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.

Resumo Junto à modernidade, a atividade humana tem sido um fator chave ligada aos riscos ambientais globais, as modificações antrópicas em âmbito mundial têm sido causa do surgimento de doenças para os animais silvestres e domésticos, bem como para o ser humano. Em especial, o incremento na distribuição espacial e incidência de doenças infecciosas emergentes (DIE) estão diretamente associados ao desmatamento e às mudanças climáticas globais, além disso, o surgimento de novos agentes de DIE, como o SARS-COV-2, tem sido relatado nos últimos 30 anos. Por outro lado, a biodiversidade tem se mostrado um indicador chave para a saúde dos ecossistemas, além de representar um papel importante na promoção da saúde pública humana. Nas regiões neotropicais, e em especial, no Brasil, várias doenças infecciosas têm demonstrado ser diretamente afetadas pela perda de biodiversidade, como a malária, a síndrome pulmonar por hantavírus, a febre amarela, as arboviroses urbanas, a febre maculosa, entre outras. Para entender melhor a capacidade ecossistêmica de regulação de doenças infecciosas, o programa BIOTA FAPESP tem apoiado pesquisadores e projetos de pesquisa para aumentar o conhecimento sobre a biodiversidade e os ecossistemas brasileiros, como a diversidade de bioagentes de aves, a biodiversidade de animais peçonhentos, a diversidade de espécies de mosquitos em fragmentos florestais dentro de áreas urbanas, a propagação do vírus da febre amarela em território florestal fragmentado, perda e isolamento de remanescentes florestais e a ocorrência de febre maculosa e malária, entre outros. Ressalta-se que o BIOTA FAPESP é um programa de sucesso e deve ser ampliado como importante ferramenta de promoção da saúde pública presente e futura.

Abstract Alongside modernity, the human activity has been a key factor in global environmental risks, with worldwide anthropic modification being the cause of the emergence of diseases for wild and livestock animals, and even humans. In special, the increase in the spatial distribution and in the incidence of some emerging infectious diseases (EID) are directly associated to deforestation and global climate changes. Moreover, the arise of new EID agents, such as the SARS-COV-2 have been reported for the last 30 years. On the other hand, biodiversity has been shown to be a key indicator for ecosystem health, and to pose a role to increase the promotion of human public health. In neotropical regions, and in special, in Brazil, several infectious diseases have been demonstrated to be directly affected for the biodiversity loss, such as malaria, hantavirus pulmonary syndrome, yellow fever, urban arboviruses, spotted fever, amongst other. To better understand the ecosystem capacity of regulation of infectious diseases, FAPESP BIOTA program have supported researchers and research projects to increase knowledge about Brazilian biodiversity and the ecosystems, such as diversity of bird bioagents, venomous animals biodiversity, diversity of mosquitos species in forest patches inside urban areas, propagation of the yellow fever virus over fragmented forest territories, loss of ecological corridors and occurrence of spotted fever and malaria, amongst others. It is noteworthy that FAPESP BIOTA is a successful program and must be expanded as an important tool for present and future public health promotion.

Abstract INTRODUCTION: We aimed to study intraspecific variation in Triatoma costalimai, a potential vector of Chagas disease present in Brazil and Bolivia. METHODS: We analyzed phenotypic (connexivum color patterns, wing morphometrics) and genetic variation (16S mtDNA) of three Brazilian T. costalimai populations. We compared 16S sequences with those of putative Bolivian T. costalimai and its sister species, T. jatai. RESULTS: Brazilian populations had different connexivum color patterns and forewing shapes. A 16S mtDNA haplotype network showed a clear separation of Brazilian T. costalimai from both T. jatai and Bolivian T. costalimai. CONCLUSIONS: We report considerable variability in T. costalimai populations.

Abstract INTRODUCTION This study confirms the occurrence of Psammolestes tertius Lent & Jurberg, 1965 (Hemiptera, Reduviidae, Triatominae) in the state of Paraná, Brazil. METHODS: In 2002, a male specimen of P. tertius was collected in the municipality of Porto Rico, Paraná, Brazil. RESULTS: This finding adds to the data on the geographical distribution of P. tertius from 14 to 15 known occurrences in Brazilian states and, therefore, reports the increase in the diversity of triatomines in Paraná. CONCLUSIONS: The presence of P. tertius in the state of Paraná demonstrates that the biodiversity of these insects may have been underestimated.

ABSTRACT Chloroquine (CQ) and its analog hydroxychloroquine (HCQ) were recently included in several clinical trials as potential prophylactic and therapeutic options for SARS-COV-2 infection/covid-19. However, drug effectiveness in preventing, treating, or slowing the progression of the disease is still unknown. Despite some initial promising in vitro results, rigorous pre-clinical animal studies and randomized clinical trials have not been performed yet. On the other hand, while the potential effectiveness of CQ/HCQ is, at best, hypothetical, their side effects are factual and most worrisome, particularly when considering vulnerable groups of patients being treated with these drugs. in this comment, we briefly explain the possible mechanisms of action of CQ/HCQ for treating other diseases, possible actions against covid-19, and their potent side effects, in order to reinforce the necessity of evaluating the benefit-risk balance when widely prescribing these drugs for SARS-COV-2 infection/covid-19. We conclude by strongly recommending against their indiscriminate use.

ABSTRACT Chloroquine (CQ) and its analog hydroxychloroquine (HCQ) were recently included in several clinical trials as potential prophylactic and therapeutic options for SARS-COV-2 infection/covid-19. However, drug effectiveness in preventing, treating, or slowing the progression of the disease is still unknown. Despite some initial promising in vitro results, rigorous pre-clinical animal studies and randomized clinical trials have not been performed yet. On the other hand, while the potential effectiveness of CQ/HCQ is, at best, hypothetical, their side effects are factual and most worrisome, particularly when considering vulnerable groups of patients being treated with these drugs. in this comment, we briefly explain the possible mechanisms of action of CQ/HCQ for treating other diseases, possible actions against covid-19, and their potent side effects, in order to reinforce the necessity of evaluating the benefit-risk balance when widely prescribing these drugs for SARS-COV-2 infection/covid-19. We conclude by strongly recommending against their indiscriminate use.

ABSTRACT This note reports on occurrences of triatomine species in the city of Sao Paulo, Brazil, registered between 1988 and 2017. Records of triatomines captured in Sao Paulo are based on specimens received spontaneously from Health Surveillance Centers, Health Centers and Zoonosis Control Centers in the city as well as from citizens. Species were identified morphologically at the Public Health Entomology Laboratory, Faculty of Public Health, University of Sao Paulo, where the triatomines, which are vectors of Chagas disease, were tested for Trypanosoma cruzi infection. The first reported occurrence of triatomine bugs in urban Sao Paulo was in 1988. The specimen, which was captured in Jardim Sao Luiz district, was from the genus Panstrongylus and was registered as Panstrongylus sp. but was not sexed. Since this first recorded occurrence, the following species have been found in the city: Panstrongylus geniculatus (2 occurrences), P. megistus (15 occurrences), Triatoma infestans (1 occurrence) and T. sordida (3 occurrences). In this paper, the importance of reporting occurrences of triatomine bugs in the city of Sao Paulo, one of the largest metropolis in the world, is discussed with an emphasis on P. megistus. The occurrences discussed here indicate the importance of entomological surveillance for these vectors even in urban centers although the possibility of vector transmission of Chagas disease in these centers is very low.

ABSTRACT This article reports a list with 912 specimens of the subfamily Triatominae deposited in the Entomological Collection of the Faculty of Public Health of the University of Sao Paulo. The collection is composed of 1 holotype, 3 alotypes, 15 paralectotypes, 77 paratypes, distributed in 5 tribes and 12 genera: Tribus Alberprosenini: genus Alberprosenia Martinez & Carcavallo, 1977; Tribus Bolboderini: genus Microtriatoma Prosen & Martinez, 1952; Tribus Cavernicolini: genus Cavernicola Barber, 1937; Tribus Rhodnini: genus Psammolestes Bergroth, 1941; genus Rhodnius Stal, 1859; Tribus Triatomini: genus Dipetalogaster Usinger 1939; genus Eratyrus Stal 1859; genus Hermanlentia Jurberg & Galvão, 1997; genus Linshcosteus Distant, 1904; 1944; genus Panstrongylus Berg 1879; genus Paratriatoma Barber 1938; genus Triatoma Laporte 1833.

Resumo Parques urbanos do município de São Paulo contêm remanescentes de Mata Atlântica. No sul da cidade há 30 parques municipais, sendo os parques Santo Dias e Shangrila alvos deste estudo. Este estudo teve a proposta de levantamento da fauna de culicídeos desses dois parques no sul da cidade de São Paulo e avaliar a presença de potenciais bioindicadores e espécies vetoras de patógenos aos seres humanos. Os mosquitos foram coletados mensalmente entre março de 2011 e fevereiro de 2012, com aspiradores, armadilhas de Shannon e CDCs para mosquitos adultos e conchas entomológicas e bombas manuais de sucção para os imaturos. O esforço amostral foi avaliado por traçar uma curva de acumulação de espécies, e a riqueza total foi estimada pelo método jackknife de primeira ordem. Para comparar a diversidade entre os dois parques, foram calculados os índices de diversidade de Shannon e de Simpson. A similaridade na composição de espécies foi comparada pelo índice de similaridade de Sorensen. Foram coletados um total de 8.850 espécimes de culicídeos em ambos os parques. Coletas no parque Santo Dias renderam 1.577 mosquitos adultos e 658 imaturos, distribuídos em sete gêneros (Aedes, Anopheles, Culex, Limatus, Mansonia, Toxorhynchites e Wyeomyia) e 27 unidades taxonômicas: Culex nigripalpus, e Aedes fluviatilis foram as mais abundantes unidades taxonômicas coletadas como adultos, enquanto em formas imaturas, as espécies mais abundantes coletadas foram Cx. imitator, Wy. davisi, Wy. galvaoi e Ae. albopictus. Coletas no parque Shangrilá renderam 4.952 espécimes como adultos e 1.663 formas imaturas, distribuídas em oito gêneros (Aedes, Anopheles, Culex, Limatus, Mansonia, Toxorhynchites, Uranotaenia e Wyeomyia) e 36 unidades taxonômicas.. As curvas de acúmulo de espécies em ambos os parques ficaram perto da assíntota, e as estimativas de riqueza total foram próximas às riquezas observadas. Apesar da riqueza observada ter sido maior no parque Shangrilá, não houve diferença estatisticamente significante entre os índices de diversidade mensurados. Em relação à composição de espécies os dois locais compartilharam 16 espécies, incluindo as de maior importância epidemiológica como Culex nigripalpus, Cx. quinquefasciatus, Aedes albopictus e Ae. aegypti. Alguns táxons de culicídeos são bioindicadores de condições ambientais nas áreas ou possuem potencial para veicular patógenos. Atenção deve ser dada a parques urbanos, com inclusão destes locais nos programas oficiais de vigilância entomológica e investigações periódicas na circulação de arbovírus.

Abstract Many parks in the city of São Paulo contain remnants of Atlantic Forest. Of the 30 municipal parks in the South of the city, we investigated two in this study (Santo Dias Park and Shangrilá Park) in order to survey their mosquito fauna and investigate the presence of potential bioindicators of environmental conditions and vectors of human pathogens. Mosquitoes were collected monthly between March 2011 and February 2012 using aspirators, Shannon and CDC traps for adult mosquitoes and larval dippers and suction samplers for immature forms. Sampling effort was evaluated by plotting a species accumulation curve, and total richness was estimated using the first-order jackknife. To compare the diversity between the two parks Shannon and Simpson diversity indexes were calculated. Species similarity was compared by the Sorensen similarity index. In all, 8,850 specimens were sampled in both parks. Collections in Santo Dias Park yielded 1,577 adult mosquitoes and 658 immature individuals distributed in seven genera (Aedes, Anopheles, Culex, Limatus, Mansonia, Toxorhynchites and Wyeomyia) and 27 taxonomic units. Among the adult mosquitoes collected, Culex nigripalpus .and Aedes fluviatilis were the most abundant, while the most abundant immature forms were Cx. imitator, Wy. davisi, Wy. galvaoi and Ae. albopictus. Collections in Shangrilá Park yielded 4,952 adult specimens and 1,663 immature forms distributed in eight genera (Aedes, Anopheles, Culex, Limatus, Mansonia, Toxorhynchites, Uranotaenia and Wyeomyia) and 36 taxonomic units. Species accumulation curves in both parks were close to the asymptote, and the total richness estimate was close to the observed richness. Although the observed species richness was higher in the Shangrilá Park, there was no statistically significant difference between the diversity indexes measured. Regarding species composition, the two sites shared 16 species, including those of epidemiological importance such as Culex nigripalpus, Cx. quinquefasciatus, Aedes albopictus and Ae. aegypti. As some of the mosquito taxa found are bioindicators of environmental conditions and have epidemiological potential to carry pathogens, we recommend that urban parks should be included in official mosquito surveillance programs, and regular surveys carried out to detect circulating arboviruses.

Abstract: INTRODUCTION: This study aimed to investigate Flavivirus infection in birds captured in green areas of São Paulo. METHODS: Oropharyngeal swabs, cloacal swabs, and blood samples from 170 birds captured in two green areas in São Paulo, Brazil were subjected to real time-polymerase chain reaction (RT-qPCR) analysis of Flavivirus specific NS5 gene fragment. RESULTS: All samples were negative for the presence of viral ribonucleic acid (RNA). CONCLUSIONS: Despite the negative results, Flavivirus surveillance must be performed regularly due to favorable ecological conditions for virus circulation and transmission among birds in these areas and their close proximity to humans.

Abstract: INTRODUCTION: Mass production of mosquitoes under laboratory conditions allows implementing methods to control vector mosquitoes. Colony development depends on mosquito size and weight. Body size can be estimated from its correlation with wing size, whereas weight is more difficult to determine. Our goal was to test whether wing size can predict the weight. METHODS: We compared dry weight and wing centroid size of Culex quinquefasciatus reared at different temperatures and four diets. RESULTS: Weight and wing size were strongly correlated. The diets did not influence wing size. CONCLUSIONS: Wing centroid size is a good predictor of Cx. quinquefasciatus body weight.

Em São Paulo os parques municipais preservam resquícios da vegetação de Mata Atlântica, comportam lagos e nascentes, servem de abrigos para mamíferos, aves e artrópodes vetores. São frequentados pela população humana como ambientes de lazer. A zona Leste da cidade possui o maior índice populacional de habitantes e 24 parques municipais. O objetivo deste trabalho foi conhecer a fauna de Culicidae de dois parques da zona Leste da cidade e avaliar a presença de espécies potencialmente bioindicadoras e vetoras de patógenos ao homem. Foram feitas coletas mensais de culicídeos entre março de 2011 a fevereiro de 2012 no Parque do Carmo e no Parque Chico Mendes, com aspiradores elétricos, armadilhas de Shannon e CDC para mosquitos adultos e concha entomológica e bombas manuais de sucção para os imaturos. Para verificar a suficiência amostral foram construídas curvas de acúmulo de espécies por amostragem e a riqueza total foi estimada pelo método Jackknife 1, ambos gerados pelo programa EstimateS. No parque do Carmo foram coletados 1.092 culicídeos, distribuídos em nove gêneros (Aedes, Anopheles, Coquillettidia, Culex, Limatus, Mansonia, Trichoprosopon, Toxorhynchites e Uranotaenia), distribuídos em 19 unidades taxonômicas. Coquillettidia venezuelensis(Theobald 1912), Aedes scapularis (Rondani 1848)e Culex (Culex) spp. Linnaeus 1758 foram os adultos mais abundantes e Culex (Melanoconion) spp. Theobald 1903 e Anopheles strodei Root 1926 os imaturos. No Parque Chico Mendes obteve-se 4.487 mosquitos, distribuídos em seis gêneros e 18 unidades taxonômicas. Culex (Cux.) spp. e Ae. scapularis foram os mais abundantes dos adultos e Ae. albopictus (Skuse 1984) dos imaturos. As curvas de acúmulo de espécies por amostragem em ambos os parques chegaram próximas è assíntota e a estimativa de riqueza total mostrou-se próxima è riqueza observada. Alguns táxons de culicídeos são bioindicadores das condições ambientais dos locais onde vivem, portanto é necessário acompanhar a fauna nativa dos parques municipais porque várias espécies encontradas têm mostrado competência e capacidade para veicular patógenos e arbovírus.

Municipal parks in the city of São Paulo, which are used for leisure purposes, contain remnants of the flora of the Atlantic Forest, as well as lakes and springs, and are home to mammals, birds and arthropods, some of which can be vectors of disease. The eastern side of the city has the largest population in São Paulo and twenty-four municipal parks. The aim of this study was to investigate Culicidae fauna in two parks on this side of the city and to determine which of the Culicidae species identified have the potential to act as bioindicators and vectors of human pathogens. Culicidae were collected monthly between March 2011 and February 2012 in Carmo Park and Chico Mendes Park with battery-powered aspirators, Shannon traps and CDC traps for adult mosquitoes, and larval dippers and suction samplers for immature mosquitoes. To confirm sample sufficiency, the EstimateS program was used to plot sample-based species accumulation curves and estimate total richness by the Jackknife 1 method. In all, 1,092 culicids from nine genera (Aedes, Anopheles, Coquillettidia, Culex, Limatus, Mansonia, Trichoprosopon, Toxorhynchites and Uranotaenia) and nineteen taxonomic units were collected in Carmo Park. Coquillettidia venezuelensis (Theobald 1912), Aedes scapularis (Rondani 1848) and Culex (Culex) spp. Linnaeus 1758 were the most abundant adults, and Culex (Melanoconion) spp. Theobald 1903 and Anopheles strodei Root 1926 the most abundant immature mosquitoes. In Chico Mendes Park 4,487 mosquitoes in six genera and eighteen taxonomic units were collected. Culex (Cux.) spp. and Ae. scapularis were the most abundant adults, and Ae. albopictus (Skuse 1984) the most abundant immature mosquitoes. The species accumulation curves in both parks were close to the asymptote, and the total richness estimate was close to the observed richness. Some culicid taxons are bioindicators of environmental conditions in the areas they inhabit. It is important to monitor native fauna in municipal parks in São Paulo as various species in this study were found to have vector competence and capacity to transmit pathogens, such as arboviruses.

Culex quinquefasciatus mosquitoes have been successfully genetically modified only once, despite the efforts of several laboratories to transform and establish a stable strain. We have developed a transient gene expression method, in Culex, that delivers plasmid DNA directly to the mosquito haemolymph and additional tissues. We were able to express DsRed2 fluorescent protein in adult Cx. quinquefasciatus mosquitoes by injecting plasmids directly into their thorax. The expression of DsRed2 in adult Cx. quinquefasciatus mosquitoes is an important stepping stone to genetic transformation and the potential use of new control strategies and genetic interactions.

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.

Culex quinquefasciatus é vetor de patógenos humanos, incluindo nematódeos filarídeos e vários vírus. Embora a sua relevância epidemiológica varie entre as diferentes regiões geográficas, o conhecimento da estrutura genética da população é ainda incipiente. Em vista disso, foram avaliados os níveis de diversidade genética de Cx. quinquefasciatus e de híbridos Cx. quinquefasciatus x Cx. pipiens de nove cidades do Brasil e em La Plata, na Argentina. Para os testes foram utilizados fragmentos dos genes mitocondriais cox1 e nd4, juntamente com coxA e wsp do endossimbionte Wolbachia, herdado maternalmente. O fragmento nd4 não apresentou variação entre as amostras, e o cox1 exibiu quatro haplótipos que separaram dois tipos de Cx. quinquefasciatus, com um deles agrupado no sul do Brasil. Os dados de sequência mostraram baixa diversidade, sendo esta discutida. Ambas as amostras de mosquitos brasileiros e argentinos estão infectados com uma única cepa de Wolbachia. A diversidade apresentada por nd4 e cox1 não é congruente com a estrutura da população revelada por marcadores nucleares e morfologia alar de estudos anteriores com estas mesmas populações. Pesquisas com Cx. quinquefasciatus devem, se possível, avaliar a diversidade por DNA mitocondrial na luz de outros marcadores.

Culex quinquefasciatus is a vector of human pathogens, including filarial nematodes and several viruses. Although its epidemiological relevance is known to vary across geographical regions, an understanding of its population genetic structure is still incipient. In light of this, we evaluated the genetic diversity of Cx. quinquefasciatus and Cx. pipiens x Cx. quinquefasciatus hybrids collected from nine localities in Brazil and one site in Argentina. We used mitochondrial genes cox1 and nd4, along with the coxA and wsp genes of the maternally-inherited Wolbachia endosymbiont. The nd4 fragment was invariant between samples, whilst cox1 exhibited four haplotypes that separated two types of Cx. quinquefasciatus, one clustered in southern Brazil. Low sequence diversity was generally observed, being discussed. Both Brazilian and Argentinian mosquitoes were infected with a single Wolbachia strain. As reported in previous studies with these populations, cox1 and nd4 diversity is not congruent with the population structure revealed by nuclear markers or alar morphology. Future Cx. quinquefasciatus research should, if possible, evaluate mtDNA diversity in light of other markers.