ALBERT

Notes: A philasterine scuticociiiate 82-133 μm in length and 29-51 μm in width is described from the intestine of the solitary tunicate Ascidia ceratodes. It has 44 somatic kineties, an anterior dorsal bald spot, and a triangular scutico-vestige which lies above recessed posterior oral polykinetids.

Notes: Saudi Arabian camels {Camelus dromedarius) are infected with three species of Eimeria: E. dromedarii (28.4%), E. rajasthani 〈 22.2%), and E. cameli (19.2%); 41.6% of the animals examined were positive. The highest prevalence of infection was reported in the western region of the country. Mixed infection with two Eimeria species is most common; E. dromedarii was most frequently and generally the most predominant species. Eimeria dromedarii and E. rajasthani are described for the first time from Saudi Arabian camels.

Notes: Tetrahymena of the T. pyriformis complex collected from varied habitats in Malaysia, Thailand, and The People's Republic of China include strains of the micronucleate species T. americanis and T. canadensis and the amicronucleate T. Pyriformis and T. elliotti. Two new breeding species are described—T. malaccensis from Malaysia and T. asiatica from China and Thailand. Two wild selfers from China and some of the amicronucleate strains from all three countries fall into isozymic groups similar to named micronucleate and amicronucleate species. The T. patula complex is represented by two groups of clones from Malaysia that fit the morphological description of T. vorax. They, however, have radically different isozymic electrophoretic patterns and both groups differ from those of previously described T. vorax. As their molecules indicate relationships to other “T. vorax” strains as distant as that between T. vorax and T. leucophrys, they are considered to be new species, T. caudata and T. silvana. A third new breeding species, T. nanneyi, was identified among strains previously collected in North America. Viable immature progeny were obtained from the new strains of the five breeding species. Maximum temperature tolerances were determined for the new strains of four of the breeding species.

Notes: The cellular bases of the powerful cytolytic activity of the human protozoan parasite Entamoeba histolytica were explored by studying the effect of the virulent strain HM1:IMSS on epithelial monolayers of MDCK cells using a combination of time-lapse microcinematography and transmission and scanning electron microscopy. Early alterations of the epithelial cell membranes were detected by measuring changes in the transepithelial electrical resistance of MOCK monolayers mounted in Ussing chambers. The aggressive mechanism of E. histolytica trophozoites was found to be a complex, multifactorial phenomenon that included hit-and-run damage to the plasma membrane of effector cells mediated through contact, phagocytosis of lysed or apparently intact, but detached, MDCK cells, and inlracellular degradation of ingested cells. Following contact with amebas, the epithelial monolayers showed a pronounced lowering of transepithelial resistance, opening of tight junctions, distortion of microvilli, surface blebbing, and the presence of minute focal discontinuities in the plasma membrane. There was no evidence of amebic exocytosis, membrane fusion, or junction formation between the parasite and host plasma membranes. Although modifications in the epithelial cell membranes usually preceded lysis, the cytolytic activity of the parasite did not exclusively involve damage to the plasma membrane of the cultured host cells but also was mediated by avid phagocytosis, the displacement and separation of neighboring cells by means of pseudopodial activity, and the “pinching-off” of the peripheral cytoplasm of epithelial cells.

Notes: Five new species of myxosporean parasite are described from cultured tilapias in Israel. These are: Myxosoma sarigi. Myxosoma equatorialis, Myxobolus israelensis. Myxobolus agolus, and Myxobolus galilaeus. The first four were found in hybrids of Oreochromis aureus x Oreochromis niloticus while Myxobolus galilaeus was found in Sarolherodon galilaeus. In addition, M. sarigi. M. israelensis, and Myxobolus sp. were also found in S. galilaeus. In the light of the present study, the taxonomy of myxosporean infections in tilapias is modified. Mature spores may localize in the melano-macrophage centers of the spleen and kidney where ihey may eventually be destroyed. Nc cases of mortality have so far been associated with these parasites.

Notes: Oocysts of Caryospora cobrae n. sp. were isolated from an Indian cobra, Naja naja Linnaeus from West Bengal, India. The sporulated oocysts were spherical to subspherical, 16.5-19.5 times 16.5-18.0 μm, and had a micropyle but lacked a polar granule and oocysl residuum. The sporocysts were piriform, measuring 12.0-16.5 times 9.0−12.8 μm; a Stieda body and sporocyst residuum were present.

Notes: World Health Organization. 1984. The Leishmaniases.

Notes: After a thin membranous envelope surrounding the cell body and cilia of Colpoda steinii has been formed, the main mass of the proteinaceous cyst wall is deposited without exocytosis. It can be composed of two layers, the denser and wrinkled ectocyst and the smooth-walled endocyst; however, the ectocyst may be missing. Evidence is presented that ecto- and endocyst are formed from vesicles derived from abundant rough endoplasmic reticulum which appears at the time of wall formation. The cilia are retained and become embedded in the peripheral cytoplasm. Synthesis of RNA and protein is required as actinomycin C and cycloheximide block cyst formation. Calcium is required during a sensitive phase prior to encystment.

Notes: Trypanosoma brucei strain 366D trypomastigotes grown at 37°C in the presence of a human fibroblast cell line formed foci underneath the feeder cells whereas trypanosomes grown in the presence of a human epithelial cell line grew only in the culture supernatant. A culture system was developed to study the differentiation of bloodstream trypomastigotes grown in the epithelial cell system into procyclic trypomastigotes at 27°C. The morphological differentiation into the procyclic form was complete by 48 h. Cell division did not occur until 30–40 h after transfer to 27°C. Various characteristics of this system were examined, including the effect of the feeder layer, the type of medium, the presence of the metabolites cis-aconitate and citrate, the preadaptation period, and the trypanosome cell concentration. The respiration of the recently differentiated procyclic cells was less sensitive to inhibition by CN-than that of established procyclic forms, implying a delayed appearance of complete mitochondrial oxidative pathways. This trypanosome differentiation system has the advantage that the animal host is not needed and the entire process is carried out in in vitro culture.

Notes: The flagellated protozoan, Giardia agilis, was isolated from tadpole small intestine and examined by scanning electron microscopy and interference reflexion microscopy. The general morphology of the G. agilis trophozoite is similar to G. muris and G. duodenalis, but with modifications that reflect its elongated form. Interference reflexion microscopic analysis of attachment of G. agilis reveals a pattern of focal contacts by the lateral crest of the ventral disc, the ventrolateral flange, the lateral shield, and by numerous microvillus-like appendages found along the lateral border of the trophozoite. The pattern of focal contacts was observed to be dynamic; trophozoites were observed to make and break the focal contacts in a relatively short time and to glide along the surface of the substratum without breaking focal contacts.

Notes: The fine structure of stages in mitosis in a colorless euglenoid, Anisonema sp., reveals that chromosomes remain condensed throughout the life cycle and are attached to the nuclear envelope at interphase. The onset of mitosis is marked by the anterior migration of the nucleus towards the base of the reservoir and by elongation of the nucleolus. The nuclear envelope persists throughout mitosis. Microtubules are generated in the peripheral nucleoplasm adjacent to the envelope and attach to the chromosomes while they are still associated with the envelope. The region of microtubular contact develops into a distinct layered kinetochore as the developing spindle with attached chromosomes separates from the nuclear envelope and moves into the nucleoplasm. The mature spindle consists of a number of subspindles each containing about 8–10 microtubules and a few associated chromosomes. Both chromosomal and non-chromosomal microtubules are present in each subspindle and extend towards the envelope terminating at or near the nuclear pores. Chromosomal segregation is concomitant with nuclear elongation. By late division, an interzonal spindle develops in the dumbbell-shaped nucleus and nucleolar separation occurs. Continued invagination of the nuclear envelope in the region of the interzonal spindle eventually separates the daughter nuclei. A remnant of the interzonal spindle persists in the cytoplasm until cytokinesis.

Notes: The fine structure of exoerythrocytic merogony of Plasmodium berghei was studied after perfusion-fixation of rat livers from 51 h post-inoculation onwards. Meroblast formation was effected by clefts originating from the parasite plasmalemma and by fusion of vacuoles with each other. Invaginations at the periphery resulted in labyrinthine structures providing the parasites with an enormous increase in surface area, which might facilitate exchange of metabolites. When the parasitophorous vacuole membrane collapsed, the newly formed merozoites were lying free in the hepatocytic cytoplasm, which degenerated until the merozoites were sticking together by a stroma, obviously a remnant of the host hepatocyte. Groups of merozoites, still kept together by the spongy stroma, were subsequently released in the bloodstream. At 53 h most of the developmental stages leading to the release of merozoites could be found and thereafter parasite numbers decreased while large granulomas became apparent.

Notes: This study examines previously undescribed general and cytopharyngeal features of the genus Entodinium. The cytopharynx contains three types of microtubular ribbons underlying the cytostomal membrane as well as a loose palisade of nematodesmata. A protoesophagus composed of microtubular bundles associated with a fibrous wall lies internally to one side of an extrusible peristome on which the adoral zone of syncilia (AZS) is mounted. Macronuclear structures are very similar to those of other ophryoscolecids. The micronucleus has chromatin bodies forming a compact mass but lacks the thick wall found in other species. A tubular spongiome surrounds the contractile vacuole and the cytoproct is relatively undifferentiated. Cortical structure follows the usual five-layered ophryoscolecid pattern with subcortical barren kinetosomes arranged into indistinct kineties. The infraciliature of the AZS has kinetosomes set upon a subkinetal rod and with associated bifurcated kinetodesmata and transverse microtubules, some of which extend into the cytopharynx. Components newly described for Entodinium are the one to three postciliary microtubules and the interkinetosomal centro-lateral strand, all of which are present in other species of ophryoscolecid ciliates. The infraciliature of the paralabial ciliary tuft shows similar components to that of the main AZS, but lacks the subkinetal rod. The microtubular components of the cytopharynx are discussed in relation to the “alimentary” structures in other ophryoscolecids, and a relationship of these structures to dietary differences is suggested.

Notes: Cryptosporidium parvum oocysts isolated from calf feces were examined by scanning electron microscopy during excystation. Intact C. parvum oocysts were spheroid to ellipsoid, ≅3.5 × 4.0 μm, with length: width ratio = 1.17. The oocyst wall had a single suture at one pole, which spanned 1/3 to 1/2 the circumference of the oocyst. During excystation the suture dissolved, resulting in a slit-like opening, which the sporozoites used to exit the oocyst. Sporozoites were 3.8 times 0.6 μm and had a rough outer surface.

Notes: Special ultrastructural characteristics of the haptorid soil ciliate Enchelydium polynucleatum Foissner, 1984 are the restriction of the parasomal sacs to the area of the “brush” and finger-like projections of the food vacuole membrane into the lumen of the vacuole. The general organization of the infraciliature is similar to that of Spathidium and some buetschliids because the anterior ends of the somatic kineties are condensed and obliquely bent. Enchelydium is similar to haptorids and buetschliids in possessing monokinetid somatic fibrillar structures with the classical fibrillar associates: 1) a short kinetodesmal fiber; 2) two transverse microtubular ribbons; 3) a long postciliary microtubular ribbon; and 4) a system of overlapping subkinetal microtubules, which seems to be absent in the buetschliids. Unlike Spathidium and all other haptorids so far investigated ultrastructurally, serial sections show that there are no oral dikinetids, as in the endocommensal buetschliids and balantidiids. Instead, three to six anterior kinetids in each ciliary row have nematodesmal bundles extending into the cytoplasm and surrounding the cytopharynx. These kinetids lack cilia and all fibrillar associates except enlarged transverse ribbons, which extend anteriorly and inwards to support the cytopharynx. Other similarities between the buetschliids and Enchelydium are the conspicuous rough endoplasmic reticulum and abundant sausage-like vesicles in the oral region. As in other haptorids, Enchelydium has two types of toxicysts and one type of mucocyst. These observations strongly suggest that Enchelydium belongs to the ancestral stock of both the Haptorida and the Archistomatida. The similarities in the somatic and oral infraciliature and ultrastructure of the Haptorida and the Archistomatida suggest that they belong to the same subclass, Haptoria Corliss, 1974.

Notes: Resting cysts and the excystment process in the freshwater ciliate Euplotes muscicola were studied by both light and scanning electron microscopy. Groups of distinctly crested resting cysts adhere to the substrate. Silver-stained preparations reveal surface conservation of dorsal kinetosomes and dorsal argyrome while ventral organelles are directed inward. Excystment involves the development of an expanding excystment vacuole concurrent with a localized thinning on the dorsal cyst wall surface. Cells exit through the pre-formed ostiole, mid-dorsal region first, initially by the force of cytoplasmic streaming, but later aided by cirral movement. Newly emerged cells retain the excystment vacuole and show no dorsal ridging. As the cell expels its excystment vacuole and partially unfolds, normal trophont morphology is re-established. Both cyst structure and cyst typology have implications for hypotrich taxonomy.

Notes: The cytosol fraction of a symbiont-bearing strain of Amoeba proteus exerted a lethal effect when injected into symbiont-free amoebae of the original strain. The lethal factor appeared to be a protein with a molecular weight of over 200,000. While the effect of the lethal factor on the nucleus was reversible, the host cytoplasm was permanently damaged so that it could not form a viable cell when combined with a normal nucleus.

Notes: Laybourn-Parry, Johanna 1984. A Functional Biology of Free-Living Protozoa.

Notes: Seven monoclonal antibodies specific for mammalian β-tubulin demonstrate the microtubule cytoskeleton of Toxoplasma gondii and Leishmania donovani by indirect immunofluorescence microscopy. Immunoblots of T. gondii and L. donovani proteins separated by SDS polyacrylamide gel electrophoresis confirm the specificity of the monoclonal antibodies for tubulin. Differential staining of flagellar and subpellicular microtubule populations was not seen in L. donovani with these antibodies. All seven antibodies also detected the subpellicular microtubules of T. gondii, but the polar ring and conoid of this organism was not visualized by any of them. This technique provides a rapid and specific way to assess microtubular organization in whole organisms.

Notes: Lee, J. J., Hutner, S. H. & Bovee, E. C., eds. 1985. An Illustrated Guide to the Protozoa.

Notes: .Attention, perhaps overdue, is drawn to the extent and significance of endosymbionts (xenosomes sensu lato) in the cytoplasm and nuclei of many protozoa from diverse taxonomic groups. Even more importantly, recent advances in the study of such intimate associations are reviewed and discussed and their impact on broader problems of cell biology and evolution are stressed. Workers inside and especially outside the fields of protozoology and parasitology have often neglected such data, failing to appreciate their relevance to significant problems in their own fields of investigation. The major topics covered by speakers in the Symposium (to which this paper serves only as an introduction) include the following, in order of their presentation: terminology for the symbiont-host relationship and a brief overview of the field; the evolutionary problem of the origin of contemporary associations, including cell organelles such as mitochondria and plastids; the adaptive value of endosymbionts to their protozoan hosts; mechanisms of establishment, maintenance, and integration of such foreign bodies/invaders in their unicellular eukaryotic host cells; and the extent of algal and bacterial endosymbioses in diverse protozoan groups. In all papers, the principal relatively well studied complexes used as examples are the following: various kinds of algae in the larger foraminifera and in ciliates, radiolarians, and acantharians; the several types of bacteria in the cytoplasm of Amoeba and of Pelomyxa; the endonuclear bacterial symbionts of Paramecium; the cytoplasmic prokaryotes in Paramecium and in Parauronema; and the methanogenic bacteria of certain ciliates.

Notes: .It is generally accepted that in symbiotic systems involving algal species as cellular endobionts there is some positive benefit to the host organisms. In this paper special consideration is given to the larger foraminifera, protozoa that serve as very useful model systems for the study of aspects of inter/intracellular integration and adaptation—living, as they do, in nutrient-limited but well illuminated shallow tropical seas and containing endosymbiotic algae in abundance. A considerable amount of information is now available on physiological as well as morphological adaptations of the host species to pigmented protists representing diverse algal divisions (phyla). Brief mention is also made of bacterial endosymbionts of certain ciliates.

Notes: .In this paper the concept of “xenosome” is greatly expanded from its current usage, which has been based on its application during the past 10 years by Soldo and co-workers solely to certain bacterial invaders of the cytoplasm in species of a single genus of marine scuticociliales. The author proposes that the term now be considered to embrace all DNA-containing, membrane-bounded bodies or organelles—prokaryotic or eukaryotic in original nature—found within the cytoplasm or nucleus of eukaryotic cells of any or all kinds, whether the occupation (“colonization”) is temporary and transient or permanent and stable. Thus, virulent or pathogenically infectious organisms can be included as well as the commonly recognized cell endosymbionts sensu stricto, which are often mutualistic in nature. Of significance, such “normal” cell organelles as plastids, mitochondria, and even nuclei may also be embraced by this expanded definition of xenosome, based on the conjecture that these inclusions might have been “alien” or “foreign” extracellular, independent, free-living organisms in their own past evolutionary histories. The author's enlarged concept and unifying principle allows more meaningful comparative consideration of the numerous and diverse kinds of xenosome-host interrelationships, many of which involve species of protozoa and algae from a large number of the taxonomic groups comprising the kingdom Protista.

Notes: Protozoa are now being recognized as important members of planktonic food webs. This is due to the inclusion of microbial links in our paradigm of trophic relationships. Heterotrophic microflagellates and ciliates are major grazers of bacteria. They can stimulate production through nutrient recycling and can transform microbial production into larger particles, which are then available for macroconsumers. In this paper we add new groups, the small (〈 20 μm) ciliates and myxotrophic flagellates, to the planktonic food web.

Notes: .Important progress has been made in recent years towards better understanding of the establishment and maintenance of endosymbiosis in protozoa and of the eventual integrative mechanisms involved. Still, many problems remain to be investigated more thoroughly. In this paper, while treating and reviewing the subject broadly, particular and more detailed attention is given to three selected systems: endonuclear symbiosis by Holospora bacteria in Paramecium; algal (Chlorella) relationships with the “green”Paramecium (P. bursaria) as host; and the rod-shaped bacteria found in the cytoplasm of Amoeba proteus. Data concerning the physiology of food vacuoles, membrane transport, photobehavior, recognition specificity, enzyme activity, and the like are presented and reviewed and discussed in light of the growing literature on the overall subject of “endocytobiology.” Emphasized is the complicated network of interactions between symbiotic partners and the importance of the development of integrative mechanisms in the evolution of the many intimate associations known at the cellular level today.

Notes: Long neglected has been the extensive and more or less intimate association of protozoa with a wide variety of other cells, either prokaryotic or eukaryotic in nature. Yet study of such relationships can provide important information concerning certain basic aspects of cellular evolution in general. A survey is offered here of the whole range of such symbiotic associations (i.e. with species of protozoa serving as hosts) with the purposes of drawing attention to the exciting possibilities of such research and of reviewing significant findings made to date. Because of the vastness of the overall field, examples and discussion are primarily limited to consideration of the following major studies: methanogenic bacteria in certain ciliates, bacterial endosymbionts of the large freshwater amoeba Pelomyxa palustris (itself an amazing organism from an evolutionary/phylogenetic point of view), the rod-shaped bacteria found in Amoeba proteus, the “Greek-letter” prokaryotes of Paramecium species, the xenosomes (sensu stricto) of the marine scuticociliate Parauronema acutum, and the diverse algal endosymbionts of similarly diverse protozoan taxa–ciliates, flagellates, radiolarians, acantharians, and foraminifera.

Notes: Soil is the focus of organic matter turnover in terrestrial ecosystems and is an interstitial mosaic of microsites composed of particle aggregates and pore spaces, where transformation, decomposition, mineralization, and humification of organic matter takes place. Microorganisms and animals are scattered discontinuously in these microsites. Microarthropods and larger fauna increase the rate and amount of mineralization by comminution of organic matter and by redistribution of microsites through movements of earthworms and large arthropods; however, mineralization and return of nutrients to plants occurs in the community of bacteria, fungi, protozoa, and nematodes living in the water films covering aggregates and filling pore spaces. Protozoa, especially small amoebae, are important bacterial grazers because they can enter tiny spaces unavailable to nematodes. The latter graze bacteria, fungi, and protozoa. Protozoan and nematode predation increase the amounts of soluble nutrients and decrease the competitive abilities of bacteria, thus making these nutrients more available to plants. Protozoa enhance nutrient recycling out of proportion to their biomass.

Notes: Following conjugation of the hypotrichous ciliate Euplotes aediculatus, the posterior fragments of the old (prezygotic) macronucleus persist until after the first vegetative division. These fragments remain viable during exconjugant development as shown by their ability to regenerate should the cell's new macronucleus be damaged. It thus seemed possible that these parental nuclear fragments might participate in the development of the new macronucleus and/or the crucial post-conjugant cortical reorganization that restores the exconjugant cell's ability to feed. This idea was tested by damaging the posterior fragments with various doses of microbeam ultraviolet (UV) light and assessing the results of such treatment on subsequent cortical and nuclear development. When the posterior fragments of the macronucleus were irradiated at the beginning of cortical morphogenesis, the new macronucleus in 1/3 to 1/2 of the cells assumed a “folded” appearance but did not mature. These cells did not undergo cortical reorganization. Cells irradiated at earlier stages did not detectably develop an oral apparatus; their new macronucleus remained arrested at the spherical anlage stage. The results show that the posterior fragments of the parental macronucleus are necessary for normal nuclear and cortical development. These old nuclear fragments appear to influence the growing macronuclear anlage directly and probably the cortex as well. There also appears to be an information flow from the non-irradiated partner of a persistently joined exconjugant doublet to its irradiated counterpart, enabling normal anlage and cortex development in the irradiated cell.

Notes: Envelope development in Trachelomonas lefevrei (Deflandre) begins with the production of short, coarse mucilaginous strands, morphologically similar to the mucilage produced by non-enveloped euglenoids. These initial mucilaginous secretions subsequently become impregnated with manganese (Mn) and/or iron (Fe). Continued mucilage secretion and mineralization results in a mature envelope that is characteristic for the species. When these mature envelopes are treated with oxalic acid to remove the Mn and Fe, the envelopes collapse and are composed only of short, coarse mucilaginous strands similar to those present during early stages of development, prior to their mineralization. Brief treatment with 10 mM EDTA renders dark envelopes colorless, and our EM-EDS analyses show that this corresponds to a loss of Mn from the envelope; however, if Fe is present in the envelope, it is unaffected by the brief treatment. The mucilage present during early stages of envelope development and that remaining after complete demineralization is also morphologically similar to the mucilage in the plug at the anterior end of the envelope.

Notes: Two new species of the genus Nosema (Microsporida: Nosematidae) are described from the Mexican bean beetle, Epilachna varivestis (Coleoptera: Coccinellidae) and their life cycle stages studied by light and electron microscopy. Both species are monomorphic and disporous: they develop in direct contact with the cytoplasm of host cells and the nuclei of all stages are diplokaryotic. The more virulent species produces systemic infections most extensively in the adipose tissue, muscles, and Malpighian tubules of larvae and also invades the reproductive tissues of adult beetles. During merogonic development, it forms chains of diplokaryotic meronts. The fine structure of the sporoblast nuclei shows clumped material in the pole of each nucleus opposite their common plane of apposition. Spores are straight to slightly curved and ovocylindrical in shape and they measure 5.3 ± 0.13 × 2.1 ± 0.03 μm. The less virulent species also invades most host tissues but does not develop in the midgut epithelium; the Malpighian tubules are the principal site of its development and it also invades the ovaries and testes of adult beetles. Merogony occurs exclusively as the result of binary fission of diplokaryotic meronts. The plasmalemma of the meronts is covered with a thin deposit of exospore material upon which are located closely packed tubules that encircle the body transversely. A thickened deposit of exospore material on the surface of the diplokaryotic sporonts later obscures these tubules. Other tubules occur free in the host cell cytoplasm or attached to the plasmalemma of meronts and sporonts. Secretory granules also occur free or in chains in the host cytoplasm and are probably produced from the surface of the sporoblasts. Sporoblasts also contain an unusual cup-shaped organelle associated with a dense body, which is apparently involved in the formation of the polar tube and its associated organelles in the anterior part of the spore. Spores are ellipsoidal to slightly pyriform and measure 4.7 ± 0.06 × 2.6 ± 0.03 μm.

Notes: Marshall, K. C., ed. 1984. Advances in Microbial Ecology.

Notes: Starved Tetrahymena thermophila (or cells growing in Holz's defined medium) are attracted by a chemosensory response to complex peptide mixtures as proteose peptone, yeast extract, and extracts of blood platelets containing platelet-derived growth factor. Starved cells are also significantly attracted by mixtures of amino acids and of nucleosides of Holz's defined medium; however, no individual amino acid or nucleoside could be identified as the major chemo-attractant.The positive chemosensory response can be abolished by cycloheximide (CHX) but is insensitive to actinomycin D and puromycin, possibly indicating that de novo RNA and protein synthesis are not necessary for a change in chemosensory behavior. Three mutants resistant to CHX show normal response in the presence of the drug.The possible role of peptides as naturally occurring food signals of Tetrahymena is discussed.

Notes: The premeiotic micronucleus of Tetrahymena thermophila elongates parallel to the long axis of the cell. In fixed cells one end of this crescent micronucleus appears thicker than the other, and either end may be oriented toward the anterior of the cell. Three families of repeated DNA sequences have been localized in the crescent micronucleus by in situ hybridization. Two micronucleus-specific sequences hybridize all along the crescent, but preferentially toward the ends. A macronucleus-retained sequence hybridizes preferentially to the half of the micronucleus at the thick end. Thus the arrangement of DNA sequences in the crescent micronucleus is nonrandom.

Notes: Simple culture conditions that allow good growth and high yields of trypomastigotes are described. The proportion of metacyclic trypomastigotes increases with the concentration of hemin in the culture medium, reaching a peak of 80% after 10 days with 20 mg hemin/liter.

Notes: Thuricola folliculata is a sessiline, loricate peritrich ciliate. Its somatic pellicle consists of annular transverse crests and includes the plasma membrane, the alveolus, the electron-dense epiplasm, and the subepiplasmic layer. Cytokinesis occurs along anoral-aboral median plane where two fissures develop, one at the oral and one at the aboral end of the peritrich. Each fissure results from simultaneous formation of two furrows on opposite sides of the cell. At the end of cytokinesis, both trophozoites remain joinedto each other by an intercellular bridge. During cytokinesis, microfilament bundles appear at the level of the subepiplasmic layer in the fission plane; they are distributed in two arcs, one oral and one aboral, and may be responsible for the formation of the four furrows.The cross-sectioned microfilament arc is 1 pm wide and about 0.1-0.2 pm thick at first and later can be more than 1 pm in diameter; it shows many microfilaments, 3-10 nm in diameter and oriented parallel to the fission plane, and also many dense corpuscles 25-55nm in diameter. Then both arcs join each other to form a microfilament ring. This ring is delimited by discontinuous dense borders and a boundary layer. The microfilament ring seems structurally analogous to the contractile ring of various dividing cells, where itworks like a sphincter. The dense corpuscles, the discontinuous dense borders, and the boundary layer of T. folliculata have not been reported in any other ciliates.

Notes: The sequence of morphologic events associated with Ichthyophthirius rnultifliis invasion of gill epithelium began in the theront with differentiation of secretory mucocysts and the perforatorium. After escaping from the cyst the theront, which stained intensely with Mallory' stain, sought a host. As it approached the host epithelium, the contents of the mucocysts were discharged, enveloping the ciliate in sticky material, which made initial contact with the host epithelium. Rapid penetration by the theront caused disruption of one or more host cells and resulted in a focal necrosis associated with the anterior margin of the ciliate. Within five minutes postexposure, the parasite completed its invasion of the epithelial layer and stained less intensely. The remnants of host cells disrupted during its entry surrounded the trophont until they were ingested by the parasite. Within 40 min postexposure, synthetic activity of the parasite appeared to increase as evidenced by an abundance of organelles, particularly ribosomes and crystalline mucocysts. At this point, the overlying host epithelium appeared normal.

Notes: This study examines the relationship between phagosome acidification and phagosome-lysosome fusion events using phagocytized Glugea hertwigi spores. The incidence of lysosome fusion with Glugea spores in phagosomes of mouse peritoneal macrophages and of Tetrahymena was monitored using colloidal gold and acridine orange as labels for secondary lysosomes. Over 80% of the Glugea phagosomes remained segregated from the labeled compartments in macrophages after 60 min; this inhibition of fusion was still evident after 4 h. In Tetrahymena, Glugea spores also showed a high capacity to block fusion with secondary lysosomes (67%); however, spores coated with cationized ferritin showed an 80% fusion rate with labeled acidic compartments (i.e. lysosomes) after 60 min with both Tetrahymena and macrophages. The pH of phagosome compartments was monitored by measuring the emissions of fluorescein isothiocyanate (FITQ-labeled Glugea ingested by Tetrahymena. Tetrahymena phagosomes with FITC-Glugea did not acidify within the first hour after phagocytosis; however, phagosomes with cationized ferritin-labeled Glugea underwent acidification during this time period. This acidification took place although the capability of the host cells' lysosomes to fuse was blocked by pretreatment with poly-D-glutamic acid. The cationized ferritin bound to Glugea spores was uncoupled from the spore wall prior to fusion with colloidal gold-labeled compartments. In vitro testing showed that ferritin dissociation requires an acid pH, indicating that phagosomes acidify prior to lysosome fusion.

Notes: Incubation of radioactively labeled parasitized (Plasmodium berghei) erythrocytes (PE) with adherent peritoneal exudate cells in the presence of 10% (v/v) fresh mouse serum (NMS) resulted in the uptake of a proportion of radioactive material (PE). Inactivation of the added serum by heat or zymosan treatment resulted in diminished uptake of radioactivity. These results suggest that PE activated complement. Incubation of fresh NMS with PE reduced the hemolytic complement level of the serum as shown by its subsequent decreased ability to lyse antibody-coated rabbit red blood cells. No such effect was found when uninfected erythrocytes from either infected or uninfected blood were preincubated with fresh NMS. Thus, PE or PE-derived material activated complement. Addition of EGTA during incubation of fresh NMS with PE did not inhibit the decrease in complement level. This indicated that complement was activated by the alternative pathway. Complement levels decreased even when fresh NMS and PE were incubated in the presence of EDTA (which inhibits both classical and alternative pathway activation), suggesting that a complement activating factor (or a complement inhibitor) was released from the PE. However, lysis of PE after incubation with either fresh rabbit or guinea pig serum did not occur unless anti-mouse erythrocyte antibody was added. The production of a complement-activating factor by PE might explain part of the decreasing complement levels during infection and might enable the parasite to escape from a complement-mediated defense mechanism of the host.

Notes: The reaction of the contractile vacuole of Amoeba proteus to single and multiple phagocytosis under controlled conditions has been studied. Fluid intake into the cytoplasm from the phagosomes induces secretion by the contractile vacuole of equivalent excess volumes:. Vacuolar response is rapid (200 sec) and may be initiated by increases of protoplasmic hydration of as little as 1%. Cytoplasmic uptake of fluid from the phagosome can occur against an osmotic gradient; thus some form of active transport is implied.

Notes: When amebae were incubated with latex beads, cyanide-insensitive oxygen consumption increased nearly two-fold. This cyanide-insensitive respiration was inhibited by salicylhydroxamate. Furthermore, cell fractionation studies revealed a localization for a portion of the NAD(P)H oxidase activity in phagolysosomes. The presence of low concentrations of divalent metal during fractionation resulted in an increased yield of oxidative activity in the phagolysosome fraction. In addition, the phagolysosome membrane was enriched about two-fold in a b-type cytochrome. These results show that oxidative metabolism in amebae has some striking similarities to the respiratory burst oxidase of neutrophils.

Notes: C57B1/6 mice were infected with Brasil strain Trypanosoma cruzi trypomastigotes. The leg muscles of the mice were serial-sectioned with a cryostat, and individual fibers were classified histochemically as type I or type II on the basis of succinic dehydrogenase or adenosine triphosphatase activity. Although markedly more type II fibers were present in the leg muscles, the percentage of infected type I fibers was nearly five-fold higher than type II. This is the first demonstration of a preferential in vivo distribution of T. cruzi in muscle fibers based upon muscle type.

Notes: A micropore membrane procedure to assay taxis by Entamoeba histolytica is described and the results of studies of responses to a variety of soluble substances, bacteria, an rat colon washings using this procedure are reported. Trophozoites migrated in blind well chambers through 8-m̈m pore size polycarbonate membranes but not nitrocellulose membranes up to 12 m̈m pore size. Amoebae were attracted toward fresh axenic culture medium (TYI-S), an enzymatic hydrolysate of casein (Trypticase), and a partially purified preparation of N-acetylneuraminic acid from egg mucin, but not purified N-acetylneuraminateora variety of other low molecular weight metabolites. The response was verified as chemotaxis by checkerboard analysis. Amoebae migrated most dramatically toward suspensions of all of seven bacterial species tested, including motile and non-motile, gram-negative and gram-positive rods and cocci. This response was diminished when the bacteria concentration gradient was eliminated. The response to bacteria culture filtrates was less than 10% of that to bacterial suspensions. A response to clarified washings from the rat colon was detected; this was diminished but not eliminated by filter sterilization of the washings. We concluded that some soluble molecules, possibly of intermediate molecular size, whole bacteria, and both soluble and paniculate components of the rat colon provide tactic stimuli for E. histolytica. Scanning electron micrographs of trophozoites migrating towards attractants through membranes showed narrow', extended pseudopodia entering the membrane pores, and enlarging spheres exiting as the cells proceeded through.

Notes: Extraction of whole promastigotes of Leishmania tropica major and L. donovani with a mixture of hexane and isopropanol (3:2) yielded three fractions containing immunological activity: 1) lipids, where the activity was determined by radioimmunoassay; 2) a lipopolysaccharide-like (LPS-like), water-soluble precipitate, where activity was determined both by radioimmunoassay and double gel diffusion, and 3) the phenol: water extract of the lipid-free promastigotes, where activity was followed by double gel diffusion.The use of a solid state, lipid-based radioimmunoassay for detection of leishmanial antigens provided a sensitive measure of their activity with a considerable degree of species and serotype specificity. We found antibodies to leishmanial lipids in sera from immunized rabbits, convalescent mice, and human patients with confirmed cases of cutaneous leishmaniasis or kala azar. There was very little activity in normal human or animal sera. Analysis by SDS-polyacrylamide gel electrophoresis of fractions from promastigotes surface-labeled with galactose oxidase and sodium borotritiate and preliminary immunochemical characterization of the LPS-like antigen showed that it contained galactose, but otherwise differed immunologically and chemically from excreted factor (EF), the best characterized leishmanial antigen.

Notes: In Isotricha prostoma, proteins of isolated cortices retaining the ecto-endoplasmic fibrillar boundary (EEB) have been separated by one- and two-dimensional gel electrophoresis. In addition to that of tubulin, four main bands corresponding to polypeptide molecular weights of 185, 35, 23, and 22 kd have been observed on 9% SDS-polyacrylamide gels. Supernumerary major bands within a low molecular weight range (11-19 kd) are evident after separation on 7.5-15% polyacrylamide gradient gels. Two-dimensional analysis discloses - 150 polypeptide spots, most of them focusing within an acidic pl range 4.6-5.3. The cortical fibrillar structures are still distinguishable after incubation in solutions containing 1.0 M K. Cl (12 h) or 0.6 M Kl (8 h). A prolonged KI treatment (52 h), however, results in the removal of EEB filaments (˜4 nm in diameter) and of a large part of the kinetid elements, so that the insoluble material is mainly represented by a reticulum of 6-nm filaments interconnecting the proximal regions of kinetosomes. Two proteins with MW of 58 and 63 kd appear to be constituents of this kinetosome-associated filamentous reticulum. Some of the cortical proteins with MW ≤23 kd are the most likely candidates for components of EEB filaments. The absence of decoration with heavy meromyosin confirms that actin is not a major component of Isotricha cortical cytoskeleton.

Notes: A semi-defined minimal medium, in which pantothenic acid is the only vitamin, was used to culture Plasmodium falciparum for the analysis of antimetabolite drugs. Analogs of riboflavin, nicotinamide, pyridoxine, and thiamin inhibited the growth of this parasite; for each drug, effects were much more pronounced after 96 h of exposure compared to 48 h. The most potent drug examined was 8-methylamino-8-desmethyl riboflavin (IC50 value approximately 1.0 times 10-10 M at 96 h). Avidin, a protein which complexes and thus inactivates biotin, did not affect parasite viability. Other antimalarial drugs, including chloroquine and quinine derivatives and antibiotics, were equipotent in the minimal medium and in RPMI 1640. Four strains of P. falciparum showed only minor differences in sensitivity to these antimetabolites. The use of prolonged drug exposure times and a vitamin-depleted medium allowed the preliminary characterization of antimalarial antimetabolites in vitro.

Notes: The complete intracellular cycle of five cloned stocks of Trypanosoma cruzi was quantified. Marked but stable interclonal differences were found in the length of the pre-replicative lag period (18.2-34.2 h), amastigote doubling lime (8.6-21.5 h), and duration of the complete intracellular cycle (96-215 h). Strong correlations were demonstrated between these characteristics as well as to the growth rate of the epimastigote stage of the same clones grown in liquid medium. These data demonstrate that the marked heterogeneity of the natural population of T. cruzi extends to the intracellular cycle of the parasite and has important implications for our understanding of Chagas’ disease.

Notes: An investigation was carried out to probe into the mating-type structure of a local population of the marine ciliate, Euplotes minuta. From this population, nine different mating types belonging to a unique set were isolated. The nine type-representative wild stocks analyzed were found to be heterozygous at the mating-type (mat) locus and provided, together with their sexual progeny, a total of 15 pure mating types. In E. minuta, the high-multiple nature of the basic mating system controlled by a series of peck-order alleles at a single locus should be considered a virtual certainty. The relationships among the genetic economies of the similar bottom-dwelling marine ciliates of the genus Euplotes, the E. vannus-crassus-minuta group, are discussed.

Notes: Primary trophozoites of Malpighamoeba mellificae Prell in the process of penetrating the cell membrane were found among the brush border of epithelial cells of the midgut of Apis mellifera L. They were long and slender with an average diameter of 3.68 μm. Their surfaces had some wrinkles and the cytoplasm contained some vesicles. Secondary trophozoites were found in the lumen of Malpighian tubules. Their size was variable and their shapes were highly irregular; some had pseudopodia. The smooth surface of trophozoites possessed numerous small protrusions and pits. Mature cysts were small and oval-shaped; they measured from 2.65 times 3.62-3.4 times 4.6 μm. Their surface was usually smooth but some had wrinkles.

Notes: Experiments were carried out to determine what subcellular fractions of Tetrahymena pyriformis could, after inoculation into mice, activate macrophages to kill Toxoplasma gondii in vitro. Peritoneal macrophages from mice inoculated intraperitoneally with cilia, pellicles, mitochondria, and microsomes exhibited strong toxoplasmacidal activity and had an enhanced capacity to release hydrogen peroxide (H2O2) by stimulation of a membrane-active agent as compared with resident macrophages. In contrast, macrophages from mice inoculated with macronuclei and postmicrosomal supernatant showed no toxoplasmacidal activity and a low level of H2O2 release. Similar dose response was observed on the active subcellular fractions with regard to the degree of macrophage activation. Treatment of the active subcellular fractions with heating and trypsin markedly reduced their activity.

Notes: The synthesis of DNA in two hypotrichous ciliates, Styx sp. and an amicronucleated strain of Oxytricha sp., was studied by high voltage (1000 kV) electron microscopy. High voltage EM permits use of thick sections (0.25-0.40 μm), including serial sections; thick sections produce strong autoradiographic images with relatively short exposure times. The autoradiographs show that DNA synthesis occurs in a narrow part of the rear zone of a replication band in the macronucleus. Macronuclear DNA synthesis occupies a substantial part of the interdivision interval, and micronuclear DNA synthesis in Styx sp. takes place in early prophase at a time when macronuclear DNA synthesis is in its terminal phase.

Notes: In order to answer the question of how two dissimilar flagellar motions, retraction and undulation, of the longitudinal flagellum in Ceratium tripos are regulated, the effects of cationic milieu, calcium ionophore, calcium channel blockers and some anesthetics on the motion of the longitudinal flagellum were studied. The flagellum retracted and was installed in the sulcus in high K+, high Ca2+, low Na+-ASW (artificial sea water), and low Mg2+-ASW. Although Ca2+ ionophore X537A induced the retraction, it also induced disintegration of the flagellum. The Ca2+ channel blocker, La3+, prevented the retraction effectively in high K+-ASW and in low Mg2+-ASW but did not affect it in low Na−-ASW or in high Ca2+-ASW. Ruthenium red (RR), on the other hand, prevented the retraction in high Ca2+-ASW, low Na+-ASW, and low Mg2+-ASW but did not suppress the retraction induced in high K+-ASW. The organic Ca2+ antagonist, verapamil, or the local anesthetics, dibucaine and papaveline, did not prevent the retraction effectively in any ASW. These data suggest that the flagellum retracts when external Ca2+ enters into the flagellum. The dissimilar actions of La3+ and RR suggest that there may be two different sites for Ca2+ influx which have different affinity for La3+ or RR.

Notes: The misaligned undulating membrane (mum) mutant of Tetrahymena thermophila is a non-conditional, single gene recessive mutation. The major effect of the mum mutation is the production of multiple undulating membrane (UM) fragments in the oral apparatus (OA). The ultrastructure of the UM fragments of mum OAs is identical to that of the single UM of wild-type OAs. Analysis of OA development at midbody using a combination of light microscopy of protargol-stained cells and SEM of demembranated whole cells showed that the phenotypic effect of the mum mutation first becomes evident during mid to late stage 4 and is fully manifested in early stage 5. The effect of the mutation involves a proliferation of excess basal bodies in the UM field. Subsequent events in the development of the mum OA from mid to late stage 5 are identical to those in wild-type OAs. This study suggests that the mum mutation establishes conditions that allow the production of multiple UMs and thus reveals that the UM field is competent for the complete and coordinated development of several adjacent UMs. This level of regional control is not clearly evident when a single UM is present.The comparison of development of wild-type and mum OAs required an extensive reanalysis of stages 4 and 5 of normal oral development. On the basis of current and previous observations, we propose a new and more subdivided staging system for oral development in Tetrahymena.

Notes: Sodium dodecyl sulfate (SDS) and 2-mercaptoethanol (2-ME) activated proteolytic enzymes present in extracts of Entamoeba histolytica and E. invadens; SDS (0.5%) and 2-ME (1.4 and 715 mM) doubled the enzymatic activity when assayed on a stained insoluble substrate. Urea (4 M) did not reduce this activity, suggesting that amebic proteases are stable in the above denaturant conditions. Specific reagents for sulfhydryl (-SH) groups completely inhibited proteolytic activity regardless of pH. Inhibition with alkylating agents, such as N-ethylmaleimide and iodoacetamide, was reversed with 715 mM 2-ME as was also observed with papain. We conclude from these results that the main proteolytic enzymes contained in extracts of E. histolytica and E. invadens are dependent on free thiol groups.

Notes: Paramoeba pemaquidensis Page requires attachment to a substrate in order for population growth to occur. Populations of amoebae suspended in bacterized culture media on a roller tube apparatus did not increase in size whereas amoebae maintained in stationary control tubes multiplied in number. Amoebae maintained in suspension for as long as three weeks attached and multiplied in culture when allowed to settle. This suggests that Paramoeba found suspended in the surface microlayer of the ocean may be in a “dormant” state. Implications for the survival and geographic distribution of the genus are discussed.

Notes: Zygotes of Plasmodium berghei were cultured 15–25 h in vitro to yield mature infective ookinetes. Samples taken in the first 5 h of culture were examined by electron microscopy. Meiotic figures were detected in the nuclei of the zygotes. Threadlike leptotene chromatids (chromosomes) condensed from attachment plaques on the nuclear envelope; chromatid pairing followed (zygotene), with synaptonemal complexes subsequently appearing (pachytene). These complexes persisted into metaphase but dissociated when the chromatids rapidly decondensed during anaphase. At telophase of the first meiotic division the kinetochores were retracted toward two small spindle complexes, which were found at widely separated poles in the nuclear envelope. The observations are consistent with a haploid genome of 8–10 chromosomes.

Notes: Crithidia acanthocephali and C. harmosa share the same minimal nutritional requirements as the standard C. fasciculata. Crithidia acanthocephali is exceptional, however, in that it utilizes D-ribose, free or as nucleoside (e.g. adenosine), as carbon source.

Notes: The relationship between the locomotive velocity of amoeba which had not been fed for 24 h and the concentration of the test solutions was examined. With solutions of L-amino acids, protein substances, and alcian blue 8GS, an increase in velocity began at very low concentrations, reaching a maximum at a higher concentration and as the concentration increased further, the velocity fell to zero. In contrast, no increase was observed with D-glutamic acid and β-alanine. Moreover, the velocity of well fed amoebae did not increase significantly even in L-amino acid solution. These results may suggest a correlation between orthokinesis and amoeboid phagocytosis.

Notes: Based upon light and electron microscopical observations, the feeding behavior of the ciliate Pseudomicrothorax dubius, when fed the cyanobacterium Oscilatoria formosa, is resolved into two principal phases, contact swimming and phagocytosis, the latter being separable into two steps, attachment aad ingestion. Following collision with an O. formosa filament. cells swim along the filament with their ventral cilia in contact with it during the contact swimming phase. Phagocytosis commences with the attachment of the cytostome to the filament, which initiates lysosomal streaming in the cytostomal-cytopharyngeal region. The filament then enters the cytopharynx concomitant with food vacuole formation during the ingestion step. Treatment of cells with trypsin or modification of the extracellular ionic medium inhibits the attachment step of phagocytosis but does not affect contact swimming. Behavior of cells when fed different cyanobacterial species as well as artificial food substrates is also examined. Contact swimming is a form of contact guidance since the shape of the food substrate determines the direction of cell movement. Additionally, a chemical factor may be present in or on the cyanobacteria and play a role in contact swimming. Evidence is presented that suggests that during the attachment step, two phenomena are involved: direct adhesion between cell surfaces and adhesion due to material liberated by exocytosis.

Notes: Various cations have been examined for their effects on phagocytosis. Media with high [Ca2+] and low [K+] favor phagocytosis, which is inhibited in media with high [K+], [Rb+], or [Ba2+] and low [Ca2+]. Microscopical observations of inhibited cells demonstrate that swimming behavior is not modified but they cannot perform the initial step of phagocytosis, attachment to food; when Ca2+ is added, cells attach to and ingest food, demonstrating rapid reversal of inhibition. Attachment is shown to be a linear function of the ratio [K+]/[Ca2+]1/2 or [Rb+]/[Ca2+]1/2 in the medium. The Ca2+ influx inhibitor Verapamil blocks attachment, as does La3+; the latter is believed to compete with Ca2+ for access to the Ca2+ channel. Likewise, treatment of cells with media containing no added Ca2+ inhibits attachment, and addition of 10 μM Ca2+ allows 90% of these cells to attach to and ingest food. The ionophore A23187, known to transport Ca2+ into a wide variety of cells, provokes lysosomal streaming movements typical of attachment. Based upon these observations, Ca2+ influx plays an essential role in attachment; K+ efflux also appears to be necessary since tetraethylammonium chloride blocks attachment. Treatment of cells with Tetrodotoxin, an inhibitor of Na+ transport, or suspending them in media containing no added Na+ does not affect attachment or ingestion, indicating that Na+ is not implicated in these processes. An hypothesis is presented which implicates Ca2+ in both direct adhesion and exocytosis phenomena during attachment.

Notes: An enzymatic activity that hydrolyzes O,O-diisopropylphosphofluoridate (DFP) and O-1,2,2–trimethylpropylmethyl-phosphonofluoridate (Soman) was discovered in the ciliate protozoan Tetrahymena thermophila. The enzymatic activity classifies the protein as Mazur-type similar to that found in hog kidney and Escherichia coli. The rate of hydrolysis of Soman by the Tetrahymena-extract is the highest, on a per gram of extract basis, of any eucaryote. The molecular weight is approximately 75,400 as determined by Sephacryl column chromatography. A maximum fifteen-fold purification has been achieved.Potential exists for the detoxification and one-step detection of common organofluorophosphate pollutants. Additionally, Tetrahymena should prove an easier subject for manipulation than mammalian or squid sources. Protozoa may be a potentially important source of detoxification and degradation enzymes for other environmental contaminants.

Notes: Samples of rumen contents were obtained from 13 musk-oxen living on Banks Island, N.W.T., Canada. Total protozoan numbers ranged from 38.5 to 124.6 times 104 per ml of rumen contents with a mean generic distribution of 52.4%Entodinium, 45.9%Diplodinium, and 1.7%Epidinium. A total of 18 protozoan species were found, six of which have not previously been observed in this host, i.e. D. (Diplodinium) costatum, D. (Ostracodinium) gracile, D. (Ostracodinium) trivesiculatum. D. (Eudiplodinium) medium, Epidinium quadricaudatum, and Epidinium parvicaudatum. Two new species of protozoa are described, Entodinium ovibos n. sp. and Diplodinium (Eudiplodinium) banksi n. sp.

Notes: The genus Pompholyxophrys includes amoebae which have a spherical body, fine radiating pseudopodia, and a layer of adhering siliceous “perles.” These organisms are normally regarded as a type of heliozoon. Ultrastructural examination of P. punicea reveals that those characters associated with well characterized heliozoa, such as microtubular axonemes and extrusomes, are lacking. The species has much in common with nucleariid filose amoebae with which it, and the related genus Pinaciophora, are regarded as having affinities. The species P. punicea is rare, and this study was made possible by the application of techniques developed for the ultrastructural examination of single cells. The assessment of protistan diversity and interrelationships relies heavily on the use of ultrastructural characters. Although techniques that are based on the examination of a small number of individual cells have limitations, they do allow rare organisms to be included in the re-evaluation of protistan systematics.

Notes: Proliferative kidney disease (PKD), caused by an unclassified protozoan (PKX), is reported from Pacific salmon, Oncorhynchus tshawytscha (Walbaum) and O. kisutch (Walbaum), and steelhead trout, Salmo gairdneri Richardson, held at the Mad River Hatchery in California, USA. The cumulative mortality attributed to the disease was 95, 13, and 18% respectively. The mortalities were greatest at mean water temperatures of 12-14°C during July 1983. The ultrastructure of the PKX organism and its associated pathology during clinical disease in all three species were consistent with those of the parasite in rainbow trout (Salmo gairdneri) as described in European outbreaks. Significant mortalities did not occur after August, at which time the parasite could no longer be detected in the salmon species. The steelhead continued to exhibit parasites in the kidney interstitium and epithelium and lumens of the tubules. Myxosporidan trophozoites and developing spores were also observed in the lumens of the kidney tubules of these fish. Although a mixed infection with another parasite may have occurred, evidence suggests that the myxosporidans are later stages of PKX. They were only observed in fish exposed to water with the infective stage and were particularly prominent in recovering fish. The PKX organism is similar to UBO, an unclassified protozoan of carp suspected to be an early stage of Sphaerospora renicola Dyková & Lom. Both parasites infect the blood and kidney, divide by endogeny, and are released by disintegration of the primary mother cell. The intraluminal myxosporean forms show similarities to Sphaerospora spp. in that they are monosporous and sporoblasts are formed within pseudoplasmodia. It is possible that PKX migrates to the lumen of the kidney tubule and subsequently sporulates. If the myxosporean forms are later stages of PKX, then it would belong to the phlyum Myxozoa.

Notes: Glugea stephani-infecled submucosal cells of laboratory-reared winter flounder, Pseudopleuronectes americanus, change to unique giant cell xenomas. These cells hypertrophy while the intracellular Glugea propagate to high numbers in the cytoplasm and eventually overrun the host cell. The xenomas slowly reach 20-25 muml;m (at 17°C) by day 10 after parasite invasion. These xenomas (eight often examined by electron microscopy) are coated with a mucus-like envelope onto which is attached a layer of endothelial ceils. The 10-day xenomas display (a) host cell endonuclear polyploidization and amitosis, (b) limited parasite growth and reproduction, and (c) a host cell cytoplasm structure similar to that seen in undifferentiated phagocytes. Glugea parasites do not induce obvious cell degenerative effects in 10-day xenomas; the 20-day xenomas are hypertrophied to 70-100 m̈m. These cells are characterized by (a) a host cell component denuded of endoplasmic reticulum and phagosome membrane, (b) a reduction in host cell ribosomes and the disappearance of host cell cytoskeletal assemblages, including microtubules, and (c) a significant increase in vegetative Glugea within xenomas. Evidence indicates cytoplasmic calcium of the host cell xenoma is perturbed by the intracellular Glugea; the alterations in the host cell calcium domains may be a big factor in the induction of the block of mitosis in the host cell and in the disruption in its cytoskeletal controls.

Notes: Differences in the morphology of Stylonychia vorax Stokes, 1885 and S. pustulata (Müller, 1786) Ehrenberg, 1838 recognizable in vivo are the shape, the ventral cirral pattern, the caudal cirri, and the mode of moving. The dorsal-bristle complexes are distinguishable by the length of dorsal kinety four and the spaces among the pairs of basal bodies. When the ranges of variation of different populations and clones are compared by biometric analyses, S. vorax shows a relatively stable cortical pattern whereas in S. pustulata the cortical elements are regulated depending on the size of the body and the number of adoral membranelles. In S. vorax morphogenesis begins with a proliferation of basal bodies close to the transverse cirri. In contrast, in S. pustulata, the oral primordium appears de novo between the left marginal row and the postoral cirri. All other morphogenetic events are the same for both species. In proters and opisthes the six anlagen of the frontal-ventral-transverse cirri are of different origin and evolve independently. Three anlagen of the opisthe separate from the oral primordium, two originate from the right, and one from the left postoral cirrus. Three anlagen of the proter evolve from the posteriormost cirrus in the frontal area, one from the parental undulating membranes, one from the buccal cirrus, and one from the cirrus below the buccal cirrus. The anlagen one to six generate one, three, three, three, four, and four cirri. The characteristic arrangement of the undulating membranes and the participation of only two postoral cirri in the formation of primordia provide features that distinguish between the often confused genera Oxytricha and Stylonychia.

Notes: Transmission electron microscopy and scanning electron microscopy were used to investigate the fine structure of Hepatozoon mocassini gamonts and modifications of the infected erythrocyte plasmalemma. Intraerythrocytic gamonts were contained within a parasitophorous vacuole. An electron-lucid space observed between the gamont pellicle and the membrane of the vacuole corresponded to the unstained space described in light microscopy studies. Gamonts possessed a conoid, polar ring, subpellicular microtubules, four pairs of rhoptries, micronemes, ovoid granules, mitochondria with tubular cristae, and a pellicle composed of three individual unit membranes. The conoid had an anterior diameter of 320 nm, a posterior diameter of 360 nm, and a length of 150 nm. In contrast to a report on Hepatozoon aegypti, no micropore or “canopy-like structure” was observed. The plasmalemma of infected erythrocytes exhibited two types of modifications: gross membrane deformations and knobs with an electron-dense central mass. These knobs are structurally distinct from previously described membrane excrescences.

Notes: Glugea hertwigi spores were activated to discharge sporoplasms in Medium 199 with 3% gelatin at pH 9.0; the liberated sporoplasms were transferred to a maintenance medium with 6% gelatin (pH 7.0) supplemented with 2 mM ATP and 10% (v/v) fetal calf scrum. The spherical sporoplasms (measuring 3.5-4 m̈m in diameter) had single nuclei and had a cytoplasm rich in free ribosomes. Each G. hertwigi sporoplasm was initially bounded by an external (0.1-0.2 m̈m) satellite body adjoining the plasma membrane. The satellites displayed ordered membrane and appeared to merge with the sporoplasm 15-30 min after spore discharge. The external location of the satellite (in reference to the discharged sporoplasm) seems to be part of the normal sequence of events under the in vitro conditions provided. The surface of G. hertwigi sporoplasms does not bear an obvious surface coat; however, our cytochemical observations indicate the plasma membrane of the sporoplasm was somewhat responsive to concanavalin A-peroxidase, colloidal iron, and native ferritin. During the short term incubations of sporoplasms with ferritin, the particles permeated membrane channels extending into the sporoplasm cytoplasm.

Notes: The sequential development of Ichthyophthirius multifiliis Fouquet, 1876 at 22°C was monitored from detachment from the fish through to infective tomites using one-dimensional polyacrylamide gel electrophoresis. Methods are described for synchronizing I. multifiliis cells to permit the collection of large numbers of cells at stages during mitotic division. The electrophoretic protein patterns of the encysted stage differ from those of the tomite stage as well as from those of the other measured intervals.

Notes: Electrophoretic separation of the water-soluble proteins from seven Acanthamoeba strains, followed by lactic dehydrogenase (LDH) visualization by the zymogram method, has indicated a single LDH activity in each extract. To account for this observation, three alternatives are considered: inadequate electrophoretic resolution, two gene-encodement of LDH with constraints relating to subunit assembly or oligomer stability, and one gene-LDH encodement.

Notes: We explored the requirements of inorganic phosphate (Pi), the incorporation of 32P-orthophosphate (32Pi), and the occurrence of inorganic polyphosphate (polyP) in axenic Entamoeba cultures. Maximal population densities and growth rates of Entamoeba histolytica trophozoites were attained in complete TP-S-1 medium. As 32Pi concentration was increased in the medium, its own incorporation and the culture growth rate were progressively inhibited, especially in Pi-deficient medium. PolyP grains were found in the cytoplasm and occasionally in the nuclear membrane of E. histolytica, E. histolytica-like, E. invadens, and E. moshkovskii trophozoites.

Notes: Spore polypeptide profiles of Nosema locustae and an unidentified microsporidium infecting Aulocara elliotti and Psoloessa delicatula are compared with sodium dodecyl sulfate (SDS) polyacrylamide gel electrophoresis (PAGE). Unique spore polypeptide profiles are not detected for the unknown microsporidium from A. elliotti and P. delicatula whereas these profiles are distinctly different from N. locustae spore polypeptide profiles. These results indicate that the microsporidium is not a polymorphic form of N. locustae but a separate species.

Notes: Membrane fragments from trypomastigote forms of Trypanosoma cruzi inhibited the association of intact trypomas- tigotes with rat heart myoblasts whereas a similar preparation from non-invasive epimastigotes did not. Furthermore, killed trypo- mastigotes bound to the host cell surface and prevented the attachment of living organisms. Conversely, the extent of association of killed parasites with the host cells was reduced by the presence of living flagellates. These results suggest the presence of a distinct structure(s) on the surface of rat heart myoblasts to which infective forms of T. cruzi can bind.

Notes: On the basis of a review of the approximately 4300 species of apicomplexan protozoa, the following new species, new names, new combinations, and emendations are given: NEW GENUS,Erhardovina; NEW SPECIES,Ascogregarina polynesiensis, Eimeria golemanskii, Isospora tamariscini; NEW NAME,Gregarina kazumii; NEW COMBINATIONS,Ascogregarina brachyceri (Purrini, 1980),Erhardovina euzeti (Lipa, 1981),E. scutovertexi (Erhardová, 1955),Haemorhormidium batrachi (Chaudhuri & Choudhury, 1983); EMENDATIONS,Selenidium francianum(Arvy, 1952) Tuzet & Ormières, 1965,Pyxinioides bolitoides D. P. Henry, 1938,P. japonicus H. Hoshide, 1951,P. kamenote H. Hoshide, 1951,P. kurofuji H. Hoshide, 1951,P. oshoroensis H. Hoshide, 1951,P. pugetensis D. P. Henry, 1938, Gregarina levinei Haldar & Sarkar, 1980,Retractocephalus halticae Haldar, Chakraborty & Kundu, 1982,Cnemidospora schizophylli Tuzet & Guerin, 1947,Grebneckiella indica (Merton, 1911) Watson, 1916,Quadruspinospora atractomorphae Haldar & Chakraborty, 1978,Haemogregarina acipenseri Nawrotzky, 1914,H. lobianci Yakimov & Kohl-Yakimov, 1912,H. yakimovikohlae Wladimiroff, 1910,Hepatozoon luehi (Sambon, 1909) Pessoa, Cavalheiro & de Souza, 1970,Eimeria beyerae Ovezmukhammedov, 1977, E. (?) gigantea (Labbé, 1896) Reichenow, 1921, E. (?) labbei Hardcastle, 1943, E. rufi Prasad, 1960, E. (?) scylii (Drago, 1902) Levine & Becker, 1933, Isospora corvi Ray, Shivnani, Oommen & Bhaskaran, 1952,I. melopsittaci Bhatia, Chauhan, Arora & Agrawal, 1973, I. seicerci Ray, Shivnani, Oommen & Bhaskaran, 1952,I. stomatici Chakravarty & Kar, 1944,I. triffitae Nukerbaeva & Svanbaev, 1973,Wenyonella mackinnonae Misra, 1947,Octosporetla sanguinolentae Ovezmukhammedov, 1975,Lankesterella millani Alvarez Calvo, 1975,Sarcocystis woodhousei Dogel', 1916,Haemoproteus lari Yakunin, 1972, Babesia ninakohlyakimovae (Yakimoff & Shokhor, 1916),Theileria ninakohlyakimovae (Yakimov, 1916) Krylov, 1974,Haemohormidium batrachi(Chaudhuri & Choudhury, 1983).

Notes: Publications on the coccidia of certain invertebrates are reviewed and two new taxonomic-nomenclatural combinations are introduced: Alveocystis macrocoronata (Lüling, 1942) n. comb., in hosts Priapulus caudatus and Halicryptus spinulosus (Priapuloidea); and A. gugleri (Wacha, 1981) n. comb., in Triodopsis albolabris (Mollusca).

Notes: Macronuclear DNA synthesis normally continues until late in the cell cycle in Paramecium; however, blockage of macronuclear DNA synthesis after 0.72 in the cell cycle does not alter the occurrence or timing of the subsequent cell division. When DNA synthesis is blocked after cells have reached the transition point, macronuclear DNA content at the following division is reduced to about 75% of the normal level. The point at which macronuclear DNA synthesis is no longer required for division corresponds to the beginning of micronuclear mitosis and the early stages of oral morphogenesis.

Notes: By serial sectioning and 3D reconstruction we have been able to demonstrate that the type of system for hemoglobin digestion in two strains of Plasmodium berghei, N and RC, is dependent on the maturity of the host cell. In parasites growing in erythrocytes, both systems for the endocytosis of hemoglobin—micropinocytosis and the cytostomal system (i.e. a cytostome budding a cytostomal tube that releases food vacuoles)—are fully functional and produce a great quantity of residual pigment. Parasites growing in reticulocytes have a disrupted cytostomal system; no tube is formed and only food vacuoles are visible in their cytoplasm. Residual pigment is smaller in size and in quantity. The reduced quantity of pigment in reticulocytes is explained by our observation of the exocytosis of pigment. We propose a hypothesis that relates the process of degradation of hemoglobin to the maturity of the host cell and a possible mechanism of protection against chloroquine, a drug known for its affinity for malarial pigment.

Notes: The strain of ameba, culture incubation temperature, and phase of ameba growth affected the number of amebostomes present on amebae of Naegleria fowleri. Serial passage of N. fowleri through mice decreased the average number of amebostomes. Amebostomes were shown to be functional by their ability to engulf yeast cells.

Notes: Time-lapse phase-contrast videomicroscopy revealed that the psudopodial network of two allogromiid foraminiferense display an invasive behaviour, previously undescribed, which I term Skyllocytosis (Greek: skylo—to rend, tear, pluck). When these networks encounter an interface between a gelatin/agar overlay and a glass substratum, portions of the overlay are penetrated and partly surrounded by reticulopodia. By the coordinated activity and contraction of these reticulopodia, small segments of the overlay are ripped away. Manageable portions of the overlay are subsequently transported towords the cell body. In carnivorous foraminifera skyllocytosis may account for the removal of soft, autolysed tissues from dead invertebrate prey.

Notes: The digestive-lysosomai system in Telrahymena has been extensively studied; however, the various vacuole stages and the existence of a required processing period prior to defecation have not been clearly denned, in this study the presence of such a required processing period and the rate of DV defecation in Tetrahymena thermophila were determined. Like the cycle in Paramecium. a digestive cycle in Tetrahymena consisted of two periods: the processing period was 45 min and the defecation period was ˜2 h, making the complete cycle ˜3 h. During the defecation period vacuole egestion followed the kinetics of a first-order rate reaction and had a rate constant of 0.0187/min and a t1/2 of 37 min (82 min into the cycle). Using the naphthol AS-TR phosphate-hexazotized rosanilin method to visualize acid phosphatase activity at the light microscopic level, DVs became positive beginning at 10 min. The number of positive DVs increased to a maximum of 13% when DVs were 20-min old and declined to 5-7% beyond 30 min. Although dichloroisoproterenol (DCI) has been reported by others to stimulate vacuole defecation, we found it inhibited the defecation rate. The extent of inhibition depended on the age of the DVs when exposed to DCI. Vacuole formation was completely blocked in cells pre-exposed to 40 μ DCI for only 10 min; however, upon further exposure, cells could recover from this inhibition. The time required for complete recovery increased with increasing DCI concentrations. If DCI was given to cells simultaneously with latex beads, it was found to exert a dose-dependent inhibitory effect on DV formation. These results showed the heterophagic pathway of the digestive-lysosomal system in Telrahymena to be similar to that of Paramecium, though it was less efficient in the former cell.

Notes: Crithidia fasciculata and Phytomonas davidi catabolize tryptophan (TRP) to indole-3-ethanol, which was identified by both thin layer and gas chromatography. The catabolic pathway involved in this metabolic conversion is suggested to be similar to that proposed for other members of the family Trypanosomatidae. Although this catabolism occurs at both 25° and 37°C, the catabolic rate is greater at 37°C, a non-permissive growth temperature. Conditions that inhibit protein synthesis would appear to favor the catabolism of tryptophan to indole-3-ethanol. The possible importance of this catabolic pathway to these organisms is discussed.

Notes: The ribosomal RNA from several stocks of the genera Leishmania and Trypanosoma were studied by gel electrophoresis, sedimentation on sucrose density gradients and RNA/DNA hybridization experiments. Three major components were observed after electrophoresis in polyacrylamide gels (PAGE-SDS), the relative molecular masses being respectively: X1= 0.83 megadaltons, X2= 0.63 megadaltons and X3= 0.54 megadaltons for Leishmania RNA; and X1= 0.86 megaldaltons, X2= 0.78 megadaltons, and X3= 0.58 megadaltons for Trypanosoma RNA. Depending upon the isolation procedure, a fourth component. X0= 1.2 megadaltons (26S), became evident. The later component was purified from Leishmania brasiliensis (Y) by centrifugation on a linear 15-30% sucrose density gradient. This component, after heat denaturation and PAGE-SDS, gave rise to two bands coinciding in molecular mass with those of X2 and X3 indicating that these components are part of the large ribosomal subunit whereas X1 belongs to the small one. The above mentioned differences in mobilities of components X1 and X2 between the two genera were no longer observed after electrophoresis in denaturing agarose-formaldehyde gels, suggesting secondary structural differences among these RNA species. Hybridization experiments with L. brasiliensis (Y) DNA showed that both RNA types compete equally well for the ribosomal sites in this DNA, and that L. brasiliensis (Y) rRNA recognizes the ribosomal sites in DNA of Trypanosoma cruzi (EP), thus indicating that no gross changes occurred in their nucleotide sequences during evolution.

Notes: A rapid and simple method for the purification of amastigotes of Trypanosoma cruzi from spleens of infected mice is described. A protein A-Scpharose 4B immunoadsorbent column bound with antisera to epimastigotes of T. cruzi was used to purify the tissue forms of this parasite. Host cells and debris are not retained, and parasites can be eluted in high yields and purity. Studies of surface glycoproteins and glycolipids of the purified amastigotes with 18 lectins of various specificities revealed the presence on the parasites of receptors for N-acetylglucosamine, N-acetylgalactosamine, D-galactose, and D-mannose binding lectins.

Notes: Erythrocytes infected with Plasmodium falciparum bind specifically to cultured endothelial cells and to a line of amelanotic melanoma cells. We have fixed endothelial cells and amelanotic melanoma cells in various ways and determined whether the fixed cells were still able to bind infected erythrocytes. Only cells fixed with 1.0-2.5% formalin in phosphate-buffered saline continued to bind infected erythrocytes as well as unfixed cells. The mechanism of binding to fixed and unfixed cells appeared to be identical for the following reasons. First, erythrocytes infected by parasite strains that bound to unfixed cells also bound to fixed cells while those that did not bind to unfixed cells did not bind to fixed cells. Second, immune serum that inhibited binding to unfixed cells also inhibited binding to fixed cells. Third, electron microscopy showed that knobs were the points of attachment between infected erythrocytes and both fixed and unfixed melanoma cells. Fixed cells gave reproducible results over at least 2 months. Thus, we have developed a simplified, reproducible assay for measuring binding of P. falciparum-infected erythrocytes to target cells.

Notes: Crithidia fasciculata was used as a model trypanosomatid to study the possible existence of genetic recombination in this group of protozoa. The approach was based on the ability to select a variety of mutants on agar plates. Following mutagenesis of wild type cells by nitrosoguanidine or ethylmethanesulfonate, stable mutant phenotypes were obtained. These included mutants resistant to the drugs actinomycin D, 6-azauracil, 6-azauridine, and 5-fluorouracil, auxotrophs and colony morphology mutants. Following mixed growth of pairs of drug-resistant mutants on selective media, isolates exhibiting stable recombinant phenotypes were obtained. The data presented suggest that 1) Crithidia undergoes some type of genetic recombination and 2) Crithidia must be diploid at some time during this process.

Notes: Exoantigens of Trypanosoma cruzi were produced in experimentally infected BALB/c mice. The exoantigens were detected by the counterimmunoelectrophoresis method (CIE), with antisera raised in rabbits by immunization with total homogenates of culture forms of ***T. cruzi in plasma from ***field animals obtained by centrifugation and filtration. Control experiments indicated that exoantigens are not somatic components of T. cruzi leaked during the preparative procedure. Exoantigens were detected in male and female mice, 11-90 days old, between 6 and 60 days of infection, and in all mice with patent parasitemia. After 13 days of infection, mice developed antibodies to exoantigens; by CIE up to three populations of antibodies were revealed in different groups of animals. In mice between 13 and 60 days of infection, the coexistence of exoantigens and homologous antibodies was also observed. The exoantigens are not strain specific since a cross reactivity between antigens from three strains of T. cruzi (Tulahuén, Higueras, and Alejandro) was seen. Finally, the presence of antibodies to exoantigens in humans with chronic Chagas’ disease was demonstrated.

Notes: The interaction of Eimeria falciformis sporozoites with the intestinal epithelium and with the intestinal contents from the cecum and colon of normal and specifically immunized mice was studied by light (LM) and scanning electron (SEM) microscopy. Fecal (FM) and enterocyte-associated (EAM) mucus were removed from the cecum and colon of normal mice and mice that had been immunized 1, 6, 12, or 20 days earlier with a series of oral inoculations of E. falciformis oocysts. Sporozoite-specific IgA, but neither IgM nor IgG, was detected by the immunofluorescent antibody test in FM and EAM from immunized mice. No sporozoite-specific immunoglobulin was detected in normal mice. When examined by LM, sporozoites exposed to all FM and EAM preparations exhibited greater motility and excystation from sporocysts. At 4 h after incubation in FM or EAM from normal or immune mice, about 10% of the sporozoites appeared damaged, being non-motile and non-refractile. Immune FM and EAM caused agglutination of sporozoites and sporocysts and oocyst walls of E. falciformis. but did not agglutinate those of E. ferrisi. Scanning electron microscopy of in vitro interactions between E. falciformis sporozoites and intestinal contents revealed that sporozoites exposed to immune EAM were coated with particulate material whereas those exposed to normal EAM were relatively clean. Sporozoites exposed to immune FM were usually embedded within the mucus whereas those exposed to normal FM were situated on top of the mucus. No significant differences occurred between the length/width (L/W) ratios of sporozoites incubated in normal FM and EAM or in PBS. Sporozoites exposed to immune FM had significantly greater L/W ratios than those exposed to normal FM whereas those exposed to immune EAM had significantly shorter L/W ratios than ones exposed to normal EAM. Few of the sporozoites observed on the luminal surface of the colon and cecum of normal mice were covered by mucus and none was altered in shape or showed pellicular damage. Only a few sporozoites were observed on the luminal surface of the colon and cecum of immunized mice. Most of these were covered by mucus and some exhibited pellicular alterations.

Notes: The cytopathogenicity of Naegleria fowleri strain LEE (ATCC-30894) for cultured rat neuroblastoma cells (B-103) has been investigated. Both live N. fowleri amoebae and Naegleria lysates added to 51Cr-labeled B-103 cells caused release of radiolabel, which was dependent upon the ratio of amoebae to target cells or to the lysate concentration. Lysates of N. fowleri strains LEE, NF-66, NF-69, and HB-4 were equally injurious to B-103 target cells whereas lysates of strains 6088 and KUL were less cytotoxic. Highly pathogenic mouse-passaged strain LEE were less cytotoxic than axenically grown amoebae. Maximum cytotoxicity was observed in lysates from amoebae in late exponential or early stationary phase of growth. Cytopathogenicity of lysates was reduced after heating at 44°C for 60 min or at 60°C for 30 min. Cytotoxicity was stable during storage at 4°C or at −20°C for 26 h. Neither live amoebae nor lysates injured B-103 target cells at 4°C. Live amoebae and lysates injured B-103 by a time, temperature, and concentration dependent process.

Notes: Paramoeba invadens n. sp. is described from sea urchin tissues and from culture. Amoebae are 20–40 μm in length with an irregular hyaline region producing short subpseudopodia. One parasome per cell is present. The parasome is bipolar with basophilic, Feulgen-positive poles and a Feulgen-negative median segment. The fine structure of the parasome resembles those in other species of Paramoeba and the surface of the amoeba is plain with no hairs or scales. Amoebae dislodged from tissues often adopt a semifloating form; floating forms have been seen in culture. Amoebae circulate within lumina of the water vascular system and can migrate readily into or out of tissues.

Notes: In the examination of 90 Chondrostoma polylepis caught in the Esla River, a coccidium of the genus Goussia was found in the swimbladder, peritoneum, kidney, and ureter. It is described as Goussia polylepidis n. sp. and its taxonomic affinities are discussed. Data on its prevalence and seasonality are also given.

Notes: Eight isolates of Naegleria australiensis were obtained from a small lake in Tulsa, Oklahoma. The eight strains were isolated during the hot summer months of July through September, when water temperatures ranged from 27 to 33°C. All eight isolates were pathogenic for mice. The mean time to death for mice was 10 days (range 6–13 days). This pathogenic free-living ameba has not before been reported from the United States or the Western Hemisphere.

Notes: Isospora dawadimiensis n. sp. is described from the jerboa, Jaculus jaculus, from Dawadimi, Saudi Arabia. Sporulated oocysts of I. dawadimiensis n. sp. were ovoidal or nearly subspherical 22–26.5 times 20.5–22 μm (24.4 times 21.4 μm). Oocyst wall had one layer. Micropyle, oocyst residuum, and polar granule were absent. Sporocysts were ellipsoid 12–16.5 times 9–10.5 μm (14.6 times 9.9 μm). Sporocyst residuum was present. The sporocysts lack a Stieda body. Sporozoites 8–11 times 2–3 μm (10 times 2.6 μm) were sausage-shaped, slightly curved, and tapered at one end.

Notes: Hausmann, K. (with the collaboration of M. Mulish and D. J. Patterson) 1985. Protozoologie.

Notes: A new species of microsporidium (phylum Microspora), Microsporidium novacastriensis n. sp., from the grey field slug, Deroceras reticulatum, is described on the basis of light and electron microscope studies. Meronts are spherical at first, then become irregular as nuclear number increases. Sporonts are tubular or ribbon-like and divide unevenly to produce sporoblasts and then spores of varying lengths. Sporogonial stages are enclosed in a vesicle by a subpersistent membrane of uncertain origin. Fresh spores measure 3.5 by 2.08 μm and are produced in clusters of 12 to 120. The parasite infects only the intestinal epithelium of the slug. The new species is compared to microsporidia of other gastropod molluscs and to other microsporidia of similar developmental pattern and morphology.

Notes: During the spring of 1984, the ciliate Balantidium prionurium n. sp. was collected from the intestinal lumen of the herbivorous surgeonfish, Prionurus punctatus, from the Gulf of California. The symbionts were found in five fish from two well separated collection sites. Morphostatic specimens average 51 μm x 42 μm, and thus fall within the size range of several other fish balantidia. But the presence of Balantidium in a saltwater fish has not been reported, and such a host difference alone supports at least provisional recognition of this organism as a new species.

Notes: Hausmann, K. & Patterson, D. J. 1983. Taschenatlas der Einzeller: Protisten. Arten und Mikroskopische Anatomie.