Sand fly bioecological aspects and risk mapping of leishmaniasis by geographical information systems approach in a mineral exploration area of Brazil.

Epidemiological studies of leishmaniasis in areas of great human influence and environmental change serve as important tools for the implementation of effective control plans. Mining is currently a major economic activity in Brazil with the municipality of Pains, in the state of Minas Gerais, being one of the main lime producing municipalities in the country. This study aimed to map areas of potential transmission risks within the municipality of Pains using an epidemiological approach in association with the ecological study of sand flies. Twelve samplings carried out between May 2015 and April 2016 collected a total of 12,728 sandflies, comprising 2,854 females (22.42%) and 9,874 males (77.58%), of 20 species belonging to ten genera. The most abundant species was Lutzomyia longipalpis (80%). Leishmania DNA was detected in seven pools of female sand flies with an infection rate of 0.37%. Geoprocessing and the use of maps revealed that vector sand flies are distributed throughout the urban area, as are cases of canine and human leishmaniasis. However, the greatest abundances of sand flies were at sampling points at the border of the urban area. Higher densities of sand flies and the presence of Leishmania DNA may be correlated with extensive degradation by limestone mining. Integrated and multidisciplinary research approaches are necessary to better understand how the impacts of environmental change influence these insect vectors of leishmaniasis.

Absence of yellow fever virus circulation in wildlife rodents from Brazil.

Yellow fever (YF), caused by the yellow fever virus (YFV), is an emerging viral zoonosis that affects humans and non-human primates (NHP). In South America, YF is naturally maintained through enzootic/sylvatic cycles involving NHPs and mosquitoes (Haemagogus and Sabethes). In this study, we retrospectively analyzed wildlife rodents to better understand their role in a potential alternative YF sylvatic cycle. The plaque reduction neutralization test was performed to detect anti-YFV antibodies, while qPCR targeting the NS5 region of flaviviruses and standard PCR targeting the CprM region were applied to detect YFV RNA in tissue and blood samples. YFV was not evidenced in any of the tested samples. These findings provide additional information regarding sylvatic YFV and emphasize the importance of YFV surveillance in wild animals as potential reservoirs/hosts given the well-established enzootic cycle in the studied areas, mainly in the Atlantic Forest.

Sand fly behavior: much more than weak-flying.

BACKGROUND: Leishmaniases are diseases transmitted by some species of sand flies and are widely distributed throughout the tropical regions of the planet. Despite the low mobility of these vectors, the geographical distributions of some species are quite extensive, which hinders control and surveillance measures in endemic areas. OBJECTIVES: The present study investigated the flying capacity of sand flies. METHODS: Four Hoover Penido (HP)-type light traps were positioned in the centre of the Velhas' River, about 80 metres equidistant from each other. We also realised capture/release/recapture attempts to assess possible capacity of phlebotomine fly uninterrupted up to 150 metres. Captured sand flies from one side of the river were marked using fluorescent powder (Luminous Paint kit, Bioquip®) and released on the other side, approximately 150 m distant. Recapture attempts were made on river's bank up to 30 days post-release. FINDINGS: Six sand flies of the species Nyssomyia neivai (n = 4), Ny. intermedia (n = 1) and Evandromyia lenti (n = 1) were captured in the centre of the river. There were no recaptures of the 1,450 marked-and-released sand flies. MAIN CONCLUSIONS: The results obtained disagree with data found in the literature regarding the flight capacity of sand fly vectors of leishmaniasis.

The sand fly (Diptera: Psychodidae) fauna of the urban area of Lassance, Northeast Minas Gerais, Brazil.

The present study aimed to check the sand flies' fauna on the municipality of Lassance, Minas Gerais, Brazil and detect the presence of Leishmania DNA on the female captured and determine the risk areas of the municipality. Sand flies were collected monthly from May 2018 to April 2019 using automatic light traps for 3 consecutive nights. Eight houses were selected as sample points due its previous reports of tegumentary leishmaniasis and/or canine leishmaniasis. The sand fly's fauna found on the present study it's represented by several medical importance species and the most abundant species found were Lutzomyia longipalpis (77.09%) and Nyssomyia intermedia (10.06%). Leishmania infantum DNA was detected in a pool of Lu. longipalpis resulting on a 2.81% of infection rate. By the frequency of the two most abundant species on this study, we developed a risk area map and it draws attention to sample point 6 due to disparate abundance of sand flies at this site (81.81%). Statistical overview shows Lu. longipalpis as dominant species and, still, Non-Metric Multidimensional Scaling analysis reveal high similarity on fauna's diversity on the study area. Our findings suggest that the diversity of sand flies from the municipality of Lassance may promote the circulation of Leishmania infantum parasites putting in risk the habitants and other mammal's species. Still, our study reinforces the necessity of specific studies focused on breed sites of phlebotomine and its' ecology to expand the knowledge about the behaviour of this group of insects applying directly to leishmaniases' epidemiology.

Sand fly behavior: much more than weak-flying

BACKGROUND Leishmaniases are diseases transmitted by some species of sand flies and are widely distributed throughout the tropical regions of the planet. Despite the low mobility of these vectors, the geographical distributions of some species are quite extensive, which hinders control and surveillance measures in endemic areas. OBJECTIVES The present study investigated the flying capacity of sand flies. METHODS Four Hoover Penido (HP)-type light traps were positioned in the centre of the Velhas' River, about 80 metres equidistant from each other. We also realised capture/release/recapture attempts to assess possible capacity of phlebotomine fly uninterrupted up to 150 metres. Captured sand flies from one side of the river were marked using fluorescent powder (Luminous Paint kit, Bioquip®) and released on the other side, approximately 150 m distant. Recapture attempts were made on river's bank up to 30 days post-release. FINDINGS Six sand flies of the species Nyssomyia neivai (n = 4), Ny. intermedia (n = 1) and Evandromyia lenti (n = 1) were captured in the centre of the river. There were no recaptures of the 1,450 marked-and-released sand flies. MAIN CONCLUSIONS The results obtained disagree with data found in the literature regarding the flight capacity of sand fly vectors of leishmaniasis.

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.

Detection of Leishmania spp in silvatic mammals and isolation of Leishmania (Viannia) braziliensis from Rattus rattus in an endemic area for leishmaniasis in Minas Gerais State, Brazil.

Knowledge of potential reservoirs of Leishmania spp. in an anthropic environment is important so that surveillance and control measures can be implemented. The aim of this study was to investigate the infection by Leishmania in small mammals in an area located in Minas Gerais, Brazil, that undergoes changes in its natural environment and presents autochthonous human cases of cutaneous leishmaniasis (CL) and visceral leishmaniasis (VL). For the capture of the animals, Sherman and Tomahawk traps were used and distributed in the peridomicile of houses with reports of autochthonous cases of CL or VL. Six catches were carried out on two consecutive nights with intervals of two months during one year and samples of spleen, liver, tail skin, ear skin and bone marrow of the animals were obtained. Parasitological and molecular methods were used to detect the infection. Identification of the Leishmania species was performed by PCR RFLPhsp70. Twenty five animals of four species were captured: ten Rattus rattus, nine Didelphis albiventris, five Cerradomys subflavus and one Marmosops incanus. In the PCR-hsp70, five animals were positive (20%). The Leishmania species identified in PCR-RFLPhsp70 were: Leishmania braziliensis in D. albiventris (2), C. subflavus (1) and R. rattus (1) and Leishmania infantum in R. rattus (1). The highest positivity rate for L. braziliensis was obtained in the liver samples. The spleen was the only tissue positive for L. infantum. It was isolated in culture medium L. braziliensis from two samples (liver and spleen) of R. rattus. This is the first record of isolation of L. braziliensis from R. rattus in the southeastern region of Brazil. These results are relevant to the knowledge of the epidemiology of leishmaniasis in the region, mainly in the investigation of the presence of hosts and possible reservoirs of the parasite.

Abundance, survival, recruitment and effectiveness of sterilization of free-roaming dogs: A capture and recapture study in Brazil.

The existence of free-roaming dogs raises important issues in animal welfare and in public health. A proper understanding of these animals' ecology is useful as a necessary input to plan strategies to control these populations. The present study addresses the population dynamics and the effectiveness of the sterilization of unrestricted dogs using capture and recapture procedures suitable for open animal populations. Every two months, over a period of 14 months, we captured, tagged, released and recaptured dogs in two regions in a city in the southeast region of Brazil. In one of these regions the animals were also sterilized. Both regions had similar social, environmental and demographic features. We estimated the presence of 148 females and 227 males during the period of study. The average dog:man ratio was 1 dog for each 42 and 51 human beings, in the areas without and with sterilization, respectively. The animal population size increased in both regions, due mainly to the abandonment of domestic dogs. Mortality rate decreased throughout the study period. Survival probabilities did not differ between genders, but males entered the population in higher numbers. There were no differences in abundance, survival and recruitment between the regions, indicating that sterilization did not affect the population dynamics. Our findings indicate that the observed animal dynamics were influenced by density-independent factors, and that sterilization might not be a viable and effective strategy in regions where availability of resources is low and animal abandonment rates are high. Furthermore, the high demographic turnover rates observed render the canine free-roaming population younger, thus more susceptible to diseases, especially to rabies and leishmaniasis. We conclude by stressing the importance of implementing educational programs to promote responsible animal ownership and effective strategies against abandonment practices.

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 eco-epidemiológico dos hospedeiros e reservatórios de Leishmania na Reserva Particular do Patrimônio Natural Santuário do Caraça, Minas Gerais

As leishmanioses são doenças parasitárias de origem zoonótica, sendo transmitidas por flebotomíneos vetores em ambiente silvestres, rurais e urbano. Sua distribuição está condicionada não somente ao vetor, mas também aos mamíferos reservatórios. O presente trabalho teve como objetivo realizar estudos eco-epidemiológicos em relação aos hospedeiros vertebrados e invertebrados de Leishmania spp. no Santuário do Caraça. Do período de Junho de 2013 a Junho de 2014 foram realizadas coletas bimestrais intercaladas de flebotomíneos e de pequenos mamíferos em pontos selecionados ao acaso em locais de atração turística na RPPN Santuário do Caraça. Foram utilizadas 25 armadilhas luminosas do tipo CDC para coletas de flebotomíneos distribuídas em sete trilhas pelo parque. Para coleta dos pequenos mamíferos foram utilizadas 60 armadilhas do tipo Tomahalk distribuídas em seis trilhas (dez armadilhas por trilha). Um total de 376 flebotomíneos foi coletado (300 fêmeas e 76 machos) representando 18 espécies.

As espécies mais abundantes foram Psychodopygus lloydi(72,79%), Brumptomyia troglodytes (5,25%), Nyssomyia whitmani (4,01%) e Pintomyia monticola (4,30%). Duas amostras de Ps. lloydi foram detectadas com DNA de Leishmania (Viannia) braziliensis, utilizando a técnica ITS1 PCR-RFLP, e sete espécimes desta espécie foram identificados com sangue de porco doméstico (Sus scrofa) através de sequenciamento do gene ctyb. Um total de 55 pequenos mamíferos foi coletado tendo como espécies mais abundantes Akodon cursor (56,4%), Cerradomys subflavus (10,9%) e Oligoryzomys nigripes(10,9%). Foram obtidos seis isolados de Le. (V.) braziliensis de fígado e pele de cauda dos mamíferos das espécies A. cursor (3), C. subflavus (1) e Oxymicterus dasytrichus (1). Estes mesmos espécimes foram detectados positivos na técnica de kDNA PCR. Os resultados encontrados inferem que há circulação de Le. (V.) braziliensis neste ambiente silvestre e que Ps. lloydi possa desempenhar papel importante na manutenção do ciclo neste local e que, ainda, A. cursor, C. subflavus e O. dasytrichus atuam como hospedeiros de leishmaniose tegumentar na área de estudo

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.

Molecular detection of Leishmania in phlebotomine sand flies (Diptera: Psychodidae) from a cutaneous leishmaniasis focus atXakriabá 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.

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

BACKGROUND: Sand 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. METHODS: The 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. RESULTS: A 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. CONCLUSION: The 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.

Study of sand flies (Diptera: Psychodidae) in visceral and cutaneous leishmaniasis areas in the central-western state of Minas Gerais, Brazil.

The transmission of Leishmania involves several species of sand flies that are closely associated with various parasites and reservoirs, with differing transmission cycles in Brazil. A study on the phlebotomine species composition has been conducted in the municipality of Divinópolis, Minas Gerais, Brazil, an endemic area for cutaneous leishmaniasis (CL), which has intense occurrence of visceral leishmaniasis (VL) cases. In order to study the sand flies populations and their seasonality, CDC light traps (HP model) were distributed in 15 houses which presented at least one case of CL or VL and in five urban parks (green areas). Collections were carried out three nights monthly from September 2010 to August 2011. A total of 1064 phlebotomine specimens were collected belonging to two genera and seventeen species: Brumptomyia brumpti, Lutzomyia bacula, Lutzomyia cortelezzii, Lutzomyia lenti, Lutzomyia sallesi, Lutzomyia longipalpis, Lutzomyia migonei, Lutzomyia intermedia, Lutzomyia neivai, Lutzomyia whitmani, Lutzomyia christenseni, Lutzomyia monticola, Lutzomyia pessoai, Lutzomyia aragaoi, Lutzomyia brasiliensis, Lutzomyia lutziana, and Lutzomyia sordellii. L. longipalpis, the main vector of Leishmania infantum in Brazil, was the most frequent species, accounting for 76.9% of the total, followed by L. lenti with 8.3%, this species is not a proven vector. Green and urban areas had different sand flies species composition, whereas the high abundance of L. longipalpis in urban areas and the presence of various vector species in both green and urban areas were also observed. Our data point out to the requirement of control measures against phlebotomine sand flies in the municipality of Divinópolis and adoption of strategies aiming entomological surveillance.