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.

Atlantic Forest Malaria: A Review of More than 20 Years of Epidemiological Investigation.

In the south and southeast regions of Brazil, cases of malaria occur outside the endemic Amazon region near the Atlantic Forest in some coastal states, where Plasmodium vivax is the recognized parasite. Characteristics of cases and vectors, especially Anopheles (Kerteszia) cruzii, raise the hypothesis of a zoonosis with simians as reservoirs. The present review aims to report on investigations of the disease over a 23-year period. Two main sources have provided epidemiological data: the behavior of Anopheles vectors and the genetic and immunological aspects of Plasmodium spp. obtained from humans, Alouatta simians, and Anopheles spp. mosquitoes. Anopheles (K.) cruzii is the most captured species in the forest canopy and is the recognized vector. The similarity between P. vivax and Plasmodium simium and that between Plasmodium malariae and Plasmodium brasilianum shared between simian and human hosts and the involvement of the same vector in the transmission to both hosts suggest interspecies transfer of the parasites. Finally, recent evidence points to the presence of Plasmodium falciparum in a silent cycle, detected only by molecular methods in asymptomatic individuals and An. (K.) cruzii. In the context of malaria elimination, it is paramount to assemble data about transmission in such non-endemic low-incidence areas.

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.

Naturally Acquired Humoral Immunity against Malaria Parasites in Non-Human Primates from the Brazilian Amazon, Cerrado and Atlantic Forest.

Non-human primates (NHPs) have been shown to be infected by parasites of the genus Plasmodium, the etiological agent of malaria in humans, creating potential risks of zoonotic transmission. Plasmodium brasilianum, a parasite species similar to P. malariae of humans, have been described in NHPs from Central and South America, including Brazil. The merozoite surface protein 1 (MSP1), besides being a malaria vaccine candidate, is highly immunogenic. Due to such properties, we tested this protein for the diagnosis of parasite infection. We used recombinant proteins of P. malariae MSP1, as well as of P. falciparum and P. vivax, for the detection of antibodies anti-MSP1 of these parasite species, in the sera of NHPs collected in different regions of Brazil. About 40% of the NHP sera were confirmed as reactive to the proteins of one or more parasite species. A relatively higher number of reactive sera was found in animals from the Atlantic Forest than those from the Amazon region, possibly reflecting the former more intense parasite circulation among NHPs due to their proximity to humans at a higher populational density. The presence of Plasmodium positive NHPs in the surveyed areas, being therefore potential parasite reservoirs, needs to be considered in any malaria surveillance program.

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.

Assessment of asymptomatic Plasmodium spp. infection by detection of parasite DNA in residents of an extra-Amazonian region of Brazil.

BACKGROUND: The hypotheses put forward to explain the malaria transmission cycle in extra-Amazonian Brazil, an area of very low malaria incidence, are based on either a zoonotic scenario involving simian malaria, or a scenario in which asymptomatic carriers play an important role. OBJECTIVES: To determine the incidence of asymptomatic infection by detecting Plasmodium spp. DNA and its role in residual malaria transmission in a non-Amazonian region of Brazil. METHODS: Upon the report of the first malaria case in 2010 in the Atlantic Forest region of the state of Espírito Santo, inhabitants within a 2 km radius were invited to participate in a follow-up study. After providing signed informed consent forms, inhabitants filled out a questionnaire and gave blood samples for PCR, and thick and thin smears. Follow-up visits were performed every 3 months over a 21 month period, when new samples were collected and information was updated. RESULTS: Ninety-two individuals were initially included for follow-up. At the first collection, all of them were clearly asymptomatic. One individual was positive for Plasmodium vivax, one for Plasmodium malariae and one for both P. vivax and P. malariae, corresponding to a prevalence of 3.4% (2.3% for each species). During follow-up, four new PCR-positive cases (two for each species) were recorded, corresponding to an incidence of 2.5 infections per 100 person-years or 1.25 infections per 100 person-years for each species. A mathematical transmission model was applied, using a low frequency of human carriers and the vector density in the region, and calculated based on previous studies in the same locality whose results were subjected to a linear regression. This analysis suggests that the transmission chain is unlikely to be based solely on human carriers, regardless of whether they are symptomatic or not. CONCLUSION: The low incidence of cases and the low frequency of asymptomatic malaria carriers investigated make it unlikely that the transmission chain in the region is based solely on human hosts, as cases are isolated one from another by hundreds of kilometers and frequently by long periods of time, reinforcing instead the hypothesis of zoonotic transmission.

Mitochondrial genome of Plasmodium vivax/simium detected in an endemic region for malaria in the Atlantic Forest of Espírito Santo state, Brazil: do mosquitoes, simians and humans harbour the same parasite?

BACKGROUND: The transmission of malaria in the extra-Amazonian regions of Brazil, although interrupted in the 1960s, has persisted to the present time in some areas of dense Atlantic Forest, with reports of cases characterized by particular transmission cycles and clinical presentations. Bromeliad-malaria, as it is named, is particularly frequent in the state of Espírito Santo, with Plasmodium vivax being the parasite commonly recognized as the aetiologic agent of human infections. With regard to the spatial and temporal distances between cases reported in this region, the transmission cycle does not fit the traditional malaria cycle. The existence of a zoonosis, with infected simians participating in the epidemiology, is therefore hypothesized. In the present study, transmission of bromeliad-malaria in Espírito Santo is investigated, based on the complete mitochondrial genome of DNA extracted from isolates of Plasmodium species, which had infected humans, a simian from the genus Allouata, and Anopheles mosquitoes. Plasmodium vivax/simium was identified in the samples by both nested PCR and real-time PCR. After amplification, the mitochondrial genome was completely sequenced and compared with a haplotype network which included all sequences of P. vivax/simium mitochondrial genomes sampled from humans and simians from all regions in Brazil. RESULTS: The haplotype network indicates that humans and simians from the Atlantic Forest become infected by the same haplotype, but some isolates from humans are not identical to the simian isolate. In addition, the plasmodial DNA extracted from mosquitoes revealed sequences different from those obtained from simians, but similar to two isolates from humans. CONCLUSIONS: These findings strengthen support for the hypothesis that in the Atlantic Forest, and especially in the state with the highest frequency of bromeliad-malaria in Brazil, parasites with similar molecular backgrounds are shared by humans and simians. The recognized identity between P. vivax and P. simium at the species level, the sharing of haplotypes, and the participation of the same vector in transmitting the infection to both host species indicate interspecies transference of the parasites. However, the intensity, frequency and direction of this transfer remain to be clarified.

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.

Circulation of antibodies against yellow fever virus in a simian population in the area of Porto Primavera Hydroelectric Plant, São Paulo, Brazil.

Yellow fever (YF) is an acute viral infectious disease transmitted by mosquitoes which occurs in two distinct epidemiological cycles: sylvatic and urban. In the sylvatic cycle, the virus is maintained by monkey's infection and transovarian transmission in vectors. Surveillance of non-human primates is required for the detection of viral circulation during epizootics, and for the identification of unaffected or transition areas. An ELISA (enzyme-linked immunosorbent assay) was standardized for estimation of the prevalence of IgG antibodies against yellow fever virus in monkey sera (Alouatta caraya) from the reservoir area of Porto Primavera Hydroelectric Plant, in the state of São Paulo, Brazil. A total of 570 monkey sera samples were tested and none was reactive to antibodies against yellow fever virus. The results corroborate the epidemiology of yellow fever in the area. Even though it is considered a transition area, there were no reports to date of epizootics or yellow fever outbreaks in humans. Also, entomological investigations did not detect the presence of vectors of this arbovirus infection. ELISA proved to be fast, sensitive, an adequate assay, and an instrument for active search in the epidemiological surveillance of yellow fever allowing the implementation of prevention actions, even before the occurrence of epizootics.

Circulation of antibodies against yellow fever virus in a simian population in the area of Porto Primavera Hydroelectric Plant, São Paulo, Brazil / Circulação de anticorpos contra o vírus amarílico em população simiana da região da usina hidrelétrica de Porto Primavera, São Paulo, Brasil

Yellow fever (YF) is an acute viral infectious disease transmitted by mosquitoes which occurs in two distinct epidemiological cycles: sylvatic and urban. In the sylvatic cycle, the virus is maintained by monkey's infection and transovarian transmission in vectors. Surveillance of non-human primates is required for the detection of viral circulation during epizootics, and for the identification of unaffected or transition areas. An ELISA (enzyme-linked immunosorbent assay) was standardized for estimation of the prevalence of IgG antibodies against yellow fever virus in monkey sera (Alouatta caraya) from the reservoir area of Porto Primavera Hydroelectric Plant, in the state of São Paulo, Brazil. A total of 570 monkey sera samples were tested and none was reactive to antibodies against yellow fever virus. The results corroborate the epidemiology of yellow fever in the area. Even though it is considered a transition area, there were no reports to date of epizootics or yellow fever outbreaks in humans. Also, entomological investigations did not detect the presence of vectors of this arbovirus infection. ELISA proved to be fast, sensitive, an adequate assay, and an instrument for active search in the epidemiological surveillance of yellow fever allowing the implementation of prevention actions, even before the occurrence of epizootics.

A febre amarela (FA) é doença infecciosa aguda de origem viral transmitida por mosquitos. No ciclo silvestre, o vírus é mantido por meio da infecção de macacos e da transmissão transovariana nos vetores. A vigilância sobre populações de primatas não humanos torna-se necessária para detectar a circulação viral, quando ainda está restrito a epizootias, e para determinar sua presença em regiões indenes ou de transição para a doença. Padronizou-se a técnica ELISA (Enzyme Linked Immunosorbent Assay) para determinar a prevalência de anticorpos da classe IgG contra o vírus da FA em soros de bugios (Alouatta caraya) da região do reservatório da Usina Hidrelétrica de Porto Primavera, SP. Foram testados soros de 570 macacos sendo que nenhuma amostra mostrou-se reativa para a presença de anticorpos contra o vírus da FA. Os resultados são coerentes com a epidemiologia da FA na região. Mesmo sendo área de transição, não se conhece, até o momento, ocorrência de epizootia ou surto de FA em humanos e investigações entomológicas não apontaram a presença de vetores para esta arbovirose. A técnica mostrou-se sensível, rápida e útil à vigilância epidemiológica como instrumento de busca ativa permitindo desencadear ações preventivas, como vacinação, antes mesmo do surgimento de epizootias.

Natural Plasmodium infections in Brazilian wild monkeys: reservoirs for human infections?

Four hundred and forty-eight samples of total blood from wild monkeys living in areas where human autochthonous malaria cases have been reported were screened for the presence of Plasmodium using microscopy and PCR analysis. Samples came from the following distinct ecological areas of Brazil: Atlantic forest (N=140), semideciduous Atlantic forest (N=257) and Cerrado (a savannah-like habitat) (N=51). Thick and thin blood smears of each specimen were examined and Plasmodium infection was screened by multiplex polymerase chain reaction (multiplex PCR). The frequency of Plasmodium infections detected by PCR in Alouatta guariba clamitans in the São Paulo Atlantic forest was 11.3% or 8/71 (5.6% for Plasmodium malariae and 5.6% for Plasmodium vivax) and one specimen was positive for Plasmodium falciparum (1.4%); Callithrix sp. (N=30) and Cebus apella (N=39) specimens were negative by PCR tests. Microscopy analysis was negative for all specimens from the Atlantic forest. The positivity rate for Alouatta caraya from semideciduous Atlantic forest was 6.8% (16/235) in the PCR tests (5.5, 0.8 and 0.4% for P. malariae, P. falciparum and P. vivax, respectively), while C. apella specimens were negative. Parasitological examination of the samples using thick smears revealed Plasmodium sp. infections in only seven specimens, which had few parasites (3.0%). Monkeys from the Cerrado (a savannah-like habitat) (42 specimens of A. caraya, 5 of Callithrix jacchus and 4 of C. apella) were negative in both tests. The parasitological prevalence of P. vivax and P. malariae in wild monkeys from Atlantic forest and semideciduous Atlantic forest and the finding of a positive result for P. falciparum in Alouatta from both types of forest support the hypothesis that monkeys belonging to this genus could be a potential reservoir. Furthermore, these findings raise the question of the relationship between simian and autochthonous human malaria in extra-Amazonian regions.

Natural Plasmodium infections in Brazilian wild monkeys: Reservoirs for human infections?

Four hundred and forty-eight samples of total blood from wild monkeys living in areas where human autochthonous malaria cases have been reported were screened for the presence of Plasmodium using microscopy and PCR analysis. Samples came from the following distinct ecological areas of Brazil: Atlantic forest (N = 140), semideciduous Atlantic forest (N = 257) and Cerrado (a savannah-like habitat) (N = 51). Thick and thin blood smears of each specimen were examined and Plasmodium infection was screened by multiplex polymerase chain reaction (multiplex PCR). The frequency of Plasmodium infections detected by PCR in Alouatta guariba clamitans in the São Paulo Atlantic forest was 11.3% or 8/71 (5.6% for Plasmodium malariae and 5.6% for Plasmodium vivax) and one specimen was positive for Plasmodium falciparum (1.4%); Callithrix sp. (N = 30) and Cebus apella (N = 39) specimens were negative by PCR tests. Microscopy analysis was negative for all specimens from the Atlantic forest. The positivity rate for Alouatta caraya from semideciduous Atlantic forest was 6.8% (16/235) in the PCR tests (5.5, 0.8 and 0.4% for P. malariae, P. falciparum and P. vivax, respectively), while C. apella specimens were negative. Parasitological examination of the samples using thick smears revealed Plasmodium sp. infections in only seven specimens, which had few parasites (3.0%). Monkeys from the Cerrado (a savannah-like habitat) (42 specimens of A. caraya, 5 of Callithrix jacchus and 4 of C. apella) were negative in both tests. The parasitological prevalence of P. vivax and P. malariae in wild monkeys from Atlantic forest and semideciduous Atlantic forest and the finding of a positive result for P. falciparum in Alouatta from both types of forest support the hypothesis that monkeys belonging to this genus could be a potential reservoir. Furthermore, these findings raise the question of the relationship between simian and autochthonous human malaria in extra-Amazonian regions.

Malaria epidemiology in low-endemicity areas of the Atlantic Forest in the Vale do Ribeira, São Paulo, Brazil

We describe a seroepidemiological survey of malaria prevalence in two áreas of low endemicity: Intervales State Park and Alto Ribeira State Tourist Park (PETAR). Both are located in the Vale do Ribeira in the state of São Paulo, Brazil. In this study, 318 subjects from both areas had their blood analyzed for the presence of malaria parasites by thin and thick blood smears. One hundred and sixty-three (51.2%) of the subjects were from Intervales State Park and 155 (48.7%) were from PETAR. We analyzed all the samples by indirect immunofluorescent assay (IFA) to detect antibodies against asexual forms of Plasmodium vivax and Plasmodium malariae and enzyme immunosorbent assay (ELISA) to detect the presence of antibodies against circumsporozoite proteins (CSP) from P. vivax VK210, human P. vivax-like/Plasmodium simiovale, P. vivax VK247 and Plasmodium brasilianum/P. malariae. The presence of Plasmodium species was detected by polymerase chain reaction (PCR). Eighteen of the subjects analyzed had positive IFA results for IgM against P. malariae antigens, and three others were positive for P. vivax antigens. Positivity of IgG antibodies against P. vivax detected by IFA was high in samples from both Intervales State Park and PETAR (32.0% and 49.0%, respectively), while positivity for P. malariae was lower (16.0% and 19.3% in Intervales State Park and PETAR, respectively). ELISA tests showed a higher prevalence of antibodies against P. vivax VK210 (35.0%) in samples from Intervales State Park and against human P. vivax-like (29.7%) in samples from PETAR. PCR reactions revealed the presence of parasites in several of the samples analyzed. In Intervales State Park, one subject was infected by P. malariae and two by Plasmodium falciparum, while in PETAR, one subject was positive for P. falciparum and three for both P. falciparum and P. vivax parasites. The areas where these parks are located belong to the Atlantic Forest habitat, and inhabitants frequently, see monkeys. Our data suggest that monkeys may constitute a natural reservoir for malaria in both areas.

Aspectos epidemiológicos da malária humana e simiana em área de influência do reservatório da usina hidrelétrica de Porto Primavera, Município de Presidente Epitácio, Säo Paulo, Brasil / Epidemiological Aspects of Human and Simian Malaria in the Influence Area of the Reservoir of the Hydroeletric Plant at Porto Primavera, Municipality of Presidente Epitácio, State of Säo Paulo, Brazil

Objetivo: Verificar a prevalência de infecçöes maláricas, anticorpos contra formas infectantes (esporozoítos) e anticorpos contra formas sangüineas (assexuadas) de plasmódios, nas populaçöes humanas e de bugios (Alouatta caraya), que habitam a regiäo do reservatório da UHE de Porto Primavera, no Município de Presidente Epitácio, no Estado de Säo Paulo; área com história de malária autóctone. Comparar os resultados encontrados em ambas populaçöes e realizar análise epidemiológica sobre o risco da re-introduçäo da malária na regiäo e a possibilidade do envolvimento de bugios no ciclo de transmissäo. Métodos: Amostras de sangue de 216 moradores de duas áreas que se situam às margens do reservatório, Campinal (área nativa) e Reassentamento Lagoa Säo Paulo (RLSP); e amostras de sangue de 239 Bugios capturados nas matas ciliares das margens correspondentes aos lados de Säo Paulo e Mato Grosso do Sul, por ocasiäo das inundaçöes do lago artificial. Resultados: No exame parasitológico, foram encontradas formas semelhantes ao P. vivax em lâminas de gota espessa (2,9 por cento) de bugios, sendo 2 espécimes de Säo Paulo (SP) e de 5 do Mato Grosso do Sul (MS). Na técnica de PCR, foram observadas amplificaçöes em 7 amostras de DNA de bugios (4 de SP e 3 do MS) com os ôprimersö 18S r RNA gênero específico (2,9 por cento), sendo que desta, 4 amplificaram também com os ôprimersö correspondentes ao gene CS de P. vivax, e 2 com espécie específicos de P. falciparum (18S r RNA). Conclusöes: Näo foi observada a circulaçäo de plasmódios na populaçäo humana, entretanto foi verificada a presença de P. vivax e P. falciparum nos bugios. A prevalência de anticorpos contra esporozoítos (proteína CS) de plasmódios (P. vivax, P. falciparum e P. malariae), indicou que ambas populaçöes, humanas e de bugios da área de estudo, entram em contatos com estes parasitos. A prevalência de anticorpos contra formas sangüíneas indicou o desenvolvimento de infecçöes maláricas em ambas populaçöes, humanas e de bugios, sugerindo a ocorrência de quadro assintomático entre humanos e confirmando a ocorrência de infecçäo entre os macacos. Em funçäo dos resultados das pesquisas entomológicas conduzidas paralelamente na mesma área de estudo, indicarem o aumento da densidade de espécies vetoras, este quadro pode mudar, e com isso prevemos o aumento do risco da re-introduçäo da malária na populaçäo humana.