Diversity of phlebotomine sand flies and molecular detection of trypanosomatids in Brumadinho, Minas Gerais, Brazil.

This study aimed to describe the sand fly fauna and detect trypanosomatids in these insects from Casa Branca, state of Minas Gerais, Brazil, an endemic area of both visceral (VL) and tegumentary leishmaniasis (TL). Sand flies were collected bimonthly from May 2013 to July 2014, using automatic light traps exposed for three consecutive nights in peridomiciliary areas of nine houses with previous reports of VL and TL. ITS1-PCR and DNA sequencing were performed for trypanosomatids identification. A total of 16,771 sand flies were collected belonging to 23 species. The most abundant species was Nyssomyia whitmani (Antunes & Coutinho, 1939) (70.9%), followed by Lutzomyia longipalpis (Lutz & Neiva, 1912) (15.2%) and Migonemyia migonei (França, 1920) (9.1%). Leishmania amazonensis DNA was detected in Ny. whitmani (four pools) and Le. braziliensis DNA was detected in Psychodopygus lloydi (one pool). In seven pools of Ny. whitmani and in one pool of Lu. longipalpis positive for Leishmania DNA, the parasite species was not determined due to the low quality of the sequences. Moreover, DNA of Herpetomonas spp. was detected in Ny. whitmani (two pools) and Cortelezzii complex (one pool). DNA of Crithidia spp. was detected in Ny. whitmani and Ps. lloydi (both one pool). Our results suggest that Ny. whitmani may be involved in the transmission of Le. amazonensis in the study area. The molecular detection of Le. amazonensis suggests the presence of this species in a sylvatic cycle between vertebrate and invertebrate hosts in the region of Casa Branca. Our data also reveal the occurrence of other non-Leishmania trypanosomatids in sand flies in Casa Branca District.

Leishmania (Viannia) braziliensis infection in wild small mammals in ecotourism area of Brazil.

Leishmaniases are parasitic diseases transmitted to mammalian hosts by sand fly vectors (Diptera: Psychodidae). Despite the increasing occurrence of visceral and cutaneous leishmaniasis cases in urban centers, their transmission still occur primarily in wild environments and may be associated with professional activities and recreation, such as ecotourism. The Reserva Particular do Patrimônio Natural Santuário do Caraça (RPPNSC) is one of the largest ecotourism attractions in the State of Minas Gerais, Brazil, and comprises an area of environmental preservation with 11,233 hectares presenting a transitional vegetation between Cerrado and Atlantic Forest. The present study describes the abundance of small mammals in RPPNSC, the isolation and identification of Leishmania in five wild animals. Small mammals were bimonthly trapped along 6 trails within the RPPNSC with 10 Tomahawk traps each. Two trails were located in peridomiciliary areas near tourist lodging facilities, and four trails were located at sites visited by tourists in forest areas. The most prevalent species were Akodon cursor, Cerradomys subflavus and Oligoryzomys nigripes. Six isolates of Leishmania were obtained from these animals and identified as Leishmania braziliensis through HSP70-PCR RFLP method. Leishmania spp. DNA was detected by kDNA-PCR method and isolated by biphasic culture. Studies point to some of the captured species as potential wild reservoirs of Leishmania, suggesting they may be involved in the transmission cycle in these wild environments.

Aspects of the ecology of phlebotomine sand flies (Diptera: Psychodidae) in the Private Natural Heritage Reserve Sanctuary Caraça.

Leishmaniases are a set of parasitic diseases of zoonotic origin that are transmitted by sandfly vectors in wild, rural and urban environments. Their distribution is dependent not only the distribution of vectors, but also on the distribution of mammalian reservoirs. Only by understanding the transmission cycle of these diseases, such as knowing the participating vectors and reservoirs, can one can understand the epidemiology and ecological relationships of leishmaniases. Ecotourism has become an important area of economic growth in Brazil. One of the most visited tourist attractions in the state of Minas Gerais, the Reserva Particular do Patrimônio Natural Santuário do Caraça (RPPNSC) is located in the Quadrilátero Ferrífero. The aim of this study was to contribute to the control of leishmaniasis among tourists of the RPPNPC by surveying its sand fly fauna and testing for the presence of Leishmania DNA in females. Twenty-five CDC light traps were exposed on 7 trails of the RPPNPC where samples were collected bimonthly for a year, starting in June 2013. A total of 376 specimens of 18 species and 10 genera of sandflies were captured. The predominant species were Psychodopygus lloydi (72.34%) and Pintomyia monticola (5.59%). HaeIII restriction enzyme detected and characterized Leishmania braziliensis DNA in 2 of the samples for an infection rate of 0.7% (2/266). Recent studies found specimens of Ps. lloyd infected with Leishmania braziliensis elsewhere in Minas Gerais, which may be an indication that this species is involved in the transmission of Leishmania in this state.

Estudo dos flebotomíneos (Diptera: Psychodidae) na localidade de Casa Branca, município de Brumadinho, área de transmissão para leishmanioses no Estado de Minas Gerais

As leishmanioses são doenças endêmicas em vários países do mundo, incluindo o Brasil e são causadas por parasitas do gênero Leishmania. O conhecimento sobre os vetores dessas doenças, os flebotomíneos, pode auxiliar no delineamento das medidas de controle da doença em determinados locais. O objetivo deste trabalho foi estudar a fauna de flebotomíneos, verificar a presença de DNA de Leishmanianas fêmeas capturadas e verificar a alimentação sanguínea nas fêmeas ingurgitadas coletadas na localidade de Casa Branca, pertencente ao município de Brumadinho, Minas Gerais. Durante o período de maio de 2013 a julho de 2014 foram realizadas coletas bimensais totalizando oito coletas sistematizadas de flebotomíneos utilizando 18 armadilhas luminosas expostas no peridomicílio de nove casas, selecionadas por apresentarem casos de leishmanioses humana ou canina. A fauna de flebotomíneos foi composta por 23 espécies, com um total de 16.771 flebotomíneos capturados, sendo a espécie Nyssomyia whitmani a mais abundante na área de estudo, seguida por Lutzomyialongipalpis e Migonemyiamigonei. As fêmeas não alimentadas pertencentes às coletas dos meses de maio/2013, setembro/2013, janeiro/2014 e maio/2014 foram dissecadas e colocadas em pools de no máximo 20 indivíduos onde foi feita a extração de DNA e posterior pesquisa de DNA de Leishmania por meio da técnica de PCR dirigida ao alvo ITS1.

As espécies de Leishmania e outros tripanossomatídeos detectados nos pools das fêmeas de flebotomíneos neste estudo foram identificados através da técnica de sequenciamento genético do produto amplificado. Foi possível detectar DNA de Leishmania em nove pools: Lu. longipalpis (1), Ny. whitmani (6) e Psychodopygus lloydi (2).Outros tripanossomatídeos foram detectados em 10 pools: Crithidia sp.em 1 pool de Ps. lloydi, Endotrypanum sp. em 6 pools de Ny. whitmani e Herpetomonas sp. nas espécies Ny. whitmani (3 pools) e no complexo cortelezzii (1 pool). As fêmeas alimentadas foram dissecadas e o DNA foi extraído para posterior identificação de fontes alimentares através da PCR dirigida ao gene do citocromo b com a confirmação pelo sequenciamento genético. Foram detectadas as fontes alimentares Canis familiaris, Gallus gallus, Homo sapiens e Rattus rattus, e a espécie Ny. whitmani foi a mais abundante capturada alimentada no peridomicílio. Os resultados da pesquisa de DNA de Leishmania chamam atenção para a presença das espécies de importância médica como Lu. longipalpis e Ny. whitmani presentes na área de estudo detectadas com Leishmania, reforçando seus papéis na epidemiologia das leishmanioses. Os outros tripanossomatídeos detectados nesse estudo mostram que pela PCR-ITS1 foi possível detectar outras espécies da família Trypanosomatidae além de Leishmania, evidenciando que essas espécies estão presentes na área de estudo. Os resultados do estudo das fontes alimentares mostraram que os flebotomíneos estão adaptados ao peridomicílio se alimentando em animais comuns a este ambiente

IDENTIFICATION OF SANDFLIES (Diptera: Psychodidae: Phlebotominae) BLOOD MEALS IN AN ENDEMIC LEISHMANIASIS AREA IN BRAZIL.

The aim of this study was to identify blood meals of female sandflies captured in the municipality of Governador Valadares, an endemic area of visceral and cutaneous leishmaniasis, in the State of Minas Gerais, Brazil. From May 2011 to January 2012, captures were performed using HP light traps in four districts. There were 2,614 specimens (2,090 males and 524 females) captured; 97 engorged females were identified belonging to the species Lutzomyia longipalpis(82.1%) and Lutzomyia cortelezzii(17.9%). Considering simple and mixed feeding, the enzyme-linked immunosorbent assay revealed a predominance of chicken blood (43.6%) in Lutzomyia longipalpis, showing the important role that chickens exert around the residential areas of Governador Valadares. This finding increases the chances of sandflies contact with other vertebrates and consequently the risk of leishmaniasis transmission.

Comparison of the phlebotomine (Diptera: Psychodidae) fauna of urban, transitional, and wild areas in northern Minas Gerais, Brazil.

BACKGROUND: Phlebotomines are directly related to the study of leishmaniases, and so the study of their distribution plays an important role in the epidemiology of these diseases. Collections of phlebotomines were made with the intent of comparing the distribution, richness, diversity, and abundance of species in three distinct environments in an area endemic for tegumentary and visceral leishmaniasis in Minas Gerais State, Brazil. METHODS: Phlebotomines were collected with automatic light traps in urban, transitional, and wild areas from March 2013 to February 2014 in the district of Barra do Guaicuí, municipality of Várzea da Palma, Minas Gerais. The distribution patterns of these species of insects, as well as species richness, evenness, and abundance among the different areas, were analyzed. RESULTS: A total of 3,365 phlebotomines belonging to 15 species were collected. The urban area had the greatest abundance whereas the transitional area had the greatest diversity and evenness of species. Nyssomyia intermedia was the most abundant species in the urban area, whereas Evandromyia evandroi was the most abundant in the transitional area and Ev. lenti in the wild area. CONCLUSION: The analysis of our results showed that the distribution of the collected species had distinct profiles between the environments studied. Furthermore our study indicates the potential risk of transmission of leishmaniasis in the urban environment where it was observed had the highest population density and abundance of important vector species of Leishmania.

Identification of sandflies (Diptera: Psychodidae: Phlebotominae) blood meals in an endemic Leishmaniasis area in Brazil / Identificação do repasto sanguíneo de flebotomíneos (Diptera: Psychodidae: Phlebotominae) provenientes de área endêmica de Leishmaniose no Brasil

SUMMARYThe aim of this study was to identify blood meals of female sandflies captured in the municipality of Governador Valadares, an endemic area of visceral and cutaneous leishmaniasis, in the State of Minas Gerais, Brazil. From May 2011 to January 2012, captures were performed using HP light traps in four districts. There were 2,614 specimens (2,090 males and 524 females) captured; 97 engorged females were identified belonging to the species Lutzomyia longipalpis(82.1%) and Lutzomyia cortelezzii(17.9%). Considering simple and mixed feeding, the enzyme-linked immunosorbent assay revealed a predominance of chicken blood (43.6%) in Lutzomyia longipalpis, showing the important role that chickens exert around the residential areas of Governador Valadares. This finding increases the chances of sandflies contact with other vertebrates and consequently the risk of leishmaniasis transmission.

RESUMOO objetivo deste estudo foi identificar o repasto sanguíneo de fêmeas de flebotomíneos capturadas no município de Governador Valadares, área endêmica de leishmaniose visceral e tegumentar no Estado de Minas Gerais, Brasil. Entre maio de 2011 e janeiro 2012 foram realizadas capturas com armadilhas luminosas HP em quatro bairros. Foram capturados 2.614 exemplares (2.090 machos e 524 fêmeas). Noventa e sete fêmeas ingurgitadas foram identificadas como pertencentes às espécies Lutzomyia longipalpis(82,1%) e Lutzomyia cortelezzii(17,9%). Considerando a alimentação simples e a mista, o ensaio imunoenzimático revelou em Lutzomyia longipalpisuma predominância de sangue de galinhas (43,6%), mostrando o importante papel que galinhas podem exercer no peridomicílio, aumentando a chance de contato dos flebotomíneos com outros vertebrados e, consequentemente, o risco de transmissão da leishmaniose.

Epidemiology of visceral leishmaniasis in a reemerging focus of intense transmission in Minas Gerais State, Brazil.

This study was developed in the urban area of Governador Valadares, a reemerging focus of intense transmission of visceral leishmaniasis (VL) in Brazil, presenting 86 human cases of VL from 2008 to 2011. The disease prevailed in males (73.2%) with most patients between 0 and 9 years (44.1%) and a lethality rate of 16.2%. A canine survey was carried out on 16,529 domestic dogs in 35 districts in the area and it showed that 30.2% of them (4,992 dogs) were positive for VL by serum assays. Prevalence ratios for canine VL varied between 13.6% and 53.4%. The clinical exam of 343 seropositive dogs showed that 49.9% of them were considered symptomatic, with larger prevalence of canine VL being in short-furred animals (90%). The entomological survey was performed in eight districts, where 2,539 phlebotomines were captured, preferentially in the peridomicile (84.5%). Lutzomyia longipalpis was the predominant species (90%) suggesting its participation in the VL transmission in the area. The correlation between canine prevalence and L. longipalpis density was evaluated.