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INTRODUÇÃO: O objetivo deste estudo foi mensurar a diversidade de espécies de culicídeos, descrever sua abundância e variação sazonal em áreas urbanas e matas de São José do Rio Preto, SP, e discutir o risco de ocorrência de arboviroses. MÉTODOS: Coletas de larvas e de mosquitos adultos foram realizadas mensalmente, em 2006 e 2007, em área urbana e em quatro fragmentos de mata. No perímetro urbano, coletaram-se larvas nos sítios mais prováveis de oviposição para mosquitos do gênero Culex e nas matas foi realizada a coleta de mosquitos adultos, sendo que em duas utilizaram-se armadilhas CDC à noite e, em duas, aspirador de Nasci de dia. RESULTADOS: Na área urbana identificaram-se 34 espécies de culicídeos em um total de 8.683 exemplares; destes, 80,7% corresponderam ao Culex quinquefasciatus, 9,6% ao Culex grupo Coronator, 3,2% ao Aedes albopictus (3,2%) e 1,1% ao Ochlerotatus fluviatilis. A abundância de larvas de Cx. quinquefasciatus correlacionou-se negativamente com a chuva. Nas quatro matas, foram coletados 2.268 mosquitos distribuídos entre 10 gêneros, 46 espécies ou grupos. As mais abundantes foram Aedeomyia squamipennis, Culex. coronator, Culex (Mel.) seção Melanoconion, Culex declarator, Ochlerotatus scapularis, Anopheles triannulatus, Culex bidens/interfor e Culex habilitator/pseudojhantinosoma. CONCLUSÕES: A abundância de Cx. quinquefasciatus na área urbana e a presença de outros culicídeos nas áreas urbanas e de matas apontam para a possibilidade de transmissão do vírus do Nilo Ocidental e de outras arboviroses em São José do Rio Preto e outras cidades do Brasil, sendo fundamental o estabelecimento de medidas visando à vigilância destas arboviroses. INTRODUCTION: The objectives for this study were to measure the diversity of Culicidae species, describe their abundance and seasonal variation in São José do Rio Preto, SP, and discuss the risk of arbovirus infections. METHODS: The collection of larval and adult mosquitoes was conducted monthly from 2006 to 2007 in an urban area and four sections of forested land. In the urban area, larvae were collected from sites where oviposition by Culex mosquitoes was most likely to occur. At two of the four sites in the forested land, adult mosquitoes were collected with the use of CDC traps at night, and a Nasci aspirator was used in the daytime at the two other collection sites. RESULTS: In the urban area, 34 Culicidae species were identified out of a total sample of 8,683 specimens; of these specimens, 80.7% were Culex quinquefasciatus, 9.6% were Culex coronator, 3.2% were Aedes albopictus, and 1.1% were Ochlerotatus fluviatilis. The abundance of Cx. quinquefasciatus larvae was negatively related to rainfal. In the woods, 2,268 mosquitoes were collected, representing 10 genera and 46 species. The most abundant mosquito species were Aedeomyia squamipennis, Culex coronator, Culex (Mel.) Melanoconion section, Culex declarator, Ochlerotatus scapularis, Anopheles triannulatus, Culex bidens/interfor and Culex habilitator/pseudojhantinosoma. CONCLUSIONS: The abundance of Cx. quinquefasciatus in the urban area and the presence of other Culicidae species in urban areas and forested land point to the possibility of the transmission of West Nile virus and other arbovirus infections in São José do Rio Preto and other cities. Thus, the enacting of measures aimed at the surveillance of these arbovirus infections is essential.

Culex is the largest subgenus within the genus Culex that includes important vectors of diseases. The correct identification of mosquitoes is critical for effective control strategies. Wing geometric morphometrics (WGM) has been used to identify mosquito species alongside traditional identification methods. Here, WGM was used for eleven Culex species from São Paulo, Brazil, and one from Esquel, Argentina. Adult mosquitoes were collected using CDC (Centers for Disease Control) traps, morphologically identified and analyzed by WGM. The canonical variate analysis (CVA) was performed and a Neighbor-joining (NJ) tree was constructed to illustrate the patterns of species segregation. A cross-validated reclassification test was also carried out. From 110 comparisons in the cross-validated reclassification test, 87 yielded values higher than 70%, with 13 comparisons yielding 100% reclassification scores. Culexquinquefasciatus yielded the highest reclassification scores among the analyzed species, corroborating with the results obtained by the CVA, in which Cx. quinquefasciatus was the most distinct species. The high values obtained at the cross-validated reclassification test and in the NJ analysis as well as the segregation observed at the CVA made it possible to distinguish among Culex species with high degrees of confidence, suggesting that WGM is a reliable tool to identify Culex species of the subgenus Culex.

Yellow fever virus (YFV) is an endemic arbovirus in parts of Africa and the Americas. In Brazil, following the eradication of the urban transmission cycle, YFV is maintained in a sylvatic cycle involving several species of neotropical primates and mosquitoes of the genera Haemagogus and Sabethes, which serve as primary and secondary vectors, respectively. During the 2016–2019 outbreak in São Paulo State, a total of 3731 mosquito pools were collected from sites with ongoing epizootic events in 192 municipalities. The RT-qPCR analysis detected YFV in 46 pools (1.4%) across nine mosquito species, including both primary and secondary vectors, as well as species from the genera Aedes and Psorophora. Differences in viral loads were observed among species. While Aedes aegypti was not found to be positive, the detection of natural YFV infection in other Aedes species raises concerns about potential virus reurbanization. Further studies are needed to clarify the role of additional mosquito species in YFV transmission in Brazil.

Brazil constitutes a hotspot for arbovirus emergence, posing persistent public health challenges. In São José do Rio Preto (SJdRP), dengue has historically predominated, whereas only 62 chikungunya virus (CHIKV) cases were reported between 2015 and 2023, suggesting cryptic circulation. However, from 2024 onwards, CHIKV notifications began to increase across São Paulo State. Because arboviral febrile illnesses frequently overlap clinically, early detection necessitates robust surveillance systems. To address this, mosquito collections were conducted over 15 months in SJdRP and screened for CHIKV RNA. Positive specimens were further analyzed through virus isolation and whole-genome sequencing. Overall, 5.6% of specimens tested positive, with prevalence rising from December 2023 and peaking in May 2024, coinciding with human cases. Phylogenetic reconstruction indicated multiple introductions, primarily from southeastern Brazil, driving viral establishment. These events likely facilitated transmission within a dense, susceptible vector population, contributing to extensive spread and culminating in a major CHIKV outbreak in northwestern São Paulo.

Zoological gardens in forest areas host a large diversity of vertebrate species (exotic and indigenous, free-living and captive, migrant and resident), resulting in an artificial proximity of animal species that would never share the same environment in natural conditions. The presence of mosquitoes enables the transmission of vector-borne pathogens, as is the case with avian malaria parasites. The mild infections in some bird species may become a serious threat to others that do not possess a natural resistance. Thus, the identification of the potential vectors of these parasites is important for future control of these agents, aiming at the bird species conservation. In this study, we collected over 2000 mosquitoes in Sao Paulo Zoo and analyzed them through molecular methods. Six different mosquito species yielded positive for the targeted parasite DNA. We demonstrated that these culicids had fed mainly on bird species and we reported three mosquito species that have never been previously incriminated as potential vectors of these parasites, enabling the use of more specific measures for vigilance and mosquito control. Avian malaria parasites are widespread parasites transmitted by Culicidae insects belonging to different genera. Even though several studies have been conducted recently, there is still a lack of information about potential vectors of Plasmodium parasites, especially in Neotropical regions. Former studies with free-living and captive animals in Sao Paulo Zoo showed the presence of several Plasmodium and Haemoproteus species. In 2015, a pilot study was conducted at the zoo to collect mosquitoes in order to find out (i) which species of Culicidae are present in the study area, (ii) what are their blood meal sources, and (iii) to which Plasmodium species might they be potential vectors. Mosquitoes were morphologically and molecularly identified. Blood meal source and haemosporidian DNA were identified using molecular protocols. A total of 25 Culicidae species were identified, and 6 of them were positive for Plasmodium/Haemoproteus DNA. Ten mosquito species had their source of blood meal identified, which were mainly birds, including some species that were positive for haemosporidian parasites in the former study mentioned. This study allowed us to expand the list of potential vectors of avian malaria parasites and to improve our knowledge of the evolutionary and ecological relationships between the highly diverse communities of birds, parasites, and vectors present at Sao Paulo Zoo.

The identification of mosquito species is necessary for determining the entomological components of disease transmission. However, identification can be difficult in species that are morphologically similar. The cytochrome c oxidase subunit I (COI) DNA barcode region is considered a valuable and reliable diagnostic tool for mosquito species recognition, including those that belong to species complexes. Mansonia mosquitoes are found in forests near swampy areas. They are nocturnal and are highly attracted to light. Hematophagous adult females exhibit aggressive biting behavior and can become infected with and transmit pathogens during their feeding, including some epizootic viruses and avian malaria. In Brazil, twelve Mansonia species have been reported. In a recent study from the São Paulo Zoo in Brazil, three morphologically distinct species were collected and identified, namely: Mansonia (Mansonia) indubitans, Ma. (Man.) pseudotitillans and Ma. (Man.) titillans. However, confirmation of these species by molecular identification was unsuccessful due to a lack of COI sequences in the GenBank database. Thus, this research aimed to describe the COI DNA barcode sequences of some morphologically characterized Mansonia (Man.) species from Brazil and to determine their utility in delimiting species collected from the Atlantic Forest and Brazilian Savanna. Accordingly, we provide tools for the genetic identification of species that play a significant role in pathogen transmission in wildlife and potentially humans. We show that the delimitation of Mansonia species via five different approaches based on COI DNA sequences (BI, NJ, ASAP, bPTP and GMYC) yield basically the same groups identified by traditional taxonomy, and we provide the identification of specimens that were previously identified only up to the subgenus level. We also provide COI sequences from two Mansonia species that were not previously available in sequence databases, Ma. wilsoni and Ma. pseudotitillans, and thus contribute to the ongoing global effort to standardize DNA barcoding as a molecular means of species identification.

Yellow Fever (YF) is a severe disease caused by Yellow Fever Virus (YFV), endemic in some parts of Africa and America. In Brazil, YFV is maintained by a sylvatic transmission cycle involving non-human primates (NHP) and forest canopy-dwelling mosquitoes, mainly Haemagogus -spp and Sabethes- spp. Beginning in 2016, Brazil faced one of the largest Yellow Fever (YF) outbreaks in recent decades, mainly in the southeastern region. In São Paulo city, YFV was detected in October 2017 in Aloutta monkeys in an Atlantic Forest area. From 542 NHP, a total of 162 NHP were YFV positive by RT-qPCR and/or immunohistochemistry, being 22 Callithrix- spp. most from urban areas. Entomological collections executed did not detect the presence of strictly sylvatic mosquitoes. Three mosquito pools were positive for YFV, 2 Haemagogus leucocelaenus , and 1 Aedes scapularis . In summary, YFV in the São Paulo urban area was detected mainly in resident marmosets, and synanthropic mosquitoes were likely involved in viral transmission.

Sylvatic yellow fever (SYF) was recently a health issue in Brazil (2016-2019) because transmission was facilitated by a high density of vectors, amplifying hosts, and low vaccine coverage of the human population, especially in urban forests in the Southeast Region of Brazil. Moreover, urban forest edges are more likely to have contact between human and sylvatic vector mosquito populations. Here, we show the association between abiotic and biotic features of tree holes as 'Haemagogus leucocelaenus' rearing sites in Cantareira State Park in Atlantic Forest edges. The analyzed physical features of the tree holes were diameter at breast height, tree hole opening diameter, depth, trunk diameter, tree hole volume, collected volume, height (varying from 0.02 to 4.2 m above ground), and the presence of Culicidae species other than 'Hg. leucocelaenus'. We analyzed 105 positive and 68 negative water samples for larval presence and found no differences between them, suggesting the lack of specific physical characteristics in these categories. 'Hg. leucocelaenus' larval abundance was correlated with the collected volume and opening diameter of tree holes. The tree species that most represented negative breeding sites were 'Euplassa cantareirae', 'Guarea macrophylla', 'Psychotria suterella', and 'Tibouchina pulchra'. Four significant clusters as areas with a high risk of SYV were identified by Get-Ordis spatial analysis. Although 'Hg. leucocelaenus' larvae were found in tree holes with high water levels, their occurrence was regulated by that of other mosquito species. Our findings contribute to clarifying immature vector ecology in tree holes related to human exposure to SYF in urban forest edges.

The objectives for this study were to measure the diversity of Culicidae species, describe their abundance and seasonal variation in São José do Rio Preto, SP, and discuss the risk of arbovirus infections. The collection of larval and adult mosquitoes was conducted monthly from 2006 to 2007 in an urban area and four sections of forested land. In the urban area, larvae were collected from sites where oviposition by Culex mosquitoes was most likely to occur. At two of the four sites in the forested land, adult mosquitoes were collected with the use of CDC traps at night, and a Nasci aspirator was used in the daytime at the two other collection sites. In the urban area, 34 Culicidae species were identified out of a total sample of 8,683 specimens; of these specimens, 80.7% were Culex quinquefasciatus, 9.6% were Culex coronator, 3.2% were Aedes albopictus, and 1.1% were Ochlerotatus fluviatilis. The abundance of Cx. quinquefasciatus larvae was negatively related to rainfal. In the woods, 2,268 mosquitoes were collected, representing 10 genera and 46 species. The most abundant mosquito species were Aedeomyia squamipennis, Culex coronator, Culex (Mel.) Melanoconion section, Culex declarator, Ochlerotatus scapularis, Anopheles triannulatus, Culex bidens/interfor and Culex habilitator/pseudojhantinosoma. The abundance of Cx. quinquefasciatus in the urban area and the presence of other Culicidae species in urban areas and forested land point to the possibility of the transmission of West Nile virus and other arbovirus infections in São José do Rio Preto and other cities. Thus, the enacting of measures aimed at the surveillance of these arbovirus infections is essential.

Diversity of mosquito species was evaluated in different habitats before and after the Igarapava reservoir flooding in the Grande River, Southern Cerrado of Brazil. We aimed at verifying changes in these mosquito populations in consequence of the lake formation. Four habitats were selected as sampling stations: peridomiciliary habitat, pasture, \"veredas\" and gallery forest patch. Bimonthly collections were made with the Shannon trap and human bait, including diurnal, crepuscular and nocturnal period of mosquito activity. The Shannon Index results from the potential vectors were compared using Student t-test. Aedes scapularis, Anopheles darlingi and An. albitarsis senso latu seasonal abundance were described with moving average and compared using chi2 test. There were changes in the mosquito frequency in the habitats, except for the \"veredas\" that was 13 km away from the catchment area. The altering in mosquito species seasonal abundance suggests breeding places expansion. Diversity indexes can be used to monitor changes in mosquito vector population in environments where abrupt disturbance can alter disease transmission cycles. Avaliou-se a diversidade de mosquitos em diferentes habitats antes e depois da formação do reservatório de Igarapava no Rio Grande, Cerrado do Sudeste do Brasil, com objetivo de verificar alteração nessas populações de mosquitos. Quatro habitats foram selecionados como sítios de coleta: a) peridomicílio rural, b) pastagem, c) veredas e d) mata ciliar. Realizaram-se coletas bimestrais com isca humana e armadilha de Shannon, incluindo períodos de atividade culicídea diurna, crepuscular vespertina e noturna. Os resultados do Índice de Shannon para os potenciais vetores nos diferentes habitats foram comparados pelo teste t de Student. As abundâncias sazonais de Aedes scapularis, Anopheles darlingi e Anopheles albitarsis s.l., foram descritas com a média móvel e comparadas com o teste chi2. Houve alteração nas freqüências dos mosquitos nos habitats, com exceção das veredas, que estavam afastadas 13 km da área de influência do reservatório. A modificação nas abundâncias sazonais dos potenciais vetores sugere o incremento de criadouros após a formação do reservatório. Índices de diversidade são ferramentas úteis para monitorar populações de mosquitos vetores em ambientes cujas mudanças drásticas podem alterar ciclos de transmissão de doenças transmitidas por vetores.