes may be more common than formerly appreciated. It also raises the possibility of transmission of these apparent symbionts, via the Giardia cyst, to other mammalian hosts including man.

Notes: . Plasmodium falciparum-infecled erythrocytes were metabolically labeled with tritiated glucosamine. Lipid extracts were analyzed by high-performance thin-layer chromatography to compare labeled molecules of eleven isolates from patients, six cytoadherent in vitro strains, and two knobbed and two knobless strains from Aotus monkeys. Up to nineteen labeled bands were identified. Glycolipid GL1, previously identified in Malayan Camp, was present in all isolates and strains. Other molecules, between CG and GM1 and between GM1 and GD1a, varied in mobility or presence. There was no apparent association between labeled molecules and the presence of knobs or the property of cytoadherence.

Notes: Five ciliate species collected from the Woods Hole area were examined by protargol silver impregnation and scanning electron microscopy. These ciliates have been shown to sequester and use chloroplasts obtained from flagellate prey. One new species, Strombidium chlorophilum, is described. Four other species, Strombidium capitatum (Leegaard, 1915) Kahl, 1932, Strombidium conicum (Lohmann, 1908) Wulff, 1919, Strombidium acutum (Leegaard, 1915) Kahl, 1932, and Laboea strobila Lohmann, 1908, are redescribed. Characters used in describing the Strombidiidae include cell size and shape, anterior and ventral polykinetids, macronuclear shape and size, the kinetid “girdle,” the ventral kinety, the trichites, and the paroral kinety. The rationale for using these characters as taxonomic criteria is discussed.

Notes: A new species of coccidium, Tyzzeria chalcides, is described from the ocellated skink, Chalcides ocellatus, from Egypt. Meronts occur in the epithelial cells of the bile ducts and gamonts within the epithelial cells of the gall bladder. Fully sporulated oocysts are cylindrical (L/W ratio 1.88) without a micropyle, oocyst residuum, or polar granule and contain eight spindle-shaped sporozoites and no sporocysts. Sporulation is completed within the gall bladder lumen. Comparison with other species of the genus found in reptiles indicates that it is a new species.

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.