Influencia de la seropositividad contra adenovirus-36 sobre riesgo de obesidad y resistencia a la insulina en población infanto-juvenil de la Región de la Araucanía / Influence of adenovirus 36 seropositivity on the risk of obesity and insulin resistance in a population of children and adolescents from the Región de la Araucanía

La infección previa por el adenovirus-36 (Ad-36) se ha asociado con el proceso adipogénico y el control glicémico en modelos experimentales de cultivos celulares y animales. En humanos, la presencia de anticuerpos contra Ad-36 ha mostrado aumentar el riesgo de obesidad y, paradójicamente, mejorar el control glicémico en diferentes poblaciones. Se evaluó la influencia de la seropositividad contra Ad-36 sobre riesgo de obesidad, el perfil lipídico y glicémico en una población de niños en edad escolar. Métodos: Doscientos ocho individuos de entre 9 y 13 años se agruparon según estado nutricional como normopeso (IMC z-score de -1 a +1), con sobrepeso (IMC z-score de +1 a +2) y con obesidad (IMC z-score > +3). Se evaluaron medidas antropométricas, desarrollo puberal según Tanner y parámetros bioquímicos (perfil lipídico, glucemia e insulina) y la seropositividad contra Ad-36. Se determinó la resistencia a la insulina (RI) según criterio para la población infantil chilena. La seropositividad contra Ad-36 se determinó mediante ELISA. Resultados: Hubo una alta prevalencia de sobrepeso/obesidad en la población de estudio. La seropositividad contra Ad-36 fue del 5,4% en el grupo total, pero no se observó una asociación con el estado nutricional. No se encontró correlación entre la seropositividad contra Ad-36 y los parámetros del perfil lipídico. La insulina y la HOMA-RI fueron significativamente más bajas en el grupo Ad-36 (+) (p<0,001), no habiendo sido reportados casos de RI en el grupo Ad-36 (+) en nuestra población. Conclusiones: Nuestros resultados sugieren que la infección previa por el adenovirus-36 afecta la secreción de insulina y la resistencia a la insulina, como se ha descrito anteriormente, sin embargo, no se observa correlación con el desarrollo de la obesidad infantil en la población pediátrica del sur de Chile.

Previous infection with Adenovirus-36 (Ad-36) has been associated with adipogenic process and glycemic control in experimental models of cell culture and animals. In humans, the presence of antibodies against Ad-36 has been shown to increase the risk of obesity and, paradoxically, improve glycemic control in different populations. The influence of Ad-36 seropositivity on obesity risk, lipid and glycemic profile was evaluated in a population of school-age children. Methods: Two hundred eight individuals aged 9 to 13 years were grouped according to their nutritional status as normal weight (BMI z-score from -1 to +1), overweight (BMI z-score from +1 to +2) or obese (BMI z-score from -1 to +1). z-score > +3). Anthropometric measurements, pubertal development according to Tanner stage, biochemical parameters (lipid profile, glycemia and insulin) and seropositivity against Ad-36 were evaluated. Insulin resistance (IR) was determined according to criteria for the Chilean child population. Seropositivity against Ad-36 was determined by ELISA. Results: There was a high prevalence of overweight/obesity in the study population. Seropositivity against Ad-36 was 5.4% in the total group, but no association with nutritional status was observed. No correlation was found between Ad-36 seropositivity and lipid profile parameters. Insulin and HOMA-RI were significantly lower in the Ad-36 (+) group (p<0.001), and no cases of RI were reported in the Ad-36 (+) group in our population. Conclusions: Our results suggest that previous adenovirus-36 infection affects insulin secretion and insulin resistance, as previously described, however, no correlation is observed with the development of childhood obesity in the pediatric population. from southern Chile.

Management of acute respiratory distress syndrome in a child with adenovirus pneumonia: case report and literature review / Manejo da síndrome do desconforto respiratório agudo em criança com pneumonia por adenovírus: relato de caso e revisão da literatura

ABSTRACT Objective: To report the case of a child who developed acute respiratory distress syndrome (ARDS) from a pulmonary infection by adenovirus. Case description: A female patient aged 2 years and 6 months, weighting 10,295 grams developed fever, productive cough and vomiting, later on progressing to ARDS despite initial therapy in accordance with the institutional protocol for ARDS treatment. The child evolved to refractory hypoxemia and hypercapnia, requiring high parameters of mechanical pulmonary ventilation and use of vasoactive agents. In the treatment escalation, the patient received steroids, inhaled nitric oxide (iNO), was submitted to the prone position, started oscillatory high-frequency ventilation (HFOV) and extracorporeal membrane oxygenation (ECMO) was indicated due to severe refractory hypoxemia. During this time, the patient's clinical response was favorable to HFOV, improving oxygenation index and hypercapnia, allowing the reduction of vasoactive medications and mechanical ventilation parameters, and then the indication of ECMO was suspended. The patient was discharged after 26 days of hospital stay without respiratory or neurological sequelae. Comments: Adenovirus infections occur mainly in infants and children under 5 years of age and represent 2 to 5% of respiratory diseases among pediatric patients. Although most children with adenovirus develop a mild upper respiratory tract disease, more severe cases can occur. ARDS is a serious pulmonary inflammatory process with alveolar damage and hypoxemic respiratory failure; Adenovirus pneumonia in children may manifest as severe pulmonary morbidity and respiratory failure that may require prolonged mechanical ventilation. Exclusive pulmonary recruitment and HFOV are advantageous therapeutic options.

RESUMO Objetivo: Descrever paciente que evoluiu com síndrome do desconforto respiratório agudo (SDRA) a partir de infecção pulmonar por adenovírus. Descrição do caso: Paciente de dois anos e seis meses, sexo feminino, peso de 10295 g, que apresentou com quadro de febre, tosse produtiva e vômitos, evoluindo para SDRA. Apesar da terapêutica inicial em conformidade com o protocolo institucional de tratamento da SDRA, a criança evoluiu para hipoxemia e hipercapnia refratárias, necessitando de elevados parâmetros de ventilação pulmonar mecânica e utilização de agentes vasoativos. No escalonamento da terapêutica, a paciente recebeu terapias adjuvantes, foi iniciada ventilação oscilatória de alta frequência (VOAF) e indicada oxigenação por membrana extracorpórea (OMEC) pela hipoxemia grave refratária. Nesse ínterim, a paciente apresentou resposta clínica favorável à VOAF, melhorando do quadro ventilatório e possibilitando a redução das medicações vasoativas e dos parâmetros de ventilação mecânica. A paciente recebeu alta hospitalar após 26 dias de internação, sem sequelas respiratórias ou neurológicas. Comentários: As infecções por adenovírus ocorrem principalmente em lactentes e crianças com menos de cinco anos de idade e representam de 2 a 5% das doenças respiratórias entre os pacientes pediátricos. Embora a maioria das crianças com infecção por adenovírus desenvolva doença leve do trato respiratório superior, casos mais graves podem ocorrer com comprometimento do trato respiratório inferior. A pneumonia por adenovírus em crianças pode se manifestar com morbidade pulmonar grave e insuficiência respiratória com risco de vida, o que resulta na necessidade de suporte mecânico prolongado. O recrutamento pulmonar exclusivo pela VOAF pode ser uma opção terapêutica útil.

Fatal nosocomial hemorrhagic enterocolitis probably caused by adenovirus: report of one case / Enterocolitis hemorrágica fatal nosocomial probablemente causada por adenovirus: informe de un caso

ABSTRACT Adenovirus (ADV) is a recognized cause of severe disease among immunocompromised patients. We report a previously healthy 39-year-old female, admitted with influenza pneumonia and evolving with lung hemorrhage and acute renal failure requiring mechanical ventilation and hemodialysis. She received high corticosteroid doses due to an initial suspicion of alveolar hemorrhage. Lymphopenia already present before steroid use (567/μL), was maintained during the whole hospital stay (mean 782/μL). From the second week of admission she presented a high-volume diarrhea (mean 2.5 L/day) associated to intermittent bloody stools. An ulcerative enterocolitis was confirmed by CT images and colonoscopy. ADV was detected in a colonic tissue sample by real time PCR but not by a commercial filmarray test. Cidofovir-probenecid and racecadotril therapy were indicated without changing the clinical course of diarrhea and the patient finally died.

Adenovirus (ADV) es una causa reconocida de enfermedades graves en pacientes inmunocomprometidos. Informamos el caso de una mujer de 39 años, previamente sana, que ingresó por neumonía grave por influenza, evolucionando con hemorragia pulmonar y falla renal aguda, requiriendo ventilación mecánica y hemodiálisis. Recibió altas dosis de corticoides por la sospecha inicial de una hemorragia alveolar. Tuvo linfopenia durante toda su estadía (promedio 782/μL), la que ya estaba presente antes del uso de los corticoides (567/μL). Desde la segunda semana de hospitalización, presentó una diarrea de alto volumen (promedio 2,5 L/día) asociada a la presencia de sangre en deposiciones en forma intermitente. Se confirmó una enterocolitis ulcerativa por tomografía computada y colonoscopía. Se detectó ADV en muestras de biopsia colónica por PCR en tiempo real pero no por un test de PCR múltiples automatizado comercial. Fue tratada con cidofovir-probenecid y racecadrotrilo sin impacto clínico y la paciente finalmente falleció.

Molecular identification of viral agents associated with acute conjunctivitis: a prospective controlled study

Abstract Background: Viral conjunctivitis are the most frequent infections in ophthalmology clinics. The diagnosis is usually relying on clinical findings and medical history. However, topical antibiotics are often used unnecessarily addition to symptomatic treatment because of unsure agents. We aimed to detect the Adenovirus, Coxsackievirus and Enterovirus from conjunctiva and pharyngeal samples of patients. Methods: The conjunctiva and pharyngeal samples of the patients with conjunctivitis were taken by Virocult transport media and kept at -80 ºC up to study day. Adenovirus spp, Enterovirus 70 and Enterovirus 71, Coxsackie A24 and Coxsackie A16 were detected by real-time PCR. Samples from healthy health care workers of ophthalmology clinic were used for control group. Results: A total of 176 samples (conjunctival and pharyngeal samples of 62 patient and 26 healthy subjects) were included. The mean age of 34 (55.7%) male and 27 (44.3%) female patients was 34 ± 17. Twenty five (40.3%) of the patients were receiving antibiotic drops at first visit. The main etiologic agent in conjunctival samples was found to be Adenovirus (46/62, 74.2%) followed by Enterovirus 70 (4/62, 6.4%) and Enterovirus 71 (4/62, 6.4%). Coxsackievirus 16 and 24 were also found in 2 patients (1/62 each, 1.6%). Pharyngeal samples were also positive for Adenovirus (20/62, 32.3%), Enterovirus 70 and 71 (7/62, 11.3% and 5/62, 8.1% respectively), Coxsackievirus 16 and 24 (2/62, 3.2% and 1/61, 1.6%). Conclusions: It is very difficult in viral conjunctivitis to make clinical differentiation caused by different agents because of common clinical signs and symptoms. In routine clinical work, the viral conjunctivitis usually related with Adenovirus. But almost one fourth of the patients' conjunctivitis were not related to Adenovirus, which shows the importance of the laboratory diagnostics. True diagnosis plays an important role on prevention of contamination and unnecessary use of antibiotics in viral conjunctivitis.

A non-enteric adenovirus A12 gastroenteritis outbreak in Rio de Janeiro, Brazil

A gastroenteritis outbreak that occurred in 2013 in a low-income community in Rio de Janeiro was investigated for the presence of enteric viruses, including species A rotavirus (RVA), norovirus (NoV), astrovirus (HAstV), bocavirus (HBoV), aichivirus (AiV), and adenovirus (HAdV). Five of nine stool samples (83%) from patients were positive for HAdV, and no other enteric viruses were detected. Polymerase chain reaction products were sequenced and subjected to phylogenetic analysis, which revealed four strains and one strain of non-enteric HAdV-A12 and HAdV-F41, respectively. The HAdV-A12 nucleotide sequences shared 100% nucleotide similarity. Viral load was assessed using a TaqMan real-time PCR assay. Stool samples that were positive for HAdV-A12 had high viral loads (mean 1.9 X 107 DNA copies/g stool). All four patients with HAdV-A12 were < 25 months of age and had symptoms of fever and diarrhoea. Evaluation of enteric virus outbreaks allows the characterisation of novel or unique diarrhoea-associated viruses in regions where RVA vaccination is routinely performed.

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.

Adenoviruses of canine and human origins in stool samples from free-living pampas foxes (Lycalopex gymnocercus) and crab-eating foxes (Cerdocyon thous) in São Francisco de Paula, Rio dos Sinos basin / Adenovírus de origens canina e humana em fezes de graxains (Lycalopex gymnocercus) e cachorros-do-mato (Cerdocyon thous) de vida livre em São Francisco de Paula, bacia do Rio dos Sinos

The spread of enteric viruses of domestic animals and human beings to wild species can be facilitated by the resistance of these viruses on the environment and their ability to be transmitted by water and contaminated food. The health status of the populations of pampas foxes (Lycalopex gymnocercus) and crab-eating foxes (Cerdocyon thous) is largely unknown and the landscapes occupied by these animals in southern Brazil have been threatened by human occupation and expansion of agriculture. In this work, the search of genomes of human and canine adenoviruses in feces from these wild carnivores was used to track the dissemination of domestic animals and human pathogens to the free-living populations in a wildlife reserve located in southern Brazil. This was performed by virus-specific differential real-time polymerase chain reactions (qPCR) on stool specimens, avoiding capture and additional stress to the animals. Genus-specific conventional reverse-transcriptase PCR (RT-PCR) was complementarily performed aiming the detection of enteroviruses (EV) and rotaviruses (RV) on these same samples. HAdV genomes were found on 14 out of the 17 (82.35%) stool samples analysed, whereas CAV was found co-infecting 5 of these samples. RV genomes were detected on 7 of the 17 samples (41.18%) and all samples were negative for EV. The results point to the dispersion of HAdV and RV at a high rate to these species of South American wild carnivores, which can be an effect of growing anthropisation of the habitat of these animals.

A disseminação de vírus entéricos de animais domésticos e seres humanos para espécies selvagens pode ser facilitada pela resistência desses vírus no ambiente e sua capacidade de ser transmitida por água e alimentos contaminados. O estado de saúde das populações de Graxains-do-campo (Lycalopex gymnocercus) e Cachorros-do-mato (Cerdocyon thous) é em grande parte desconhecida e as paisagens ocupadas por estes animais no sul do Brasil têm sido ameaçadas pela ocupação humana e a expansão da agricultura. Neste trabalho, utilizou-se a pesquisa de genomas de adenovírus humanos (HAdV ) e caninos (CAV-1 e -2) em amostras fezes desses carnívoros selvagens com vistas a diagnosticar a disseminação de patógenos de animais domésticos e seres humanos às populações de vida livre em uma reserva de vida selvagem, localizado no sul do Brasil. Foram realizadas reações em cadeia da polimerase diferenciais e em tempo real (qPCR) de adenovírus específicos em amostras de fezes, evitando a captura e estresse adicional para os animais. PCRs gênero-específicas convencionais com transcrição reversa prévia (RT-PCR) foram ainda realizadas visando a detecção de enterovírus (EV) e rotavírus (RV) nestas mesmas amostras. Genomas de HAdV foram encontrados em 14 a 17 amostras de fezes (82.35%) analisados, Considerando que o CAV foi encontrado coinfectando 5 destas amostras. Genomas de RV foram detectados em 7 das 17 amostras (41.18%) e todas as amostras foram negativas para EV. Os resultados apontam para a dispersão de HAdV e RV em uma taxa elevada para estas espécies de carnívoros selvagens sul-americanas, que podem ser um efeito da crescente antropização do habitat desses animais.

Identificación de virus respiratorios en pacientes pediátricos / Identification of respiratory viruses in pediatric patients

Los virus son la principal causa de infecciones respiratorias agudas. Para la detección de los virus respiratorios se emplea la Inmunofluorescencia directa (IFD). Objetivos: Describir los resultados de las IFD positivas según edad de los pacientes y estacionalidad y determinar el porcentaje de positividad. Material y métodos: Estudio descriptivo retrospectivo. Se incluyeron todas las IFD realizadas en pacientes <15 años en el Hospital de Niños de Viña del Mar entre enero 2012 y noviembre 2013. Resultados: Se realizaron 32015 IFD en 9012 pacientes en ambos años. El virus más frecuente es el Virus Respiratorio Sincicial (VRS) (51 por ciento 2012, 63 porciento 2013), obteniendo un mayor número de casos en pacientes menores de 1año. El Adenovirus (ADV) ocupa el segundo lugar con una incidencia uniforme a lo largo del año. Se observó una disminución de los casos de Influenza A y B desde el año 2012 al 2013. El porcentaje de positividad global de los exámenes es muy bajo (4,25 por ciento 2012 y 3,19 por ciento 2013). El VRS alcanza la mayor positividad en invierno (31,8 por ciento 2012 y24,7 por ciento 2013) y llega al 0 por ciento durante el verano. Conclusiones: Nuestros resultados se asemejan a la epidemiología nacional principalmente en la distribución de los virus según edad y estacionalidad. El virus más frecuentemente encontrado es el VRS. Hubo menos casos de Influenza A durante el 2013 en comparación con los resultados en el país. Existe una solicitud aumentada de exámenes lo que lleva a un porcentaje de positividad muy bajo...

The main cause of acute respiratory infections is viral. The direct immunofluorescence (IF) is used in the detection of respiratory viruses. Objectives: To describe the results of positive IF in relationship to the patient´s age and seasonality and to determine the percentage of positive results. Material and Methods: A descriptive retrospective study that includes all IF performed in patients <15 years old in a private clinic in the V region from January 2012 to November 2013. Results: 32015 IF were conducted in 9012 patients during both years. The Respiratory Syncytial Virus (RSV) is the most common virus (51 percent 2012, 63 percent 2013), obtaining a larger number of cases in patients under the age of 1. The second most common virus is Adenovirus (ADV) with a uniform incidence during the year. A decrease in cases of Influenza A and B was observed from 2012 to 2013. The percent positivity of the tests is very low (4.25 percent and 3.19 percent 2012 2013). The RSV positive testing is higher during the winter (31.8 percent and 24.7 percent 2012 2013) and 0 percent during the summer. Conclusions: Our results are similar to the ones obtained by the national epidemiology in terms of age and season relationship. The RSV is the most common virus. There were fewer cases of influenza A in 2013 compared with results in the country. Due to high demand in IF testing, a very low percentage of positive results is observed...

Detección de norovirus en niños con diarrea adquirida en la comunidad o nosocomial en el Hospital Guillermo Grant Benavente de Concepción, Chile / Norovirus detection in children with community-acquired or nosocomial diarrhea at Guillermo Grant Benavente Hospital in Concepción, Chile

Introduction: Norovirus (NoV) are RNA viruses highly contagious, stable in the environment, genetically variable, and the most common cause of viral sporadic acute gastroenteritis worldwide. This is the first study carried out in Concepcion, Chile, to investigate the presence of NoV as an etiologic agent of viral diarrheas in hospitalized children. Objective. To detect the presence and genogroup of NoV in children with diarrhea and to compare it with rotavirus (RV) and adenovirus (AdV). Material and Methods: A one year descriptive, prospective study in children 0-14 years old. A single diarrheic stool sample per patient was analyzed for the presence of NoV, RV and AdV. Clinical data were unknown at the moment of sampling. Real time RT-PCR with Taqman™ probes for NoV and the immunocromatography VIKIA™ kit for RoV /AV detection were used. Results: Infection for NoV (25.5%) was significantly higher than for RV (15.9%) and AdV (6.2%). It was even greater in infants younger than 2yr. old (n: 103): NoV 34%, RV 17.5%, AdV 7.8%. Children 2-4 yr. old had 11.8% infection of NoV and RV. Children older than 4, only had 12% RV and 4% AdV. Children hospitalized for diarrhea (n: 92) had: 21.7% of both NoV and RV, and 7.6% AdV; whereas children hospitalized for other causes (n: 53) had 32.1% NoV,5.7% RV and 3.8% AV. The proportion of infection due to NoV was significantly higher in males (31.5%) than in females (19.4%). The average frequency during the year was higher for NoV (30.3%) than for RV (14.7%) except in summer. Conclusion: The presence of NoV was higher than RV in children with diarrhea. NoV infection showed defined characteristics regarding age, gender, seasonal occurrence and nosocomial transmission that are important epidemiological features.

Introducción: Los norovirus (NoV) son virus ARN altamente contagiosos, resistentes, variables genéticamente y una de las etiologías más frecuente de gastroenteritis viral esporádica mundial. Este es el primer trabajo en Concepción, Chile, de búsqueda de NoV como etiología viral de diarreas en niños hospitalizados. Objetivo: Determinar la presencia y genogrupo de NoV en niños con diarrea y compararla con la frecuencia de rotavirus (RV) y adenovirus (AdV). Material y Método: Estudio descriptivo, prospectivo de un año, en niños de 0-14 años ingresados por diarrea aguda o que la adquirieron dentro del hospital. La muestra de deposiciones diarreica se tomó una sola vez por paciente. Las fichas clínicas se analizaron al finalizar el estudio etiológico. Para la detección de NoV se utilizó RPC-TR a en tiempo real con sondas Taqman® y para detección de RV/AdV, el kit VIKIA® de inmunocromatografia. Resultados: La infección por NoV (25,5%) fue significativamente más frecuente que por RV (15,9%) y AdV (6,2%). La mayor presencia de infección fue en pacientes bajo2 años de edad (n: 103): NoV 34,0%, RV 17,5%, AdV 7,8%. La detección en niños hospitalizados por diarrea fue: NoV y RV 21,7% cada uno; AdV 7,6%. En niños con diarrea nosocomial hospitalizados por otras causas se detectó NoV en 32,1%, RV en 5,7% y AdV en 3,8%. La presencia de NoV fue significativamente mayor en varones (31,5%) que en niñas (19,4%). El promedio de diarreas durante el año fue mayor para NoV (30,3%) que para RV(14,7%), excepto en verano. Discusión y Conclusión: La presencia de NoV fue mayor que la de RoV en niños con diarrea y con una tendencia nosocomial que podría deberse a las características del virus que favorece infecciones de ambiente confinado, como hospitales, asilos y cruceros. La infección por NoV presentó características definidas, en edad, género, ocurrencia estacional y relevancia nosocomial, que aportan datos epidemiológicos importantes.

Epidemiological aspects of influenza A related to climatic conditions during and after a pandemic period in the city of Salvador, northeastern Brazil

During the influenza pandemic of 2009, the A(H1N1)pdm09, A/H3N2 seasonal and influenza B viruses were observed to be co-circulating with other respiratory viruses. To observe the epidemiological pattern of the influenza virus between May 2009-August 2011, 467 nasopharyngeal aspirates were collected from children less than five years of age in the city of Salvador. In addition, data on weather conditions were obtained. Indirect immunofluorescence, real-time transcription reverse polymerase chain reaction (RT-PCR), and sequencing assays were performed for influenza virus detection. Of all 467 samples, 34 (7%) specimens were positive for influenza A and of these, viral characterisation identified Flu A/H3N2 in 25/34 (74%) and A(H1N1)pdm09 in 9/34 (26%). Influenza B accounted for a small proportion (0.8%) and the other respiratory viruses for 27.2% (127/467). No deaths were registered and no pattern of seasonality or expected climatic conditions could be established. These observations are important for predicting the evolution of epidemics and in implementing future anti-pandemic measures.

A simple negative selection method to identify adenovirus recombinants using colony PCR

Background: The AdEasy system is a fast-track system for generating recombinant adenoviruses using the efficient homologous recombination machinery between shuttle and adenovirus backbone plasmids in Escherichia coli BJ5183 cells. The key step is homologous recombination in BJ5183 cells, which is driven by RecA activity. However, culture time is stringently limited to reduce the damage to recombinant plasmids by RecA activity. Therefore, rapid identification of recombinant adenoviruses within the limited time-period is critical. Results: We developed a simple negative selection method to identify recombinant adenoviruses using colony PCR, which improves the efficiency of adenovirus recombination screening and packaging. Conclusions: The negative selection method to identify AdEasy adenovirus recombinants by colony PCR can identify the recombined colony within a short time-period, and maximally avoid damage to the recombinant plasmid by limiting recombination time, resulting in improved adenovirus packaging.

The efficiency of concentration methods used to detect enteric viruses in anaerobically digested sludge

The presence of enteric viruses in biosolids can be underestimated due to the inefficient methods (mainly molecular methods) used to recover the viruses from these matrices. Therefore, the goal of this study was to evaluate the different methods used to recover adenoviruses (AdV), rotavirus species A (RVA), norovirus genogroup II (NoV GII) and the hepatitis A virus (HAV) from biosolid samples at a large urban wastewater treatment plant in Brazil after they had been treated by mesophilic anaerobic digestion. Quantitative polymerase chain reaction (PCR) was used for spiking experiments to compare the detection limits of feasible methods, such as beef extract elution and ultracentrifugation. Tests were performed to detect the inhibition levels and the bacteriophage PP7 was used as an internal control. The results showed that the inhibitors affected the efficiency of the PCR reaction and that beef extract elution is a suitable method for detecting enteric viruses, mainly AdV from biosolid samples. All of the viral groups were detected in the biosolid samples: AdV (90%), RVA, NoV GII (45%) and HAV (18%), indicating the viruses' resistance to the anaerobic treatment process. This is the first study in Brazil to detect the presence of RVA, AdV, NoV GII and HAV in anaerobically digested sludge, highlighting the importance of adequate waste management.

Evaluation of a rapid immunochromatographic assay for the detection of rotavirus, norovirus and adenovirus from children hospitalized with acute watery diarrhoea

We evaluated the IP-Triple I immunochromatographic rapid test for the detection of rotavirus, norovirus and adenovirus using stool samples from children with diarrhoea. The detection of norovirus and adenovirus was poor compared to polymerase chain reaction assays. However, high sensitivity (92%) and specificity (99%) were obtained for the detection of rotavirus.

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.

Detection of enteric viruses in recreational waters of an urban lagoon in the city of Rio de Janeiro, Brazil

The aim of this study was to determine the occurrences of the group A rotavirus (RVA), norovirus (NoV) and human adenovirus (HAdV) in the surface waters of an urban lagoon (Rodrigo de Freitas Lagoon) in the city of Rio de Janeiro, Brazil. During one year of surveillance, water samples were obtained from the lagoon and other interconnected ecosystems (river and beach). The samples were concentrated using an adsorption-elution method with a negatively charged membrane and tested by qualitative and quantitative polymerase chain reaction assays. RVA was the most prevalent virus detected (24.3%) with a viral load ranging from 3.0 x 10¹-5.6 x 10(4) genome copies/L, followed by NoV (18.8%) and HAdV (16.7%). Considering water samples suitable for bathing, according to Escherichia coli criterion (< 2,000 most probable number/100 mL), viruses were detected in 50% (57/114) of them. Physicochemical parameters were also measured and showed possible correlations between turbidity and RVA presence and between pH and NoV presence. These data demonstrate the importance of considering viral parameters to ensure water quality and the utilisation of these parameters as additional tools for the characterisation of environmental contamination.

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.

Research of Viral Agent in Free-living Pigeon Feces (Columba livia) in the City of São Paulo, SP, Brazil, for Transmission Electron Microscopy / Investigación de Agentes Virales en Heces de Palomas de Vida Libre (Columba livia) en la Ciudad de São Paulo, SP, Brasil, Através de Miscroscopía Electrónica de Transmisión

The pigeon feces are vehicle of diseases both for humans and other animal species. In these birds, the most important viral diseases of the digestive tract are transmitted by the paramyxovirus, adenovirus and coronavirus. Avian paramyxoviruses have been isolated from a variety of species of free living and domestic birds worldwide, with several symptoms and clinical signs and economic losses. Paramyxoviruses belong to the Paramyxoviridae family and Avulovirus genus that includes nine serotypes (APMV 1 to 9). Avian adenoviruses belong to the Adenoviridae family and Aviadenovirus genus. In pigeons, cause classical adenovirosis and necrotizing hepatitis. The respiratory and enteric tracts are common targets of coronavirus. They belong to the Coronaviridae family and to 3a and 3c groups. In this study, we described the presence of viral agents in free-living pigeon feces (Columba livia) from the city of São Paulo, Brazil. The feces were processed by negative staining technique (rapid preparation) for transmission electron microscopy. In this technique paramyxoviruses particles, pleomorphic, roughly spherical or filamentous, measuring 100 to 500 nm of diameter containing an envelope covered by spikes, with characteristic helical herring-bone-like nucleocapsid, measuring 15 to 20 nm in diameter, were visualized in 45 (79 percent) out of 57 feces samples. In 2 (3.5 percent) samples, paramyxovirus and adenovirus particles were simultaneously visualized. Adenovirus particles were isometric, spherical, characterized as "complete "or" empty ", measuring between 70 and 90 nm in diameter. Paramyxovirus and coronavirus particles were detected in 3 (5.2 percent) samples. Coronaviruses were pleomorphic with a diameter of 75-160 nm containing a solar corona-shaped envelope, with projections of approximately 20 nm of diameter. Seven (12.3 percent) samples were negative for viral particles.

Los heces de las palomas constituyen vehículos de enfermedades importantes, tanto para el Hombre como para otras especies animales. En estas aves, las enfermedades virales más importantes del tracto digestivo son transmitidas por los paramixovirus, adenovirus y coronavirus. Paramixovirus aviario, en todo el mundo, ha sido aislado de una variedad de especies de vida libre y de aves domésticas, que causan variados síntomas y señales clínicas con pérdidas económicas. El Paramixovirus pertenece a la familia Paramyxoviridae y al género Avulavirus que incluye nueve serotipos (APMV 1 a 9). Adenovirus aviario pertenece al género de la familia Adenoviridae y género Aviadenovirus. En las palomas, causan la adenovirosis clásica y la hepatitis necrotizante. El tracto respiratorio y entérico son los albos comunes de los coronavirus. Ellos pertenecen a la familia Coronaviridae y a los grupos 3a y 3c. En este trabajo, se describe la presencia de agentes virales en las heces de palomas de vida libre (Columba livia) en la ciudad de São Paulo, Brasil. Las heces se procesaron para la técnica de microscopía electrónica de transmisión, a través de la técnica de contrastación negativa (preparación rápida). A través de esta técnica fueron visualizadas las partículas de paramixovirus, pleomórficas, más o menos esféricas o filamentosas, de 100 a 500 nm de diámetro que contiene un envoltorio cubierto con espículas, con nucleocapside con características helicoidales, midiendo de 15 a 20 nm de diámetro en 45 (79 por ciento) de 57 muestras. En 2 (3,5 por ciento) muestras, fueron observadas simultáneamente partículas de paramixovirus y de adenovirus. Las partículas de adenovirus eran isométricas, esféricas, caracterizadas como “completa " o “vacía ", midiendo entre 70 y 90 nm de diámetro. Fueron analizadas en tres muestras (5,2 por ciento) las partículas de paramixovirus y coronavirus. Los coronavirus son pleomórficos, con un diámetro de 75 a 160 nm, que contiene un capa en forma de corona solar con proyecciones de aproximadamente 20 nm de diámetro. Siete (12,3 por ciento) muestras fueron negativas para las partículas virales.

Gastroenteric virus detection in fecal samples from women in Goiânia, State of Goiás, Brazil / Detecção de vírus gastroentéricos em amostras fecais de mulheres em Goiânia, Estado de Goiás, Brasil

INTRODUCTION: This was a prospective study that included women seen in the obstetrics and gynecology sector of Hospital das Clínicas, Federal University of Goiás, in Goiânia, State of Goiás, with the aim of detecting rotaviruses, adenoviruses, caliciviruses and astroviruses. Eighty-four women participated in the study and from these, 314 fecal samples were collected. Out of all of the women, 29 were seropositive for HIV and 55 were seronegative, and 45 and 39 were pregnant and non-pregnant, respectively. METHODS: Fecal samples were collected from each woman once every two months over the period from July 2006 to June 2007, and they were screened for rotaviruses by means of polyacrylamide gel electrophoresis and immunoenzymatic assays, for caliciviruses and astroviruses by means of RT-PCR and for adenovirus by means of immunoenzymatic assays. The astroviruses were genotyped using nested PCR. RESULTS: Among the 84 patients, 19 (22.6 percent) were positive for either calicivirus (14/19) or astrovirus (6/19), while one women was positive for both viruses in fecal samples collected on different occasions. Most of the positive samples were collected during the months of July and August (astrovirus) and September and October (calicivirus). None of the samples analyzed was positive for rotavirus or adenovirus. Gastroenteric viruses were detected in 13/19 (68.4 percent) of the pregnant women, whether HIV-seropositive or not. CONCLUSIONS: The results from the present study showed that neither pregnancy nor HIV-seropositive status among the women increased the risk of infection by any of the gastroenteric viruses studied. This study presents data on gastroenteric virus detection among pregnant and/or HIV-positive women.

INTRODUÇÃO: Este foi um estudo prospectivo que incluiu mulheres atendidas no setor de obstetrícia e ginecologia do Hospital das Clínicas da Universidade Federal de Goiás, em Goiânia, Estado de Goiás com o objetivo de detectar rotavírus, adenovírus, calicivírus e astrovírus. Oitenta e quatro mulheres participaram no estudo e destas, 314 amostras fecais foram coletadas. Do total de mulheres, 29 eram soropositivas para HIV, 55 soronegativas, 45 e 39 estavam grávidas e não-grávidas, respectivamente. MÉTODOS: Amostras fecais foram coletadas de cada mulher uma vez a cada dois meses pelo período de Julho-2006 a Junho-2007, foram triadas para rotavírus pela metodologia de eletroforese em gel de poliacrilamida (EGPA) e através de ensaio imunoenzimático (EIE), para calicivírus e astrovírus por RT-PCR e por EIE para adenovírus. Os astrovírus foram genotipados por Nested-PCR. RESULTADOS: De 84 pacientes, 19 (22,6 por cento) foram positivas para calicivírus (14/19) ou astrovírus (6/19), sendo que uma mulher foi positiva para ambos os vírus em amostras fecais coletadas em diferentes ocasiões. A maioria das amostras positivas foi coletada no período de Julho a Agosto (astrovírus) e de Setembro a Outubro (calicivírus). Nenhuma das amostras analisadas foi positiva para rotavírus ou adenovírus. Os vírus gastroentéricos foram detectados em 13/19 (68,4 por cento) mulheres grávidas, as quais eram HIV-soropositivas ou não. CONCLUSÕES: Os resultados do presente estudo mostram que nem o estado gravídico das mulheres nem a soropositividade para HIV aumentaram o risco para a infecção por nenhum dos vírus gastroentéricos estudados. Este estudo apresenta dados sobre a detecção de vírus gastroentéricos entre mulheres grávidas e/ou HIV-positivas.