Alguns aspectos citológicos da Leptospira Icterohaemorrhagiae em microscópio eletrônico

Os autores estudando culturas de 11 e 12 meses de Leptospira icterohaemorrhagiae, mantidas em meio de Schüffner modificado por Verwoort, observaram em microscópio eletrônico, corpos de leptospira com zonas de condensação do citoplasma assumindo aspecto granular Figs. 1, 3, 4, 9, 7, 12. Nas Figs. 6 e 7 vemos com nitidez um dêsses grânulos desmembrando-se do corpo da leptospira e cuja imagem é semelhante a outros grânulos esparsos. O número dessas formações é variável, parecendo depender de muitas causas. A deformação do corpo da leptospira parece depender da perda de consistência da membrana em virtude de alteração da sua permeabilidade, fenomenos que deve preceder á formação dos grânulos. Notamos também nas mesmas condições de experiência, como na Fig. 3, que os referidos grânulos tem dois aspectos diferentes: um é o que já referimos de forma irregular e opaco como se fôsse uma imagem de cromatina condensada e outro aspecto é também de forma irregular, porém, transparente como se houvesse esvaziado seu conteúdo primitivo, Figs. 5, 7 e 13. Na fig. 12, observamos o esvaziamento de uma forma destas o retraimento do seu conteúdo em outra, observação facilitada com auxílio de lente. A fig. 2 revela um aspecto frqüente encontrado em trabalhos desta natureza como no de CZEKALWSKI, J. W. e EAVES, G., isto é: formação de um grânulo com posterior segmentação do mesmo. Finalmente a Fig. 11 nos mostra grânulos com a mesma densidade que os fragmentos dos corpos da leptospira. Como dissemos acima, a cultura contendo material com êste aspecto foi inoculada em cobaias cuja hemocultura nos forneceu de novo o germe reisolado, o que faz admitir constituirem êsses grânulos uma fase no ciclo vital da leptospira ou então elemento capazer de manter condições e fatôres necessários à continuidade da espécie.

Bases experimentales para la identificacion de Leishmania spp. de America por morfometria de amastigotos

Estudios morfométricos sobre los amastigotos de dieciocho poblaciones de cuatro aislados pertenecientes a dos especies venezolanas de Leishmania (L. braziliensis y L. garnhami) indican que la posición del einetoplasto nose modifica en modo estadísticamente significativo cuando los parásitos son sometidos a pasajes por hamsteres y ratones,cultivos en medio NNN o por infección en un vector (Lu. townsendi). La posición del cinetoplasto, medido como la distancia entre el extremo posterior del amastigoto al cinetoplasto, dividido entre la distancia del organoide al extremo anterior de la célula, permite diferenciar a L. braziliensis de L. mexicana y L. garnhami. Los otros parámetros morfométricos no son tan confiables.

Factores dermicos que condicionan la infeccion de Lutzomyia townsendi (Ortiz, 1959) por Leishmania spp. de Venezuela

Se estudia la susceptibilidad de Lutzomyia townsendi a la infección con Leishmania spp. sobre lesiones experimentales de hamsteres. Se estudia la frecuencia y distribución de los amastigotos en la dermis, relacionándola con la profundidad que alcanza el estilete bucal del insecto. Una correlación positiva, con significante coeficiente de correlación, se establece entre la frecuencia de los parásitos a una profundidad de 100-150 nm en la dermis y el éxito de la infección de los flebótomos.

Ensayos metodologicos para la investigacion de reservorios de Leishmania spp en los Andes venezolanos

Se describen dos técnicas, presuntiva y confirmativa, para la investigación de mamíferos que pudieran ser reservorios de Leishmania que parasitan al hombre. Se investigan los cambios en los títulos de inmovilización y aglutinación de promastigotos de cultivo por los sueros de animales normales y expuestos una o varias veces a la inoculación intradérmica de pequeñas dosis de promastigotos vivos. Se registra una caída de los títulos de aglutinación en los sueros de hamsteres, de Holochilus venezuelae y de Didelphis marsupialis después de la inoculación con L. mexicana mexicana de Panamá y de L. gamhami de la región de los Andes venezolanos. Se discute la natureza de estos fenómenos. Se han hecho xenodiagnósticos con Lutzomyia townsendi en Holochilus venezuelae y Sigmodon hispidus infectados experimentalmente com L. mexicana mexicana, L. mexicana amazonensis, L. braziliensis y L. garnhami. Las pruebas fueron leidas mediante el examen microscópico de las gotitas de heces excretadas entre las 108 y 132 horas después de la ingesta infectante, tras colorearlas con Giemsa. Se obtuvieron resultados positivos en 23% de los experimentos usando mamíferos con lesiones localizadas, dejando a los flebótomos ingurgitarse libremente sobre animales anestesiados que poseian una hasta varias lesiones localizadas.

Redescription of Nomimoscolex piraeeba Woodland, 1934 (Cestoda, Proteocephalidea), from the Amazon catfishes, Brachyplatystoma spp. with proposal of synonyms and invalidation of Endorchiinae and Endorchis

The proteocephalid species Nomimoscolex piraeeba Woodland, 1934, N. dorad (Woodland, 1935) and Endorchis piraeeba Woodland, 1934, from Brachyplatystoma spp., South American silurid fishes, are critically revised. It is concluded that they concern to one species, N. piraeeba. The Endorchiinae, a subfamily of Monticelliidae, and genus Endorchis are invalidated herein. The valid species of Endorchiinae, belonging to genus Muzophorus, M. admonticellia Woodland, 1934, M. pirarara Woodland, 1934 and M. woodlandi Rego, 1984, are transferred provisionally to Zygobothriinae.

A new species of Probursata Bravo-Hollis, 1984 (Mogenea: Heteraxinidae: Heteraxininae) parasite of Oligoplites spp. (Osteichthyes: Carangidae) from the coast of the state of Rio de Janeiro, Brazil

Probursata brasiliensis n. sp., a gill filament parasite of carangid fishes, O. palometa (Cuvier), Oligoplites saurus (Bloch & Schneider), and O. saliens (Bloch), from the Brazilian coast, is described and illustrated. The new species differs from Probursata veraecrucis Bravo-Hollis, 1984, the type and only species of this genus by the presence of spines in the auricular expansions of the genital atrium, by the trifurcate supplementary process of the clamp's midsclerite, and by having a larger number of tests and clamps. This is the first record of the genus Probursata Bravo-Hollis, 1984, in the South Atlantic Ocean.

Monogeneans of leatherjackets, Oligoplites spp. (Osteichthyes: Carangidae), with the description of a new species of Metacamopia (Monogenea: Allodiscocotylidae) from the coast of the State of Rio de Janeiro, Brazil

Metacamopia oligoplites n. sp., a gill filament parasite of carangid fishes of three species of Oligoplites Gill, O. palometa (Cuvier), O. saurus (Bloch & Schneider), and O. saliens (Bloch), from the coast of the State of Rio de Janeiro, Brazil, is described and illustrated. Metacamopia oligoplites n. sp. differs from M. indica by: the shape of the body; the pre-, para-, and post-germarial testes; vaginas lacking sclerotized structures; well-developed seminal receptacles; muscular sleeves around the constriction between the vaginas and the seminal receptacles; and the haptor highly asymmetric, with a large, heel-like area; and differs from M. chorinemi by: the esophagus lacking diverticles; a larger number of testes (26-55) and not just, approximately 10; and the vaginas lacking sclerotized structures of any kind. This is the first record of Metacamopia in the South Atlantic Ocean. The generic diagnosis of Metacamopia is emended. Hargicola oligoplites is reported for the first time in the South Atlantic Ocean. Oligoplites palometa and O. saliens are new host records for Hargicola oligoplites.

Sensitivity of Leishmania spp. to Glibenclamide and 4-Aminopiridine: A Tool for the Study of Drug Resistance Development

We have demonstrated that Leishmania spp. grown as promastigotes, are sensitive to the K+ channel inhibitors 4-aminopyridine and glibenclamide. Their host cells, the macrophages, are not affected by similar concentrations of the drugs. We have also initiated the molecular characterization of the mechanisms involved in the development of drug resistance to glibenclamide by the parasite. Therefore, we have selected experimentally and begun to characterize the Venezuelan Leishmania (Leishmania) strain, NR resistant to glibenclamide [NR(Gr)]. The analysis of genomic DNA evidenced the existence of a fragment which apparently is amplified in NR(Gr). The fragment recognized by the pgpA probe, related to the Leishmania P-glycoprotein family and which was originally isolated from L. tarentolae, showed a size polymorfism between the sensitive and the resistant strain. These results suggest that the development of resistance to glibenclamide in the strain NR(Gr) might be associated with the amplification of the ltpgpA or related gene(s)

Use of Monoclonal Antibodies for the Identification of Leishmania spp. Isolated from Humans and Wild Rodents in the State of Campeche, Mexico

The genus Leishmania includes 30 described species which infect a wide variety of mammalian hosts. The precise identification of leishmanial parasites at the species level is very important in order to determine whether an organism, causing the disease in a given area, is of the same biotype as that found in suspected mammalian reservoirs. The objectives of the present study were (1) to identify leishmanial parasites isolated from humans and wild rodents from the State of Campeche, an endemic focus of localized cutaneous leishmaniasis (LCL) in southern Mexico, using an indirect immunofluorescent assay (IFA) with monoclonal antibodies (Mabs); and (2) to determine if the parasites of the two types of hosts were of the same biotype. All the wild rodents (six Ototylomys phyllotis, eight Oryzomys melanotis, five Peromyscus yucatanicus and two Sigmodon hispidus) and 96% (24/25) of the human isolates were identified as Leishmania (L.) mexicana confirming that this specific LCL focus is a wild zoonosis. The presence of one human isolate of L. (Viannia) braziliensis in the State of Campeche, confirmed the importance of an accurate taxonomic identification at species level.

A comparison of three DNA extractive procedures with Leptospira for polymerase chain reaction analysis

Three DNA extraction methods were evaluated in this study: proteinase K followed by phenol-chloroform; a plant proteinase (E6870) followed by phenol-chloroform; and boiling of leptospires in 0.1 mM Tris, pH 7.0 for 10 min at 100°C, with no phenol treatment. Every strain treated with proteinase K or E6870 afforded positive polymerase chain reaction (PCR) reaction. On the other hand, from five strains extracted by the boiling method, three did not feature the 849 bp band characteristic in Leptospira. We also evaluated by RAPD-PCR, DNAs from serovars isolated with proteinase K and proteinase 6870 with primers B11/B12. Each of the DNA samples provided PCR profiles in agreement with previous data. Moreover, the results with E6870 showed less background non-specific amplification, suggesting that removal of nucleases was more efficient with E6870. The limit for detection by PCR using Lep13/Lep14 was determined to be 10(2) leptospira, using the silver stain procedure.

In vitro activity of Brazilian strains of the predatory fungi Arthrobotrys spp. on free-living nematodes and infective larvae of Haemonchus placei

In vitro tests were carried out to assess the activity of 26 Brazilian isolates of predatory fungi of the genus Arthrobotrys on a free-living nematode (Panagrellus sp.) and on infective larvae of Haemonchus placei, a parasitic gastrointestinal nematode of cattle. The results showed that the free-living nematode Panagrellus sp. was the most preyed upon, compared to H. placei, for all the fungal treatments. Also, variable predatory capacity was observed for different fungal isolates belonging to the same genus when applied to different nematode species.