Bioaccumulation of animal adenoviruses in the pink shrimp

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..

Bioaccumulation of animal adenoviruses in the pink shrimp

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..(AU)

Detection of enteric viruses in activated sludge by feasible concentration methods

Human enteric viruses are responsible to cause several diseases, including gastroenteritis and hepatitis, and can be present in high amounts in sewage sludge. This study compared virus recovery efficiency of two feasible concentration methods used for detecting human adenovirus (HAdV), rotavirus species A (RV-A), norovirus genogroup II (NoV GII) and hepatitis A virus (HAV) in sewage sludge from an activated sludge process. Twelve sewage sludge samples were collected bi-monthly from January to July, 2011. Ultracentrifugation was compared with a simplified protocol based on beef extract elution for recovering enteric viruses. Viruses were quantified by quantitative real-time PCR assays and virus recovery efficiency and limits of detection were determined. Methods showed mean recovery rates lower than 7.5%, presenting critical limits of detection (higher than 102 103 genome copies -GC L-1 for all viruses analyzed). Nevertheless, HAdV were detected in 90% of the analyzed sewage sludge samples (range: 1.8 x 104 to 1.1 x 105 GC L-1), followed by RV-A and NoV (both in 50%) and HAV (8%). Results suggesting that activated sludge is contaminated with high viral loads and HAdV are widely disseminated in these samples. The low virus recovery rates achieved, especially for HAV, indicate that other feasible concentration methods could be developed to improve virus recovery efficiency in these environmental matrices.

Detection of enteric viruses in activated sludge by feasible concentration methods

Human enteric viruses are responsible to cause several diseases, including gastroenteritis and hepatitis, and can be present in high amounts in sewage sludge. This study compared virus recovery efficiency of two feasible concentration methods used for detecting human adenovirus (HAdV), rotavirus species A (RV-A), norovirus genogroup II (NoV GII) and hepatitis A virus (HAV) in sewage sludge from an activated sludge process. Twelve sewage sludge samples were collected bi-monthly from January to July, 2011. Ultracentrifugation was compared with a simplified protocol based on beef extract elution for recovering enteric viruses. Viruses were quantified by quantitative real-time PCR assays and virus recovery efficiency and limits of detection were determined. Methods showed mean recovery rates lower than 7.5%, presenting critical limits of detection (higher than 102 103 genome copies -GC L-1 for all viruses analyzed). Nevertheless, HAdV were detected in 90% of the analyzed sewage sludge samples (range: 1.8 x 104 to 1.1 x 105 GC L-1), followed by RV-A and NoV (both in 50%) and HAV (8%). Results suggesting that activated sludge is contaminated with high viral loads and HAdV are widely disseminated in these samples. The low virus recovery rates achieved, especially for HAV, indicate that other feasible concentration methods could be developed to improve virus recovery efficiency in these environmental matrices.(AU)

Detection of rotavirus from pig livestock wastewater of São Paulo State, Brazil / Detecção de rotavírus em efluentes de criações de suínos no estado de São Paulo, Brasil

Rotavirus is one of the most important viral agents of gastroenteritis among child and animals from different species. It's high environmental resistance and the fecal-oral way of transmission makes this virus likely to be transmitted by wastewater. This study seeks to detect the wastewater elimination and circulation of group A rotavirus in low technified pig farms from São Paulo State, Brazil. A total of 25 samples, including piglet feces with diarrhea and untreated wastewater samples, from 7 different farms, were submitted in a parallel screening scheme of rotavirus infection through polyacrilamide gel electrophoresis (PAGE) and ELISA, which the positive samples were further confirmed by RT-PCR (reverse-transcription polimerase chain reaction). The PAGE revealed only one positive sample (1/25 or 4%) from feces, while by ELISA, 6 (6/25 or 24%) samples were positive, which 4 were from feces and 2 from wastewater. The RT-PCR confirmed all positive PAGE and ELISA results. Therefore, the rotavirus was found in 3 of 7 (42.86%) researched farms, which in 2 of these were detected both in animals and wastewater and one were found virus only in fecal samples. In view of these results, there was rotavirus detection from untreated pig farm wastewater, posing as a risk of spreading for humans and animals, implying the need of assuring microbiological and environmental safety measures with this material.

Os rotavírus são um dos principais agentes virais envolvidos na ocorrência de gastroenterites em crianças e em animais de diferentes espécies. Sua elevada resistência ambiental aliada à via de transmissão fecal-oral torna-o um agente propício de se propagar pela água, principalmente nos efluentes. O objetivo deste estudo foi o de se detectar a circulação e eliminação de rotavírus em criações de suínos de baixa tecnificação do Estado de São Paulo, Brasil. Um total de 25 amostras, incluindo fezes de leitões com diarréia e efluentes não tratados, de 7 diferentes propriedades, foram testadas em paralelo para detecção do rotavírus através da eletroforese em gel de poliacrilamida (PAGE) e ELISA, sendo as positivas confirmadas por RT-PCR (transcrição reversa - reação em cadeia pela polimerase). A PAGE evidenciou apenas uma amostra positiva (1/25 ou 4%) proveniente de material fecal, enquanto que pela ELISA, 6 (6/25 ou 24%) amostras positivas, das quais 4 de material fecal e 2 de efluentes. A RT-PCR confirmou todos os resultados positivos de PAGE e ELISA. Portanto, os rotavírus foram encontrados em 3 de 7 (42.86%) das criações pesquisadas, das quais em duas destas, o vírus foi detectado tanto no efluente quanto nos animais. Em face destes resultados, houve a detecção de rotavírus nos efluentes não tratados de criações de suínos, constituindo um risco para a disseminação do agente para humanos e animais, implicando na necessidade de assegurarem-se medidas de segurança ambiental e microbiológica deste material.