Regional and interspecific variations in the morphology of insects of the Lutzomyia intermedia complex (Diptera, Psychodidae, Phlebotominae)]

INTRODUCTION: Lutzomyia intermedia s. s. and L. neivai, usually considered as belonging to just one species, constitute a complex of species. The measurements of several anatomical structures of specimens of both groups, from Brazil, Paraguay, Argentina and Bolivia were analysed. METHOD: Thirty-nine structures were measured in specimens of both sexes, using a graduated ocular, analysing comparisons by analyses of variance (ANOVA). RESULTS: A significant deviation in the proportions of females and males showing each palpal formula and the influence of the region of origin of the flies were verified. Labrum and maxilary palps were longer in females and Antennomere III was longer of males. Several significant differences in the measurements of wings were noted, most of them greater in females. The proportion of spermathecae with simple heads was significantly greater in L. neivai than in L. intermedia. Significant differences in the length of genital pump and filaments between the species were also noted. DISCUSSION: The variation in the palpal formulae in both sexes show the risk of the use of this formula for the association between specimens of the two sexes. The differences in the lengths of palps and labrum between the sexes could be related to female blood feeding. The possible biological significance of the sexual differential relation length/width of wings is commented on. The different proportions of females of the two species with each spermathecae head shape may help in identification. The differences in the lengths of genital pumps and filaments and the relation between them could help in the identification of males, which is still difficult.     

Genetic diversity and population structure in the Leishmania guyanensis vector Lutzomyia anduzei (Diptera,Psychodidae) from the Brazilian Amazon

Lutzomyia (Nyssomyia) anduzei has been recognized as a secondary vector of Leishmania guyanensis in the Brazilian Amazon region. Since L. anduzei is anthropophilic, co-exists in areas of high leishmaniasis transmission and has been found infected with L. guyanensis, the understanding of the vector population structure and of the process responsible for it is paramount to the vector management and control efforts. In this study we analyzed 74 and 67 sequences of the COI and Cytb loci, respectively, from mitochondrial DNA, aiming to estimate the intra-population genetic variability and population structure in six L. anduzei samples from the Brazilian Amazon region. For COI, we found 58 haplotypes, low to high (F_ST = 0.0310–0.4128) and significant (P = 0.0033) genetic structure, and reduced gene flow among populations. The haplotype network yielded many reticulations that likely resulted from hypervariability in the locus. For Cytb, we observed 27 haplotypes, low to moderate (F_ST = 0.0077–0.1954) and nonsignificant (P > 0.05) genetic structure for the majority of comparisons and extensive gene flow among populations, in line with the haplotypes network data. AMOVA analysis indicated that most of the variation occurred within populations (83.41%, 90.94%); nevertheless, there were significant differences (Φ_ST = 0.0906–0.1659; P = 0.00098; P = 0.00000) among them for both loci. The Mantel test showed that the genetic structure is not associated to an isolation-by-distance (IBD) model in either of both loci. These data suggest that L. anduzei is genetically very diverse. The genetic structure lacking IBD may be due to adaptation to local habitats and the low dispersal capacity of the sandflies, and both could lead to population fragmentation and geographic isolation. These findings have important implications for epidemiology, surveillance and vector control and may be a first step in understanding the evolutionary history of this species.     

Copulation songs in three siblings of Lutzomyia longipalpis (Diptera: Psychodidae)

We present the results of recording male courtship songs of the sandfly Lutzomyia longipalpis. The striking differences in the songs from 3 Brazilian populations of this sandfly with 3 distinct male pheromones support the 3 sibling species previously proposed based on this characteristic.     

Pathogen screening and bionomics of Lutzomyia apache (Diptera: Psychodidae) in Wyoming, USA

Phlebotomine sand flies are vectors of bacteria, parasites, and viruses. Lutzomyia apache, a North American sand fly, was incriminated as a vector of vesicular stomatitis viruses (VSV) due to overlapping ranges of the sand fly and recent outbreaks of VSV. We report on the discovery of 2 populations of L. apache in Wyoming from Albany and Fremont counties. We attempted to isolate VSV and phleboviruses from sand flies from Albany County and screened select flies by polymerase chain reaction for Bartonella and blood meals. We did not isolate viruses or detect DNA from vertebrate hosts or Bartonella. Flies were also tested for insect pathogens and other microbes. We detected a Rickettsia sp. in all flies that were examined and a parasitic protozoon, Ascogregarina sp., from the midgut of a larva. Eustigmaeus lirella, a stigmaeid mite, parasitized 2 field-caught females.     

Susceptibility of Phlebotomus papatasi (Diptera: Psychodidae) to DDT in some foci of cutaneous leishmaniasis in Iran.

The susceptibility of Phlebotomus papatasi to DDT was studied in field surveys at localities in different areas of Iran during 1985-88. In many parts of Iran houses had been treated with DDT for malaria control (1950-68). Tests were carried out in localities where the application of DDT had been discontinued since 1969. This investigation showed that P. papatasi from Isfahan is more tolerant to DDT than flies from other areas, probably a manifestation of DDT resistance.     

Diversity and species composition of sand flies (Diptera: Psychodidae) in a Venezuelan urban focus of cutaneous leishmaniasis

The present study examined the spatial and temporal abundance and diversity of phlebotomine sand flies in an area of Venezuela that is an ancient focus of leishmaniasis. The study was conducted in 6 stations in urban localities in Trujillo City, located in northwestern Venezuela (9 degrees 22' 24" N, 70 degrees 26' 08" W), which is located in a mountain range in the Andean ecoregion (altitude = 600-1,010 m). During 1995-99, entomological surveys were conducted after and before the rainy season. Shannon light traps were operated from 1800 to 2000 h in peridomestic site trap locations. Twelve species were captured, and Lutzomyia youngi, L. ovallesi, L. scorzai, L. gomezi, L. lichyi, and L. shannoni occurred at all localities in each year. The abundance of these species showed low variation over time but high variation between localities. The S?rensen similarity index, used to compare diversity between years within each locality, ranged from 0.60 at Carmona to 0.84 at La Hacienda. Sand fly communities exhibited annual variation in species richness and diversity. Variations were affected more by changes in species abundance than by changes in species composition. Lutzomyia ovallesi, L. lichyi, and L. scorzai had the highest coefficient of variation between years (63, 38, and 23%, respectively).     

Genetic structure and divergence in populations of Lutzomyia cruciata,a phlebotomine sand fly (Diptera: Psychodidae) vector of Leishmania mexicana in southeastern Mexico

The low dispersal capacity of sand flies could lead to population isolation due to geographic barriers, climate variation, or to population fragmentation associated with specific local habitats due to landscape modification. The phlebotomine sand fly Lutzomyia cruciata has a wide distribution throughout Mexico and is a vector of Leishmania mexicana in the southeast. The aim of this study was to evaluate the genetic diversity, structure, and divergence within and among populations of Lu. cruciata in the state of Chiapas, and to infer the intra-specific phylogeny using the 3′ end of the mitochondrial cytochrome b gene. We analyzed 62 sequences from four Lu. cruciata populations and found 26 haplotypes, high genetic differentiation and restricted gene flow among populations (Fst = 0.416, Nm = 0.701, p < 0.001). The highest diversity values were recorded in populations from Loma Bonita and Guadalupe Miramar. Three lineages (100% bootstrap and 7% overall divergence) were identified using a maximum likelihood phylogenetic analysis which showed high genetic divergence (17.2–22.7%). A minimum spanning haplotype network also supported separation into three lineages. Genetic structure and divergence within and among Lu. cruciata populations are hence affected by geographic heterogeneity and evolutionary background. Data obtained in the present study suggest that Lu. cruciata in the state of Chiapas consists of at least three lineages. Such findings may have implications for vector capacity and hence for vector control strategies.     

Population dynamics of Lutzomyia shannoni (Diptera: Psychodidae) at the Patuxent National Wildlife Research Refuge, Maryland

The seasonal abundance and temporal patterns of the adult sand fly (Lutzomyia shannoni Dyar) were examined at the Patuxent National Wildlife Research Refuge, MD, from August 3, 2005, to July 29, 2006. A total of 138 (53 males, 85 females) L. shannoni was collected from 4 dry ice-baited traps set at the same 4 locations throughout the study. The male:female ratio was 1:2.4. All 4 traps, separated by a maximum distance of approximately 1.6 km, operated simultaneously on the collection dates. The collection dates were spaced apart by near weekly intervals during the months of expected sand fly activity. No collections occurred in December-February. August was clearly the period of peak adult abundance as the numbers collected were significantly greater during this month than any other month of collection. Results indicate the existence of a unimodal pattern of abundance with adult emergence beginning in June and ending by September. The temporal pattern and abundance differ from what has been observed for the species on Ossabaw Island, a barrier island located along coastal Georgia, and at San Felasco Hammock Preserve State Park, Gainesville, FL. Continued research is needed to compile multiyear data to confirm the temporal abundance patterns of this species in Maryland.     

Blood feeding sources and peridomiciliation of Lutzomyia longipalpis (Lutz & Neiva, 1912) (Psychodidae, Phlebotominae)

A precipitin test was employed to study the alimentary tract content of Lutzomyia longipalpis in the intra- and peridomiciliary environments in the municipality of Raposa, Maranh o State, a transmission area for visceral leishmaniasis or kala azar. Out of 2,240 female sandflies captured, 547 (24.4%) had fed on vertebrate blood, with the following proportions: avian (87.9%); rodent (47.2%); human (42.4%); canine (27.6%); opossum (26.6%); and equine (22.5%). Based on a survey of 120 human dwellings, chickens were found to be the most common domestic animals in the peridomicile (28.3%), followed by dogs (21.7%), cats (17.5%), donkeys (13.3%), pigeons (7.5%), rabbits (3.3%), ducks (3.3%), and horses, mallards, and pigs (1.7% each). Synanthropic animals included opossums (39.3%), followed by rats (37.9%), bats (14.3%), raccoons (3.6%), foxes (2.1%), snakes (1.4%), and frogs (1.4%). The peridomiciliary presence of domestic and synanthropic animals as well as sandflies that had fed on human, opossum, and canid blood supports the hypothesis that kala azar transmission has been taking place in the anthropic environment in the municipality of Raposa.     

Digestive tract microbiota in female Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera: Psychodidae) from colonies feeding on blood meal and sucrose plus blood meal

There are very few reports on the microbiota of the digestive tract of sand flies, an important omission considering that blood is not the only meal ingested. Male and female sand flies obtain sugar meals from several sources, thereby increasing their chance of infection with microorganisms. Chances of contamination are higher when insects are bred in the laboratory, and this may affect the development of Leishmania spp. From the digestive tract of 300 sand fly females separated in two groups we isolated 10 species of bacteria in group 1 and 8 species in group 2. In group 1, Enterobacteriaceae of the following genera were identified: Serratia, Enterobacter, and Yokenella and the non-fermenters: Pseudomonas, Acinetobacter, and Stenotrophomonas. In group 2, the Enterobacteriaceae Enterobacter and Serratia were identified as well as the non-fermenters Acinetobacter, Stenotrophomonas, Burkolderia, and Pseudomonas.     

Assessment of sand fly (Diptera, Psychodidae) control using cypermethrin in an endemic area for visceral leishmaniasis, Montes Claros, Minas Gerais State, Brazil

Montes Claros in Minas Gerais State, Brazil, was considered an intense transmission area for visceral leishmaniasis. This study evaluated sand fly fauna after insecticide application. Captures were performed in 10 districts from September 2005 to August 2006 with CDC light traps inside and outside each residence. Cypermethrin was sprayed in two cycles during November/2005 and May/2006. The 636 specimens collected, belonging to 10 species, were predominantly Lutzomyia longipalpis (79%), and most frequently males (70%). The highest percentage of specimens were captured in areas surrounding domiciles (85.8%). The main species were observed to be sensitive to treatment with the insecticide. The results showed a reduction in the number of sand flies collected after use of cypermethrin in homes and annexes, and with residual effect lasting from two to four months.     

Leishmaniasis in Brazil. XXI. Visceral leishmaniasis in the Amazon Region and further observations on the role of Lutzomyia longipalpis (Lutz & Neiva, 1912) as the vector

Further evidence is presented incriminating the sandfly Lutzomyia longipalpis as the vector of Leishmania chagasi, the causative agent of American visceral leishmaniasis, in the Amazon Region of Brazil. During an outbreak of the disease in Santarém, Pará State, this insect was shown to be the only species of sandfly consistently present in and around the patient's homes, where it often occurred in very large numbers. Of 491 specimens dissected, 35 (7.14%) proved to be infected, and isolates of L. chagasi were made from 16 of 27 of these sandflies following the inoculation of the promastigotes into hamsters. Finally, the parasite was transmitted to four other hamsters which had been subjected to the bites of large numbers of wild-caught Lu. longipalpis. Isolates of Leishmania from Lu. longipalpis captures in Santarém, and in another focus of visceral leishmaniasis on the Island of Marajó, Pará, have been shown to be biologically and biochemically indistinguishable from the parasite infecting man, dogs and foxes in Pará, and from stocks obtained from man elsewhere in Brazil (Bahia and Ceará States).     

American cutaneous leishmaniasis in the sugar cane-producing area of Pernambuco, North-East Brazil

The cases of American cutaneous leishmaniasis reported here were observed between 1995 and 1998 in outpatients at a health clinic in the sugar cane-producing region in the south of Pernambuco State (Brazil), in which the disease is known to be endemic. Two hundred thirty eight cases were reported, equivalent to about ten cases per thousand patients. The disease affected patients of all ages, with a slight male predominance. The lesions were generally single and located on the legs. They were successfully treated with Glucantime. Vectors of the disease, sandflies of the genus Lutzomyia, were captured in and around homes, particularly in stables. This field work demonstrates the extent of a long-standing endemic that can be effectively treated within motivated health structures.     

First detection of Leishmania major DNA in Sergentomyia (Sintonius) clydei (Sinton, 1928, Psychodidae: Phlebotominae), from an outbreak area of cutaneous leishmaniasis in Tunisia

In recent years there has been growing interest in Sergentomyia species. Their role in the spread of mammalian leishmaniasis appears repeatedly in the literature and the possibility of its implication in Leishmania transmission to humans remains controversial. Sergentomyia (Sintonius) clydei is one of several cryptic species sharing therefore common morphologic criteria with others species of the subgenera Sintonius. Little is known about this specie in Tunisia.We sampled and identified different specimens including four specimens of S. clydei collected from Sidi Bouzid and Kairouan areas (center of Tunisia) using morphological tools. Male Sergentomyia clydei and Sergentomyia christophersi are known to share several morphological characters and can be mistaken for. Consequently we took advantage of 5 male S. christophersi available in our collection (Tataouin, South of Tunisia). In our study morphological tools were completed by molecular study of cytochrome b gene to identify S. clydei.For the detection of Leishmania spp. that might infect our specimens, Leishmania DNA was analyzed by amplification of kinetoplast minicircle DNA using real-time PCR and nested-PCR. Obtained result was confirmed by restriction analysis of the amplified ribosomal internal transcribed spacer 1 (ITS1).We provide in our study, the first molecular identification of S. clydei, in Tunisia. Our Neighbor Joining tree based on mitochondrial cytochrome b gene shows two different clusters. The first includes the Tunisians S. clydei and other specimens from Africa, Middle East and the Arabic peninsula, and the second cluster containing the specimens from Seychelle.Unexpectedly, we also demonstrate the infection of one anthropophilic female S. clydei by Leishmania major DNA. This finding shows that more attention should be paid when identifying parasites by molecular tools within sandfly vector.