Nataly A. Souza

Infecção natural de Lutzomyia intermedia Lutz & Neiva, 1912, em área endêmica de leishmaniose tegumentar no Estado do Rio de Janeiro

In a focus of cutaneous leishmaniasis in Jacarepagua, Rio de Janeiro, one specimen of Lutzomyia intermedia was found naturally infected with Leishmania braziliensis.

Studies on sandfly fauna (Diptera: Psychodidae) in a foci of cutaneous leishmaniasis in Mesquita, Rio de Janeiro State, Brazil

In a study on vectors of cutaneous leishmaniasis in Mesquita, Rio de Janeiro State, Brazil, eleven species of sandflies were found: Lutzomyia longipalpis, L. fischeri, L. quinquefer, L. intermedia, L. whitmani, L. shannoni, L. cortelezzii, L. edwardsi, L. migonei, L. lanei and L. firmatoi. Lutzomyia intermedia was the predominant species. It was collected indoors but in much higher numbers outdoors, in the close vicinity of houses. In comparative captures it was found to feed more frequently on equines, and less frequently on man and dogs. Studies, using man and animals as baits, showed that blood-feeding started in the evening, and remained stable during the night, until early morning. June, August and October were the months of highest density.

Lutzomyia longipalpis in Brazil: a complex or a single species? A mini-review

Lutzomyia longipalpis is the main vector of Leishmania infantum chagasi, the causative agent of American visceral leishmaniasis (AVL). Although there is strong evidence that Lu. longipalpis is a species complex, not all data concerning populations from Brazil support this hypothesis. The issue is still somewhat controversial for this large part of Lu. longipalpis distribution range even though that it is the Latin American region contributing to most of the cases of AVL. In this mini-review we consider in detail the current data for the Brazilian populations and conclude that Lu. longipalpis is a complex of incipient vector species with a complexity similar to Anopheles gambiae s.s. in Africa.

Seasonality of Lutzomyia intermedia and Lutzomyia whitmani (Diptera: Psychodidae: Phlebotominae), occurring sympatrically in area of cutaneous leishmaniasis in the State of Rio de Janeiro, Brazil

Lutzomyia intermedia and Lutzomyia whitmani occur sympatrically in the district of Posse, municipality of Petrópolis, in the State of Rio de Janeiro (114 km from the city of Rio de Janeiro), where sand flies were captured weekly for three consecutive years in both the peridomicile and forest, in two first years from 18:00 to 22:00 h, using a Shannon trap and CDC light traps. The anthropophilic habit was assessed by recording the specimens that bit the human captors. Considering the different capture types and sites, the following species were identified: Brumptomyia brumpti, B. cardosoi, B. guimaraesi, Lutzomyia amarali, L. longipalpis, L. microps, L. costalimai, L. edwardsi, L. firmatoi, L. migonei, L. termitophila, L. tupynambai, L. fischeri, L. pessoai, L. lutziana, L. shannoni, L. barrettoi, L. intermedia, L. whitmani, L. lloydi, and L. minasensis. L. intermedia (38.3%) and L. whitmani (38.6%) predominated. L. intermedia predominated in the peridomicile, where it displayed more intense anthropophily, while L. whitmani displayed greater density in the banana plantations and forest, preferential sites for biting humans. Studies on seasonality showed that while L. intermedia displayed greater density in the hottest months of the year (December, January, and February), L. whitmani was abundant in the coolest months (June, July, and August), although both occurred throughout the year.

The period gene and genetic differentiation between three Brazilian populations of Lutzomyia longipalpis.

Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae), the main vector of visceral leishmaniasis in the Americas, is a putative species complex. Molecular polymorphism was characterized in a 266 bp fragment of L. longipalpis homologous to period, a ‘speciation gene’ from Drosophila. Samples from the Brazilian localities of Jacobina (BA), Lapinha (MG) and Natal (RN) were analysed and the data indicate that the three populations are highly differentiated, with a very low level of gene flow between them. These results are in agreement with published pheromone and copulation song studies that suggest the existence of a sibling species complex in Brazil.

Genetic identification of two sibling species of Lutzomyia longipalpis (Diptera: Psychodidae) that produce distinct male sex pheromones in Sobral, Ceará State, Brazil.

Lutzomyia longipalpis, the main sandfly vector for New World visceral leishmaniasis is a complex of an as yet undefined number of sibling species. At present, there is no consensus on the status (single species vs. species complex) of Brazilian populations. We applied five microsatellite loci to test the hypothesis that L. longipalpis occurs as two sympatric cryptic species in Sobral, Ceará State, Brazil as predicted by male sex pheromone chemotypes described previously for field specimens from this site [S‐9‐methyl‐germacrene‐B (9MGB) and a cembrene compound]. Abdominal spot morphology corresponds with pheromone type at this locality (9MGB in ‘1 spot’ males and cembrene in ‘2 spot’ males). Genotype data from 190 wild‐caught L. longipalpis specimens collected in October 1999 and April 2001 were used to estimate genetic differentiation between the two sex pheromone populations and sampling dates. No significant (P > 0.05) genetic differences were found between the 1999 and 2001 9MGB samples (θ = 0.018; RST = −0.005), and genetic differentiation was low between the cembrene collections (θ = 0.037, P < 0.05; RST = −0.043, P > 0.05). By contrast, highly divergent allelic frequencies (largely at two microsatellite loci) corresponded to significant (P > 0.05) genetic differentiation (θ = 0.221; RST = 0.215) for all comparisons between samples with different pheromones. When pheromone samples were pooled across sample date, genetic differentiation was high (θ = 0.229; P < 0.001; Nem = 0.84). The allele frequency distribution at each of the five microsatellite loci was similar for males and females from the two collection years. Two of these loci showed highly divergent allele frequencies in the two sex pheromone populations. This was reflected in the highly significant genetic differentiation obtained from the male genotypes, between populations producing different pheromones (θ = 0.229–0.268; P < 0.0001 for the 2001 and θ = 0.254–0.558; P < 0.0001 for the 1999 collections, respectively). Similar results were obtained when the females, assigned to a pheromone type, were included in the analysis. Both a Bayesian analysis of the data set and a population assignment test provided strong evidence for two distinct populations corresponding to pheromone type. Given its genotype, the probability of assigning a 9MGB male to the original 9MGB population was 100% once the two years’ collections were pooled. For cembrene‐producing ‘2 spot’ males this probability although still high, was lower than for 9MGB males, at 86%. This microsatellite data together with previously reported reproductive isolation between the two Sobral populations confirm that premating barriers are important in speciation of L. longipalpis.

Molecular and behavioral differentiation among Brazilian populations of Lutzomyia longipalpis (Diptera: Psychodidae: Phlebotominae).

Background Lutzomyia longipalpis is the primary vector of American visceral leishmaniasis. There is strong evidence that L. longipalpis is a species complex, but until recently the existence of sibling species among Brazilian populations was considered a controversial issue. In addition, there is still no consensus regarding the number of species occurring in this complex. Methodology/Principal Findings Using period, a gene that controls circadian rhythms and affects interpulse interval periodicity of the male courtship songs in Drosophila melanogaster and close relatives, we analyzed the molecular polymorphism in a number of L. longipalpis samples from different regions in Brazil and compared the results with our previously published data using the same marker. We also studied the male copulation songs and pheromones from some of these populations. The results obtained so far suggest the existence of two main groups of populations in Brazil, one group representing a single species with males producing Burst-type copulation songs and cembrene-1 pheromones; and a second group that is more heterogeneous and probably represents a number of incipient species producing different combinations of Pulse-type songs and pheromones. Conclusions/Significance Our results reveal a high level of complexity in the divergence and gene-flow among Brazilian populations of the L. longipalpis species complex. This raises important questions concerning the epidemiological consequences of this incipient speciation process.

Analysis of the Copulatory Courtship Songs of Lutzomyia longipalpis in Six Populations from Brazil

Abstract The sand fly Lutzomyia longipalpis Lutz & Neiva (Diptera: Psychodidae: Phlebotominae), the main vector of Leishmania infantum in the Americas, is believed to be a species complex, although the status of different Brazilian populations is still somewhat unclear. Preliminary analysis of the acoustic signals that are produced during copulation by L. longipalpis males has suggested the existence of three sibling species in Brazil. In the current report, we analyze in more detail a number of parameters of the copulatory courtship songs of L. longipalpis males from four allopatric populations from different parts of the country (Marajó Island, Natal, Jacobina, and Lapinha Cave) and from two sympatric populations from the locality of Sobral, where two types of males can be differentiated by the number of pale spots (one or two pairs) found on the abdomen. We show that males from the localities of Natal, Marajó, and Sobral (two-spot morph) have very similar songs composed of successive bursts, which are modulated in frequency and amplitude. No significant differences were found in the song parameters of these three populations. In contrast, one-spot males from Sobral and males from Jacobina and Lapinha produce songs that are made of pulses but with distinct patterns for each population and significant differences in all song parameters studied. The results suggest that the L. longipalpis complex in Brazil is composed of four sibling species and that the differences in song patterns between the populations are consistent with the level of divergence found in the period gene.

Flebótomos de Vargem Grande, foco de leishmaniose tegumentar no Estado do Rio de Janeiro

In Vargem Grande, an endemic area of cutaneous leishmaniasis in State of Rio de Janeiro, where Lutzomyia intermedia had been found infected with Leishmania braziliensis, we performed a series of caputes of sandflies to increase the knowledge on their behaviour. The following species were found among 8,671 sandflies collected: L. intermedia, L. migonei, L. longipalpis, L. lanei, L. fischeri, L. firmatoi, L. monticola, L schreiberi, L. whitmani, L. pelloni, L. barrettpi e L. (Pi) sp. inside houses and outdoors close to the houses, on human bait and in light traps, the predominant species was L. intermedia, followed by L. migonei, both being more numerous between 9 and 11 p.m. than from 6 to 8 p.m. In a plantation, L. migonei was predominant. In simultaneous captures on man and dog as baits, L. intermedia was more frequent on the former, L. migonei on the latter.In Vargem Grande, an endemic area of cutaneous leishmaniasis in State of Rio de Janeiro, where Lutzomyia intermedia had been found infected with Leishmania braziliensis, we performed a series of captures of sandflies to increase the knowledge on their behaviour. The following species were found among 8,671 sandflies collected: L. intermedia, L. migonei, L. longipalpis, L. lanei, L. fischeri, L. firmatoi, L. monticola, L. schreiberi, L. whitmani, L. pelloni, L. barrettoi e L. (Pi) sp. Inside houses and outdoors close to the houses, on human bait and in light traps, the predominant species was L. intermedia, followed by L. migonei, both being more numerous between 9 and 11 p.m. than from 6 to 8 p.m. In a plantation, L. migonei was predominant. In simultaneous captures on man and dog as baits, L. intermedia was more frequent on the former, L. migonei on the latter.

The biological clock of an hematophagous insect: locomotor activity rhythms, circadian expression and downregulation after a blood meal.

Despite the importance of circadian rhythms in vector‐borne disease transmission, very little is known about its molecular control in hematophagous insect vectors. In Drosophila melanogaster, a negative feedback loop of gene expression has been shown to contribute to the clock mechanism. Here, we describe some features of the circadian clock of the sandfly Lutzomyia longipalpis, a vector of visceral leishmaniasis. Compared to D. melanogaster, sandfly period and timeless, two negative elements of the feedback loop, show similar peaks of mRNA abundance. On the other hand, the expression of Clock (a positive transcription factor) differs between the two species, raising the possibility that the different phases of Clock expression could be associated with the observed differences in circadian activity rhythms. In addition, we show a reduction in locomotor activity after a blood meal, which is correlated with downregulation of period and timeless expression levels. Our results suggest that the circadian pacemaker and its control over the activity rhythms in this hematophagous insect are modulated by blood intake.