Sandfly fauna and behavior (Diptera: Psychodidae) in municipalities of the Mesoregion North Pioneer of Paraná, Brazil

Abstract The occurrence of cutaneous leishmaniasis (CL) in the municipalities of Jaboti, Japira, Pinhalão, and Tomazina in the North Pioneer of the State of Paraná, where this disease is endemic, prompted the investigation of sandfly fauna, their population changes throughout the year, and their behavior in and around human accommodation. In these municipalities, the collection of sandflies was conducted using Falcão traps from 7 pm to 6 am once a month. 32,994 sandflies, 18,442 males and 14,552 females, were collected and represented by the species Brumptomyia brumpti, Brumptomyia cunhai, Evandromyia correalimai, Evandromyia cortelezzii, Expapillata firmatoi, Micropygomyia ferreirana, Migonemyia migonei, Nyssomyia neivai, Nyssomyia whitmani, Pintomyia fischeri, Pintomyia monticola, Pintomyia pessoai, and Psathyromyia bigeniculata. Ny. neivai (SISA = 0.9808) predominated in Japira (46.9%), Pinhalão (53.1%), and Tomazina (38.9%), while Ny. whitmani (SISA = 0.9423) predominated in Jaboti (53.8%). Most sandflies were captured in domestic animal shelters, with a peak in September. The high number of sandflies collected in domestic animal shelters, species richness, and presence of species involved in the epidemiology of CL reveal the need for permanent entomological surveillance in the municipalities mentioned. The use of phlebotomine control measures in these municipalities should be considered in conjunction with the anthropogenic actions that have resulted in changes to sandfly fauna and behavior.

Genetic variability of populations of Nyssomyia neivai in the Northern State of Paraná, Brazil

ABSTRACT The genetic study of sandfly populations needs to be further explored given the importance of these insects for public health. Were sequenced the NDH4 mitochondrial gene from populations of Nyssomyia neivai from Doutor Camargo, Lobato, Japira, and Porto Rico, municipalities in the State of Paraná, Brazil, to understand the genetic structure and gene flow. Eighty specimens of Ny. Neivai were sequenced, 20 from each municipality, and 269 base pairs were obtained. A total of 27 haplotypes and 28 polymorphic sites were found, along with a haplotypic diversity of 0.80696 and a nucleotide diversity of 0.00567. Haplotype H5, with 33 specimens, was the most common among the four populations. Only haplotypes H5 and H7 were present in all four populations. The population from Doutor Camargo showed the highest genetic diversity, and only this population shared haplotypes with those from the other municipalities. The highest number of haplotypes was sheared with Lobato which also had the highest number of unique haplotypes. This probably occurred because of constant anthropic changes that happened in the environment during the first half of the twentieth century, mainly after 1998. There was no significant correlation between genetic and geographical distances regarding these populations. However, the highest genetic and geographical distances, and the lowest gene flow were observed between Japira and Porto Rico. Geographical distance is a possible barrier between these municipalities through the blocking of haplotype sharing.

Disseminated cutaneous leishmaniasis caused by Leishmania braziliensis in Southern Brazil

ABSTRACT The authors report a case of disseminated cutaneous leishmaniasis caused by Leishmania (Viannia) braziliensis, in a 55 years old patient with 1,119 lesions distributed throughout the body. The patient resides in Sabáudia municipality, North of Paraná State, Southern Brazil, where there was no previous report of this form of leishmaniasis. Treatment with meglumine antimoniate was successful, although the diagnosis was made only five months later.

DETECTION OF Leishmania (Viannia) IN Nyssomyia neivai AND Nyssomyia whitmani BY MULTIPLEX POLYMERASE CHAIN REACTION, IN SOUTHERN BRAZIL / Detecção de Leishmania (Viannia) em Nyssomyia neivai e Nyssomyia whitmani por Multiplex Reação em Cadeia da Polimerase, no sul do Brasil

Sandflies transmit pathogens of leishmaniasis. The natural infection of sandflies by Leishmania (Viannia) was assessed in municipalities, in the state of Paraná, in Southern Brazil. Sandflies were collected with Falcão and Shannon traps. After dissection in search of flagellates in digestive tubes and identification of the species, female sandflies were submitted to the Multiplex Polymerase Chain Reaction (multiplex PCR) for detection of the fragment of the kDNA of Leishmania (Viannia) and the fragment from the IVS6 cacophony gene region of the phlebotomine insects. The analysis was performed in pools containing seven to 12 guts from females of the same species. A total of 510 female sandflies were analyzed, including nine Migonemyia migonei, 17 Pintomyia fischeri, 216 Nyssomyia neivai, and 268 Nyssomyia whitmani. Although none of the females was found naturally infected by flagellates through dissection, the fragment of DNA from Leishmania (Viannia) was shown by multiplex PCR in one sample of Ny. neivai (0.46%) and three samples of Ny. whitmani (1.12%). It was concluded that Ny. neivai and Ny. whitmani are susceptible to Leishmania infection, and that multiplex PCR can be used in epidemiological studies to detect the natural infection of the sandfly vector, because of its sensitivity, specificity and feasibility.

Flebotomíneos transmitem os patógenos das leishmanioses. Foi avaliada a infecção natural de flebotomíneos por Leishmania (Viannia) em municípios do Estado do Paraná, sul do Brasil. Os flebotomíneos foram coletados com armadilhas de Falcão e Shannon. Após dissecação para pesquisa de flagelados no tubo digestório e identificação das espécies, as fêmeas de flebotomíneos foram submetidas a Multiplex Reação em Cadeia da Polimerase (multiplex PCR) para a detecção do fragmento do kDNA de Leishmania (Viannia) e do fragmento do gene IVS6 da cacofonia de flebotomíneos. A análise foi realizada em pools contendo sete a 12 tubos digestórios de fêmeas da mesma espécie. Um total de 510 fêmeas foram analisadas, incluindo nove Migonemyia migonei, 17 Pintomyia fischeri, 216 Nyssomyia neivai e 268 Nyssomyia whitmani. Embora nenhuma fêmea tenha sido encontrada naturalmente infectada com flagelados pela dissecação, o fragmento de DNA de Leishmania (Viannia) foi mostrado por multiplex PCR em uma amostra de Ny. neivai (0,46%) e três amostras de Ny. whitmani (1,12%). Conclui-se que Ny. neivai e Ny. whitmani são suscetíveis à infecção por Leishmania, e que multiplex PCR, devido à sua sensibilidade, especificidade e viabilidade, pode ser utilizada em estudos epidemiológicos para a detecção da infecção natural do inseto vetor.

Sandfly frequency in a clean and well-organized rural environment in the state of Paraná, Brazil / Frequência de flebotomíneos em ambiente rural com boas condições de limpeza e organização, no estado do Paraná, Brasil

INTRODUCTION: Sandflies caught in Santa Juliana Farm in Sarandi, State of Paraná, Brazil, were assessed in terms of their fauna, seasonality, and frequency in the homes and in shelters of domestic animals around the homes, as well as in the nearby forest. METHODS: In Santa Juliana Farm, there are no records of cases of ACL, differing from other relatively clean and organized areas where surveys of sandflies have been conducted in Paraná. Samples were collected with Falcão light traps, fortnightly from 22:00 to 02:00 hours, from November 2007 to November 2008. RESULTS: A total of 4,506 sandflies were captured, representing 13 species, predominantly Nyssomyia whitmani (71.8 percent). More sandflies were collected in the forest (52.6 percent) than outside the forest (residences and pigsty) (47.4 percent). However, Ny. whitmani was collected in greater numbers outside (38.3 percent) than inside the forest (33.5 percent). Most sandflies were collected in the warmer months and during periods with regular rainfall. CONCLUSIONS: The results suggest that cleaning and organization around the houses could reduce sandfly population in peridomicile. Constructing shelters for animal at a distance of approximately 100m from domiciles is recommended to prevent the invasion of sandflies, as this farm has an area of preserved forest, with wild animals and sandflies present to maintain the enzootic cycle of Leishmania.

INTRODUÇÃO: Relatam-se os resultados de coletas de flebotomíneos na Fazenda Santa Juliana, município de Sarandi, Estado do Paraná, Brasil, onde se verificou a fauna, a sazonalidade e a frequência desses insetos nos ambientes florestal e extraflorestal. MÉTODOS: O trabalho foi realizado nesta fazenda pela inexistência de registro de casos de LTA e por diferir de outras áreas onde foram realizadas pesquisas com flebotomíneos, no Paraná, e pela limpeza e organização do peridomicílio. As coletas foram realizadas com sete armadilhas luminosas de Falcão, quinzenalmente das 22 às 2 horas, de novembro de 2007 a novembro de 2008. RESULTADOS: Foram capturados 4.506 flebotomíneos de 13 espécies, com predomínio de Nyssomyia whitmani (71,8 por cento). Coletou-se maior proporção de flebotomíneos no ambiente florestal (52,6 por cento) do que no extraflorestal (residências e chiqueiro) (47,4 por cento), entretanto a proporção de Ny. whitmani coletada foi maior no ambiente extraflorestal (38,3 por cento) do que no florestal (33,5 por cento). A maioria dos flebotomíneos foi coletada nos meses mais quentes e com quedas regulares de chuva. CONCLUSÕES: Os resultados sugerem que a limpeza e a organização do peridomicílio podem auxiliar a reduzir a população de flebotomíneos no domicílio e no peridomicílio, recomendando-se que a construção de abrigos de animais guarde a distância aproximada de 100m das residências, para evitar a invasão desses insetos, uma vez que nesta fazenda existe uma área de mata de preservação, com a presença de animais silvestres e flebotomíneos, que mantem o ciclo enzoótico de Leishmania.

Memória espacial e/ou olfativa em flebotomíneos em área endêmica de leishmaniose tegumentar americana, sul do Brasil / Spatial and/or olfactory memory in sandflies in an endemic area for American cutaneous leishmaniasis, southern Brazil

Relata-se os resultados de uma investigação sobre a possibilidade da existência de memória em flebotomíneos em área endêmica de leishmaniose tegumentar americana, no Estado do Paraná. Capturaram-se flebotomíneos no Recanto Marista, Doutor Camargo, Estado do Paraná, Brasil, com armadilhas de Falcão, de 15/11/2007 a 26/11/2007, em dois galinheiros (G1 e G2). Foram capturados 2.080 flebotomíneos (1.000 em G1 e 1.080 em G2), que foram marcados e soltos. Nyssomyia neivai foi a espécie mais (90,5 por cento) freqüente. Do total solto, recapturaram-se 168 (8 por cento) flebotomíneos e a recaptura no G2 foi significativa. Os resultados evidenciam que é possível a existência de memória espacial, olfativa e/ou a fidelidade ao hospedeiro e que isto orientem os flebotomíneos no reconhecimento dos locais onde há disponibilidade de fontes de sangue.

The results from an investigation on the possibility that sandflies in an endemic area for American cutaneous leishmaniasis, in the state of Paraná, may have memory are reported. Sandflies were caught in Recanto Marista, Doutor Camargo, State of Paraná, Brazil, using Falcão traps in two chicken sheds (G1 and G2), between November 15 and 26, 2007. A total of 2,080 sandflies were caught (1,000 in G1 and 1,080 in G2) and these were marked and released. Nyssomyia neivai was the most (90.5 percent) frequent species. Out of the total released, 168 sandflies (8 percent) were recaptured and the recapture rate in G2 was significant. The results show that it is possible that spatial or olfactory memory and/or host loyalty exists, and that this will guide the sandflies in recognizing the places where sources of blood are available.

Investigation of natural infection by Leishmania in sandflies of Paraná State, Southern Brazil

The purpose of this work was to verify the occurrence of Leishmania in naturally infected sandflies. The insects were collected with Falcão, Shannon and HP light-traps, in Doutor Camargo and Maringá municipalities between November 2004 and October 2005. Of the 11,033 sandflies collected in Doutor Camargo, 2,133 surviving females were dissected, particularly those of the Nyssomyia neivai species (86.87 percent). In Maringá, 136 sandflies were collected, of which 79 N. whitmani females and 1 Migonemyia migonei female were dissected. The dissected insects were identified and stored in the pools of 10 specimens. The PCR was carried out on 1,190 females of N. neivai and 190 of N. whitmani from Doutor Camargo, and on 30 of N. whitmani from Maringá, using the primers MP1L/MP3H. The natural infection by Leishmania in sandflies was not confirmed by either of the methods used. The results suggested the low natural infection rate of sandflies by Leishmania in these areas, corroborating other studies carried out in endemic areas of ACL.

Procurou-se verificar a ocorrência de flebotomíneos naturalmente infectados por Leishmania. Estes insetos foram coletados com armadilhas luminosas de Falcão, Shannon e HP, nos municípios de Doutor Camargo e Maringá, de novembro de 2004 a outubro de 2005. Coletaram-se 11.033 flebotomíneos em Doutor Camargo e destes 2.133 fêmeas foram dissecadas, especialmente de Nyssomyia neivai (86,87 por cento). Em Maringá coletaram-se 136 flebotomíneos, dos quais foram dissecadas 79 fêmeas de N. whitmani e 1 Migonemyia migonei. Parte das fêmeas dissecadas foi armazenada em pools de 10 espécimes. A PCR, utilizando os iniciadores MP1L/MP3H, foi realizada em 1.190 fêmeas de N. neivai e 190 de N. whitmani de Doutor Camargo e 30 N. whitmani de Maringá. Não se constatou a infecção natural por Leishmania pelos métodos utilizados. Estes resultados indicam que é baixa a taxa de infecção natural de flebotomíneos por Leishmania, corroborando outros trabalhos realizados em áreas endêmicas de LTA, inclusive no Paraná.

Avaliação de medidas de controle de flebotomíneos / Evaluation of sandfly control measures

Realizou-se coletas de flebotomíneos de maio de 2005 a abril de 2006. Os resultados foram comparados com os das coletas realizadas entre abril de 2001 e setembro de 2002, para avaliar as medidas empregadas para diminuir a densidade destes insetos, no Recanto Marista, município de Doutor Camargo, Estado do Paraná. As coletas foram feitas com armadilhas de Falcão, em domicílios e galinheiros, das 22 às 2 horas, semanalmente, quatro vezes ao mês. Em 2005 e 2006, coletaram-se 213.195 flebotomíneos, 1.113,8 em média por hora (MH) e em 2001 e 2002, 199.821 (MH=1.653,5). Nyssomyia neivai prevaleceu (75,4 por cento) em todos os ecótopos que, juntamente com Nyssomyia whitmani (23,4 por cento), Migonemyia migonei (0,8 por cento) e Pintomyia fischeri (0,4 por cento), representaram 99,7 por cento do total coletado. Nos ecótopos representados por galinheiros foram coletados 88,7 por cento dos flebotomíneos. Constatou-se que houve queda na densidade de flebotomíneos no período de coletas 2005 e 2006 em relação à de 2001 e 2002, especialmente no domicílio.

Collections of sandflies were made between May 2005 and April 2006. The results were compared with those from collections undertaken between April 2001 and September 2002, in order to evaluate the measures used to decrease the density of these insects in Recanto Marista, municipality of Doutor Camargo, State of Paraná. The collections were carried out by Falcão traps inside domiciles and hen sheds, from 10 p.m. to 2 a.m. once a week, four times a month. In 2005 and 2006, 213,195 sandflies were collected (average of 1,113.8 per hour), compared with 199,821 (average of 1,653.5 per hour) in 2001 and 2002. Nyssomyia neivai predominated (75.4 percent) in all the ecotopes. Nyssomyia neivai, Nyssomyia whitmani, Migonemyia migonei and Pintomyia fischeri accounted for 99.7 percent of all the sandflies collected. The ecotope of hencoops accounted for 88.7 percent of the sandflies collected. It was observed that the sandfly density had decreased between the 2001-2002 and 2005-2006 collections, especially in homes.