Emergence of phlebotomine sandflies (Diptera: Psychodidade) in non-flooded forest floor in Central Amazon, Brazil: a modified emergence trap model

Information concerning the potential natural breeding sites of phlebotomine sandflies are of high epidemiological importance. However, few studies have been conducted on the subject. This is due especially to the difficulties in finding as well as extracting immature sandflies that develop in the soil and organic matter of the forest floor. In the present study, a modified emergence trap model was tested in order to find potential breeding sites. This model was tested in the Pitinga Village, situated in the Presidente Figueredo municipality, in the State of Amazonas. Twenty-seven individuals belonging to nine species (Lutzomyia umbratilis,L. monstruosa,L. ayrozai,L. anduzei,L. trichopyga,L. davisi,L. geniculata,L. georgii e L. saulensis.) were collected. Lutzomyia umbratilis showed the highest number of individuals (37.1%) of all species captured in the area. The phlebotomine productivity was estimated as 2.2 sandflies per 100 m²/day. September showed the highest density of individuals, with a productivity of 5.8.Informações acerca de potenciais criadouros naturais de flebotomíneos sempre foram de fundamental interesse epidemiológico. Contudo, são poucas as informações advindas dos diversos estudos realizados até o momento. Isto se deve principalmente às dificuldades de localização e extração dos imaturos que se desenvolvem no solo e matéria orgânica do chão de florestas. No presente estudo o modelo modificado de armadilha de emergência foi testado na Vila do Pitinga, município de Presidente Figueiredo, Estado do Amazonas, a fim de localizar potenciais criadouros naturais. Vinte e sete indivíduos de nove espécies (Lutzomyia umbratilis,L. monstruosa,L. ayrozai,L. anduzei,L. trichopyga,L. davisi,L. geniculata,L. georgii e L. saulensis) foram coletados. Lutzomyia umbratilis foi a espécie com maior número de indivíduos, 10, representando 37,1% do total. A produção de flebotomíneos foi estimada em 2,2 flebotomíneos por 100 m² por dia. Em setembro, mês com maior número de indivíduos, esta produção foi de 5,8

A panel of ten microsatellite loci for the Chagas disease vector Rhodnius prolixus (Hemiptera: Reduviidae).

Rhodnius prolixus is the main vector of Chagas disease in Venezuela, where it is found colonising rural housing consisting of unplastered adobe walls with palm and/or metal roofs. Vector control failure in Venezuela may be due to the invasion of houses by silvatic populations of R. prolixus found in palms. As part of a study to determine if domestic and silvatic populations of R. prolixus are isolated, thus clarifying the role of silvatic populations in maintaining house infestations, we constructed three partial genomic microsatellite libraries. A panel of ten dinucleotide polymorphic microsatellite markers was selected for genotyping. Allele numbers per locus ranged from three to twelve, with observed and expected heterozygosity ranging from 0.26 to 0.55 and 0.32 to 0.66. The microsatellite markers presented here will contribute to the control of Chagas disease in Venezuela and Colombia through the provision of population information that may allow the design of improved control strategies

Isolation of

Natural infection with Leishmania spp. in phlebotomine sandflies was searched for during a longitudinal study carried out from July 1997 to July 1998 in the village Catarnica, Municipality Independencia, Táchira State. This hamlet is an old endemic focus of cutaneous leishmaniasis in the Venezuelan Andean region, which lies close to the Colombian border at 1,300 m a.s.l., in an agricultural area mainly used for cultivating coffee. Phlebotomine sandflies were collected using Shannon traps placed in the peridomestic habitat from 19:00 to 21:00 hs. Males were stored in alcohol 70% while females were kept in Nunc vials with 10% DMSO and cryopreserved in liquid nitrogen for subsequent dissection and identification. The most abundant anthropophilic species was Lutzomyia spinicrassa with 3,032 males and 4,290 females (85.4%). Among 1,633 (38%) females of Lu. spinicrassa dissected, 26 (1.6%) were infected with promastigotes, while no natural infection was found in 209 females of other species. The flagellates were identified as Leishmania braziliensis braziliensis using PCR with species specific primers derived from nuclear DNA and hybridization using species specific probe labelled with digoxigenin. This parasite had been previously isolated from patients with cutaneous leishmaniasis from the same area. These results show Lu. spinicrassa as a new proven vector of Leishmania braziliensis in the Andean region of Venezuela

Isolation of Leishmania (Viannia) braziliensis from Lutzomyia spinicrassa (species group Verrucarum) Morales Osorno Mesa, Osorno & Hoyos 1969, in the Venezuelan Andean Region

Natural infection with Leishmania spp. in phlebotomine sandflies was searched for during a longitudinal study carried out from July 1997 to July 1998 in the village Catarnica, Municipality Independencia, Táchira State. This hamlet is an old endemic focus of cutaneous leishmaniasis in the Venezuelan Andean region, which lies close to the Colombian border at 1,300 m a.s.l., in an agricultural area mainly used for cultivating coffee. Phlebotomine sandflies were collected using Shannon traps placed in the peridomestic habitat from 19:00 to 21:00 hs. Males were stored in alcohol 70% while females were kept in Nunc vials with 10% DMSO and cryopreserved in liquid nitrogen for subsequent dissection and identification. The most abundant anthropophilic species was Lutzomyia spinicrassa with 3,032 males and 4,290 females (85.4%). Among 1,633 (38%) females of Lu. spinicrassa dissected, 26 (1.6%) were infected with promastigotes, while no natural infection was found in 209 females of other species. The flagellates were identified as Leishmania braziliensis braziliensis using PCR with species specific primers derived from nuclear DNA and hybridization using species specific probe labelled with digoxigenin. This parasite had been previously isolated from patients with cutaneous leishmaniasis from the same area. These results show Lu. spinicrassa as a new proven vector of Leishmania braziliensis in the Andean region of Venezuela

Activity of Lutzomyia pseudolongipalpis and L. longipalpis s.l. (Diptera: Psychodidae) in Venezuela

The nocturnal activity of the phlebotomine sandfly Lutzomyia pseudolongipalpis and two populations of L. longipalpis s.l. from different American visceral leishmaniasis foci in Venezuela was studied using collection bottle rotator traps. The activity of L. pseudolongipalpis from Lara State was continuous throughout the night, while that of L. longipalpis s.l. from El Layero, Guàrico State and from Santa Ana del Valle, Margarita Island, was greatest before 23:00 h. The activity of sandflies of both populations and sexes steadily decreased thereafter. These different patterns seem to correlate with genetic data that indicate the presence in Venezuela of at least two sibling species in the L. longipalpis complex. The advantages of the bottle rotator trap for this type of study are discussed

Activity of

The nocturnal activity of the phlebotomine sandfly Lutzomyia pseudolongipalpis and two populations of L. longipalpis s.l. from different American visceral leishmaniasis foci in Venezuela was studied using collection bottle rotator traps. The activity of L. pseudolongipalpis from Lara State was continuous throughout the night, while that of L. longipalpis s.l. from El Layero, Guàrico State and from Santa Ana del Valle, Margarita Island, was greatest before 23:00 h. The activity of sandflies of both populations and sexes steadily decreased thereafter. These different patterns seem to correlate with genetic data that indicate the presence in Venezuela of at least two sibling species in the L. longipalpis complex. The advantages of the bottle rotator trap for this type of study are discussed

The re-emergence of American visceral leishmaniasis in an old focus in Venezuela. II . Vectors and parasites

As part of an epidemiological study in an old focus of American Visceral Leishmaniasis (AVL) in Venezuela (Guayabita, Aragua State], a longitudinal entomological survey (January 1993-June 1994) was carried out. A total of 3,239 males and 6,043 females belonging to 11 phlebotomine sandfly species were collected. The two recognised vectors of AVL in the New World, Lutzomyia evansi and Lu. longipalpis were found to be sympatric. Lutzomyia evansi was the dominant species (86.4 %), almost ten fold times more abundant than Lu. longipalpis (10.6 %). The two species alternated seasonally: Lu evansi peaked at the end of the rainy season while Lu. longipalpis, almost virtually absent during such period, increased in the dry season. This species seems more greatly influenced by the temperature. Seven of 4,559 Lutzomyia evansi (0.15 %) and one of 353 Lu. longipalpis (0.28 %) were found positive for suprapyloric promastigotes. Using the polymerase chain reaction (PCR) with universal primers, all isolates were identified as Leishmania spp. Two cultures from Lu. evansi, IEVA/VE/93/UCNA-2 and IEVA/VE/93/UCNA-3, were established. k-DNA restriction analysis showed high homologies between these isolates and Leishmania chagasi. High hybridization signal with Í. chagasi specific kDNA confirmed these results. These findings suggest that Lu. evansi may play a role as vector of visceral leishmaniasis in this area. The identity of the parasite carried by Lu. longipalpis needs to be confirmed

The re-emergence of American visceral leishmaniasis in an old focus in Venezuela: present situation of human and canine infections

The report of a new autochthonous case of human American Visceral Leishmaniasis (AVL) in 1992 in the village of Guayabita, Aragua State, Venezuela (10° 16'N, 67°28'W; 500 m asl), led us to undertake an epidemiological study in this locality. A demographic survey was conducted using a structured questionnaire in which data was collected regarding sex, age, occupation, length of residence in the area and migratory history. A leishmanin skin test (LST) was applied and samples of venous blood for counter immunoelectrophoresis (CIEP) and immunofluorescent antibodies (IFAT) tests were drawn. The prevalence of positive LST was 11.4 %. The positivity was highest among males and increased with age. Young males seemed to be more exposed to infection than females. Since occupation per se did not seem to account for this association, it may be explained as a gender-associated behavior, leading to different degrees of exposure to sand flies. The canine population was also screened for leishmanial infection. One dog was shown to be parasitologically infected with Leishmania sp. Four out of 71 dogs (5.6 %) were positive for FG, CIEP, IFAT and Western Blot and 11 (15.5 %) were positive for CIEP. These results suggest that after almost 30 years of epidemiological silence, American visceral leishmaniasis has re-emerged in this focus

Value of a dipstick based on recombinant rK39 antigen for differential diagnosis of American visceral leishmaniasis from other sympatric endemic diseases in Venezuela

A laboratory trial using recombinant rK39 dipsticks for differential diagnosis of American visceral leishmaniasis (AVL) from other sympatric endemic diseases which share similar clinic features (Chagas disease, malaria, schistosomiasis and toxoplasmosis) was conducted in Venezuela. The 100 % specificity of the test previously obtained in other countries was confirmed. The use of this test at the primary health care level in Venezuela for a rapid diagnosis of active AVL cases, which may avoid deaths, is recommended