Abstract: INTRODUCTION : Molecular analyses are auxiliary tools for detecting Koch's bacilli in clinical specimens from patients with suspected tuberculosis (TB). However, there are still no efficient diagnostic tests that combine high sensitivity and specificity and yield rapid results in the detection of TB. This study evaluated single-tube nested polymerase chain reaction (STNPCR) as a molecular diagnostic test with low risk of cross contamination for detecting Mycobacterium tuberculosis in clinical samples. METHODS: Mycobacterium tuberculosis deoxyribonucleic acid (DNA) was detected in blood and urine samples by STNPCR followed by agarose gel electrophoresis. In this system, reaction tubes were not opened between the two stages of PCR (simple and nested). RESULTS: STNPCR demonstrated good accuracy in clinical samples with no cross contamination between microtubes. Sensitivity in blood and urine, analyzed in parallel, was 35%-62% for pulmonary and 41%-72% for extrapulmonary TB. The specificity of STNPCR was 100% in most analyses, depending on the type of clinical sample (blood or urine) and clinical form of disease (pulmonary or extrapulmonary). CONCLUSIONS: STNPCR was effective in detecting TB, especially the extrapulmonary form for which sensitivity was higher, and had the advantage of less invasive sample collection from patients for whom a spontaneous sputum sample was unavailable. With low risk of cross contamination, the STNPCR can be used as an adjunct to conventional methods for diagnosing TB.

Blood infection by the simian parasite, Plasmodium simium, was identified in captive (n = 45, 4.4%) and in wild Alouatta clamitans monkeys (n = 20, 35%) from the Atlantic Forest of southern Brazil. A single malaria infection was symptomatic and the monkey presented clinical and haematological alterations. A high frequency of Plasmodium vivax-specific antibodies was detected among these monkeys, with 87% of the monkeys testing positive against P. vivax antigens. These findings highlight the possibility of malaria as a zoonosis in the remaining Atlantic Forest and its impact on the epidemiology of the disease.

Malária continua sendo uma das mais importantes doenças endêmicas da região Amazônica. Esse estudo mostra o impacto da colonização humana sobre a estrutura populacional de Anopheles. Diversidade, abundância, riqueza e distribuição do gênero Anopheles foi observado em duas áreas com diferentes graus de antropização no município do Cantá, Estado de Roraima, Norte do Brasil. A influência das estações seca e chuvosa sobre a população dos mosquitos também foram investigadas. A captura dos mosquitos foi realizada entre 6:00 e 10:00 pm na estação seca (Fevereiro e Novembro) e estação chuvosa (Maio e Agosto) em quatro locais diferentes. Entre as 11 espécies de Anopheles identificadas através das características dos adultos, An. albitarsis s.l. (45.5%) e An. darlingi (19.2%) foram as mais abundantes nas áreas mais intensamente antropizadas, enquanto An. triannulatus (19.2%) foi a espécie mais comum na área menos modificada. As outras espécies encontradas foram An. nuneztovari (10.9%), An. oswaldoi (2.0%), An. evansae (1.7%), An. brasiliensis (0.6%), An. intermedius (0.3%), An. mediopunctatus (0.5%), An. periassui (0.08%) e An. argyritarsis (0.04%). A mais alta densidade populacional de mosquitos foi observada em Maio e as mais baixas em Fevereiro e Novembro. Esses resultados demonstram a existência de uma alta diversidade de anofelinos na área estudada e que mudanças antrópicas no ambiente e variabilidade climática afetam a densidade populacional desses vetores.

Malaria has still been one of the most important endemic diseases in the Amazonian region. This study presents the impact of human settlements on the structure of Anopheles population. Diversity, abundance, richness and distribution of the genus Anopheles were observed in two areas with different levels of human settlement in the Cantá city, Roraima State, Northern Brazil. The influence of the dry and rainy seasons on mosquito populations was also observed. Mosquito captures were performed between 6:00 and 10:00 pm during the dry (February and November) and rainy (May and August) seasons at four different sites of each area. Among the 11 species of Anopheles identified through the adults' characteristics, An. albitarsis s.l. (45.5%) and An. darlingi (19.2%) were the most abundant in the more intensively anthropized area while An. triannulatus (19.2%) was more common in the less modified area. Other species found were An. nuneztovari (10.9%), An. oswaldoi (2.0%), An. evansae (1.7%), An. brasiliensis (0.6%), An. intermedius (0.3%), An. mediopunctatus (0.5%), An. periassui (0.08%) and An. argyritarsis (0.04%). The highest mosquitoes' population density was observed in May and the lowest one was observed in February and November. These results demonstrate the existence of a high diversity of anophelines in the study areas, showing that anthropic changes in the environment and climate variability affect both the population density and relative abundance of these vectors.

Understanding the different background landscapes in which malaria transmission occurs is fundamental to understanding malaria epidemiology and to designing effective local malaria control programs. Geology, geomorphology, vegetation, climate, land use, and anopheline distribution were used as a basis for an ecological classification of the state of Roraima, Brazil, in the northern Amazon Basin, focused on the natural history of malaria and transmission. We used unsupervised maximum likelihood classification, principal components analysis, and weighted overlay with equal contribution analyses to fine-scale thematic maps that resulted in clustered regions. We used ecological niche modeling techniques to develop a fine-scale picture of malaria vector distributions in the state. Eight ecoregions were identified and malaria-related aspects are discussed based on this classification, including 5 types of dense tropical rain forest and 3 types of savannah. Ecoregions formed by dense tropical rain forest were named as montane (ecoregion I), submontane (II), plateau (III), lowland (IV), and alluvial (V). Ecoregions formed by savannah were divided into steppe (VI, campos de Roraima), savannah (VII, cerrado), and wetland (VIII, campinarana). Such ecoregional mappings are important tools in integrated malaria control programs that aim to identify specific characteristics of malaria transmission, classify transmission risk, and define priority areas and appropriate interventions. For some areas, extension of these approaches to still-finer resolutions will provide an improved picture of malaria transmission patterns.

Examinando-se o sangue de 2.046 primatas capturados durante a "Operação Curupira" encontraram-se plasmódios semelhantes ao Plasmodium brasilianum em sete espécies ou subespécies: Alouatta belzebul belzebul, Alouatta belzebul nigerrima, Alouatta seniculus, Chiropotes satanas, Callicebus moloch, Saimiri sciureus e Saguinus midas niger. Esta última espécie havia sido ainda encontrada naturalmente parasitada por plasmódios.

Samples of blood taken from 2.046 primates relocated by the "Curupira project" were examined for the presence of simian malaria parasites. This project aimed to rescuing the animals that lived within the forest that began to be flooded by the Tucurui dam built in the Tocantins river, Amazon basin, Brazil. Malaria parasites similar or identical to Plasmodium brasilianum were found in seven species or subspecies: Alouatta belzebul belzebul, A. belzebul nigerrima, A. seniculus, Chiropotes satanas, Saimiri sciureaus, Callicebus moloch and Saguinus midas niger. This last species was never reported infected with malaria parasites before.