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Morphological and molecular characteristics of a species of Hepatozoon Miller, 1908 (Apicomplexa: Adeleorina) from the blood of Isoodon obesulus (Marsupialia: Peramelidae) in Western Australia.

Examination of blood films as part of a study to assess the health status of the southern brown bandicoot Isoodon obesulus (Shaw) in Western Australia revealed the gamonts of a haemogregarine parasite in some samples, the first to be recognised in a bandicoot in this state. Light microscope morphological characteristics and partial sequence of the 18S rRNA gene were used to describe these organisms. Morphological characters did not differentiate the organism in the current study from previously reported Hepatozoon peramelis (Welsh & Dalyell, 1909). Phylogenetic analysis has not previously been reported for any species of Hepatozoon from Australian marsupials and consequently could not be used to confirm the identity of the organism in the current study as that described in the 1900s. If this organism is H. peramelis, then it has a wide distribution, being found in three species of bandicoot, in western and eastern Australia and the in island state of Tasmania.

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Phylogenetic position of the genus Perkinsus (Protista, Apicomplexa) based on small subunit ribosomal RNA.

Parasites of the genus Perkinsus destroy marine molluscs worldwide. Their phylogenetic position within the kingdom Protista is controversial. Nucleotide sequence data (1792 bp) from the small subunit rRNA gene of Perkinsus sp. from Anadara trapezia (Mollusca: Bivalvia) from Moreton Bay, Queensland, was used to examine the phylogenetic affinities of this enigmatic genus. These data were aligned with nucleotide sequences from 6 apicomplexans, 3 ciliates, 3 flagellates, a dinoflagellate, 3 fungi, maize and human. Phylogenetic trees were constructed after analysis with maximum parsimony and distance matrix methods. Our analyses indicate that Perkinsus is phylogenetically closer to dinoflagellates and to coccidean and piroplasm apicomplexans than to fungi or flagellates.

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Acute osmotic tolerance of cultured cells of the oyster pathogen Perkinsus marinus (Apicomplexa:Perkinsida).

Cultured Perkinsus marinus cells were exposed for 24 hr to salinities of 0, 3, 6, 9, 12 and 22 ppt at temperatures of 1, 5, 10, 15 and 28 degrees C in artificial seawater (ASW) and to the same salinities at 28 degrees C in ASW with the osmotic concentration adjusted with sucrose to the equivalent of 22 ppt. At 28 degrees C mortality increased as salinity decreased below 22 ppt. Mortality was greater than 99% at 0 ppt and greater than 90% at 3 ppt. Mortality was 70% at 6 ppt, 43% at 9 ppt and 20% at 12 ppt. Mortality was low (< 5%) and equal to that at 22 ppt in all treatments where osmotic concentration was maintained with sucrose. Mortality occurred rapidly, within 5 min of exposure to experimental conditions. In the region where mortality was most sensitive to salinity changes (6-12 ppt), lower temperature caused an increase in mortality, but the temperature effect was significant only at 9 ppt.

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The ultrastructure and reproduction of Amphiamblys capitellides (Microspora, Metchnikovellidae), a parasite of the gregarine Ancora sagittata (Apicomplexa, Lecudinidae), with redescription of the species and comments on the taxonomy.

The ultrastructural cytology and reproduction of the hyperparasitic microsporidium Amphiamblys capitellides (Caullery and Mesnil, 1897) is described. Merogonial reproduction was not observed. The sporogony comprises two sequences: a sac-bound sporogony in close contact with the cytoplasm of the host and a free sporogony in parasitophorous vacuoles. The free sporogony, which probably precedes the sac-bound, yields a small number of rounded spores. The sac-bound sporogony is polysporoblastic, generating two rows of elongated spores. All stages have isolated nuclei. Both spore types have an extrusion apparatus of the metchnikovellidean type, with a polar sac devoid of anchoring disc, a polar filament with one manubroid and one bulbous part, and a posterior semicircular membrane fold enclosing rounded or tubular structures. Hosts are gregarines of the species Ancora sagittata living in the intestine of polychaetes of the genus Capitella, probably the species Capitella giardi. The cytology, life cycle and classification are discussed. The species is redescribed and the diagnosis of the genus Amphiamblys Caullery and Mesnil, 1914 is emended.

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Effect of Diplocystis tipulae Sherlock (Eugregarinida: Apicomplexa), a coelomic gregarine pathogen of tipulids, on the larval size of Tipula paludosa Meigen (Tipulidae: Diptera).

This study demonstrates the debilitative effect of a coelomic gregarine, Diplocystis tipulae, on Tipula paludosa. The larvae were provided contaminated fresh grass leaves from a field where 40.0% of T. paludosa larvae were infected by this pathogen. Resultant infected larvae were separated into four groups according to infection level. Analysing their weights, lengths, and weight/length ratios showed that larval size decreased as infection level increased. Differences, especially at the lower and upper levels of the infection levels, were statistically significant. It was concluded that infection by D. tipulae affected the size of T. paludosa larvae resulting in smaller individuals.

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Prevalence and seasonality of Ascogregarina culicis (Apicomplexa: Lecudinidae) in natural populations of Aedes aegypti (Diptera: Culicidae) from temperate Argentina.

Ascogregarina infections from South America were recently documented in Brazil and Argentina. The aim of this study was to report our recent findings on the prevalence and seasonality of Ascogregarina culicis in Aedes aegypti adults from temperate Argentina. Between December 2003 and May 2005, 391 females of Ae. aegypti were captured in two areas of Greater Buenos Aires. Overall prevalence of A. culicis was 21.2% (83/391), and a significant difference was observed between both areas (28.4% vs. 8%). Infected Ae. aegypti were found from November to May, with highest values in March (24-37%). Parasite prevalence and host abundance showed similar seasonal patterns. Our observations suggest a widespread infestation of A. culicis among Ae. aegypti populations from temperate Argentina.

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Analysis of gene expression during development: lessons from the Apicomplexa.

Apicomplexans are responsible for significant human and animal disease worldwide, including malaria and toxoplasmosis. Herein we summarize recent advances in gene expression analysis in these eukaryotic pathogens, especially with respect to their developmental biology, and discuss the impact this work may have on the development of new vaccines and chemotherapeutics.

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Phylogeny of marine Gregarines (Apicomplexa)--Pterospora, Lithocystis and Lankesteria--and the origin(s) of coelomic parasitism.

Gregarines constitute a large group of apicomplexans with diverse modes of nutrition and locomotion that are associated with different host compartments (e.g. intestinal lumena and coelomic cavities). A broad molecular phylogenetic framework for gregarines is needed to infer the early evolutionary history of apicomplexans as a whole and the evolutionary relationships between the diverse ultrastructural and behavioral characteristics found in intestinal and coelomic gregarines. To this end, we sequenced the SSU rRNA gene from (1) Lankesteria abbotti from the intestines of two Pacific appendicularians, (2) Pterospora schizosoma from the coelom of a Pacific maldanid polychaete, (3) Pterospora floridiensis from the coelom of a Gulf Atlantic maldanid polychaete and (4) Lithocystis sp. from the coelom of a Pacific heart urchin. Molecular phylogenetic analyses including the new sequences demonstrated that several environmental and misattributed sequences are derived from gregarines. The analyses also demonstrated a clade of environmental sequences that was affiliated with gregarines, but as yet none of the constituent organisms have been described at the ultrastructural level (apicomplexan clade I). Lankesteria spp. (intestinal parasites of appendicularians) grouped closely with other marine intestinal eugregarines, particularly Lecudina tuzetae, from polychaetes. The sequences from all three coelomic gregarines branched within a larger clade of intestinal eugregarines and were similarly highly divergent. A close relationship between Pterospora schizosoma (Pacific) and Pterospora floridiensis (Gulf Atlantic) was strongly supported by the data. Lithocystis sp. was more closely related to a clade of marine intestinal gregarines consisting of Lankesteria spp. and Lecudina spp. than it was to the Pterospora clade. These data suggested that coelomic parasitism evolved more than once from different marine intestinal eugregarines, although a larger taxon sample is needed to further explore this inference.

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Lipid biology of Apicomplexa: perspectives for new drug targets, particularly for Toxoplasma gondii.

Development of effective therapies for intracellular eukaryotic pathogens is a serious challenge, given the protected location of these pathogens and the similarity of their biology to that of the host. Identifying cellular processes that are unique to the parasite is therefore a crucial step towards defining appropriate drug targets. In the case of the apicomplexan parasite Toxoplasma gondii, the need to find alternative treatments is imperative because of the poor tolerability and frequent side-effects associated with existing therapeutic strategies. The discovery that the parasite uses lipid synthetic pathways which are different from, or absent in, the mammalian host is now driving a renewed interest in T. gondii lipid biology. Recent achievements in this field are promising and suggest that the elucidation of lipid pathways will provide new opportunities for designing potent antiparasitic strategies.

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The glideosome: a molecular machine powering motility and host-cell invasion by Apicomplexa.

The apicomplexans are obligate intracellular protozoan parasites that rely on gliding motility for their migration across biological barriers and for host-cell invasion and egress. This unusual form of substrate-dependent motility is powered by the "glideosome", a macromolecular complex consisting of adhesive proteins that are released apically and translocated to the posterior pole of the parasite by the action of an actomyosin system anchored in the inner membrane complex of the parasite. Recent studies have revealed new insights into the composition and biogenesis of Toxoplasma gondii myosin-A motor complex and have identified an exciting set of small molecules that can interfere with different aspects of glideosome function.

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Apical organelles and host-cell invasion by Apicomplexa.

Host-cell invasion by apicomplexan parasites involves the successive exocytosis of three different secretory organelles; namely micronemes, rhoptries and dense granules. The findings of recent studies have extended the structural homologies of each set of organelles between most members of the phylum and suggest shared functions of each set. Micronemes are apparently used for host-cell recognition, binding, and possibly motility; rhoptries for parasitophorous vacuole formation; and dense granules for remodeling the vacuole into a metabolically active compartment. In addition, gene cloning and sequencing have demonstrated conserved domains, which are likely to serve similar functions in the invasion process. This is especially true for microneme proteins containing thrombospondin-like domains, which are likely to be involved in binding to sulphated glycoconjugates. One such protein was recently shown to be required for the motility of Plasmodium sporozoites. These molecules have been shown to be shed on the parasite and/or cell surfaces during the invasion process in Plasmodium, Toxoplasma and Eimeria. For rhoptries and dense granules, the association between exocytosed proteins and the parasitophorous vacuole membrane had been analyzed extensively in Toxoplasma, as these proteins are likely to play a crucial role in metabolic interactions between the parasites and their host cells. The development of parasite transformation by gene transfection has provided powerful tools to analyze the fate and function(s) of the corresponding proteins.

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The life cycle of Gregarina ronderosi n. sp. (Apicomplexa: Gregarinidae) in the Argentine grasshopper Dichroplus elongatus (Orthoptera: Acrididae).

Gregarina ronderosi n. sp. is described based on life cycle observations conducted on nymphs and adults of its natural host, the grasshopper Dichroplus elongatus. Following ingestion of oocysts by the host, parasite development occurs between the epithelium and the food mass in the midgut and gastric caeca. Gametocysts are liberated in the faeces. Natural prevalence in the type locality, Girondo, northwestern Buenos Aires Province, was 39.7% (n=131). The earliest trophozoites seen were small (< or = 10 microm), somewhat ovoid, unsegmented bodies. Fully developed trophozoites (the body is divided into epimerite, protomerite, and deutomerite) were slender, with conical or globular epimerites in attached or unattached forms, respectively. Trophozoites varied greatly in size [total length: 10.4-275.1 microm; mean (+/-S.E.): 126.3+/-78.9]. Gamonts, which were the most common stages observed and filled the midgut and gastric caeca in grasshoppers kept in rearing rooms, had a stocky appearance and also varied greatly in size (total length: 80-348 microm; 205+/-13). Association of gamonts was precocious, biassociative, and caudofrontal. Gametocysts were spherical and highly variable in size (96-376 microm in diameter; 202.8+/-52.5), and normally have 14 sporoduct basal discs. Everted sporoducts were up to 60 microm long. Oocysts were uniformly doliform in shape, measured (5+/-0.08 by 3.2+/-0.06 microm) and contained eight sporozoites. Wall reinforcements (carinae) were present. No infection resulted in experimentally inoculated Locusta migratoria, which is a host of Gregarina acridiorum. G. ronderosi is strikingly similar to G. acridiorum, but has larger oocysts.

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Susceptibility of Aedes aegypti and Aedes albopictus larvae to Ascogregarina culicis and Ascogregarina taiwanensis (Apicomplexa: Lecudinidae) from Florida.

The susceptibility of Aedes aegypti to Ascogregarina culicis and Aedes albopictus to Ascogregarina taiwanensis was examined with mosquito and parasite strains from Tampa, FL. When each host was bioassayed with its natural gregarine, the infection intensity indicated that Ae. aegypti was 59% more susceptible to A. culicis (87 gamonts/larva) than Ae. albopictus to A. taiwanensis (47 gamonts/larva). Infections in single and mixed host populations exposed to 100 oocysts/larva of one and both parasites demonstrated that Ae. aegypti harbors higher A. culicis gamont loads than Ae. albopictus of A. taiwanensis. In dual gregarine exposures of single host populations, the A. culicis infection intensity in Ae. aegypti was reduced by approximately 50%. A. taiwanensis exhibited the same capability of infecting Ae. albopictus in single and dual exposures. In mixed host populations there were no cross infections, but A. taiwanensis in Ae. albopictus produced an infection intensity of approximately 70% lower than that of A. culicis in Ae. aegypti.

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Apical organelles of Apicomplexa: biology and isolation by subcellular fractionation.

The apical organelles are characteristic secretory vesicles of Plasmodium, Toxoplasma, Cryptosporidium and other apicomplexan organisms. They consist of rhoptries, micronemes and dense granules. Recent research has provided much new data concerning their structure, contents, functions and development. All of these organelles contain complex mixtures of proteins, with broad homologies as well as differences in molecular structure between species and genera. Many of the proteins interact with host cell membranes, and are thought to mediate selective adhesion to host cells as well as membrane modification during intracellular invasion. Micronemal proteins are important in the initial selection of host cells, and in enabling gliding motility of the parasites, while rhoptries appear to be more important in parasitophorous vacuole formation. Dense granules are involved predominantly in modifying the host cell after invasion. Research into apical organellar composition and function depends on accurate assignment of molecular identity. This requires the simultaneous application of several complementary approaches including immunolocalisation by light- and electron-microscopy, subcellular fractionation, and transgene expression. The merits and limitations of these different types of approach are discussed, and the importance of cell fractionation methods in characterising apical organelle proteins is stressed.

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The life cycle of Ascogregarina taiwanensis (Apicomplexa:Lecudinidae).

A number of features, including large size of the trophozoite, host-specificity, developmental synchrony and evident migration behavior, of Ascogregarina taiwanensis make it suitable as a model for studies of parasite-host relationships. In this article, Wei June Chen describes the life cycle of As. taiwanensis; and raises some areas for future work, including the interaction between the parasite and its host, inhibition of sexual reproduction by non-natural hosts and their mutual effects during the development.

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