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In this study, we provide phylogenetic and biogeographic evidence that the Trypanosoma cruzi lineages T. cruzi I (TCI) and T. cruzi IIa (TCIIa) circulate amongst non-human primates in Brazilian Amazonia, and are transmitted by Rhodnius species in overlapping arboreal transmission cycles, sporadically infecting humans. TCI presented higher prevalence rates, and no lineages other than TCI and TCIIa were found in this study in wild monkeys and Rhodnius from the Amazonian region. We characterised TCI and TCIIa from wild primates (16 TCI and five TCIIa), Rhodnius spp. (13 TCI and nine TCIIa), and humans with Chagas disease associated with oral transmission (14 TCI and five TCIIa) in Brazilian Amazonia. To our knowledge, TCIIa had not been associated with wild monkeys until now. Polymorphisms of ssrDNA, cytochrome b gene sequences and randomly amplified polymorphic DNA (RAPD) patterns clearly separated TCIIa from TCIIb-e and TCI lineages, and disclosed small intra-lineage polymorphisms amongst isolates from Amazonia. These data are important in understanding the complexity of the transmission cycles, genetic structure, and evolutionary history of T. cruzi populations circulating in Amazonia, and they contribute to both the unravelling of human infection routes and the pathological peculiarities of Chagas disease in this region.

Parasites of wild primates are important for conservation biology and human health due to their high potential to infect humans. In the Amazon region, non-human primates are commonly infected by Trypanosoma cruzi and T. rangeli, which are also infective to man and several mammals. This is the first survey of trypanosomiasis in a critically endangered species of tamarin, Saguinus bicolor (Callitrichidae), from the Brazilian Amazon Rainforest. Of the 96 free-ranging specimens of S. bicolor examined 45 (46.8%) yielded blood smears positive for trypanosomes. T. rangeli was detected in blood smears of 38 monkeys (39.6%) whereas T. cruzi was never detected. Seven animals (7.3%) presented trypanosomes of the subgenus Megatrypanum. Hemocultures detected 84 positive tamarins (87.5%). Seventy-two of 84 (85.7%) were morphologically diagnosed as T. rangeli and 3 (3.1%) as T. cruzi. Nine tamarins (9.4%) yielded mixed cultures of these two species, which after successive passages generated six cultures exclusively of T. cruzi and two of T. rangeli, with only one culture remaining mixed. Of the 72 cultures positive for T. rangeli, 62 remained as established cultures and were genotyped: 8 were assigned to phylogenetic lineage A (12.9%) and 54 to lineage B (87.1%). Ten established cultures of T. cruzi were genotyped as TCI lineage (100%). Transmission of both trypanosome species, their potential risk to this endangered species and the role of wild primates as reservoirs for trypanosomes infective to humans are discussed.

Forty-six species of Lutzomyia and one species of Brumptomyia were identified among 20,008 sand flies collected in central Amapá. L. squamiventris maripaensis, L. infraspinosa, L. umbratilis, and L. ubiquitalis accounted for 66% of the specimens caught in light traps, and L. umbratilis was the commonest of the 16 species found on tree bases. Seven species of Lutzomyia including L. umbratilis were collected in a plantation of Caribbean pine. Sixty out of 511 female sand flies dissected were positive for flagellates. Among the sand flies from which Leishmania was isolated, promastigotes were observed in the salivary glands and foregut of 13 out of 21 females scored as having very heavy infections in the remainder of the gut, reinforcing the idea that salivary gland invasion may be part of the normal life cycle of Leishmania in nature. Salivary gland infections were detected in specimens of L. umbratilis, L. whitmani and L. spathotrichia. Parasites isolated from L. umbratilis, L. whitmani and also from one specimen of L. dendrophyla containing the remains of a bloodmeal, were compatible with Le. guyanensis by morphology and behaviour in hamsters.

Isolates of Paracoccidioides brasiliensis may vary in virulence according to time of in vitro subcultivation. The present study compared the morphology and pathogenicity to hamsters of two P. brasiliensis isolates: one obtained from human lesions and maintained in the laboratory for several years (Pb-18) and the other isolate recovered from hamsters inoculated with organ homogenates from armadillos (Pb-T). The microscopic morphology of Pb-18 and Pb-T showed yeast cells with similar diameter. However, Pb-T produced a significantly higher number of buds per mother cell than Pb-18. Besides, the mycelial form of Pb-T developed abundant sporulation during 8 weeks of culture which was absent in the Pb-18 isolate. Virulence studies demonstrated that mortality rates, antibody levels, fungal load and extent of lesions in the organs were significantly higher in animals infected with Pb-T. The results demonstrated that Pb-T recently isolated from an animal was more virulent than Pb-18. These differences between the two P. brasillensis isolates may be indicators of virulence attenuation in this fungal species.

Traditional molecular and biochemical methods, such as schizodeme analysis, karyotyping, DNA fingerprinting, and enzyme electrophoretic profiles, have shown a large variability among Trypanosoma cruzi isolates. In contrast to those results, polymerase chain reaction (PCR) amplification of sequences from the 24S alpha ribosomal RNA gene and from the mini-exon gene nontranscribed spacer indicated a dimorphism among T. cruzi isolates, which enabled the definition of two major parasite lineages. In the present study, 86 T. cruzi field stocks (68 isolated from humans with defined presentations of Chagas' disease and 18 from triatomines) derived from four Brazilian geographic areas were typed by the PCR assay based on the DNA sequences of the mini-exon and 24S alpha rRNA genes. These stocks were ordered into the two major T. cruzi lineages. Lineage 1 was associated mainly with human isolates and lineage 2 with the sylvatic cycle of the parasite.

A large number of Endotrypanum stocks (representing an heterogeneous population of strains) have been screened against a panel of monoclonal antibodies (MAbs) derived for selected species of Endotrypanum or Leishmania, to see whether this approach could be used to group/differentiate further among these parasites. Using different immunological assay systems, MAbs considered specific for the genus Endotrypanum (E-24, CXXX-3G5-F12) or strain M6159 of E. schaudinni (E-2, CXIV-3C7-F5) reacted variably according to the test used but in the ELISA or immunofluorescence assay both reacted with all the strains tested. Analyses using these MAbs showed antigenic diversity occurring among the Endotrypanum strains, but no qualitative or quantitative reactivity pattern could be consistently related to parasite origin (i.e., host species involved) or geographic area of isolation. Western blot analyses of the parasites showed that these MAbs recognized multiple components. Differences existed either in the epitope density or molecular forms associated with the antigenic determinants and therefore allowed the assignment of the strains to specific antigenic groups. Using immunofluorescence or ELISA assay, clone E-24 produced reaction with L. equatorensis (which is a parasite of sloth and rodent), but not with other trypanosomatids examined. Interestingly, the latter parasite and the Endotrypanum strains crossreacted with a number of MAbs that were produced against members of the L. major-L. tropica complex.

We report the detection, isolation and description of Trypanosoma (Megatrypanum) lainsoni n. sp. from a caviomorph rodent, Mesomys hispidus (Rodentia: Echimyidae), obtained in the Rio Negro region of the state of Amazonas, in northern BRA. Laboratory-bred white mice (Mus musculus) and rats (Rattus rattus) were inoculated with large numbers of culture forms by intraperitoneal route, and trypomastigotes appeared in their blood 3-8 days post-inoculation. One single epimastigote was also found in Mus musculus. Similar attempts to infect Rattus norvegicus, hamsters (Mesocricetus auratus), the opossum Didelphis marsupialis, the anteater Tamandua tetradactyla and triatomine bugs were unsuccessful, following six months of observations and microscopic examinations of blood films and blood cultures. As we have found no previous record of a Trypanosoma (Megatrypanum) species naturally infecting a member of the family Echimyidae, or any other caviomorph rodent, we conclude that this is the first time such an infection has been reported. The new species is unusual in the subgenus for its infectivity to laboratory mice. © 2013 R.D. Naiff et al., published by EDP Sciences.

0. Fernandesi*3, S. S. Santos’, E. CupoUlo’, B. Mendonga’, R. Derre3, A. C. V. Junqueira’, L. C. Santos’, N. R. Sturm4, R. l3. NaifP, T. V. Barret’, Il. A. Campbell4 and J. R. Coura’ ‘Department of Tropical Medicine and 2Department of Imw~nunology, Instiwo OswaEdo Cruz, Fiocruz, Rio de Janeiro, Brazil; 3DepaPtment of Pathology, Universidade do Estado do Rio de Janeiro, Rio de Janeiro, Brazil; 4Department of Microbiology, InwunoZogy and MolecuZar Genetics, Uv 51nstituto National de Pesquisas da AmazBnia, INPA, Manaus, Brazil Keywords:Americantrypanosomiasis, Chagasdisease

Ninety-four leishmanial isolates from the Brazilian Amazon Region (Amapa, Amazonas, Para, and Rondonia) were identified and classified using specific monoclonal antibodies and an indirect radioimmunoassay (serodeme analysis); eighty-two were also characterized by enzyme electrophoresis (zymodeme analysis), the results of which were subjected to a numerical phenetic analysis. Six isolates from humans (3), Didelphis marsupialis (1), Lutzomyia olmeca nociva (1), and Lu. reducta (1) showed reactivity patterns and isoenzyme profiles similar to those obtained with the Leishmania amazonensis reference strains, and were identified as this species. Eighty-six stocks were classified as members of the L. braziliensis complex; of these, 61 were L. guyanensis or variants, which presented three serodeme subtypes, but whose isoenzyme profiles were all similar to the reference strain. A total of 15 isolates were distinguished as L. braziliensis or variants and were classified into five serodeme subtypes. The isolate from Psychodopugus davisi appeared, from the numerical analysis, to be a distinct parasite species. Ten isolates showed reactivity patterns and isoenzyme profiles similar to those obtained with the L. naiffi reference strain. A parasite isolated from Ps. claustrei appeared to be different from all reference strains by both techniques, and was classified as probably being a new species. The importance of these results with respect to the taxonomic status of the New World Leishmania, and their implications for both clinical and epidemiologic data are discussed.

A new species of trypanosome, Trypanosoma (Megatrypanum) peba, is described from the peripheral blood of the armadillo Euphractus sexcinctus setosus from Bahia State, Brazil. Ten out of 29 specimens of the armadillo Dasypus novemcinctus from Pará State were found to have trypanosomes, including epimastigote forms, in impression smears of subcutaneous lymph nodes. The trypanosomes from D. novemcinctus are illustrated and were identified a idenrified as belonging to the subgenus Megatrypanum on the basis of their general appearance, although they failed to multiply is blood-agar culture medium and no bloodstream forms were seen. This is the first published record of trypanosomes of this subgenus from armdillos and the first demonstration of epimastigote trypanosomes in the mammalian host other than in the bloodstream, or in the anal glands of opssums.

Minist?rio da Sa?de. Secretaria de Vigil?ncia em Sa?de. Instituto Evandro Chagas, Bel?m, PA, Brasil. Instituto Nacional de Pesquisa da Amaz?nia. Manaus, AM, Brasil. Hebrew University of Jerusalen. Faculty of Agricultural, Food and Environmental Quality Sciences. Department of Animal Sciences. Rehovot, Israel. 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.

Trypanosoma cruzi is currently classified into 2 major phylogenetic lineages, T. cruzi I and II, that correlate with the formerly described zymodeme 1 and 2, respectively. Another isoenzymic group (zymodeme 3–Z3) was also described. In this study, we analysed the genetic diversity among Z3 isolates of the Brazilian Amazon by restriction fragment length polymorphism of the intergenic transcribed spacers (ITSs) of the ribosomal RNA cistron and the size of the divergent domain D7 of the 24Sα rRNA gene. DNAs from 12 T. cruzi Z3 isolates obtained from humans (2), Panstrongylus geniculatus (1), and Rhodnius brethesi (9) were submitted to PCR amplification of the ITSs plus the 5·8S rDNA. The PCR products were digested with 4 distinct endonucleases and the profiles analysed by a numerical methodology. The phenetic dendrogram revealed a clear dichotomy in the Z3 group, defining 2 groups that were named Z3-A and Z3-B. Dimorphism was also found in the band sizes of the amplified D7 divergent domain of the 24Sα rDNA, which showed a perfect correlation with the ITSs clustering. The organization of the ribosomal cistron was investigated by Southern blotting and shown to be conserved in the genome of the 2 Z3 groups. This study shows that the rDNA cistron allows the definition of 2 distinct subclusters in Z3 isolates.

The oocyst is described of Eimeria peltocephali n.sp. from faeces of the freshwater turtle Peltocephalus dumerilianus from Barcelos, State of Amazonas, Brazil. Sporulation is exogenous and fully developed oocysts are elongate, ellipsoidal or cylindrical, frequently curved to a banana-shape, 54.4 x 19.1 (37.5-68.7 x 18.7-20.0 microns), shape-index 2.8 (1.8-3.9). The oocyst wall is a single thin, colourless layer about 1 micron thick, with no micropyle. There is a bulky oocyst residuum, at first spherical to ellipsoidal, 19 x 16 (16.2-26.2 x 16-21.5 microns), but becoming dispersed on maturation. There are no polar bodies. The sporocysts, 19.1 x 6.8 (17.5-21.2 x 6.2-7.5 microns), shape-index 2.8 (2.3-3.2), are usually disposed in pairs at each end of the oocyst, and bear an inconspicuous Stieda body in the form of a flat cap. The sporozoites are elongate and slightly curved around the residuum. No refractile bodies were seen. Eimeria molossi n.sp., is described from the molossid bat Molossus ater. Sporulation is exogenous and the mature oocysts are predominantly broadly ellipsoidal, 23.4 x 17.5 (18-30 x 15-22.5 microns), shape-index 1.3 (1-1.6). The oocyst wall is about 2 microns thick, and of three layers: an inner thin, colourless one and two outer layers which are thicker, yellowish-brown, prominently striated and in close apposition. There is no micropyle or oocyst residuum, but one and occasionally two polar bodies are usually present. Sporocysts are ellipsoidal, 10.2 x 7.5 (10-12.5 x 7.5 microns), shape-index 1.4 (1.3-1.7) with an inconspicuous Stieda body. Endogenous stages are described in the epithelial cells of the small intestine.

It is the general opinion that the haemoproteid blood parasites of chelonians belong to the genus Haemoproteus . Different specific names have long been assigned to this parasite in birds, but some past authorities have accepted only a single species, H. metchnikovi , for all those haemoproteids recorded in a wide range of chelonian genera throughout the world. In the present study a comparison of one such organism in the tortoise Geochelone denticulata with another in the river turtle Peltocephalus dumerilianus , from Amazonian Brazil, has revealed clear morphological differences. These distinguish the parasites from each other, H. metchnikovi and the other named species of chelonian Haemoproteus for which adequate descriptions are available. We have assigned to them the names Haemoproteus geochelonis n.sp. and Haemoproteus peltocephali n.sp.

Adults of Rhodnius prolixus, R. pictipes and Panstrongylus geniculatus found in houses in Manaus have high rates of Trypanosoma cruzi infection. Rhodnius spp. were found throughout the year, but males of P. geniculatus were significantly more frequent in the dry season.

SUMMARYIn this study, we have analysed enzyme polymorphism among a group of protozoan parasites of the genus Endotrypanum (Kinetoplastida: Trypanosomatidae). Seventeen stocks of Endotrypanum spp. isolated from sloths (Choloepus didactylus and C. juruanus) in the Amazon Region of Brazil were analysed by enzyme electrophoresis, and their electromorphic profiles were compared with reference strains reported previously. The 16 enzymic loci were analysed, and the strains were classified into zymodemes, each representing parasites with unique enzyme profiles. Each zymodeme was considered as an elementary taxon, and using numerical analyses (cladistic, agglomerative hierarchical and ordination techniques) the genus was shown to be monophyletic and the 12 zymodemes characterized could be divided into 3 groups (A, B, C). The heterogeneous population (which may represent a complex of parasite species or strains variants) showed, however, no correlation with the origin (i.e. host species involved or geographic area of isolation) of Endotrypanum stocks. Eight isolates of Endotrypanum sp. from Rondonia State (Brazil) and a parasite strain from Panama were clustered together into a zymodeme, which was phenetically closely related to the E. monterogeii from Costa Rica. The data indicate that E. schaudinni is a species complex.

Vinte e oito amostras de Histoplasma capsulatum foram obtidas de oito especies de mamiferos silvestres nos Estados do Amazonas, Para e Rondonia. Os isolamentos foram feitos mediante inoculacao de amostras trituradas de figado e baco em hamsters por via intradermica e intraperitoneal. O diagnostico micologico nos hamsters que apresentaram lesoes foi confirmado por histopatologia e cultivo em meio dextrose-agar de Sabouraud. Os hamsters infectados desenvolveram sinais de doenca apos dois a nove meses; todos apresentaram lesoes disseminadas nas visceras e a maioria apresentou tambem lesoes cutâneas nos locais da inoculacao. Nenhum dos hamsters inoculados com material de pele dos hospedeiros originais desenvolveu histoplasmose, e H. capsulatum nao foi detectado nos exames histopatologicos dos animais silvestres. A prevalencia de infeccao foi consideravelmente mais alta entre femeas, tanto para Didelphis marsupialis como para o total de animais silvestres examinados (479). Propoe-se que as infeccoes detectadas em animais arboricolas podem ser explicadas pelo transporte de esporos do fungo em correntezas convectivas dentro de arvores ocas que tenham uma abertura ao nivel do solo e outra a nivel proximo das copas.

Submitted by Guilherme Lemeszenski (guilherme@nead.unesp.br) on 2013-08-22T19:04:25Z No. of bitstreams: 1 S0036-46651995000100007.pdf: 897597 bytes, checksum: 5cd06ce16898f7704f543141f811f999 (MD5) Made available in DSpace on 2013-08-22T19:04:25Z (GMT). No. of bitstreams: 1 S0036-46651995000100007.pdf: 897597 bytes, checksum: 5cd06ce16898f7704f543141f811f999 (MD5) Previous issue date: 1995-02-01 Made available in DSpace on 2013-09-30T20:07:21Z (GMT). No. of bitstreams: 2 S0036-46651995000100007.pdf: 897597 bytes, checksum: 5cd06ce16898f7704f543141f811f999 (MD5) S0036-46651995000100007.pdf.txt: 7 bytes, checksum: 212b0306580d4f0044d18f9a3edcc832 (MD5) Previous issue date: 1995-02-01 Submitted by Vitor Silverio Rodrigues (vitorsrodrigues@reitoria.unesp.br) on 2014-05-20T15:17:13Z No. of bitstreams: 2 S0036-46651995000100007.pdf: 897597 bytes, checksum: 5cd06ce16898f7704f543141f811f999 (MD5) S0036-46651995000100007.pdf.txt: 7 bytes, checksum: 212b0306580d4f0044d18f9a3edcc832 (MD5) Made available in DSpace on 2014-05-20T15:17:13Z (GMT). No. of bitstreams: 2 S0036-46651995000100007.pdf: 897597 bytes, checksum: 5cd06ce16898f7704f543141f811f999 (MD5) S0036-46651995000100007.pdf.txt: 7 bytes, checksum: 212b0306580d4f0044d18f9a3edcc832 (MD5) Previous issue date: 1995-02-01 Amostra de Paracoccidioides brasiliensis isolada de vísceras (baço e fígado) de um tatu (Dasipus novencinctus) foi estudada do ponto de vista micológico e imunoquímico. O tatu havia sido capturado em área da usina hidroelétrica de Tucuruí (Estado do Pará). Este já havia sido considerado como reservatório enzoótico do Paracoccidioides brasiliensis naquela região. Esta amostra, conservada na Micoteca do Instituto de Medicina Tropical de São Paulo sob o número 135, apresenta todas as características de Paracoccidioides brasiliensis, com elevado poder antigênico e baixa virulência para cobaios e ratos Wistar. A demonstração do exo-antígeno específico do P. brasiliensis, representado pela glicoproteína de peso molecular 43 kDa, foi evidente através das técnicas de Imunodifusão Dupla, Imunoeletroforese, SDS-PAGE e Imunoblotting. A sample of P. brasiliensis isolated from the spleen and the liver of an armadillo (Dasipus novencinctus) has been analysed under a mycological and immunochemical viewpoint. The armadillo was captured in an area of Tucuruí (State of Pará, Brazil), the animal being already established as an enzootic reservoir of P. brasiliensis at that region of the country. This sample maintained in the fungal collection of the Tropical Medicine Institute of São Paulo (Brazil) numbered 135, has got all the characteristics of P. brasiliensis, with a strong antigenic power and low virulence for guinea-pigs and Wistar rats. The specific exoantigen of P. brasiliensis - the glycoprotein with a molecular weight of 43 kDa - was easily demonstrated with double immunodiffusion, immunoelectrophoresis, SDS-PAGE and immunobloting techniques. Instituto de Medicina Tropical de São Paulo Laboratório de Micologia Médica USP Faculdade de Medicina Hospital das Clínicas Instituto Adolfo Lutz Seção de Micologia Instituto Nacional de Pesquisas da Amazônia UNESP Faculdade de Medicina de Botucatu UNESP Faculdade de Medicina de Botucatu

Como parte do programa do laboratorio de Parasitología do INPA, na procura dos vetores e reservatórios da Leishmaniose, foi realizado durante o periodo de 1978 até 1981, um levantamento da fauna flebotomínica, em oito localidades da Rodovia BR-319 (Manaus-Humaita). Capturamos 57 espécies: 44 do gênero Lutzomyiae 13 de Psychodopygus. Os métodos de coleta usados foram: capturas nos troncos das árvores, isca humana e armadilhas luminosas, o método mais eficiente foi o da armadilha luminosa colocada a 6 metros de altura na floresta. As espécies mais comuns foram L. antunesi, P. chagasie P. ayrozai.Dados sobre a distribuição geográfica das espécies coletadas e a distribuição por coletas são também assinalados. As part of our ongoing program, searching for the vectors and reservoirs of Leishmaniaparasites, we conducted a survey of the sandfly fauna along the BR-319 (Manaus-Porto Velho) highway. Fifty seven species of sandflies were captured; 44 in the genus Lutzomyia and 13 Psychodopygus.Of the collecting methods utilized, CDC traps set 6m above the forest floor proved to be the most effective, and the most frequently captured species were L. antunesi, P. chagasiand P. ayrozai.We also present the geographical distribution of the different species collected.

Leishmania naiffi was isolated from 10 out of 64 armadillos (Dasypus novemcinctus) examined in Amazonas, Pará and Rondônia States in the Brazilian Amazon Region. The isolates were obtained in culture from samples of liver (3), spleen (3), lymph nodes (2), skin (1) and blood (1) from the infected animals. Heavy infections with the same parasite were detected for the first time in Psychodopygus squamiventris, a common man-biting phlebotomine, in Amazonas and Pará. A new case of cutaneous leishmaniasis caused by L. naiffi is described from the Manaus area, making a total of three known cases of human infection by this parasite.

A total of 46 Trypanosoma rangeli stocks were isolated from naturally infected mammals and triatomine vectors. Twenty-two stocks were from the common opossum (Di- delphis marsupialis), one from the brown "4-eyed" opossum (Metachirus nudicaudatus), one from the anteater (Tamandua tetradactyla), one from the coati (Nasua nasua), seven from Rhodnius pictipes and 14 from Rhodnius robustus. Two stocks were also isolated from recently fed sandflies (Lutzomyia sp., Shannoni group). The stocks were identified as T. rangeli on the basis of natural or experimental salivary gland infections in Rhodnius, inoc- ulative (anterior station) transmission to mice, morphological parameters in parasitemic mice and com pari sons of isozyme profiles with a known stock of T. range li isolated from mano Three other trypanosome stocks from D. marsupialis, T. tetradactyla and the three-toed sloth (Bradypus tridactylus) were morphologically similar to T. range li in culture but had quite different isozyme profiles and were not identified. It is concluded that T. rangeli is widely distributed in Amazonas, Para and Rondonia States of Brazil, and probably extends into other regions where R. pictipes and R. robustus are known to occur. R. pictipes is light-attracted into houses and occasionally transmits Chagas' disease to mano It is likely that T. range li is also occasionally transmitted to man in the Amazon basin.

Flagellate infections were found in 1,063 of 18,895 sand flies collected in the states of Amazonas, Pará, Rondonia and Acre, Brazil. Infection rates were 13.4% (species group Shannoni); 7.5% (subgenus Nyssomyia); 6.7% (subgenus Lutzomyia series Cruciata); 0.5% (genus Psychodopygus) and 3.1% for other sand flies (various subgenera). Leishmania braziliensis guyanensis and L. mexicana amazonensis were isolated, respectively, from the known vectors, Lutzomyia umbratilis and L. flaviscutellata. Single stocks of L. braziliensis-like and L. mexicana-like organisms were isolated, respectively, from L. whitmani and L. yuilli. Thirty-eight flagellate stocks, isolated by direct culture from sand flies were characterized in detail by morphology in culture, behavior in hamsters and mice and by enzyme profiles. Sixteen stocks from Lutzomyia sp. (Shannoni group) were identified as Endotrypanum schaudinni; 8 stocks from Lutzomyia sp. (Shannoni group) were identified as Endotrypanum sp.; 7 stocks from Psychodopygus ayrozai and P. paraensis were identified as Leishmania sp. previously isolated from the armadillo, Dasypus novemcinctus; 2 stocks of Trypanosoma rangeli were isolated from recently fed Lutzomyia sp. (Shannoni group) sand flies; the remaining 5 stocks from L. umbratilis and L. yuilli could not be identified. Observations suggested that Shannoni group sand flies were the natural vectors of Endotrypanum. Leishmania sp. infections in the man-biting flies P. ayrozai and P. paraensis were restricted to the midgut and associated with recent bloodmeals. Unidentified flagellates in L. umbratilis and L. yuilli were distributed throughout the digestive tract with no trace of bloodmeals.

Cistos de Besnoitia foram encontrados nos músculos e vísceras de quinze exemplares de Didelphis marsupialis de um total de duzentos e vinte e quatro examinados. É a primeira vez que este protozoário e isolado de animais infectados naturalmente no Brasil. A transmissão experimental foi feita para animais de laboratório pela inoculação de triturados de tecidos e cistos.

A member of the Lutzomyia flaviscutellata complex from Rondônia and southern Amazonas States, Brazil, is so close to the Venezuelan Lutzomyia olmeca recuta Feliciangeli et al., 1988, that it is regarded as belonging to the same species. Since this phlebotomine co-extis with L. olmeca nociva in Brazil, the subspecific status of the former is untenable and is rased to specific rank, as Lutzomyia reducta. The Brazilian material is described and illustrated, and compared with specimens of L. o. nociva and L. flaviscutellata from the same area. Keys to the known taxa of the flaviscutellata complex are presented. Leishmania amazonensis was isolated from one heavily infected specimen of L. reducta, making this the third species of the flaviscutellata complex to be implicated as a vector of this parasite in Brazil. The relative abundance of the three sympatric flaviscutellata complex species varies locally and appears to be related to soil drainage. L. reducta constituted about 25% if all phlebotomines captured in Disney traps at poorly drained and well drained site, but appears not to coloniza areas subject to periodic flooding. L. olmeca nociva was restricted to poorly drained areas not subject to flooding, whereas L. flaviscutellata was ubiquitous L. reducta has never been detected north of the Amazon river in Brazil, but absence of recosrds from western and northwestern Amazonas State may reflect lack of collecting in these areas.

Of 151 Trypanosoma cruzi stocks from 117 different individual hosts collected in the states of Amazonas and Rondonia, 147 from 113 hosts were identified as zymodeme 1 (Z1). These included T. cruzi stocks from three marsupial species, two rodent species and three triatomine species although most were from the common opossum, Didelphis marsupialis. One T. cruzi stock from Rhodnius robustus was identified as Z1 with a Z3 PGM character, one from Sciurus sp. as Z3 and two from Monodelphis brevicaudata and Panstrongylus geniculatus as Z3 with a Z1 ASAT character. The ways in which stocks were isolated and grown up in vitro did not influence isozyme profile. These results support earlier evidence from Para State that Z2 is absent from the Amazon basin and that the distribution of T. cruzi zymodemes in this region is quite different to that in endemic areas on the south of the continent.

Minist?rio da Sa?de. Funda??o Servi?os de Sa?de P?blica. Instituto Evandro Chagas. Bel?m, PA, Brasil. Instituto Nacional de Pesquisas da Amaz?nia. Manaus, AM, Brasil. Instituto Nacional de Pesquisas da Amaz?nia. Manaus, AM, Brasil. Minist?rio da Sa?de. Funda??o Servi?os de Sa?de P?blica. Instituto Evandro Chagas. Bel?m, PA, Brasil. Eimeria trichechi n.sp. is described and figured from the manatee, Trichechus inunguis, from Amazonas State, north Brazil. Undifferentiated oocysts are passed in the faeces and complete their sporulation in ~36h at 24 to 24?C. Oocysts are spherical, with a mean size of 13.40 X 13.30 ?m. The wall is uni-layered, smooth, colourless and ~0.5 ?m thick; there is no micropyle, oocyst residuum or polar body. Sporocysts are ellipsoidal, with a men size of 8.65 X 4.62 ?m. The sporozoites are longer than the sporocyst and recurved at their ends: there is a sporocysts residuum of fine granules, but no visible steida body.

From a total of 224 Didelphis marsupialis examined, in 15 were found cysts of Besnoitia in muscles and viscera. It's the first time that this protozoan is isolated from naturally infected animals in Brazil. The experimental transmission to laboratory animals was done by the inoculum of tissue and cysts triturated.

Twenty one opossums (Didelphis marsupialis) from disturbed primary forest in the vicinity of Manaus, and seven from an isolated island of secondary vegetation within the city of Manaus, were examined for the presence of Leishmania. Of the opossums from the primary forests, 13 (61.9%) were found to be positive for Leishmania braziliensis guyanenesis. One additional opossum was found to be positive for Le. mexicana amazonensis. A simple and economical trapping technique for opossums is presented here, and a control method in special cases is suggested.Vinte e uma mucuras (Didelphis marsupialis), procedentes de uma floresta primária explorada, e sete de uma vegetação secundária em uma ilha isolada, nas cercanias de Manaus, foram examinadas para detectar a presença de Leishmania. Das mucuras da floresta primária, 13 (61.9%) foram detectadas positivas para Leishmania braziliensis guyanensis. Somente uma mucura foi dada como positiva para Le.mexicana amozonensis. Apresentamos aqui uma técnica simples e economica para captura de mucuras e e sugerido um método de controle, em casos especiais.

A total of 52 opossums (six species) were examined for evidence of infection with Leishmania in three different areas of forest near Manaus, Amazonas State, Brazil. No infections were detected in 27 opossums from a region of relatively undisturbed forest, including specimens of Didelphis marsupialis (18); Metachirus nudicaudatus (four); Monodelphis brevicaudata (one); Marmosa cinerea (two); M. murina (one) and M. parvidens (one). Of 15 D. marsupialis captured from a biological reserve, much disturbed by man, three were infected with L. braziliensis guyanensis: isolations were made from the skin of two of the animals, and from the viscera of the third. The isolates were biologically and biochemically indistinguishable from one isolate of L. b. guyanensis made from man and two from the sandfly vector Lutzomyia umbratilis from the same area. Two of eight D. marsupialis and both of two M. cinerea from another area of virgin forest used for army manoeuvres were infected with Leishmania mexicana amazonensis: the parasite was in all four cases isolated from normal skin. Five of nine specimens of Proechimys guyannensis, from the vicinity of Manaus, were also infected with L. m. amazonensis. A further 13 mammals (eight species) were negative for Leishmania. The importance of opossums as a reservoir of L. b. guyanensis is discussed. Although they may play only a minor role in virgin forest which is undisturbed by man, opossums (D. marsupialis) may become a significant reservoir of infection where man's activities have eliminated the major reservoir--which has yet to be incriminated.

In a survey of 296 sylvatic animals captured from virgin forests in the north-eastern and south-western Amazon of Brazil, Histoplasma capsulatum was isolated, via the indirect hamster inoculation method, from the liver and spleen of four common opossums Didelphis marsupialis and two pacas Agouti paca. The infected animals did not show any clinical symptoms or histopathology. The known Amazonian mammalian species with natural histoplasmosis now total five, the previously reported species being the spiny rat Proechimys guyannensis, the two-toed sloth Choloepus didactylus and the nine-banded armadillo Dasypus novemcinctus.

Paracoccidioides brasiliensis foi encontrado, por inoculação de triturado de fígado e baço em hamsters, em 4 de 20 tatus (Dasypus novemcinctus) examinados na região de Tucuruí, Pará. Hamsters inoculados por via intradérmica e peritoneal com o parasito desenvolveram infecções generalizadas e morreram em 1½ a 13 meses. A diagnose do fungo foi confirmada por histopatologia e cultura. Não se observaram sinais macroscópios de doenças nos tatus. A distribuição geográfica de D. novemcinctus abrange a área endêmica de paracoccidioidomicose humana, sugerindo-se que o tatu tenha algum papel na ecologia do fungo. In spite of an extensive literature on paracoccidioidomycosis, hardly anything is known about the ecology of Paracoccidioides brasiliensis in nature. During 1983, 152 wild animals of 21 species "were examined in a survey designed to detect sylvatic hosts of Leishmania near Tucuruí, a region of tropical rainforest with acid soils, in the State of Pará, northern Brazil. Hamsters inoculated with saline suspensions of liver and spleen from 4 out of 20 Dasypus novemcinctus developed generalized systemic infections after 4 to 13 months, with abundant spherical parasitic structures up to 30 mm indiameter, visible in unstained tissue smears. Inoculation of this material into fresh hamsters, produced lethal infections in within 1½ to 5 months, with gross pathological changes in the viscera and abundant parasites characteristic of P. brasiliensis in stained histological sections. Material from infected tissue grew slowly in Sabouraud Dextrose Agar, forming light-coloured cerebriform colonies approximately 1,5 cm in diameter after 2 months at 22-26ºC. Culture material was inoculated intradermally, intraperitoneally and intratesticularly into hamsters, laboratory mice and guinea pigs. Generalized infections were detected after approximately 5 months in female hamsters that had been inoculated intradermally. The fungus was re-isolated in culture from the infected hamsters. Parasites were detected in histological sections of the liver and spleen of the original armadillos, but no gross signs of disease. were noted in these animals. D. novemcinctus is widely distributed in the Neotropical Region but is absent from certain regions, such as Chile and Patagonia, where paracoccidioidomycosis is unknown. The fossorial habits of this armadillo may be relevant in the light of previous suggestions that the saprophytic phase of P. brasiliensis inhabits a subterranean environment. It is suggested that D. novemcinctus may play a part in the ecology of P. brasiliensis in nature.

Of 151 Trypanosoma cruzi stocks from 117 different individual hosts collected in the states of Amazonas and Rondonia, 147 from 113 hosts were identified as zymodeme 1 (Z1). These included T. cruzi stocks from three marsupial species, two rodent species and three triatomine species although most were from the common opossum, Didelphis marsupialis. One T. cruzi stock from Rhodnius robustus was identified as Z1 with a Z3 PGM character, one from Sciurus sp. as Z3 and two from Monodelphis brevicaudata and Panstrongylus geniculatus as Z3 with a Z1 ASAT character. The ways in which stocks were isolated and grown up in vitro did not influence isozyme profile. These results support earlier evidence from Pará State that Z2 is absent from the Amazon basin and that the distribution of T. cruzi zymodemes in this region is quite different to that in endemic areas on the south of the continent.

Isolement chez 3 sujets dans la regin nord de Manans ou la leishmaniose cutanee due a L. g. est frequente