Iridovirus-like viruses in erythrocytes of lacertids from Portugal.

Icosahedral nucleo-cytoplasmic large DNA viruses (NCLDV)-like viruses, which forminclusions in the erythrocyte cytoplasm of reptiles, were previously presented as candidates for a new genus of the Iridoviridae family. The present work describes the distribution of infected lizard hosts and ultrastructural characteristics of the viral inclusions of NCLDV-like viruses from Portugal and adjacent locations in Spain. Giemsa-stained blood smears of 235 Lacerta schreiberi from Portugal and Spain, 571 Lacerta monticola from the mountain Serra da Estrela (Portugal), 794 Podarcis hispanica from several localities in Portugal and Spain, and 25 Lacerta dugesii from Madeira Island, were studied. Infection in L. schreiberi was only found in mountain populations, up to 30% in Serra da Estrela and 9-11% elsewhere. It was absent in lizards from lowlands. Prevalence of infection among L. monticola in Serra da Estrela was 10%; infected lizards were found during March to July and October but not in August and September. Infection in P. hispanica was below 3.3%. Only one infected specimen of L. dugesii was identified by light microscopy. Ultrastructural examination of infected samples revealed that the inclusions are virus assembly sites of icosahedral cytoplasmic iridovirus-like virions. Virions from different host species have different ultrastructural features and probably represent different related viruses.

Lankesterella poeppigii n. sp. (Apicomplexa, Lankesterellidae) from Bufo poeppigii (Tschudi, 1845) from Peru

Lankesterella poeppigii n. sp. is described from Bufo poeppigii (Tschudi, 1845) from Peru. Merogony and oogonyoccur in the capillary endothelium and the macrophages in the liver, spleen and kidneys. Meronts are oval, 25,2–29,4 x 15,7–16,8 ìm in size and yield 35–46 merozoites. Oocysts are 26,3–29,4 x 15,1–17,6 ìm in size; sporozoites 9,2-9,8 x 4,2–5,0 ìm in size, assemble in macrophages. Released 8,7–9,8 x 2,8–3,1 ìm sporozoites enter erythrocytes. L. poeppigii is compared with Lankesterella petiti Lainson & Paperna, 1995 infecting Bufomarinus (Linnaeus, 1758) in Brazil. The above mentioned specific characters, added to differences in hosts and geographical location warrant the description of Lankesterella poeppigii from B. poeppigii as a new species.

Lankesterella poeppigii n. sp. es descrita de Bufo poeppigii (Tschudi, 1845) del Perú. La merogonia y oogoniase producen en el endotelio capilar y los macrófagos en el hígado, el bazo y los riñones. Los esquizontes sonovalados, 25,2–29,4 x 15,7–16,8 micras de tamaño y producen 35–46 merozoitos. Los ooquistes miden 26,3–29,4x 15,1–17,6 micras de tamaño; esporozoitos, reunidos en los macrófagos, miden 9,2-9,8 x 4,2-5,0 micras detamaño. Liberados, los esporozoitos miden 8,7–9,8 x 2,8–3,1 micras y entran en los eritrocitos. Lankesterella poeppigii es comparada con L. petiti Lainson y Paperna, 1995, que infecta a Bufo marinus (Linnaeus, 1758) en Brasil. Los caracteres específicos citados, sumados a las diferencias entre los huespédes y en la localización geográfica justifican la clasificación de la Lankesterella de B. poeppigii como una nueva especie.

Haemosporidian infections in captive exotic glossy starling Lamprotornis chalybaeus in Hong Kong.

A greater blue-eared glossy starling Lamprotornis chalybaeus Ehrenburg from a large flight aviary in Hong Kong was found on post mortem to be infected with Plasmodium octamerium Manwell, 1968, Plasmodium cf. relictum (Grassi et Feletti, 1891) and Haemoproteus cf. pastoris Mello, 1935. Descriptions of their morphology are provided as none of the examined parasites fully concord with their type (or neotype) material descriptions. Plasmodium octamerium has been recorded in avian hosts from geographically distant locations, suggesting that infection in imported hosts may persist in a chronic state for a long period. This Plasmodium species as well as P. relictum are evidently not fastidious in choice of passeriform hosts and are transmitted by ubiquitous domestic mosquito vectors, apparently facilitating their proliferation among zoo and aviary inhabitants. The Haemoproteus infection appears to be conspecific with H. cf. pastoris reported from a myna (Acridotheres tristis) in Singapore. Mynas are also common in Hong Kong, which suggests a possible cross-transmission of infection between these two starlings.

Proliferative visceral Isospora (atoxoplasmosis) with morbid impact on the Israeli sparrow Passer domesticus biblicus Hartert, 1904.

House sparrows (Passer domesticus biblicus Hartert, 1904) caught in the Jordan valley, the coastal plain, and the desert region in Southern Israel were found massively infected with extraintestinal proliferative stages of Isospora, previously named Atoxoplasma. Infection coincided with Isospora spp. infections in the digestive tract. Prevalence of infection reached 70% among sparrows of all three regions; however, only in the Jordan valley did the severity of the sparrows compromised their survival. Healthy appearing captured birds showed symptoms of "going light" syndrome -- diarrhea, emaciation, and death. Birds succumbed within 48 h to 15 days after confinement to cages. Merozoites accumulated predominantly in the spleen but were rarely found in the peripheral blood. The parasite stages in the visceral leukocytes propagated by merogony and yielded single large waiting-stage merozoites. Visceral infections resulted in multifocal necrosis. Proliferative visceral Isospora infection (atoxoplasmosis) is one of the more severe causes of mortality among captive birds, free-ranging birds appear to coexist with the infection but succumb under capture stress.

Isospora carliae sp. n. (Apicomplexa: Eimeriidae) from the skink Carlia rhomboidalis (Peters) from Daintree Forest, North Queensland, Australia: description and fine-structural account of endogenous development.

Isospora carliae sp. n. is described from the blue-throated rainbow skink Carlia rhomboidalis (Peters), from Daintree Forest, North Queensland, Australia. Oocysts are ellipsoidal, 16.8-21.0 x 12.6-15.4 microm in size, with their two sporocysts, 9.0-14.0 x 7.0-9.24 microm in size, positioned along the wide axis. Sporozoites contain a distinct refractile body and are accompanied by a residuum. All endogenous development occurs within the host-cell nucleus. Nuclei are sometimes invaded by several merozoites, but only infections by a single parasite persist. Nuclei lodging meronts, mature microgamonts and premature macrogamonts have an elongate shape. Some meronts exhibit a membrane-bound cytoplasmic inclusion that contains many micronemes.

Leucocytozoonosis in the Israeli sparrow, Passer domesticus biblicus Hartert 1904.

Among the 91 house sparrows (Passer domesticus biblicus Hartert, 1904) examined and caught in the Jordan valley, Israel, 79% were found to be infected with Leucocytozoon fringillinarum Woodcock 1910. In the coastal plain of Israel (South of Tel Aviv), Leucocytozoon infection was found in only 3 out of 43 examined sparrows. In the birds examined, Leucocytozoon gametocytes were present, often in large numbers, in the circulating blood of the visceral organs, whereas they were only sporadic or even absent in the peripheral blood. Gametocytes were seen in the brain capillaries in only a few birds. Only one of the heavily infected sparrows was anemic. Leucocytozoon merozoites were present in the liver and kidneys in only a few infected birds. Merogonic infections did not induce any severe pathological changes, while the gametocyte congestion caused dilation of the blood vessels and sinuses. Tissue damage by the gametocyte parasitemia was most evident in the liver and kidneys. Leucocyte infiltration developed alongside the affected vessels; diffuse necrosis developed in the infiltrated areas. In the kidneys, many tubules were degenerated. Leucocytozoon gametocyte infection in sparrows is unique in that it appears to be confined, for most of its duration, to the visceral circulation, resulting in clinical consequences. Geographically, it is confined to habitats presumably supporting vectors.

The endogenous development, described by light and electron microscopy, of Eimeria jamescooki sp. n. (Apicomplexa: Eimeriidae) from the skink Cryptoblepharus virgatus.

Eimeria jamescooki sp. n. was recovered from the skink Cryptoblepharus virgatus (Garman) found on the grounds of James Cook University, Townsville (type locality), North Queensland, Australia. Oocysts were 17.5-25.0 (22.1 +/- 1.9) x 15-22.5 (17.7 +/- 1.6) microm and sporocysts 6.25-10.0 (7.9 +/- 1.15) x 3.75-6.25 (5.3 +/- 1.0) microm in size. Endogenous stages are described from histological material examined by light microscope and by transmission electron microscope. Both merogony stages and gamonts were found to develop in the cytoplasm of the anterior gut mucosal epithelium. Meront progeny were comprised of 10 to 21 merozoites. Premature macrogamonts were elongate; some host cells contained two elongate macrogamonts. Unique to the presently described species were the Golgi "plaques" and an enclosure of tubuli. Mature macrogamonts and young oocysts ranged in size from 14 x 7 to 21 x 11 microm and contained two types of wall-forming bodies, canaliculi and amylopectin granules. Differentiating microgamonts conformed in fine structure with that observed in other eimerians. Their sizes increased from 15.4 x 4.2 to 28 x 8.4 microm while dividing to over 70 nuclei, which formed a corresponding yield of microgametes.

Schizogonic stages of Haemoproteus from Wenyon's Baghdad sparrows are also found in Passer domesticus biblicus in Israel.

Schizont bodies reminiscent of those described by Wenyon from Baghdad sparrows were found in the liver and lungs of an Israeli house sparrow ( Passer domesticus biblicusHartert 1904) infected with Haemoproteus passeris Kruse, 1890. All observed schizonts were composed of packed assemblages of walled compartments, each holding a differentiating schizogonic body. The schizogonic bodies in the various compartments demonstrated sequential stages in the differentiation process from a compact multinucleate cytoplasmic mass to massive formation of multiple merozoites. Young non-differentiated schizont assemblages reached 0.2 x 0.25 mm in size and the fully differentiated ones, containing merozoites, were double or triple that. Among species of Haemopterus, division to compartmented and single-plasmodium schizonts could not be associated with any recognized generic or infrageneric division. Moreover, in some species of Haemoproteus, both types of schizogony co-existed.

Development of Hepatozoon caimani (Carini, 1909) Pess a, De Biasi & De Souza, 1972 in the Caiman Caiman c. crocodilus, the frog Rana catesbeiana and the mosquito Culex fatigans.

The sporogony of Hepatozoon caimani has been studied, by light microscopy, in the mosquito Culex fatigans fed on specimens of the caiman Caiman c. crocodilus showing gametocytes in their peripheral blood. Sporonts iniciate development in the space between the epithelium of the insect gut and the elastic membrane covering the haemocoele surface of the stomach. Sporulating oocysts are clustered on the gut, still invested by the gut surface membrane. Fully mature oocysts were first seen 21 days after the blood-meal. No sporogonic stages were found in some unidentified leeches fed on an infected caiman, up to 30 days following the blood-meal. When mosquitoes containing mature oocysts were fed to frogs (Leptodactylus fuscus and Rana catesbeiana), cysts containing cystozoites developed in the internal organs, principally the liver. Feeding these frogs to farm-bred caimans resulted in the appearance of gametocytes in their peripheral blood at some time between 59 and 79 days later, and the development of tissue cysts in the liver, spleen, lungs and kidneys. Transmission of the parasite was also obtained by feeding young caimans with infected mosquitoes and it is suggested that both methods occur in nature. The finding of similar cysts containing cystozoites in the semi-aquatic lizard Neusticurus bicarinatus, experimentally fed with infected C. fatigans, suggests that other secondary hosts may be involved.

Development of Hepatozoon caimani (Carini, 1909) Pessôa, De Biasi & De Souza, 1972 in the Caiman Caiman c. crocodilus, the frog Rana catesbeiana and the mosquito Culex fatigans

The sporogony of Hepatozoon caimani has been studied, by light microscopy, in the mosquito Culex fatigans fed on specimens of the caiman Caiman c. crocodilus showing gametocytes in their peripheral blood. Sporonts iniciate development in the space between the epithelium of the insect gut and the elastic membrane covering the haemocoele surface of the stomach. Sporulating oocysts are clustered on the gut, still invested by the gut surface membrane. Fully mature oocysts were first seen 21 days after the blood-meal. No sporogonic stages were found in some unidentified leeches fed on an infected caiman, up to 30 days following the blood-meal. When mosquitoes containing mature oocysts were fed to frogs (Leptodactylus fuscus and Rana catesbeiana), cysts containing cystozoites developed in the internal organs, principally the liver. Feeding these frogs to farm-bred caimans resulted in the appearance of gametocytes in their peripheral blood at some time between 59 and 79 days later, and the development of tissue cysts in the liver, spleen, lungs and kidneys. Transmission of the parasite was also obtained by feeding young caimans with infected mosquitoes and it is suggested that both methods occur in nature. The finding of similar cysts containing cystozoites in the semi-aquatic lizard Neusticurus bicarinatus, experimentally fed with infected C. fatigans, suggests that other secondary hosts may be involved

Experimental infection of lacertids with lizard erythrocytic viruses.

OBJECTIVE: Lizard erythrocytic viruses (LEVs) produce inclusions in the cytoplasm of erythrocytes, but their impact on the infected host is poorly understood. This work reports on an experimental study of the infection process in Lacerta monticola and Lacerta schreiberi from Serra da Estrela Mountain, Portugal. METHODS: A time sequence light microscope and transmission electron microscope (TEM) study of the infection process was performed in peripheral blood erythrocytes of experimentally infected lizards. Virions were searched for by TEM in visceral organs and bone marrow of the animals. RESULTS: Infection was usually restricted to erythrocytes, but occasionally became systemic and induced disease. In the first case, a prevalence of infected erythrocytes of up to 98% followed by recovery was observed. In the latter, infection spread to leukocytes, leading to the death of the infected animals. CONCLUSIONS: The potential of LEVs to induce systemic infections was demonstrated. Sequential TEM examination of LEV-infected cells is described for the first time, demonstrating features such as dense inclusions related to virus nucleoid formation, intranuclear virions, intermediate structures in virion capsid morphogenesis and virus release by budding.

Fine structure of erythrocytic stages of a Plasmodium tropiduri-like malaria parasite found in the lizard Kentropyx calcarata (Teiidae) from north Brazil.

The fine structure is described of the merogonic stages and gametocytes of a Plasmodium tropiduri Aragão et Neiva, 1909-like parasite infecting the teiid lizard Kentropyx calcarata Spix from North Brazil. The trophozoites are bordered by two membranes, and with growth a pellicle is formed by the addition of an inner, thick double layer and fragmented membrane. The same type of inner membrane occurs in the pellicle of the merozoites differentiating from the meronts. Merozoites contained a large electron-dense body, sometimes seen to be embraced by a tubular mitochondrion with a dense matrix. Micro- and macrogametocytes are bounded by a double membrane, closely apposed by the detached wall of the parasitophorous vacuole. Both contain osmiophilic bodies. The microgametocyte contains an electron-dense aggregate, and the macrogametocyte has a large mitochondrion and a complex of tubuli and cisternae. These features are compared with those described in other malarial parasites.

Fine structure of Sarcocystis singaporensis merogony stages preceding sarcocyst formation in the rat.

The fine structure of merogony stages of Sarcocystis singaporensis (Zaman and Colley, 1975) is described from experimentally infected laboratory rats, 10 days after being fed sporocysts obtained from naturally infected Python reticulatus from Singapore. Infection was shown to consist exclusively of S. singaporensis. Parasites developed in the endothelial cells of the lungs, brain, kidney and heart. Infection comprised meronts prior to division, dividing and divided meronts, and dispersed merozoites. Undivided meronts developed deep pellicular invaginations and extensions of the nucleus toward the cell boundary, seemingly to sustain metabolic exchange. The course of merogonous development was the same in all organs. Hypertrophy of the endothelium induced by invading merozoites appeared to lead to obstruction of the capillary lumen.

Plasmodium neusticuri n. sp. (Apicomplexa: Plasmodiidae), a parasite of the lizard Neusticurus bicarinatus (Lacertilia: Teiidae) in Amazonian Brazil

Plasmodium neusticuri n.sp. (Apicomplexa: Plasmodiidae) is described in the blood of the lizard Neusticurus bicarinatus (Lacertilia: Teiidae) from Amazonian Brazil. All stages of the parasite occupy a polar position in the erythrocyte and produce no enlargement of the host cell and little or no distortion of its nucleus. Trophozoites commonly display a variable number of long filopodia, and quickly produce a small number of dark, yellow-green pigment granules. Meronts are smaller than the host-cell nucleus. They are broadly ellipsoidal to spherical, produce 4-8 (usually 6) merozoites disposed in an irregular manner, and have some 5-10 pigment granules forming a loose clump. Macrogametocytes and microgametocytes are broadly ellipsoidal, frequently "kidney-shaped", rarely spherical.They are generally smaller than the host-cell nucleus, averaging 5.8 x 4.1 mum and 5.8 x 4.4 mum, respectively. Both usually exhibit a conspicuous cytoplasmic vacuole, and their pigment granules are more abundant and scattered than those of the meronts. Rare examples of extraerythrocytic meronts were encountered in monocytes. The vector of the parasite is at present unknown.

A new filaria of a lizard transmitted by sandflies

A description is given of Madathamugadia wanjii n. sp., a Splendidofilariinae parasite of the gecko Ptyodactylus hasselquistii, on the west bank of the River Jordan and of its life cycle in Phlebotomus duboscqi. The new species is close to M. ivaschkini (Annaev, 1976) n. comb., of Turkmenistan, wich is also transmitted by sandflies (Reznik, 1982). The genus Madathamugadia is now comprised of four species, two from Madagascar and two from the Mediterranean sub-region; it differs from the genus Thamugadia by the presence of a double row of papillae anterior to the cloaca of the male. The larval characters of Splendidofilarinae of lizards confirm the affinity of these parasites to the Splendidofilarinae of birds (Chandlerella and Splendidofilaria); the first group could have arisen from the second by "captures" wich could have occurred in several places