Enteric viruses' dissemination in a private reserve of natural heritage.

This study aimed to assess anthropogenic impact of surrounding population in the Private Reserve of Natural Heritage at Pantanal, the world's largest freshwater wetland ecosystem located in the centre of South America. Viral aetiological agents of acute gastroenteritis as rotavirus A (RVA), noroviruses, human adenoviruses, klassevirus and of hepatitis, as hepatitis A virus, were investigated in different aquatic matrices. Annual collection campaigns were carried out from 2009 to 2012, alternating dry and rainy seasons. Viral particles present in the samples were concentrated by the adsorption-elution method, with negatively charged membranes, and detected by qualitative and quantitative PCR. From a total of 43 samples at least one virus was detected in 65% (28) of them. Viruses were detected in all matrices with concentrations ranging from 2 × 102 to 8·3 × 104 genome copies per litre. A significant higher RVA frequency was observed in the dry season. Our data revealing dissemination of human enteric viruses in water matrices both inside and outside the reserve could be useful to trace faecal contamination in the environment and to minimize the risk of infection by exposure of susceptible individuals. SIGNIFICANCE AND IMPACT OF THE STUDY: This study is part of a collaborative project designed to investigate the environmental and health conditions of the Private Reserve of Natural Heritage at Pantanal, the largest seasonally flooded wetland in the world. The project aimed to promote health and quality of human and wildlife extending technical-scientific knowledge about pathogens present in the region. By assessing the occurrence of human enteric viruses in different water matrices we demonstrated the anthropogenic impact of surrounding population and pointed out the potential risk of infection by exposure of susceptible individuals.

Estudo anatômico do plexo lombossacral de Tamandua tetradactyla / Anatomical study of the lumbosacral plexus of the Tamandua tetradactyla

O tamanduá-mirim (Tamandua tetradactyla) é um xenartro da família Myrmecophagidae, encontrado da Venezuela ao sul do Brasil. Estudos apontam que essa é uma das espécies de animais selvagens mais vitimadas em número de atropelamentos, e, muitas vezes, o atendimento clínico adequado aos indivíduos feridos é dificultado pela carência de informações acerca dos mesmos. Visando contribuir com o conhecimento dessa espécie, este estudo teve como objetivo descrever seu plexo lombossacral. Para tanto, foram utilizados quatro cadáveres de Tamandua tetradactyla adultos e de ambos os sexos. O plexo lombossacral dessa espécie é formado pelos ramos ventrais dos nervos espinhais T18, L1, L2, L3, S1, S2, S3, S4, S5. Os nervos integrantes do plexo lombossacral do T. tetradactyla com suas formações mais frequentes foram os seguintes: genitofemoral (T18), cutâneo femoral lateral (T18-L1), femoral (T18, L1-L3), obturador (T18, L1-L3), glúteo cranial (L3-S1), isquiático (L3-S3), pudendo (S3-S4 ou S4-S5), retal caudal (S4 ou S5) e cutâneo femoral caudal (S4-S5). O plexo lombar e sacral dessa espécie é unido, sendo L3 o ponto de união entre eles. Devido ao pequeno número de vértebras lombares, a composição dos nervos do plexo lombossacral do T. tetradactyla apresenta características peculiares que se diferem das características das demais espécies já estudadas, quais sejam, a ausência dos nervos ílio-hipogástrico e ilioinguinal e participação de nervos torácicos na composição dos nervos do plexo lombar, presença de contribuição sacral na composição do nervo obturador e ausência de contribuição lombar na composição do nervo isquiático e um limite mais caudal na extensão do plexo sacral.

The lesser anteater (Tamandua tetradactyla) is a xenarthra of the Myrmecophagidae family found from Venezuela to southern Brazil. Studies have shown that this is one of the most numerous wildlife species victims of car collisions on roads, and often the appropriate clinical care to injured animals is hindered by the lack of information about them. In order to contribute to the knowledge of this species, this study aimed to describe its lumbosacral plexus. For this purpose, four cadavers of adult specimens of both sexes of T. tetradactyla were used. The lumbosacral plexus of the T. tetradactyla is formed by the ventral rami of spinal nerves T18, L1, L2, L3, S1, S2, S3, S4, S5. The lumbosacral plexus nerves with their most common formations in this species were as follows: genitofemoral (T18), lateral femoral cutaneous (T18-L1), femoral (T18, L1-L3), obturator (T18, L1-L3), cranial gluteal (L3-S1), ischiatic (L3-S3), pudendus (S3-S4 or S4-S5), caudal retal (S4 or S5), and caudal femoral cutaneous (S4-S5). The lumbar and sacral plexus of this species is joined, L3 being the link between them. Due to the small number of lumbar vertebrae, the arrangement of the lumbosacral plexus nerves of the T. tetradactyla showed peculiar characteristics that differ it from that of other previously studied species, such as the absence of iliohypogastric and ilioinguinal nerves and contribution of thoracic nerves in the formation of all the nerves of the lumbar plexus, presence of sacral contribution in the formation of the obturator nerve, and the lack of lumbar contribution for sciatic nerve formation and a most caudal extent of the sacral plexus.

Environmental dissemination of group A rotavirus: P-type, G-type and subgroup characterization.

Rotaviruses A (RV-A) infection is the most common cause of acute diarrheal diseases in infants and the dissemination of these viruses in the environment represents a public health hazard. The present study aims to evaluate reverse transcription-polymerase chain reaction (RT-PCR) based protocols for the detection of RV-A genes in different types of environmental samples. RV-A were concentrated by the adsorption-elution method using negatively charged membranes associated with a Centriprep Concentrator 50. The RV-A VP4, VP7 and VP6 genes were detected using RT-PCR in river water from the Amazon Hydrographic basin (Northern region) and from wastewater in a sewage treatment plant in Rio de Janeiro (Southeast region), Brazil. RV-A were successfully detected in water environmental samples by the methods used. The detection of the VP6 gene by RT-PCR was the most sensitive for detecting RV-A in environmental samples (44.0%), when compared to the detection of the VP4 (33.3%) and VP7 (25.3%) genes. Based on nucleotide sequence and phylogenetic analysis of the partial VP6 gene, 22 environmental samples were determined to be subgroup II (Wa-like). These results indicate that analysis of environmental samples could possibly make a valuable contribution to studies on the epidemiology of RV-A.

Molecular detection of hepatitis A virus in urban sewage in Rio de Janeiro, Brazil.

AIMS: A one-year survey was conducted to examine hepatitis A virus (HAV) prevalence, distribution of genotypes and their relationship to bacterial indicators in raw and treated sewage samples. METHODS AND RESULTS: Fifty sewage samples (raw = 25 and treated = 25) were collected twice monthly from one sewage treatment plant in Rio de Janeiro. Virus concentration was performed by adsorption to an electronegative membrane followed by ultrafiltration. Viral RNA was detected by nested reverse transcriptase-polymerase chain reaction (RT-PCR) and real-time PCR and positive products were directly sequenced. Total and faecal coliform concentrations were also determined. By nested RT-PCR, HAV RNA was detected in 16/50 (32%) and eight (16%) of them were found in treated sewage samples. By real-time PCR, HAV RNA was detected in 46/50 (92%) samples and 24 were from treated sewage. Phylogenetic analyses classified nine isolates (56%) as subgenotype IA and seven (44%) as IB. CONCLUSIONS: Real-time PCR was more sensitive than nested RT-PCR; the presence of subgenotypes IA and IB was described and bacterial indicators cannot be used to predict HAV presence in sewage. SIGNIFICANCE AND IMPACT OF THE STUDY: These results demonstrated that HAV still remains in the environment after sewage treatment and could play an important role in maintaining the endemicity of HAV infection.

Dengue virus surveillance: the co-circulation of DENV-1, DENV-2 and DENV-3 in the State of Rio de Janeiro, Brazil.

This study presents the results obtained in the monitoring of dengue virus (DENV) transmission in the Greater Metropolitan Region of the State of Rio de Janeiro, in the period 2000-2001. A total of 5324 serum samples from suspected cases of dengue were analysed in order to confirm dengue infection. The introduction of DENV-3 to the region in December 2000 resulted in the co-circulation of three serotypes: DENV-1, DENV-2 and DENV-3. In this study, virus isolation and/or reverse transcriptase PCR (RT-PCR) confirmed 52.3% (42/79) of DENV-3 cases, showing the importance of acute serum samples in the virological surveillance of the disease. Despite the introduction of a new serotype, an outbreak due to DENV-1 was observed in the municipality of Niteroi. The restriction site-specific PCR (RSS-PCR) patterns obtained for DENV-1 and DENV-2 isolated in that period showed that those strains belonged to the subtypes previously circulating in the state. DENV-3 RSS-PCR patterns confirmed that these viruses belonged to subtype C (Sri Lanka/India strains), represented by the strain circulating on the American continent. These data showed the importance of an active surveillance programme in countries where dengue is endemic.