Detection of etiological agents of malaria in howler monkeys from Atlantic Forests, rescued in regions of São Paulo city, Brazil.

BACKGROUND: In some states of the Brazilian extra-Amazonian region, such as the Atlantic Forest area, autochthonous human cases of malaria were related to simian malarias and vice versa. METHODS: To verify the presence of Plasmodium, 50 blood samples of howler monkeys (Alouatta guariba clamitans) rescued from the Metropolitan Region of Saõ Paulo city, where the Atlantic Forest is present, were analyzed. The samples were submitted to microscopy (thin and thick blood smears), enzyme-linked immunosorbent assays (ELISA), indirect immunofluorescent assay (IFA), and polymerase chain reaction (PCR). RESULTS: Only one smear showed forms reminiscent of Plasmodium vivax. In ELISA, the frequencies of antibodies against synthetic peptides corresponding to circumsporozoite protein of P. vivax VK210 'classic' (Pvc), P. vivax VK247, human P. vivax-like (Pvk and Pvl), P. malariae/P. brasilianum (Pm), and P. falciparum (Pf) were 24.0% (12/50) for Pvc, 8.0% (04/50) for Pvk, 6.0% (03/50) for Pvl, 24.0% (12/50) for Pm, and 28.0% (14/50) for Pf, while the frequency of antibodies against PvMSP119 recombinant proteins was 42.0% (21/50). No serum reacted against PfMSP1-19. In IFA,the seropositivity of antibodies against asexual forms of P. malariae was 31.3% (15/48). We utilized three PCR protocols to develop a molecular consensus (positive results in, at least, two protocols). The frequency of Plasmodium infections detected by PCR was 18.0% (09/50) for P. vivax, 4.0% (02/50) for P. malariae, and 76.0% (38/50) of samples were negative. The molecular consensus was not seen in 4.0% (02/50) of samples. CONCLUSIONS: These results suggest that a possible interaction between human and simian malaria coming from a zoonotic cycle cannot be discarded because simians that live in the areas of the Atlantic Forest could play a role as a reservoir for Plasmodium.

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.

Entomological characterization and natural infection of anophelines in an area of the Atlantic Forest with autochthonous malaria cases in mountainous region of Espírito Santo State, Brazil.

Autochthonous malaria cases in the state of Espírito Santo, Brazil, are distributed in mountainous regions surrounded by the Atlantic Forest. While some aspects of this disease are unclear, detection of possible vector species can help to elucidate epidemiological uncertainties. Entomological and natural infection studies were carried out using anophelines (Diptera: Culicidae) captured in the municipality of Santa Tereza, ES. Monthly captures were made from March 2004 to February 2006. CDC-CO2 traps were used from dusk (6:00 P.M.) to dawn (6:00 A.M.) to capture anophelines in the following habitats: near the houses, in open areas (at ground level) and inside, and at the margins of the forest (canopy and ground level). Shannon light traps were also used at the same locations of the CDC-CO2 traps. A total of 2,290 anophelines within 10 species were captured. The relative frequency of Anopheles (Kerteszia) cruzii Dyar & Knab / A.(K.) homunculus Komp was the highest, with the majority captured in CDC-CO2 traps installed in the forest canopy. The main species captured in Shannon traps was A.(Nyssorhynchus) strodei Root. The largest number of anophelines was captured from July to September and from 6:00 P.M. to 10:00 P.M. Anopheles (K.) cruzii is the probable vector for malaria transmission inside or near the Atlantic Forest fragments, but the role of other species cannot be ignored, as 53% of the sampled anophelines belonged to the subgenus Nyssorhynchus. The natural infection of A. cruzii, A. parvus (Chagas) and A. galvaoi Causey, Deane & Deane by Plasmodium vivax detected by PCR from DNA extracted from their thoraxes supports this view.

Entomological characterization and natural infection of anophelines in an area of the Atlantic Forest with autochthonous malaria cases in mountainous region of Espírito Santo State, Brazil / Caracterização entomológica e infecção natural de anofelinos em área de Mata Atlântica, com casos autóctones de malária, em regiões montanhosas do Espírito Santo

Autochthonous malaria cases in the state of Espírito Santo, Brazil, are distributed in mountainous regions surrounded by the Atlantic Forest. While some aspects of this disease are unclear, detection of possible vector species can help to elucidate epidemiological uncertainties. Entomological and natural infection studies were carried out using anophelines (Diptera: Culicidae) captured in the municipality of Santa Tereza, ES. Monthly captures were made from March 2004 to February 2006. CDC-CO2 traps were used from dusk (6:00 P.M.) to dawn (6:00 A.M.) to capture anophelines in the following habitats: near the houses, in open areas (at ground level) and inside, and at the margins of the forest (canopy and ground level). Shannon light traps were also used at the same locations of the CDC-CO2 traps. A total of 2,290 anophelines within 10 species were captured. The relative frequency of Anopheles (Kerteszia) cruzii Dyar & Knab / A. (K.) homunculus Komp was the highest, with the majority captured in CDC-CO2 traps installed in the forest canopy. The main species captured in Shannon traps was A. (Nyssorhynchus) strodei Root. The largest number of anophelines was captured from July to September and from 6:00 P.M. to 10:00 P.M. Anopheles (K.) cruzii is the probable vector for malaria transmission inside or near the Atlantic Forest fragments, but the role of other species cannot be ignored, as 53 percent of the sampled anophelines belonged to the subgenus Nyssorhynchus. The natural infection of A. cruzii, A. parvus (Chagas) and A. galvaoi Causey, Deane & Deane by Plasmodium vivax detected by PCR from DNA extracted from their thoraxes supports this view.

No Espírito Santo, os casos de malária autóctone estão distribuídos na região serrana próximo aos fragmentos de Mata Atlântica. Uma vez que alguns aspectos da doença são obscuros, a detecção das possíveis espécies de vetores pode auxiliar na elucidação de incertezas epidemiológicas. Estudos entomológicos e de infecção natural foram realizados com anofelinos (Diptera: Culicidae) capturados no município de Santa Tereza, ES. Capturas mensais foram realizadas de março de 2004 a fevereiro de 2006. Armadilhas CDC-CO2 foram utilizadas do crepúsculo (18:00h) ao amanhecer (6:00h), para capturar anofelinos nos seguintes habitats: próximo ao domicílio e área aberta (solo), margem e interior da mata (solo e copa). Armadilhas Shannon também foram utilizadas nos mesmos locais que as de CDC-CO2. Capturou-se o total de 2.290 anofelinos distribuídos em 10 espécies. A maior frequência relativa foi de Anopheles (Kerteszia) cruzii Dyar & Knab / A. (K.) homunculus Komp, sendo a maioria capturada em CDC-CO2 instalada na copa da mata. A principal espécie capturada em armadilha Shannon foi A.(Nyssorhynchus) strodei Root. O maior número de anofelinos foi capturado entre julho e setembro das 18:00h às 22:00h. Provavelmente A. (K.) cruzii é responsável pela transmissão da malária dentro ou próximo aos fragmentos de Mata Atlântica. Entretanto, a participação de outras espécies não pode ser ignorada, visto que 53 por cento da amostragem foi constituída pelo subgênero Nyssorhynchus. A detecção de Plasmodium vivax no tórax de A. cruzii, A. parvus (Chagas) e A. galvaoi Causey, Deane & Deane por meio de PCR reforça esse argumento.

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 por cento or 8/71 (5.6 por cento for Plasmodium malariae and 5.6 por cento for Plasmodium vivax) and one specimen was positive for Plasmodium falciparum (1.4 por cento); 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 por cento (16/235) in the PCR tests (5.5, 0.8 and 0.4 por cento 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 por cento). 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...(AU)