Morphological Description of the Immature Stages of Nyssomyia umbratilis (Ward & Frahia) (Diptera: Psychodidae: Phlebotominae), the Main Vector of Leishmania guyanensis Floch (Kinetoplastida: Trypanosomatidae) in the Brazilian Amazon Region.

Nyssomyia umbratilis (Ward & Frahia) is the main vector of Leishmania guyanensis in the Brazilian Amazon region, where it is widely distributed. Studies have hypothesized that this phlebotomine is part of a cryptic species complex, spatially delimited by the Amazonian river systems, and with different transmission potential of L. guyanensis. In the present study, the immature stages of N. umbratilis are described from laboratory-reared specimens originating from the states of Amazonas and Pará, Brazil. Based on scanning electron and optical microscopy examinations, fine morphological structures (eggshell, setae, and mouthparts of larvae and pupae) are presented. A new form of sandfly pupae sexing (non-invasive) is also provided. The correct identification of insect is extremely important for the epidemiology of certain diseases. However, only the morphological characters of adult can be insufficient to separate accurately the closely related species. It is expected that the present description may contribute to solve the taxonomic problem involving N. umbratilis.

Comparative biology of two populations of Lutzomyia umbratilis (Diptera: Psychodidae) of Central Amazonia, Brazil, under laboratory conditions.

Lutzomyia umbratilis is the main vector of cutaneous leishmaniasis due to Leishmania guyanensis in northern South America. It has been found naturally infected with this species of Leishmania only east of the Rio Negro and north of the Rio Amazonas. However, populations of this sand fly species are also present in areas south of the Amazon river system, which may act as a geographical barrier to the Leishmania guyanensis cycle. With the aim of looking for possible biological differences between populations of L. umbratilis from each side of this river system, their biology in the laboratory was investigated. Progenitors collected on tree bases in Manaus and Manacapuru (east and west, respectively, of the Rio Negro) were reared in the laboratory. Results from observations of the life cycle, fecundity, fertility, and adult longevity at 27 degrees C and 92% RH were analyzed by descriptive statistics and z, t, U, and chi2 tests. Although the Manaus and Manacapuru colonies showed a longer developmental time than most Lutzomyia species reared at similar temperatures, length of time of egg and 4th instar larva of the two populations differed significantly (p < 0.01). Females of the latter retained significantly (p < 0.001) less mature oocytes, and the general productivity (% adults from a known number of eggs) of the colony was significantly (p < 0.01) higher than that of the former. These results show that the L. umbratilis population of Manaus is more productive, and thus a better candidate for future mass-rearing attempts. The two populations differ in their life cycle, fecundity, fertility, adult longevity, and emergence. These differences may reflect some divergence of intrinsic biological features evolved as a result of their geographical isolation by the Rio Negro. It is expected that further investigations on morphometry, cuticular hydrocarbon, isoenzyme, molecular and chromossomal analyses, infection, and cross-mating experiments with these and other allopatric populations of both margins of the Amazon river system will help reveal whether or not L. umbratilis has genetically diverged into two or more reproductively isolated populations of vectors or non-vectors of Leishmania guyanensis.

Comparative biology of two populations of Lutzomyia umbratilis (Diptera: Psychodidae) of Central Amazonia, Brazil, under laboratory conditions

Lutzomyia umbratilis é o principal vetor de leishmaniose tegumentar causada por Leishmania guyanensis no norte da América do Sul. Essa espécie tem sido encontrada naturalmente infectada com Leishmania somente ao leste do Rio Negro e norte do Rio Amazonas. Porém, populaçäes dessa espécie de flebotomíneo também estão presentes em áreas do sul do sistema fluvial do Rio Amazonas, o qual pode atuar como uma barreira geográfica no ciclo da Leishmania guyanensis. Com o objetivo de procurar possíveis diferenças biológicas entre populaçäes de L. umbratilis de margens opostas desse sistema fluvial, a biologia de duas populaçäes diferentes foi estudada em laboratório. Progenitores coletados em Manaus e Manacapuru (leste e oeste, respectivamente, do Rio Negro) foram criados separadamente. Resultados de observaçäes do ciclo de vida, fecundidade, fertilidade e longevidade de adultos a 27§C e 92% UR (umidade relativa) foram analisados por estatística descritiva, e testes z, t, U e c2. Embora as colônias de Manaus e Manacapuru tenham apresentado desenvolvimento mais demorado que a maioria das espécies de Lutzomyia, a duração das fases de ovo e de larva de 4§ estágio nas duas populaçäes foi significativamente (p < 0,01) diferente. Fêmeas de Manaus retiveram significativamente (p < 0.001) menos óvulos maduros, e a produtividade geral (% de adultos a partir de um número conhecido de ovos) da colônia foi significativamente (p < 0,01) mais alta do que a de Manacapuru. Estes resultados apontam a população de L. umbratilis de Manaus como a melhor candidata a futuras tentativas de criação em massa em laboratório. As diferenças observadas nas duas populaçäes quanto ao ciclo de vida, fecundidade, fertilidade, longevidade e emergência de adultos podem ser resultantes do isolamento geográfico ocasionado pelos grandes rios.

Molecular evolution of the period gene in sandflies.

The molecular evolution of the clock gene period was studied in Phlebotomine sandflies (Diptera: Psychodidae). Comparison of the synonymous and nonsynonymous substitution rates between sandflies and Drosophila revealed a significantly higher evolutionary rate in the latter in three of the four regions analyzed. The differences in rate were higher in the sequences flanking the Thr-Gly repetitive domain, a region that has expanded in Drosophila but remained stable and short in sandflies, a result consistent with the coevolutionary scenario proposed for this region of the gene. An initial phylogenetic analysis including eight neotropical sandfly species and one from the Old World was also carried out. The results showed that only the subgenus Nyssomyia is well supported by distance (neighbor-joining) and maximum parsimony analysis. The grouping of the other species from the subgenus Lutzomyia and Migonei group shows very low bootstrap values and is not entirely consistent with classical morphological systematics of the genus Lutzomyia.

Molecular evolution of the cacophony IVS6 region in sandflies.

A number of insects produce acoustic signals during courtship. Genes involved in the control of the courtship song are particularly interesting from an evolutionary viewpoint because interspecific variation in this signal is potentially important as a reproductive isolation mechanism and, as a consequence, in the speciation process. The cacophony gene was identified by a mutation affecting the "lovesong" in Drosophila melanogaster. Phlebotomine sandflies (Diptera: Psychodidae) also produce acoustic stimuli during courtship and therefore cacophony can be used as an interesting molecular marker in evolutionary studies in these important disease vectors. In this paper we have studied the molecular evolution of the IVS6 region of cacophony in sandflies. We compared the level of divergence in the exon sequences encoding this conserved domain in Drosophila and Phlebotomines. We also analysed the high level of variation in an intron that is present in sandflies but that was lost in Drosophila during evolution. The available cacophony sequences were also used for a phylogenetic analysis of some species of the Neotropical genus Lutzomyia.