AbstractBackground The reintroduction of sylvatic yellow fever in the state of São Paulo after about six decades was confirmed in the Northwestern region in 2000, where in 2008 there also occurred an important epizootic. The purpose of this study was to investigate the feeding habits of culicids potentially involved in the sylvatic transmission of the virus in this region.Methods Specimens were collected in 24 forested localities at ground level with hand nets and mouth aspirators. Collections were made quarterly between October 2006 and July 2008 during daylight hours. Blood-meal identification was carried out in mosquitoes of the tribes Aedini, Mansoniini and Sabethini. The biotin/avidin sandwich ELISA was employed to determine six source types: bird, bovine, equine, rat, human and monkey.Results A total of 24,879 females of the three tribes were obtained, 245 (0.98%) of which were engorged. The presence of three different blood sources per engorged female was the predominant situation, and included 35.10% of the total of samples processed. Samples with two or four different sources were represented by 25.31% and 25.71%, of the specimens, respectively, while just 9.39% had only one type and 1.22%, five different sources. Aedes scapularis, Ae. serratus(Group),Psorophora albigenu and Ps. ferox were the most abundant species and accounted for about 95% of the engorged specimens. Of the principal vector species,Haemagogus janthinomys/capricorniiwas found with bird, bovine and primate blood. These sources were predominant and alternated top ranking as the most frequent source according to the mosquito species and collection site. In general, primate blood was the most prevalent source.Conclusions The human population of the region visits this ecotone frequently, which indicates the need for the periodical assessment of vaccination coverage against yellow fever. The frequency of non-human primate blood source in mosquito species that show minor vector importance in yellow fever virus transmission deserves attention. The eclectic feeding habits and some aspects of the interactions between potential vectors and reservoirs of yellow fever may be associated with the habitat fragmentation characteristic of the region. We recommend that further studies on the capacity and vector competence be performed on secondary vectors in extra-Amazonian region.

Introduction The aim of this study was to identify the blood feeding sources of Nyssomyia intermedia (Ny. intermedia) and Nyssomyia neivai (Ny. neivai), which are Leishmania vectors and the predominant sandfly species in the Ribeira Valley, State of São Paulo, Brazil, an endemic area for cutaneous leishmaniasis. Methods Specimens were captured monthly between February 2001 and December 2003 on a smallholding and a small farm situated in the Serra district in the Iporanga municipality. The blood meals of 988 engorged females were tested using the avidin-biotin immunoenzymatic enzyme-linked immunosorbent assay (ELISA). Seven blood meal sources were investigated: human, dog, chicken, bovine, pig, horse and rat. Results The results showed that among the females that fed on one or more blood sources, the respective percentages for Ny. intermedia and Ny. neivai, respectively, were as follows: human (23% and 36.8%), pig (47.4% and 26.4%), chicken (25.7% and 36.8%) and dog (3.9% and 0%), and the differences in the blood sources between the two species were statistically significant (p = 0.043). Conclusions Both species had predominant reactivity for one or two blood sources, and few showed reactivity indicating three or four sources. Many different combinations were observed among the females that showed reactivity for more than one source, which indicated their opportunistic habits and eclecticism regarding anthropic environmental conditions.

A identificação da fauna flebotomínea e de alguns aspectos ecológicos de suas populações em áreas freqüentadas por turistas no PEI, situado em reserva de mata Atlântica, constituem-se nos objetivos deste estudo. As capturas foram mensais de janeiro/2001 a dezembro/2002, com armadilhas automáticas luminosas em 13 ecótopos, incluindo cavernas, matas e peridomicílio e aspiração em tocas de tatus. Sem intervalos regulares, foram realizadas capturas com armadilhas de Shannon na mata e ambientes antrópicos, aspiração em paredes de cavernas e entre raízes e folhedo e em membros da equipe quando picados por flebotomíneos. No total foram capturados 891 flebotomíneos pertencentes a 21 espécies. Com as armadilhas automáticas luminosas capturou-se 19 espécies e 600 espécimes, 215 em ambiente antrópico (2,24 insetos/armadilha) e 385 em ambiente natural (1,46 insetos/armadilha). Brumptomyia troglodytes foi a mais abundante, com o índice de abundância das espécies padronizado = 0,705. Pintomyia monticola predominou nas armadilhas de Shannon, mostrando-se antropofílica e com atividade hematofágica diurna e noturna. Psathyromyia pascalei predominou nas aspirações, com a maioria dos espécimes aspirados de tocas de tatu. Das 11 espécies capturadas em cavernas, embora algumas possam ser consideradas troglófilas, a maioria usa este ecótopo como local de abrigo. Nyssomyia intermedia, Nyssomyia neivai e Migonemyia migonei, implicados na transmissão da leishmaniose tegumentar na Região Sudeste do Brasil foram capturados, todavia em tão baixa densidade que sugere risco mínimo da doença no PEI.

The identification of the sandfly fauna and investigation of some ecological aspects of its populations in areas frequented by tourists of the PEI, an Atlantic forest reserve with many caves, were the objective of this study. Captures were undertaken monthly from January 2001 to December 2002, with automatic light traps installed in 13 ecotopes, including caves, forests, domiciliary and peridomiciliary environments, and by aspiration in armadillo burrows. Additionally, although not at regular intervals, Shannon traps were installed in forests and anthropic environments, aspirations were made on cave walls, among roots and fallen leaves, and some insects were captured while biting researchers. A total of 891 sandflies belonging to 21 species were captured. Six hundred specimens representing 19 species were captured with light traps, 215 in anthropic (2.24 insects/trap) and 385 in extra-domiciliary (1.46 insects/trap) environments. Brumptomyia troglodytes was the most abundant species (the Standardised Index of Species Abundance = 0.705). Pintomyia monticola predominated in the Shannon traps and showed anthropophilic and diurnal activity. Psathyromyia pascalei predominated in the aspirations; the largest number being in armadillo burrows. Eleven species were captured in caves; although some might be troglophiles, the majority used these ecotopes as resting places. Nyssomyia intermedia, Nyssomyia neivai and Migonemyia migonei, implicated in the transmission of cutaneous leishmaniasis in the Southeastern Brazilian region, were all found, though in such low densities as to suggest minimal risk of the disease in the PEI.

O Parque Estadual Turístico do Alto Ribeira, com cerca de 250 cavernas, situado em reserva de floresta Atlântica, é uma importante atração turística na região do Vale do Ribeira, onde a leishmaniose tegumentar americana (LTA) é endêmica. Com o objetivo de investigar as espécies incômodas ao homem e/ou implicadas na transmissão de Leishmania, identificou-se a fauna flebotomínea e alguns aspectos ecológicos de suas populações no núcleo da caverna Santana. Mensalmente, de janeiro/2001 a dezembro/2002, foram realizadas capturas com armadilhas automáticas luminosas em 11 ecótopos, incluindo cavernas, florestas, área de camping e ambientes domiciliares, e com armadilhas de Shannon em mata. No total capturou-se 2.449 flebotomíneos, representando 21 espécies. Nas capturas com armadilhas automáticas luminosas, Psathyromyia pascalei e Psychodopygus ayrozai foram as mais abundantes. Com as armadilhas de Shannon branca e preta instaladas simultaneamente foram capturados 107 espécimes, representando 13 espécies, na preta (12 espécies) e na branca (6 espécies). Fêmeas de Psychodopygus geniculatus predominaram na preta (43,75%), e Psathyromyia lanei e Ps. ayrozai, igualmente (32,4%), na branca. Nyssomyia intermedia e Nyssomyia neivai, ambas implicadas na transmissão da LTA na região, também foram capturadas, Ny. intermedia, a 6ª mais abundante, predominou na área de camping aberto. Baixas freqüências de flebotomíneos foram observadas nas cavernas, onde Evandromyia edwardsi predominou. Destaca-se a captura de Lutzomyia longipalpis, principal vetor do agente da leishmaniose visceral americana, sendo o registro mais meridional da mata Alântica.

The Parque Estadual do Alto Ribeira (PETAR) with about 250 caves, in an Atlantic forest reserve, is an important ecotourist attraction in the Ribeira Valley, an endemic area of American cutaneous leishmaniasis (ACL). With the purpose of investigating Leishmania vector species bothersome to humans the sandfly fauna was identified and some of its ecological aspects in the Santana nucleus, captures were undertaken monthly with automatic light traps in 11 ecotopes, including caves, forests, a camping site and domiciliary environments, and on black and white Shannon traps, from January/2001 to December/2002. A total of 2,449 sandflies representing 21 species were captured. The highest values of abundance obtained in the captures with automatic light traps were for Psathyromyia pascalei and Psychodopygus ayrozai. A total of 107 specimens representing 13 species were captured on black (12 species) and white (6 species) Shannon traps set simultaneously. Psychodopygus geniculatus females predominated on the black (43.75%), and Psathyromyia lanei and Ps. ayrozai equally (32.4%) on the white. Nyssomyia intermedia and Nyssomyia neivai, both implicated in the transmission of ACL in the Brazilian Southeastern region, were also captured. Ny. intermedia predominated in the open camping area. Low frequencies of phlebotomines were observed in the caves, where Evandromyia edwardsi predominated Lutzomyia longipalpis, the main vector of the American visceral leishmaniasis, was aslo present. This is its most southernly reported occurrence in the Atlantic forest.

Phlebotominae (Diptera, Psychodidae) na Província Espeleológica do Vale do Ribeira: 3. Área de pousadas para turistas que visitam o Parque Estadual do Alto Ribeira (PETAR), estado de São Paulo, Brasil. O estudo teve como objetivo caracterizar alguns aspectos ecológicos da fauna flebotomínea em área endêmica de leishmaniose tegumentar situada do Bairro Serra, município de Iporanga, onde se situam pousadas para turistas que visitam o PETAR. As capturas foram realizadas em uma chácara e em um sítio, mensalmente, entre janeiro de 2001 e dezembro de 2003, com armadilhas automáticas luminosas (AAL) em chiqueiro, galinheiro e varanda do domicílio de ambos os locais, e no sítio, também com armadilhas de Shannon branca/preta. Capturou-se com as AALs um total de 87.224 flebotomíneos, representando 19 espécies, mais dois híbridos de Nyssomyia intermedia (Lutz & Neiva, 1912) e Nyssomyia neivai (Pinto, 1926) e dois espécimes anômalos. Ny. intermedia apresentou o índice de abundância das espécies padronizado = 1,0 e Ny. neivai = 0,935. Na chácara, ambas predominaram no chiqueiro, com a média geométrica de Williams/captura para Ny. intermedia = 63,7 espécimes e para Ny. neivai = 29,2, e no sítio, no galinheiro, Ny. intermedia = 402,6 e Ny. neivai = 116,2. Capturou-se nas armadilhas de Shannon branca/preta 863 flebotomíneos: Ny. intermedia (75,4%) e Ny. neivai (24,3%), as fêmeas sendo significativamente predominantes na branca. As altas freqüências de Ny. intermedia e Ny. neivai, ambas implicadas na transmissão de agente da leishmaniose tegumentar, indicam risco de transmissão da doença na área.

Phlebotomines (Diptera, Psychodidae) in the Speleological Province of the Ribeira Valley: 3. Area of hostels for tourists who visit the Parque Estadual do Alto Ribeira (PETAR), state of São Paulo, Brazil. The study characterizes some ecological aspects of the phlebotomine fauna in an endemic area of cutaneous leishmaniasis (CL) situated in the Serra district, Iporanga municipality where the hostels for tourists visiting the PETAR are located. Captures were undertaken on a smallholding and a small farm situated near the hostels, monthly between January/2001 and December/2003 with automatic light traps (ALT) in pigsty, hen-house and veranda of a domicile at the two sites, and in peridomicile of the small farm also with black/white Shannon traps. With the ALT a total of 87,224 phlebotomines representing 19 species and also two hybrids of Nyssomyia intermedia (Lutz & Neiva) and Nyssomyia neivai (Pinto) and two anomalous specimens were captured. The standardized index species abundance was for Ny. intermedia = 1.0 and Ny. neivai = 0.935. The highest frequencies of the smallholding occurred in the pigsty, the Williams' mean/capture for Ny. intermedia being 63.7 specimens and for Ny. neivai 29.2, and on the small farm, in the hen-house, Ny. intermedia 402.6 and Ny. neivai 116.2. A total of 863 phlebotomines (Ny. intermedia: 75.4%; Ny. neivai: 24.3%) were captured with black/white Shannon traps; females of both species being predominant in the white trap. The high frequencies of Ny. intermedia and Ny. neivai, both implicated in CL transmission, indicate the areas presenting risk of the disease.

Lutzomyia longipalpis e Lutzomyia almerioi, espécies integrantes da fauna flebotomínea da Serra da Bodoquena, no Estado de Mato Grosso do Sul, têm sido objeto de estudo devido às suas elevadas abundâncias no Assentamento Guaicurus, foco de leishmaniose tegumentar humana e visceral canina. Em pesquisas que vem sendo realizadas neste acampamento para a identificação de vetores destas parasitoses, foram capturados no período de 2002 a 2004, com armadilhas automáticas luminosas, instaladas em ambiente peridoméstico (galinheiro), 83 exemplares ingurgitados de Lutzomyia longipalpis e Lutzomyia almerioi. O presente estudo teve como objetivo a investigação do hábito alimentar para ave das fêmeas de ambas as espécies de flebotomíneos, mediante o emprego da técnica imunoenzimática de captura,comparando-se a reatividade durante os anos de 2002 a 2004. Dentre 57 amostras de Lutzomyia longipalpis e 26 de Lutzomyia almerioi, foram encontradas 72% reagentes para ave em Lutzomyia longipalpis e 96% em Lutzomyia almerioi, o que justifica o estudo do hábito alimentar na região, como medida de prevenção e instituição de vigilância epidemiológica.

Lutzomyia longipalpis and Lutzomyia almerioi, phlebotomine species from the fauna of Serra da Bodoquena, in the State of Mato Grosso do Sul, Brazil, have been studied, particularly due to the fact of their abundance and occurrence, the Guaicurus settlement, focus of human tegumentary and canine visceral leishmaniasis. In researches that are being carried out in this settlement for identifying the vectors of these parasitosis, 83 engorged females belonging to the species Lutzomyia longipalpis and Lutzomyia almerioi were captured with automatic light traps from 2002 up to 2004 in the peridomiciliary environment of the Guaicurus settlement (hennery).The aim of this study was the investigation on bird feeding habit of females of both the phlebotomine species by the enzyme-linked immunosorbent assay technique, comparing the reactivity during the period from 2002 up to 2004. Of the 57 samples of Lutzomyia longipalpis and 26 of Lutzomyia almerioi that have been tested, 72% from Lutzomyia longipalpis and 96% from Lutzomyia almerioi were reactive, which justifies the feeding habit study in the region as a prevention measure and the institution of an epidemiological survey.

A identificação de sangue ingerido pelos insetos é um importante parâmetro para elucidar aspectos ligados à transmissão de zoonoses, dentre elas, as leishmanioses. Dos métodos empregados para esclarecer a atração de vetores por animais que possam atuar como reservatórios dessas parasitoses, destacam-se os imunológicos. O estudo teve como objetivo, padronizar a técnica imunoenzimática de captura e titular amostras de sangue ingerido em fêmeas de flebotomíneos ingurgitadas de Lutzomyia longipalpis criadas em laboratório e alimentadas experimentalmente em rato. Em vista da alta sensibilidade, favorecida pelo sistema avidina-biotina, foi possível a realização de pelo menos noventa testes, de cada uma das amostras em duplicata, e constatar a presença de sangue para todas as amostras com períodos de 12 e 24 horas pós-ingestão, observando-se diferença significativa entre os respectivos títulos.

Bloodmeals taken by insects constitute an important parameter for clarifying aspects of the transmission of zoonoses, including leishmaniases. Immunological assays can be used to investigate the attraction of vectors to animals, which may be hosts of these parasitoses. The objective of this study was to standardize a sandwich enzyme-linked immunosorbent assay and titer samples with different time periods of digestion, in laboratory-bred Lutzomyia longipalpis fed on rats. In the light of the high sensitivity that the biotin-avidin method permits, the technique provided at least ninety repeat tests for each sample and identified recent bloodmeals taken by these insects. Bloodmeals were detectable up to 12 and 24h after blood ingestion, and a significant difference between these titers was observed.

Foram imunizados 26 macacos-pregos (Cebus apella) adultos, através da via intramuscular, com uma dose de 1,0 ml da vacina anti-rábica Fuenzalida & Palacios, inativada, produzida a partir de cérebros de camundongos lactentes, de uso veterinário, empregada nas campanhas de prevenção da raiva animal de cães e gatos. Os animais pertenciam a três grupos experimentais, previamente imunizados com vacina anti-rábica Fuenzalida & Palacios submetidos a diferentes esquemas de vacinação, e permaneceram em cativeiro durante o período de junho de 1996 a junho de 1997. A revacinação foi realizada em todos os animais. As amostras de soros foram obtidas aos 0, 30, 180 e 365.dias, e armazenadas à temperatura de -20ºC, e a dosagem dos anticorpos realizada através do teste simplificado da inibição da fluorescência. Verificou-se após a revacinação 17/25 (68%) dos animais pertencentes aos grupos I, II e III, que se apresentavam com títulos inferiores ao limite indicativo de soroconversão (<0.5 UI/mL) responderam com produção de anticorpos neutralizantes, e, os outros cinco que se apresentavam com títulos acima desse limite, mantiveram-se nesse patamar. Em relação ao período da resposta imune humoral, não ocorreram diferenças significante entre os Grupos I, II e III (p>0,05). A vacina anti-rábica Fuenzalida & Palacios induziu a resposta imune nos macacos-pregos, após revacinação com produção de anticorpos neutralizantes, iguais ou superiores a 0,5 UI/ml, porém, de curta duração; não constituindo assim, imunógeno apropriado para ser utilizado na rotina de imunização destes animais de difícil lide, mantidos em cativeiro.

Twenty-six capuchin monkeys (Cebus apella) were intramuscularly immunized with 1.0 ml dose of a veterinary inactivated suckling mouse brain rabies vaccine (SMBV) employed in campaigns for rabies prevention in dogs and cats. The animals belonged to three experimental groups previously vaccinated with SMBV and different schemes, were kept in captivity from June, 1996 to June, 1997. All animals received a booster dose. Serum samples were obtained at the 0th, 30 th, 180th and 365 th days and kept stored at -20ºC. The antibodies dosage was carried out through the simplified inhibition fluorescent test. After a booster dose 17/25 (68%) of animals belonged to groups I, II and III, that had neutralizing antibodies above 0.5 IU/ml produced a humoral immune response equal or higher than 0.5 IU/ml, and another five animals that had neutralizing antibodies higher than 0.5 IU/ml kept in these levels. In relationship to longer humoral immune response there is no statistical difference between all groups, G-I x G-II, G-I x GIII, G-II x G-III (p>0,05). The SMBV induced humoral immune response in capuchin monkeys after a booster dose, producing neutralizing antibodies equal to or higher than 0.5 IU/ml; however, they were short-lasting, being therefore not appropriate as an immunogen to be used routinely in the immunization of these animals which are difficult both to be dealed with and to be kept in captivity.