Seroprevalence of the Hepatitis E Virus in Indigenous and Non-Indigenous Communities from the Brazilian Amazon Basin.

Hepatitis E virus (HEV) infection is a common cause of acute viral hepatitis in tropical regions. In Brazil, HEV G3 is the only genotype detected to date. Reports on HEV prevalence are heterogeneous. We aimed to compare the prevalence of anti-HEV among three populations living in the Brazilian Amazon basin. Two cross-sectional studies were conducted in urban, rural, and Yanomami indigenous areas. Plasma samples from 428 indigenous and 383 non-indigenous subjects were tested for anti-HEV IgG using enzyme-linked immunosorbent assays. The overall prevalence of anti-HEV was 6.8% (95%CI: 5.25-8.72), with 2.8% (12/428) found in the Yanomami areas, 3% (3/101) in an urban area, and 14.2% (40/282) in a rural area. Multivariate logistic analysis indicated that patients aged 31-45 years or ≥46 years are more likely to present anti-HEV positivity, with a respective aOR of 2.76 (95%CI: 1.09-7.5) and 4.27 (95%CI: 1.58-12.35). Furthermore, residence in a rural area (aOR: 7.67; 95%CI: 2.50-33.67) represents a relevant risk factor for HEV infection. Additional studies detecting HEV RNA in fecal samples from both humans and potential animal reservoirs are necessary to comprehensively identify risk factors associated with HEV exposure.

Genotoxicity biomarkers on native fish species from the Araguari River (Amazon biome).

The Araguari River, one of the most important waterways in the Brazilian state of Amapá, is ecologically relevant and essential for the conservation of Amazonian fish biodiversity. Our previous studies demonstrated contamination of water and fish by metals. In particular, water samples showed genotoxic damage in Danio rerio. Here, we extended our studies of potential genotoxic damage to native fish from sampling sites located in the Araguari River lower section. To accomplish this, we collected samples of fish with different feeding behaviors, but in the same sampling sites, and evaluated the same genotoxicity biomarkers in erythrocytes. All eleven species of fish collected in the Araguari River lower section showed profiles and frequencies of genotoxic damage similar to those previously observed in tests with D. rerio, confirming that genotoxic pollutants present in these waters are also causing damage to native fish species.

Embryonic development of quail eggs (Coturnix coturnix japonica) in a homemade incubator.

Knowledge of the embryonic development of species such as quail is important for our understanding of its production and reproduction. Quail provides a practical solution to the animal protein shortage problem in developing countries and is an excellent alternative to chicken (Shanaway, 1994). This study evaluated quail embryonic viability by comparing the main morphological changes that occur over the development in a homemade incubator with previous data; the incubator was built using easily accessible and low-cost materials, which allows small producers and communities to develop quail farms as an activity to increase income or even produce a source of animal protein for the community in the Brazilian Amazon region. The homemade incubator measured 40 cm × 42 cm × 32 cm and was constructed with wooden boards, a digital thermostat, and an incandescent lamp. A total of 24 fertile eggs were incubated at a temperature of 37.5 °C and 60% relative humidity; age "0" was set at the start of incubation. Two eggs were opened on each day of development, starting from day 5 to day 16 of incubation. After gently opening the eggs, the embryos were removed, separated from the placenta and amniotic fluid, washed with running water, and weighed on a digital scale (SHIMADZU AUY 220 brand and UNI BLOC model). Crown-rump length and other measurements were performed with a digital caliper and the morphological structures were observed using a stereomicroscope (NOVA brand and NOVA ZTX-E model). All eggs opened contained live embryos, indicating that the homemade incubator temperature, relative humidity, and manual turning were adequate to maintain the quail's viability. The morphometric data of the embryos at different developmental time points were similar to those described in the literature. Embryos weighed 0.0069 g at day 5 and 4.7863 g at day 16, and the crown-rump length (CRL) means were 0.368 cm and 3.657 cm, respectively. Weight and CRL increased by 0.434 g and 2.593 cm per day of incubation, respectively. In conclusion, after 16 days of incubation in a homemade incubator, the embryos presented all phases of the development cycle, with appropriate development of weight and height based on the days of incubation without anomaly or external interference. These findings demonstrate that the homemade incubator does not alter the embryonic development of quail embryos in the Brazilian Amazon region.

O conhecimento do desenvolvimento embrionário de espécies como a codorna é importante para nosso entendimento de sua produção e reprodução. A codorna fornece uma solução prática para o problema de escassez de proteína animal nos países em desenvolvimento e é uma excelente alternativa ao frango (Shanaway, 1994). Este estudo avaliou a viabilidade embrionária de codornas comparando as principais alterações morfológicas que ocorrem ao longo do desenvolvimento em uma incubadora caseira com dados anteriores; a incubadora foi construída com materiais de fácil acesso e baixo custo, o que permite que pequenos produtores e comunidades desenvolvam criações de codornas como atividade para aumentar a renda ou mesmo produzir uma fonte de proteína animal para comunidades da Amazônia brasileira. A incubadora caseira media 40 cm × 42 cm × 32 cm e foi construída com tábuas de madeira, termostato digital e lâmpada incandescente. Um total de 24 ovos férteis foram incubados a uma temperatura de 37,5 °C e 60% de umidade relativa; idade "0" foi definida no início da incubação. Dois ovos foram abertos a cada dia de desenvolvimento, começando do dia 5 ao dia 16 de incubação. Após a abertura cuidadosa dos ovos, os embriões foram retirados, separados da placenta e do líquido amniótico, lavados em água corrente e pesados em balança digital (marca SHIMADZU AUY 220 e modelo UNI BLOC). O comprimento cabeça-cauda e demais medidas foram realizadas com paquímetro digital e as estruturas morfológicas foram observadas com auxílio de estereomicroscópio (marca NOVA e modelo NOVA ZTX-E). Todos os ovos abertos continham embriões vivos, indicando que a temperatura da incubadora caseira, a umidade relativa e a viragem manual foram adequadas para manter a viabilidade das codornas. Os dados morfométricos dos embriões em diferentes momentos do desenvolvimento foram semelhantes aos descritos na literatura. Os embriões pesaram 0,0069 g no dia 5 e 4,7863 g no dia 16, e as médias do comprimento cabeça-cauda (CRL) foram de 0,368 cm e 3,657 cm, respectivamente. Peso e CRL aumentaram 0,434 g e 2,593 cm por dia de incubação, respectivamente. Em conclusão, após 16 dias de incubação em incubadora caseira, os embriões apresentaram todas as fases do ciclo de desenvolvimento, com desenvolvimento adequado de peso e altura com base nos dias de incubação sem anomalia ou interferência externa. Esses achados demonstram que a incubadora caseira não altera o desenvolvimento embrionário de embriões de codorna na Amazônia brasileira.

Metal bioaccumulation in fish from the Araguari River (Amazon biome) and human health risks from fish consumption.

Recently, high concentrations of metals have been found in the waters from sampling sites located in the Araguari River lower section. These metals can be bioaccumulated by fish, but also biomagnified, representing a potential risk through human consumption. Here, we aimed to (1) determine the metal concentrations in muscle samples from fish; (2) evaluate bioaccumulation of metals in muscle tissue of fish species with different feeding behaviors; and (3) investigate the risks of single metals, as well as mixture of metals, to human fish consumption. Eleven fish species were sampled (3 carnivorous, 5 omnivorous, and 3 detritivorous). In four fish species, Cd and Pb concentrations exceeded the Brazilian maximum limits (MLs) for human consumption. Individually, Pb and Cd concentrations in the muscle of these fish indicate risks to human health. When considering the mixture of metals, the consumption of all fish species represents risks. The highest risks implicate detritivorous species. For all fish species evaluated, the estimated daily intake (EDI) of Hg shows values above its reference dose (RfD). Our results suggest the need to establish an environmental monitoring program that aims to preserve environmental quality, biodiversity, and human health. It is also necessary to develop actions that aim to educate Amazonian populations on safe fish preparation and consumption.

Genomic Analysis of Klebsiella pneumoniae ST258 Strain Coproducing KPC-2 and CTX-M-14 Isolated from Poultry in the Brazilian Amazon Region.

This study aimed to characterize a Klebsiella pneumoniae strain (KP411) recovered from the stool samples of poultry (Gallus gallus) in the Brazilian Amazon Region. The whole-genome sequencing of KP411 revealed the presence of an important arsenal of antimicrobial resistance genes to ß-lactams (blaCTX-M-14, blaTEM-1B, blaKPC-2, blaSVH-11), aminoglycosides [aph(3″)- Ib, aph(6)-Id, aph(3')-Ia], sulfonamides (sul1, sul2), quinolones (oqxAB), fosfomycin (fosAKP), and macrolides [mph(A)]. Furthermore, our analyses revealed that the KP411 strain belongs to the ST258 clonal lineage, which is one of the main epidemic clones responsible for the dissemination of KPC-2 worldwide. Our data suggest that food-producing animals may act as reservoirs of multidrug-resistant K. pneumoniae belonging to the ST258 clone, and, consequently, contribute to their dissemination to humans and the environment.

Behavioral patterns, parity rate and natural infection analysis in anopheline species involved in the transmission of malaria in the northeastern Brazilian Amazon region.

The characterization of behavioral patterns allows a better understanding of the transmission dynamics and the design of more effective malaria vector control strategies. This study analyzed the behavioral patterns of the Anopheles species of the Coração district situated in the northeast of the Brazilian Amazon region. The behavioral patterns of the anopheline species were measured based on the 36 collection sites of this district from December 2010 to November 2011. Collections of four hours for three consecutive nights each month and four 12-h collections, comprising two in the rainy season and two in the dry season, were performed. Furthermore, to infer the anthropophily and zoophily indexes, four additional four-hour collections were performed. The samples were also evaluated for parity rate and natural infectivity for Plasmodium spp. A total of 1689 anophelines were captured, comprising of nine species and two subgenera (Nyssorhynchus - six species, and Anopheles - three species). Anopheles darlingi was the most abundant and widely distributed species in the area, followed by A. braziliensis and A.marajoara. Anopheles darlingi and A. marajoara were the only species present in the four collections of 12-h, but only A. darlingi showed activity throughout night. Anopheles darlingi was the most anthropophilic species (AI=0.40), but the zoophily index was higher (ZI=0.60), revealing an eclectic and opportunistic behavior. Of the six most frequent species, A. nuneztovari s.l. was the most zoophilic species (ZI=1.00). All captured species showed predominance towards biting in outdoor environments. Anopheles darlingi and A. braziliensis showed multimodal biting peaks, whereas A. marajoara revealed a stable pattern, with the biting peak after sunset. Using the PCR technique, no anopheline was found infected with the malaria parasite. Since A. darlingi and A. marajoara are recognized as important vectors in this region, the district of Coração may be considered as a highly potent area for transmission of malaria, therefore, the prevention and surveillance measures should be taken constantly to prevent the same. The role of A. braziliensis as malaria vector needs to be urgently investigated.

JC virus/human immunodeficiency virus 1 co-infection in the Brazilian Amazonian region

Abstract JC virus (JCV) is a member of the Polyomaviridae family and is associated to a severe disease known as progressive multifocal leukoencephalopathy, PML, which is gradually increasing in incidence as an opportunistic infection among AIDS patients. The present study aimed to investigate the occurrence of JCV among HIV-1 carriers including their types and molecular subtypes and the possible association with disease. Urine samples from 66 HIV-1 infected subjects were investigated for the presence of the virus by amplifying VP1 (215 bp) and IG (610 bp) regions using the polymerase chain reaction. JCV was detected in 32% of the samples. The results confirmed the occurrence of type B (subtype Af2); in addition, another polyomavirus, BKV, was also detected in 1.5% of samples of the HIV-1 infected subjects. Apparently, there was no significant difference between mono- (HIV-1 only) and co-infected (HIV-1/JCV) subjects regarding their TCD4+/TCD8+ lymphocyte counts or HIV-1 plasma viral load. Self admitted seizures, hearing and visual loses were not significantly different between the two groups.

JC virus/human immunodeficiency virus 1 co-infection in the Brazilian Amazonian region.

JC virus (JCV) is a member of the Polyomaviridae family and is associated to a severe disease known as progressive multifocal leukoencephalopathy, PML, which is gradually increasing in incidence as an opportunistic infection among AIDS patients. The present study aimed to investigate the occurrence of JCV among HIV-1 carriers including their types and molecular subtypes and the possible association with disease. Urine samples from 66 HIV-1 infected subjects were investigated for the presence of the virus by amplifying VP1 (215bp) and IG (610bp) regions using the polymerase chain reaction. JCV was detected in 32% of the samples. The results confirmed the occurrence of type B (subtype Af2); in addition, another polyomavirus, BKV, was also detected in 1.5% of samples of the HIV-1 infected subjects. Apparently, there was no significant difference between mono- (HIV-1 only) and co-infected (HIV-1/JCV) subjects regarding their TCD4(+)/TCD8(+) lymphocyte counts or HIV-1 plasma viral load. Self admitted seizures, hearing and visual loses were not significantly different between the two groups.

Surveillance, health promotion and control of Chagas disease in the Amazon Region - Medical attention in the Brazilian Amazon Region: a proposal

We refer to Oswaldo Cruz’s reports dating from 1913 about the necessities of a healthcare system for the Brazilian Amazon Region and about the journey of Carlos Chagas to 27 locations in this region and the measures that would need to be adopted. We discuss the risks of endemicity of Chagas disease in the Amazon Region. We recommend that epidemiological surveillance of Chagas disease in the Brazilian Amazon Region and Pan-Amazon region should be implemented through continuous monitoring of the human population that lives in the area, their housing, the environment and the presence of triatomines. The monitoring should be performed with periodic seroepidemiological surveys, semi-annual visits to homes by health agents and the training of malaria microscopists and healthcare technicians to identify Trypanosoma cruzifrom patients’ samples and T. cruziinfection rates among the triatomines caught. We recommend health promotion and control of Chagas disease through public health policies, especially through sanitary education regarding the risk factors for Chagas disease. Finally, we propose a healthcare system through base hospitals, intermediate-level units in the areas of the Brazilian Amazon Region and air transportation, considering the distances to be covered for medical care.

Hepatite crônica B oculta: prevalência e aspectos clínicos em população de elevada endemicidade de infecção pelo vírus da hepatite B na Amazônia Ocidental Brasileira / Occult hepatitis B: prevalence and clinical characteristics in a population with high endemicity of hepatitis B infection in the western Brazilian Amazon region

Recentemente é descrito estado de persistência do vírus da hepatite B denominado hepatite crônica B oculta. Sua prevalência e fisiopatologia são desconhecidas. O objetivo deste estudo foi avaliar a ocorrência dessa entidade clínica em pacientes da Amazônia brasileira. De 51 pacientes anti-HBc total reativos testados pela reação em cadeia da polimerase, 17 por cento foram positivos. Não observamos associação com fatores de risco clássicos de infecção pelo vírus da hepatite B, testes bioquímicos, hematológicos e histopatologia. No entanto, os pacientes ictéricos e reativos para o anti-HIV apresentaram associação com a presença do ADN-vírus da hepatite B. Os resultados demonstram a ocorrência da hepatite crônica B oculta, entre nossos doentes, porém, com taxas de prevalência abaixo do esperado para a região. Acreditamos que, apesar do tamanho da amostra avaliada ser pequeno, sua ocorrência poderia ter sido maior se empregássemos primers para a região S, C e X do genoma do vírus da hepatite B, aumentando a sensibilidade do teste.

A persistent form of the hepatitis B virus called occult chronic hepatitis B has recently been described. Its prevalence and physiopathology are unknown. The aim of this study was to evaluate the occurrence of this clinical entity among patients in the Brazilian Amazon region. Out of 51 anti-HBc total-positive patients who were tested using the polymerase chain reaction, 17 percent were positive. We did not find any associations with classical risk factors for hepatitis B virus infection or with biochemical tests, hematological tests or histological patterns. However, the jaundiced and HIV-positive patients showed a statistical association with the presence of hepatitis B virus-DNA. The results demonstrated that occult hepatitis B occurred among our patients, but at prevalence rates lower than expected for this region. We believe that despite the small sample size, the occurrence might have been found to be greater if we had used primers for the S, C and X regions of the hepatitis B virus genome, thereby increasing the sensitivity of the test.

Frequency of the HFE C282Y and H63D polymorphisms in Brazilian malaria patients and blood donors from the Amazon region

Malaria is an endemic parasitosis and its causitive agent, Plasmodium, has a metabolism linked to iron supply. HFE is a gene with the polymorphisms C282Y and H63D, which are associated with a progressive iron accumulation in the organism leading to a disease called hereditary hemochromatosis. The aim of the present study was to determine the allelic and genotypic frequencies of the HFE gene polymorphisms in malaria patients and blood donors from the Brazilian Amazon region. We screened 400 blood donors and 400 malaria patients for the HFE C282Y and H63D polymorphisms from four states of the Brazilian Amazon region by polymerase chain reaction and restriction fragment length polymorphism analysis. We did not find any C282Y homozygous individuals, and the only five heterozygous individuals detected were from Pará State. The most frequent genotype in the North region of Brazil was the H63D heterozygote, in both study groups. Our results contribute to the concept that the Brazilian Amazon region should not be regarded as a single entity in South America. These polymorphisms did not influence the symptoms of malaria in the population studied, as neither severe signs nor high parasitemia were observed. Therefore, different hereditary hemochromatosis diagnostic and control measures must be developed and applied within its diverse locations. Investigations are currently being carried out in our laboratory in order to determine the importance of the coexistence of hereditary hemochromatosis in patients affected by parasitic diseases, such as malaria.