RESUMO
A set of 38 Leishmania stocks from the Andean valleys of Peru was characterized by both Multilocus Enzyme Electrophoresis (MLEE) and Random Amplified Polymorphic DNA (RAPD). Data were analyzed in terms of taxonomy and evolutionary genetics. Synapomorphic MLEE and RAPD characters, clear-cut clustering, and strong agreement between the phylogenies inferred from either MLEE or RAPD supported the view that Leishmania (Viannia) peruviana and Leishmania (Viannia) braziliensis correspond to two closely related, but distinct monophyletic lines (clades) and can therefore be considered as "discrete typing units" (DTUs). The question whether the L. (V.) peruxviana DTU deserves species status is dependent upon the desirability of it, in terms of epidemiological and medical relevance. A previous Orthogonal Field Alternating Gel Electrophoresis (OFAGE) analysis of the same L. (V.) peruviana isolates was published by Dujardin et al. (1995b). The data from the different markers (i.e. MLEE, RAPD and OFAGE) were compared by population genetics analysis. RAPD and OFAGE provided divergent results, since RAPD showed a strong linkage disequilibrium whereas OFAGE revealed no apparent departure from panmictic expectation. MLEE showed no linkage disequilibrium. Nevertheless, contrary to OFAGE, this is most probably explainable by the limited variability revealed by this marker in L. (V.) peruviana (statistical type II error). RAPD data were consistent with the hypothesis that the present L. (V.) peruviana sample displays a basically clonal population structure with limited or no genetic exchange. Disagreement between RAPD and OFAGE can be explained either by accumulation of chromosomal rearrangements due to amplification/deletion of repeated sequences, or by pseudo-recombinational events.
Assuntos
Leishmania/classificação , Animais , Eletroforese/métodos , Enzimas/genética , Genótipo , Humanos , Leishmania/enzimologia , Leishmania/genética , Peru , Fenótipo , Filogenia , Psychodidae/parasitologia , Técnica de Amplificação ao Acaso de DNA PolimórficoRESUMO
In the course of an epidemiologic survey in Ecuador, the following collection of Leishmania stocks was isolated: 28 from patients with clinical signs of leishmaniasis, 2 from sloths, 1 from a dog, and 4 from sand flies. For genetic characterization of these stocks, multilocus enzyme electrophoresis (MLEE) and random amplified polymorphic DNA (RAPD) were used. Twenty six of the 35 stocks were identified as either Leishmania (V.) panamensis or L. (V.) guyanensis, 2 stocks were identified as L. (V.) braziliensis, the 2 stocks from sloths showed specific genotypes, and 5 stocks were characterized as hybrids between L. (V.) braziliensis and L. (V.) guyanensis. These data show that genetic diversity of Leishmania in Ecuador is high and that L. (V.) panamensis/guyanensis is the dominant group in this country. The genetic analysis questioned the distinctness between the two species L.(V.) panamensis and L. (V.) guyanensis, since MLEE and RAPD data did not indicate that L. (V.) panamensis and L. (V.) guyanensis correspond to distinct monophyletic lines. Population genetic analysis performed on the L. (V.) panamensis/guyanensis group favors the hypothesis of a basically clonal population structure.
Assuntos
Variação Genética/genética , Leishmania guyanensis/genética , Leishmaniose Mucocutânea/parasitologia , Animais , Cães , Equador , Eletroforese em Acetato de Celulose , Glucosefosfato Desidrogenase/química , Humanos , Isoenzimas/química , Isoenzimas/genética , Leishmania guyanensis/classificação , Leishmania guyanensis/enzimologia , Leishmaniose Mucocutânea/enzimologia , Filogenia , Psychodidae , Técnica de Amplificação ao Acaso de DNA Polimórfico , Bichos-PreguiçaRESUMO
The taxonomic attribution of four Leishmania stocks isolated from humans in Ecuador has been explored by both multilocus enzyme electrophoresis and random amplified polymorphic DNA. For three loci, MLEE results showed patterns suggesting a heterozygous state for a diploid organism, while the corresponding homozygous states are characteristic of the Leishmania panamensis/guyanensis complex and Leishmania braziliensis, respectively. Other enzyme loci showed characters attributable to either the L. panamensis/ guyanensis complex or L. braziliensis. RAPD profiles exhibited for several primers a combination of the Leishmania panamensis/ guyanensis complex and L. braziliensis characters. These data hence suggest that the four stocks are the result of hybridization between L. panamensis/guyanensis and L. braziliensis. MLEE data show that the results cannot be attributed to either mixture of stocks, or an F1 in the framework of a simple Mendelian inheritance.
Assuntos
Quimera/genética , Leishmania braziliensis/genética , Leishmania guyanensis/genética , Filogenia , Animais , DNA de Protozoário/análise , Equador , Eletroforese em Acetato de Celulose/métodos , Humanos , Isoenzimas/análise , Leishmania braziliensis/enzimologia , Leishmania guyanensis/enzimologia , Leishmaniose Cutânea/parasitologia , Leishmaniose Mucocutânea/parasitologia , Técnica de Amplificação ao Acaso de DNA PolimórficoRESUMO
Lutzomyia trapidoi, the more abundant anthropophilic species, is a presumed leishmaniasis vector in the Pacific foothills of Ecuador. Three biotopes have been sampled (dwelling, and nearby coffee crop and primary forest) in the focus of Paraiso Escondido, by human bait catches, from August 1991 to October 1992. A large number of sandflies, 6,965 specimens, have been dissected to estimate peri and hypopyloric infections. All the peripyloric infections, characterized by isoenzyme electrophoresis, were Leishmania panamensis. The percentage of these infections was low, around 3%, but they were massive. They occurred only in dry season. Hypopyloric infections were observed in Lu. trapidoi all the year round in the three biotopes. Their percentage was high, reaching 40%. Despite of many trials to cultivate the parasite on NNN medium, no stain could be isolated. It is suggested that the parasite could be L. equatorensis.
Assuntos
Insetos Vetores/parasitologia , Leishmania guyanensis/isolamento & purificação , Psychodidae/parasitologia , Animais , Equador , Densidade Demográfica , Estações do AnoRESUMO
In two endemic leishmaniasis foci of the Pacific coast of Ecuador 34 dogs suspected of having the disease have been surveyed clinically, serologically and parasitologically; immunofluorescence and electrosyneresis tests, lymph node aspirates, biopsies and smears have been performed. From two dogs with ulcers only one had ulcers on the muzzle and the scrotum infected by Leishmania (L. guyanensis complex). The isolated strain was identified as Leishmania panamensis. The disease was strictly cutaneous. In the study area the dog seems to be more a victim-host than a reservoir.
Assuntos
Doenças do Cão/parasitologia , Cães/parasitologia , Leishmania guyanensis/isolamento & purificação , Leishmaniose Cutânea/veterinária , Animais , Equador , Leishmaniose Cutânea/parasitologiaRESUMO
Forty-one Leishmania peruviana isolates were selected along a north-south transect which crossed areas endemic for uta in three different biogeographical regions in the Peruvian Andes. The isolates were analysed by molecular karyotyping and hybridization with three chromosome-derived DNA probes. All the isolates could be distinguished from L. braziliensis by their pLb-134 hybridization patterns. However, the patterns with the other probes (pLb-168 and -22) could be used to cluster the Peruvian isolates in discrete groups (karyodemes) which varied in their level of similarity with L. braziliensis. The geographical distribution of these karyodemes supports the hypothesis that eco-graphical isolation has contributed to the heterogeneity of L. peruviana.