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.

Occurrence of human sapoviruses in wastewater and stool samples in Rio De Janeiro, Brazil.

AIM: To determine the prevalence and molecular epidemiology of human sapovirus (SaV) in both wastewater and stool samples in a 3-year (2012-2014) surveillance study performed in Rio de Janeiro State, Brazil. METHODS AND RESULTS: A total of 156 wastewater and 341 stool samples were analysed using quantitative real-time PCR. SaV was detected in 3·5% (12/341) in stool samples with virus load concentrations ranging from 10(4) to 10(9) genome copies per gram (gc g(-1) ), and in 33·0% (51/156) wastewater samples, with range concentration varying from 10(4) to 10(6)  gc l(-1) . Partial genome sequencing of wastewater and stool samples revealed the circulation of genotypes GI.1, GI.2, GI.6, GII.1 and GV.1. CONCLUSION: This study demonstrated the prevalence of human SaV in acute gastroenteritis (AGE) cases and revealed, for the first time, the environmental dissemination of those viruses in Brazil. SIGNIFICANCE AND IMPACT OF THE STUDY: SaV diagnosis should be considered in hospitalized children with AGE and the higher positive rate detection in environmental samples suggests that SaV infection could be underestimated or associated with asymptomatic cases.

Rotavirus seasonality in urban sewage from Argentina: effect of meteorological variables on the viral load and the genetic diversity.

In Argentina, the rotavirus disease exhibits seasonal variations, being most prevalent in the fall and winter months. To deepen the understanding of rotavirus seasonality in our community, the influence of meteorological factors on the rotavirus load and the genetic diversity in urban raw sewage from Córdoba city, Argentina were evaluated. Wastewater samples were collected monthly during a three-year study period and viral particles were concentrated by polyethylene glycol precipitation. RT-nested PCR was applied for rotavirus detection, and VP7/VP4 characterization and real-time PCR for rotavirus quantification. Both molecular techniques showed relatively similar sensitivity rates and revealed rotavirus presence in urban wastewater in cold and warm seasons, indicating its circulation in the local community all year round. However, a slight trend for rotavirus circulation was noted by real-time PCR in the fall and winter seasons, showing a significantly higher peak of rotavirus concentration at mean temperatures lower than 18°C and also higher, although not statistically different during drier weather. VP7 and VP4 gene characterization showed that G1 and P[8] genotypes were dominant, and temporal variations in genotype distribution were not observed. Rotavirus spread is complex and our results point out that weather factors alone cannot explain the seasonal quantitative pattern of the rotavirus disease. Therefore, alternative transmission routes, changes in human behavior and susceptibility, and the stability and survivability of the virus might all together contribute to the seasonality of rotavirus. The results obtained here provide evidence regarding the dynamics of rotavirus circulation and maintenance in Argentina.

Gastroenteric virus dissemination and influence of rainfall events in urban beaches in Brazil.

AIMS: This study was conducted to assess rotavirus A (RV-A), genogroup II (GII) norovirus (NoV), and human adenovirus (HAdV) dissemination in recreational water in an urban beach located in the city of Rio de Janeiro and their persistence during rainfall events. METHODS AND RESULTS: Viruses, including bacteriophage (PP7), used as internal control, were concentrated, reverse transcribed and quantified by a low-cost method based on organic flocculation with skimmed milk coupled with quantitative polymerase chain reaction protocols. The analysis of 74 superficial water samples obtained during 6 months of monitoring detected HAdV (66%), RV-A (37%) and GII NoV (14%), with a mean viral load of 4·1 log10 genome copies l(-1) (g.c. l(-1) ), 4·3 log10 g.c l(-1) and 3·8 log10 g.c. l(-1) , respectively. Investigation of those viruses during two rainfall events showed a longer permanence after rainfall events compared with bacterial indicators. CONCLUSIONS: The results highlight the need for further monitoring using viral parameters to determine the microbiological quality of recreational waters to allow bath in these waters, especially during rainy events. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides data on virus contamination in recreational waters on tourist beaches frequented throughout the year, emphasizing the importance of viral parameters for assessing microbiological quality of water, as well as the potential risk of waterborne infections.

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.

Acute gastroenteritis cases associated with noroviruses infection in the state of Rio de Janeiro.

In March 2005, the Epidemiological Surveillance Service of Resende, municipality of the Middle Paraiba Valley, State of Rio de Janeiro, reported a sudden spontaneous occurrence of acute gastroenteritis cases in children in a public day care center. Further, between May and June 2005, gastroenteritis outbreaks or sporadic cases of gastroenteritis were reported in two other municipalities, Piraí and Rio Claro, also located in the Middle Paraiba Valley. From March to June 2005, 50 fecal samples were collected in this region and those samples were tested for the presence of bacteria and other parasites and were demonstrated to be negative. Polyacrylamide gel electrophoresis and an enzyme immunoassay were performed for adenovirus and rotavirus detection and reverse transcription-polymerase chain reaction was performed to investigate the presence of norovirus (NoV) and astrovirus. In addition, a quantitative TaqMan real time PCR for NoV was performed for quantification of viral DNA in order to compare the results with those obtained by conventional RT-PCR. NoV was detected in 33 out of 50 (66%) samples, and a 100% correlation between both methodologies was obtained. These results are demonstrating that NoV was the etiological agent responsible for those acute gastroenteritis cases. Partial nucleotide sequence analysis of the capsid gene revealed that the circulating strain was NoV GII/4 confirming the worldwide distribution of this genotype. The results highlight the role of NoV as a main viral agent responsible for gastroenteritis cases in children and adults both in outbreaks as well as in sporadic cases of acute gastroenteritis.