Distribution and genetic diversity of the human polyomaviruses JC and BK in surface water and sewage treatment plant during 2009 in Porto Alegre, Southern Brazil / Distribuição e diversidade genética dos poliomavírus humanos JC e BK em águas superficiais e de estação de tratamento de esgoto durante 2009 em Porto Alegre, sul do Brasil

Abstract The human polyomaviruses JC and BK (JCPyV and BKPyV) are ubiquitous, species-specific viruses that belong to the family Polyomaviridae. These viruses are known to be excreted in human urine, and they are potential indicators of human wastewater contamination. In order to assess the distribution of both JCPyV and BKPyV in urban water samples collected from a sewage treatment plant (STP) and from a canalized water stream of Porto Alegre, Brazil, two nested-PCR assays were optimized and applied to the samples collected. The amplicons obtained were submitted to sequencing, and the sequences were analyzed with sequences of human polyomaviruses previously deposited in GenBank. Twelve out of 30 water samples (40%) were JCPyV positive, whereas six samples (20%) were BKPyV positive. The sequencing results confirmed the presence of JCPyV subtypes 1 and 3, whereas only BKPyV Ia and Ib were found. This study shows for the first time the presence of human polyomaviruses in surface water and in samples collected in a sewage treatment plant in southern Brazil.

Resumo Os poliomavírus humanos JC e BK (JCPyV e BKPyV) são virus ubíquos, espécie-específicos, pertencentes à família Polyomaviridae. Estes vírus são conhecidos por serem excretados pela urina humana, sendo considerados potenciais indicadores de contaminação por águas residuais urbanas. Buscando acessar a distribuição de JCPyV e BKPyV em amostras de águas coletadas de uma estação de tratamento de esgoto e de um arroio canalizado de Porto Alegre, Brasil, duas técnicas de nested-PCR foram otimizadas e aplicadas às amostras coletadas. Os amplificados obtidos foram submetidos ao sequenciamento e suas sequências analisadas com base em sequências de poliomavírus humanos previamente depositadas no GenBank. Doze de 30 amostras de água (40%) foram positivas para JCPyV, enquanto 6 amostras (20%) foram positivas para BKPyV. Os resultados do sequenciamento confirmaram a presença dos subtipos 1 e 3 de JCPyV, enquanto apenas os BKPyV Ia e Ib foram encontrados. Este estudo demonstra pela primeira vez a presença de poliomavírus humanos em águas superficiais e em amostras coletadas em uma estação de tratamento de esgoto na região sul do Brasil.

Distribution and genetic diversity of the human polyomaviruses JC and BK in surface water and sewage treatment plant during 2009 in Porto Alegre, Southern Brazil.

The human polyomaviruses JC and BK (JCPyV and BKPyV) are ubiquitous, species-specific viruses that belong to the family Polyomaviridae. These viruses are known to be excreted in human urine, and they are potential indicators of human wastewater contamination. In order to assess the distribution of both JCPyV and BKPyV in urban water samples collected from a sewage treatment plant (STP) and from a canalized water stream of Porto Alegre, Brazil, two nested-PCR assays were optimized and applied to the samples collected. The amplicons obtained were submitted to sequencing, and the sequences were analyzed with sequences of human polyomaviruses previously deposited in GenBank. Twelve out of 30 water samples (40%) were JCPyV positive, whereas six samples (20%) were BKPyV positive. The sequencing results confirmed the presence of JCPyV subtypes 1 and 3, whereas only BKPyV Ia and Ib were found. This study shows for the first time the presence of human polyomaviruses in surface water and in samples collected in a sewage treatment plant in southern Brazil.

Human adenovirus (HAdV), human enterovirus (hEV), and genogroup A rotavirus (GARV) in tap water in southern Brazil.

The effects of viral gastroenteritis are more devastating in children than in any other age category. Thus, children exposed to the consumption of low quality water are at an increased risk of infection, especially in regions where sanitation is inadequate. The present study aimed to provide a survey of the occurrence of representative enteric viruses: human adenovirus (HAdV), human enteroviruses (hEV), and genogroup A rotavirus (GARV) in tap water samples collected in public schools located at six municipalities of Rio Grande do Sul, southern Brazil. Seventy-three schools were included in the study and tap water samples were analyzed by conventional PCR for the presence of HAdV, hEV, and GARV genomes. hEV showed the highest detection rate (27.4%), followed by HAdV (23.3%), and GARV (16.4%). New approaches to water monitoring should be considered to promote a better water quality and reduce the risk of waterborne diseases, especially considering drinking water to be served to vulnerable individuals.

Assessment of enteric viruses in a sewage treatment plant located in Porto Alegre, southern Brazil / Avaliação de vírus entéricos em uma estação de tratamento de esgoto localizada em Porto Alegre, sul do Brasil

In order to verify the microbial quality of the influents and effluents of one STP from southern Brazil, an eight-month survey was conducted to examine the presence of total and fecal coliforms and of adenovirus (HAdV), enterovirus (EV), genogroup A rotaviruses (GARV) and Torque teno virus (TTV), in treated effluent samples from São João/Navegantes STP, Porto Alegre (Brazil). A total of 16 samples were collected, eight of influent (raw sewage, prior to treatment), and the other eight of the effluent (post-treatment sewage). Total and fecal coliform levels ranging from 3.6 × 10(4) to 4.4 × 10(7) MPN/100 mL and 2.9 × 10³ to 1.7 × 10(7) MPN/100 mL, were detected in all samples. In raw sewage, HAdV (25%) and GARV (28.6%) viral genomes were detected. The analysis of effluent samples revealed the presence of HAdV (50%), EV (37.5%), and TTV (12.5%) genomic fragments. All samples, regardless of the month analysed, presented detection of a least one virus genus, except for in April. Higher virus detection rates were observed in treated sewage samples (62.5%), and in 80% of them (effluent positive samples) HAdV was detected. Results showed that improvements in sewage monitoring and treatment processes are necessary to reduce the viral and bacterial load on the environment in southern Brazil. To the knowledge of the authors, this is the first study showing the monitoring of viral genomes in influent and effluent samples from a STP located in Porto Alegre (Rio Grande do Sul, Brazil), southern Brazil.

Com o intuito de verificar a qualidade microbiológica de afluentes e efluentes de uma estação de tratamento de esgoto (ETE), um monitoramento de oito meses foi realizado para examinar a presença de coliformes totais e fecais, e de adenovírus (HAdV), enterovírus (EV), rotavírus do genogrupo A (GARV) e torque teno vírus (TTV), em amostras de esgoto tratado da ETE São João/Navegantes, em Porto Alegre-RS, Brasil. Um total de 16 amostras foi coletado, sendo oito de afluente (esgoto bruto, anterior ao tratamento) e oito de efluente (esgoto tratado). Os níveis de coliformes totais e fecais variaram entre 3,6 × 10(4) e 4,4 × 10(7) MPN/100 mL e 2,9 × 10³ e 1,7 × 10(7) MPN/100 mL, respectivamente, tendo sido estes detectados em todas as amostras. No esgoto bruto, foram detectados os genomas virais de HAdV (25%) e GARV (28,6%). A análise das amostras de efluente revelou a presença de fragmentos genômicos de HAdV (50%), EV (37,5%) e TTV (12,5%). Todas as amostras, independentemente do mês analisado, possibilitaram a detecção de pelo menos um gênero viral, exceto no mês de abril. Altas taxas de detecção viral foram observadas em amostras de esgoto tratado (62,5%), sendo que o HAdV foi detectado em 80% dessas amostras de efluente positivas. Os resultados mostram que aprimoramentos no processo de tratamento e monitoramento do esgoto são necessários para reduzir a carga viral e bacteriológica no ambiente do Sul do Brasil. Ao conhecimento dos autores, este é o primeiro estudo de monitoramento de genomas virais em amostras de afluente e efluente de uma ETE localizada em Porto Alegre-Rio Grande do Sul, Brasil.

First description of Adenovirus, Enterovirus, Rotavirus and Torque teno virus in water samples collected from the Arroio Dilúvio, Porto Alegre, Brazil.

Adenovirus (AdV), enterovirus (EV), genogroup A rotaviruses (GARV) and Torque teno virus (TTV) are non-enveloped viral agents excreted in feces and so may contaminate water bodies. In the present study, the molecular detection of these viruses was performed in samples of surface water collected from the Arroio Dilúvio, a waterstream that crosses the city of Porto Alegre, RS, Brazil, receiving great volumes of non-treated sewage from a large urban area. Sampling was performed during 2009, in three different occasions (January, April and September). The highest detection rate was observed for EV (64.28%), followed by TTV (28.57%) and AdV (21.43%). Rotaviruses were not detected. More than on kind of tested virus was detected in five (35. 71%) of 14 samples. January was the month with the highest viral detection rate, being all samples, collected in this month, positive for at least one group of tested virus. The correlation between the detection of these different viral agents and environmental factors is discussed. To the knowledge of the authors, this is the first description of viral genomes in water samples taken from the Arroio Dilúvio, Porto Alegre (Brazil).

First description of Adenovirus, Enterovirus, Rotavirus and Torque teno virus in water samples collected from the Arroio Dilúvio, Porto Alegre, Brazil / Primeira descrição de Adenovírus, Enterovírus, Rotavírus e Torque teno vírus em amostras de água coletadas do Arroio Dilúvio, Porto Alegre, Brasil

Adenovirus (AdV), enterovirus (EV), genogroup A rotaviruses (GARV) and Torque teno virus (TTV) are non-enveloped viral agents excreted in feces and so may contaminate water bodies. In the present study, the molecular detection of these viruses was performed in samples of surface water collected from the Arroio Dilúvio, a waterstream that crosses the city of Porto Alegre, RS, Brazil, receiving great volumes of non-treated sewage from a large urban area. Sampling was performed during 2009, in three different occasions (January, April and September). The highest detection rate was observed for EV (64.28%), followed by TTV (28.57%) and AdV (21.43%). Rotaviruses were not detected. More than on kind of tested virus was detected in five (35. 71%) of 14 samples. January was the month with the highest viral detection rate, being all samples, collected in this month, positive for at least one group of tested virus. The correlation between the detection of these different viral agents and environmental factors is discussed. To the knowledge of the authors, this is the first description of viral genomes in water samples taken from the Arroio Dilúvio, Porto Alegre (Brazil).

Adenovírus (AdV), enterovírus (EV), rotavírus (GARV) e Torque teno vírus (TTV) são vírus não envelopados, excretados nas fezes, podendo, assim, contaminar corpos hídricos. No presente estudo, a detecção molecular desses agentes foi realizada em amostras de águas superficiais provenientes do Arroio Dilúvio, o qual cruza a cidade de Porto Alegre-RS, Brasil. As amostras foram coletadas em três meses diferentes (janeiro, abril e setembro) do ano de 2009. A maior taxa de detecção viral foi observada para EV (64,28%), seguida por TTV (28,57%) e AdV (21,43%). Rotavírus não foi detectado. Foi verificada presença simultânea de dois grupos virais em cinco (35,71%) das 14 amostras analisadas. Janeiro foi o mês com a maior taxa de detecção viral, sendo todas as amostras, coletadas nesse mês, positivas para, no mínimo, um grupo viral em estudo. A correlação entre a detecção desses diferentes agentes virais e os fatores ambientais é discutida. Conforme conhecimento dos autores, essa é a primeira descrição de genomas virais em amostras de água provenientes do Arroio Dilúvio, Porto Alegre, Brasil.

Assessment of enteric viruses in a sewage treatment plant located in Porto Alegre, southern Brazil.

In order to verify the microbial quality of the influents and effluents of one STP from southern Brazil, an eight-month survey was conducted to examine the presence of total and fecal coliforms and of adenovirus (HAdV), enterovirus (EV), genogroup A rotaviruses (GARV) and Torque teno virus (TTV), in treated effluent samples from São João/Navegantes STP, Porto Alegre (Brazil). A total of 16 samples were collected, eight of influent (raw sewage, prior to treatment), and the other eight of the effluent (post-treatment sewage). Total and fecal coliform levels ranging from 3.6 × 10(4) to 4.4 × 10(7) MPN/100 mL and 2.9 × 10(3) to 1.7 × 10(7) MPN/100 mL, were detected in all samples. In raw sewage, HAdV (25%) and GARV (28.6%) viral genomes were detected. The analysis of effluent samples revealed the presence of HAdV (50%), EV (37.5%), and TTV (12.5%) genomic fragments. All samples, regardless of the month analysed, presented detection of a least one virus genus, except for in April. Higher virus detection rates were observed in treated sewage samples (62.5%), and in 80% of them (effluent positive samples) HAdV was detected. Results showed that improvements in sewage monitoring and treatment processes are necessary to reduce the viral and bacterial load on the environment in southern Brazil. To the knowledge of the authors, this is the first study showing the monitoring of viral genomes in influent and effluent samples from a STP located in Porto Alegre (Rio Grande do Sul, Brazil), southern Brazil.

Enterovirus infections and type 1 diabetes mellitus: is there any relationship?

Several health organizations have classified diabetes mellitus, a metabolic syndrome, as the epidemic of the century, since it affects millions of people worldwide and is one of the top ten causes of death. Type 1 diabetes is considered to be an autoimmune disease, in which autoaggressive T cells infiltrate the islets of Langerhans in the pancreas, leading to the destruction of insulin producing beta cells. The risk of the disease is modulated by genetic factors, mainly genes coding for human leukocyte antigens (HLA). However, the incidence of this disease has increased significantly during the recent decades, which cannot be explained only by genetic factors. Environmental perturbations have also been associated to the development of diabetes. Among these factors, viral triggers have been implicated; particularly enteroviruses, which have been associated to the induction of the disease. Supporting the hypothesis, numerous lines of evidence coming from mouse models and patients with this type of diabetes have shown the association. The present review aims to provide some understanding of how type 1 diabetes occurs and the possible role of enterovirus in this pathology.

HbA(1c) measurement for the diagnosis of diabetes: is it enough?

AIM: To analyse the performance of HbA(1c) in diagnosing Type 2 diabetes based on fasting plasma glucose and/or 2-h plasma glucose measurements after a 75-g oral glucose tolerance test. METHODS: This is a study of diagnostic test accuracy in individuals referred to the Clinical Pathology Department for oral glucose tolerance testing. After fasting overnight, HbA(1c), fasting plasma glucose and 2-h plasma glucose were measured. The receiver operating characteristic curve was used to evaluate the diagnostic performance of HbA(1c). RESULTS: Four hundred and ninety-eight subjects (195 male, mean age 56 years) were enrolled and 115 (23.1%) were diagnosed with diabetes according to glucose-based methods and only 56 (11.2%) individuals were identified by HbA(1c) ≥ 6.5% (48 mmol/mol) (sensitivity 20.9%, specificity 95.3%). There is poor agreement between the newly recommended criterion and the current glucose-based diagnostic criteria (κ = 0.217; P < 0.001), probably because the diagnostic methods identify different populations of patients. Adding a glucose-based method into an algorithm, as proposed by the UK Department of Health, improved HbA(1c) performance. CONCLUSIONS: HbA(1c) ≥ 6.5% (48 mmol/mol) showed limited sensitivity to diabetes diagnosis, although with high specificity. The results suggest that this cut-off point would not be enough to diagnose diabetes. Its use as the sole diabetes diagnostic test should be interpreted with caution to assure the correct classification of diabetic individuals.