Remarkable diversity, new records and Leishmania detection in the sand fly fauna of an area of high endemicity for cutaneous leishmaniasis in Acre state, Brazilian Amazonian Forest.

The species richness of Amazonian phlebotomines is considered to be one of the highest in the world. In the present study, we investigated the richness and diversity of phlebotomine fauna in Xapuri city, Acre state, Western Brazilian Amazonia, which is an area that is highly endemic for cutaneous leishmaniasis. Sand fly collections were performed monthly from August 2013 to July 2015 (288 h total of sampling effort) in intradomiciliary, peridomiciliary, and forested environments of two localities. Collected females were dissected, microscopically examined for flagellates in their guts, and preserved in ethanol. A total of 21,197 specimens comprising 14 genera and 57 species were collected, and the majority of these were Nyssomyia, Psychodopygus, and Trichophoromyia genera. Three new records of phlebotomine species for Acre are presented here, including Brumptomyia brumpti, Psathyromyia pradobarrientosi, and for the first time in Brazil, Th. omagua. In Xapuri, the phlebotomine fauna of different ecotopes was varied in regard to abundance, diversity, and frequency, and they included proven and permissive vectors of Leishmania spp. The fauna discovered in the forested areas (57 species) was richer and more diverse than was that (33 species) identified in the peri­ and intra-domiciles. The identification of Leishmania subgenera that were present in sand fly guts according to SSU rRNA sequences revealed ten and three species harboring Leishmania of subgenera Viannia and Leishmania (most likely Leishmania amazonensis), respectively. The presence of Leishmania (Leishmania) in sand flies are reported here for the first time in Acre. The presence of L. (Viannia) spp. in Brumptomyia sp. and Lutzomyia sherlocki. and the occurrence of mixed infections with Leishmania of both subgenera in Ps. lainsoni have been reported for the first time in Brazil. Taken together, data from previous studies and from the present study highlight the remarkable complexity of phlebotomine fauna that is possibly due to the well-preserved Xapuri forested areas sustaining vital economic activities of plant extraction and ecological tourism. Our findings also provide new insights into the ongoing adaptation of Trichophoromyia and Psychodopygus species to human habitats.

Culicidae evolutionary history focusing on the Culicinae subfamily based on mitochondrial phylogenomics.

Mosquitoes are insects of medical importance due their role as vectors of different pathogens to humans. There is a lack of information about the evolutionary history and phylogenetic positioning of the majority of mosquito species. Here we characterized the mitogenomes of mosquito species through low-coverage whole genome sequencing and data mining. A total of 37 draft mitogenomes of different species were assembled from which 16 are newly-sequenced species. We datamined additional 49 mosquito mitogenomes, and together with our 37 mitogenomes, we reconstructed the evolutionary history of 86 species including representatives from 15 genera and 7 tribes. Our results showed that most of the species clustered in clades with other members of their own genus with exception of Aedes genus which was paraphyletic. We confirmed the monophyletic status of the Mansoniini tribe including both Coquillettidia and Mansonia genus. The Aedeomyiini and Uranotaeniini were consistently recovered as basal to other tribes in the subfamily Culicinae, although the exact relationships among these tribes differed between analyses. These results demonstrate that low-coverage sequencing is effective to recover mitogenomes, establish phylogenetic knowledge and hence generate basic fundamental information that will help in the understanding of the role of these species as pathogen vectors.

Phlebotomine (Diptera: Psychodidae) fauna in a cavern containing cave paintings and its surrounding environment, Central-West Brazil.

The interior of caves usually presents climate, light, soil, flora and fauna that distinguish it from the external environment. Among the sandfly species found in caves, several may act as vectors of Leishmania spp. and others are a nuisance to humans due to their painful bites. This study sought to identify the phlebotomine fauna in a cavern with cave paintings ("Gruta do Pitoco") which attract tourists in Alcinopólis municipality, Mato Grosso do Sul state, Brazil. The sandflies were collected from March 2014 to September 2015, with automatic light traps, installed monthly, in the entrance and interior of the cave and outside it. A total of 7645 specimens belonging to 27 species were collected. Lutzomyia dispar, Martinsmyia oliverai and those of the Lu. longipalpis complex (Lu. cruzi and Lu. longipalpis) were the predominant species. Among the three environments sampled, no great difference between species richness was observed. Comparing these environments, the interior of the cave presented the lowest diversity index due to the high frequency of Lu. dispar (46%). For the first time, specimens (only females) of the genus Deanemyia were collected in Mato Grosso do Sul state. Moreover, the study revealed the sympatric occurrence of Lu. cruzi and Lu. longipalpis, both proven vectors of L. infantum, and also Lu. dispar, a highly anthropophilic species, which often causes annoyance to those who visit the cave.

Attractiveness of black and white modified Shannon traps to phlebotomine sandflies (Diptera, Psychodidae) in the Brazilian Amazon Basin, an area of intense transmission of American cutaneous leishmaniasis.

In the Amazon region the phlebotomine fauna is considered one of the most diverse in the world. The use of Shannon traps may provide information on the anthropophily of the species and improve the traps' performance in terms of diversity and quantity of insects collected when white and black colored traps are used together. This study sought to verify the attractiveness of the traps to the phlebotomine species of the Brazilian Amazon basin using Shannon traps under these conditions. The insects were collected using two Shannon traps installed side by side, one white and the other black, in a primary forest area of the municipality of Xapuri, Acre, Brazil. Samples were collected once a month during the period August 2013 to July 2015. A sample of females was dissected to test for natural infection by flagellates. A total of 6,309 (864 males and 5,445 females) specimens (36 species) were collected. Psychodopygus carrerai carrerai (42%), Nyssomyia shawi (36%), and Psychodopygus davisi (13%), together represented 90% of the insects collected. Nyssomyia shawi and Psychodopygus davisi were more attracted by the white color. Specimens of Nyssomyia shawi, Nyssomyia whitmani, and Psychodopygus hirsutus hirsutus were found naturally infected by flagellates in the mid and hindgut. This is the first study in Acre state using and comparing both black and white Shannon traps, demonstrating the richness, diversity, and anthropophilic behavior of the phlebotomine species and identifying proven and putative vectors of the etiological agents of leishmaniasis.

Vector aspects in risk areas for sylvatic yellow fever in the state of Mato Grosso do Sul, Brazil

Epizootics and yellow fever epidemics in 2008 in Brazil, Paraguay and Argentina marked a significant progression of the disease further south in the continent affecting non-vaccinated human populations. The state of Mato Grosso do Sul is located between the tropical endemic region and the subtropical epidemic region and, despite being an area recommended for vaccination since the 1980s, human and animal cases of the disease are still registered. The purpose of this study was to present the results of entomological surveys of probable sites of infection (PSI) geographically describing these areas regarding human cases and disease suspicion reported in 2008. Thirteen locations in nine municipalities were investigated. A total of 305 females of the genus Haemagogus and Sabethes and five specimens of Aedes albopictus were obtained. The genus Haemagogus was more abundant but Sabethes presented twice as many species (3 x 6) and higher distribution within the collection points. The most abundant species was Hg. janthinomys, with 102 individuals in one location. The other species found were: Hg. leucocelaenus, Hg. spegazzinii, Sa. albiprivus, Sa. belisarioi, Sa. chloropterus, Sa. glaucodaemun, Sa. intermedius and Sa. soperi. It was not possible to identify predominant species in relation to environmental and geographical characteristics of the collection points. Of the nine human cases, eight corresponded to persons not resident in the State, seven of them tourists. MS features a large diversity of rural and wildlife tourist attractions where the yellow fever vectors described in this work are present. In this sense, the monitoring of vaccination coverage and flow of non-vaccinated people through the area are crucial to controlling the disease.

Detection of Culex flavivirus and Aedes flavivirus nucleotide sequences in mosquitoes from parks in the city of São Paulo, Brazil.

The dengue viruses are widespread in Brazil and are a major public health concern. Other flaviviruses also cause diseases in humans, although on a smaller scale. The city of São Paulo is in a highly urbanized area with few green spaces apart from its parks, which are used for recreation and where potential vertebrate hosts and mosquito vectors of pathogenic Flavivirus species can be found. Although this scenario can contribute to the transmission of Flavivirus to humans, little is known about the circulation of members of this genus in these areas. In light of this, the present study sought to identify Flavivirus infection in mosquitoes (Diptera: Culicidae) collected in parks in the city of São Paulo. Seven parks in different sectors of the city were selected. Monthly mosquito collections were carried out in each park from March 2011 to February 2012 using aspiration and traps (Shannon and CD C-CO2). Nucleic acids were extracted from the mosquitoes collected and used for reverse-transcriptase and real-time polymerase chain reactions with genus-specific primers targeting a 200-nucleotide region in the Flavivirus NS5 gene. Positive samples were sequenced, and phylogenetic analyses were performed. Culex and Aedes were the most frequent genera of Culicidae collected. Culex flavivirus (CxFV)-related and Aedes flavivirus (AEFV)- related nucleotide sequences were detected in 17 pools of Culex and two pools of Aedes mosquitoes, respectively, among the 818 pools of non-engorged females analyzed. To the best of our knowledge, this is the first report of CxFV and AEFV in the city of São Paulo and Latin America, respectively. Both viruses are insect- specific flaviviruses, a group known to replicate only in mosquito cells and induce a cytopathic effect in some situations. Hence, our data suggests that CxFV and AEFV are present in Culex and Aedes mosquitoes, respectively, in parks in the city of São Paulo. Even though Flavivirus species of medical importance were not detected, surveillance is recommended in the study areas because of the presence of vertebrates and mosquitoes that could act as amplifying hosts and vectors of flaviviruses, providing the required conditions for circulation of these viruses.

Feeding habits of mosquitoes (Diptera: Culicidae) in an area of sylvatic transmission of yellow fever in the state of São Paulo, Brazil.

BACKGROUND: The reintroduction of sylvatic yellow fever in the state of São Paulo after about six decades was confirmed in the Northwestern region in 2000, where in 2008 there also occurred an important epizootic. The purpose of this study was to investigate the feeding habits of culicids potentially involved in the sylvatic transmission of the virus in this region. METHODS: Specimens were collected in 24 forested localities at ground level with hand nets and mouth aspirators. Collections were made quarterly between October 2006 and July 2008 during daylight hours. Blood-meal identification was carried out in mosquitoes of the tribes Aedini, Mansoniini and Sabethini. The biotin/avidin sandwich ELISA was employed to determine six source types: bird, bovine, equine, rat, human and monkey. RESULTS: A total of 24,879 females of the three tribes were obtained, 245 (0.98%) of which were engorged. The presence of three different blood sources per engorged female was the predominant situation, and included 35.10% of the total of samples processed. Samples with two or four different sources were represented by 25.31% and 25.71%, of the specimens, respectively, while just 9.39% had only one type and 1.22%, five different sources. Aedes scapularis, Ae. serratus (Group), Psorophora albigenu and Ps. ferox were the most abundant species and accounted for about 95% of the engorged specimens. Of the principal vector species, Haemagogus janthinomys/capricornii was found with bird, bovine and primate blood. These sources were predominant and alternated top ranking as the most frequent source according to the mosquito species and collection site. In general, primate blood was the most prevalent source. CONCLUSIONS: The human population of the region visits this ecotone frequently, which indicates the need for the periodical assessment of vaccination coverage against yellow fever. The frequency of non-human primate blood source in mosquito species that show minor vector importance in yellow fever virus transmission deserves attention. The eclectic feeding habits and some aspects of the interactions between potential vectors and reservoirs of yellow fever may be associated with the habitat fragmentation characteristic of the region. We recommend that further studies on the capacity and vector competence be performed on secondary vectors in extra-Amazonian region.

Mosquito fauna in municipal parks of São Paulo City, Brazil: a preliminary survey.

A mosquito faunal survey was conducted from October 2010 to February 2011 in the municipal parks of São Paulo City, Brazil. A total of 7,015 specimens of 53 taxonomic categories grouped into 12 genera (Aedes, Anopheles, Coquilletidia, Culex, Limatus, Lutzia, Mansonia, Psorophora, Toxorhynchites, Trichoprosopon, Uranotaenia, and Wyeomyia) were collected. The largest and most peripheral parks showed greater species richness compared to smaller and more centralized parks.

Mosquitoes in degraded and preserved areas of the Atlantic Forest and potential for vector-borne disease risk in the municipality of São Paulo, Brazil.

In order to assess the epidemiological potential of the Culicidae species in remaining areas of the Brazilian Atlantic Forest, specimens of this family were collected in wild and anthropic environments. A total of 9,403 adult mosquitoes was collected from May, 2009 to June, 2010. The most prevalent among species collected in the wild environment were Anopheles (Kerteszia) cruzii, the Melanoconion section of Culex (Melanoconion), and Aedes serratus, while the most common in the anthropic site were Coquillettidia chrysonotum/albifera, Culex (Culex) Coronator group, and An. (Ker.) cruzii. Mosquito richness was similar between environments, although the abundance of individuals from different species varied. When comparing diversity patterns between environments, anthropic sites exhibited higher richness and evenness, suggesting that environmental stress increased the number of favorable niches for culicids, promoting diversity. Increased abundance of opportunistic species in the anthropic environment enhances contact with culicids that transmit vector-borne diseases.

Effects of Artificial Flooding for Hydroelectric Development on the Population of Mansonia humeralis (Diptera: Culicidae) in the Paraná River, São Paulo, Brazil.

The closure of two phases of the dam at the Porto Primavera Hydroelectric Plant on the Paraná River flooded a flawed system located in the Municipality of Presidente Epitácio, São Paulo state, favoring the proliferation of aquatic weeds. This study aimed to observe the population of Mansonia humeralis in the area, monitoring the richness, diversity, and dominance of this species both before and during different phases of reservoir flooding as well as evaluate its possible consequences concerning human and animal contact. Adult mosquitoes were collected monthly in the following periods: at the original level, after the first flood, and after the maximum level had been reached between 1997 and 2002. Collection methods used were an aspirator, a Shannon trap, and the Human Attractive Technique. A total of 30,723 mosquitoes were collected, Ma. humeralis accounting for 3.1% in the preflood phase, 59.6% in the intermediate, and 53.8% at maximum level. This species is relevant to public health, since the prospect of continued contact between Ma. humeralis and the human population enhances the dam's importance in the production of nuisance mosquitoes, possibly facilitating the transmission of arboviruses. Local authorities should continue to monitor culicid activity through sustainable entomological surveillance.

Ecological aspects of mosquitoes (Diptera: Culicidae) in an Atlantic forest area on the north coast of Rio Grande do Sul State, Brazil.

Mosquito diversity was determined in an area located on the southern limit of the Atlantic Forest on the north coast of Rio Grande of Sul State. Our major objective was to verify the composition, diversity, and temporal distribution of the mosquito fauna, and the influence of temperature and rainfall. Samplings were performed monthly between December, 2006 and December, 2008, in three biotopes: forest, urban area, and transition area, using CDC light traps and a Nasci vacuum. A total of 2,376 specimens was collected, from which 1,766 (74.32%) were identified as 55 different species belonging to ten genera. Culex lygrus, Aedes serratus, and Aedes nubilus were dominant (eudominant) and constant throughout samplings. The forest environment presented the highest species dominance (D(S) =0.20), while the transition area showed the highest values of diversity (H'=2.55) and evenness (J'=0.85). These two environments were the most similar, according to the Morisita-Horn Index (I(M-H) =0.35). Bootstrap estimates showed that 87.3% of the species occurring in the region were detected. The seasonal pattern showed a greater abundance of mosquitoes between May and October, indicating the period to intensify entomological surveillance in that area.

[Richness and abundance of Culicidae (Diptera) in an area impacted, Mato Grosso do Sul, Midwestern Brazil]. / Riqueza e abundância de Culicidae (Diptera) em área impactada, Mato Grosso do Sul, Brasil.

The damming of Paraná River for the construction of Porto Primavera Hydroelectric Power Plant, between the states of Mato Grosso do Sul, Midwestern Brazil and São Paulo, Southeastern Brazil, has changed the ecological relationships in the area. The objective of the study was to describe the fauna that can be potential vectors in this reservoir, 2 km away from its right bank, in Bataguassu, Mato Grosso do Sul, before flooding. Mosquitoes were monthly captured in different environments from July 1997 to November 1999. During this period 16,553 adult specimens and 1,795 immature forms were collected with richness of 86 different species of adults and 44 different species of immature forms. The fauna presented a richness that included species of epidemiological importance, as Anopheles darlingi and other mosquitoes with potential for arbovirus transmission.