Immature mosquitoes in bamboo internodes in municipal parks, city of são paulo, Brazil.

We conducted an inventory of the mosquito fauna of the internodes of bamboo plants grown in municipal parks in the city of São Paulo, Brazil. During October 2010 to July 2013, a total of 8,541 immature stages belonging to 21 mosquito species were collected from bamboos over 61 municipal parks. Of these, Aedes albopictus was the most abundant species followed by Ae. aegypti found in broken bamboos in most parks. The former species was 7.2 times more abundant than the latter when both species shared the same habitat. Other species collected from bamboos included Culex quinquefasciatus and Haemagogus leucocelaenus. In bamboos with perforated internodes, species of the genus Wyeomyia were the most prevalent. Differences were also observed in species composition and abundance of mosquitoes collected in transversely broken bamboos and those collected from perforated bamboo internodes. Constant surveillance of these breeding sites is crucial due to the epidemiological importance of the species found. Furthermore, these breeding sites may help maintain some native wild mosquito populations along with a variety of other invertebrates found in these urban green areas.

Comparative study of distribution of anopheline vectors (Diptera: Culicidae) in areas with and without malaria transmission in the highlands of an extra-Amazonian region in Brazil.

This study compares the distribution of anopheline mosquitoes in a malaria-endemic municipality (MAL) and a malaria-free municipality (FREE) in an area of the Atlantic Forest in Brazil. Simultaneous quarterly nightly captures were made in three locations in each municipality. One Shannon light trap (Shannon light traps were home made according to specifications published in Am. J. Trop. Med. 1939; 19: 131-140) (SLT) and five CDC light traps (a kind of automatic trap fed by batteries of 12 V and 7 amp/h, with dry ice as a source of CO2; John W. Hock Company, Gainesville, FL) (CLT) (two in the canopy and three at ground level) were operated from 6:00 p.m. to 6:00 a.m. More specimens were captured in MAL (362 in SLTs and 126 in CLTs) than in FREE (66 in SLTs and 59 in CLTs). For the SLTs, Simpson's dominance index was similar in MAL and FREE (D = 0.15 versus D = 0.203, P > 0.7), but Shannon's diversity index was higher for MAL = 1.969 versus H = 1.641, P < 0.01). For the CLTs, Simpson's dominance index was higher in MAL (D = 0.416 versus 0.2688, P < 0.001), and the Shannon diversity index was higher in FREE (H = 1.5222 versus H = 1.115, P < 0.01). In SLTs, Anopheles (Kerteszia) cruzii s.l. frequencies were higher in MAL (chi2 = 23.39; P = 0.000001). In CLTs, An. cruzii s.l. was present in all strata in MAL but only in the canopy inside the forest in FREE (17 specimens). An. cruzii s.l. represented a higher proportion of anophelines in MAL (chi2 = 31; P < 0.000001). The factors that differed in these two areas were anopheline species density and An. cruzii s.l. abundance and distribution.

Characterization of insect-specific Culex flavivirus (Flaviviridae) nucleotide sequences in mosquitoes from urban parks in São Paulo, Brazil.

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.

Influence of water's physical and chemical parameters on mosquito (Diptera: Culicidae) assemblages in larval habitats in urban parks of São Paulo, Brazil.

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.

Kerteszia subgenus of Anopheles associated with the Brazilian Atlantic rainforest:current knowledge and future challenges.

BACKGROUND: The Atlantic rainforest ecosystem, where bromeliads are abundant, provides an excellent environment for Kerteszia species, because these anophelines use the axils of those plants as larval habitat. Anopheles (K.) cruzii and Anopheles (K.) bellator are considered the primary vectors of malaria in the Atlantic forest. Although the incidence of malaria has declined in some areas of the Atlantic forest, autochthonous cases are still registered every year, with Anopheles cruzii being considered to be a primary vector of both human and simian Plasmodium. METHODS: Recent publications that addressed ecological aspects that are important for understanding the involvement of Kerteszia species in the epidemiology of malaria in the Atlantic rainforest in the Neotropical Region were analysed. CONCLUSION: The current state of knowledge about Kerteszia species in relation to the Atlantic rainforest ecosystem was discussed. Emphasis was placed on ecological characteristics related to epidemiological aspects of this group of mosquitoes. The main objective was to investigate biological aspects of the species that should be given priority in future studies.

First record of Anopheles (Anopheles) costai Fonseca & Ramos, 1939 in Espírito Santo State, Brazil.

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.

[Aedes albopictus seasonality in a protected area in the city of São Paulo, Brazil]. / Sazonalidade de Aedes albopictus em área protegida na cidade de São Paulo, Brasil.

The study had the objective of assessing the seasonality of Aedes albopictus and the impact of environmental factors on breeding sites in a protected urban area. Immature individuals were collected through fluid aspiration from nine tree holes, in the Ecological Park of Tietê in the city of São Paulo, Southern Brazil, from 2001 to 2002. The index of positivity and number of individuals results were as follows: positivity x precipitation (rho=0.69, p<0.001), positivity x temperature (rho=0.35, p<0.001), number of individuals x precipitation (rho=0.29, p<0.001) and number of individuals x temperature (rho=0.13, p<0.05). These correlations suggest rainfalls had greater impact than temperature.

[Richness and abundance of Culicidae (Diptera) in an area impacted, Mato Grosso do Sul, Midwestern Brazil]. / Riqueza e abundância de Culicidae (Diptera) em área impactada, Mato Grosso do Sul, Brasil.

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.

[Capture of culicids in urban areas: evaluation of the resting box method]. / Captura de culicídeos em área urbana: avaliação do método das caixas de repouso.

OBJECTIVE: To evaluate the occurrence of adult culicid populations in urban areas and measure the sensitivity of the resting box collection method. METHODS: Mosquito were collected in 1999 and 2000, in two cities in the State of São Paulo, Brazil: Ocauçu and Uchoa. In each city, 15 blocks were drawn by lots and then one home in each block was drawn. Two resting boxes were installed at each home: one inside and the other outside the house. Monthly collections were carried out at each home, over a 13-month period, using manual aspirators inside and outside the home and inside the boxes. The captured specimens were taken to the laboratory for screening and identification according to species and sex. RESULTS: Out of the 2,112 culicid specimens collected, 99.7% were of four species: Culex quinquefasciatus, Aedes aegypti, Cx. declarator and Cx. coronator. The distribution of these species in Ocauçu was 83.3%, 3.2%, 10.8% and 24%, respectively, and in Uchoa it was 83.8%, 8.4%, 4.4% and 3.0%, respectively. Among the females of the genus Culex, 34.3% were captured in the resting boxes and 59.9% were collected from inside the house. Among the females of Ae. aegypti, 17.6% were found in the resting boxes and 82.4% inside the home. CONCLUSIONS: The great majority of the specimens collected belonged to four species of culicids, and Cx. quinquefasciatus was the most common. Proportionally, the females of Ae. aegypti were found more inside the home than were those of the genus Culex. Resting boxes present potential for use as surveillance devices, but their use needs to be more thoroughly evaluated.

Mosquitoes of the Caatinga: 2. Species from periodic sampling of bromeliads and tree holes in a dry Brazilian forest.

The Caatinga is a dry tropical forest, located in the Brazilian semiarid region and rich in phytotelmata. This study investigated the culicid fauna of phytotelmata of the caatinga by sampling for 19 consecutive months aquatic immatures from tree holes and bromeliads. A total of 127L of water was taken from the plants, containing 6764 immature culicids of 16 species, of which 11 (69%) are undescribed and respond to 90% of the total abundance of the specimens collected. Epiphytic bromeliads harbor a large number of immature Culicidae, although terrestrial bromeliads are the most abundant and widely distributed in the region. The richness of culicid species was similar between terrestrial and epiphytic bromeliads and lower in habitats represented by tree hole phytotelmata. There was no similarity in the composition of culicid species that developed in bromeliads or tree holes. Temperature and humidity were the environmental parameters most strongly associated with the proportion of positive plants. The Caatinga has a great number of endemic species that remain unknown to science and many additional culicid species may await discovery from there.

Mosquitoes of the Caatinga: 1. Adults stage survey and the emerge of seven news species endemic of a dry tropical forest in Brazil.

The Caatinga is the least known Brazilian biome in terms of the diversity of Culicidae. No systematic study of the diversity or ecology of the mosquitoes of this biome is available, despite the importance of vector diseases in Brazil. The present study addressed the mosquito biodiversity in the Caatinga biome by sampling adult populations. Specimens were sampled monthly from March 2013 to September 2014 in a Caatinga conservation unit located in the Brazilian semiarid zone. Mosquito collections were carried out in Shannon traps from late afternoon to early evening, and manual aspiration was used to capture diurnal species as well. A total of 4,692 mosquitoes were collected. The most dominant and constant species were all undescribed species belonging to the genera Wyeomyia and Runchomyia, which together represented 80% of the specimens. The most abundant species of epidemiological importance was Haemagogus (Con.) leucocelaenus. The abundance of mosquitoes was positively associated with the relative humidity and temperature recorded during the month preceding the collection date. In the Caatinga, the diversity of adult mosquitoes was associated with the availability (quantity and diversity) of natural larval habitats found in the different phytophysiognomies of the biome, which vary according to temperature and humidity. The number of species unknown to science reflects the levels of endemism that exist in the study area, and reinforces the need to further taxonomic investigation in the biome.

Characterization of insect-specific Culex flavivirus (Flaviviridae) nucleotide sequences in mosquitoes from urban parks in São Paulo, Brazil

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.

Assessment of the correlation between wing size and body weight in captive Culex quinquefasciatus.

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.

Detection of Culex flavivirus and Aedes flavivirus nucleotide sequences in mosquitoes from parks in the city of São Paulo, Brazil.

The dengue viruses are widespread in Brazil and are a major public health concern. Other flaviviruses also cause diseases in humans, although on a smaller scale. The city of São Paulo is in a highly urbanized area with few green spaces apart from its parks, which are used for recreation and where potential vertebrate hosts and mosquito vectors of pathogenic Flavivirus species can be found. Although this scenario can contribute to the transmission of Flavivirus to humans, little is known about the circulation of members of this genus in these areas. In light of this, the present study sought to identify Flavivirus infection in mosquitoes (Diptera: Culicidae) collected in parks in the city of São Paulo. Seven parks in different sectors of the city were selected. Monthly mosquito collections were carried out in each park from March 2011 to February 2012 using aspiration and traps (Shannon and CD C-CO2). Nucleic acids were extracted from the mosquitoes collected and used for reverse-transcriptase and real-time polymerase chain reactions with genus-specific primers targeting a 200-nucleotide region in the Flavivirus NS5 gene. Positive samples were sequenced, and phylogenetic analyses were performed. Culex and Aedes were the most frequent genera of Culicidae collected. Culex flavivirus (CxFV)-related and Aedes flavivirus (AEFV)- related nucleotide sequences were detected in 17 pools of Culex and two pools of Aedes mosquitoes, respectively, among the 818 pools of non-engorged females analyzed. To the best of our knowledge, this is the first report of CxFV and AEFV in the city of São Paulo and Latin America, respectively. Both viruses are insect- specific flaviviruses, a group known to replicate only in mosquito cells and induce a cytopathic effect in some situations. Hence, our data suggests that CxFV and AEFV are present in Culex and Aedes mosquitoes, respectively, in parks in the city of São Paulo. Even though Flavivirus species of medical importance were not detected, surveillance is recommended in the study areas because of the presence of vertebrates and mosquitoes that could act as amplifying hosts and vectors of flaviviruses, providing the required conditions for circulation of these viruses.

[Culicidae mosquito abundance in a preserved metropolitan area and its epidemiological implications]. / Abundância de culicídeos em área metropolitana preservada e suas implicações epidemiológicas.

OBJECTIVE: To study the adult Culicidae fauna in a preserved area in urban outskirts for describing species of public health concern. METHODS: This study was developed in the Tiet Ecological Park located in the outskirt of the city of S o Paulo, Brazil. Monthly collections were undertaken in three habitats at different times using electric aspirators and CDC and Shannon traps from August 1996 to March 1998. Data analysis was based on estimates of frequencies and abundance. RESULTS: Twenty-five species or generic groups were identified among 53,496 specimens collected. Ochlerotatus scapularis, Culex quinquefasciatus, and Culex declarator were the most frequent and abundant species. CONCLUSIONS: The most frequent and abundant species have been implicated in disease transmission to man in other regions. Attention must be given to these Culicidae since the study area is located near to an urban center, which could amplify the conditions for spreading vector diseases.

[Africanized bees in a metropolitan area of Brazil: shelters and climatic influences]. / Abelhas africanizadas em área metropolitana do Brasil: abrigos e influências climáticas.

OBJECTIVE: Africanized bees are the most aggressive bees, form swarms several times a year and build their nests in a great variety of places different from European bees. This behavior brings them into closer contact with human populations, thus giving rise to a larger number of injuries. A study was carried out to gain greater knowledge on these bees' behavior and thus create more efficient strategies for dealing with them and educating the population on this regard. METHODS: Data consisted of 3,061 registries of requests to the Center for the Control of Animal Diseases of S o Paulo for removing bee colonies and wandering swarms in the period between 1994 and 1997. An evaluation of the most frequent nesting sites and their correlation with climatic variables was conducted using Pearson's coefficient. RESULTS: There were 3,061 requests in the study period and the daily volume had a positive correlation with average temperature and degree of insolation and a negative correlation with relative humidity and rainfall. The colonies showed a likelihood for nesting in artificial constructions and the wandering swarms for nesting in trees. CONCLUSIONS: Those periods of the year characterized by high temperatures and low rainfall are related to greater activity of bees and a larger number of wandering swarms, thus giving rise to increased contact with human populations. Items such as boxes and oil-drums should not be left outdoors and ceilings and walls should be sealed off, as they are potential nesting sites for colonies and swarms. Wooded areas may harbor swarms, thus care should always be taken when in the proximity of such areas. No attempt should be made to handle swarms.

Malaria outside the Amazon region: natural Plasmodium infection in anophelines collected near an indigenous village in the Vale do Rio Branco, Itanhaém, SP, Brazil.

A few cases of Plasmodium vivax malaria in which anophelines of subgenus Kerteszia were incriminated as vectors have been reported outside the Amazon region, in the Atlantic Forest. This study was carried out near an indigenous Guarani village in the Curucutu reserve, an environmental protection area in the municipality of Itanhaém in the state of São Paulo, Brazil, on November 30, 2009, February 18, 2010, April 29, 2010 and May 26, 2010. Mosquitoes were collected along the route to the Guarani village where the edge of the Branco river floodplain meets the forests on the mountain slopes. Adult forms were collected with CO(2)-baited CDC traps and Shannon traps from twilight to 10:00 P.M. Anopheles cruzii predominated in both traps. The other species collected in the CDC traps were An. pseudomaculipes/maculipes, An. fluminensis and An. mediopunctatus/forattinii/costai. In addition to the latter three species, An. apicimacula/intermedius and An. strodei were also found in the Shannon traps. All but An. cruzii and An. strodei belong to subgenus Anopheles. A total of 506 mosquitoes were assayed by PCR to detect natural infection by Plasmodium species. In the CDC traps, An. fluminensis and An. pseudomaculipes/maculipes were positive for Plasmodium malariae, while in the Shannon traps An. pseudomaculipes/maculipes was positive for Plasmodium vivax and Plasmodium malariae and An. cruzii was positive for P. malariae, resulting in a minimum infection rate of 0.24%. Our findings suggest that An. cruzii may be incriminated in the transmission of malaria between monkeys and humans, as this species was found to be infected by P. malariae. They also highlight the need for an understanding of the role of anophelines from outside subgenus Kerteszia in the transmission of malaria in the Atlantic Forest, as these were also found to be naturally infected by P. vivax and P. malariae.

Natural infection in anopheline species and its implications for autochthonous malaria in the Atlantic Forest in Brazil.

BACKGROUND: A descriptive study was carried out in an area of the Atlantic Forest with autochthonous malaria in the Parelheiros subdistrict on the periphery of the municipality of São Paulo to identify anopheline fauna and anophelines naturally infected with Plasmodium as well as to discuss their role in this peculiar epidemiological context. METHODS: Entomological captures were made from May 2009 to April 2011 using Shannon traps and automatic CDC traps in four areas chosen for their different patterns of human presence and incidences of malaria (anthropic zone 1, anthropic zone 2, transition zone and sylvatic zone). Natural Plasmodium infection was detected by nested PCR based on amplification of the 18S rRNA gene. RESULTS: In total, 6,073 anophelines were collected from May 2009 to April 2011, and six species were identified in the four zones. Anopheles cruzii was the predominant species in the three environments but was more abundant in the sylvatic zone. Anopheles (Kerteszia) cruzii specimens from the anthropic and sylvatic zones were positive for P. vivax and P. malariae. An. (Ker.) bellator, An. (Nys.) triannulatus, An. (Nys.) strodei, An. (Nys.) lutzi and An. (Ano) maculipes were found in small numbers. Of these, An. (Nys.) triannulatus and An. (Nys.) lutzi, which were collected in the anthropic zone, were naturally infected with P. vivax while An. (Nys.) triannulatus from the anthropic zones and An. (Nys.) strodei from the transition zone were positive for P. malariae. CONCLUSION: These results confirm that Anopheles (Kerteszia) cruzii plays an important role as a major Plasmodium vector. However, the finding of other naturally infected species may indicate that secondary vectors are also involved in the transmission of malaria in the study areas. These findings can be expected to help in the implementation of new measures to control autochthonous malaria in areas of the Atlantic Forest.

Effects of Artificial Flooding for Hydroelectric Development on the Population of Mansonia humeralis (Diptera: Culicidae) in the Paraná River, São Paulo, Brazil.

The closure of two phases of the dam at the Porto Primavera Hydroelectric Plant on the Paraná River flooded a flawed system located in the Municipality of Presidente Epitácio, São Paulo state, favoring the proliferation of aquatic weeds. This study aimed to observe the population of Mansonia humeralis in the area, monitoring the richness, diversity, and dominance of this species both before and during different phases of reservoir flooding as well as evaluate its possible consequences concerning human and animal contact. Adult mosquitoes were collected monthly in the following periods: at the original level, after the first flood, and after the maximum level had been reached between 1997 and 2002. Collection methods used were an aspirator, a Shannon trap, and the Human Attractive Technique. A total of 30,723 mosquitoes were collected, Ma. humeralis accounting for 3.1% in the preflood phase, 59.6% in the intermediate, and 53.8% at maximum level. This species is relevant to public health, since the prospect of continued contact between Ma. humeralis and the human population enhances the dam's importance in the production of nuisance mosquitoes, possibly facilitating the transmission of arboviruses. Local authorities should continue to monitor culicid activity through sustainable entomological surveillance.