Norovírus em alimentos

Introduction: Noroviruses (NoV) are important causative agents of foodborne gastroenteritis outbreaks associated with the consumption of fruits, leafy vegetables, bivalve molluscs and delicatessen foods. The establishment of laboratory surveillance networks in different continents has demonstrated increased epidemiological importance of those viruses. In Brazil, the NoV infection is considered an important public health issue with socioeconomic burden, but the investigation of these viruses in foodborne outbreaks is still restricted to research laboratories. NoV infections have become more known especially with the consolidation of the cruise ship market in the country since 2004. Objective: This study aims to present advances related to NoV research in foods, highlighting features of this pathogen and strategies for its detection in these matrices. Method: An integrative review, collecting scientific articles with the objective of dealing with the main aspects of NoV, was carried out. Results: A broad literature review was performed, describing the main results in the literature and discussing aspects such as foodborne diseases, viruses as food contaminants, stability and disinfection, foodborne outbreaks associated with NoV, food associated with NoV contamination, NoV concentration and detection methods in food, risk assessment studies and prevention and control. Conclusions: records of foodborne outbreaks associated with NoV and the increasing genetic diversity of these viruses reinforce the need for laboratory and epidemiological surveillance, especially in developing countries, such as Brazil.Introdução: Os norovírus (NoV) são importantes agentes causadores de gastroenterite de origem alimentar, com surtos associados ao consumo de frutas, vegetais folhosos, moluscos bivalves e alimentos de delicatessen. O aumento da importância epidemiológica destes vírus tem sido demonstrado pelo estabelecimento de redes laboratoriais de vigilância em diversos continentes. As infecções por NoV se tornaram mais conhecidas especialmente com a consolidação do mercado de navios de cruzeiros no país a partir de 2004. Objetivo: Este estudo tem como objetivo apresentar avanços relacionados à pesquisa de NoV em alimentos, destacando características deste patógeno e estratégias para sua detecção nestas matrizes. Método: Foi realizada uma revisão integrativa, pelo levantamento de artigos científicos com o objetivo de tratar dos principais aspectos de NoV. Resultados: Foi realizada uma ampla revisão da literatura, com a descrição dos principais resultados presentes na literatura consultada e a discussão de aspectos como doenças transmitidas por alimentos (DTA), vírus como contaminantes de alimentos, estabilidade e desinfecção, surtos de origem alimentar associados aos NoV, alimentos associados à contaminação por NoV, métodos de concentração e detecção de NoV em alimentos, estudos de avaliação de risco e prevenção e controle. Conclusões: Os registros de envolvimento de NoV em surtos de origem alimentar e a crescente diversidade genética destes vírus reforçam a necessidade de vigilância laboratorial e epidemiológica sobretudo nos países em desenvolvimento, como o Brasil

Norovírus em alimentos

Introduction: Noroviruses (NoV) are important causative agents of foodborne gastroenteritis outbreaks associated with the consumption of fruits, leafy vegetables, bivalve molluscs and delicatessen foods. The establishment of laboratory surveillance networks in different continents has demonstrated increased epidemiological importance of those viruses. In Brazil, the NoV infection is considered an important public health issue with socioeconomic burden, but the investigation of these viruses in foodborne outbreaks is still restricted to research laboratories. NoV infections have become more known especially with the consolidation of the cruise ship market in the country since 2004. Objective: This study aims to present advances related to NoV research in foods, highlighting features of this pathogen and strategies for its detection in these matrices. Method: An integrative review, collecting scientific articles with the objective of dealing with the main aspects of NoV, was carried out. Results: A broad literature review was performed, describing the main results in the literature and discussing aspects such as foodborne diseases, viruses as food contaminants, stability and disinfection, foodborne outbreaks associated with NoV, food associated with NoV contamination, NoV concentration and detection methods in food, risk assessment studies and prevention and control. Conclusions: records of foodborne outbreaks associated with NoV and the increasing genetic diversity of these viruses reinforce the need for laboratory and epidemiological surveillance, especially in developing countries, such as Brazil.Introdução: Os norovírus (NoV) são importantes agentes causadores de gastroenterite de origem alimentar, com surtos associados ao consumo de frutas, vegetais folhosos, moluscos bivalves e alimentos de delicatessen. O aumento da importância epidemiológica destes vírus tem sido demonstrado pelo estabelecimento de redes laboratoriais de vigilância em diversos continentes. As infecções por NoV se tornaram mais conhecidas especialmente com a consolidação do mercado de navios de cruzeiros no país a partir de 2004. Objetivo: Este estudo tem como objetivo apresentar avanços relacionados à pesquisa de NoV em alimentos, destacando características deste patógeno e estratégias para sua detecção nestas matrizes. Método: Foi realizada uma revisão integrativa, pelo levantamento de artigos científicos com o objetivo de tratar dos principais aspectos de NoV. Resultados: Foi realizada uma ampla revisão da literatura, com a descrição dos principais resultados presentes na literatura consultada e a discussão de aspectos como doenças transmitidas por alimentos (DTA), vírus como contaminantes de alimentos, estabilidade e desinfecção, surtos de origem alimentar associados aos NoV, alimentos associados à contaminação por NoV, métodos de concentração e detecção de NoV em alimentos, estudos de avaliação de risco e prevenção e controle. Conclusões: Os registros de envolvimento de NoV em surtos de origem alimentar e a crescente diversidade genética destes vírus reforçam a necessidade de vigilância laboratorial e epidemiológica sobretudo nos países em desenvolvimento, como o Brasil

Detection and quantification of classic and emerging viruses by skimmed-milk flocculation and PCR in river water from two geographical areas

Molecular techniques and virus concentration methods have shown that previously unknown viruses are shed by humans and animals, and may be transmitted by sewage-contaminated water. In the present study, river water from urban areas in Barcelona, Spain and Rio de Janeiro, Brazil, were analyzed to evaluate the dissemination of human viruses, while simultaneously optimizing and validating a low-cost concentration method for virus quantification in fresh water. The following three viral groups were analyzed. (i) Recently described viruses: klassevirus (KV), asfarvirus-like virus (ASFLV), and the polyomaviruses Merkel cell, KI and WU (MCPyV/KIPyV/WUPyV). (ii) Gastroenteritis agents: noroviruses (NoV) and rotaviruses (RV). (iii) Human fecal viral indicators in water: human adenoviruses (HAdV) and JC polyomaviruses (JCPyV). Virus detection was based on nested and quantitative PCR assays. Nested PCR assays were developed for KV and ASFLV. The method applied for virus concentration in water samples was a one-step procedure based on a skimmed milk flocculation procedure described previously for seawater. Using spiked river water samples, inter- and intra-laboratory assays showed a viral recovery rate of about 50% for HAdV, JCPyV, NoV and RV with a coefficient of variation ≤ 50%. HAdV and JCPyV were detected in 100% of the river samples from Barcelona and Rio de Janeiro. Moreover, NoV GGII was detected in 100% and MCPyV in 50% of the samples from Barcelona, whereas none of the other viruses analyzed were detected. NoV GGII was detected in 33%, KV in 33%, ASFLV in 17% and MCPyV in 50% of the samples from Rio de Janeiro, whereas KIPyV and WUPyV were not detected. RV were only tested for in Rio de Janeiro and resulted positive in 67% of the samples. The procedure applied here to river water represents a useful, straightforward and cost-effective method that could be applied in routine water quality testing.  The results of the assays expand our understanding of the global distribution of the viral pathogens studied here and their persistence in the environment. Fil: Calgua, B.. Universidad de Barcelona; España;Fil: Fumian, T.. Ministerio de Salud de Brasil. Fundacion Oswaldo Cruz; Brasil;Fil: Rusinol, M.. Universidad de Barcelona; España;Fil: Rodríguez Manzano, J.. Universidad de Barcelona; España;Fil: Mbayed, Viviana Andrea. Universidad de Buenos Aires. Facultad de Farmacia y Bioquímica. Departamento de Microbiología, Inmunología y Biotecnología. Cátedra de Virología; Argentina. Consejo Nacional de Investigaciones Científicas y Técnicas. Oficina de Coordinación Administrativa Houssay; ArgentinaFil: Bofill Mas, S.. Universidad de Barcelona; España;Fil: Miagostovich, M.. Ministerio de Salud de Brasil. Fundacion Oswaldo Cruz; Brasil;Fil: Girones, R.. Universidad de Barcelona; España

Dengue Virus Type 3, Brazil, 2002

An explosive epidemic of DENV-3 in 2002 was the most severe dengue epidemic reported in Brazil since dengue viruses were introduced

Human adenovirus in municipal solid waste leachate and quantitative risk assessment of gastrointestinal illness to waste collectors

Leachate is a variable effluent from waste management systems generated during waste collection and on landfills. Twenty-two leachate samples from waste collection trucks and a landfill were collected from March to December 2019 in the municipality of Rio de Janeiro (Brazil) and were analyzed for Human Adenovirus (HAdV), bacterial indicators and physico-chemical parameters. For viral analysis, samples were concentrated by ultra centrifugation and processed for molecular analysis using QIAamp Fast DNA Stool mini kit (R) for DNA extraction followed by nested-PCR and qPCR/PMA-qPCR TaqMan (R) system. HAdV was detected by nested-PCR in 100% (9/ 9) and 83.33% (12/13) of the truck and landfill leachate samples, respectively. Viral concentrations ranged from 8.31 x 10(1) to 6.68 x 107 genomic copies per 100 ml by qPCR and PMA-qPCR. HAdV species A, B, C, and F were characterized using nucleotide sequencing. HAdV were isolated in A549 culture cells in 100% (9/9) and 46.2% (6/13) from truck and landfill leachate samples, respectively. Regardless of the detection methods, HAdV concentration was predicted by the quantity of total suspended solids. A quantitative microbial risk assessment was performed to measure the probability of gastrointestinal (GI) illness attributable to inadvertent oral ingestion of truck leachate, revealing the higher probability of disease for the direct splashing into the oral cavity (58%) than for the gloved hand-to-mouth (33%). In a scenario where waste collectors do not wear gloves as protective personal equipment, the risk increases to 67%. This is the first study revealing infectious HAdV in solid waste leachate and indicates a potential health risk for waste collectors

Recovery of Norovirus from lettuce (Lactuca sativa) using an adsorption —elution method with a negatively charged membrane: comparison of two elution buffers

Phosphate saline buffer (PBS) and glycine buffer (GB) were evaluated as elution buffers in an adsorption-elution method using a negatively charged membrane associated with quantitative polymerase chain reaction (qPCR) and seminested PCR for detection of Norovirus genogroup II (NoV GII) from lettuce. In this methodology, PP7 bacterio-phage was used as a virus sample for process control. The qPCR showed more sensitivity than semi-nested PCR for NoV GII detection. The recovery efficiency, using PBS and GB, ranged from 24.72 to 60.78% and 19.48 to 137.26% for NoV GII, and from 0.01 to 0.15% and 0.13 to 6.04% for PP7 bacteriophage, respectively. Elution with GB was more effi-cient for PP7 bacteriophage recovery (p = 0.03), but no difference was seen for NoV GII (p = 0.57). The GB performed better than PBS as an eluent solution and can be consid-ered a methodological improvement.Título PT: Recuperação de Norovirus a partir de alface (Lactuca sativa) utilizando um método de adsorção-eluição com membrana negativamente carregada — comparação de dois tampões de eluiçãoSalina tamponada fosfatada (STF) e tampão glicina (TG) foram avaliados como eluentes em um método de adsorção-eluição, utilizando membrana carregada negativamente associada com reação quantitativa de polimerase em cadeia (qPCR) e semi-nested PCR, para detecção de Norovírus do genogrupo II (NoV GII) a partir de alface. Nesta metodologia, o bacteriófago PP7 foi utilizado como controle do processo. A qPCR apresentou maior sensibilidade do que o semi-nested PCR na detecção de NoV GII. A eficiência de recuperação, utilizando STF e TG, variou de 24,72 a 60,78% e de 19,48 a 137,26% para NoV GII e de 0,01 a 0,15% e de 0,13 a 6,04% para o bacteriófago PP7, respectivamente. A eluição com TG foi mais eficiente na recuperação do bacteriófago PP7 (p = 0,03) em-bora nenhuma diferença tenha sido observada para NoV GII (p = 0,57). O TG apresentou melhor desempenho que a STF como solução para eluição e pode ser considerada uma melhoria do método

Nearly complete genome sequence of a human norovirus GII.P17-GII.17 strain isolated from Brazil in 2015

Human noroviruses are the most common cause of nonbacterial acute gastroenteritis worldwide. We report here the nearly complete genome sequence (7,551 nucleotides) of a human norovirus GII.P17-GII.17 strain detected in July 2015 in the stool sample from an adult with acute gastroenteritis in Brazil