Effects of Tanreqing Capsule on the negative conversion time of nucleic acid in patients with COVID-19: A retrospective cohort study / 中西医结合学报

OBJECTIVE@#Traditional Chinese medicine plays a significant role in the treatment of the pandemic of coronavirus disease 2019 (COVID-19). Tanreqing Capsule (TRQC) was used in the treatment of COVID-19 patients in the Shanghai Public Health Clinical Center. This study aimed to investigate the clinical efficacy of TRQC in the treatment of COVID-19.@*METHODS@#A retrospective cohort study was conducted on 82 patients who had laboratory-confirmed mild and moderate COVID-19; patients were treated with TRQC in one designated hospital. The treatment and control groups consisted of 25 and 57 cases, respectively. The treatment group was given TRQC orally three times a day, three pills each time, in addition to conventional Western medicine treatments which were also administered to the control group. The clinical efficacy indicators, such as the negative conversion time of pharyngeal swab nucleic acid, the negative conversion time of fecal nucleic acid, the duration of negative conversion of pharyngeal-fecal nucleic acid, and the improvement in the level of immune indicators such as T-cell subsets (CD3, CD4 and CD45) were monitored.@*RESULTS@#COVID-19 patients in the treatment group, compared to the control group, had a shorter negative conversion time of fecal nucleic acid (4 vs. 9 days, P = 0.047) and a shorter interval of negative conversion of pharyngeal-fecal nucleic acid (0 vs. 2 days, P = 0.042). The level of CD3@*CONCLUSION@#Significant reductions in the negative conversion time of fecal nucleic acid and the duration of negative conversion of pharyngeal-fecal nucleic acid were identified in the treatment group as compared to the control group, illustrating the potential therapeutic benefits of using TRQC as a complement to conventional medicine in patients with mild and moderate COVID-19. The underlying mechanism may be related to the improved levels of the immune indicator CD3

Manifestaciones gastrointestinales de pacientes infectados con el nuevo Coronavirus SARS-CoV-2 / Gastrointestinal manifestations in infected patients with the novel Coronavirus SARS-CoV-2

The pandemic of the novel coronavirus SARS-CoV-2 has caused more than 2.5 millions of confirmed cases worldwide, with a mortality rate around 7%. Most cases appear to be mild with the most common symptoms being fever, dry cough, myalgia, fatigue, and dyspnea but can result in more severe disease. Also, there are described gastrointestinal manifestations: anorexia 27%, diarrhea 12%, nausea/vomiting 10%, and abdominal pain 9%. In Chile, diarrhea has been described in 7.3%, and abdominal pain 3.7%. The primary mechanism proposed is the entry of SARS-CoV-2 into the enterocytes binding the Angiotensin-converting enzyme 2 (ACE2), which is widely distributed among the gastrointestinal tract. Also, SARS-CoV-2 can cause elevated liver enzymes in up to 40% of cases. This alteration could also be caused by the binding to the ACE2 receptor in hepatocytes and cholangiocytes. Gastrointestinal symptoms and elevated liver enzymes are more frequent in severe cases. Viral RNA has been detected in stools, which may suggest a possible fecal-oral transmission. It is essential to pay attention to gastrointestinal manifestations because they are frequent and more prevalent in severe cases. The presence of viral RNA in stool forces us to take preventive measures faced with a possible mechanism of fecal-oral transmission, mainly in the gastroenterology and endoscopy services.

La pandemia por el nuevo coronavirus SARS-CoV-2 ha causado más de 2,5 millones de casos confirmados en el mundo, con una mortalidad cercana al 7%. La mayoría de los casos parecen ser leves y los síntomas más comunes son fiebre, tos seca, mialgia, fatiga y disnea, pero pueden provocar una enfermedad más grave. También se han descrito síntomas gastrointestinales: anorexia 27%, diarrea 12%, náuseas/vómitos 10%, y dolor abdominal 9%. En Chile, se ha descrito diarrea en 7,3%, y dolor abdominal 3,7%. El principal mecanismo fisiopatológico propuesto es el ingreso al enterocito utilizando la Enzima Convertidora de Angiotensina 2 (ECA2) como receptor, el cual es ampliamente distribuido a lo largo del tracto gastrointestinal. También se ha descrito alteración de pruebas hepáticas hasta en 40% de los casos. Esta alteración también podría ser causada por el ingreso a los hepatocitos y colangiocitos mediante la ECA2. Los síntomas gastrointestinales, y las alteraciones de pruebas hepáticas son más frecuentes en los pacientes graves. Se ha detectado la presencia de ARN viral en deposiciones de pacientes, lo que podría sugerir una posible vía de transmisión fecal-oral. Es importante prestar atención a las manifestaciones gastrointestinales, ya que son comunes y más frecuentes en pacientes graves. La presencia viral en las heces nos obliga a tomar medidas de precaución y prevención de un posible mecanismo de transmisión fecal-oral, especialmente en los servicios de gastroenterología y endoscopia.

COVID-19 and the gastrointestinal tract: what do we already know? / COVID-19 e o trato gastrintestinal: o que já sabemos?

ABSTRACT The new coronavirus disease pandemic is defining 2020, with almost 17.5 million infected individuals and 700 thousand deaths up to beginning of August. It is caused by SARS-CoV-2 and the transmission is through the respiratory tract. Those infected may be asymptomatic, present typical symptoms (fever, dry cough and dyspnea), gastrointestinal symptoms (diarrhea, nausea, vomiting and abdominal pain) and viral RNA in stools. The objective of this work was to review the literature related to the prevalence of gastrointestinal symptoms, and to check the possibility of fecal-oral transmission. We searched PubMed® database on COVID-19 and gastrointestinal tract and selected articles using the PRISMA method. We eliminated articles based on titles and abstracts, small number of patients and the mechanism of infection, leaving 14 studies. Comorbidities and laboratory alterations (elevation of hepatic aminotransferases and bilirubin) were related to worsening of the disease. The prevalence of gastrointestinal symptoms ranged from 6.8% to 61.3%, including diarrhea (8.14% to 33.7%), nausea/vomiting (1.53% to 26.4%), anorexia (12.1% to 40.0%) and abdominal pain (0% to 14.5%). The presence of viral RNA in stools was rarely tested, but positive in 0% to 48.1%. The gastrointestinal tract is affected by COVID-19, causing specific symptoms, laboratory alterations and viral presence in the feces. However, the results of prevalence and possibility of fecal-oral transmission were varied, requiring further studies for more assertive conclusions. It is important that healthcare professionals draw attention to this fact, since these changes can help make diagnosis and initiate early treatment.

RESUMO Com quase 17,5 milhões de infectados e 700 mil mortos até o início de agosto no mundo, a pandemia do novo coronavírus está marcando o ano de 2020. O agente causador da doença é o vírus SARS-CoV-2, e a transmissão é por via respiratória. Os infectados podem ser assintomáticos, apresentar sintomas típicos (febre, tosse seca e dispneia), sintomas gastrintestinais (diarreia, náusea, vômito e dor abdominal) e RNA viral nas fezes. O objetivo deste trabalho foi revisar a literatura relacionada com a prevalência dos sintomas gastrintestinais, e verificar se é possível a transmissão fecal-oral da doença. Fizemos uma pesquisa na base de dados PubMed® sobre a COVID-19 e o trato gastrintestinal, selecionando artigos pelo método PRISMA. Eliminamos artigos com base em títulos e resumos, quantidade pequena de pacientes e sobre mecanismo de infecção, restando 14 estudos. Comorbidades e alterações laboratoriais (elevação de aminotransferases hepáticas e bilirrubina) foram relacionadas com piora da doença. A prevalência de sintomas gastrintestinais variou entre 6,8% e 61,3%, sendo eles diarreia (8,14% a 33,7%), náusea/vômito (1,53% a 26,4%), anorexia (12,1% a 40,0%) e dor abdominal (0% a 14,5%). A presença do RNA viral foi pouco testada, mas foi positiva entre 0% a 48,1%. O trato gastrintestinal é muito acometido pela COVID-19, provocando sintomas específicos, alterações laboratoriais e presença viral nas fezes. Contudo, os resultados de prevalência e a possibilidade de transmissão fecal-oral foram variados, necessitando de estudos maiores para conclusões mais assertivas. É importante a atenção de profissionais da saúde a isso, visto que essas alterações podem ajudar no diagnóstico e a iniciar tratamento precoce.

Síntomas gastrointestinales en la enfermedad por COVID-19 y sus implicaciones en la Enfermedad Inflamatoria Intestinal / Gastrointestinal symptoms in COVID-19 and its implications in Inflammatory Bowel Disease

La pandemia por SARS-Cov-2, ha tomado gran relevancia por su impacto en los diversos sistemas de sa-lud en el mundo. Inicialmente solo se contemplaba la importancia de los síntomas respiratorios y la fiebre; sin embargo, a diario tenemos más reportes y publicaciones sobre la relevancia que está teniendo en el sistema digestivo, ya que algunos pacientes informan síntomas gastrointestinales como diarrea, vómito y dolor abdominal. Los estudios han identificado el ARN del SARS-CoV-2 en muestras de heces de pacientes infectados, en las que se encontró que su receptor que es el de la enzima convertidora de angiotensina 2 (ECA2) se expresaba altamente en las células epiteliales gastrointestinales. Esto sugiere que el SARS-CoV-2 puede infectar activamente y replicarse en el tracto gastrointestinal. Esto tiene implicaciones importantes para el tratamiento de la enfermedad, la transmisión y el control de infecciones. En pacientes con patologías crónicas como la enfermedad inflamatoria intestinal (EII), se han generado gran cantidad de preguntas e incertidumbres ante la presencia de COVID-19 junto con las implicaciones que pueden tener tanto en la severidad de los síntomas, como en la descompensación de la patología de base y en la continuidad del tratamiento inmunosupresor.(AU)

The SARS-Cov-2 pandemic has taken great relevance due to its diverse impact in the worldwide health systems. Initially, only the importance or respiratory symptoms and fever was considered, however daily we have more reports and publications about the relevance that gastrointestinal symptoms, like diarrhea, vomit, and abdominal pain, are having in COVID-19. Several studies have identified SARS-CoV-2 RNA in stool samples of infected patients, and it was also found that the viral receptor, the angiotensin-converting enzyme 2 (ACE2) receptor is highly express in the gastrointestinal cells. These findings suggest that SARS-Cov-2 can actively infect and replicate in the gastrointestinal tract. All the above have relevant implications in the disease treatment, transmission, and infection control. In patients with chronic pathologies, such as inflammatory bowel disease (IBD), many questions and uncertainties, about symptoms severity, disease decompensation and use of immunosuppressive drugs, have been generated in the presence of COVID-19.(AU)

Manifestaciones gastrointestinales de la infección por el nuevo coronavirus / Gastrointestinal features of the new coronavirus infection

El 11 de marzo del 2020 la Organización Mundial de la Salud (OMS) declaró la pandemia por el brote de la enfer-medad por coronavirus 2019 (COVID-19). Los primeros datos acerca de las manifestaciones clínicas provienen de estudios retrospectivos de Wuhan, China, epicentro de la pandemia; además del compromiso respiratorio, se ha descrito la afectación del sistema gastrointestinal. Aunque no está del todo claro el porqué del tropismo de COVID-19 por el tracto gastrointestinal, se ha demostrado que la enzima convertidora de angiotensina 2 (ECA2), la cual tiene una alta expresión en el sistema gastrointestinal, es empleada como receptor de entrada del virus. Se ha logrado documentar la posibilidad de trasmisión fecal-oral luego de demostrar la existencia del virus en las heces, incluso hasta dentro de 7-12 días después de la conversión negativa en la muestra faríngea, inde-pendientemente de la presencia o no de síntomas gastrointestinales. La afectación del sistema gastrointestinal en pacientes con infección por COVID-19 no es infrecuente; los datos reportan que los pacientes con síntomas gastrointestinales tienen un peor pronóstico. El conocimiento de estas manifestaciones nos permite aumentar la sospecha clínica y, de esta manera, establecer diagnósticos tempranos.(AU)

The World ́s Heath Organization (WHO) declared the new coronavirus infection COVID-19 a pandemic in March 11 2020. The first data about the disease clinical features were reported from Wuhan China the epi-center of the outbreak; aside from the respiratory involvement, gastrointestinal manifestations have been described. Even though the exact mechanism for the new coronavirus tropism for the gastrointestinal tract is not well understood, angiotensin converting enzyme 2 receptor seems to play an important role as the entry site for the virus, since it is widespread located throughout the enteric mucosa. Fecal oral transmission is a possible contagion route, regardless of the presence of gastrointestinal symptoms, the virus has been isolated from feces even 7-12 days after a negative pharyngeal swab. COVID's-19 gastrointestinal involvement is not an infrequent event, data reports patients who present with these symptoms have worst prognosis. Knowing the possible gastrointestinal manifestations might rise our clinical suspicion in order to establish earlier and timely diagnosis.(AU)

Expressão e atividade de sistemas de dois componentes (TCS) de Salmonella enterica sorovar Typhimurium no contexto do metaboloma intestinal humano / Expression and activity of two-component systems of Salmonella enterica sorovar Typhimurium in the context of the human intestinal metabolome

A interação entre membros do microbioma intestinal humano, células hospedeiras e patógenos invasores pode ocorrer de diversas formas, sendo uma delas através de pequenas moléculas chamadas metabólitos. A percepção e resposta efetiva de um microrganismo às diferentes condições encontradas em seu ambiente, incluindo metabólitos produzidos por outros microrganismos, são fatores importantes para sua adaptação, sobrevivência e disseminação. Os sistemas de dois componentes (TCS) permitem a percepção e resposta a mudanças ambientais, regulando a expressão de genes específicos. Nosso grupo mostrou anteriormente que um extrato orgânico de fezes humanas (EF), bem como o ácido 3,4-dimetilbenzoico (3,4-DMB), encontrado no EF, inibe a capacidade de Salmonella enterica sorovar Typhimurium de invadir células hospedeiras. O presente trabalho propôs investigar o impacto do microbioma intestinal humano, bem como de pequenas moléculas produzidas por Clostridium citroniae (membro deste microbioma) na expressão e atividade dos genes de TCS de Salmonella. Os metabólitos de EF e de culturas puras de C. citroniae foram extraídos com acetato de etila e adicionados a meio de cultura. O pH do meio foi ajustado (~ 7,4) e a solução foi esterilizada por filtragem. Salmonella foi cultivada na presença ou ausência do EF e do extrato de C. citroniae, bem como do ácido 3,4-DMB, em condições aeróbias e anaeróbias, até alcançar o meio da fase logarítmica de crescimento. O RNA foi extraído para a realização de PCR em Tempo Real utilizando iniciadores direcionados a quase todos os TCS de Salmonella. Nossos resultados mostraram que vários genes de TCS envolvidos na virulência de Salmonella (SsrAB, EnvZ-OmpR, QseCB, PhoQP, TorSR, TtrRS) foram regulados diferencialmente por esses metabólitos, tanto em condições aeróbias quanto anaeróbias. EnvZ-OmpR, PhoPQ e SsrAB estão diretamente envolvidos na regulação das Ilhas de Patogenicidade 1 e 2 de Salmonella. QseCB é crucial para a detecção de quorum em Salmonella, de hormônios hospedeiros e para a regulação da motilidade (swimming). Vários outros TCS também foram regulados, incluindo TorSR e TtrRS, envolvidos na regulação da respiração anaeróbica de N-óxido de trimetilamina (TMAO) e tetrationato, respectivamente. Esses compostos são importantes para a sobrevivência de Salmonella no ambiente anaeróbico do intestino humano. Nossos resultados de avaliação de expressão gênica global de Salmonella cultivada na presença de ácido 3,4-DMB (aerobiose e anaerobiose) bem como na presença do EF em anaerobiose, mostraram que genes condificados em SPI-1 e SPI-2, SPI-4 e alguns genes do TCS foram reprimidos, enquanto genes marR, marB e marA foram ativadas nessas condições. Adicionalmente, comparamos nossos resultados de RNAseq, de Salmonella cultivada na presença do ácido 3,4-DMB em aerobiose, com resultados disponíveis da base de dados Salmonella Compendium. Ainda, a capacidade de Salmonella de adentrar e sobreviver dentro de células fagocíticas (macrófagos RAW 264.7) parece ser afetada pelas três condições testadas neste trabalho. Nossos resultados mostram que importantes vias de sinalização da virulência de Salmonella podem ser moduladas pelos metabólitos presentes no microbioma intestinal humano e abrem caminhos para novas pesquisas sobre a sinalização intercelular microbioma-patógeno no ambiente intestinal.

The interaction between members of the human gut microbiome, host cells and invading pathogens often occurs through small molecules, also called metabolites. The perception and effective response of a microorganism to the different conditions found in its environment, including metabolites produced by other microbes, is important for its adaptation, survival and dissemination. Two-component systems (TCS) allow the perception and response to environmental changes by regulating the expression of specific genes. Our group previously showed that organic extracts of human feces (EF) as well as the specific metabolite 3,4-dimethylbenzoic acid (3,4-DMB) found within the EF, inhibit the ability of Salmonella enterica sorovar Typhimurium to invade host cells. In the present work, we investigated the impact of the human gut microbiome as well as small molecules produced by Clostridium citroniae (a member of this microbiome) on the expression and activity of Salmonella TCS genes. Metabolites (from feces or C. citroniae cultures) were extracted using ethyl acetate and added to culture medium. The pH of the medium was adjusted (~7.4), and the solution was filter sterilized. Salmonella was grown in the presence or absence of the organic extracts as well as 3,4-DMB acid under aerobic and anaerobic conditions until it reached mid-log growth. RNA was then extracted for Real-time PCR using primers targeting almost all Salmonella TCS. Our results showed that several TCS involved in Salmonella virulence (SsrAB, EnvZ-OmpR, QseCB, PhoQP, TorSR, TtrRS) were differentially regulated by these metabolites both in aerobic and anaerobic conditions. EnvZ-OmpR, PhoPQ, and SsrAB are directly involved in the regulation of Salmonella Pathogenicity Islands 1 and 2. QseCB is crucial for Salmonella =quorum sensing, sensing of host hormones and regulation of swimming motility. Several other TCS were also regulated, including TorSR and TtrRS, which are involved in the anaerobic respiration of trimethylamine N-oxide (TMAO) and tetrathionate, respectively. These compounds are important for Salmonella survival in the anaerobic environment of the human gut. Our results of the evaluation of global Salmonella gene expression grown in the presence of 3,4-DMB acid (aerobiosis and anaerobiosis) as well as in the presence of EF in anaerobiosis, showed that genes encoded in SPI-1 and SPI-2, SPI-4 and some TCS genes have been repressed, while multiple drug resistance genes, as well marR, marB and marA genes have been activated under these conditions. Besides, we compared our results of RNAseq, Salmonella was grown in the presence of 3,4-DMB acid in aerobiosis, with results available from the Salmonella Compendium database. Also, Salmonella's ability to enter and survive within phagocytic cells (macrophages RAW 264.7) appears to be affected by the three conditions tested in this work. Our results show that important Salmonella virulence signalling pathways can be modulated by the metabolites present in the human intestinal microbiome and open the way for further research on the microbiome-pathogen intercellular signalling in the intestinal environment.

The rare enterovirus c99 and echovirus 29 strains in Brazil: potential risks associated to silent circulation

Human enteroviruses (EVs) are associated with a wide spectrum of human diseases. Here we report the complete genome sequences of one EV-C99 strain and one E29 strain obtained from children suffering from acute gastroenteritis, without symptoms of enteroviral syndromes. This is the first report of EV-C99 in South America, and the second E29 genome described worldwide. Continuous surveillance on EVs is vital to provide further understanding of the circulation of new or rare EV serotypes in the country. The present study also highlights the capacity of EVs to remain in silent circulation in populations.

Evolutionary study of potentially zoonotic hepatitis E virus genotype 3 from swine in Northeast Brazil

Hepatitis E virus (HEV), an emerging virus associated with acute hepatic disease, leads to thousands of deaths worldwide. HEV has already been reported in Brazil; however, there is a lack of epidemiological and molecular information on the genetic variability, taxonomy, and evolution of HEV. It is thus unclear whether hepatitis E is a neglected disease in Brazil or it has low relevance for public health in this country. Here, for the first time, we report the presence of HEV in Northeast Brazil. A total of 119 swine faecal samples were screened for the presence of HEV RNA using real-time polymerase chain reaction (RT-PCR) and further confirmed by conventional RT-PCR; among these, two samples were identified as positive. Molecular evolution analyses based on capsid sequences revealed that the samples had close proximities to HEV sequences belonging to genotype 3 and were genetically related to subtype 3f isolated in humans. Parsimony ancestral states analysis indicated gene flow events from HEV cross-species infection, suggesting an important role of pig hosts in viral spillover. HEV's ability for zoonotic transmission by inter-species host switching as well as its possible adaptation to new animal species remain important issues for human health.

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.

Detection and genetic characterization of Mamastrovirus 5 from Brazilian dogs

Abstract Mamastrovirus 5 (MAstV5), belonging to the Astroviridae (AstV) family, previously known as canine astrovirus or astrovirus-like particles, has been reported in several countries to be associated with viral enteric disease in dogs since the 1980s. Astroviruses have been detected in fecal samples from a wide variety of mammals and birds that are associated with gastroenteritis and extra enteric manifestations. In the present study, RT-PCR was used to investigate the presence of MAstV5 in 269 dog fecal samples. MAstV5 was detected in 26% (71/269) of the samples. Interestingly, all MAstV5-positive samples derived from dogs displaying clinical signs suggestive of gastroenteritis, other enteric viruses were simultaneously detected (canine parvovirus, canine distemper virus, canine coronavirus, canine adenovirus and canine rotavirus). Based on genomic sequence analysis of MAstV5 a novel classification of the species into four genotypes, MAstV5a-MAstV5d, is proposed. Phylogenetic analyses based on the ORF2 amino acid sequences, samples described herein grouped into the putative genotype 'a' closed related with Chinese samples. Other studies are required to attempt the clinical and antigenic implications of these astrovirus genotypes in dogs.

Identification of co-infection by rotavirus and parvovirus in dogs with gastroenteritis in Mexico

ABSTRACT This is the first report on circulating canine rotavirus in Mexico. Fifty samples from dogs with gastroenteritis were analyzed used polymerase chain reaction and reverse transcription polymerase chain reaction in order to identify parvovirus and rotavirus, respectively; 7% of dogs were infected with rotavirus exclusively, while 14% were co-infected with both rotavirus and parvovirus; clinical signs in co-infected dogs were more severe.

High prevalence of norovirus in children with sporadic acute gastroenteritis in Manaus, Amazon Region, northern Brazil

BACKGROUND Norovirus (NoV) is a major cause of acute gastroenteritis (AGE) worldwide, especially in children under five years. Studies involving the detection and molecular characterisation of NoV have been performed in Brazil, demonstrating its importance as an etiological agent of AGE. OBJECTIVES The objectives of this study were to investigate the frequency of human NoV and to genotype the strains isolated from 0-14-year-old patients of AGE in Manaus, Brazil, over a period of two years. METHODS A total of 426 faecal samples were collected between January 2010 and December 2011. All samples were tested for the presence of NoV antigens using a commercial enzyme immunoassay kit. RNA was extracted from all faecal suspensions and reverse transcription-polymerase chain reaction (RT-PCR) for the NoV-polymerase partial region was performed as a trial test. Positive samples were then subjected to PCR with specific primers for partial capsid genes, which were then sequenced. FINDINGS NoV was detected in 150 (35.2%) faecal samples, for at least one of the two techniques used. NoV was detected in children from all age groups, with the highest positivity observed among the group of 1-2 years old. Clinically, fever was verified in 43% of the positive cases and 46.3% of the negative cases, and vomiting was observed in 75.8% and 70.8% cases in these groups, respectively. Monthly distribution showed that the highest positivity was observed in January 2010 (81.2%), followed by February and April 2010 and March 2011, when the positivity rate reached almost 50%. Phylogenetic analyses performed with 65 positive strains demonstrated that 58 (89.2%) cases of NoV belonged to genotype GII.4, five (7.7%) to GII.6, and one (1.5%) each to GII.7 and GII.3. MAIN CONCLUSIONS This research revealed a high circulation of NoV GII.4 in Manaus and contributed to the understanding of the importance of this virus in the aetiology of AGE cases, especially in a region with such few studies available.

High frequency of hepatitis E virus infection in swine from South Brazil and close similarity to human HEV isolates

Abstract Hepatitis E virus is responsible for acute and chronic liver infections worldwide. Swine hepatitis E virus has been isolated in Brazil, and a probable zoonotic transmission has been described, although data are still scarce. The aim of this study was to investigate the frequency of hepatitis E virus infection in pigs from a small-scale farm in the rural area of Paraná State, South Brazil. Fecal samples were collected from 170 pigs and screened for hepatitis E virus RNA using a duplex real-time RT-PCR targeting a highly conserved 70 nt long sequence within overlapping parts of ORF2 and ORF3 as well as a 113 nt sequence of ORF2. Positive samples with high viral loads were subjected to direct sequencing and phylogenetic analysis. hepatitis E virus RNA was detected in 34 (20.0%) of the 170 pigs following positive results in at least one set of screening real-time RT-PCR primers and probes. The swine hepatitis E virus strains clustered with the genotype hepatitis E virus-3b reference sequences in the phylogenetic analysis and showed close similarity to human hepatitis E virus isolates previously reported in Brazil.

Diversity of group A rotavirus genes detected in the Triângulo Mineiro region, Minas Gerais, Brazil

ABSTRACT Group A rotaviruses are the main causative agent of infantile gastroenteritis. The segmented nature of the viral genome allows reassortment of genome segments, which can generate genetic variants. In this study, we characterized the diversity of the VP7, VP4 (VP8*), VP6, NSP4, and NSP5 genes of the rotaviruses that circulated from 2005 to 2011 in the Triângulo Mineiro (TM) region of Brazil. Samples with genotypes G2 (sublineages IVa-1 and IVa-3), G1 (sublineage I-A), G9 (lineage III), G12 (lineages II and III), G8 (lineage II), G3 (lineage III), P[4] (sublineages IVa and IVb), P[8] (sublineages P[8]-3.6, P[8]-3.3, and P[8]-3.1), I2 (lineage VII), E2 (lineages VI, XII, and X), and H2 (lineage III) were identified. The associations found in the samples were G1, G9, or G12 with P[8]-I1-E1-H1; G2 or G8 with P[4]-I2-E2-H2; G12 with I3-E3-H6; and G3 with P[4]-I2-E3-H3 (previously unreported combination). Reassortment events in G2P[4] strains and an apparent pattern of temporal segregation within the lineages were observed. Five TM samples contained genes that exhibited high nucleotide and amino acid identities with strains of animal origin. The present study includes a period of pre- and post-introduction of rotavirus vaccination in all Brazilian territories, thereby serving as a basis for monitoring changes in the genetic constitution of rotaviruses. The results also contribute to the understanding of the diversity and evolution of rotaviruses in a global context.

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.

The role of human adenoviruses type 41 in acute diarrheal disease in Minas Gerais after rotavirus vaccination

Abstract Human adenovirus species F (HAdV-F) type 40 and 41 are commonly associated with acute diarrheal disease (ADD) across the world. Despite being the largest state in southeastern Brazil and having the second largest number of inhabitants, there is no information in the State of Minas Gerais regarding the role of HAdV-F in the etiology of ADD. This study was performed to determine the prevalence, to verify the epidemiological aspects of infection, and to characterize the strains of human adenoviruses (HAdV) detected. A total of 377 diarrheal fecal samples were obtained between January 2007 and August 2011 from inpatient and outpatient children of age ranging from 0 to 12 years. All samples were previously tested for rotavirus, norovirus, and astrovirus, and 314 of 377 were negative. The viral DNA was extracted, amplified using the polymerase chain reaction and the HAdV-positive samples were sequenced and phylogenetically analyzed. Statistical analyses were performed using the Chi-square test (p < 0.05), considering two conditions: the total of samples tested (377) and the total of negative samples for the remaining viruses tested (314). The overall prevalence of HAdV was 12.47% (47/377); and in 76.60% (36/47) of the positive samples, this virus was the only infectious agent detected. The phylogenetic analysis of partial sequences of 32 positive samples revealed that they all clustered with the HAdV-F type 41. The statistical analysis showed that there was no correlation between the onset of the HAdV infection and the origin of the samples (inpatients or outpatients) in the two conditions tested: the total of samples tested (p = 0.598) and the total of negative samples for the remaining viruses tested (p = 0.614). There was a significant association in the occurrence of infection in children aged 0–12 months for the condition 1 (p = 0.030) as well as condition 2 (p = 0.019). The occurrence of infections due to HAdV did not coincide with a pattern of seasonal distribution. These data indicate the significant involvement of HAdV-F type 41 in the etiology of ADD in Minas Gerais, which demonstrates the importance of other viral agents in the development of the disease after the introduction of rotavirus vaccine immunization.

Selection of superior bifidobacteria in the presence of rotavirus

The main purpose of this study was to investigate bifidobacteria flora in fecal samples from children with rotavirus infection and determine the significance of their selected probiotic properties for improvement of health status. Enzyme-linked immunosorbent assay was used to identify rotavirus antigen in fecal samples from 94 patients with gastroenteritis and from 30 without gastroenteritis. Bifidobacteria were identified by selective media, gram reaction, colony morphology, fructose-6-phosphate phosphoketolase enzyme activity and classical identification tests. Exopolysaccharide (EPS) production was identified by phenol-sulphuric acid method. The modified method was then used to identify the quantity of taurocholic and glycocholic acid deconjugation and cholesterol elimination of the strains. Thirty-five of the 94 fecal samples were found positive for rotavirus antigen (37.23%). Bifidobacteria were identified in 59 of the samples. The EPS production ranges were 29.56-102.21 mg/L. The cholesterol elimination rates ranged between 8.36-39.22%. Furthermore, a positive and strong correlation was determined between EPS production and the presence of cholesterol (r=0.984, P<0.001). The deconjugation rates for the sodium glycocholate group was higher than the sodium taurocholate group. Rotavirus (+) bifidobacteria strains had higher EPS production, deconjugation rate and cholesterol elimination compared to bifidobacteria strains isolated from children in the rotavirus (-) sample and without gastroenteritis. Significant differences were observed among groups in all parameters (P<0.05). Given the increased number of rotavirus cases in Turkey and worldwide, it is very important to add superior bifidobacteria in the diets of infected children to improve the intestinal and vital functions.

Neurologic Manifestations of Enterovirus 71 Infection in Korea

Enterovirus 71 frequently involves the central nervous system and may present with a variety of neurologic manifestations. Here, we aimed to describe the clinical features, magnetic resonance imaging (MRI) findings, and cerebrospinal fluid (CSF) profiles of patients presenting with neurologic complications of enterovirus 71 infection. We retrospectively reviewed the records of 31 pediatric patients hospitalized with acute neurologic manifestations accompanied by confirmed enterovirus 71 infection at Ulsan University Hospital between 2010 and 2014. The patients' mean age was 2.9 ± 5.5 years (range, 18 days to 12 years), and 80.6% of patients were less than 4 years old. Based on their clinical features, the patients were classified into 4 clinical groups: brainstem encephalitis (n = 21), meningitis (n = 7), encephalitis (n = 2), and acute flaccid paralysis (n = 1). The common neurologic symptoms included myoclonus (58.1%), lethargy (54.8%), irritability (54.8%), vomiting (48.4%), ataxia (38.7%), and tremor (35.5%). Twenty-five patients underwent an MRI scan; of these, 14 (56.0%) revealed the characteristic increased T2 signal intensity in the posterior region of the brainstem and bilateral cerebellar dentate nuclei. Twenty-six of 30 patients (86.7%) showed CSF pleocytosis. Thirty patients (96.8%) recovered completely without any neurologic deficits; one patient (3.2%) died due to pulmonary hemorrhage and shock. In the present study, brainstem encephalitis was the most common neurologic manifestation of enterovirus 71 infection. The characteristic clinical symptoms such as myoclonus, ataxia, and tremor in conjunction with CSF pleocytosis and brainstem lesions on MR images are pathognomonic for diagnosis of neurologic involvement by enterovirus 71 infection.

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..