ABSTRACT Background: Despite their worldwide occurrence, the distribution and role of insect-specific flaviviruses (ISFs) remain unclear. Methods: We evaluated the presence of ISFs in mosquitoes collected in São Paulo, Brazil, using reverse transcription and semi-nested polymerase chain reaction (PCR). Some of the positive samples were subjected to nanopore sequencing. Results: Twelve mosquito pools (2.8%) tested positive for flavivirus infection. Nanopore sequencing was successfully performed on six samples. Phylogenetic analysis grouped these sequences into genotype 2 of Culex flavivirus (CxFV). Conclusions: The identification of CxFV genotype 2 at new locations in São Paulo highlights the importance of understanding the role of ISFs in mosquito vector competence.

ABSTRACT OBJECTIVE: To evaluate locomotor activity in four field populations of Ae. aegypti with different insecticide resistance profiles from the state of São Paulo for two years. METHODS: This study comprised the susceptible Rockefeller strain and four populations from São Paulo, Brazil: two considered populations with “reduced susceptibility” to pyrethroids (Campinas and Marília), and two “resistant populations” (Santos and Ribeirão Preto). First, 2016 and 2017 eggs from these five populations were hatched in laboratory. Virgin females underwent experiments under laboratory conditions at 25°C, with 12:12h light/dark (LD) photoperiod; 24-hour individual activity was recorded using a locomotor activity monitor (LAM). RESULTS: In females from 2016 field populations, both resistant populations showed significant more locomotor activity than the two reduced susceptibility populations and the Rockefeller strain (p < 0.05). As for females from 2017 field populations, reduced susceptibility populations showed a significant increased locomotor activity than the Rockefeller strain, but no significant difference when compared to Santos resistant population (p > 0.05). CONCLUSIONS: Our results indicate that insecticide-resistant Ae. aegypti populations show increased locomotor activity, which may affect the transmission dynamics of their arboviruses.

ABSTRACT OBJECTIVE: To evaluate locomotor activity in four field populations of Ae. aegypti with different insecticide resistance profiles from the state of São Paulo for two years. METHODS: This study comprised the susceptible Rockefeller strain and four populations from São Paulo, Brazil: two considered populations with “reduced susceptibility” to pyrethroids (Campinas and Marília), and two “resistant populations” (Santos and Ribeirão Preto). First, 2016 and 2017 eggs from these five populations were hatched in laboratory. Virgin females underwent experiments under laboratory conditions at 25°C, with 12:12h light/dark (LD) photoperiod; 24-hour individual activity was recorded using a locomotor activity monitor (LAM). RESULTS: In females from 2016 field populations, both resistant populations showed significant more locomotor activity than the two reduced susceptibility populations and the Rockefeller strain (p < 0.05). As for females from 2017 field populations, reduced susceptibility populations showed a significant increased locomotor activity than the Rockefeller strain, but no significant difference when compared to Santos resistant population (p > 0.05). CONCLUSIONS: Our results indicate that insecticide-resistant Ae. aegypti populations show increased locomotor activity, which may affect the transmission dynamics of their arboviruses.

Abstract INTRODUCTION: This study aimed to assess the occurrence of gonotrophic discordance in females of Culex quinquefasciatus in São Paulo, Brazil. METHODS: Resting females were collected monthly for 8 months. Females of Cx. quinquefasciatus were identified, and their midgut and ovaries were dissected. RESULTS: Two hundred females were dissected, out of which, 27.5% were nulliparous and 57% were parous. Most females had no blood in the midgut, but gonotrophic discordance was found in 21% females. CONCLUSIONS: Females of Cx. quinquefasciatus showed a high parity rate and gonotrophic discordance, which could favor the vector capacity of this species.

RESUMO Descreve-se o encontro de criadouro subterrâneo de Aedes aegypti no bairro de Pinheiros, São Paulo, SP, ocorrido durante um programa de levantamento entomológico, realizado em 2016. Mesmo com intensa vigilância e controle vetorial, grande quantidade de mosquitos estava presente nessa área. Investigação minuciosa permitiu a detecção de Ae. aegypti em reservatório subterrâneo para armazenamento de água pluvial. Após a implantação de telas de proteção nos acessos, não foi mais detectada a presença do vetor. O uso frequente desse tipo de estrutura e o seu risco para a produção de mosquitos é discutido.

ABSTRACT We describe the discovery of Aedes aegypti underground breeding site in the Pinheiros neighborhood of São Paulo, SP, during an entomological survey program performed in 2016. Even with intense surveillance and vector control, large numbers of mosquitoes were present in this area. A detailed investigation allowed for the detection of Ae. aegypti in an underground reservoir used for rainwater storage. After the implementation of protection screens in the accesses, the presence of the vector was no longer detected. In this study, we discuss the frequent use of this type of reservoir structure and its risk for mosquito production.

RESUMO Descreve-se o encontro de criadouro subterrâneo de Aedes aegypti no bairro de Pinheiros, São Paulo, SP, ocorrido durante um programa de levantamento entomológico, realizado em 2016. Mesmo com intensa vigilância e controle vetorial, grande quantidade de mosquitos estava presente nessa área. Investigação minuciosa permitiu a detecção de Ae. aegypti em reservatório subterrâneo para armazenamento de água pluvial. Após a implantação de telas de proteção nos acessos, não foi mais detectada a presença do vetor. O uso frequente desse tipo de estrutura e o seu risco para a produção de mosquitos é discutido.

ABSTRACT We describe the discovery of Aedes aegypti underground breeding site in the Pinheiros neighborhood of São Paulo, SP, during an entomological survey program performed in 2016. Even with intense surveillance and vector control, large numbers of mosquitoes were present in this area. A detailed investigation allowed for the detection of Ae. aegypti in an underground reservoir used for rainwater storage. After the implementation of protection screens in the accesses, the presence of the vector was no longer detected. In this study, we discuss the frequent use of this type of reservoir structure and its risk for mosquito production.

RESUMO Descreve-se o encontro de nove larvas de Aedes aegypti em criadouro natural no bairro de Pinheiros, município de São Paulo, SP. O registro ocorreu no mês de dezembro de 2014, durante programa de vigilância entomológica dos vetores do vírus dengue, com busca ativa de potenciais criadouros, artificiais ou naturais. O encontro de larvas de Ae. aegypti em oco de árvore revela a capacidade dessa espécie de utilizar tanto criadouros artificiais como naturais para o seu desenvolvimento, o que aponta para a importância da manutenção de vigilância contínua sobre esse mosquito em todos os tipos de recipientes que podem acumular água.

ABSTRACT This is the description of how nine Aedes aegypti larvae were found in a natural breeding site in the Pinheiros neighborhood, city of Sao Paulo, SP, Southeastern Brazil. The record was conducted in December 2014, during an entomological surveillance program of dengue virus vectors, with an active search of potential breeding sites, either artificial or natural. Finding Ae. aegypti larvae in a tree hole shows this species’ ability to use both artificial and natural environments as breeding sites and habitats, which points towards the importance of maintaining continuous surveillance on this mosquito in all kinds of water-holding containers.

RESUMO Descreve-se o encontro de nove larvas de Aedes aegypti em criadouro natural no bairro de Pinheiros, município de São Paulo, SP. O registro ocorreu no mês de dezembro de 2014, durante programa de vigilância entomológica dos vetores do vírus dengue, com busca ativa de potenciais criadouros, artificiais ou naturais. O encontro de larvas de Ae. aegypti em oco de árvore revela a capacidade dessa espécie de utilizar tanto criadouros artificiais como naturais para o seu desenvolvimento, o que aponta para a importância da manutenção de vigilância contínua sobre esse mosquito em todos os tipos de recipientes que podem acumular água.

ABSTRACT This is the description of how nine Aedes aegypti larvae were found in a natural breeding site in the Pinheiros neighborhood, city of Sao Paulo, SP, Southeastern Brazil. The record was conducted in December 2014, during an entomological surveillance program of dengue virus vectors, with an active search of potential breeding sites, either artificial or natural. Finding Ae. aegypti larvae in a tree hole shows this species’ ability to use both artificial and natural environments as breeding sites and habitats, which points towards the importance of maintaining continuous surveillance on this mosquito in all kinds of water-holding containers.

INTRODUCTION: Aedes albopictus was first detected in Brazil in 1986. This mosquito species presents a major threat to public health because Brazilian populations have shown substantial vector competence for arboviruses such as dengue and chikungunya. METHODS: We updated the records of Ae. albopictus in several States of Brazil, focusing on areas in which its presence had been reported after 2002. RESULTS: Twenty-eight years after its arrival in Brazil, Ae. albopictus has been detected in 24 of 27 States. CONCLUSIONS: The rapid spread of this species and its high vector competence demonstrate the danger of Ae. albopictus in Brazil.

Diante da escassez de informações sobre mosquitos (Diptera: Culicidae) na cidade de São Paulo, foi proposto um projeto para estudar esse grupo de insetos nas áreas verdes representadas pelos parques municipais da cidade. Foram investigados 35 desses parques distribuídos nas regiões sul, norte e centro-oeste da cidade, entre outubro de 2010 e fevereiro de 2011 em período diurno. Os imaturos foram coletados dos criadouros por meio de conchas entomológicas e bomba de sucção e os adultos foram capturados em seus abrigos por aspirador elétrico (bateria de 12V). A identificação e catalogação de espécimes foram feitas no Laboratório de Entomologia da Faculdade de Saúde Pública, Universidade de São Paulo. Nesta primeira fase do projeto, coletou-se um total de 5.129 espécimes distribuídos em 11 gêneros e 41 categorias taxonômicas. Culex (Cux.) quinquefasciatus foi a espécie mais abundante. O gênero Aedes foi representado principalmente por Ae. (Och.) fluviatilis e Ae. (Ste.) albopictus. Ae. (Ste.) aegypti e Ae. (Och.)scapularis também foram frequentes em alguns parques. Os demais gêneros apresentaram-se pouco abundantes. Dos parques, 25,7% apresentaram mais de dez grupos, com destaque para o Anhanguera com 26; em contrapartida, 57,1% apresentaram cinco ou menos grupos. Apesar da pressão antrópica sobre esses ambientes, diversas espécies de culicídeos se utilizam destes habitats para a manutenção e refúgio de suas populações. É recomendado que estes ambientes estejam sob constante vigilância epidemiológica, visto que algumas das espécies coletadas possuem importância em saúde pública como vetoras de patógenos à população humana.

Given the scarcity of information on mosquitoes (Diptera: Culicidae) in the city of São Paulo, led the authors to propose a project to investigate this group of insects in some of the green areas, represented by municipal parks. The captures were undertaken in 35 municipal parks in the south, north and central-west of the city, between October 2010 and February 2011, during daylight. Immature forms were collected from breeding places with entomological ladles and suction pumps and adults from resting places with electric aspirators (12V battery). The identification of the specimens was undertaken in the Culicid laboratory of the Faculdade de Saúde Pública, Universidade de São Paulo. In this first phase of the project, a total of 5,129 specimens distributed in 11 genera and 41 taxonomic categories were captured. Culex (Cux) quinquefasciatus was the most abundant species. The genus Aedes was represented mainly by Ae. (Och) fluviatilis and Ae. (Ste) albopictus. Ae. (Ste) aegypti and Ae. (Och) scapularis were frequent in some parks. Other genera were less abundant. Of the parks investigated, 25.7% presented more than ten groups of species, notably the Anhanguera with 26; on the other hand, 57.1% had five or fewer groups. Despite the anthropic pressure on these environments, several culicid species make use of these habitats as refuges. It is recommended that these environments be kept under constant epidemiological surveillance as some of the species collected constitute public health threats as pathogen vectors to the human population.

Este estudo contribui para o conhecimento de espécies de Anopheles, incluindo vetores de Plasmodium do oeste da Amazônia brasileira, em Porto Velho, no estado de Rondônia. Esta região vem passando por mudanças ambientais, como consequência de agricultura extensiva e projetos hidroelétricos que causam desmatamento, favorecendo o desenvolvimento de algumas espécies de mosquitos. Assim, a proposta deste estudo é registrar a presença de espécies de anofelinos na área, sendo conduzidas coletas de mosquitos em três locais, ao longo de uma linha de transmissão de energia elétrica. Cada uma das localidades foi amostrada três vezes, no período de 2010 a 2011. Os principais mosquitos adultos capturados em armadilhas de Shannon foram Anopheles darlingi, An. triannulatus, An. nuneztovari l.s., An.gilesi e An. costai. Assim como as formas larvárias Anopheles braziliensis, An. triannulatus, An. darlingi, An. deaneorum, An. marajoara, An. peryassui, An. nuneztovari l.s. e An. oswaldoi-konderi, coletadas em criadouros. Anopheles darlingi foi a espécie mais coletada na região. Em adição, discutiu-se sistemática de Culicidae, distribuição de fauna e aspectos da malária em ambientes modificados do oeste da Amazônia brasileira.

This study contributes to knowledge of Anopheles species, including vectors of Plasmodium from the western Brazilian Amazon in Porto Velho, Rondônia State. The sampling area has undergone substantial environmental changes as a consequence of agricultural and hydroelectric projects, which have caused intensive deforestation and favored habitats for some mosquito species. The purpose of this study was to diagnose the occurrence of anopheline species from collections in three locations along an electric-power transmission line. Each locality was sampled three times from 2010 to 2011. The principal adult mosquitoes captured in Shannon trap were Anopheles darlingi, An. triannulatus, An. nuneztovari l.s., An.gilesi and An. costai. In addition, larvae were collected in ground breeding sites for Anopheles braziliensis, An. triannulatus, An. darlingi, An. deaneorum, An. marajoara, An. peryassui, An. nuneztovari l.s. and An. oswaldoi-konderi. Anopheles darlingi was the most common mosquito in the region. We discuss Culicidae systematics, fauna distribution, and aspects of malaria in altered habitats of the western Amazon.

INTRODUÇÃO: Alterações no ambiente vêm contribuindo com mudanças climáticas, como o aumento do volume de chuvas, que acarreta as inundações. Medidas estão sendo tomadas no enfrentamento das inundações, como a implantação dos reservatórios de contenção de cheias (piscinões). Neste trabalho, foi avaliada a fauna de culicídeos, de importância epidemiológica, nos piscinões Caguaçu e Inhumas. MÉTODOS: Foram realizadas coletas mensais nos piscinões Caguaçu e Inhumas, situados na região leste de São Paulo, de março de 2006 a fevereiro de 2007, empregando-se os métodos de concha entomológica e aspirador. Para análise dos dados, foram realizadas análises estatísticas descritivas e a regressão linear simples. RESULTADOS: Foram coletados 8.917 culicídeos, destacando-se Culex (Culex) quinquefasciatus, que representou 98,9% dos espécimes no Inhumas e 95,2% no Caguaçu. No Caguaçu, a maior frequência de imaturos foi observada no vertedouro (61%) e no Inhumas na canaleta (42,6%). A precipitação prediz 87% da abundância numérica de larvas de terceiro e de quarto estágio no Caguaçu e 60% do número de pupas coletadas. No Inhumas, a precipitação explicou 36% da abundância numérica de larvas e 18% do número de pupas. CONCLUSÕES: Culex quinquefasciatus, vetor de agentes da filariose, arboviroses e fator de incômodo à população, foi a espécie mais frequente nos dois ambientes. Medidas de controle da espécie nos piscinões estudados se fazem necessárias tendo em vista seu potencial epidemiológico.

INTRODUCTION: Alterations in the environment contribute to changes in weather patterns, which cause an increase in rainfall, which causes flooding. Flooding has been addressed by building reservoirs, called piscinões, to contain the excess rain. In this report, we evaluated the Culicidae fauna of epidemiological importance in the reservoirs of Caguaçu and Inhumas. METHODS: Monthly collections were performed in both reservoirs, which are situated in the Eastern section of the City of São Paulo. Monthly collections of Culicidae were undertaken using the entomological scoop method and a battery aspirator during the period between March 2006 and February 2007. Descriptive statistical and simple linear regression analyses of the data were carried out. RESULTS: A total of 8,917 Culicidae mosquitoes were collected. The majority of the insects captured were of Culex (Culex) quinquefasciatus, representing 98.9% of the specimens identified in the Inhumas and 95.2% of those identified in Caguaçu. In the Caguaçu reservoir, the greatest frequency of immature mosquitoes was observed in the spillway (61%). In the Inhumas reservoir, the greatest occurrence of immature Culicidae was detected in the channel (42.6%). The linear regression analysis showed that 87% of the larvae in the third and fourth stages and 60% of the pupae were collected during periods of rainfall. In the Inhumas reservoir, 36% of the larvae and 18% of the pupae were collected during periods of rainfall. CONCLUSIONS: Culex quinquefasciatus, a vector for agents of filariasis and arbovirus diseases and a nuisance to the human population, was frequent in both environments. Methods of control of the species in the studied dams are necessary to reduce their epidemiological potential.

Em pesquisa de campo realizada no interior do Estado do Espírito Santo, Brasil, em janeiro de 2006, como parte de projeto sobre a transmissão de Plasmodium, foram coletadas larvas de anofelinos em bromélias. Os imaturos foram mantidos no laboratório até a obtenção dos adultos machos e fêmeas associados com as exúvias das larvas e das pupas, para serem identificados. Conseqüentemente, verificou-se que dois espécimes pertenciam a Anopheles (Kerteszia) homunculus Komp, 1937. Este é o primeiro registro dessa espécie de Kerteszia no Espírito Santo. O encontro evidencia a importância de estudos adicionais de modo a estabelecer a distribuição geográfica do An. homunculus, bem como o status taxonômico e a importância epidemiológica da espécie na dinâmica da transmissão da malária em áreas de Mata Atlântica.

In a field collecting trip carried out in the State of Espírito Santo, Brazil, in January 2006, as part of a study on Plasmodium transmission, Anophelinae larvae were collected in bromeliads. The immatures were raised in the laboratory until adult stage, and thus adults male and female and their associated larval and pupal exuviae were mounted and identified. Consequently, it was verified that two specimens belong to Anopheles (Kerteszia) homunculus Komp, 1937. This is the first record of that Kerteszia species in Espírito Santo. The present finding shows the importance of additional studies aiming to establish the geographical distribution of An. homunculus in addition to the taxonomic status and the epidemiological importance of the species in the dynamics of malaria transmission in areas of Atlantic Forest.

Empreenderam-se coletas de imaturos e adultos de Culicidae no Vale do Rio Doce, Estado do Espírito Santo, Brasil, com o objetivo de ampliar o conhecimento sobre os anofelíneos dessa região. Do material obtido, além de outros anofelíneos e demais mosquitos, destaca-se a presença de Anopheles darlingi, An. oswaldoi e An. costai. Quanto ao An. darlingi, responsável por surtos de malária nas regiões baixas daquele Estado, já se esperava a confirmação de sua presença. Assinala-se como relevante a captura de espécimes de An. costai, ampliando-se a distribuição geográfica da espécie, bem como, trazendo contribuição para o conhecimento da Serie Arribalzagia do subgênero Anopheles.

Field collections of immatures and adults of Culicidae were carried out in the Rio Doce Valley, Espírito Santo State, aiming to increase knowledge on the anopheline species of that region. Considering all Culicidae species collected, among other anopheline it is noteworthy the presence of Anopheles darlingi, An. oswaldoi and An. costai. Regarding to An. darlingi, responsible for malaria outbreak in the low lands in that state, it was expected to confirm its presence in the area. The register of specimens of An. costai in the field collections is relevant, increasing the geographical distribution of the species and representing better knowledge of the Series Arribalzagia of the subgenus Anopheles.

O represamento do Rio Pananá para construção da hidrelétrica de Porto Primavera, entre os Estados do Mato Grosso do Sul e São Paulo, alterou as relações ecológicas na região. O objetivo do estudo foi descrever a fauna de culicídeos potencialmente vetores nesse reservatório, a 2km da margem direita, em Bataguassu, Mato Grosso do Sul, na fase anterior à inundação. Os culicídeos foram capturados em ambientes distintos mensalmente, de julho de 1997 a novembro de 1999. Foram calculados índices de riqueza e abundância. Obtiveram-se 16.553 exemplares adultos e 1.795 imaturos, com riqueza de 86 e 44 espécies, respectivamente. A fauna culicídea da área de estudo apresentava relativa riqueza, com espécies de valor epidemiológico, como o Anopheles darlingi, além de outras com potencial na veiculação de arbovírus.

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.