Genetic Structuring of One of the Main Vectors of Sylvatic Yellow Fever: Haemagogus (Conopostegus) leucocelaenus (Diptera: Culicidae).

Genetic diversity and population structuring for the species Haemogogus leucocelaenus, a sylvatic vector of yellow fever virus, were found to vary with the degree of agricultural land use and isolation of fragments of Atlantic Forest in municipalities in the state of São Paulo where specimens were collected. Genotyping of 115 mitochondrial SNPs showed that the populations with the highest indices of genetic diversity (polymorphic loci and mean pairwise differences between the sequences) are found in areas with high levels of agricultural land use (northeast of the State). Most populations exhibited statistically significant negative values for the Tajima D and Fu FS neutrality tests, suggesting recent expansion. The results show an association between genetic diversity in this species and the degree of agricultural land use in the sampled sites, as well as signs of population expansion of this species in most areas, particularly those with the highest forest edge densities. A clear association between population structuring and the distance between the sampled fragments (isolation by distance) was observed: samples from a large fragment of Atlantic Forest extending along the coast of the state of São Paulo exhibited greater similarity with each other than with populations in the northwest of the state.

The role of forest fragmentation in yellow fever virus dispersal.

The intense process of deforestation in tropical forests poses serious challenges for the survival of biodiversity, as well as for the human species itself. This scenario is supported by the increase in the incidence of epidemics of zoonotic origin observed over the last few decades. In the specific case of sylvatic yellow fever (YF), it has already been shown that an increase in the transmission risk of the causative agent (yellow fever virus - YFV) is associated with areas with a high degree of forest fragmentation, which can facilitate the spread of the virus. In this study we tested the hypothesis that areas with more fragmented landscapes and a higher edge density (ED) but a high degree of connectivity between forest patches favor YFV spread. To this end, we used YF epizootics in non-human primates (NHPs) in the state of São Paulo to build direct networks, and used a multi-selection approach to analyze which landscape features could facilitate YFV spread. Our results showed that municipalities with the potential to spread the virus exhibited a higher amount of forest edge. Additionally, the models with greater empirical support showed a strong association between forest edge density and the risk of occurrence of epizootic diseases, as well as the need for a minimum threshold of native vegetation cover to restrict their transmission. These findings corroborate our hypothesis that more fragmented landscapes with a higher degree of connectivity favor the spread of YFV, while landscapes with fewer connections tend to act as dead zones for the circulation of the virus.

Evaluation of Methods for Collecting Mosquitoes (Culicidae: Diptera) in Canopy and Ground Strata in the Brazilian Savanna.

The hand-net is the standard method for capturing mosquitoes with sylvatic diurnal activity in disease outbreaks in Brazil. However, occupational risks and biases related to the collectors' abilities and attractiveness are important limitations. In this study, we compared hand-nets with automatic traps (CDC) associated to CO2 and BG-Lure® in the Vassununga State Park, a Brazilian Savanna protection area. The collections carried out over 27 days on the ground and the forest canopy. A total of 1555 mosquitoes were obtained in 20 taxa. The diversity index ranged between 1.12 and 1.79 and the dominance index from 0.22 to 0.40. The dominant species on the ground was Aedes scapularis (46.0%), and in the canopy, Hg. janthinomys/capricornii (31.9%). Haemagogus leucocelaenus was rare (n = 2). The hand-net resulted in the greatest diversity and abundance of species in both strata, followed by the traps associated with CO2. A low degree of similarity was observed between the hand-net on the ground compared to the other capture methods. The use of BG-Lure® alone resulted in a low number of specimens. In conclusion, the hand-net is still the method of choice for collecting arbovirus vectors in the diurnal period, especially yellow fever vectors.

Evaluation of Methods to Collect Diurnal Culicidae (Diptera) at Canopy and Ground Strata, in the Atlantic Forest Biome.

Hand-held insect nets are the standard method for capturing vector mosquitoes of sylvatic arboviruses; however, occupational risks and biases due to individual skill and attractiveness are important limitations. The use of chemical attractants and automatic traps could be an alternative to resolve these limitations. This study compares the yields achieved using nets with those employing electrical traps with CO2 and BG-Lure®, near the ground and in the canopy strata (6.0 and 8.0 m high). The study was conducted at the Cantareira State Park, which is in the Brazilian Atlantic Forest biome. In the 18 collections performed, 3570 specimens of 52 taxa were obtained. The most frequent species captured near the ground were Wyeomyia confusa and Limatus durhamii, whereas Sabethes albiprivus, Sabethes purpureus, and Haemagogus leucocelaenus were the most frequent in the canopy. The nets resulted in greater species richness and abundance, followed by the trap employing CO2. The combination of CO2 traps with BG-Lure® did not improve performance. The use of BG-Lure® alone resulted in low abundance and a low number of species. Our results demonstrate that the use of traps with CO2 can be complementary to collections with nets; however, for species of epidemiological interest such as those of the genera Haemagogus and Sabethes, especially in the canopy, the net remains the method of choice.

The influence of landscape structure on the dispersal pattern of yellow fever virus in the state of São Paulo.

Man-made changes to the landscape play a crucial role in altering the epidemiologic patterns of infectious diseases, mainly as a result of pathogen spillover. Sylvatic yellow fever is ideally suited to modeling of this phenomenon as the risk of transmission of the disease as well as its circulation and dispersal are associated with forest fragmentation. In this study we investigated the temporal dispersal pattern of yellow fever virus (YFV) by means of confirmed cases of epizootics in non-human primates in municipalities in the state of São Paulo where there was no recommendation for vaccination in 2017. We analyzed the resistance to dispersal associated with different classes of land use and the geographic distances between the different locations where epizootics were recorded. The model that best explained the temporal dispersal pattern of YFV in the study area indicated that this was influenced by the geographic distance between collection locations and by the permeability of the forest edges (150 m) at the interface with the following core areas: Water, Agricultural, Non-Forest Formation and Forestry. Water, Agricultural, Urban and Forest core areas and the interfaces between the latter two formed important barriers to circulation of the virus. These findings indicate that fragmentation of vegetation tends to decrease the time taken for pathogens to spread, while conservation of forest areas has the opposite effect.

The Influence of the pH and Salinity of Water in Breeding Sites on the Occurrence and Community Composition of Immature Mosquitoes in the Green Belt of the City of São Paulo, Brazil.

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.

Relationship between vertical stratification and feeding habits of mosquito (Diptera: Culicidae) assemblages collected in conservation units in the green belt of the city of São Paulo, Brazil.

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.

A mathematical model for zoonotic transmission of malaria in the Atlantic Forest: Exploring the effects of variations in vector abundance and acrodendrophily.

Transmission foci of autochthonous malaria caused by Plasmodium vivax-like parasites have frequently been reported in the Atlantic Forest in Southeastern and Southern Brazil. Evidence suggests that malaria is a zoonosis in these areas as human infections by simian Plasmodium species have been detected, and the main vector of malaria in the Atlantic Forest, Anopheles (Kerteszia) cruzii, can blood feed on human and simian hosts. In view of the lack of models that seek to predict the dynamics of zoonotic transmission in this part of the Atlantic Forest, the present study proposes a new deterministic mathematical model that includes a transmission compartment for non-human primates and parameters that take into account vector displacement between the upper and lower forest strata. The effects of variations in the abundance and acrodendrophily of An. cruzii on the prevalence of infected humans in the study area and the basic reproduction number (R0) for malaria were analyzed. The model parameters are based on the literature and fitting of the empirical data. Simulations performed with the model indicate that (1) an increase in the abundance of the vector in relation to the total number of blood-seeking mosquitoes leads to an asymptotic increase in both the proportion of infected individuals at steady state and R0; (2) the proportion of infected humans at steady state is higher when displacement of the vector mosquito between the forest strata increases; and (3) in most scenarios, Plasmodium transmission cannot be sustained only between mosquitoes and humans, which implies that non-human primates play an important role in maintaining the transmission cycle. The proposed model contributes to a better understanding of the dynamics of malaria transmission in the Atlantic Forest.

Complexity of malaria transmission dynamics in the Brazilian Atlantic Forest.

Plasmodium malariae and Plasmodium vivax are protozoan parasites that can cause malaria in humans. They are genetically indistinguishable from, respectively, Plasmodium brasilianum and Plasmodium simium, i.e. parasites infecting New World non-human primates in South America. In the tropical rainforests of the Brazilian Atlantic coast, it has long been hypothesized that P. brasilianum and P. simium in platyrrhine primates originated from P. malariae and P. vivax in humans. A recent hypothesis proposed the inclusion of Plasmodium falciparum into the transmission dynamics between humans and non-human primates in the Brazilian Atlantic tropical rainforest. Herein, we assess the occurrence of human malaria in simians and sylvatic anophelines using field-collected samples in the Capivari-Monos Environmental Protection Area from 2015 to 2017. We first tested simian blood and anopheline samples. Two simian (Aloutta) blood samples (18%, n = 11) showed Plasmodium cytb DNA sequences, one for P. vivax and another for P. malariae. From a total of 9,416 anopheline females, we found 17 pools positive for Plasmodium species with a 18S qPCR assay. Only three showed P. cytb DNA sequence, one for P. vivax and the others for rodent malaria species (similar to Plasmodium chabaudi and Plasmodium berghei). Based on these results, we tested 25 rodent liver samples for the presence of Plasmodium and obtained P. falciparum cytb DNA sequence in a rodent (Oligoryzomys sp.) liver. The findings of this study indicate complex malaria transmission dynamics composed by parallel spillover-spillback of human malaria parasites, i.e. P. malariae, P. vivax, and P. falciparum, in the Brazilian Atlantic forest.

Detection of Zika and dengue viruses in wild-caught mosquitoes collected during field surveillance in an environmental protection area in São Paulo, Brazil.

Species of the genus Flavivirus are widespread in Brazil and are a major public health concern. The country's largest city, São Paulo, is in a highly urbanized area with a few forest fragments which are commonly used for recreation. These can be considered to present a potential risk of flavivirus transmission to humans as they are home simultaneously to vertebrate hosts and mosquitoes that are potential flavivirus vectors. The aim of this study was to conduct flavivirus surveillance in field-collected mosquitoes in the Capivari-Monos Environmental Protection Area (EPA) and identify the flavivirus species by sequence analysis in flavivirus IFA-positive pools. Monthly mosquito collections were carried out from March 2016 to April 2017 with CO2-baited CDC light traps. Specimens were identified morphologically and grouped in pools of up to 10 individuals according to their taxonomic category. A total of 260 pools of non-engorged females were inoculated into C6/36 cell culture, and the cell suspensions were analyzed by indirect immunofluorescence assay (IFA) after the incubation period. IFA-positive pools were tested by qRT-PCR with genus-specific primers targeting the flavivirus NS5 gene to confirm IFA-positive results and sequenced to identify the species. Anopheles cruzii (19.5%) and Wyeomyia confusa (15.3%) were the most frequent vector species collected. IFA was positive for flaviviruses in 2.3% (6/260) of the sample pools. This was confirmed by qRT-PCR in five pools (83.3%). All five flavivirus-positive pools were successfully sequenced and the species identified. DENV serotype 2 (DENV-2) was detected in Culex spp. and Culex vaxus pools, while ZIKV was identified in An. cruzii, Limatus durhamii and Wy. confusa pools. To the best of our knowledge, detection of flavivirus species of medical importance has never previously been reported in these species of wild-caught mosquitoes. The finding of DENV-2 and ZIKV circulating in wild mosquitoes suggests the existence of an enzootic cycle in the area. In-depth studies of DENV-2 and ZIKV, including investigation of mosquito infection, vector competence and infection in sylvatic hosts, are needed to shed light on the transmission dynamics of these important viruses and the potential risk of future outbreaks of DENV-2 and ZIKV infections in the region.

Possible non-sylvatic transmission of yellow fever between non-human primates in São Paulo city, Brazil, 2017-2018.

Yellow Fever (YF) is a severe disease caused by Yellow Fever Virus (YFV), endemic in some parts of Africa and America. In Brazil, YFV is maintained by a sylvatic transmission cycle involving non-human primates (NHP) and forest canopy-dwelling mosquitoes, mainly Haemagogus-spp and Sabethes-spp. Beginning in 2016, Brazil faced one of the largest Yellow Fever (YF) outbreaks in recent decades, mainly in the southeastern region. In São Paulo city, YFV was detected in October 2017 in Aloutta monkeys in an Atlantic Forest area. From 542 NHP, a total of 162 NHP were YFV positive by RT-qPCR and/or immunohistochemistry, being 22 Callithrix-spp. most from urban areas. Entomological collections executed did not detect the presence of strictly sylvatic mosquitoes. Three mosquito pools were positive for YFV, 2 Haemagogus leucocelaenus, and 1 Aedes scapularis. In summary, YFV in the São Paulo urban area was detected mainly in resident marmosets, and synanthropic mosquitoes were likely involved in viral transmission.

Diversity analysis and an updated list of mosquitoes (Diptera: Culicidae) found in Cantareira State Park, São Paulo, Brazil.

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.

Importance of microenvironment to arbovirus vector distribution in an urban area, São Paulo, Brazil.

INTRODUCTION: The study of the landscape ecology, biological microhabitat, and epidemiological implications for the distribution of the main vectors Aedes aegypti and Aedes albopictus contribute to the prevention and control actions regarding the diseases they transmit. METHODS: This study sought to assess data on positive properties of the vector control program activities from 1998 to 2010. An entomological survey was also carried out on a sample of buildings collecting larvae and pupae from containers between October and April (spring / summer) from 2002 to 2005. We assessed the physico-chemical data of the water in 20% of positive containers. The vegetation and urbanization were assessed with the aid of satellite images and microenvironments were classified as urbanized, woods, and shrubs. The data were analyzed using statistical and geoprocessing software. RESULTS: Ae. aegypti and Ae. albopictus colonized all types of microhabitats and microenvironments, predominantly in the urbanized area, in isolation and in coexistence. The microhabitat of Ae. aegypti showed a temperature gradient greater than that of Ae. albopictus, and there was an association with urbanized areas for the first species and wooded areas for the last species. CONCLUSIONS: Landscape ecology and intra-urban differences favor different microclimates, which contribute to the coexistence of species in the urban environment in an area close to the forest, raising the risk of other arbovirus infections in urban areas. The ecological niche should be considered for Ae. albopictus. Entomological and virologic monitoring are suggested as arbovirus surveillance actions in urban infested centers near preserved forests.

Genomic evidence of yellow fever virus in Aedes scapularis, southeastern Brazil, 2016.

The southeastern region of Brazil has recently experienced the largest yellow fever disease outbreak in decades. Since July 2016 epizootic events were reported in São Paulo state's north region, where 787 Culicidae were captured as part of public health surveillance efforts and tested using real-time quantitative PCR. One Aedes scapularis pool collected in November 2016 in an agriculture area in Urupês city tested positive for YFV-RNA. Using a validated multiplex PCR approach we were able to recover a complete virus genome sequence from this pool. Phylogenetic analysis of the novel strain and publicly available data indicates that the belongs to the South American genotype 1 clade circulating in Sao Paulo state and is basal to the recent outbreak clade in southeast Brazil. Our findings highlight the need of additional studies, including vector competence studies, to disentangle the role of Aedes scapularis in yellow fever transmission in the Americas.

Influence of landscape composition and configuration on the richness and abundance of potential sylvatic yellow fever vectors in a remnant of Atlantic Forest in the city of São Paulo, Brazil.

The landscape's structure can play a relevant role in epidemic patterns of arboviruses, influencing factors such as abundance, movement, and dispersal ability in arthropod vectors and vertebrate hosts, besides promoting alterations in the rate of potential infectious contacts between these organisms. In the Americas, yellow fever (YF) exhibits only the sylvatic cycle, in which the virus circulates in sylvatic areas among non-human primates, being transmitted by mosquitoes of the Haemagogus and Sabethes genera. In this study, we investigate some aspects of the landscape in relation to diversity and abundance of culicid species associated with YF transmission. Studies were performed in the Cantareira State Park, a remnant of the Atlantic Forest located in Greater Metropolitan São Paulo, Brazil, where the YF virus circulated recently with dozens of deaths in howler monkeys (Alouatta guariba), in addition to reported human cases. Mosquito collections were carried out monthly from February 2015 to April 2017. Mosquitoes were collected from three sites using battery-powered aspirator (12-volt battery), CDC, and Shannon traps for adults, and suction samplers and entomological spoons in breeding sites to collect immature forms. 703 mosquitoes belonging to 12 species of the Aedini and Sabethini tribes were collected. Aedes scapularis and Psorophora ferox exhibited higher abundance, while Haemagogus leucocelaenus, the main vector of YF in São Paulo state, showed lower abundance in all sampled areas. The site with longer edge between forest area and anthropic area presented more richness and abundance of YF vector species, while the site with larger forest cover area and shorter edges between forest and anthropic areas exhibited an inverse pattern. Statistically significant differences were observed between the composition of potential YF vector species among the investigated sites. Although Hg. leucocelaenus occurred in all sampled sites, the different patterns of distribution and abundance of other mosquitoes such as Aedes scapularis and Psorophora ferox suggest that these species may be involved in the transmission of sylvatic YF in the study area.

Importance of microenvironment to arbovirus vector distribution in an urban area, São Paulo, Brazil

Abstract INTRODUCTION: The study of the landscape ecology, biological microhabitat, and epidemiological implications for the distribution of the main vectors Aedes aegypti and Aedes albopictus contribute to the prevention and control actions regarding the diseases they transmit. METHODS: This study sought to assess data on positive properties of the vector control program activities from 1998 to 2010. An entomological survey was also carried out on a sample of buildings collecting larvae and pupae from containers between October and April (spring / summer) from 2002 to 2005. We assessed the physico-chemical data of the water in 20% of positive containers. The vegetation and urbanization were assessed with the aid of satellite images and microenvironments were classified as urbanized, woods, and shrubs. The data were analyzed using statistical and geoprocessing software. RESULTS: Ae. aegypti and Ae. albopictus colonized all types of microhabitats and microenvironments, predominantly in the urbanized area, in isolation and in coexistence. The microhabitat of Ae. aegypti showed a temperature gradient greater than that of Ae. albopictus, and there was an association with urbanized areas for the first species and wooded areas for the last species. CONCLUSIONS: Landscape ecology and intra-urban differences favor different microclimates, which contribute to the coexistence of species in the urban environment in an area close to the forest, raising the risk of other arbovirus infections in urban areas. The ecological niche should be considered for Ae. albopictus. Entomological and virologic monitoring are suggested as arbovirus surveillance actions in urban infested centers near preserved forests.

Effects of anthropogenic landscape changes on the abundance and acrodendrophily of Anopheles (Kerteszia) cruzii, the main vector of malaria parasites in the Atlantic Forest in Brazil.

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.

Association of the occurrence of Brazilian spotted fever and Atlantic rain forest fragmentation in the São Paulo metropolitan region, Brazil.

Brazilian Spotted Fever (BSF) is a zoonotic disease caused by the bacterium Rickettsia rickettsii. In the São Paulo Metropolitan Region (SPMR) it is transmitted by Amblyomma aureolatum ticks. In this region, annual lethality of the disease can reach 80% and spatial occurrence depends on environmental factors and more particularly on the presence and interaction of domestic and wild carnivores as well as the presence and characteristics of the remnant Atlantic Rain Forest patches. This study analyzed the association between forest fragmentation and its influence on the risk of occurrence of the disease in the human population. Domestic dogs tested for R. rickettsii antibodies in nine different areas under the influence of different patterns of Rain Forest fragmented landscapes and human occupancy. Landscape metrics were obtained by analyzing satellite images and high-resolution orthophotos. Principal component analysis (PCA) was used to determine among the different landscape variables the one that could best explain the data variance, and the results were tested against canine seroprevalence in order to address disease occurrence risk levels. From 270 canine samples, the seroprevalence ranged from 0 to 37%. PCA showed an inverse correlation between functionally connected large forest patches and the canine seroprevalence for R. rickettsii (p=0.030; Spearman's R=-0.683), while there was a positive correlation between forest border effect and canine seroprevalence (p=0.037; Spearman's R=- 0.909). The further attributed disease occurrence risk level supported the real spatial prevalence of the disease reported for the last eight years (p=0.023; Spearman's R=0.63). The results suggest an important relation of deforestation and fragmentation with the occurrence of BSF in the SPMR.

Vector aspects in risk areas for sylvatic yellow fever in the state of Mato Grosso do Sul, Brazil

Epizootics and yellow fever epidemics in 2008 in Brazil, Paraguay and Argentina marked a significant progression of the disease further south in the continent affecting non-vaccinated human populations. The state of Mato Grosso do Sul is located between the tropical endemic region and the subtropical epidemic region and, despite being an area recommended for vaccination since the 1980s, human and animal cases of the disease are still registered. The purpose of this study was to present the results of entomological surveys of probable sites of infection (PSI) geographically describing these areas regarding human cases and disease suspicion reported in 2008. Thirteen locations in nine municipalities were investigated. A total of 305 females of the genus Haemagogus and Sabethes and five specimens of Aedes albopictus were obtained. The genus Haemagogus was more abundant but Sabethes presented twice as many species (3 x 6) and higher distribution within the collection points. The most abundant species was Hg. janthinomys, with 102 individuals in one location. The other species found were: Hg. leucocelaenus, Hg. spegazzinii, Sa. albiprivus, Sa. belisarioi, Sa. chloropterus, Sa. glaucodaemun, Sa. intermedius and Sa. soperi. It was not possible to identify predominant species in relation to environmental and geographical characteristics of the collection points. Of the nine human cases, eight corresponded to persons not resident in the State, seven of them tourists. MS features a large diversity of rural and wildlife tourist attractions where the yellow fever vectors described in this work are present. In this sense, the monitoring of vaccination coverage and flow of non-vaccinated people through the area are crucial to controlling the disease.