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Microculex is a subgenus of Culex that is mainly associated with natural breeding sites in wild environments such as cut bamboos, tree holes and bromeliads. However, recent findings of Culex (Microculex) species in bromeliads in urban areas and in artificial breeding sites close to human habitations suggest that they are becoming more tolerant of urbanization. Whilst dispersion studies of this subgenus have shown the ecological valency of some species in relation to human-impacted areas, there is a scarcity of studies on the blood-feeding habits, vector capacity and taxonomy of Microculex. As all the information about this subgenus is fragmented and restricted in most cases to physical libraries, this review seeks to compile all the information on Microculex published over the last century and a half so that this can be more easily consulted by specialists interested in the subject. The ability of some species of this subgenus to adapt to human-impacted environments by colonizing artificial breeding sites and thus become a potential vector of animal and human diseases is also discussed.

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Culex quinquefasciatus is a cosmopolitan species distributed throughout tropical and subtropical areas of the world. The species is of great epidemiological importance as it is responsible for vectoring the causative agent of lymphatic filariasis and several arboviruses, including West Nile virus. Wing geometric morphometrics has been widely used to assess phenotypic variations in mosquito species. Here, we hypothesize that Cx. quinquefasciatus populations in urban parks in the city of São Paulo, Brazil, have been subjected to anthropogenic selective pressures that are responsible for driving their ecology and behavior. Mosquitoes were collected by CDC traps in five municipal parks in the city of São Paulo. Eighteen anatomical landmark coordinates on each female right wing were digitized. Canonical variate analysis, wireframe graphs, cross-validated reclassification tests and the neighbor-joining method were used to assess phenotypical dissimilarity in wing shape between populations. Centroid size was calculated to assess differences in wing size between populations, which can result from different environmental conditions during immature mosquito development. Moderately heterogeneous wing shape and wing size patterns were found in the populations analyzed, indicating that selective pressures in the urban environment are affecting the wing patterns of Cx. quinquefasciatus populations in the city of São Paulo, Brazil.

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The physicochemical parameters of water, such as pH, salinity, conductivity, and total dissolved solids, can influence mosquito larval development, survival, and abundance. Therefore, it is important to elucidate how these factors influence mosquito occurrence. We hypothesized that the occurrence and community composition of immature mosquito species are driven not only by the availability of suitable aquatic habitats, but also by the physicochemical factors of these habitats. The primary objective of this study was therefore to investigate the influence of the physicochemical parameters of water in different types of aquatic habitats on the occurrence of mosquito species in two remnants of Atlantic Forest in the city of São Paulo, Brazil. Collections of immature mosquitoes and assessment of the physicochemical characteristics of the water in the collection sites were carried out for twelve months. The variation in species composition and occurrence with the measured physicochemical parameters and the type of breeding site was assessed using constrained ordination methods. The results indicate that there was a statistically significant difference in species composition as a function of the different types of aquatic habitats, and that pH had an influence on species occurrence even when the variance explained by the type of aquatic habitat was removed from the analysis. There was a statistically significant association between mosquito species occurrence and pH and salinity, and the former had a significant influence on the mosquito species collected regardless of the type of aquatic habitat, showing that the pH of the breeding site water is an important factor in driving mosquito population dynamics and species distribution.

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São Paulo is one of the largest cities in the world and has several characteristics that favor a diversity of urban and wild mosquitoes. Little is known about how variations in mosquito diversity and feeding preferences for different hosts in different vegetation strata can influence the risk of pathogen transmission to humans. We investigated vertical stratification of mosquitoes and its relationship with vertebrate hosts in environments with different degrees of conservation in two conservation units in the city of São Paulo. Adult mosquitoes were collected using CDC traps, aspiration and Shannon traps. After morphological identification, host blood in engorged females was analyzed by PCR with a vertebrate-specific primer set based on mitochondrial cytochrome b DNA of vertebrates commonly found in the two conservation units. Although a higher abundance of the species Anopheles cruzii and Culex nigripalpus was found in the canopy, blood not only from birds but also from humans and rodents was identified in these mosquitoes. In one of the units, Wyeomyia confusa and Limatus durhamii were found occupying mainly niches at ground level while Culex vaxus was frequently found in the canopy. Haemagogus leucocelaenus, the main vector of yellow fever, was found in low abundance at all collection points, particularly in the canopy. Species richness and composition tended to vary little between canopy and ground level in the same environment, but the abundance between canopy and ground level varied more depending on the species analyzed, the most abundant and frequent species exhibiting a predilection for the canopy. Even those mosquito species observed more frequently in the canopy did not show an association with hosts found in this stratum as most of the blood identified in these species was from humans, suggesting opportunist feeding behavior, i.e., feeding on the most readily available host in the environment. The two most common species in the study, An. cruzii and Cx. nigripalpus, may be able to act as bridge vectors for pathogens to circulate between the forest canopy and ground level.

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Aedes albopictus and Aedes scapularis are vectors of several arboviruses, including the dengue, chikungunya, and Rocio virus infection. While Ae. albopictus is a highly invasive species native to Asia and has been dispersed by humans to most parts of the world, Ae. scapularis is native to Brazil and is widely distributed in the southeast of the country. Both species are highly anthropophilic and are often abundant in places with high human population densities. Because of the great epidemiological importance of these two mosquitoes and the paucity of knowledge on how they have adapted to different urban built environments, we investigated the microgeographic population structure of these vector species in the city of São Paulo, Brazil, using wing geometric morphometrics. Females of Ae. albopictus and Ae. scapularis were collected in seven urban parks in the city. The right wings of the specimens were removed and digitized, and eighteen landmarks based on vein intersections in the wing venation patterns were used to assess cross-sectional variation in wing shape and size. The analyses revealed distinct results for Ae. albopictus and Ae. scapularis populations. While the former had less wing shape variation, the latter had more heterogeneity, indicating a higher degree of intraspecific variation. Our results indicate that microgeographic selective pressures exerted by different urban built environments have a distinct effect on wing shape patterns in the populations of these two mosquito species studied here.

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Cantareira State Park (CSP) is located in the Metropolitan Region of São Paulo, one of the most densely populated areas on the planet. Recently, a yellow-fever epidemic practically annihilated the howler monkey population in this park, and human infections were reported in the vicinity. As simian and human plasmodia also circulate in CSP, the present study sought to provide an update on the mosquito fauna in this park, including an analysis of the diversity in areas with different degrees of conservation and a comparison of the yields achieved with different collection techniques. From October 2015 to April 2017, adult mosquitoes were collected with CDC traps, hand-held battery-powered aspirators and Shannon traps, and larvae and pupae were collected with larval dippers and suction samplers in natural and artificial breeding sites. In total, 11,038 specimens distributed in 103 taxa represented by 16 genera were collected. Both the observed species richness and diversity were greater in the environments with the highest degree of preservation. The 'wild' (most preserved) area in CSP had the greatest species richness, followed by the transition area and human-impacted area. The estimated richness indicated that the three environments may have a greater number of species than observed in this study, and Sorensen's index showed that the average degree of similarity varies little between areas. In the inventory of local species, the Shannon trap was the most efficient collection technique for adult mosquitoes, and the suction sampler the most efficient for immatures. The results highlight the increase in the number of different taxa collected as different mosquito capture techniques were included, confirming the importance of using several strategies to sample the local mosquito fauna satisfactorily when exploring a greater number of ecotopes. CSP is a refuge and shelter for native and introduced mosquito species where new biocenoses including pathogens, vertebrate hosts and vectors can form, allowing zoonotic outbreaks in the local human population to occur.

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Water's physical and chemical characteristics are important constraints in aquatic ecosystems, acting on the development, survival, and adaptation of different organisms. Immature forms of mosquitoes develop in widely diverse aquatic environments and are mainly found in permanent or temporary freshwater bodies with little or no movement. The current study aimed to investigate whether variations in larval habitats' pH, salinity, dissolved oxygen, and water temperature influence the composition of Culicidae assemblages and the presence and abundance of Aedes albopictus and Ae. aegypti. From August 2012 to July 2013, captures of immature forms and measurement of water's physical and chemical profiles were performed monthly in natural and artificial breeding sites in four urban parks in the city of São Paulo, Brazil. Changes in species composition related to the parameters' variation were assessed by multivariate analysis. Regression trees were performed to evaluate the effect of breeding sites' physical and chemical variations on the presence and abundance of Ae. albopictus and Ae. aegypti. The observations suggest ranges of conditions for the measured variables in which most species tend to be found more frequently, and pH and salinity are the variables most closely associated with variations in mosquito composition. Ae. aegypti and Ae. albopictus were present in both natural and artificial breeding sites and were observed under significantly varying conditions of pH, salinity, dissolved oxygen, and temperature. For Ae. albopictus, larval habitat type and pH were the best predictors of incidence and abundance. For Ae. aegypti, pH and salinity were the best predictors of abundance, while dissolved oxygen and larval habitat type were better predictors of presence. This information broadens our understanding of the ecology and interaction of the investigated species with abiotic factors in the aquatic environments, providing useful data for studies that seek to elucidate the underlying mechanisms of selection and colonization of breeding sites by these mosquitoes. This study also reinforces previous observations indicating that Ae. albopictus and Ae. aegypti can colonize diverse types of larval habitats with widely varying physical and chemical conditions.

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Microculex is a subgenus of wild mosquitoes belonging to genus Culex, closely related to preserved environments. Its immature forms are generally associated with natural breeding sites, especially bromeliads. Recent years have witnessed the presence of some Microculex species in anthropic environments, including immature forms in artificial breeding sites, which may represent an adaptive tendency. This study aimed to investigate the variation in the abundance and dispersal of Microculex species in environments with different forest cover proportions. Three sites with different proportions of plant cover (60%, 70%, and 90%) were selected in an environmental protection area in the city of São Paulo, with varying degrees of modification and human presence. Collection was performed from March 2015 to April 2017, targeting bromeliads and artificial containers. Variations in the species' richness, composition, and abundance in different environments were analyzed. Variations in mean abundance and larval density between the different forest cover gradients were analyzed with generalized linear mixed-effects models. A total of 1,028 specimens belonging to 14 species were collected. Richness and composition were similar across the environments. Culex (Mcx.) imitator and Cx. (Mcx.) pleuristriatus were the most abundant species. The results showed a relationship between forest cover reduction and an increase in larval abundance and density for Cx. (Mcx.) pleuristriatus. Cx. (Mcx.) imitator showed a decrease in larval abundance related to a reduction in forest cover. Species from Pleuristriatus Series were found both in bromeliads and in artificial breeding sites. The findings emphasize that the Microculex Group of mosquitoes is not limited to preserved environments, but that some species such as Cx. (Mcx.) pleuristriatus appear to be well-adapted to areas impacted by human activity, colonizing artificial breeding sites, which may suggest a possible domiciliation process.

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BACKGROUND: The mosquito Anopheles (Kerteszia) cruzii is the main vector of human and simian malaria in the Atlantic Forest. This species is usually abundant in the forests where it occurs, preferring to live and feed on canopies, behaviour known as acrodendrophily. However, in several studies and locations this species has been observed in high density near the ground in the forest. In this study, it was hypothesized that factors associated with anthropogenic landscape changes may be responsible for the variation in abundance and acrodendrophily observed in An. cruzii. METHODS: The study was conducted in a conservation unit in the city of São Paulo, Brazil. Monthly entomological collections were performed from March 2015 to April 2017, and the resulting data were used with data from another study conducted in the same area between May 2009 and June 2010. Mosquitoes were collected from five sites using CDC and Shannon traps. Landscape composition and configuration metrics were measured, and generalized linear mixed-effect models were used to investigate the relationship between these metrics and variations in the abundance and acrodendrophily of An. cruzii. RESULTS: The model that showed the best fit for the relationship between landscape metrics and An. cruzii abundance suggests that an increase in the proportion of forest cover leads to an increase in the abundance of this mosquito, while the model that best explained variations in An. cruzii acrodendrophily suggests that an increase in total forest-edge length leads to greater activity by this species at ground level. CONCLUSION: While the data indicate that changes in landscape due to human activities lead to a reduction in An. cruzii abundance, such changes may increase the contact rate between this species and humans living on the edges of forest fragments where An. cruzii is found. Future studies should, therefore, seek to elucidate the effect of these landscape changes on the dynamics of Plasmodium transmission in the Atlantic Forest, which according to some studies includes the participation of simian hosts.

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BACKGROUND: Bromeliads can be epiphytic, terrestrial or saxicolous and use strategies to allow water to be retained in their leaf axils, where various arthropods can be found. These include mosquitoes, whose larvae are the most abundant and commonly found organisms in the leaf axils. The objective of this study was to look for immature forms of mosquitoes (the larval and pupal stages) in bromeliads in municipal parks in São Paulo and to discuss the ecological and epidemiological importance of these insects. METHODS: From October 2010 to July 2013, immature mosquitoes were collected from bromeliads in 65 municipal parks in the city of São Paulo, Brazil, using suction samplers. The immature forms were maintained until adult forms emerged, and these were then identified morphologically. RESULTS: Two thousand forty-two immature-stage specimens belonging to the genera Aedes, Culex, Trichoprosopon, Toxorhynchites, Limatus and Wyeomyia were found in bromeliads in 15 of the 65 parks visited. Aedes albopictus was the most abundant species (660 specimens collected), followed by Culex quinquefasciatus (548 specimens) and Cx. (Microculex) imitator (444). The taxa with the most widespread distribution were Ae. aegypti and Toxorhynchites spp, followed by Ae. albopictus and Cx. quinquefasciatus. CONCLUSION: Bromeliads in urban parks are refuges for populations of native species of Culicidae and breeding sites for exotic species that are generally of epidemiological interest. Hence, administrators and surveillance and mosquito-control agencies must constantly monitor these microenvironments as the presence of these species endangers the health of park users and employees as well as people living near the parks.

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The aim of this work was to investigate whether Haemagogus leucocelaenus and other mosquito species associated with sylvatic transmission of yellow fever virus are present in Cantareira State Park (CSP) in the São Paulo Metropolitan Area (SPMA). From October 2015 to March 2016, adult mosquitoes were captured with the Centers for Disease Control and Prevention traps, manual battery-powered aspirators, and Shannon traps; larvae and pupae were collected in natural and artificial breeding sites. A total of 109 adult mosquito specimens and 30 immature forms belonging to 11 taxonomic categories in 4 genera (Aedes, Psorophora, Sabethes, and Haemagogus) were collected, including Hg. leucocelaenus, the main vector of yellow fever. The entomological findings of the present study indicate that the area is of strategic importance for yellow fever surveillance not only because of the significant numbers of humans and nonhuman primates circulating in CSP and its vicinity but also because it represents a potential route for the disease to be introduced to the SPMA.

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