ALBERT

Notes: A method for axenic cultivation of epimastigote and metacyclic forms of Trypanosoma (Duttonella) vivax at 27°C in vitro is described. Iscove's medium was supplemented with specific concentrations of foetal bovine serum, L-proline, L-glutamine, hypoxanthine, adenosine, pyruvate, and 2-mercaptoethanol. Bloodstream form parasites rapidly transformed into epimastigote forms that grew as surface-adherent colonies in plastic culture flasks. Transformation of epimastigotes to metacyclic forms was first observed 9–12 days after initiation of cultures. Percentages of metacyclics varied: East African T. vivax ranged up to 40% and West African T. vivax ranged up to 24%. Subcultures were made at two-week intervals and maintained for several months. Transformation of bloodstream forms to epimastigotes depended on initial attachment to the bottom of culture flasks and the presence of L-proline. The number and maturity of metacyclic forms was influenced by the concentrations of foetal bovine serum, L-proline, L-glutamine, and 2-mercaptoethanol. Trypanosomes from cultures were cryopreserved, revived, and used to re-establish fresh axenic cultures. These results represent a significant advance in cultivation of T. vivax insect forms that should enable studies to be accomplished on metabolism, differentiation, and pharmacology of this parasitic protozoan, free from the influence of extraneous cells.

Notes: We previously reported that phospholipase increases host cell penetration by Toxoplasma gondii. Here we show that calcium-dependent phospholipase A (PLA) activity is found in the supernatant of sonically disrupted T. gondii. When fractions of disrupted T. gondii were incubated with host cells, the release of fatty acids and lysolipids was detected. Fractions of sonically disrupted T. gondii with PLA activity increased T. gondii host cell penetration in a bioassay. In addition, a protein of approximately 20 kDa was detected by immunoblot of T. gondii antigens with horse antiserum to snake venom, the major antibody of which recognizes PLA2. Incubation of T. gondii with exogenous PLA2 resulted in increased solubility of a rhoptry protein. This protein, which we previously characterized as involved with enhanced parasite invasion of host cells and which is recognized by monoclonal antibody Tg49, was detected in increased amounts in supernatant fractions of extracellular parasites treated with PLA2. Whereas without PLA2 treatment, it is only slightly soluble under physiological conditions. This raises the possibility that PLA may be implicated in the release of rhoptry proteins.

Notes: Riboflavin deficiency inhibits the growth of malaria parasites both in vitro and in vivo in infected animals and humans. Although the precise mechanisms underlying this inhibition are unknown, they may involve enhanced requirements for riboflavin by parasites. To investigate this possibility, the rate of uptake of [14C]riboflavin and the biosynthesis of FMN and FAD from riboflavin were studied in infected (5–8% parasitemia) and uninfected human erythrocytes. All cells were incubated for 0–3 h at 37° C in phosphate buffered saline containing MgCl2, glucose, and [14C]riboflavin (2.5–7.5 μM). At hourly intervals, samples were removed, centrifuged, washed twice with cold buffer, and lysed before counting the radioactivity. The rate of in vitro biosynthesis of FMN and FAD from riboflavin in erythrocytes was measured by ion exchange chromatography and reverse isotope dilution techniques. Results showed that the rate of riboflavin uptake and the biosynthesis of FMN and FAD were enhanced in erythrocytes with parasitemia as compared with results in unparasitized erythrocytes. Riboflavin uptake in erythrocytes was proportional to the extent of parasitemia and especially to percent of schizonts present in erythrocytes. These studies indicate that the requirement for riboflavin may be greater in the parasite than in the host erythrocyte. This increased riboflavin requirement may be due to rapid multiplication, higher metabolic rate, and extreme vulnerability to oxidative stress of malaria parasites compared with that of host erythrocytes. The differential requirement of riboflavin by the host and the malaria parasite may hold important potential for developing new strategies for malaria chemotherapy.

Notes: The complete small subunit rRNA gene sequences of the heterotrich Blepharisma americanum and the colpodid Colpoda inflata were determined to be 1719 and 1786 nucleotides respectively. the phylogeny produced by comparisons with other ciliates indicated that C. inflata is allied more closely with the nassophoreans and oligohymenophoreans than the spirotrichs. This is consistent with the placement of the colpodids in the Class Copodea. Blepharisma americanum was not grouped with the hypotrichs but instead was placed as the earliest branching ciliate. the distinct separation of B. americanum supports the elevation to class status given the heterotrichs based on morphological characters.

Notes: Endosymbiotic methanogenic bacteria of three species of anaerobic ciliates (Plagiopyla frontata, Metopus conforms, and M. palaeformis) were inactivated with the specific methanogen inhibitor 2-bromoethanesulfonic acid. the absence of endosymbiont methanogens reduced growth rate and growth yield by about 30% in P. frontata and M. contortus, while no significant change in fitness was observed in M. palaeformis. In Plagiopyla the growth rate constant is not affected by an artificially increased pH2 neither in normal nor in methanogen-free ciliates. the energetic advantage conferred by endosymbiont methanogens in Plagiopyla and in Metopus contortus probably is due to excretion of organic material from the bacteria at the expense of bacterial reproduction. It is unlikely that the maintenance of a low pH2 within the cells due to H2-consumption by the bacteria is important to the ciliates.

Notes: Feces from a juvenile specimen of the anteater Tamandua tetradactyla from Ponta de Pedras, Marajó, Pará, northern Brazil, contained three different coccidial oocysts: Eimeria tamanduae Lainson, 1968; E. corticulata Lainson & Shaw, 1990; and a third species previously unrecorded and described here as Eimeria marajoensis n. sp. Oocysts of the latter parasite are spherical to subspherical, 13.9 ± 1.5 times 13.4 ± 1.4 (11.1-16.5 times 11.1-16.5) μm, shape index (length/width) 1.0 (1.0-1.2). the oocyst wall is a single, Colorless layer about 0.6-1.0 μm thick with no striations or micropyle. There is no oocyst residuum, but a single, round, oval or irregularly shaped polar granule of about 0.75-2.5 μm is consistently present. the sporocysts are broadly ellipsoidal, 7.1 ± 0.7 ± 5.3 ± 0.6 (6.0-8.8 times 4.0-5.7) μm, shape index 1.3 (1.2-1.5), with a delicate wall bearing a minute stieda body. No sub-stieda body was visible. the sporocyst residuum consists of some 10-20 rounded granules, lying between the two slightly curved sporozoites which measure approximately 6.5 times 2.0 μm. Sporocyst refractile bodies were not discernablc.

Notes: Exoerythrocytic forms of Plasmodium gallinaceum were cultured in vitro using salivary gland sporozoites extracted from experimentally infected Aedes fluviatilis mosquitoes. the host cells were macrophage precursors from chicken bone marrow. At various times after introduction of Sporozoites, the cultures were stained by Giemsa or by immunofluorescence assay (IFA) using anti-sporozoite-specific monoclonal antibodies (MAb). the time to complete parasite development in vitro was 50-70 h. By 70 h, ruptured segmenters and free merozoites were visible within the cells. Inoculation of normal chickens with infected cultures induced parasitemia after a pre-patent period of 10-11 days. In vitro young exoerythrocytic forms, late schizonts that include the matured segmenters, and free merozoites shared common antigens with the sporozoites as revealed by IFA using anti-sporozoite-specific MAbs. Our data indicate that macrophages support development of P. gallinaceum sporozoites and that the circumsporozoite proteins are present until Ac end of the primary exoerythrocytic schizogony.

Notes: This past-presidential address considers the relationship between small-scale assembly and large-scale patterning in ciliates. Certain model examples of assembly-based patterning are reviewed, followed by the introduction of Williams and Honts' concept of “meta-assembly” as applied to the development of the oral apparatus. The major part of this review then explores two topics: the nature and origin of large-scale circumferential order, and the manner by which the large-scale order of cell directions influences the organization of membranelles of the oral apparatus. In this review, I summarize an existing formal description of large-scale positional order, allude to a more precise abstract theoretical model, and end with a brief discussion of the problem of searching for molecular mechanisms.

Notes: Ellobiophrya conviva clasps tentacles of the bryozoan Bugula neritina with a ring-like structure formed from aboral extensions of its body that taper into two slender arms. The tips of the arms overlap and join to form a unique organelle, the bouton. Each arm contains a massive myoneme that splays out at the bouton. The bouton consists of the cupped tips of the arms and a cavity, which is filled with dense homogeneous material. Long digitations containing longitudinal microtubules at their periphery project from the inner surface of the tip of each arm into the cavity. Deep folds of pellicle with pores opening into their depths line the wall of the cavity. Conventional kinetosomes are not visible in the bouton, but circular or elliptical arrays of microtubules are found at the bases of digitations. The nonfunctional scopula of the adult is in a depression enclosed by pellicular folds. The bouton is distant from the scopula, but its fine structure somewhat resembles it, supporting Chatton and Lwoff's hypothesis that the cinctal arms carry parts of the scopula at their tips. The fine structure of the cinctum supports their suggestion that the cinctal arms are homologous to the spasmonemes of vorticellid peritrichs.

Notes: Twenty monoclonal antibodies were produced against trophozoites of Entamoeba histolytica strains HK-9 and HM-1: IMSS. When reactivity to various enteric protozoa was examined by an indirect fluorescence antibody test, 15 of the monoclonal antibodies were strongly reactive with E. histolytica trophozoites. Species-specific antigens recognized by these monoclonal antibodies were located on the plasma membrane, nucleus, cytoplasm, and cytoskeletal structures of the trophozoites. Two of the remaining five monoclonals reacted strongly with trophozoites of the E. histolytica-like Laredo strain. The determinant antigen was located in the cytoplasm. The three remaining monoclonal antibodies were found to recognize cross-reactive antigens between E. histolytica and E. histolytica-like Laredo, E. hartmanni, E. coli, Dientamoeba fragilis, Giardia lamblia, and Trichomonas hominis. These three antibodies were also reactive with T. vaginalis

Notes: . Macrostomal cell formation is blocked by the antibiotic cerulenin at levels of 15 μg/ml or higher. Inhibition can be reversed up to 4 h following cerulenin addition by washing and resuspending cells in new, noncerulenin-treated transforming principle. In these latter cases, additional time equal to the time spent in the inhibitor, is needed for cells to reach control values of transformation. Neither the addition of saturated or unsaturated fatty acids, cholesterol added alone or in combination with stearic acid, nor a mixture of lipids extracted from Tetrahymena vorax reversed the cerulenin effect. Radioisotope incorporation data showed while protein synthesis was reduced by the end of 1 h and tetrahymanol synthesis by the end of 2 h, little or no effect of this inhibitor occurred on RNA or fatty acid synthesis during these times. One interpretation of these results is that cerulenin, by preventing first protein synthesis and later tetrahymanol synthesis, interferes with synthesis and formation of membranes required for the microstome to macrostome transition.

Notes: Microfilum lutjani n. g., n. sp. (Microsporida) was found on the gill filaments of Lutjanus fulgens (Teleost) inhabiting the coasts of Senegal. This microsporidium forms xenomas distinguished by the microvilli covering the plasma membrane. At all stages of development individuals have isolated nuclei and are in direct contact with the host cytoplasm. Merogony is binary and sporogony is tetrasporoblastic. the spore (4.75 times 2.60 μm)) is characterized by a manubrium inserted on a laterally offset anchoring disc and extending into a very short, noncoiled polar filament (no longer than 500 nm) in the form of a hook. This type of polar filament has not been described previously in the Microsporida.

Notes: Geosim was identified as the cause of a distinct earthy/grassy odour detected in cultures of a free-living amoeba, Vannella species. Volatile components of cell lysates were isolated and concentrated by the Closed Loop Stripping method. Capillary, gas chromatography/mass spectrometry was used to identify odorous compounds. Bacterial symbionts observed in the cytoplasm of the amoebae may be responsible for production of the geosmin. This appears to be the first report of odorous compounds associated with a free-living protozoan and suggests that in some circumstances, Vannella sp. may contribute to taste and odour problems in drinking water.

Notes: The literature on discriminant feeding by planktonic protozoans using geometric and nongeometric criteria is reviewed with emphasis on recent studies that indicate phagotrophic protists can use information other than particle size or shape to sort among potential prey. Sufficient data are available for ciliates, aplastidic microflagellates, and phagotrophic dinoflagellates. Numerous representative taxa of all three groups have chemosensory capabilities, either to specific chemicals or to prey exudates, that modify their motility patterns resulting in aggregation or dispersal. Representatives of all three groups also have specific prey preferences. These considerations imply, but do not prove, selectivity in feeding through use of chemical cues. Although prey geometry is clearly a first-order determinant of ingestion through passive mechanical selection, recent studies illustrate that planktonic ciliates and flagellates can use other criteria to discriminate among prey. the evidence clearly implicates use of chemical cues, most likely perceived through contact chemoreception. Filter feeders as well as raptors have such abilities indicating that feeding mechanisms per se do not imply limitations on feeding behavior. Evidence of considerable flexibility and complexity in chemoperceptive feeding suggests that we have only glimpsed the more detailed features of feeding behavior in aquatic protozoans.

Notes: The study of microbial food webs is dominated by field measurements of microbial standing stocks and rate processes and to a lesser extent by laboratory studies. These approaches reflect the concerns of microbial ecologists to assess accurately the capabilities of microorganisms and to compare microbial processes to other ecosystem parameters. These approaches have led to enormous advances in understanding microbial food webs. Reconciling our expanding knowledge with general questions about the significance and representation of microbial food webs in ecosystem studies requires additional approaches including comparative studies and field experiments. Comparative studies, analyses of microbial stocks or rates across a wide range of ecosystems, lead to quantitative models of microbial processes. These models facilitate testing of hypotheses at a very general level, allow the comparison of different stocks or rate processes across a gradient of systems, and detect unusual situations or outlier systems. Field experimental manipulations offer the advantages of working with intact natural communities, of direct evaluation of results with statistical methods, and of testing important qualitative hypotheses. Both comparative and field manipulation studies have led to important advances in the study of microbial food webs and should be expanded.

Notes: Repeated exposure of trypanosomes in vitro or in vivo to low concentrations of the methylating agent 1,2-bis(methylsulfonyl)-1-methylhydrazine induces a series of moderately synchronous morphological and biochemical changes. Cell division halts and the long-slender bloodstream forms transform to short-stumpy forms via larger intermediate-stage cells which contain approximately double the normal G2 content of DNA. In common with naturally occurring short-stumpy trypanosomes, drug-induced short-stumpy forms do not infect rodents and when transferred to Cunningham's medium, transform to and replicate as procylics. Furthermore, these short-stumpy forms exhibit α-ketoglutarate supported motility and oxygen consumption, acquire the ability to reduce nitroblue tetrazolium (NADH diaphorase positivity) and appear to be in the G1 or G0 stage of the cell cycle based upon DNA content.

Notes: Tomonts and their theront offspring of the hymenostomatid fish parasite Ichthyophthirius multifiliis were exposed to calcium levels from 0 to 0.8 mM Ca2+. The survival and reproductive rates of tomonts in the absence of extracellular calcium were not significantly different from rates of tomonts provided calcium. Theronts that developed in the absence of calcium, however, were not infective for Ictalurus punctatus even when the extracellular magnesium concentration was doubled. Theronts that developed in 0.10 mM Ca2+ were infective (0.77 trophonts/mm2 of pectoral fin) to essentially the same extent as theronts provided 0.33 mM Ca2+. Infectivity of those provided 0.8 mM Ca2+ was 1.79 trophonts/mm2 of fin, similar to that of theront controls. Theronts deprived of extracellular calcium as they developed contained significantly fewer secretory mucocysts than did theronts provided 0.1 to 0.8 mM Ca2+ although no significant differences among groups occurred with respect to abundance of crystalline or differentiating mucocysts. Theronts deprived of extracellular calcium also had swollen or enlarged mitochondria and abnormal crystalline mucocysts.

Notes: Four types of anomalous conjugation were documented in Stylonychia mytilus. Type I pairs were formed between mates of different sizes. These pairs exhibited an abnormal site of fusion in at least one of the mates, and the mates might face each other ventrally throughout conjugation instead of the normal side-by-side position. Type I pairs underwent sexual nuclear development and proceeded with the first cortical reorganization as in normal conjugants. Type II involved pairing at the anterior ends of mates with ventral surfaces facing the same direction. These pairs also underwent sexual nuclear development. Hence, aberrant orientation of the mates, and also ectopic sites of cytoplasmic fusion, if extensive, would permit sexual development. Type III pairs were united ventral-to-ventral with their anterior-left sides at the adoral zone of membranelles, and remained as such throughout conjugation. In these pairs, nuclear and cortical events were typical of the asexual development of physiological reorganization. In Type IV pairs, one mate of the pair possessed a fission furrow and developed two sets of ciliature typical of binary fission, while the other mate might undergo physiological reorganization or binary fission. Type III and Type IV pairs thus reveal the asexual state of early conjugants, which can pursue either one of the two modes of asexual cortical reorganization; these cases reinforce the notion of overlap of asexual and sexual cycles during conjugation of hypotrichs. Spontaneous autogamy was documented for the first time for this genus. The autogamonts proceeded with nuclear development and with the first cortical reorganization. Some probably underwent second and third reorganizations, as in conjugants, but accompanied by abnormalities, particularly in the stages beyond fertilization. Post-autogamous clones were nonviable except for one dubious case.

Notes: Leishmania major promastigotes were washed and resuspended in an iso-osmotic buffer. The rate of oxidation of 14C-labeled substrates was then measured as a function of osmolality. An acute decrease in osmolality (achieved by adding H2O to the cell suspension) caused an increase in the rates of 14CO2 production from [6-14C]glucose and, to a lesser extent, from [1, (3)-14C]glycerol. An acute increase in osmolality (achieved by adding NaCl, KCl, or mannitol) strongly inhibited the rates of 14CO2 production from [1-: 14C]alanine, [1-14C]glutamate, and [1, (3)-14C]glycerol. The rates of 14CO2 formation from [1-14C]laurate, [1-14C]acetate, and [2-14C]glucose (all of which form [1-14C]acetyl CoA prior to oxidation) were also inhibited, but less strongly, by increasing osmolality. These data suggest that with increasing osmolality there is an inhibition of mitochondrial oxidative capacity, which could facilitate the increase in alanine pool size that occurs in response to hyper-osmotic stress. Similarly, an increase in oxidative capacity would help prevent a rebuild up of the alanine pool after its rapid loss to the medium in response to hypo-osmotic stress.

Notes: The developmental stages of a recently described microsporidian from the nucleus of hematopoietic cells of salmonid fish were found to be unique among the Microsporida. All observed stages, including meronts, sporonts, and spores were in direct contact with the host cell nucleus (principally hematopoietic cells) of chinook salmon (Oncorhynchus tshawytscha). There is no par-asitophorous vacuole and sporogony does not involve formation of a pansporoblastic membrane as with other members of the suborder Apansporoblastina. The extrusion apparatus differentiates prior to division of sporogonial plasmodia. The spores are ovoid (1 times 2 μm) and uninucleate, and possess a coiled polar tube with 8-12 turns. Developmental stages of the salmonid microsporidian are similar to those described for Enterocytozoon bieneusi as found in the intestinal mucosa of human AIDS patients. However, the intranuclear development, different cell types, and host infected clearly separate the salmonid and human parasites. Accordingly, the intranuclear parasite of salmonids is given the name Enterocytozoon salmonis n. sp. within the suborder Apansporoblastina.

Notes: . Mirror-image symmetry doublets of the ciliate Stylonychia pustulata were obtained from the progenies of dividing cells in which cell division was inhibited by heat-shocks. In two components consisting of the doublet, the left (cell's) component possessed ciliary organelles arranged in almost the same pattern as in normal singlets, while the right one had surface organelles located in a mirror-image symmetry of those of the left component. In cell division of the doublet, two sets of ciliary primordia that were arranged in a mirror-image symmetry developed synchronously in both components. In about 80% of oral primordia (OP) of the right components, the arrangement of the membranellar bands became abnormal. In some cases, OP of the right component were occasionally separated into two longitudinal halves, each consisting of normal membranelles and inverted membranelles. A set of primordia of the paroral membranelles and fronto-ventro-transverse cirri was rarely derived from the basal bodies of the right half with a band of normal membranelles. As a result, a third component with the ciliary organelles normally arranged emerged on the right side of the original right component. The differentiation of membranelles and segmentation of the primordial streaks into cim proceeded from anterior to posterior. A cytoplasmic bulge with multiple right marginal cirral rows was frequently formed at the right margin of the doublet. The behavior in the separation of third and fourth streaks from a primordial streak of dorsal cirri was not mirror-image symmetrical in each component.

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

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

Notes: 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: . Major fatty acid components of Acanthamoeba castellanii lipids extracted after growth at 30°C include myristate, palmitate, stearate and the polyunsaturates linoleate, eicosadienoate, eicosatrienoate and arachidonate, with oleate as the sole major monounsaturated fatty acid. By comparison, growth at 15°C gave increased linoleate, eicosatrienoate and arachidonate, but decreased oleate and palmitate. When the growth temperature was shifted downwards from 30°C to 15°C, increased lipid unsaturation occurred over a period of 24 h; thus decreases of oleate and eicosadienoate were accompanied by increases in linoleate, eicosatrienoate, arachidonate and eicosapentaenoate. An upwards shift from 15°C to 30°C gave negligible alterations in fatty acid composition over a similar period. At 15°C organisms rapidly use [1-14C] acetate for de novo fatty acid synthesis; stearate is converted via oleate to further desaturation and chain elongation products. Similar short term experiments at 30°C indicate only de novo synthesis and Δ9-desaturation; synthesis of polyunsaturates was a much slower process. Rapid incorporation of [1-14C] oleate at 30°C was not accompanied by metabolic conversion over two hours, whereas at 15°C n-6 desaturation to linoleate was observed. Temperature shift of organisms from 15°C to 30°C in the presence of [1-14C] acetate revealed that over half of the fatty acids in newly-synthesised lipids were saturated, but the proportions of unsaturated fatty acids increased with time until the total polyenoate components reached 17% after 22 h. A shift of temperature in the reverse direction gave a corresponding figure of 60% for polyunsaturated fatty acids. These results emphasize the importance of n-6 desaturation in the low temperature adaptation of Acanthamoeba castellanii.

Notes: . A survey for Babesia microti in rodents was conducted at six sites within Grand Teton National Park, Wyoming. Blood and spleen smears, hematocrits, and reticulocyte counts were made on all of the animals to evaluate parameters for the diagnosis of babesiosis. Ticks were removed for identification. Of 257 Microtus montanus, 103 were infected with B. microti. In addition, five of 12 Microtus pennsylvanicus and one of three Arvicola richardsoni were parasitized by B. microti. Peromyscus maniculatus (n = 40) were not infected. Concurrent infections by Hepatozoon sp., Trypanosoma sp., and the bacterium, Grahamella sp., were noted in blood smears from a number of M. montanus. Splenomegaly and reticulocytosis were significant parameters associated with babesiosis while decreased hematocrit was not. Ticks removed from the voles were identified as Ixodes eastoni and were the probable vectors of the B. microti.

Notes: . Analysis of total DNA isolated from the Chrysophyte alga Ochromonas danica revealed, in addition to nuclear DNA, two genomes present as numerous copies per cell. The larger genome (˜120 kilobase pairs or kbp) is the plastid DNA, which is identified by its hybridization to plasmids containing sequences for the photosynthesis genes rbcL, psbA, and psbC. The smaller genome (40 kbp) is the mitochondrial genome as identified by its hybridization with plasmids containing gene sequences of plant cytochrome oxidase subunits I and II. Both the 120- and 40-kbp genomes contain genes for the small and large subunits of rDNA. The mitochondrial genome is linear with terminal inverted repeats of about 1.6 kbp. Two other morphologically similar species were examined, Ochromonas minuta and Poteriochromonas malhamensis. All three species have linear mitochondrial DNA of 40 kbp. Comparisons of endonuclease restriction-fragment patterns of the mitochondrial and chloroplast DNAs as well as those of their nuclear rDNA repeats failed to reveal any fragment shared by any two of the species. Likewise, no common fragment size was detected by hybridization with plasmids containing heterologous DNA or with total mitochondrial DNA of O. danica; these observations support the taxonomic assignment of these three organisms to different species. The Ochromonas mitochondrial genomes are the first identified in the chlorophyll a/c group of algae. Combining these results with electron microscopic observations of putative mitochondrial genomes reported for other chromophytes and published molecular studies of other algal groups suggests that all classes of eukaryote algae may have mitochondrial genomes 〈 100 kbp in size, more like other protistans than land plants.

Notes: Earlier studies showed that Leishmania major promastigotes are sensitive to osmotic conditions. A reduction in osmolality caused the cells to shorten and to rapidly release most of their large internal pool of alanine. In this study some effects of hyper-osmotic stress were examined. an increase in osmolality of the culture medium from 308 to 625 mOsm/kg caused only a small decrease in growth rate. When cells grown in the usual culture medium (308 mOsm/kg) were washed, resuspended in iso-osmotic buffer, and subjected to acute hyper-osmotic stress by addition of mannitol, the alanine content increased even in the absence of exogenous substrate. Promastigotes, depleted of alanine by a 5-min exposure to hypo-osmotic conditions, also synthesized alanine when resuspended in iso-osmotic buffer. Washed cells resuspended in iso-osmotic buffer consume their internal pool of alanine under aerobic conditions, Rates of consumption decreased on addition of mannitol, becoming zero at about 440 mOsm/kg. At higher osmolalities, alanine synthesis occurred. to estimate whether proteolysis could account for alanine synthesis in the absence of exogenous substrate, cells that had been grown with [1-14C]leucine were washed and resuspended under hypo-, iso-, and hyper-osmotic conditions and the amounts of 14CO2 and 14C-labelled peptides released in 1 h were measured. Little proteolysis occurred under these conditions, but the possibility that proteolysis was the source of the alanine increase, observed in response to hyper-osmotic stress, cannot be ruled out.

Notes: The evidence for a qualitatively and quantitatively important trophic link between planktonic Protozoa and higher order metazoan consumers is reviewed. the available data are obtained primarily, but not exclusively, from laboratory studies of calanoid copepod consumers and tintinnid ciliate prey from marine estuarine and nearshore environments. the data indicates that the protozoan-metazoan link is of similar magnitude and importance in the pelagic ecosystems of freshwaters. It is proposed that planktonic Protozoa constitute a high quality, nitrogen-rich food in the diets of their metazoan consumers. Implications of die trophic link to the consumers, prey, and ecosystem are discussed.

Notes: A protein with two subtypes of 205 and 180 kDa was localized on the nuclear envelope of amoebae as detected by indirect immunofluorescence staining and immuno-electron microscopy using a monoclonal antibody as a probe. Electron microscopic observation showed that the protein was located on the honeycomb lamina of the nuclear envelope. During mitosis, the protein dispersed throughout the cytoplasm but reappeared on the nuclear envelope after the reformation of the envelopes of daughter nuclei. the findings suggested that the protein is a component of the nuclear lamina of amoebae.

Notes: This study demonstrates that Pleistophora schubergi Zwölfer, 1927, a microsporidium originally isolated from the midgut epithelium of Nygmia phaeorrhoea Don (Euproctis chrysorrhoea L.) and Porthetria dispar L., and subsequently reported in several other insects including the spruce budworm, Choristoneura fumiferana (the host used in this investigation), does not belong in the genus Pleistophora Gurley, 1893. Pleistophora schubergi lacks the major features that are characteristic of Pleistophora typicalis, the type species of this genus. A comparison of ultrastructural observations reported for the type species of the genus Pleistophora, P. typicalis, and our observations of P. schubergi revealed significant differences. A thick (0.5 μm) amorphous coat, derived from parasite secretions and deposited external to the parasite plasmalemma, surrounds all developmental stages in P. typicalis. Double membranes, derived from host rough endoplasmic reticulum cisternae encircle the parasite plasmalemma of all developmental stages in P. schubergi. The sporophorous vesicle encases the spores in P. typicalis, and originates from the parasite-secreted coat that is present around meronts. In P. schubergi, the host endoplasmic reticulum cisternae form the envelope that surrounds the meronts. Moreover, the sporophorous vesicle envelope in P. typicalis persists around groups of spores, while in P. schubergi this envelope breaks easily to release the spores in the host cytoplasm. By comparing the characteristics of the microsporidium found in the spruce budworm with those of the recently created polysporous genera that sporulate within a vesicle, we found that P. schubergi does belong in the new genus Endoreticulatus Brooks et al. 1988, and consequently rename it Endoreticulatus schubergi (Zwölfer, 1927) n. comb.

Notes: Some protists from both marine and freshwater environments function at more than one trophic level by combining photosynthesis and panicle ingestion. Photosynthetic algae from several taxa (most commonly chrysomonads and dinoflagellates) have been reported to ingest living prey or nonliving particles, presumably obtaining part of their carbon and/or nutrients from phagocytosis. Conversely, some ciliates and sarcodines sequester chloroplasts after ingestion of algal prey. Plastid retention or “chloroplast symbiosis” by protists was first demonstrated 〈 20 years ago in a benthic foraminiferan. Although chloroplasts do not divide within these mixotrophic protists, they continue to function photosynthetically and may contribute to nutrition. Sarcodines and ciliates that harbor endosymbiotic algae could be considered mixotrophic but are not covered in detail here. the role of mixotrophy in the growth of protists and the impact of their grazing on prey populations have received increasing attention. Mixotrophic protists vary in their photosynthetic and ingestion capabilities, and thus, in the relative contribution of photosynthesis and phagotrophy to their nutrition. Abundant in both marine and freshwaters, they are potentially important predators of algae and bacteria in some systems. Mixotrophy may make a stronger link between the microbial and classic planktonic food webs by increasing trophic efficiency.

Notes: The lorica of the tectiform choanoflagellate D. costata contains five categories of costal strips distinguishable from each other on the basis of morphology and patterning. Categories of strips include those forming the anterior transverse costa; the anterior, intermediate, and posterior costal strips, respectively, of the longitudinal costae and those constituting the posterior transverse costa. the distinctive morphology of each class of strips makes it possible to observe their location and orientation within the overall accumulation of strips at the top of the parent cell collar. In Diplotheca costata the orientation and positioning of the different categories of strips in an accumulation anticipates their orientation and imbrication in the mature lorica. Assembly of the lorica from an accumulation of strips involves lateral sliding of costal strips to constitute transverse costae and longitudinal sliding of strips to constitute longitudinal costae. the motive force for lorica assembly is provided by extension of the anterolateral tentacles.

Notes: . The light microscopic and ultrastructural characteristics of a microsporidium provisionally identified as Toxoglugea chironomi (Debaiseux, 1931) Jírovec, 1936, is described. It was isolated from oenocytes and adipose tissue of a midge larva of the genus Dicrotendipes. Merozoites are diplokaryotic. The sporogony produces, by fragmentation, eight monokaryotic spores in a sporophorous vesicle. Mature spores are horse-shoe shaped. The total length is about 5.8 μm, the width 0.8-0.9 μm, the external height of the curve 2.3-3.5 μm, and the external width of the curve 3.5-5.2 μm. The polaroplast has lamellar compartments of two types: narrow and closely packed anteriorly, and wider and more loosely arranged posteriorly. The isofilar polar filament is arranged in 8–10 coils in the posterior fourth of the spore. The external nuclear membrane is sometimes continuous with the endoplasmic reticulum. Lamellar and tubular material of exospore construction are present in the episporontal space from the beginning of sporogony. Teratological and normal spores sometimes occur together in the sporophorous vesicle. The identification of the species is discussed and the ultrastructure is compared to Toxoglugea variabilis, the only further species of the genus with known ultrastructural cytology.

Notes: . Paramecium calkinsi from tidal marshes survive a wide salinity range. Fluid output of contractile vacuoles of these cells decreased as salinity of the medium to which they were acclimated increased, and both pulse rate and vacuole volume were used to regulate output. When cells were first exposed to more dilute medium, contractile vacuoles greatly increased volume so that fluid output increased even though pulse rate decreased. In cells shifted to a more concentrated medium, contractile vacuole output decreased by decreasing pulse rate. The contractile vacuole is surrounded by a set of collecting structures which change form as the salinity changes. Distensible ampullae are found in media of low salinity and collecting canals are found in media of high salinity. When cells are shifted from high salinity to low, the number of ampullae increases and the number of canals decreases. When cells are shifted from low salinity to high, the number of ampullae decreases and the number of canals decreases. Other non-contracting vacuoles also appear in response to a hypoosmotic shock. These include vacuoles within the cell as well as “blisters” on the surface. The number and frequency of blisters increases with the size of the hypoosmotic shock. They detach from cells without resulting in any visible loss of cytoplasm. Non-contractile vacuoles may play a role in sequestering and removing excess water that the contractile vacuoles cannot handle.

Notes: The gregarine Cosmetophilus vonones, n. g., n. sp. (Sporozoasida: Actinocephalidae: Acanthosporinae) from the intestine and intestinal caeca of the harvestman Vonones sayi (Simon) (Opiliones: Cosmetidae) is described. The new internal parasite is the first recorded from a harvestman in the New World and the second from the opilion suborder Laniatores. In addition to the records from the type locality in western Texas, additional collections are reported from the same host in central and eastern Texas and Tennessee.

Notes: The morphology, infraciliature, and life cycle of Endosphaera terebrans, a suctorian endocommensal of peritrichs, have been studied with the aid of silver impregnation.The life cycle of Endosphaera terebrans begins with infection of the host cell by a small larva. The swarmer has a pointed needle-like cellular projection and two rings of cilia. The swarmer penetrates the peritrich, loses the cilia, and then matures into an adult. The infraciliature of the adult form has four rows of barren kinetosomes that lack kinetodesmal fibers. By endogenous budding, a migratory larva is produced that leaves the host cell through the peristomial disc and that can infect other peritrichs.

Notes: The strain N of Trimyema compressum, an anaerobic free-living ciliate, was cultivated axenically in a medium containing a buffered salt solution, yeast extract, trypticase, and glutathione. Dead bacteria were indispensable as food; a culture of the ciliate together with heat-killed Klebsiella pneumoniae has been established for more than one year. In the medium described, the ciliates grow to a higher cell density than in cultures with living bacteria as food. During the process of axenization, a nonmethanogenic bacterial endosymbiont was lost. In the microbodies of T. compressum, hydrogenase could be localized by the technique of indirect immunofluorescence.

Notes: Encephalitozoon hellem is a new human microsporidian isolated from corneal biopsies and conjunctival scrapings of three AIDS patients and cultured in Madin Darby canine kidney (MDCK) cells. Encephalitozoon hellem and Encephalitozoon cuniculi display different protein profiles with sodium dodecyl sulfate-polyacrylamide gel electrophoresis and unique antibody binding patterns with murine antisera against Western blots of each organism. Developmental stages of E. hellem in culture are similar to E. cuniculi. Meronts are 1.3–2.7 μm in diameter, develop within a parasitophorous vacuole adjacent to the vacuolar membrane, divide by binary fission, and contain one or two discrete nuclei. Sporonts measure 2 × 3 μm, separate from the vacuolar membrane, and have a thickened outer membrane. Sporoblasts display a tri-layered wall and possess the earliest recognized polar filaments. Mature spores measure 1 × 1.5 μm and are more electron-dense than other stages. Each spore contains a single nucleus, a polar tubule with four to nine coils, thin electron-dense exospore and thick, electron-lucent endospore. Although E. hellem and E. cuniculi differ biochemically and immunologically, their fine structure and development are indistinguishable.

Notes: The effect of the microtubule inhibitor nocodazole was studied on Paramecium and shown to arrest cell multiplication, depolymerize the internal microtubule network, and block the development of macro- and micronuclear spindles and of the cytospindle (a cortical microtubule array assembled during division). After ultraviolet mutagenesis, three mutants resistant to nocodazole, that is capable of continued growth in the presence of the drug, were isolated and shown to correspond to three nonallelic single-gene nuclear mutations. One (nocr-1) is semidominant while the other two (nocr-2 and nocr-3) are recessive. Cytological and physiological studies of nocodazole's effects on the mutants demonstrate that their resistance is due neither to a lack of drug penetration nor to its degradation since, in each mutant in the presence of the drug, some microtubule networks are normal or subnormal while others remain affected as in wild-type cells. These are the first mutants resistant to microtubule depolymerizing drugs obtained in ciliates that provide a new tool for studying the assembly and dynamics of the diverse microtubule arrays in this type of organism.

Notes: We observed a wide distribution of the carbohydrate epitopes galactosylα(1–3) galactose (galα1–3 gal), α-glucoside, and α-mannoside in mono- and heteroxenic trypanosomatids by using fluorescein-labelled lectins of Euonymus europaeus (EE) and Concanavalin A (Con A) as well as sera from acute chagasic patients who have very high levels of anti-galα(1–3) gal antibodies. The direct fluorescence test for galα1–3 gal with EE was positive at minimum concentrations of 6 μg/ml for heteroxenic trypanosomatids and 0.7 μg/ml for monoxenic ones and for the plant parasite, Phytomonas. On the other hand, heteroxenic trypanosomatids that infect vertebrates bound ten-fold more Con A than monoxenic flagellates and Phytomonas. These data were confirmed in ELISA and Western Blot assays carried out with peroxidase-labelled EE and Con A. Euonymus europaeus recognized several glycoproteins in all trypanosomatids that we tested. Con A, however, recognized a glycoprotein cluster in heteroxenic protozoa, which ranging from 60–120 kDa, seemed to lack monoxenic parasites and Phytomonas. These findings suggest that α-D-mannose and α-D-glucose might play an important role in the interaction between trypanosomatids and vertebrate hosts.

Notes: Nuclear and cortical phenomena during dividing and resting cyst formation of Colpoda inflata are described. Cell division forms a cyst and produces two or four tomites. In each tomite, the right oral field results from the proliferation of the anterior extreme of a single kinety, and the left oral field results from the proliferation of four, five, or six somatic kineties. After macronuclear division, each macronuclear mass undergoes a chromatinic extrusion process. During resting cyst formation, the oral infraciliature of the vegetative cell is resorbed. The somatic kineties dispose in a radial way and some pairs of kinetosomes disappear. As in cell division, there is an extrusion process. From these results we conclude that the resting cysts of Colpoda inflata cannot be included in any group of the previous classifications for hypotrich resting cysts. Thus, we propose a new additional group to Walker and Maugel's classification called PKR (partial-kinetosome-resorbing) cysts.

Notes: The proteinase activity present in homogenates of trophozoites of Giardia lamblia, active on azocasein and urea-denaturated hemoglobin, was separated into two different enzymes by a series of purification procedures. These procedures included gel filtration on Fractogel TSK HW-55 (F), organomercurial agarose affinity chromatography, and ion exchange chromatography on DEAE-cellulose. By chromatography on Sephadex G-100, two purified enzymes exhibited relative molecular weights of Mr= 95,000 and 35,000 ± 10%, respectively. On the basis of inhibition by thiol reagents and abrogation of this effect by dithiothreitol and cysteine, they were identified as cysteine proteinases. Proteinase I (Mr= 95,000) and proteinase II (Mr= 35,000) were active against the β-chain of insulin releasing characteristic fragments. However, differences in substrate specificities of the two enzymes could be observed by using synthetic peptides that represent sequences 1–6, 8–18, and 20–30 of the insulin β-chain. Furthermore, the synthetic tetrapeptides Arg-Gly-Phe-Phe, Arg-Gly-Leu-Hyp, and Arg-Arg-Phe-Phe were hydrolyzed by the two proteinases releasing Phe-Phe and Leu-Hyp, respectively. Compared with Arg-Gly-Phe-Phe, the rates of hydrolysis of Arg-Gly-Leu-Hyp and Arg-Arg-Phe-Phe at substrate concentrations of 1 mM were 91% and 63% (proteinase I) and 80% and 57% (proteinase II), respectively.

Notes: Three-dimensional reconstruction of a binucleate intermediate precyst of Pneumocystis carinii was performed from serial-thin sections using the CATIA (Conception Assistée Tridimensionnelle Inter Active) Dassault system program. The presence of a mitochondrion, complex well-developed endoplasmic structures, and numerous Golgi vesicles was established. A better understanding of the ultrastructure of rabbit-derived P. carinii stages made it possible to formulate hypotheses on the evolution and physiology of the endomembrane system. Thus, the presence of the well-developed endoplasmic saccular structure and more than 230 Golgi vesicles in its vicinity might be implicated in the differentiation of the parasite surface structures and might also be related to nuclear division and individualization of intracystic bodies.

Notes: Brusca, R. C. & Brusca, G. J. 1990. Invertebrates. Sinauer, Sunderland, MassachusettsMiller, H. R. P. (ed.) 1990. Immunity to and Diagnosis of Internal Parasitism. Vol. 9, no. 2. Revue Scientifiqueet Technique, Ofie International Des Epizootics. Office International Des Epizootics, Paris, France.Preston, T. M., King, C. A. & Hyams, J. S. 1990. The Cytoskeleton and Cell Motility. Chapman and Hall, New York, New York.

Notes: Reinvestigation of the type population of the sorocarp-forming ciliate Sorogena stoianovitchae Bradbury & Olive, 1980 using the Fernández-Galiano technique and various electron-microscopy techniques (scanning electron microscopy, freeze-fracture and ultrathin sections) expands the observations reported in the original description of the species, Sorogena stoianovitchae is a colpodid ciliate with oral ciliature consisting of 25 ciliated paroral dikinetids on the right and 3-5 small adoral organelles on the left of an elongated and domed oral slit, resembling that of the genus Platyophrya. Sorogena stoianovitchae divides in the free swimming condition and not in a division cyst, as is the case in the colpodids sensu stricto (s. str.), e.g. Colpoda, Bresslaua. or Tillina, As shown in a detailed light-microscopy study, morphogenesis in S. stoianovitchae is of the stomatic mode typical for certain colpodid ciliates. Based on the wealth of new information the phylogenetic position of S. stoianovitchae is discussed at some length and arguments are given in favor of the following classifications: S. stoianovitchae Bradbury & Olive, 1980 currently sole member of the family Sorogenidae Bradbury & Olive, 1980; order Sorogenida Foissner, 1985;subclassColpodiaFoissner, 1985; class Colpodea Small & Lynn, 1981. This investigation facilitates the discovery of further members of this genus reported primarily from the tropical and subtropical zone.

Notes: Sorogena stoianovitchae is an unusual ciliated protozoan with a life cycle characterized by the aggregation of individual trophic cells to form a multicellular sorogen that rises from the liquid culture medium surface by the secretion of a stalk. The noncellular stalk is a tapered, longitudinally furrowed structure composed of a fibrillar matrix that is initially hydrated, but with time dehydrates, the stalk becoming thin and brittle. This dehydration is of importance from the earliest stages of stalk formation since it results in the formation of the outer sheath-like region of the stalk that appears to provide much of the support of the stalk. Cytochemical tests of the stalk for polysaccharides (including acidic mucopolysaccharides) and proteins are positive. Proteolytic enzymes degrade the stalk. Lectins specific for glucose and N-acetyl-D-glucosamine bind to the stalk. Gas chromatography analysis detected the presence of fucose, glucose, glucosamine, and arabinose, as well as a variety of amino acids, predominantly glycine. The cytochemical and biochemical tests, the ultrastructural data, and the behavior of the stalk material suggest that the staik is composed of a matrix of complex protein-polysaccharide molecules.

Notes: Nosema disstriae, a parasite of the forest tent caterpillar, Malacosoma disstria, was cultured with cell lines UMN-MDH-1 (Malacosoma disstria), IPLB-1075 (Heliothis zea), and BTC-32 (Triatoma infestans). Infected cultured cells were used to infect the healthy cell lines. Electron micrographs of thin sections of 6-day-old cultures revealed infected cells that exocytosed vesicles containing vegetative and immature sporulating forms of the parasite. Some of these forms were believed to be responsible for intercellular transmission of the parasite. The spread of infection was augmented by culturing the cells at high densities; if the density was too low, there was little or no cross infection. Cross infection was inhibited, but not blocked completely, by high osmolality of the culture medium. The yield of spores from a confluent cell monolayer at the end of growth was generally 1–4 × 107 per ml of culture medium.

Notes: The fine structure of the trophozoite of Acanthamoeba palestinensis with a special emphasis on the Golgi complex, microbodies, and mitochondria has been examined. Golgi complexes are distributed throughout the cytoplasm but are most abundant in the perinuclear region. Usually two Goigi complexes are found in the same plane on opposite sides of the nucleus. One of them appears to be in an intimate association with the nuclear membrane. The region of contact contains compact cisternae, vesicles of various sizes, as well as granular and amorphous electron-dense material. Structural changes in the nuclear envelope are also observed in this area. A structure consisting of a Golgi complex and electron-dense microtubule organizing center, comparable to the centrosphere of other Acanthamoeba species, has been observed. Microbodies, surrounded by a single unit membrane and containing a granular matrix and tubular inclusions, are scattered throughout the cytoplasm. These organelles, circular (∼1 μm in diameter) or ovoidal (∼1 μm in length and ∼0.5 μm in width) in section, have often an irregular outline. These microbodies are probably the morphological equivalent of peroxisomes and glyoxysomes. Most mitochondria show a typical structure including tubular cristae and intracristal inclusions. Occasionally mitochondria with two apposed double membranes running through the midline are found. Such atypical cristae have never been reported in small amoebae before.