Felipe Dutra Rêgo

Wild, synanthropic and domestic hosts of Leishmania in an endemic area of cutaneous leishmaniasis in Minas Gerais State, Brazil

Domestic, synanthropic and wild hosts of Leishmania spp. parasites were studied in an area endemic for American tegumentary leishmaniasis (ATL), specifically in northern Minas Gerais State, Brazil. Domestic dogs and small forest mammals are reservoir hosts for L. (Leishmania) infantum. However, the role that these animals play in the transmission cycle of the Leishmania spp. that cause cutaneous leishmaniasis is not well known. This study evaluated 72 rodents, 25 marsupials and 98 domestic dogs found in two villages of the Xakriabá Indigenous Territory, an area of intense ATL transmission. A total of 23 dogs (23.47%) were shown to be positive according to at least one test; 8 dogs (8.16%) tested positive in a single serological test and 15 dogs (15.31%) tested positive by IFAT and ELISA. Eleven dogs were euthanised to allow for molecular diagnosis, of which nine (81.8%) tested positive by PCR for Leishmania in at least one tissue. Seven animals were infected only with L. (L.) infantum, whilst two displayed a mixed infection of L. (L.) infantum and L. (V.) braziliensis. Isoenzymatic characterisation identified L. (L.) infantum parasites isolated from the bone marrow of two dogs. Of the 97 small mammals captured, 24 tested positive for Leishmania by PCR. The results showed that L. (V.) braziliensis, L. (L.) infantum and L. (V.) guyanensis are circulating among wild and synanthropic mammals present in the Xakriabá Reserve, highlighting the epidemiological diversity of ATL in this region.

Ecological aspects of the Phlebotominae fauna (Diptera: Psychodidae) in the Xakriabá Indigenous Reserve, Brazil

BackgroundSand fly collections were performed to study ecological aspects of the Phlebotominae fauna of the Xakriabá Indigenous Reserve, an area with endemic cutaneous leishmaniasis, located in the state of Minas Gerais, Brazil.MethodsThe collections were performed in peridomicile areas and along trails previously selected for the study of wild and synanthropic Leishmania hosts. Differences in the distribution patterns of the sand fly species as well as in species richness and abundance between the different ecotopes were investigated during both rainy and dry seasons over the course of the study period.ResultsA total of 8,046 sand flies belonging to 11 genera and 28 species were collected. Lutzomyia longipalpis and Nyssomyia intermedia were the most abundant species in peridomicile areas, whereas Martinsmyia minasensis and Lutzomyia cavernicola were the most abundant species among the different trail ecotopes.ConclusionThe different composition of the sand fly fauna observed in the peridomicile areas and in the trails during the study, reinforces the importance of sampled different areas in a phlebotomine fauna survey. The presence of Lutzomyia longipalpis and Ny. Intermedia most abundant in peridomicile can be important to Leishmania infantum and Leishmania braziliensis transmission in the Imbaúbas native village.

Molecular Detection of Leishmania in Phlebotomine Sand Flies (Diptera: Psychodidae) from a Cutaneous Leishmaniasis Focus at Xakriabá Indigenous Reserve, Brazil

Autochthonous cases of American cutaneous leishmaniasis (ACL) have been reported since 2001 in the Xakriabá Indigenous Reserve located in the municipality of São João das Missões in northern Minas Gerais state, Brazil. In order to study the presence of Leishmania DNA in phlebotomine sand flies, six entomological collections were carried out from July 2008 through July 2009, using 40 light traps placed in peridomicile areas of 20 randomly selected houses. From October 2011 through August 2012, another six collections were carried out with 20 light traps distributed among four trails (five traps per trail) selected for a previous study of wild and synanthropic hosts of Leishmania. A total of 4,760 phlebotomine specimens were collected belonging to ten genera and twenty-three species. Single female specimens or pools with up to ten specimens of the same locality, species and date, for Leishmania detection by molecular methods. Species identification of parasites was performed with ITS1 PCR-RFLP using HaeIII enzyme and genetic sequencing for SSU rRNA target. The presence of Leishmania DNA was detected in eleven samples from peridomicile areas: Lu. longipalpis (two), Nyssomyia intermedia (four), Lu. renei (two), Lu. ischnacantha, Micropygomyia goiana and Evandromyia lenti (one pool of each specie). The presence of Leishmania DNA was detected in twelve samples from among the trails: Martinsmyia minasensis (six), Ny. intermedia (three), Mi. peresi (two) and Ev. lenti (one). The presence of Leishmania infantum DNA in Lu. longipalpis and Leishmania braziliensis DNA in Ny. intermediasupport the epidemiological importance of these species of sand flies in the cycle of visceral and cutaneous leishmaniasis, respectively. The results also found other species associated with Leishmania DNA, such as Mt. minasensis and Ev. lenti, which may participate in a wild and/or synanthropic cycle of Leishmania transmission in the studied area.

Molecular Detection of Leishmania in Sand Flies (Diptera: Psychodidae: Phlebotominae) Collected in the Caititu Indigenous Reserve of the Municipality of Lábrea, State of Amazonas, Brazil

ABSTRACT Phlebotominae sand flies are of medical importance because they are vectors of human pathogens, such as protozoa of the genus Leishmania Ross, etiological agent of American cutaneous leishmaniasis (ACL). In Lábrea, a municipality in the state of Amazonas, Brazil, ACL is primarily associated with subsistence activities, such as collection and extraction of forest products, undertaken by both indigenous and nonindigenous people. Data on ACL in indigenous populations are scarce, such that there is little information on the identity of the etiologie agent(s), reservoir host(s) and insect vector(s). The aim of this work was to study the sand fly fauna collected during an 8-d surveillance of different habitats in the Indigenous Reserve Caititu, Lábrea. In total, 1,267 sand flies were collected in different habitats for eight consecutive days, of which 819 (64.6%) were females and 448 (35.4%) males, from 10 genera and 32 species. The most abundant genera were Psychodopygus (34.3%), Trichophoromyia (22.9%), and Nyssomyia (15.3%). The most abundant species were Trichophoromyia ubiquitalis (Mangabeira) (n = 235, 18.5%), Psychodopygus davisi (Root) (n = 228, 18.0%) and Nyssomyia antunesi (Coutinho) (n = 135, 10.7%). Direct sequencing of polymerase chain reaction products demonstrated the presence of Leishmania (Leishmania) amazonensis and Leishmania (Viannia) braziliensis in the following species of sand flies: Evandromyia apurinan (Shimabukuro, Silveira, & Silva), Nyssomyia umbratilis (Ward & Fraiha), Nyssomyia yuilli yuilli (Young & Porter), Ps. davisi, Sciopemyia servulolimai (Damasceno & Causey), and Th. ubiquitalis. The presence of natural infection by Leishmania detected in the sand fly species investigated in this study suggests their possible role in the transmission cycle of ACL in the studied area.

Survey of Sand Flies (Diptera: Psychodidae) in an Environmentally Protected Area in Brazil

Brazil is one of the most important endemic areas for leishmaniasis worldwide. Protected areas that are tourist attractions likely present an important risk of transmission of cutaneous leishmaniasis (CL). Furthermore, with the geographical expansion of visceral leishmaniasis (VL), several studies have recorded the occurrence of its vector, Lutzomyia longipalpis, and cases of human and canine VL in such tourist areas. The Parque Estadual do Sumidouro is an environmentally protected area located in the Brazilian Cerrado biome and in an important area endemic for leishmaniasis in the state of Minas Gerais. The purpose of this study was to monitor the sand fly fauna in areas of tourist activity in the park. Sampling was performed every month, from September 2011 to August 2013, using CDC light traps at six sites of differing environmental characteristics. Sampled specimens were identified following Galati (2003), and females were submitted to molecular techniques for the detection and identification of Leishmania DNA. A total of 4,675 sand fly specimens of 25 species belonging to nine genera were collected. The most abundant species were Micropygomyia quinquefer, Lutzomyia renei and Pintomyia pessoai, although only Pi. pessoai is implicated in the transmission of Leishmania braziliensis. The species accumulation curve reached saturation on the 16th sampling event. Species richness, diversity and evenness differed among the sampled areas. The seasonal curve was not determined by a single unique species, and no single species was the most abundant in all environments sampled. The main vector of Leishmania (Leishmania) infantum, Lutzomyia longipalpis, accounted for only 5.35% of the specimens collected. Proven or suspected vectors of Leishmania (Viannia) braziliensis were recorded, and one female of the cortellezzii complex tested positive for Le. braziliensis DNA. Even with a low infection rate (0.62%), these data indicate the circulation of the parasite and reinforce the need for entomological and epidemiological surveillance in the park and its surroundings.

Seasonality of sand flies (Diptera: Psychodidae) and Leishmania DNA detection in vector species in an area with endemic visceral leishmaniasis

BACKGROUND Leishmaniases are a serious health problem in southeast Brazil, including the city of Belo Horizonte (BH), Minas Gerais state (MG), where there are high rates of incidence and mortality due to visceral leishmaniases. BH is divided into nine sanitary districts (SD) of which one, the Venda Nova SD, was selected for this study because it has high rates of positivity for canine leishmaniasis and high incidence of human leishmaniasis. OBJECTIVES This study aimed to survey the sand fly fauna in Venda Nova SD from August 2011 to July 2013 and perform a descriptive analysis of the vector population. METHODS The sampling was carried out using automatic HP light traps at all covered areas of the Venda Nova SD, in a total of eighteen light traps. Sampled specimens were identified following Galati (2003), and females were submitted to molecular techniques for the detection and identification of Leishmania DNA. A simple environmental description was done for it area and Kernel estimation was used to infer vector density for each study site. FINDINGS A total of 2,427 sand fly specimens belonging to eight species and five genera were collected of which 95.3% were Lutzomyia longipalpis. The seasonal variation curve was delineated by this species. Lu. longipalpis was the most abundant at all collection points and in all months of the study, and exhibited a natural infection rate of 1.01% for Leishmania infantum and 1.77% for Leishmania braziliensis. MAIN CONCLUSIONS The results show the presence and adaptation of Lu. longipalpis to the anthropic environment of BH and reinforces its role as the main vector of L. infantum in the region.

Molecular Detection of Leishmania DNA in Wild-Caught Phlebotomine Sand Flies (Diptera: Psychodidae) From a Cave in the State of Minas Gerais, Brazil

Abstract Leishmania spp. are distributed throughout the world, and different species are associated with varying degrees of disease severity. In Brazil, Leishmania transmission involves several species of phlebotomine sand flies that are closely associated with different parasites and reservoirs, and thereby giving rise to different transmission cycles. Infection occurs during the bloodmeals of sand flies obtained from a variety of wild and domestic animals, and sometimes from humans. The present study focused on detection of Leishmania DNA in phlebotomine sand flies from a cave in the state of Minas Gerais. Detection of Leishmania in female sand flies was performed with ITS1 PCR-RFLP (internal transcribed spacer 1) using HaeIII enzyme and genetic sequencing for SSUrRNA target. The survey of Leishmania DNA was carried out on 232 pools and the parasite DNA was detected in four: one pool of Lutzomyia cavernicola (Costa Lima, 1932), infected with Le. infantum (ITS1 PCR-RFLP), two pools of Evandromyia sallesi (Galvão & Coutinho, 1939), both infected with Leishmania braziliensis complex (SSUrRNA genetic sequencing analysis), and one pool of Sciopemyia sordellii (Shannon & Del Ponte, 1927), infected with subgenus Leishmania (SSUrRNA genetic sequencing analysis). The present study identified the species for Leishmania DNA detected in four pools of sand flies, all of which were captured inside the cave. These results represent the first molecular detection of Lu. cavernicola with Le. infantum DNA, Sc. sordellii with subgenus Leishmania DNA, and Ev. sallesi with Leishmania braziliensis complex DNA. The infection rate in females captured for this study was 0.17%.

Description of the Female of Martinsmyia minasensis (Diptera, Psychodidae, Phlebotominae), With Distribution Records and a Key to Female Identification.

Abstract The adult female of the sand fly species Martinsmyia minasensis (Mangabeira, 1942) is described, and the characters of the genus Martinsmyia Galati, 1995 are defined. We performed morphometric analyses on the spermatheca and wings, and present data on the geographical distribution of M. minasensis, as well as an identification key to females of this genus. The specimens were measured, drawn, photographed, and compared with the similar species M. oliveirai. It was possible to distinguish between M. minasensis and M. oliveirai based on differences found in the common and individual ducts of the spermatheca, as well as wing shape and size.

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

BackgroundPhlebotomines 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.MethodsPhlebotomines 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.ResultsA 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.ConclusionThe 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.

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.