Gabriela Cristina de Carvalho

Mosquito Fauna in Municipal Parks of São Paulo City, Brazil: A Preliminary Survey

Abstract 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.

Diversity and abundance of mosquitoes (Diptera:Culicidae) in an urban park: larval habitats and temporal variation.

Urban parks are areas designated for human recreation but also serve as shelter and refuge for populations of several species of native fauna, both migratory and introduced. In Brazil, the effect of annual climate variations on Aedes aegypti and dengue epidemics in large cities like São Paulo is well known, but little is known about how such variations can affect the diversity of mosquito vectors in urban parks and the risk of disease transmission by these vectors. This study investigates the influence of larval habitats and seasonal factors on the diversity and abundance of Culicidae fauna in Anhanguera Park, one of the largest remaining green areas in the city of São Paulo. Species composition and richness and larval habitats were identified. Seasonality (cold-dry and hot-rainy periods) and year were considered as explanatory variables and the models selection approach was developed to investigate the relationship of these variables with mosquito diversity and abundance. A total of 11,036 specimens from 57 taxa distributed in 13 genera were collected. Culex nigripalpus, Cx. quinquefasciatus and Aedes albopictus were the most abundant species. Bamboo internodes and artificial breeding sites showed higher abundance, while ponds and puddles showed greater richness. Significant relationships were observed between abundance and seasonality, with a notable increase in the mosquitos abundance in the warm-rainy periods. The Shannon and Berger-Parker indices were related with interaction between seasonality and year, however separately these predictors showed no relationship with ones. The increased abundance of mosquitoes in warm-rainy months and the fact that some of the species are epidemiologically important increase not only the risk of pathogen transmission to people who frequent urban parks but also the nuisance represented by insect bites. The findings of this study highlight the importance of knowledge of culicid ecology in green areas in urban environments.

Blood meal sources of mosquitoes captured in municipal parks in São Paulo, Brazil

ABSTRACT: The aim of this study was to investigate blood meal sources of mosquitoes captured in municipal parks in the city of São Paulo, Brazil, and to identify possible associations between mosquito species and their food preferences. Fourteen species of blood hosts of 510 engorged adult female mosquitoes were identified using PCR assays with a vertebrate-specific primer set based on cytochrome b mitochondrial DNA of the following vertebrates: birds, dogs, cats, rodents, humans, and other primates. Mosquitoes were captured using a manual aspirator, CDC traps in the canopy, CDC traps at ground level, and Shannon traps. With the exception of cats, all other vertebrates were used as hosts by mosquitoes in the parks. Statistical analysis failed to show any trend toward association between most culicid species captured and the sources of blood meals. Instead, they revealed random patterns, indicating that the mosquitoes fed on the most abundant or convenient blood meal sources. Although feeding preferences were observed in two species (birds in the case of Cx. nigripalpus and dogs in the case of Cx. quinquefasciatus), our results highlight the opportunistic feeding habits of the female mosquitoes in this study.

Morphometric Wing Characters as a Tool for Mosquito Identification

Mosquitoes are responsible for the transmission of important infectious diseases, causing millions of deaths every year and endangering approximately 3 billion people around the world. As such, precise identification of mosquito species is crucial for an understanding of epidemiological patterns of disease transmission. Currently, the most common method of mosquito identification relies on morphological taxonomic keys, which do not always distinguish cryptic species. However, wing geometric morphometrics is a promising tool for the identification of vector mosquitoes, sibling and cryptic species included. This study therefore sought to accurately identify mosquito species from the three most epidemiologically important mosquito genera using wing morphometrics. Twelve mosquito species from three epidemiologically important genera (Aedes, Anopheles and Culex) were collected and identified by taxonomic keys. Next, the right wing of each adult female mosquito was removed and photographed, and the coordinates of eighteen digitized landmarks at the intersections of wing veins were collected. The allometric influence was assessed, and canonical variate analysis and thin-plate splines were used for species identification. Cross-validated reclassification tests were performed for each individual, and a Neighbor Joining tree was constructed to illustrate species segregation patterns. The analyses were carried out and the graphs plotted with TpsUtil 1.29, TpsRelw 1.39, MorphoJ 1.02 and Past 2.17c. Canonical variate analysis for Aedes, Anopheles and Culex genera showed three clear clusters in morphospace, correctly distinguishing the three mosquito genera, and pairwise cross-validated reclassification resulted in at least 99% accuracy; subgenera were also identified correctly with a mean accuracy of 96%, and in 88 of the 132 possible comparisons, species were identified with 100% accuracy after the data was subjected to reclassification. Our results showed that Aedes, Culex and Anopheles were correctly distinguished by wing shape. For the lower hierarchical levels (subgenera and species), wing geometric morphometrics was also efficient, resulting in high reclassification scores.

Mosquito (Diptera: Culicidae) fauna in parks in greater São Paulo, Brazil

Municipal parks in the city of Sao 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 Sao 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 Sao Paulo as various species in this study were found to have vector competence and capacity to transmit pathogens, such as arboviruses.

Biodiversidade de mosquitos (Diptera: Culicidae) nos parques da cidade de São Paulo I

Given the scarcity of information on mosquitoes (Diptera: Culicidae) in the city of Sao 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 Saude Publica, Universidade de Sao 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.

Wing variation in Culex nigripalpus (Diptera: Culicidae) in urban parks

BackgroundCulex nigripalpus has a wide geographical distribution and is found in North and South America. Females are considered primary vectors for several arboviruses, including Saint Louis encephalitis virus, Venezuelan equine encephalitis virus and Eastern equine encephalitis virus, as well as a potential vector of West Nile virus. In view of the epidemiological importance of this mosquito and its high abundance, this study sought to investigate wing variation in Cx. nigripalpus populations from urban parks in the city of São Paulo, Brazil.MethodsFemale mosquitoes were collected in seven urban parks in the city of São Paulo between 2011 and 2013. Eighteen landmark coordinates from the right wing of each female mosquito were digitized, and the dissimilarities between populations were assessed by canonical variate analysis and cross-validated reclassification and by constructing a Neighbor-Joining (NJ) tree based on Mahalanobis distances. The centroid size was calculated to determine mean wing size in each population.ResultsCanonical variate analysis based on fixed landmarks of the wing revealed a pattern of segregation between urban and sylvatic Cx. nigripalpus, a similar result to that revealed by the NJ tree topology, in which the population from Shangrilá Park segregated into a distinct branch separate from the other more urban populations.ConclusionEnvironmental heterogeneity may be affecting the wing shape variation of Cx. nigripalpus populations.

Microsatellite Markers Transferability in Culex nigripalpus (Diptera: Culicidae)

Abstract Microsatellite loci specifically designed and optimized for secondary vectors and neglected species of mosquitoes are lacking. Cross-species transferability of microsatellite loci between close taxa has been used as a reliable option, being successfully done for Aedes taeniorhynchus Wiedemann (Diptera: Culicidae), Triatoma sordida Stål (Hemiptera: Reduviidae), and Aedes fluviatilis Lutz (Diptera: Culicidae). Nonetheless, prior to the utilization of transferred microsatellites, they must be assessed for their performance and reliability on the target species. Therefore, the goal of this study was to transfer and validate SSR primers previously developed for the Culex pipiens complex to Culex (Culex) nigripalpus Theobald (Diptera: Culicidae). Culex nigripalpus were collected in seven different locations throughout the city of São Paulo using CDC traps baited with dry ice between 2011 and 2013, yielding a total of 223 specimens. Six SSR primers were successfully transferred to Cx. nigripalpus yielding satisfactory results for allele richness, ranging from 5 to 12 alleles per locus. The expected heterozygosity values were higher than the observed heterozygosity (P < 0.05) for the loci CxqCTG10, CxqGT51, and Cxpq59. Linkage disequilibrium results were statistically significant only for the comparison between the loci CxqCA115 and CxqCTG10, and the estimation for the probability of null alleles was also low, ranging from 0 to 0.052.These results substantially validate the SSRs transference to Cx. nigripalpus as well as their subsequent use in microevolution studies comprising this species.

Retention of ancestral polymorphism in Culex nigripalpus (Diptera: Culicidae) from São Paulo, Brazil

Culex nigripalpus Theobald (Diptera: Culicidae) is a native species of Brazil that is well adapted to urban environments and found extensively in the city of São Paulo, Brazil. As a native species, it has been present in this region since long before the foundation of the city, but over time Cx. nigripalpus populations have been affected by man-made changes to the environment. We hypothesize that the populations analyzed in this study constituted a large Cx. nigripalpus population that separated into smaller populations as a result of increased levels of urbanization in the city, and that such high levels of urbanization would result in a genetic homogenization effect. We therefore investigated the microgeographic genetic structure and microevolutionary processes in Cx. nigripalpus populations from seven different locations in the city of São Paulo using a set of six microsatellite primers originally developed for Culex quinquefasciatus and Culex pipiens. Our results indicate that Cx. nigripalpus did not benefit from urbanization and is currently under selective pressures caused by anthropogenic changes and that populations from areas with higher levels of urbanization exhibited similar genetic patterns and low levels of polymorphism, contrasting with the more sylvatic SHA population. These findings are likely to contribute to a better understanding of how anthropogenic selective pressures are driving population genetics and, to some extent, the dynamics of Cx. nigripalpus populations. They should also help elucidate the effects that urbanization processes have on the ecology and behavior of these mosquito populations.