Identification of enteric viruses circulating in a dog population with low vaccine coverage

ABSTRACT Although the use of vaccines has controlled enteric diseases in dogs in many developed countries, vaccine coverage is still under optimal situation in Brazil. There is a large population of nonimmunized dogs and few studies about the identification of the viruses associated with diarrhea. To address this situation, stool samples from 325 dogs were analyzed by polymerase chain reaction for the detection of common enteric viruses such as Canine adenovirus (CAdV), Canine coronavirus (CCoV), Canine distemper virus (CDV), Canine rotavirus (CRV) and Carnivorous protoparvovirus 1 (canine parvovirus 2; CPV-2). At least one of these species was detected in 56.6% (184/325) of the samples. The viruses detected most frequently in either diarrheic or nondiarrheic dog feces were CPV-2 (54.3% of the positive samples), CDV (45.1%) and CCoV (30.4%), followed by CRV (8.2%) and CAdV (4.9%). Only one agent was detected in the majority of the positive samples (63%), but co-infections were present in 37% of the positive samples and mainly included CDV and CPV-2. The data presented herein can improve the clinical knowledge in regions with low vaccine coverage and highlight the need to improve the methods used to control these infectious diseases in domestic dogs.

Virus más frecuentemente aislados en la exacerbación de asma y su correlación con nivelesde eosinófilos e inmunoglobulina E sérica total / Most commonly isolated viruses in asthma exacerbation and their correlation with eosinophil and total serum immunoglobulin E levels

Antecedentes. Las exacerbaciones de asma continúan siendo una causa de hospitalización en el Servicio de Urgencias. Los desencadenantesson alérgenos e infecciones, principalmente, de tipo viral. El objetivo fue determinar la relación entre los virus detectados durante la exacerbación asmática y los niveles de eosinófilos e inmunoglobulina E (IgE) sérica en pacientes pediátricos. Población y métodos. Estudio transversal analítico. Se incluyeron niños de cinco a quince años atendidos en Urgencias de Pediatría con exacerbación de asma, en el período de marzo de 2013 a febrero de 2016. Se obtuvo ácido ribonucleico viral en el aspirado nasofaríngeo con el kit CLART PneumoVir. Se cuantificaron los eosinófilos en la sangre periférica y los niveles de IgE sérica total. Se consideró eosinofilia un conteo ≥ 0,4 x 103/mm3 e IgE elevada, ≥ 350 UI/L. Se realizó la correlación de Pearson. Se definió significancia con valor de p ≤ 0,05.Resultados. De 211 niños con exacerbación de asma, en el 20%, se aisló un virus. Los virus aislados más frecuentemente fueron el rinovirus, el enterovirus y el virus sincitial respiratorio. Se encontró una correlación entre los niveles de eosinófilos e IgE sérica total en los niños con exacerbación de asma y rinovirus de 0,89, con una p= 0,0001.Conclusiones. Las infecciones por rinovirus, enterovirus y virus sincitial respiratorio son más frecuentes en las exacerbaciones de asma en menores de 15 años. Se observó una correlación entre los niveles de eosinófilos e IgE en presencia de rinovirus.

Background. Asthma exacerbations are still a cause of hospitalization at the Emergency Department. The triggers of asthma exacerbations include allergens and infections ­mostly viral­. The objective of this study was to establish the relationship between viruses detected during an asthma exacerbation and eosinophil and serum immunoglobulin E (IgE) levels in pediatric patients. Population and methods. Cross-sectional. analytical study. Children aged 5-15 years seen at the Pediatric Emergency Department with an asthma exacerbation in the period between March 2013 and February 2016 were included. Viral ribonucleic acid was extracted from nasopharyngeal aspirates using the CLART Pneumo Vir kit. Eosinophil levels were measured in peripheral blood and total IgE levels, in serum. Eosinophilia was defined as a count ≥ 0.4 x 103/mm3 and high IgE. as a level ≥ 350 IU/L. The Pearson's correlation was carried out. A value of p ≤ 0.05 was considered significant.Results. Out of 211 children with asthma exacerbation, a virus was isolated in 20%. The most commonly isolated viruses were rhinovirus. enterovirus, and respiratory syncytial virus. A correlation of 0.89 was established between eosinophil and total serum IgE levels in children with asthma exacerbation and rhinovirus, with a p value of 0.0001. Conclusions. Rhinovirus, enterovirus, and respiratory syncytial virus were the most common viruses in asthma exacerbations in children younger than 15 years. A correlation was established between eosinophil and IgE levels in the presence of rhinovirus.

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.

Comparison of enterovirus detection in cerebrospinal fluid with Bacterial Meningitis Score in children / Comparação da pesquisa de enterovírus no liquor com Escore de Meningite Bacteriana em crianças

ABSTRACT Objective To measure the role of enterovirus detection in cerebrospinal fluid compared with the Bacterial Meningitis Score in children with meningitis. Methods A retrospective cohort based on analysis of medical records of pediatric patients diagnosed as meningitis, seen at a private and tertiary hospital in São Paulo, Brazil, between 2011 and 2014. Excluded were patients with critical illness, purpura, ventricular shunt or recent neurosurgery, immunosuppression, concomitant bacterial infection requiring parenteral antibiotic therapy, and those who received antibiotics 72 hours before lumbar puncture. Results The study included 503 patients. Sixty-four patients were excluded and 94 were not submitted to all tests for analysis. Of the remaining 345 patients, 7 were in the Bacterial Meningitis Group and 338 in the Aseptic Meningitis Group. There was no statistical difference between the groups. In the Bacterial Meningitis Score analysis, of the 338 patients with possible aseptic meningitis (negative cultures), 121 of them had one or more points in the Bacterial Meningitis Score, with sensitivity of 100%, specificity of 64.2%, and negative predictive value of 100%. Of the 121 patients with positive Bacterial Meningitis Score, 71% (86 patients) had a positive enterovirus detection in cerebrospinal fluid. Conclusion Enterovirus detection in cerebrospinal fluid was effective to differentiate bacterial from viral meningitis. When the test was analyzed together with the Bacterial Meningitis Score, specificity was higher when compared to Bacterial Meningitis Score alone.

RESUMO Objetivo Avaliar o papel da pesquisa de enterovírus no líquido cefalorraquidiano em comparação com o Escore de Meningite Bacteriana em crianças com meningite. Métodos Coorte retrospectiva, realizada pela análise de prontuários, incluindo pacientes pediátricos, com diagnóstico de meningite e atendidos em um hospital privado e terciário, localizado em São Paulo, entre 2011 e 2014. Foram excluídos os pacientes com doença crítica, púrpura, derivação ventricular ou neurocirurgia recente, imunossupressão, outra infecção bacteriana concomitante que necessitasse de antibioticoterapia parenteral e aqueles que receberam antibiótico 72 horas antes da punção lombar. Resultados Foram incluídos no estudo 503 pacientes. Destes, 64 foram excluídos e 94 não realizaram todos os exames para análise. Dos 345 pacientes restantes, 7 ficaram no Grupo de Meningite Bacteriana e 338 no Grupo de Meningite Asséptica. Não houve diferença estatística entre os grupos. Na análise do Escore de Meningite Bacteriana, dos 338 pacientes com possível meningite asséptica (culturas negativas), 121 deles tiveram um ou mais pontos para o Escore de Meningite Bacteriana, com valor de sensibilidade de 100%, especificidade de 64,2% e valor preditivo negativo de 100%. Dos 121 pacientes com Escore de Meningite Bacteriana positivo, 71% (86 pacientes) tiveram a pesquisa de enterovírus positiva no líquido cefalorraquidiano. Conclusão A pesquisa de enterovírus no líquido cefalorraquidiano mostrou-se eficaz em diferenciar a meningite bacteriana da viral. Analisada junto com o Escore de Meningite Bacteriana, a especificidade foi maior em comparação ao Escore de Meningite Bacteriana isolado.

Red blood cells in cerebrospinal fluid as possible inhibitory factor for enterovirus RT-PCR / Presença de hemácias no líquido cefalorraquidiano como possível fator inibitório da RT-PCR para detecção de enterovirus

ABSTRACT The presence of hemoglobin in samples are considered an important inhibitory factor for polymerase chain reaction (PCR). The aim of this study was to examine the influence of red blood cells (RBC)s in cerebrospinal fluid (CSF) as an inhibitory factor to reverse transcription polymerase chain reaction (RT-PCR) for enteroviruses (EV). Forty-four CSF samples from patients showing characteristics of viral meningitis were assessed for EV by RT-PCR. Viral RNA extracted with guanidine isothyocianate buffer and virus detection was performed by in-house nested PCR. Positivity for EV RT-PCR was higher in CSF samples without RBCs than in samples with RBCs: 13(26%) and 36(9.2%), p = 0.001. In the group with positive EV RT-PCR, the mean + SD CSF RBC was 37 ± 183 cell/mm3; the group with negative results had 580 + 2,890 cell/mm3 (p = 0.007). The acceptable upper limit for CSF RBCs that could not influence RT-PCR was 108 cells/mm3. CSF samples with negative results for EV RT-PCR have more erythrocytes.

RESUMO A presença de hemoglobina em amostras de fluidos corporais é considerada um fator inibitório importante da reação em cadeia da polimerase (PCR). O objetivo deste estudo era examinar a influencia de hemácias no líquido cefalorraquidiano (LCR) como um fator inibitório da RT-PCR para enterovirus (EV). Quatrocentos e quarenta amostras de LCR de pacientes com características de meningite viral foram avaliados para enterovirus por RT-PCR. RNA viral foi extraído com tampão de isotiocianato de guanidina e a detecção viral foi feita com nested PCR in-house. A positividade do EV RT-PCR no LCR foi maior nas amostras de LCR sem hemácias do que as amostras com hemácias: 13 (26%) e 36 (9,2%), respectivamente (p = 0,001). No grupo com resultados EV RT-PCR positivo, a media ± DP do número de hemácias no LCR foi 37 ± 183 cell/mm3 e no grupo com resultados negativos foi 580 ± 2.890 cell/mm3 (p = 0,007). O limite superior aceitável de hemácias no LCR para não inibir o resultado do PCR foi 108 cells/mm3. As amostras de LCR com resultados negativos para RT-PCR EV tem mais eritrócitos em comparação com amostras com resultados positivos.

Moving beyond classical markers of water quality: detection of enteric viruses and genotoxicity in water of the Sinos River / Indo além dos marcadores clássicos de qualidade da água: detecção de vírus entéricos e genotoxicidade na água do Rio dos Sinos

It is well recognized that the classical biological and chemical markers of environmental pollution do not necessarily indicate the presence or absence of emerging threats to public health, such as waterborne viruses and genotoxicants. The purpose of this preliminary study was to evaluate the presence of material of enteroviruses (EV), rotavirus (RV) and adenovirus (AdV) and genotoxicity in water samples from points of routine monitoring of water quality in the main course of the Sinos River. The points are classified into different levels of pollution in accordance to the Brazilian federal regulations. Viral genomes from EV, AdV were detected in two of the 4 collection points regardless of the level of urbanisation of the surrounding areas. In contrast, genotoxicity was not observed in piava (Leporinus obtusidens) fingerlings cultivated on these same water samples. Results were compared with classical physical, chemical and microbiological parameters. There was no clear evidence of association between any of the classical markers and the presence of viral genomes in the water samples tested.

É amplamente reconhecido que os marcadores biológicos e químicos clássicos para a poluição ambiental não necessariamente indicam a presença ou ausência de ameaças emergentes à saúde pública, tais como vírus transmitidos pela água e genotoxicantes. Este estudo preliminar teve por objetivo detectar material genético de enterovírus (EV), rotavírus (RV) e adenovírus (AdV) e genotoxicidade em amostras de água de pontos de monitoramento de rotina da qualidade da água no curso principal do rio dos Sinos. Os pontos são classificados em níveis diferentes de poluição, de acordo com as normativas federais brasileiras. Genomas virais de EV, RV e RV foram detectados em dois dos quatro pontos de coleta, independente do nível de urbanização das áreas adjacentes. Por outro lado, não foi observada genotoxicidade em alevinos de piava (Leporinus obtusidens) cultivados nestas mesmas amostras de água. Os resultados são comparados com marcadores físicos, químicos e microbiológicos clássicos, não há nenhuma evidência clara da associação entre qualquer um dos marcadores clássicos e da presença de genomas virais nas amostras de água testadas.

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.

La vacuna polio oral en lactantes no interfiere con la detección de enterovirus en sangre / Oral polio vaccine in infants does not interfere in detection of enterovirus in blood

Introduction: There is not known if a viraemia post-oral polio vaccine (OPV) is detectable by modern molecular techniques. Such viraemia could affect the performance of the real time-polymerase chain reaction (PCR) for non polio enterovirus (EV) detection, technique of growing clinical use for the study of febrile infants. Objective: To determine viraemia post-first dose of OPV in healthy infants, by molecular techniques. Patients and Methods: 50 infants less than three months without previous VPO were randomized in 5 groups: a control group with pre-vaccination blood sample (BS), group 1 BS at day 2, group 2 BS at day 4, group 3, BS at day 6 and group 4, BS at day 8 post-vaccination. Conventional and specific PCR for poliovirus and real time PCR for non polio EV were performed in BS and in OPV samples. Results: No genetic material of poliovirus was detected in any infant, while in 9 of them (18%) non polio EV was identified. Real time PCR for EV did not amplify poliovirus from OPV samples. Discussion: Results suggest that no post VPO viraemia detectable by molecular methods exists. Considering that real time PCR for EV does not allow to identify polio virus, no false positives of the test are expected as a result of a recent VPO vaccination. We documented presence of non polio EV in blood of healthy asymptomatic infants.

Introducción: No existen estudios que indiquen si la vacuna polio oral (VPO) produce viremia detectable mediante métodos moleculares. Una eventual viremia podría afectar el rendimiento de la RPC tiempo real para detectar enterovirus (EV) no polio, examen de creciente uso clínico en lactantes pequeños con fiebre sin foco. Objetivo: Determinar viremia post VPO en lactantes sanos, por métodos moleculares. Métodos: 50 menores de 3 meses, al momento de recibir su primera VPO se distribuyeron en forma aleatoria en 5 grupos: control, muestra de sangre pre-vacunación; grupo 1, muestra al 2° día; grupo 2, al 4° día; grupo 3, al 6° día y grupo 4, al 8° día post-vacunación. Se realizó RPC convencional específica para virus polio y RPC tiempo real para EV no polio en las muestras de sangre y en muestras de VPO. Resultados: No se identificó presencia de material genético de virus polio en lactante alguno, mientras que en 9 (18%) se identificó presencia de EV no polio. La RPC tiempo real para EV no polio no amplificó material genético a partir de las muestras de VPO. Discusión: Los resultados sugieren que no existe viremia post-VPO detectable por métodos moleculares. Considerando que la RPC tiempo real de EV no polio de uso clínico no permite identificar la presencia de virus polio, estos hallazgos indican que no existirán falsos positivos de este examen como resultado de una vacunación VPO reciente. Adicionalmente se documentó presencia de EV no polio en sangre de lactantes asintomáticos.

Detection of human adenovirus, rotavirus and enterovirus in water samples collected on dairy farms from Tenente Portela, Northwest of Rio Grande do Sul, Brazil

Viral gastroenteritis and other waterborne diseases are a major concern for health in Brazil. A number of studies were conducted about the presence of viruses on water samples from Brazilian areas. However, the knowledge about the occurrence of viral contamination of drinking water sources in rural settings of the country is insufficient. On the present work, 15 samples from 5 dairy farms located at the municipality of Tenente Portela were collected and analysed for the presence of human adenoviruses (HAdV), as well as human enteroviruses (EV) and rotaviruses (RV). HAdV was present on 66.66% of the water samples, and have been found in all samples from artesian wells and springs, which are used as sources of drinking water for the individuals inhabiting those farms. EV and RV found only in one sample each. The detection rates of HAdV on the water from these dairy farms are alarming and point towards a situation of elevated environmental contamination by fecal microorganisms of human origin and poor basic sanitation conditions.

Hand, foot, and mouth disease: Current scenario and Indian perspective.

Hand, foot, and mouth disease (HFMD), first reported in New Zealand in 1957 is caused by Coxsackievirus A16 (CVA16) and human enterovirus 71 (HEV71) and occasionally by Coxsackievirus A4-A7, A9, A10, B1-B3, and B5. This is characterized by erythematous papulo vesicular eruptions over hand, feet, perioral area, knees, buttocks and also intraorally mostly in the children. HFMD has been known for its self limiting course. Only small scale outbreaks have been reported from United States, Europe, Australia, Japan and Brazil for the first few decades. However, since 1997 the disease has conspicuously changed its behavior as noted in different Southeast Asian countries. There was sharp rise in incidence, severity, complications and even fatal outcomes that were almost unseen before that period. Following the near complete eradication of poliovirus, HEV71, the non-polio enterovirus, may become the greatest threat to cause significant neurological complications. This adds to the fact that effective therapy or vaccine is still a far reaching goal. There are reports of disease activity in different corners of India since 2004. Although of milder degree, continuous progress to affect larger parts of the country may indicate vulnerability of India from possible future fatal outbreaks. Low level of awareness among the health care providers may prove critical.

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 / 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.

Identificação de enterovírus humanos a partir de amostras fecais de crianças menores de 15 anos, atendidas no Hospital Geral de Mavalane na cidade de Maputo, Moçambique / Identification of human enteroviruses from stool samples from children under 15 years, the Hospital General Mavalane in Maputo, Mozambique

Os enterovírus humanos (HEV) são espécies do gênero Enterovirus,família Picornaviridae. Existem cerca de 120 sorotipos de HEV que são divididos em quatro espécies, designadas de HEV-A a D. Estes agentes infectam anualmente, milhões de pessoas no mundo, resultando em uma grande variedade de quadros clínicos que vão desde infecções inaparentes à febres inespecíficas, resfriado comum, à doenças graves, tais como meningite e poliomielite paralítica. As crianças são mais susceptiveis à infecção. A transmissão ocorre tanto pela via entérica e por via respiratória. O vírus pode ser excretado nas fezes por várias semanas. Este estudo teve como objectivo isolar e identificar os sorotipos de HEVs circulantes, a partir de amostras de fezes de crianças menores de 15 anos de idade, com quadros compatíveis a infecção por esses agentes, no Hospital Geral de Mavalane na Cidade de Maputo, em Moçambique. Neste trabalho,foram utilizadas 178 amostras de fezes obtidas entre novembro de 2011 a fevereiro de 2012.As amostras foram inoculadas em culturas de células e os enterovírus isolados foram identificados através de metódos moleculares, nas amostras negativas foi pesquisado o adenovírus. Das 45 amostras positivas em cultivos celulares, os enterovírus foram isolados e identificados em 26 (14,6 (por cento)). A proporção sexo masculino e feminino foi de 1,8: 1. O isolamento dos enterovírus diminuiu à medida que a idade aumentou. O sequenciamento gênomico revelou uma grande diversidade de enterovírus humanos. Entre os 26 enterovírus isolados, o Echovírus 29 foi o agente mais identificado com 19,2 (por cento), seguido pelo Enterovírus 99 (11,5(por cento)). Foram identificados também Coxsackievírus A5, Echovírus sorotipos 11, 13 e Enterovírus C com 7,7 (por cento) de cada ; Coxsackievírus sorotipos A10, A13, A20, B4 eB6 com 3,85 (por cento) cada; Echovírus sorotipos 7, 21 e 25, com 3,85 (por cento) cada um, e Poliovírus sorotipos 2 e 3 com 3,85 (por cento), respectivamente. Adenovírus foram isolados em 20 amostras, representando 11,2 (por cento) do total (20/178). Duas amostras apresentaram co-infecção enterovírus/adenovírus. Os resultados deste trabalho evidenciam a circulação de uma grande diversidade enterovírus humanos na cidade deMaputo, sendo os echovírus mais frequentes, mas também mostra a circulação de adenovírus humanos. Outros testes laboratoriais seriam necessários, para se relacionar inequivocamente a participação desses agentes virais na etiologia dos quadros clínicos observados.

Detecção e quantificação de entrerovirus em lodo de esgoto proveniente de estações de tratamento de esgotos com potencial uso na agricultura do Estado de São Paulo / Dectetion of enterovirus in sewage sludge from sewage plant tratament with potential use in agriculture in São Paulo State

Lodos de esgoto são resíduos do tratamento do esgoto doméstico, considerados ricos em macronutrientes e matéria orgânica, podendo também apresentar contaminação por substâncias químicas e patógenos. Sua utilização na agricultura é uma das alternativas interessantes para sua disposição final. Entretanto, a presença de contaminantes, dentre eles microrganismos patogênicos podem limitar e orientar sua aplicação. Os vírus entéricos, dentre os quais se inclui o gênero Enterovirus, são potenciais contaminantes microbianos presentes no lodo de esgoto. Esses organismos são capazes de sobreviver por meses em águas e solos e sua presença no ambiente pode trazer prejuízos à saúde da população exposta aos mesmos. O objetivo do presente estudo foi de analisar a ocorrência de Enterovirus em amostras de lodo de esgoto de seis diferentes ETEs do Estado de São Paulo, avaliar o desempenho do método analítico para a detecção desses organismos e verificar se as concentrações médias obtidas atendem à legislação. Foram coletadas um total de 35 amostras no período de fevereiro de 2009 a dezembro de 2009. As análises dos Enterovirus foram realizadas pelo ensaio de plaqueamento em cultura de células RD segundo método EPA /625/R-092/013. Os resultados obtidos foram que Enterovirus estiveram presentes em 83 por cento das amostras analisadas, com concentrações que variaram de não detectado (ND) a 12,50 UFP/gST. As taxas de recuperação obtidas variaram de 0,20 por cento a 68,50 por cento . Conclui-se que das amostras analisadas 31 por cento atendem o estabelecido pela Resolução CONAMA 375/2006.

High rate of viral identification and coinfections in infants with acute bronchiolitis

OBJECTIVES: To determine the viruses and risk factors associated with hospital and intensive care unit (ICU) admissions in infants with acute bronchiolitis. INTRODUCTION: Bronchiolitis is a major cause of morbidity in infants. Widespread use of molecular-based methods has yielded new insights about its etiology, but the impact of viral etiologies on early outcomes is still unclear. METHODS: Seventy-seven infants with bronchiolitis who were under two years of age and visited an emergency unit were included. Using molecular-based methods, samples were tested for 12 different respiratory viruses. Logistic regression models were used to identify clinical and virological variables associated with the main endpoints: hospital admission and ICU admission. RESULTS: We identified at least one virus in 93.5 percent of patients, and coinfections were found in nearly 40 percent of patients. RSV was the most common pathogen (63.6 percent), followed by rhinovirus (39 percent). Identification of RSV was only associated with an increased risk of hospital admission in the univariate model. Younger age and enterovirus infection were associated with an increased risk of hospital admission, while atopy of a first-degree relative showed a protective effect. Prematurity was associated with an increased risk of admission to the ICU. Coinfections were not associated with worse outcomes. CONCLUSIONS: Molecular-based methods resulted in high rates of viral identification but did not change the significant role of RSV in acute bronchiolitis. Younger age and enterovirus infection were risk factors for hospital admission, while prematurity appeared to be a significant risk factor for admission to the ICU in acute viral bronchiolitis.

Brote de meningitis por enterovirus en el Distrito Metropolitano? / An outbreak of enterovirus meningitis in the Metropolitan District?

Los Enterovirus, agentes más frecuentes de meningitis virales, circulan durante todo el año en los países del trópico, pudiendo ocasionar un aumento en el número de casos esperados. Confirmar la existencia de un brote de meningitis viral en el Distrito Metropolitano. Descripción de las características demográficas y epidemiológicas de los niños de 1 mes a 12 años que egresaron con diagnóstico de meningitis aséptica, entre marzo y agosto de 2007, de los hospitales Universitario de Caracas y Elías Toro. Un total de 195 niños con edad promedio de 6.05 años y procedentes de 14 parroquias del Distrito Metropolitano cumplieron con los criterios (142 del HUC y 53 HET). Las características clínicas y paraclínicas de los niños coincidieron con lo reportado en la literatura. El núcleo familiar estuvo conformado en promedio por 3 adultos y 3 niños. El consumo de agua fue directamente del chorro o pipotes en 37 por ciento de las familias. El 63 por ciento restante refirieron agua hervida, filtrada o comercializada. En 2 niños se registró el inicio de los síntomas dentro de los 7 días del retorno al área Metropolitana. De 261 punciones realizadas, se estudiaron 54 en LCR para aislamiento viral: 17 con Enterovirus no polio, 4 Echovirus 30 y dos muestras con Echovirus 25. Se demostró un brote de meningitis viral en el Distrito Metropolitano, causado por Echovirus 25 y 30. Mantener informado al médico ante situaciones de brote, a fin de disminuir la transmisión del virus.

Management of enteroviral conjunctivitis outbreaks in the Singapore military in 2005.

Between 8 August and 9 October 2005, tropical Singapore experienced a national epidemic of acute viral conjunctivitis. We report the epidemiological and virological findings of outbreaks of viral conjunctivitis in military facilities during the same time period, and the outbreak control measures taken. Outbreaks of viral conjunctivitis were identified by medical officers in military medical facilities. Epidemiological and virological investigations were carried out, and a standard set of control measures was instituted for each outbreak upon detection. Eight outbreaks of viral conjunctivitis occurred in the time period, corresponding to the national outbreak. Delayed detection and institution of preventive measures during the outbreaks were associated with delayed effectiveness of the control measures. Every delay in outbreak detection by one day resulted in a delay in terminating the outbreak by 0.82 days (p = 0.01). Coxsackievirus A24 was identified from some of the conjunctival swab samples taken.

Hand, foot and mouth disease in Nagpur.

Hand, foot and mouth disease (HFMD) is a viral infection of children caused by Coxsackie virus-A16, a type of enterovirus closely related with the virus that causes herpangina. Although seen worldwide, it is not common in India. Hand, foot and mouth disease is sporadically reported from India as a mild illness. This report describes four cases of HFMD from Nagpur, Central India, seen between September 2005 and April 2006. All patients presented with a mild febrile prodrome followed by the appearance of aphthous-like oral ulcers and vesicular lesions on the hands and feet. All cases were clinically diagnosed. Coxsackie virus A16 was isolated from the serum of one of the patients. All the patients were in the age group of 3-5 years from different schools. It was a mild illness and all the four patients recovered without any complication. There were no secondary cases in the families.

Laboratory results in ocular viral diseases: implications in clinical-laboratory correlation

PURPOSE: To document etiology and predictive value of clinical diagnosis in laboratory confirmed viral diseases. METHODS: Reports of culture-positive cases of samples collected from patients presenting from January 1987 - December 2001 were evaluated. RESULTS: One thousand nine hundred and sixty-four (1964) cultures were submitted during 1987-2001. Twenty-six percent were positive (514). Human herpesvirus 1 was the most frequent agent isolated from all positive culture (56 percent). Adenovirus was the most common virus isolated from conjunctiva (66 percent), human herpesvirus 1 from lid and cornea (76 percent, 88 percent) and cytomegalovirus from vitreous (27 percent). Some unusual pathogens were recovered from conjunctiva as cytomegalovirus and from cornea as adenovirus, enterovirus and cytomegalovirus. Recognition of common viral syndromes was human herpesvirus 1 (88 percent), epidemic keratoconjunctivitis (88 percent), acute hemorrhagic conjunctivitis (70 percent) and varicella zoster virus (100 percent). However, some misdiagnosed cases were observed. Thirteen percent of conjunctivitis thought to be caused by herpes were due to adenovirus, 3.2 percent to Enterovirus, 3.2 percent to varicella zoster virus and 3.2 percent to human cytomegalovirus. Also, 5 percent of cases with a clinical diagnosis of herpes keratitis were caused by adenovirus and 2.7 percent by enterovirus. Finally, 4.8 percent of cases thought to be adenovirus conjunctivitis were herpes conjunctivitis. CONCLUSIONS: Human herpesvirus 1 remains the most frequently isolated virus from ocular sites in general (56 percent). Nonherpetic corneal isolates were in decreasing order: adenovirus, enterovirus and cytomegalovirus. Clinical and laboratory correlation was less than 90 percent. The most misdiagnosed cases were herpes conjunctivitis and keratitis, some cases of adenovirus conjunctivitis some cases of acute hemorrhagic conjunctivitis. It is essential that a rapid and...

OBJETIVOS: Documentar a etiologia e prever a importância do diagnóstico clínico em doenças virais oculares confirmadas em laboratório. MÉTODOS: Todos os relatórios de pacientes com cultura viral positiva durante o período de janeiro 1987 - dezembro 2001 foram analisados. RESULTADOS: Quinhentos e quatorze (514) casos foram encontrados. Em geral, herpesvírus 1 humano foi o agente mais freqüentemente isolado. Adenovírus foi o vírus mais comumente isolado da conjuntiva (66 por cento), herpesvírus 1 humano das pálpebras e córnea (76 por cento, 88 por cento respectivamente) e citomegalovírus do vítreo (27 por cento). Alguns agentes não usuais foram isolados da conjuntiva como citomegalovírus e da córnea como adenovírus, enterovírus e citomegalovírus. Reconhecimento das síndromes virais comuns foi: herpervírus 1 humano (88 por cento), ceratoconjuntivite epidêmica (88 por cento), conjuntivite aguda hemorrágica (70 por cento). Porém, alguns casos com diagnóstico incorreto foram observados. Treze por cento das conjuntivites com diagnóstico de herpes foram causadas por adenovírus, 3,2 por cento por enterovírus, 3,2 por cento por varicella zoster vírus e 3,2 por cento por citomegalovírus. Também, 5 por cento dos casos com diagnóstico clínico de ceratite herpética eram adenovírus e 2,7 por cento enterovírus. Além disso, 4,8 por cento dos casos em que se pensaram em conjuntivite por adenovírus, eram conjuntivite por herpes. Finalmente, 30 por cento dos casos em que se diagnosticaram conjuntive hemorrágica aguda, o agente etiológico era adenovírus. CONCLUSÃO: Em geral herpesvírus humano 1 continua a ser o vírus mais comum encontrado nas infecções oculares (56 por cento). Agentes não herpéticos isolados da córnea foram em ordem decrescente: adenovírus, enterovírus e citomegalovírus. A correlação entre o diagnóstico clínico e laboratorial foi menor do que 90 por cento. Um diagnóstico rápido e específico é essencial em casos de apresentações virais atípicas...

Epidemiología de los enterovirus asociados a enfermedades neurológicas / Epidemiology of enterovirus associated with neurologic diseases

El presente estudio describe los resultados de la investigación de los enterovirus humanos (HEV) mediante cultivo celular y reacción en cadena de la polimerasa y su tipificación molecular en 2167 casos de parálisis fláccida aguda, meningitis aséptica y encefalitis aguda, obtenidos entre 1991 y 1998 en la Argentina. La frecuencia de detección de HEV en parálisis fláccida aguda fue 19.5% (130/666) y de poliovirus Sabin 5.4% (36/666). La tasa de detección de HEV en los casos de meningitis fue 28.8% (231/801) y en encefalitis 3.0% (21/700). El grupo etario más afectado por las meningitis fue entre 1 y 9 años (75.3%) y en los casos de parálisis fláccida aguda, de 1 a 4 años (58%). En muestras de brotes de meningitis se identificó echovirus (E) 4, E9, E30 y E17, y en casos esporádicos virus coxsackie A (CAV) 2, B (CBV) 2 y CBV5, E7, E11, E19, E24 y E29, y enterovirus (EV) 71. Finalmente, en casos de encefalitis se detectó E4, E7 y E24. En casos de parálisis fláccida aguda se identificaron 28 serotipos distintos de enterovirus no polio. En la Argentina y en otros países latinoamericanos existe escasa información acerca de la circulación de los HEV y su relación con diversas enfermedades neurológicas. Este estudio proporciona información que puede servir como base para posteriores investigaciones.

This report describes the results of human enterovirus (HEV) detection and characterization using cell culture, polymerase chain reaction and molecular typing in 2167 samples obtained from acute flaccid paralysis, aseptic meningitis and acute encephalitis patients, from 1991 to 1998 in Argentina. HEV were isolated in 130 out of 666 cases (19.5%) and 36 out of 666 (5.4%). HEV RNA was detected in 28.8% (231/801) and 3.0% (21/700) of the patients with meningitis and encephalitis, respectively. Children with ages ranging from 1 to 9 years accounted for 75.3% of the meningitis cases and from 1 to 4 years for 58% of acute flaccid paralysis patients. Echovirus 4 (E4), E9, E30 and E17 were identified from meningitis outbreaks. Coxsackievirus A2 (CAV2), CBV2, CBV5, E7, E11, E19, E24, E29 and enterovirus 71 were recovered only from sporadic cases. Three different serotypes were identified in encephalitis patients: E4, E7 and E24. A total of 28 different serotypes of non-polio enteroviruses were detected from acute flaccid paralysis cases. The information here presented contributes to improving our knowledge about enteroviruses epidemiology in Argentina and their relationship with different neurological diseases. This study provides valuable data that could be useful to further research.