Viral gastroenteritis and other waterborne diseases are a major concern for health in Brazil. A number of studies were conducted about the presence of viruses on water samples from Brazilian areas. However, the knowledge about the occurrence of viral contamination of drinking water sources in rural settings of the country is insufficient. On the present work, 15 samples from 5 dairy farms located at the municipality of Tenente Portela were collected and analysed for the presence of human adenoviruses (HAdV), as well as human enteroviruses (EV) and rotaviruses (RV). HAdV was present on 66.66% of the water samples, and have been found in all samples from artesian wells and springs, which are used as sources of drinking water for the individuals inhabiting those farms. EV and RV found only in one sample each. The detection rates of HAdV on the water from these dairy farms are alarming and point towards a situation of elevated environmental contamination by fecal microorganisms of human origin and poor basic sanitation conditions.
Around the world, enteric viruses are often found in surface waters. This study set out to evaluate the occurrence of adenoviruses (AdVs) in water samples, and its relation to different physical, chemical, and bacteriological parameters [total coliform (TC) and fecal coliform (FC), represented by Escherichia coli]. Monthly samples of 500 ml of raw water were collected from May 2011 to June 2013 in eight abstraction points water treatment stations along three stretches of the Sinos River Basin (SRB), in Southern Brazil and, subsequently, were analyzed using real-time polymerase chain reaction (qPCR). AdVs from different species, from human (HAdV), and from other animals (CAV1-2, BAdV, PAdV, and AvAdV) were detected along the three stretches of the basin, indicating fecal contamination from different sources and proving the inefficiency of the wastewater treatment in the waters of the SRB and intensifying the strong influence of human activities that can contribute to the presence of inhibitory substances such as organic acids in surface of these waters. Statistical analyses revealed no significant correlations between the concentrations of TC and FC and the concentrations of AdVs. We observed a small, nonconstant, and unstable correlation between viruses and physicochemical parameters. These correlations were not sufficiently consistent to establish a reliable association; therefore, this study corroborates that only the viral assay itself is reliable for the diagnosis of fecal contamination by viruses in environmental samples.
Hepatitis E virus (HEV) is highly disseminated among swine herds worldwide. HEV is also a threat to public health, since particularly genotypes 3 and 4 may cause acute hepatitis in human beings. No previous studies were done on the occurrence of HEV in environmental samples in Rio Grande do Sul, Brazil. In the present study, reverse transcriptase-polymerase chain reaction (RT-PCR) was employed to detect the presence of HEV in swine feces and in effluents from slurry lagoons in farms located in the municipality of Teutônia, inside the area of swine husbandry in the state. Pooled fecal samples from the floor of pig barns from 9 wean-to-finish farms and liquid manure samples were collected from the slurry lagoons from 8 of these farms. From the pooled fecal samples, 8/9 were positive for the HEV ORF1 gene by RT-PCR; all the slurry lagoon samples were positive for HEV RNA (100%). The identity of the HEV ORF1 amplicons was confirmed by sequencing belonging to HEV genotype 3, which was previously shown to be circulating in South America.Keywords: Hepatitis E virus, swine HEV, slurry, zoonosis RESUMOO vírus da hepatite E (HEV) é altamente disseminado entre rebanhos suínos no mundo todo. O HEV é também uma ameaça à saúde pública, já que os genótipos 3 e 4 podem causar hepatite aguda em seres humanos. Não há estudos anteriores sobre a ocorrência de HEV em amostras ambientais no Rio Grande do Sul. No presente estudo, empregou-se transcrição reversa e reação em cadeia da polimerase (RT-PCR) para detectar a presença de HEV em fezes de suínos e efluentes de lagoas de chorume em fazendas localizadas no município de Teutônia, representativo da região de maior produção de suínos no estado. Pools de amostras fecais foram coletadas a partir do chão de galpões de suínos provenientes de 9 propriedades de terminação; outra amostra de esterco líquido foi coletada das lagoas de chorume de 8 dessas fazendas. A partir das amostras fecais reunidas, 8/9 foram positivas para o gene ORF1 de HEV por PCR convencional; todas as amostras de lagoas de chorume foram positivas para RNA de HEV (100%). A identificação dos produtos de amplificação de HEV ORF1 foi confirmada por sequenciamento pertencente ao HEV genótipo 3, o qual foi previamente detectado na América do Sul.
Human astrovirus (HAstV) constitutes a major cause of acute gastroenteritis in children. The viral 5′ and 3′ untranslated regions (UTR) have been involved in the regulation of several molecular mechanisms. However, in astrovirues have been less characterized. Here, we analyzed the secondary structures of the 5′ and 3′ UTR of HAstV, as well as their putative target sites that might be recognized by cellular factors. To our knowledge, this is the first bioinformatic analysis that predicts the HAstV 5′ UTR secondary structure. The analysis showed that both the UTR sequence and secondary structure are highly conserved in all HAstVs analyzed, suggesting their regulatory role of viral activities. Notably, the UTRs of HAstVs contain putative binding sites for the serine/arginine-rich factors SRSF2, SRSF5, SRSF6, SRSF3, and the multifunctional hnRNPE2 protein. More importantly, putative binding sites for PTB were localized in single-stranded RNA sequences, while hnRNPE2 sites were localized in double-stranded sequence of the HAstV 5′ and 3′ UTR structures. These analyses suggest that the combination of SRSF proteins, hnRNPE2 and PTB described here could be involved in the maintenance of the secondary structure of the HAstVs, possibly allowing the recruitment of the replication complex that selects and recruits viral RNA replication templates.
In the present study, nine coagulants having potential to be used for sewage treatment were compared to assess their efficiency in removing total coliform bacteria, Escherichia coli and adenovirus. The coagulants tested were metallic and organic and their efficiency was compared when treating samples of raw and treated sewage (activated sludge). Before the efficiency tests of the coagulants, viral concentration methods were compared. Coagulation tests were carried out by using the jar-test system and the doses used ranged from 100 ppm to 1,000 ppm. Viral DNA was extracted and subjected to real-time polymerase chain reaction (qPCR) using primers for the gene of AdV hexon. Aluminum sulfate (1,000 ppm) presented the best results for raw sewage among metal coagulants whereas Acquapol C118 and WW (1,000 ppm) had the most satisfactory results among organic coagulants, both reducing up to 7 logs for coliforms and 4 logs for virus. For the treated effluent, FeCl (1,000 ppm) presented best results for metal coagulants, whereas, from organic coagulants, the best removal rates were for Acquapol 893/11 (1,000 ppm), both reducing up to 3 logs for coliforms and 4 logs for virus.
Adenoviruses are among the most promising viral markers of fecal contamination. They are frequently found in the water, sediment and soil of regions impacted by human activity. Studies of the bioaccumulation of enteric viruses in shrimp are scarce. The cities located in the northern coast of the lake systems in Southern Brazil have high urbanization and intensive farming rates, and poor sewage collection and treatment. One hundred (n = 100) Farfantepenaeus paulensis pink-shrimp specimens and 48 water samples were collected from coastal lagoons between June 2012 and May 2013. Water samples were concentrated and the shrimp, mashed. After DNA extraction, samples were analyzed by real time polymerase chain reaction (qPCR) in order to detect and quantify viral genomes. Thirty-five percent of shrimp samples were positive for contamination, predominantly by avian adenoviruses. A total of 91.7% of water samples contained adenoviruses DNA, with the human form being the most frequent. Our results provided evidence of significant bioaccumulation of adenoviruses in shrimp, showing the extent of the impact of fecal pollution on aquatic ecosystems.
RESUMOO abastecimento de água do município de Ivoti provém unicamente de origem subterrânea. A potabilidade da água subterrânea do município foi avaliada através da análise de marcadores bacterianos (Escherichia coli e coliformes totais) e virais (adenovírus, enterovírus e rotavírus) de contaminação. Vinte e sete amostras de água provenientes de poços artesianos, poços cavados e fontes foram coletadas entre fevereiro e abril de 2011. A determinação do número mais provável (NMP) de E. coli e coliformes totais em 100 mL foi realizada pelo método dos tubos múltiplos. Os marcadores virais foram analisados em alíquotas de 500 mL por reação em cadeia da polimerase (PCR). Coliformes totais foram encontrados em 66,6% das amostras. A contaminação encontrada para coliformes termotolerantes foi de 25,9%, 40,7% para adenovírus e 22,2% para rotavírus. Nenhuma amostra apresentou enterovírus. A presença de coliformes e de vírus nas amostras indica contaminação fecal da água subterrânea no local de estudo, situado na porção aflorante do Aquífero Guarani e alerta para a necessidade de precaução quanto ao uso destes mananciais para abastecimento público, captação individual e cultivo de hortaliças.Palavras-chave: adenovírus, Escherichia coli, enterovírus, rotavírus. Viral and bacterial contamination in groundwater from the GuaraníAquifer's recharge area, Ivoti municipality, RS ABSTRACTThe water supply of the municipality of Ivoti comes solely from an underground source. The potability of groundwater in the municipality was evaluated by analyzing bacterial (Escherichia coli and total coliforms) and viral (adenovirus, enterovirus and rotavirus) markers for contamination. Twenty-seven water samples from artesian wells, dug wells and springs were collected between February and April 2011. The determination of the most probable number
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