Correlates of the peridomestic abundance of Lutzomyia longipalpis (Diptera: Psychodidae) in Amazonian Brazil

Abstract. Abundance of the sandfly Lutzomyia longipalpis , the vector of Leishmania chagasi , was surveyed using CDC light-suction traps in fifteen villages and 180 homesteads on Marajo island, Para State, Brazil. Flies were most abundant in cultivated areas, secondary growth and open woodland, away from savanna grassland and forest. Within homesteads, the abundance of flies in animal sheds was weakly associated with the number of hosts kept therein, and inversely related to the number elsewhere, both human and animal. Houses harboured more flies if dogs were present and if the house had: a roof of thatch rather than of tiles or tin; a thatched roof which had not been treated with insecticide; relatively more holes in walls and roof. The sex ratio of flies was more male-biased at higher densities. We weigh the importance of these variables as risk factors for Le.chagasi infection.     

Carbohydrate digestion in Lutzomyia longipalpis' larvae (Diptera - Psychodidae)

Lutzomyia longipalpis is the principal species of phlebotomine incriminated as vector of Leishmania infantum, the etiological agent of visceral leishmaniasis in the Americas. Despite its importance as vector, almost nothing related to the larval biology, especially about its digestive system has been published. The objective of the present study was to obtain an overview of carbohydrate digestion by the larvae. Taking in account that phlebotomine larvae live in the soil rich in decaying materials and microorganisms we searched principally for enzymes capable to hydrolyze carbohydrates present in this kind of substrate. The principal carbohydrases encountered in the midgut were partially characterized. One of them is a α-amylase present in the anterior midgut. It is probably involved with the digestion of glycogen, the reserve carbohydrate of fungi. Two other especially active enzymes were present in the posterior midgut, a membrane bound α-glucosidase and a membrane bound trehalase. The first, complete the digestion of glycogen and the other probably acts in the digestion of trehalose, a carbohydrate usually encountered in microorganisms undergoing hydric stress. In a screening done with the use of p-nitrophenyl-derived substrates other less active enzymes were also observed in the midgut. A general view of carbohydrate digestion in L. longipalpis was presented. Our results indicate that soil microorganisms appear to be the main source of nutrients for the larvae.     

Studies on populations of Lutzomyia longipalpis (Lutz & Neiva, 1912) (Diptera: Psychodidae: Phlebotominae) in Brazil

Studies were performed on five Brazilian populations of Lutzomyia longipalpis: Salvaterra (PA), S?o José do Ribamar (MA), Canindé (CE), Natal (RN) and Gruta da Lapinha, Lagoa Santa (MG). No morphological differences were observed that could distinguish between these populations. Homogeneity tests showed that the allopatric populations display a certain heterogeneity and that the sympatric populations, with different patterns of spots, are homogeneous. The Student-Newman-Keuls test, represented by Euler-Venn diagrams, showed a disjunction between the populations from the north/northeast and the one from Gruta da Lapinha. Genetic distances between the four populations (excluding the Canindé population) were within the range of intrapopulational differences. The Gruta da Lapinha population displayed a heterozygotic deficiency that could be a consequence of high levels of inbreeding due to cryptic habits of living in a small cave. These results do not favor the hypothesis of a L. longipalpis species complex in Brazil, and the species should be considered high polymorphic.     

Stomodaeal valve ultrastructure in the sandfly Lutzomyia longipalpis (Diptera: Psychodidae)

The stomodaeal valve and adjacent cardia region of the alimentary canal of the phlebotomine sandfly Lutzomyia longipalpis have been studied using electron microscopy. With the scanning electron microscope (SEM), numerous pit-like structures of different sizes were found in the surface of the valve of male and female flies. Under the transmission electron microscope (TEM), the cells adjacent to the pores were shown to have numerous mitochondria and to lack neural innervation. It is suggested that the pit-like structures are probably secretory. A typical basiconic sensilla was found on the inner side of the oesophagus at the junction with the stomodaeal valve, with at least one dendrite which extended to the tip, indicating a chemosensory role. It is likely that this type of sensilla may control the movement of the stomodaeal valve after contact with fluids entering the mid-gut and may be responsible for diverting the sugar meal to the crop by closure of the valve.     

Lutzomyia maruaga (Diptera: Psychodidae), a new bat-cave sand fly from Amazonas, Brazil

A new species of parthenogenetic, autogenic and apparently extremely endemic phlebotomine is described from a sandstone cave located in primary terra firme forest to the North of the city of Manaus. Specimens were collected in the aphotic zone of the Refúgio do Maruaga cave by light trap and reared from bat guano. The adult morphology suggests a closer relationship to some Old World Phlebotominae than to species of Lutzomyia Fran?a encountered in the surrounding rainforest, but it shares characteristics with the recently proposed Neotropical genera Edentomyia Galati, Deanemyia Galati and Oligodontomyia Galati.     

First report of Lutzomyia (Nyssomyia) umbratilis Ward & Frahia, 1977 outside of Amazonian Region, in Recife, State of Pernambuco, Brazil (Diptera: Psychodidae: Phlebotominae)

Lutzomyia umbratilis, a known vector of Leishmania guyanensis in the north of Amazon basin, has been exclusively found in the Amazonian region. Here we report for the first time the occurrence of this species in northeastern Brazil. The epidemiological importance of the occurrence of this species in the Atlantic Forest is commented.     

Larval Breeding Sites of Lutzomyia longipalpis (Diptera: Psychodidae) in Visceral Leishmaniasis Endemic Urban Areas in Southeastern Brazil

Background

The scarcity of information on the immature stages of sand flies and their preferred breeding sites has resulted in the focus of vectorial control on the adult stage using residual insecticide house-spraying. This strategy, along with the treatment of human cases and the euthanasia of infected dogs, has proven inefficient and visceral leishmaniasis continues to expand in Brazil. Identifying the breeding sites of sand flies is essential to the understanding of the vector''s population dynamic and could be used to develop novel control strategies.

Methodology/Principal finding

In the present study, an intensive search for the breeding sites of Lutzomyia longipalpis was conducted in urban and peri-urban areas of two municipalities, Promissão and Dracena, which are endemic for visceral leishmaniasis in São Paulo State, Brazil. During an exploratory period, a total of 962 soil emergence traps were used to investigate possible peridomiciliary breeding site microhabitats such as: leaf litter under tree, chicken sheds, other animal sheds and uncovered debris. A total of 160 sand flies were collected and 148 (92.5%) were L. longipalpis. In Promissão the proportion of chicken sheds positive was significantly higher than in leaf litter under trees. Chicken shed microhabitats presented the highest density of L. longipalpis in both municipalities: 17.29 and 5.71 individuals per square meter sampled in Promissão and Dracena respectively. A contagious spatial distribution pattern of L. longipalpis was identified in the emergence traps located in the chicken sheds.

Conclusion

The results indicate that chicken sheds are the preferential breeding site for L. longipalpis in the present study areas. Thus, control measures targeting the immature stages in chicken sheds could have a great effect on reducing the number of adult flies and consequently the transmission rate of Leishmania (Leishmania) infantum chagasi.     

Ingestion of saliva during carbohydrate feeding by Lutzomyia longipalpis (Diptera; Psychodidae)

The aim of this study was to obtain experimental evidence that phlebotomine saliva is actually ingested during the carbohydrate ingestion phase (before and after blood digestion). The ingestion of carbohydrate was simulated as it occurs in the field by offering the insects balls of cotton soaked in sucrose, sucrose crystals or orange juice cells. The results obtained here showed that ingestion occurred under each condition investigated, as indicated by the presence of apyrase, an enzyme used as a marker to detect saliva in the insect gut and/or carbohydrate sources. Saliva ingestion by phlebotomine during the carbohydrate ingestion phase is important to explain how it could promote starch digestion and to trigger Leishmania promastigotes to follow a differentiation pathway as proposed previously by some authors.     

Moonlight and blood-feeding behaviour of Lutzomyia intermedia and Lutzomyia whitmani (Diptera:Psychodidae:Phlebotominae), vectors of American cutaneous leishmaniasis in Brazil

Lutzomyia intermedia (Lutz & Neiva, 1912) and L. whitmani (Antunes & Coutinho, 1939) (Diptera: Psychodidae: Phlebotominae), two important vectors of American cutaneous leishmaniasis in Brazil, occur in sympatry in the locality of Posse county, Petropolis municipality, state of Rio de Janeiro, Brazil. We investigated the influence of the lunar cycle on the frequency of specimens of the two species caught while attempting to bite the collectors and in CDC light traps. Analysis of the numbers of sand flies captured in different lunar phases for two consecutive years in the peridomestic site and forest shows that there is a significant positive correlation between moonlight intensity and the numbers of L. intermedia and L. whitmani females collected while blood-feeding, whereas the opposite was observed for the CDC traps.     

Navigation of Lutzomyia longipalpis (Diptera: Psychodidae) under dusk or starlight conditions

The responses of male and female Lutzomyia longipalpis (Lutz & Neiva) to different wavelengths of light was tested by presenting the sandflies with two light sources simultaneously, a series of test wavelengths between 350-670 nm and a 400 nm control. To test whether L. longipalpis could discriminate between the test and control, three sets of experiments were carried out in which the test wavelengths were presented at higher, equivalent or lower intensity than the control. In all three experiments, ultra-violet (350 nm) and blue-green-yellow (490-546 nm) light was more attractive to L. longipalpis than the control wavelength. However, at low intensity, UV was less attractive, than equivalent or higher intensity UV light. At intensities equivalent to or higher than the control wavelength, ultra-violet light was more attractive than blue-green. Furthermore, at low intensity, green-yellow (546 nm) light was more attractive to males whereas blue-green (490 nm) was more attractive to females. Blue-violet (400 nm) and orange-red (600-670 nm) light were least attractive in all three sets of experiments. Response function experiments indicated that the responses were dependent on both intensity and wavelength and that therefore more than one photoreceptor must be involved in the response. The results indicated that L. longipalpis can discriminate between different wavelengths at different intensities and thus have true colour vision. It also suggests that L. longipalpis may be able to navigate at dusk or under moonlight or starlight conditions using light in the blue-green-yellow part of the spectrum. The difference in response of males and females to light in this region is interesting and may indicate the different ecology of the sexes at night. Overall, these results may have important implications for sandfly trap design.     

Quantitative morphological evidence for incipient species within Lutzomyia quinquefer (Diptera: Psychodidae)

Morphological variation among geographic populations of the New World sand fly Lutzomyia quinquefer (Diptera, Phlebotominae) was analyzed and patterns detected that are probably associated with species emergence. This was achieved by examining the relationships of size and shape components of morphological attributes, and their correlation with geographic parameters. Quantitative and qualitative morphological characters are described, showing in both sexes differences among local populations from four Departments of Bolivia. Four arguments are then developed to reject the hypothesis of environment as the unique source of morphological variation: (1) the persistence of differences after removing the allometric consequences of size variation, (2) the association of local metric properties with meristic and qualitative attributes, rather than with altitude, (3) the positive and significant correlation between metric and geographic distances, and (4) the absence of a significant correlation between altitude and general-size of the insects.     

Sex pheromone and period gene characterization of Lutzomyia longipalpis sensu lato (Lutz & Neiva) (Diptera: Psychodidae) from Posadas, Argentina

Lutzomyia longipalpis s.l. is the primary vector of Leishmania (L.) infantum in the New World. In this study, male Lutzomyia longipalpis specimens from Posadas, Argentina were characterized for two polymorphic markers: the male sex pheromone and the period (per) gene. The male sex pheromone was identified as (S)-9-methylgermacrene-B, the same compound produced by Lu. longipalpis from Paraguay and many populations from Brazil. The analysis of per gene sequences revealed that the population from Argentina is significantly differentiated from previously studied Brazilian populations. Marker studies could contribute to the understanding of the distribution and spread of urban American visceral leishmaniasis, thus aiding in the design of regional surveillance and control strategies.     

Ootaxonomic investigation of five Lutzomyia species (Diptera, Psychodidae) from Venezuela

The eggshell fine structure of five sand fly species from Venezuela belonging to the genus Lutzomyia (L. migonei, L. ovallesi, L. absonodonta, L. gomezi and L. panamensis) was examined by scanning electron microscopy. The chorionic sculpturing of L. migonei, L. ovallesi, L. absonodonta and L. gomezi was characterized by series of columns arranged in palisade to form sinuous ridges. In inter-ridge areas, the basal layer was covered with fibrous material. The outer chorion of L. panamensis had a pattern known as "mountain- or volcano-like". The morphology of the posterior pole and aeropyle had a common structure in the five species, with some species-specific characters. The eggshell features of the five species are compared with those of other phlebotomine sand flies.