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Eimeria atheridis n. sp. (Apicomplexa: Eimeriidae), a new coccidium from the western bush viper Atheris chlorechis (Pel, 1851) from tropical Africa.

Coprological examination of nine bush vipers Atheris chlorechis imported from Ghana revealed the presence of a new coccidian species belonging to Eimeria Schneider, 1875. Thin walled oöcysts of Eimeria atheridis n. sp. are spherical to slightly subspherical, 22.8 (19-26) x 22.5 (19-25) microm, without micropyle, polar granule and oöcyst residuum. Sporocysts are elongately ellipsoidal, 17.1 (15-19) x 7.5 (6-8) microm, with a dome like, relatively flat Stieda body. Sporozoites possess two refractile bodies and distinct transversal striation. Based on the presence of a Stieda body the species described herein clearly belongs to the Eimeria ( sensu stricto ).

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A survey of coccidian infection of freshwater fishes in South Africa, with the description of Goussia anopli n. sp. (Apicomplexa: Eimeriidae).

Seventy-seven specimens of seven freshwater fish species harvested in four sites from rivers and ponds of the Gauteng, North West and Limpopo Provinces (South Africa) were surveyed for coccidian infections. Two fish species were infected with apicomplexans belonging to Goussia Labbé, 1896. In banded tilapia Tilapia sparrmanii Smith unsporulated oöcysts of G. vanasi (Landsberg & Paperna, 1987) were found which became sporulated in tap-water within 24 hours. Another species in the gut of chubbyhead barb Barbus anoplus Weber harboured sporulated oöcysts in the faeces and in the intestinal epithelium. The latter species has been described as G. anopli n. sp.

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Garnia karyolytica n. sp. (Apicomplexa: Haemosporina: Garniidae), a blood parasite of the Brazilian lizard Thecodactylus rapicaudus (Squamata: Gekkonidae).

Development of meronts and gametocytes of Garnia karyolytica nov. sp., is described in erythrocytes of the neotropical forest gecko Thecodactylus rapicaudus from Pará State, north Brazil. Meronts are round to subspherical and predominantly polar in position: forms reaching 12.0 x 10.0 microns contain from 20-28 nuclei. Macrogametocytes and microgametocytes are predominantly elongate, lateral in the erythrocyte and average 16.6 x 6.3 microns and 15.25 x 6.24 microns respectively. Occasional spherical forms of both sexes occur in a polar or lateropolar position. All stages of development are devoid of malarial pigment. They have a progressively lytic effect on the host-cell nucleus, particularly the mature gametocytes, which enlarge and deform the erythrocyte. Possible vector(s) of garniid parasites are considered, and phlebotomine sandflies are high on the list of suspects.

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On Eimeria bragancaensis n. sp. (Apicomplexa: Eimeriidae) and tissue-cysts of an unidentified protozoan in the bat Peropteryx macrotis (Chiroptera: Emballonuridae) from Amazonian Brazil.

A description is given of the mature oocysts and endogenous stages of Eimeria bragancaensis n. sp., from the small intestine of the bat Peropteryx macrotis from north Brazil. Oocysts are spherical to subspherical, 15.9 x 14.6 microns (range 14-17 x 14-17.7). The wall is of two layers: an outer, thicker one which is of a brownish-yellow colour and striated (pitted), and an inner one which is thin and colourless. The outer layer is frequently lost. No micropyle or oocyst residuum present, but the oocyst usually contains one or two polar bodies. Sporocysts 8.4 x 5.3 microns (range 6.25-9 x 4-6) with inconspicuous Stieda and substiedal bodies. Endogenous stages are intracytoplasmic in the epithelial cells of the small intestine, above the host-cell nucleus. Stages of merogony and gametogony are described. Abundant tissue-cysts of an unidentified protozoan, containing from 1-4 zoites, were found in the parenchyma cells of the liver and, less frequently, in the lamina propria of the small intestine. Their possible nature is discussed.

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Plasmodium kentropyxi n.sp. (Apicomplexa: Haemosporina: Plasmodiidae) and a Plasmodium tropiduri-like parasite in the lizard Kentropyx calcarata (Lacertilia: Teiidae) in north Brazil.

Plasmodium kentropyxi n.sp. is described in the teiid lizard Kentropyx calcarata from north Brazil. Young asexual stages and gametocytes are at first polar in the erythrocyte but with elongation, move to a lateral position. Largest meronts seen contained from 30-40 nuclei and conspicuous greenish-black pigment granules located in a distinct vacuole. With growth the gametocytes eventually assume a smooth, curved cylindrical shape, with evenly rounded ends. Pigment is scattered or concentrated around a conspicuous vacuole which is slowly developed as the gametocytes mature. Mature male parasites measured 11.8 x 4.0 microns (9.6 x 4.2 - 13.2 x 3.6 microns), shape-index 2.9 (2.2 - 5.0), and females 13.5 x 4.5 microns (12.0 x 4.5 - 15.0 x 4.8 microns), shape-index 3.0 (2.2 - 3.8). Some larger meronts may slightly enlarge the erythrocyte, but most asexual stages and the mature gametocytes rarely do so. A second, P. tropiduri-like parasite encountered in K. calcarata possessed small rounded or fan-shaped meronts producing from 4-14 merozoites, and spherical to subspherical gametocytes of approximately 6.0 x 5.0 microns. The parasite was consistently polar in its position in the erythrocyte.

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Caryospora matatu n. sp., a new coccidian parasite (Apicomplexa: Eimeriidae) from the horned bush-viper, Atheris ceratophorus Werner, 1895 from Tanzania.

Caryospora matatu n. sp. is the first species of coccidia reported from the a horned bush viper, Atheris ceratophorus endemic to Tanzania. Oocysts are spherical or slightly subspherical, 19.8 (16-23) microns, a micropyle and an oocyst residuum are absent, a single polar granule is present. An oocyst wall is bilayered, approximately 1.5 microns thick, brownish and distinctly pitted. Sporocysts are ellipsoidal, 15.6 (12.5-17.0) x 10.1 (8-12) microns, tightly fitting an internal oocyst diameter, Stieda and substieda bodies are present. An experimental inoculation of SCID mice did not confirm a facultatively heteroxenous life cycle in the studied coccidium. Morphology and evolutionary history of Caryospora species from viperids is is reviewed and discussed.

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Two new species of Eimeria (Apicomplexa: Eimeriidae) from Asian geoemydid turtles Kachuga tentoria and Melanochelys trijuga (Testudines: Geoemydidae).

Coprological examination of ten Pink-ringed tent turtles Kachuga tentoria circumdata, recently imported from India, and three Burmese black turtles Melanochelys trijuga edeniana, imported from Myanmar, revealed the presence of two new species of Eimeria. Oocysts of Eimeria kachua n. sp. from K. t. circumdata are broadly oval to subspherical, 15.3 (13-18) x 13.9 (12-16) microm, with polar granule and subspherical oocyst residuum. Sporocyst elongatelly oval to spindle-shaped, 8.7 (7.5-10) x 4.9 (4-6) microm, with a knoblike Stieda body, covered with fine membranous cupola-like structures. Thin walled oocysts of Eimeria patta n. sp. from M. t. edeniana, have an irregular shape, influenced by the position of sporocysts, frequently with lobular irregularities, 12.6 (11-16) x 9.1 (7.5-12) microm. Sporocysts are oval to ellipsoidal, 5.8 (5-7) x 4.2 (3.5-5) microm.

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The phylogeny of Goussia and Choleoeimeria (Apicomplexa; Eimeriorina) and the evolution of excystation structures in coccidia.

The phylogenetic relationships of Goussia janae and Choleoeimeria sp. were analyzed using the small subunit ribosomal RNA gene (SSU rDNA). This is a first attempt to study the molecular phylogeny of coccidian genera parasitizing strictly poikilotherm hosts. The biliary Eimeria-like coccidia of reptiles classified into the genus Choleoeimeria form a sister clade to the family Eimeriidae, which confirms the separate generic status of the genus Choleoeimeria. The position of Goussia is less robustly resolved, since it forms a trichotomy with the Eimeriidae and Sarcocystidae, or alternatively constitutes the earliest branch of the coccidian lineage. Morphological similarities, namely the extracytoplasmic location of the endogenous stages, and the presence of sutures in the sporocyst wall are discussed in the context of the traditional classification of eimeriids. In contrast to the morphology-based systematics, the monophyly of Goussia and Choleoeimeria is not supported by the SSU rDNA data.

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Phylogenetic position of a renal coccidium of the European green frogs, 'Isospora' lieberkuehni Labbé, 1894 (Apicomplexa: Sarcocystidae) and its taxonomic implications.

'Isospora' lieberkuehni, an unusual isosporoid renal coccidium that parasitizes the European water frog was isolated from the edible frog, Rana kl. esculenta, in the Czech Republic. Sequencing of the small-subunit (SSU) rRNA gene showed that it belongs to the family Sarcocystidae, being closely related to a clade comprising members of the subfamily Toxoplasmatinae. The position within Sarcocystidae correlates with the mode of excystation via collapsible plates as postulated by previous authors. Phylogenetic, morphological and biological differences between 'Isospora' lieberkuehni and the other Stiedabody-lacking members of the genus Isospora justify separation of this coccidium on a generic level. Hyaloklossia Labbé, 1896 is the oldest available synonym and is herein re-erected. The original definition of the genus Hyaloklossia is emended based on recent observations.

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Ultrastructure of Goussia cruciata (Apicomplexa: Coccidia) infecting the liver of horse mackerel, Trachurus trachurus (L.), from Ibero-Atlantic waters.

The ultrastructure of developmental stages of Goussia cruciata and the pathology they cause in the liver of Trachurus trachurus (Teleostei: Carangidae) caught off the Galician (North-West Spain) and Portuguese North Atlantic coasts are described. Each oocyst contained four ellipsoidal sporocysts, with two sporozoites. The sporocyst wall consisted of a thick and dense inner layer with transverse striations and a multi-lamellated outer layer formed by parallel dense internal bands alternating with lighter areas. The lamellae formed filamentous extensions of the wall. The sporocyst wall striation period was smaller than that observed in G. clupearum, which has a similar habitat. The dehiscence suture, characteristic of the genus, was present in the sporocysts of G. cruciata. The sporocysts were arranged in a symmetrical and characteristic cross shape. A large number of sporocysts with sporozoites were observed in direct contact with host liver cells. No macroscopic lesions were observed. In heavily infected fish, aggregations of oocysts were often enveloped in a 'yellow body' composed of amylopectin granules derived from the parasite and necrotic or aggregated host cells. Degenerating parasites were frequently observed in liver tissue. Host inflammatory cells were accumulated near some oocysts. The ultrastructure of the parasite, together with its strict host specificity, confirmed G. cruciata as a separate and valid species.

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Some corrections in Haemogregarine (Apicomplexa: Protozoa) nomenclature.

The nomenclature of three genera in the family Haemogregarinidae (Haemogregarina, Karyolysus, and Hepatozoon) has been reviewed and the following new names are introduced to replace homonyms or for previously unnamed species: haemogregarina carlosi n. nom., in the erythrocytes of the lizard Lacerta ocellata; Haemogregarina tincae n. nom., in the stomach and intestine of the tench Tinca tinca; Hepatozoon insectivorae n. sp., in the leucocytes of the shrews Sorex araneus and Crocidura leucodon; Hepatozoon krampitzi n. sp., in the leucocytes of the vole Microtus oeconomus; Hepatozoon peromysci n. sp., in the leucocytes of the deermice Peromyscus boylii and P. truei gilberti; and Hepatozoon pallida (Pessoa et al., 1971) n. comb., in the erythrocytes of the snake Thamnodynastes pallidus nattereri.

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Transmission electron microscopy of intracellular sporozoites of Eimeria vermiformis (Apicomplexa, Eucoccidiida) in the mouse.

Sporozoites of Eimeria vermiformis from the mouse were first seen in the epithelial cells of villus tips and the crypts of Lieberkühn four hours after inoculation (HAI). They were always within a parasitophorous vacuole. By 12 HAI, most were in crypt epithelial cells between the basement membrane and host cell nucleus. The sporozoites in the villus tips had 26 subpellicular microtubules, two polar rings, two preconoidal rings, two refractile bodies surrounded by amylopectin-like granules, a lamellar Golgi apparatus, numerous micronemes, and rhoptries. The sporozoites in the crypt cells had fewer amylopectin-like granules, micronemes, and rhoptries. A nucleolus was visible, as were pieces broken off from the posterior refractile body. Later, the sporozoites folded over to become U-shaped; the infolded membranes fused; and then the inner membranes disappeared so that spherical meronts were formed. Folding sporozoites were first seen 16 HAI and persisted until 52 HAI.

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Transmission electron microscopy of meront development of Eimeria vermiformis Ernst, Chobotar and Hammond, 1971 (Apicomplexa, Eucoccidiorida) in the mouse, Mus musculus.

First and second generation meronts of Eimeria vermiformis developed in epithelial cells of the crypts of Lieberkühn. They were usually between the host cell nucleus and the basement membrane. Sporozoite organelles dedifferentiated with the first generation meront's development except for the refractile body and the apical complex, which persisted. After several nuclear divisions, the apical complex dedifferentiated further until only micronemes remained attached by a duct system to the plasmalemma. The form of the apical complex was highly variable. Sometimes the duct system was absent and the micronemes were attached directly to the plasmalemma or a dense material on it. Crescent body-like material was often present in the parasitophorous vacuole next to the microneme structure. The microneme structure was not present in second generation meronts but evaginations of the plasmalemma, cytoplasmic outpocketings, and cytoplasmic vesicles were associated with the round granular bodies in the parasitophorous vacuoles. During first generation merogenesis, invaginations from the parasitophorous vacuole formed channels into the meront along which merozoites budded. Micropores were often at the ends of these invaginations. These and other micropores of the meront had a dense U-shaped band for a collar while those of the merozoites had a collar with a double band of dense material that connected to the inner membrane. First generation merozoites budded randomly from the meront, resulting in a residual body that was usually in the middle of the parasitophorous vacuole. Second generation merozoites budded in one direction, resulting in a peripheral residual body and merozoites that were parallel in an oblong parasitophorous vacuole.

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Eimeria beldingii n. sp. and other Coccidia (Apicomplexa) of the ground squirrel Spermophilus beldingi.

Fecal samples of 36 ground squirrels, Spermophilus beldingi, from Tioga Pass (elev. ca. 3315 m) in the Sierra Nevada, California, yielded oocysts of Eimeria beckeri in nine squirrels, E. citelli in four squirrels, E. beldingii n. sp. in two squirrels, and degenerated, unidentifiable oocysts in ten squirrels. Eimeria beldingii n. sp. oocysts are ellipsoidal, 30-34 X 24-30 (mean 32 X 26) micron with a two-layered, rough, striated wall, without a micropyle or residuum, with polar granules; they contain ellipsoidal or ovoid sporocysts 11-15 X 9-12 (mean 13 X 10) micron with a Stieda body and residuum.

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Extraintestinal development of Caryospora simplex (Apicomplexa: Eimeriidae) in experimentally infected mice, Mus musculus.

Developmental stages of Caryospora simplex were found in connective tissue of the cheek, tongue, and nose of Swiss-Webster and C57 BL/6 mice (Mus musculus) from 8 through 70 days after oral inoculation with 50,000 or 250,000 oocysts, or 60,000 free sporocysts of the same species obtained from an Ottoman viper, Vipera xanthina xanthina. The earliest developmental stages were seen on day 8 post-inoculation (PI) and consisted of two types of meronts and gamonts (undifferentiated sexual stages). Gamonts, microgametocytes, macrogametes, and unsporulated oocysts were found on days 10 and 12 PI. Fully sporulated, thin-walled oocysts containing eight sporozoites surrounded by a thin sporocyst membrane were first seen 12 days PI. Monozoic cysts (caryocysts) were first seen 12 days PI and appeared fully viable throughout the duration of the study, 70 days PI. Four mice injected intra-peritoneally with 150,000 free sporozoites and killed 12 days PI contained unsporulated and sporulated oocysts in connective tissues of the cheek, tongue, and nose, suggesting that sporozoites may be carried to the site of infection via the lymphatic/circulatory system. Four cotton rats, Sigmodon hispidus, inoculated orally with 250,000 oocysts all had unsporulated and sporulated oocysts of C. simplex in connective tissue of the cheek, tongue, and nose when killed on day 12 PI, indicating extraintestinal development in the secondary host is not species specific. This is the first report of a heteroxenous coccidium with both asexual and sexual development in the primary (predator) and secondary (prey) hosts.

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