Revisiting the cave-dwelling sand flies (Diptera, Psychodidae, Phlebotominae) from Brazil: Diversity and potential role in the transmission of Leishmania Ross, 1903 (Kinetoplastida: Trypanosomatidae).

Brazilian caves, one of the many tourist attractions of the country, may act as a shelter for insects, such as sand flies (Diptera: Psychodidae), natural hosts of various microorganisms including parasites of the genus Leishmania Ross, 1903. In the last decades, with the increasing global need for sustainable development, ecotourism has emerged as one of the major activities in Brazil. However, the constant monitoring in environmentally protected areas is not often carried out, endangering visitors and professionals, especially due to the occurrence of zoonoses. Several sand fly species have already been recorded in Brazilian caves, drawing attention to the possibility of Leishmania transmission at this ecotope. Indeed, this current systematic review summarizes the fauna of cave-dwelling sand flies in Brazil, focusing on their biological behaviour and the occurrence of potential vectors of Leishmania parasites.

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.

Biodiversity of Hematophagous Diptera (Culicidae and Psychodidae) in a Small Hydroelectric Power Station and Surrounding Area in the State of Minas Gerais, Brazil.

Hydroelectric power stations may affect the population dynamics of mosquitoes and sand flies, a group with impact on public health due to the possibility to transmit pathogens to humans. This work characterized and compared the fauna of mosquitoes and sand flies in a hydroelectric power station and peridomicile areas in the State of Minas Gerais, Brazil. Insect collections were performed in August 2015 at dry season and February 2016 in rainy season. Ten HP light traps were set at each of two sites for three consecutive days in each of two seasons (dry and rainy). Furthermore, collections with Shannon traps were made in each sampling area (hydropower plant and peridomicile area) from 4:00 p.m. being shut down at 8:00 p.m. for two consecutive days in each of two seasons (dry and rainy). In total, 1,222 insects from 13 genera and 27 species were collected. The most plentiful species were respectively Culex declarator (Dyar and Knab, 1906) and Pintomyia pessoai (Coutinho & Barretto, 1940). A high number of insects (78.5%) were collected during the rainy period (P < 0.05). About equitability, statistical significance was observed in the peridomicile area (dry season J = 0.75 and rainy season J = 0.82). The highest values of species diversity were observed in the hydropower plant (H = 2.68) and peridomicile area (H = 2.38) both in the rainy season with statistical significance comparing with dry season. Our results demonstrate that the occurrence of vector species in this region increases the potential risk of transmission of pathogens, especially arbovirus and Leishmania Ross, 1903.

The sand fly species (diptera: psychodidae) in an urban environment of Mato Grosso do Sul, Brazil

The purpose of this study was to search for Lutzomyia longipalpis and Lutzomyia cruzi in the urban area of the Água Clara Municipality, Mato Grosso do Sul State, Brazil. Two CDC traps were placed in the intradomicile and the peridomicile areas in four residences once a month, for three consecutive nights during 24 months. Were collected 11 species of sand flies totaling 3,418 specimens, 2,762 males and 656 females. Lu. longipalpis was the dominant species and there was no statistical difference between the peridomicile and intradomicile areas. Lu. cruzi was not found. The high capture rate for Lu. longipalpis in the urban area of Água Clara is cause for concern regarding necessary preventive measures to avoid future cases of visceral leishmaniasis.

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.

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

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.

Ecological aspects of phlebotomine sandflies (Diptera: Psychodidae) from a cave of the speleological province of Bambuí, Brazil.

Phlebotomines are invertebrate hosts of Leishmania genus species which are etiological agents of leishmaniases in humans and other mammals. Sandflies are often collected in entomological studies of caves both in the inner area and the adjacent environments. Caves are ecotypes clearly different from the external environment. Several caves have been opened to public visitation before any studies were performed and the places do not have scientific monitoring of the fauna, flora, geological and geographical characteristics. These events can lead to the loss of geological and biological information. Considering these aspects, this study aimed to describe the sand fly fauna, including the ecological features, in a limestone cave at the Speleological Province of Bambuí (Minas Gerais State, Brazil). A total of 8,354 specimens of sandflies belonging to 29 species were analyzed: Lutzomyia cavernicola (20%), Nyssomyia intermedia (15%), Martinsmyia oliveirai (13%), Evandromyia spelunca (12%), Evandromyia sallesi (11%), Migonemyia migonei (9%), Nyssomyia whitmani (9%), Sciopemyia sordellii (4%) and Lutzomyia longipalpis (2%). The others species represent 5% of the total. This manuscript presents data found on richness, diversity, evenness and seasonality, comparing the sand fly fauna trapped in the cave and its surroundings.

Anomalies in the sand fly Lutzomyia longipalpis (Diptera: Psychodidae) in Brazil.

Lutzomyia longipalpis is the main species involved in the epidemiological cycle of visceral leishmaniasis in Brazil. This species shows a wide geographic distribution and belongs to a group that has 2 setae in the paramere with the curved tips towards the apex of this structure among other characteristics. Morphological characters in the genitalia of phlebotomines are essential for correct identification of species. The aim of this study was to report the finding of unilateral and bilateral anomalies in the paramere of L. longipalpis males collected in Várzea da Palma municipality, Minas Gerais State, Brazil. Twelve collections were made during the period April 2009 to March 2010, using Centers for Disease Control and Prevention light traps, HP model. Of the 8,832 male L. longipalpis collected, we found 6 types of unilateral anomalies and 2 types of bilateral anomalies in 0.5% of the insects collected. The anomalies were described according to number, position, and size of the setae of paramere. This information will be useful in the validation of new species.

Lutzomyia diamantinensis sp. nov., a new phlebotomine species (Diptera: Psychodidae) from a quartzite cave in Diamantina, state of Minas Gerais, Brazil

A new species of Brazilian phlebotomine sandfly found in Brazil, municipality of Diamantina, state of Minas Gerais, is described based on males and females collected in a quartzite cave. The body of spermathecae is continuous to the individual duct, lanky and tapering at the end, with conical shaped, not striated and presenting the head with dense setae. The male presents gonostyle with four spines and a small subterminal seta and gonocoxite with one group of persistent setae. The paramere is simple with a group of small setae on the dorsal apex. The morphological features of this new species permit its inclusion in the migonei group.

Biogeographical aspects of the occurrence of Nyssomyia neivai and Nyssomyia intermedia (Diptera: Psychodidae) in a sympatric area of the Brazilian savannah.

Nyssomyia intermedia and Nyssomyia neivai constitute a species complex associated with Leishmania transmission. The aim of this study was to analyse the ecological profiles of the Ny. intermedia and Ny. neivai populations in a sympatric area in the Brazilian savannah along the banks of the Velhas River. Captures were performed from July 2003-June 2005 in two distinct environments: a gallery forest with various degrees of anthropogenic modification and animal shelters. A total of 20,508 Ny. neivai (86%) and Ny. intermedia (14%) sandflies were collected. The difference between the proportions of the sandflies that were collected (Ny. neivai/Ny. intermedia) per bank was significant. The right bank presented a greater number of sandflies (65%) and more preserved vegetation. The abundance of Ny. neivai was higher than that of Ny. intermedia on both banks. The results demonstrate that anthropic activities can affect the sandfly populations in this area, thereby leading to a reduction in species abundance. Nevertheless, the environments with higher levels of antropogenic modification displayed sandfly population numbers that favour the Leishmania transmission cycle.

Biogeographical aspects of the occurrence of Nyssomyia neivai and Nyssomyia intermedia (Diptera: Psychodidae) in a sympatric area of the Brazilian savannah

Nyssomyia intermedia and Nyssomyia neivai constitute a species complex associated with Leishmania transmission. The aim of this study was to analyse the ecological profiles of the Ny. intermedia and Ny. neivai populations in a sympatric area in the Brazilian savannah along the banks of the Velhas River. Captures were performed from July 2003-June 2005 in two distinct environments: a gallery forest with various degrees of anthropogenic modification and animal shelters. A total of 20,508 Ny. neivai (86%) and Ny. intermedia (14%) sandflies were collected. The difference between the proportions of the sandflies that were collected (Ny. neivai/Ny. intermedia) per bank was significant. The right bank presented a greater number of sandflies (65%) and more preserved vegetation. The abundance of Ny. neivai was higher than that of Ny. intermedia on both banks. The results demonstrate that anthropic activities can affect the sandfly populations in this area, thereby leading to a reduction in species abundance. Nevertheless, the environments with higher levels of antropogenic modification displayed sandfly population numbers that favour the Leishmania transmission cycle.

Natural Leishmania sp. reservoirs and phlebotomine sandfly food source identification in Ibitipoca State Park, Minas Gerais, Brazil.

Leishmania spp are distributed throughout the world and different species are associated with varying degrees of disease severity. However, leishmaniasis is thought to be confined to areas of the world where its insect vectors, sandflies, are present. Phlebotomine sandflies obtain blood meals from a variety of wild and domestic animals and sometimes from humans. These vectors transmit Leishmania spp, the aetiological agent of leishmaniasis. Identification of sandfly blood meals has generally been performed using serological methods, although a few studies have used molecular procedures in artificially fed insects. In this study, cytochrome b gene (cytB) polymerase chain reaction (PCR) was performed in DNA samples isolated from 38 engorged Psychodopygus lloydi and the expected 359 bp fragment was identified from all of the samples. The amplified product was digested using restriction enzymes and analysed for restriction fragment length polymorphisms (RFLPs). We identified food sources for 23 females; 34.8% yielded a primate-specific banding profile and 26.1% and 39.1% showed banding patterns specific to birds or mixed restriction profiles (rodent/marsupial, human/bird, rodent/marsupial/human), respectively. The food sources of 15 flies could not be identified. Two female P. lloydi were determined to be infected by Leishmania using internal transcribed spacer 1 and heat shock protein 70 kDa PCR-RFLP. The two female sandflies, both of which fed on rodents/marsupials, were further characterised as infected with Leishmania (Viannia) braziliensis. These results constitute an important step towards applying methodologies based on cytB amplification as a tool for identifying the food sources of female sandflies.

Natural Leishmania sp. reservoirs and phlebotomine sandfly food source identification in Ibitipoca State Park, Minas Gerais, Brazil

Leishmania spp are distributed throughout the world and different species are associated with varying degrees of disease severity. However, leishmaniasis is thought to be confined to areas of the world where its insect vectors, sandflies, are present. Phlebotomine sandflies obtain blood meals from a variety of wild and domestic animals and sometimes from humans. These vectors transmit Leishmania spp, the aetiological agent of leishmaniasis. Identification of sandfly blood meals has generally been performed using serological methods, although a few studies have used molecular procedures in artificially fed insects. In this study, cytochrome b gene (cytB) polymerase chain reaction (PCR) was performed in DNA samples isolated from 38 engorged Psychodopygus lloydi and the expected 359 bp fragment was identified from all of the samples. The amplified product was digested using restriction enzymes and analysed for restriction fragment length polymorphisms (RFLPs). We identified food sources for 23 females; 34.8% yielded a primate-specific banding profile and 26.1% and 39.1% showed banding patterns specific to birds or mixed restriction profiles (rodent/marsupial, human/bird, rodent/marsupial/human), respectively. The food sources of 15 flies could not be identified. Two female P. lloydi were determined to be infected by Leishmania using internal transcribed spacer 1 and heat shock protein 70 kDa PCR-RFLP. The two female sandflies, both of which fed on rodents/marsupials, were further characterised as infected with Leishmania (Viannia) braziliensis. These results constitute an important step towards applying methodologies based on cytB amplification as a tool for identifying the food sources of female sandflies.

Lutzomyia diamantinensis sp. nov., a new phlebotomine species (Diptera: Psychodidae) from a quartzite cave in Diamantina, state of Minas Gerais, Brazil.

A new species of Brazilian phlebotomine sandfly found in Brazil, municipality of Diamantina, state of Minas Gerais, is described based on males and females collected in a quartzite cave. The body of spermathecae is continuous to the individual duct, lanky and tapering at the end, with conical shaped, not striated and presenting the head with dense setae. The male presents gonostyle with four spines and a small subterminal seta and gonocoxite with one group of persistent setae. The paramere is simple with a group of small setae on the dorsal apex. The morphological features of this new species permit its inclusion in the migonei group.

Hourly activity and natural infection of sandflies (Diptera: psychodidae) captured from the aphotic zone of a cave, minas gerais state, Brazil.

Sandflies are holometabolous insects that are of great epidemiological importance in the neotropical region as vectors of leishmaniases. Caves are ecotopes that significantly differ from external environments and, among the insects that live or visit their internal area and adjacent environment, sandflies are commonly found. Based on this context, the objective of this work was to examine the period of activity of sandflies in the cave environment in the aphotic zone. Thus, four sandfly captures were conducted, one in each season of the year, in a cave where studies on the bioecological aspects of sandfly fauna have been conducted since 2008. In this same study, we have also noticed the presence of flagellates in some captured females. Catches were carried out for 24 hours using a Shannon trap, light bait, and cave walls were actively searched. We collected a total of 638 sandflies, representing 11 species. The most abundant species and with more intense period of activity were, in descending order: Lu. cavernicola (62%), Ev. spelunca (16%) and Ev. sallesi (14%). A total of 69 females were dissected to check for natural infection, and in five specimens we found living flagellated forms: two Ev. spelunca, two Ev. sallesi and one Sc. sordellii. This study shows that the activity of some species caught in the aphotic zone of the cave, especially Lu. cavernicola, differs from what has already been reported in previous sandfly captures, which are almost always conducted at night and during twilight. The existence of sandflies that were naturally infected with flagellates and the lack of awareness regarding the behaviour of sandflies in cave environments are strong indicators of the need for further study on this group of insects in this ecotope, as a safety measure to protect the visitors of such environment.

Diversity of phlebotomine sand flies (Diptera: Psychodidae) in Ibitipoca State Park, Minas Gerais, Brazil.

Leishmaniasis is a complex of zoonotic diseases that are endemic to many Brazilian states. They are transmitted to the vertebrates by the bite of the hematophagous female sand fly (Diptera: Psychodidae) vectors. Despite the increasing occurrence of visceral and cutaneous leishmaniasis cases in large urban centers, their transmission continues to occur primarily in a wild environment and may be associated with professional activities, ecotourism activities, or both. This study investigates the ecological parameters of the sand flies present in Ibitipoca State Park, Minas Gerais, Brazil. During 2009, systematic collections of sand flies were made monthly using HP light traps installed at five sites, including three natural settings (a cave, riparian vegetation, and a rain forest), the tourist and researchers' accommodations, and a surrounding domestic livestock area. In total, 161 sand flies (seven species) were collected, the most abundant, particularly in the surrounding domestic livestock area, being Lutzomyia (Psychodopygus) lloydi (Antunes, 1937). Furthermore, a previously unidentified Lutzomyia (Sciopemyia) sp. was prevalent in the cave environment. There are no existing records of the occurrence of leishmaniasis in Ibitipoca State Park; however, the some species of the subgenus Psychodopygus are known vectors of Leishmania spp in Brazil. Hence, the presence of a species of this genus in areas surrounding the park may represent a risk to ecotourism and the local inhabitants. Our study shows the importance of regular monitoring of the various areas used by humans to determine the distribution and spread of sand fly vectors for preventive management to forestall potential risk to health and consequent effect on ecotourists.

Description of a new phlebotomine species, Martinsmyia reginae sp. nov. (Diptera: Psychodidae: Phlebotominae) from a cave in the state of Tocantins Brazil.

As inhabitants of forested areas, caves and anthropic environments, the phlebotomines deserve special attention because some species are able to transmit trypanosomatids, bacteria and viruses to vertebrates. Phlebotomines are also a nuisance because they cause painful bites, which may ultimately produce allergic manifestations. The lack of information about the presence and behaviours of sand flies in caves has aroused the curiosity of researchers for a long time. In the present paper, we describe a new species of sand fly that was captured in a cave located in the municipal district of Arraias in the southeastern region of the state of Tocantins. The morphological features of this new species permit it to be included in the alphabetica group of the Martinsmyia genus.

Description of a new phlebotomine species, Martinsmyia reginae sp. nov. (Diptera: Psychodidae: Phlebotominae) from a cave in the state of Tocantins Brazil

As inhabitants of forested areas, caves and anthropic environments, the phlebotomines deserve special attention because some species are able to transmit trypanosomatids, bacteria and viruses to vertebrates. Phlebotomines are also a nuisance because they cause painful bites, which may ultimately produce allergic manifestations. The lack of information about the presence and behaviours of sand flies in caves has aroused the curiosity of researchers for a long time. In the present paper, we describe a new species of sand fly that was captured in a cave located in the municipal district of Arraias in the southeastern region of the state of Tocantins. The morphological features of this new species permit it to be included in the alphabetica group of the Martinsmyia genus.

Description of Evandromyia grimaldii sp. nov. (Diptera: Psychodidae), a new phlebotomine species from the state of Espírito Santo, Southeast Brazil.

Phlebotomine sand flies present great species diversity, especially in Brazil, where there are about 300 known species. This work describes a new species of sand fly found in the Brazilian state of Espírito Santo, in the Reserva Biológica de Duas Bocas, municipality of Cariacica. Spermathecae with superficial striations and the common duct longer than the genital fork permit the inclusion of the new species in the subgenus Evandromyia s. str., series rupicola. The new species resemble Ev. rupicola from which it may be distinguished by the aspects of the spermatheca and the length of the genital filament of the male, longer in the new species.