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Sample records for astroviruses seadornaviruses cytorhabdoviruses

  1. Candidates in Astroviruses, Seadornaviruses, Cytorhabdoviruses and Coronaviruses for +1 frame overlapping genes accessed by leaky scanning

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    Atkins John F

    2010-01-01

    Full Text Available Abstract Background Overlapping genes are common in RNA viruses where they serve as a mechanism to optimize the coding potential of compact genomes. However, annotation of overlapping genes can be difficult using conventional gene-finding software. Recently we have been using a number of complementary approaches to systematically identify previously undetected overlapping genes in RNA virus genomes. In this article we gather together a number of promising candidate new overlapping genes that may be of interest to the community. Results Overlapping gene predictions are presented for the astroviruses, seadornaviruses, cytorhabdoviruses and coronaviruses (families Astroviridae, Reoviridae, Rhabdoviridae and Coronaviridae, respectively.

  2. Astrovirus Diagnostics

    Science.gov (United States)

    Pérot, Philippe; Lecuit, Marc; Eloit, Marc

    2017-01-01

    Various methods exist to detect an astrovirus infection. Current methods include electron microscopy (EM), cell culture, immunoassays, polymerase chain reaction (PCR) and various other molecular approaches that can be applied in the context of diagnostic or in surveillance studies. With the advent of metagenomics, novel human astrovirus (HAstV) strains have been found in immunocompromised individuals in association with central nervous system (CNS) infections. This work reviews the past and current methods for astrovirus detection and their uses in both research laboratories and for medical diagnostic purposes. PMID:28085120

  3. The Astrovirus Capsid: A Review

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    Arias, Carlos F.; DuBois, Rebecca M.

    2017-01-01

    Astroviruses are enterically transmitted viruses that cause infections in mammalian and avian species. Astroviruses are nonenveloped, icosahedral viruses comprised of a capsid protein shell and a positive-sense, single-stranded RNA genome. The capsid protein undergoes dramatic proteolytic processing both inside and outside of the host cell, resulting in a coordinated maturation process that affects cellular localization, virus structure, and infectivity. After maturation, the capsid protein controls the initial phases of virus infection, including virus attachment, endocytosis, and genome release into the host cell. The astrovirus capsid is the target of host antibodies including virus-neutralizing antibodies. The capsid protein also mediates the binding of host complement proteins and inhibits complement activation. Here, we will review our knowledge on the astrovirus capsid protein (CP), with particular attention to the recent structural, biochemical, and virological studies that have advanced our understanding of the astrovirus life cycle. PMID:28106836

  4. Complete genome sequence of Colocasia bobone disease-associated virus, a putative cytorhabdovirus infecting taro.

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    Higgins, Colleen M; Bejerman, Nicolas; Li, Ming; James, Anthony P; Dietzgen, Ralf G; Pearson, Michael N; Revill, Peter A; Harding, Robert M

    2016-03-01

    We report the first genome sequence of a Colocasia bobone disease-associated virus (CBDaV) derived from bobone-affected taro [Colocasia esculenta L. Schott] from Solomon Islands. The negative-strand RNA genome is 12,193 nt long, with six major open reading frames (ORFs) with the arrangement 3'-N-P-P3-M-G-L-5'. Typical of all rhabdoviruses, the 3' leader and 5' trailer sequences show complementarity to each other. Phylogenetic analysis indicated that CBDaV is a member of the genus Cytorhabdovirus, supporting previous reports of virus particles within the cytoplasm of bobone-infected taro cells. The availability of the CBDaV genome sequence now makes it possible to assess the role of this virus in bobone, and possibly alomae disease of taro and confirm that this sequence is that of Colocasia bobone disease virus (CBDV).

  5. Nosocomial diarrhea in children:is astrovirus the leading pathogen?

    Institute of Scientific and Technical Information of China (English)

    Anita Chakravarti; Manisha Jain; Mayank Singh Chauhan; Anju Sharma; Sayani Tewari

    2010-01-01

    Objective:To screen for the presence of mixed infection with rotavirus. Methods:The present study included 140 children aged less than 2 years with acute diarrhea. Fecal samples of all these patients were analyzed for the presence of astroviral antigen by enzyme immunoassay. Also 40 rotavirus positive fecal samples were screened for the presence of astrovirus. Results:In case of acute diarrhea in children the prevalence of astrovirus was around 34%(48/140). It was seen that even in rotavirus positive cases astrovirus co infection was 25%. Conclusions:Astrovirus is a growing problem which is often underrecognised. With the rotavirus vaccine licensure being imminent astrovirus will emerge out as the single most important cause of viral diarrhea.

  6. Novel Coronaviruses and Astroviruses in Bats

    Institute of Scientific and Technical Information of China (English)

    Daniel K. W. Chu; J. S. Malik Peiris; Leo L. M. Poon

    2009-01-01

    Zoonotic transmissions of emerging pathogens from wildlife to human have shaped the history of mankind. These events have also highlighted our poor understanding of microorganisms circulated in wild animals. Coronaviruses and astroviruses, which can be found from a wide range of mammals, were recently detected in bats. Strikingly, these bat viruses are genetically highly diverse and these interesting findings might help to better understand the evolution and ecology of these viruses. The discoveries of these novel bats viruses not only suggested that bats are important hosts for these virus families, but also reiterated the role of bats as a reservoir of viruses that might pose a zoonotic threat to human health.

  7. Cytorhabdovirus P3 genes encode 30K-like cell-to-cell movement proteins.

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    Mann, Krin S; Bejerman, Nicolas; Johnson, Karyn N; Dietzgen, Ralf G

    2016-02-01

    Plant viruses encode movement proteins (MP) to facilitate cell-to-cell transport through plasmodesmata. In this study, using trans-complementation of a movement-defective turnip vein-clearing tobamovirus (TVCV) replicon, we show for the first time for cytorhabdoviruses (lettuce necrotic yellows virus (LNYV) and alfalfa dwarf virus (ADV)) that their P3 proteins function as MP similar to the TVCV P30 protein. All three MP localized to plasmodesmata when ectopically expressed. In addition, we show that these MP belong to the 30K superfamily since movement was inhibited by mutation of an aspartic acid residue in the critical 30K-specific LxD/N50-70G motif. We also report that Nicotiana benthamiana microtubule-associated VOZ1-like transcriptional activator interacts with LNYV P3 and TVCV P30 but not with ADV P3 or any of the MP point mutants. This host protein, which is known to interact with P3 of sonchus yellow net nucleorhabdovirus, may be involved in aiding the cell-to-cell movement of LNYV and TVCV.

  8. Astrovirus infection in young children with diarrhea hospitalized at Beijing Children's Hospital

    Institute of Scientific and Technical Information of China (English)

    刘春艳; 申昆玲; 王树欣; 刘亚谊; 照日格图

    2004-01-01

    Background This study was to assess certain epidemiological and clinical characteristics of astrovirus infection in children with diarrhea below 5 years old hospitalized at the Beijing Children's Hospital. Methods From September 1999 to August 2001, a total of 288 children with diarrhea below 5 years old hospitalized at Beijing Children's Hospital were studied. Fecal specimens collected from each patient were tested for astrovirus by using enzyme immunoassay (EIA) and further serotyped by reverse-transcription polymerase chain reaction (RT-PCR) using serotype-specific primers. Results Astrovirus antigen was positive in 9.0% of the patients, and the rate of coinfection with rotavirus was 19.2% among the astrovirus positive patients. The patients with astrovirus infection were at the age of 7 days to 17 months, most (92.3%) of them were less than 12 months. Astrovirus infection occurred mainly between October and March of the following year. Main symptoms included watery stool, vomiting, fever, and dehydration. Increased activity of myocardial enzyme was seen in 4 patients. Of the 22 astrovirus positive specimens, 14 were successfully serotyped by RT-PCR and they were all serotype 1. Conclusions The overall incidence of astrovirus infection in young children with diarrhea hospitalized at the Beijing Children's Hospital was 9.0%. The predominant serotype was type 1. Astrovirus may play an important role in viral diarrhea in young children in Beijing, China.

  9. Crystal Structure of the Human Astrovirus Capsid Protein

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    Toh, Yukimatsu; Harper, Justin; Dryden, Kelly A.; Yeager, Mark; Méndez, Ernesto

    2016-01-01

    ABSTRACT Human astrovirus (HAstV) is a leading cause of viral diarrhea in infants and young children worldwide. HAstV is a nonenveloped virus with a T=3 capsid and a positive-sense RNA genome. The capsid protein (CP) of HAstV is synthesized as a 90-kDa precursor (VP90) that can be divided into three linear domains: a conserved N-terminal domain, a hypervariable domain, and an acidic C-terminal domain. Maturation of HAstV requires proteolytic processing of the astrovirus CP both inside and outside the host cell, resulting in the removal of the C-terminal domain and the breakdown of the rest of the CP into three predominant protein species with molecular masses of ∼34, 27/29, and 25/26 kDa, respectively. We have now solved the crystal structure of VP9071–415 (amino acids [aa] 71 to 415 of VP90) of human astrovirus serotype 8 at a 2.15-Å resolution. VP9071–415 encompasses the conserved N-terminal domain of VP90 but lacks the hypervariable domain, which forms the capsid surface spikes. The structure of VP9071–415 is comprised of two domains: an S domain, which adopts the typical jelly-roll β-barrel fold, and a P1 domain, which forms a squashed β-barrel consisting of six antiparallel β-strands similar to what was observed in the hepatitis E virus (HEV) capsid structure. Fitting of the VP9071–415 structure into the cryo-electron microscopy (EM) maps of HAstV produced an atomic model for a continuous, T=3 icosahedral capsid shell. Our pseudoatomic model of the human HAstV capsid shell provides valuable insights into intermolecular interactions required for capsid assembly and trypsin-mediated proteolytic maturation needed for virus infectivity. Such information has potential applications in the development of a virus-like particle (VLP) vaccine as well as small-molecule drugs targeting astrovirus assembly/maturation. IMPORTANCE Human astrovirus (HAstV) is a leading cause of viral diarrhea in infants and young children worldwide. As a nonenveloped virus

  10. Turkey Astrovirus Type 1 (TAstV-1) and Chicken Astrovirus (CAstV) Detection in Brazilian Chicken Flocks.

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    Espinoza, Luis Luna; Beserra, Laila A R; Soares, Rodrigo M; Gregori, Fabio

    2016-09-01

    Astrovirus is a common cause of enteritis in humans and domestic animals. Here we report the detection of turkey astrovirus type 1 (TAstV-1) and chicken astrovirus (CAstV) in avian farms. Sixty fecal sample pools (five or six birds of the same flock), from chickens without apparent clinical symptoms of enteric disease from farms located in six Brazilian states, were screened by an ORF1b PCR, followed by nucleotide sequencing of amplified products and phylogenetic analysis. Six samples tested positive for TAstV-1 and two for CAstV. One positive sample of each detected virus (TAstV-1 and CAstV) had the complete ORF2 sequenced. Data for the ORF2 sequence indicate that Brazilian TAstV-1 was divergent from TAstV-1 (United States), previously described infecting turkeys, and Brazilian CAstV clustered together with the U.K. group, subgroup B-II, associated with enteritis and growth retardation in chicks. This study provides updated information about CAstV and the first report of detection of TAstV-1 in Brazilian chickens, supporting the diagnostic of enteritis and epidemiologic surveillance in poultry health.

  11. Recombinant proteins as vaccines for protection against disease induced by infection with mink astrovirus

    DEFF Research Database (Denmark)

    2012-01-01

    and polypeptides of the capsid protein of a novel mink astrovirus strain denoted DK7627. Such polynucleotides and polypeptides may be used for the production of vaccines against mink astrovirus which may induce pre-weaning diarrhoea in minks. The invention furthermore relates to vectors, host cells, compositions...

  12. Childhood astrovirus-associated diarrhea in the ambulatory setting in a Public Hospital in Cordoba city, Argentina

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    Giordano Miguel O.

    2004-01-01

    Full Text Available Human astroviruses have been increasingly identified as important agents of diarrheal disease in children. However, the disease burden of astrovirus infection is still incompletely assessed. This paper reports results on the epidemiological and clinical characteristics of astrovirus-associated diarrhea, as well as the impact of astrovirus infection on the ambulatory setting at a Public Hospital in Córdoba city, Argentina. From February 2001 through January 2002, 97 randomly selected outpatient visits for diarrhea among children 0.05. According to our estimation about one out of seventy-four children in this cohort would be assisted annually for an astroviral-diarrheal episode in the Public Hospital and one out of eight diarrheal cases could be attributed to astrovirus infection. Astrovirus is a common symptomatic infection in pediatric outpatient visits in the public hospital in the study area, contributing 12.37% of the overall morbidity from diarrhea.

  13. Oral Administration of Astrovirus Capsid Protein Is Sufficient To Induce Acute Diarrhea In Vivo

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    Victoria A. Meliopoulos

    2016-11-01

    Full Text Available The disease mechanisms associated with the onset of astrovirus diarrhea are unknown. Unlike other enteric virus infections, astrovirus infection is not associated with an inflammatory response or cellular damage. In vitro studies in differentiated Caco-2 cells demonstrated that human astrovirus serotype 1 (HAstV-1 capsid protein alone disrupts the actin cytoskeleton and tight junction complex, leading to increased epithelial barrier permeability. In this study, we show that oral administration of purified recombinant turkey astrovirus 2 (TAstV-2 capsid protein results in acute diarrhea in a dose- and time-dependent manner in turkey poults. Similarly to that induced by infectious virus, TAstV-2 capsid-induced diarrhea was independent of inflammation or histological changes but was associated with increased intestinal barrier permeability, as well as redistribution of sodium hydrogen exchanger 3 (NHE3 from the membrane to the cytoplasm of the intestinal epithelium. Unlike other viral enterotoxins that have been identified, astrovirus capsid induces diarrhea after oral administration, reproducing the natural route of infection and demonstrating that ingestion of intact noninfectious capsid protein may be sufficient to provoke acute diarrhea. Based on these data, we hypothesize that the astrovirus capsid acts like an enterotoxin and induces intestinal epithelial barrier dysfunction.

  14. Full genome analysis of bovine astrovirus from fecal samples of cattle in Japan: identification of possible interspecies transmission of bovine astrovirus.

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    Nagai, Makoto; Omatsu, Tsutomu; Aoki, Hiroshi; Otomaru, Konosuke; Uto, Takehiko; Koizumi, Motoya; Minami-Fukuda, Fujiko; Takai, Hikaru; Murakami, Toshiaki; Masuda, Tsuneyuki; Yamasato, Hiroshi; Shiokawa, Mai; Tsuchiaka, Shinobu; Naoi, Yuki; Sano, Kaori; Okazaki, Sachiko; Katayama, Yukie; Oba, Mami; Furuya, Tetsuya; Shirai, Junsuke; Mizutani, Tetsuya

    2015-10-01

    A viral metagenomics approach was used to investigate fecal samples of Japanese calves with and without diarrhea. Of the different viral pathogens detected, read counts gave nearly complete astrovirus-related RNA sequences in 15 of the 146 fecal samples collected in three distinct areas (Hokkaido, Ishikawa, and Kagoshima Prefectures) between 2009 and 2015. Due to the lack of genetic information about bovine astroviruses (BoAstVs) in Japan, these sequences were analyzed in this study. Nine of the 15 Japanese BoAstVs were closely related to Chinese BoAstVs and clustered into a lineage (tentatively named lineage 1) in all phylogenetic trees. Three of 15 strains were phylogenetically separate from lineage 1, showing low sequence identities, and clustered instead with an American strain isolated from cattle with respiratory disease (tentatively named lineage 2). Interestingly, two of 15 strains clustered with lineage 1 in the open reading frame (ORF)1a and ORF1b regions, while they clustered with lineage 2 in the ORF2 region. Remarkably, one of 15 strains exhibited low amino acid sequence similarity to other BoAstVs and was clustered separately with porcine astrovirus type 5 in all trees, and ovine astrovirus in the ORF2 region, suggesting past interspecies transmission.

  15. Molecular characterization of a novel astrovirus associated with disease in mink

    DEFF Research Database (Denmark)

    Mittelholzer, C.; Hedlund, K.O.; Englund, L.;

    2003-01-01

    , containing sequence motifs for a serine protease (ORF1a) and an RNA-dependent RNA polymerase (ORF1b). The structural proteins are encoded by ORF2 and, presumably, are expressed as a polyprotein precursor to be cleaved into the mature capsid proteins. These results indicate that mink astrovirus (MiAstV) has...

  16. Astrovirus infection in children living in the Central West region of Brazil

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    Rodrigo Alessandro Tôgo Santos

    2007-03-01

    Full Text Available This study presents data regarding the circulation of astrovirus in Goiânia-GO and Brasília-DF. These viruses were detected in fecal samples from hospitalized children up to five years old with and without acute gastroenteritis. A total of 1244 fecal samples were collected in two periods, 1994 to 1996 (Brasília and 1998 to 2002 (Goiânia and Brasília, and were analyzed for viral RNA using the reverse transcription-polymerase chain reaction (RT-PCR. Positivity rates of 4.3 and 0.5% for astrovirus were observed in children with acute gastroenteritis and those without gastroenteritis, respectively. Among children with gastroenteritis no statistically significant difference was seen with regards to viral positivity rates in relation to gender and age. However, a higher incidence rate was observed for children from Brasília aged 36 months or more. Overall, astroviruses occurred predominantly from September to March in the two cities, suggesting a seasonal pattern for these viruses which coincides with the highest relative air humidity period. The results of this study highlight the importance of astrovirus as an etiologic agent of acute gastroenteritis in children of the Central West region of Brazil.

  17. Human astrovirus infection in a patient with new-onset celiac disease

    NARCIS (Netherlands)

    S.L. Smits (Saskia); M. van Leeuwen (Marije); A.A. Eijck (Annemiek); P.L.A. Fraaij (Pieter); J.C. Escher (Johanna); J.H. Simon (James); A.D.M.E. Osterhaus (Albert)

    2010-01-01

    textabstractMany diseases with unknown etiology may be caused by unidentified viruses. Sequence-independent amplification revealed a new astrovirus, similar to VA1, in a 4-year-old male diagnosed with celiac disease. This expands the geographic range of this virus to include Europe and may associate

  18. Astrovirus, reovirus and rotavirus concomitant infection causes decreased weight gain in broad-breasted white poults

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    Turkey astrovirus type-2 (TAstV-2), turkey rotavirus (TRotV) and turkey reovirus (TReoV) were evaluated for pathogenesis in 3 day-old turkey poults in all possible combinations of one, two or three viruses. Body-weights were recorded at 2, 4, 7, 10 and 14 days post inoculation (PI) and were decreas...

  19. Population-Based Incidence Rates of Diarrheal Disease Associated with Norovirus, Sapovirus, and Astrovirus in Kenya.

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    Kayoko Shioda

    Full Text Available Diarrheal diseases remain a major cause of mortality in Africa and worldwide. While the burden of rotavirus is well described, population-based rates of disease caused by norovirus, sapovirus, and astrovirus are lacking, particularly in developing countries.Data on diarrhea cases were collected through a population-based surveillance platform including healthcare encounters and household visits in Kenya. We analyzed data from June 2007 to October 2008 in Lwak, a rural site in western Kenya, and from October 2006 to February 2009 in Kibera, an urban slum. Stool specimens from diarrhea cases of all ages who visited study clinics were tested for norovirus, sapovirus, and astrovirus by RT-PCR.Of 334 stool specimens from Lwak and 524 from Kibera, 85 (25% and 159 (30% were positive for norovirus, 13 (4% and 31 (6% for sapovirus, and 28 (8% and 18 (3% for astrovirus, respectively. Among norovirus-positive specimens, genogroup II predominated in both sites, detected in 74 (87% in Lwak and 140 (88% in Kibera. The adjusted community incidence per 100,000 person-years was the highest for norovirus (Lwak: 9,635; Kibera: 4,116, followed by astrovirus (Lwak: 3,051; Kibera: 440 and sapovirus (Lwak: 1,445; Kibera: 879. For all viruses, the adjusted incidence was higher among children aged <5 years (norovirus: 22,225 in Lwak and 17,511 in Kibera; sapovirus: 5,556 in Lwak and 4,378 in Kibera; astrovirus: 11,113 in Lwak and 2,814 in Kibera compared to cases aged ≥5 years.Although limited by a lack of controls, this is the first study to estimate the outpatient and community incidence rates of norovirus, sapovirus, and astrovirus across the age spectrum in Kenya, suggesting a substantial disease burden imposed by these viruses. By applying adjusted rates, we estimate approximately 2.8-3.3 million, 0.45-0.54 million, and 0.77-0.95 million people become ill with norovirus, sapovirus, and astrovirus, respectively, every year in Kenya.

  20. A Review of the Strain Diversity and Pathogenesis of Chicken Astrovirus

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    Smyth, Victoria J.

    2017-01-01

    Although a relatively recently emerged virus, identified only in 2004 as a separate species of avian astrovirus, chicken astrovirus (CAstV) has been associated with poor growth of broiler flocks, enteritis and diarrhea and is a candidate pathogen in cases of runting stunting syndrome. More recently CAstV has been implicated in cases of two other diseases of broilers as the sole etiological agent, namely severe kidney disease of young broilers with visceral gout and the “White Chicks” hatchery disease. Examination of the strains of CAstV associated with the two latter diseases reveals they are closely related genetically. This review will discuss the pathogenesis of CAstV in relation to strain diversity and the effects of vertical versus horizontal transmission, virus load, co-infections and age of bird at infection, all factors that may impact upon disease severity. PMID:28208602

  1. Non-Human Primates Harbor Diverse Mammalian and Avian Astroviruses Including Those Associated with Human Infections.

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    Erik A Karlsson

    Full Text Available Astroviruses (AstVs are positive sense, single-stranded RNA viruses transmitted to a wide range of hosts via the fecal-oral route. The number of AstV-infected animal hosts has rapidly expanded in recent years with many more likely to be discovered because of the advances in viral surveillance and next generation sequencing. Yet no study to date has identified human AstV genotypes in animals, although diverse AstV genotypes similar to animal-origin viruses have been found in children with diarrhea and in one instance of encephalitis. Here we provide important new evidence that non-human primates (NHP can harbor a wide variety of mammalian and avian AstV genotypes, including those only associated with human infection. Serological analyses confirmed that >25% of the NHP tested had antibodies to human AstVs. Further, we identified a recombinant AstV with parental relationships to known human AstVs. Phylogenetic analysis suggests AstVs in NHP are on average evolutionarily much closer to AstVs from other animals than are AstVs from bats, a frequently proposed reservoir. Our studies not only demonstrate that human astroviruses can be detected in NHP but also suggest that NHP are unique in their ability to support diverse AstV genotypes, further challenging the paradigm that astrovirus infection is species-specific.

  2. The circulation of human astrovirus genotypes in the Central West Region of Brazil

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    Paula Andreia Silva

    2009-07-01

    Full Text Available Out of 1,588 faecal samples of children taken from three locations of the Central West Region of Brazil, 57 were positive for astroviruses (HAstVs using reverse transcription-polymerase chain reaction (RT-PCR. They were genotyped by nested RT-PCR and/or genomic sequencing. HAstV-1 (42.8%, HAstV-2 (23.2%, HAstV-3 (3.6%, HAstV-4 (14.3% and HAstVs -5, -6, -7 and -8 (1.8% each were detected. In Goiânia and Campo Grande, HAstV-1 was the most frequently detected genotype while in Brasília (DF it was HAstV-2. Shifts in the circulation of astrovirus genotypes were observed in DF and Campo Grande. All samples collected by rectal swabs were viral negative. The astrovirus genotypes were detected in all age groups and there was no correlation between genotype and age group.

  3. Prevalence of Rotavirus, Adenovirus, Norovirus, and Astrovirus Infections and Coinfections among Hospitalized Children in Northern France▿

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    Tran, Adissa; Talmud, Déborah; Lejeune, Benoît; Jovenin, Nicolas; Renois, Fanny; Payan, Christopher; Leveque, Nicolas; Andreoletti, Laurent

    2010-01-01

    From January to December 2007, 973 stool specimens were prospectively collected from children hospitalized for gastroenteritis signs or from neonates and premature cases who were born in two French hospital settings in the north of France. They were tested by rapid enzyme immunoassay (EIA) analyses for rotavirus and adenovirus and by two commercially available ELISA tests for the detection of norovirus and astrovirus. The overall rates of prevalence for rotavirus, norovirus, adenovirus, and astrovirus were 21, 13, 5, and 1.8%, respectively, and they did not significantly differ between the two hospital settings (P = 0.12). Mixed virus infections were detected in 32 (3.3%) of the 973 study children and were associated with norovirus in 21 (66%) infants, including 5 premature cases. From fall to spring, norovirus infections accounted for 52% of documented gastroenteritidis viral infections at a time when rotavirus was epidemic, resulting in mixed norovirus and rotavirus gastrointestinal tract infections. Of the 367 documented viral gastroenteritis cases, 15 (4.1%) were identified as nosocomial infections, 5 of which occurred in premature cases. These findings highlight the need to implement norovirus and astrovirus ELISA detection assays in association with rapid EIA rotavirus and adenovirus detection assays for the clinical diagnosis and the nosocomial prevention of gastroenteritis viral infections in pediatric departments. PMID:20305010

  4. Sewage surveillance reveals the presence of canine GVII norovirus and canine astrovirus in Uruguay.

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    Lizasoain, A; Tort, L F L; García, M; Gómez, M M; Leite, J P G; Miagostovich, M P; Cristina, J; Berois, M; Colina, R; Victoria, Matías

    2015-11-01

    Canine norovirus (NoV) and astrovirus (AstV) were studied in 20 domestic sewage samples collected in two cities in Uruguay. Four samples were characterized as canine AstV after phylogenetic analysis clustering with strains detected in Italy and Brazil in 2008 and 2012, respectively. One sample was characterized as canine NoV and clustered with a strain detected in Hong Kong and recently classified as GVII. This study shows the occurrence of a canine NoV GVII strain for the first time in the American continent and also warns about possible zoonotic infection, since canine strains were detected in domestic sewage.

  5. Establishment of Stably Transfected Cells Constitutively Expressing the Full-Length and Truncated Antigenic Proteins of Two Genetically Distinct Mink Astroviruses

    DEFF Research Database (Denmark)

    Bidokhti, Mehdi R. M.; Ullman, Karin; Jensen, Trine Hammer

    2013-01-01

    Astroviruses are becoming a growing concern in veterinary and public health. To date there are no registered vaccines against astrovirus-induced disease, mostly due to the difficulty to cultivate astroviruses to high titer for vaccine development using conventional techniques. As means to circumv......Astroviruses are becoming a growing concern in veterinary and public health. To date there are no registered vaccines against astrovirus-induced disease, mostly due to the difficulty to cultivate astroviruses to high titer for vaccine development using conventional techniques. As means...... to circumvent this drawback, we have developed stably transfected mink fetal cells and BHK21 cells constitutively expressing the full-length and truncated capsid proteins of two distinct genotypes of mink astrovirus. Protein expression in these stably transfected cells was demonstrated by strong signals...... shedding was observed in mink kits born from immunized females. The gene integration and protein expression were sustained through cell passage, showing that the used approach is robust and reliable for expression of functional capsid proteins for vaccine and diagnostic applications....

  6. Insectivorous bats carry host specific astroviruses and coronaviruses across different regions in Germany.

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    Fischer, Kerstin; Zeus, Veronika; Kwasnitschka, Linda; Kerth, Gerald; Haase, Martin; Groschup, Martin H; Balkema-Buschmann, Anne

    2016-01-01

    Recently several infectious agents with a zoonotic potential have been detected in different bat species. However, there is still a lack of knowledge on the transmission dynamics within and between bat species, as well as from bats to other mammals. To better understand these processes, it is important to compare the phylogenetic relationships between different agents to that of their respective hosts. In this study, we analysed more than 950 urine, faeces and oral swab samples collected from 653 bats from mainly four species (Myotis nattereri, Myotis bechsteinii, Myotis daubentonii, and Plecotus auritus) for the presence of coronavirus, paramyxovirus and astrovirus related nucleic acids located in three different regions of Germany. Using hemi-nested reverse transcriptase (RT)-PCR amplification of fragments within the highly conserved regions of the respective RNA dependent RNA polymerase (RdRp) genes, we detected astrovirus sequences at an overall detection rate of 25.8% of the analysed animals, with a maximum of 65% in local populations. The detection rates for coronaviruses and paramyxoviruses were distinctly lower, ranging between 1.4% and 3.1%. Interestingly, the sequence similarities in samples collected from the same bat species in different geographical areas were distinctly larger than the sequence similarities between samples from different species sampled at the same location. This indicates that host specificity may be more important than host ecology for the presence of certain viruses in bats.

  7. Structure of a Human Astrovirus Capsid-Antibody Complex and Mechanistic Insights into Virus Neutralization

    Energy Technology Data Exchange (ETDEWEB)

    Bogdanoff, Walter A.; Campos, Jocelyn; Perez, Edmundo I.; Yin, Lu; Alexander, David L.; DuBois, Rebecca M. (UCSC)

    2016-11-02

    ABSTRACT

    Human astroviruses (HAstVs) are a leading cause of viral diarrhea in young children, the immunocompromised, and the elderly. There are no vaccines or antiviral therapies against HAstV disease. Several lines of evidence point to the presence of protective antibodies in healthy adults as a mechanism governing protection against reinfection by HAstV. However, development of anti-HAstV therapies is hampered by the gap in knowledge of protective antibody epitopes on the HAstV capsid surface. Here, we report the structure of the HAstV capsid spike domain bound to the neutralizing monoclonal antibody PL-2. The antibody uses all six complementarity-determining regions to bind to a quaternary epitope on each side of the dimeric capsid spike. We provide evidence that the HAstV capsid spike is a receptor-binding domain and that the antibody neutralizes HAstV by blocking virus attachment to cells. We identify patches of conserved amino acids that overlap the antibody epitope and may comprise a receptor-binding site. Our studies provide a foundation for the development of therapies to prevent and treat HAstV diarrheal disease.

    IMPORTANCEHuman astroviruses (HAstVs) infect nearly every person in the world during childhood and cause diarrhea, vomiting, and fever. Despite the prevalence of this virus, little is known about how antibodies in healthy adults protect them against reinfection. Here, we determined the crystal structure of a complex of the HAstV capsid protein and a virus-neutralizing antibody. We show that the antibody binds to the outermost spike domain of the capsid, and we provide evidence that the antibody blocks virus attachment to human cells. Importantly, our findings suggest that a subunit-based vaccine focusing the immune system on the HAstV capsid spike domain could be effective in protecting children against HAstV disease.

  8. Astrovirus Infection in Hospitalized Infants with Severe Combined Immunodeficiency after Allogeneic Hematopoietic Stem Cell Transplantation

    Science.gov (United States)

    Berger, Christoph; Greiner, Oliver; Caduff, Rosmarie; Trkola, Alexandra; Bossart, Walter; Gerlach, Daniel; Schibler, Manuel; Cordey, Samuel; McKee, Thomas Alexander; Van Belle, Sandra; Kaiser, Laurent; Tapparel, Caroline

    2011-01-01

    Infants with severe primary combined immunodeficiency (SCID) and children post-allogeneic hematopoietic stem cell transplantation (HSCT) are extremely susceptible to unusual infections. The lack of generic tools to detect disease-causing viruses among more than 200 potential human viral pathogens represents a major challenge to clinicians and virologists. We investigated retrospectively the causes of a fatal disseminated viral infection with meningoencephalitis in an infant with gamma C-SCID and of chronic gastroenteritis in 2 other infants admitted for HSCT during the same time period. Analysis was undertaken by combining cell culture, electron microscopy and sequence-independent single primer amplification (SISPA) techniques. Caco-2 cells inoculated with fecal samples developed a cytopathic effect and non-enveloped viral particles in infected cells were detected by electron microscopy. SISPA led to the identification of astrovirus as the pathogen. Both sequencing of the capsid gene and the pattern of infection suggested nosocomial transmission from a chronically excreting index case to 2 other patients leading to fatal infection in 1 and to transient disease in the others. Virus-specific, real-time reverse transcription polymerase chain reaction was then performed on different stored samples to assess the extent of infection. Infection was associated with viremia in 2 cases and contributed to death in 1. At autopsy, viral RNA was detected in the brain and different other organs, while immunochemistry confirmed infection of gastrointestinal tissues. This report illustrates the usefulness of the combined use of classical virology procedures and modern molecular tools for the diagnosis of unexpected infections. It illustrates that astrovirus has the potential to cause severe disseminated lethal infection in highly immunocompromised pediatric patients. PMID:22096580

  9. Astrovirus infection in hospitalized infants with severe combined immunodeficiency after allogeneic hematopoietic stem cell transplantation.

    Directory of Open Access Journals (Sweden)

    Werner Wunderli

    Full Text Available Infants with severe primary combined immunodeficiency (SCID and children post-allogeneic hematopoietic stem cell transplantation (HSCT are extremely susceptible to unusual infections. The lack of generic tools to detect disease-causing viruses among more than 200 potential human viral pathogens represents a major challenge to clinicians and virologists. We investigated retrospectively the causes of a fatal disseminated viral infection with meningoencephalitis in an infant with gamma C-SCID and of chronic gastroenteritis in 2 other infants admitted for HSCT during the same time period. Analysis was undertaken by combining cell culture, electron microscopy and sequence-independent single primer amplification (SISPA techniques. Caco-2 cells inoculated with fecal samples developed a cytopathic effect and non-enveloped viral particles in infected cells were detected by electron microscopy. SISPA led to the identification of astrovirus as the pathogen. Both sequencing of the capsid gene and the pattern of infection suggested nosocomial transmission from a chronically excreting index case to 2 other patients leading to fatal infection in 1 and to transient disease in the others. Virus-specific, real-time reverse transcription polymerase chain reaction was then performed on different stored samples to assess the extent of infection. Infection was associated with viremia in 2 cases and contributed to death in 1. At autopsy, viral RNA was detected in the brain and different other organs, while immunochemistry confirmed infection of gastrointestinal tissues. This report illustrates the usefulness of the combined use of classical virology procedures and modern molecular tools for the diagnosis of unexpected infections. It illustrates that astrovirus has the potential to cause severe disseminated lethal infection in highly immunocompromised pediatric patients.

  10. Molecular epidemiology and phylogenetic analysis of diverse bovine astroviruses associated with diarrhea in cattle and water buffalo calves in China.

    Science.gov (United States)

    Alfred, Niyokwishimira; Liu, Huan; Li, Mu Lan; Hong, Shao Feng; Tang, Hai Bo; Wei, Zu Zhang; Chen, Ying; Li, Fa Kai; Zhong, Yi Zhi; Huang, Wei Jian

    2015-06-01

    Astroviruses are the principal causative agents of gastroenteritis in humans and have been associated with diarrhea in other mammals as well as birds. However, astroviral infection of animals had been poorly studied. In the present study, 211 rectal swabs collected from cattle and water buffalo calves with mild to severe diarrhea were tested for bovine astrovirus (BAstV) by RT-PCR. Results: 92/211 (43.6%) samples were positive for BAstV, at a rate of 46.10% (71/154) in cattle and 36.84% (21/57) in water buffalo. Phylogenetic analysis based on the partial and full-length of 25 ORF2 amino acid sequences obtained in this study classified the Guangxi BAstVs isolates into five subgroups under the genus of Mamastrovirus, genotype MAstV33, which suggested that the water buffalo was a new host of this genogroup that previously included only cattle and roe deer. Despite the origin of the host, the Guangxi BAstV isolates were closely related to the BAstV Hong Kong isolates (B18/HK and B76-2/HK), but highly divergent from the BAstV NeuroS1 isolate previously associated with neurologic disease in cattle in the U.S.A. Nucleotide sequence-based characterization of the ORF1b/ORF2 junction and corresponding overlapping regions showed distinctive properties, which may be common to BAstVs. Our results suggested that cattle and water buffalo are prone to infection of closely related astroviruses, which probably evolved from the same ancestor. The current study described astroviruses in water buffalo for the first time and is thus far among the largest epidemiological investigations of BAstV infection in cattle conducted in China.

  11. Identification of an astrovirus commonly infecting laboratory mice in the US and Japan.

    Science.gov (United States)

    Ng, Terry Fei Fan; Kondov, Nikola O; Hayashimoto, Nobuhito; Uchida, Ritsuki; Cha, Yunhee; Beyer, Ashley I; Wong, Walt; Pesavento, Patricia A; Suemizu, Hiroshi; Muench, Marcus O; Delwart, Eric

    2013-01-01

    Mice (Mus musculus) are the most commonly used laboratory animals. Viral metagenomics on tissues of immunodeficient mice revealed sequences of a novel mammalian astrovirus. Using PCR, we screened mice from 4 breeders, 4 pharmaceutical companies, 14 research institutes and 30 universities in the US and Japan. Mice from one US breeder tested positive while none from Japanese breeders were positive for MuAstV. Mice in over half of the universities (19/30), institutes (7/14) and pharmaceutical animal facilities (2/4) investigated revealed the presence of MuAstV. Nine mice strains tested positive including both immunodeficient strains (NSG, NOD-SCID, NSG-3GS, C57BL6-Timp-3 (-/-), and uPA-NOG) and immunocompetent strains (B6J, ICR, Bash2, BALB/c). Our data indicates that MuAstV has a wide geographical, institutional and host strain distribution. Comparison of the MuAstV RdRp sequences showed numerous mutations indicating ongoing viral divergence in different facilities. This study demonstrates the need for metagenomic screening of laboratory animals to identify adventitious infections that may affect experimental outcomes.

  12. Identification of an astrovirus commonly infecting laboratory mice in the US and Japan.

    Directory of Open Access Journals (Sweden)

    Terry Fei Fan Ng

    Full Text Available Mice (Mus musculus are the most commonly used laboratory animals. Viral metagenomics on tissues of immunodeficient mice revealed sequences of a novel mammalian astrovirus. Using PCR, we screened mice from 4 breeders, 4 pharmaceutical companies, 14 research institutes and 30 universities in the US and Japan. Mice from one US breeder tested positive while none from Japanese breeders were positive for MuAstV. Mice in over half of the universities (19/30, institutes (7/14 and pharmaceutical animal facilities (2/4 investigated revealed the presence of MuAstV. Nine mice strains tested positive including both immunodeficient strains (NSG, NOD-SCID, NSG-3GS, C57BL6-Timp-3 (-/-, and uPA-NOG and immunocompetent strains (B6J, ICR, Bash2, BALB/c. Our data indicates that MuAstV has a wide geographical, institutional and host strain distribution. Comparison of the MuAstV RdRp sequences showed numerous mutations indicating ongoing viral divergence in different facilities. This study demonstrates the need for metagenomic screening of laboratory animals to identify adventitious infections that may affect experimental outcomes.

  13. Serological studies confirm the novel astrovirus HMOAstV-C as a highly prevalent human infectious agent.

    Directory of Open Access Journals (Sweden)

    Peter D Burbelo

    Full Text Available Molecular identification of a microbe is the first step in determining its prevalence of infection and pathogenic potential. Detection of specific adaptive immune responses can provide insights into whether a microbe is a human infectious agent and its epidemiology. Here we characterized human anti-IgG antibody responses by luciferase immunoprecipitation systems (LIPS against two protein fragments derived from the capsid protein of the novel HMOAstV-C astrovirus. While antibodies to the N-terminal fragment were not informative, the C-terminal capsid fragment of HMOAstV-C showed a high frequency of immunoreactivity with serum from healthy blood donors. In contrast, a similar C-terminal capsid fragment from the related HMOAstV-A astrovirus failed to show immunoreactivity. Detailed analysis of adult serum from the United Sates using a standardized threshold demonstrated HMOAstV-C seropositivity in approximately 65% of the samples. Evaluation of serum samples from different pediatric age groups revealed that the prevalence of antibodies in 6-12 month, 1-2 year, 2-5 year and 5-10 year olds was 20%, 23%, 32% and 36%, respectively, indicating rising seroprevalence with age. Additionally, 50% (11/22 of the 0-6 month old children showed anti-HMOAstV-C antibody responses, likely reflecting maternal antibodies. Together these results document human humoral responses to HMOAstV-C and validate LIPS as a facile and effective approach for identifying humoral responses to novel infectious agents.

  14. Frequency and Pathological Phenotype of Bovine Astrovirus CH13/NeuroS1 Infection in Neurologically-Diseased Cattle: Towards Assessment of Causality

    Directory of Open Access Journals (Sweden)

    Senija Selimovic-Hamza

    2017-01-01

    Full Text Available Next-generation sequencing (NGS has opened up the possibility of detecting new viruses in unresolved diseases. Recently, astrovirus brain infections have been identified in neurologically diseased humans and animals by NGS, among them bovine astrovirus (BoAstV CH13/NeuroS1, which has been found in brain tissues of cattle with non-suppurative encephalitis. Only a few studies are available on neurotropic astroviruses and a causal relationship between BoAstV CH13/NeuroS1 infections and neurological disease has been postulated, but remains unproven. Aiming at making a step forward towards assessing the causality, we collected brain samples of 97 cases of cattle diagnosed with unresolved non-suppurative encephalitis, and analyzed them by in situ hybridization and immunohistochemistry, to determine the frequency and neuropathological distribution of the BoAstV CH13/NeuroS1 and its topographical correlation to the pathology. We detected BoAstV CH13/NeuroS1 RNA or proteins in neurons throughout all parts of the central nervous system (CNS in 34% of all cases, but none were detected in cattle of the control group. In general, brain lesions had a high correlation with the presence of the virus. These findings show that a substantial proportion of cattle with non-suppurative encephalitis are infected with BoAstV CH13/NeuroS1 and further substantiate the causal relationship between neurological disease and astrovirus infections.

  15. Frequency and Pathological Phenotype of Bovine Astrovirus CH13/NeuroS1 Infection in Neurologically-Diseased Cattle: Towards Assessment of Causality

    Science.gov (United States)

    Selimovic-Hamza, Senija; Boujon, Céline L.; Hilbe, Monika; Oevermann, Anna; Seuberlich, Torsten

    2017-01-01

    Next-generation sequencing (NGS) has opened up the possibility of detecting new viruses in unresolved diseases. Recently, astrovirus brain infections have been identified in neurologically diseased humans and animals by NGS, among them bovine astrovirus (BoAstV) CH13/NeuroS1, which has been found in brain tissues of cattle with non-suppurative encephalitis. Only a few studies are available on neurotropic astroviruses and a causal relationship between BoAstV CH13/NeuroS1 infections and neurological disease has been postulated, but remains unproven. Aiming at making a step forward towards assessing the causality, we collected brain samples of 97 cases of cattle diagnosed with unresolved non-suppurative encephalitis, and analyzed them by in situ hybridization and immunohistochemistry, to determine the frequency and neuropathological distribution of the BoAstV CH13/NeuroS1 and its topographical correlation to the pathology. We detected BoAstV CH13/NeuroS1 RNA or proteins in neurons throughout all parts of the central nervous system (CNS) in 34% of all cases, but none were detected in cattle of the control group. In general, brain lesions had a high correlation with the presence of the virus. These findings show that a substantial proportion of cattle with non-suppurative encephalitis are infected with BoAstV CH13/NeuroS1 and further substantiate the causal relationship between neurological disease and astrovirus infections. PMID:28106800

  16. An outbreak of gastroenteritis associated with astrovirus serotype 1 in a day care center, in Rio de Janeiro, Brazil

    Directory of Open Access Journals (Sweden)

    AMV Silva

    2001-11-01

    Full Text Available Between June 4th and June 20th1996 rotavirus, adenovirus, and astrovirus (HAstrV were investigated in fecal samples from 27 children under three years old with acute diarrhea, attending the Bertha Lutz day care center, in Rio de Janeiro. All fecal samples were analyzed by polyacrylamide gel electrophoresis (PAGE, reverse transcriptase polymerase chain reaction (RT-PCR, enzyme immunoassays (EIA, and electron microscopy (EM. Nine of them (33% showed positive results for HAstrV by at least one of the employed methodologies. Eight were positive by RT-PCR and EIA, and six by EM. All positive samples were inoculated onto HT-29 (human colon adenocarcinoma cultured cells for HAstrV isolation and seven were positive after three passages. The sequencing analysis of eight RT-PCR products (449 bp from gene that codifies VP2 protein, showed a total nucleotide identity among them and 98% with HAstrV-1 (strain Oxford type 1. This is the first report of a gastroenteritis outbreak associated with HAstrv-1 in a day care center in Rio de Janeiro and it reinforces the importance of this virus in association with infantile acute gastroenteritis.

  17. Astrovirus (HAstV) como agente causal de diarrea en niños Colombianos: Siete años de estudio

    OpenAIRE

    Gutiérrez, María Fernanda; Matiz, Adriana; Alvarado, Mónica; Ulloa, Juan Carlos

    2005-01-01

    Con el objeto de determinar el comportamiento de los Astrovirus como agentes causantes de diarrea en algunas regiones colombianas, se colectaron 1147 muestras de diarrea de niños menores de 5 años que acudieron a los servicios de consulta externa de cuatro centros de salud de regiones de Bogotá, Facatativa, Cartagena y Quibdo durante el periodo comprendido entre 1996 y el año 2002. La determinación de la presencia viral se realizó por medio de pruebas de ELISA encontrando una prevalencia glob...

  18. Diversity of human astrovirus genotypes circulating in children with acute gastroenteritis in Thailand during 2000-2011.

    Science.gov (United States)

    Malasao, Rungnapa; Khamrin, Pattara; Chaimongkol, Natthawan; Ushijima, Hiroshi; Maneekarn, Niwat

    2012-11-01

    Human astrovirus (HAstV) is one of the causative agents of acute gastroenteritis in children worldwide. The objective of this study was to elucidate the molecular epidemiology and genotypic diversity of HAstV circulating in pediatric patients admitted to hospital with diarrhea in Thailand during the year 2000-2011, except for 2004, 2006, and 2009. A total of 1,022 fecal specimens were tested for HAstV by reverse transcription polymerase chain reaction (RT-PCR). HAstV was detected at 1.4% (14 of 1,022). All HAstV strains detected in this study were characterized further by nucleotide sequencing and phylogenetic analysis. Analysis of 348 bp partial capsid nucleotide sequences revealed that HAstV strains detected were HAstV-1 (1a, 1b, and 1d) (8 strains), HAstV-2 (2c) (3 strains), HAstV-3 (1 strain), and HAstV-5 (2 strains). HAstV-1, the most predominant genotype was detected initially in 2002 and circulated continuously up to 2011. HAstV-2 was detected in year 2001, and 2007 and grouped into a 2c lineage. HAstV-3 was found only in 2000 and HAstV-5 was found in the year 2001. The findings indicate that a wide variety of HAstV strains continue to circulate in children admitted to hospital with acute gastroenteritis in Thailand over a decade. The data provide an epidemiological overview of HAstV infection and HAstV genotype distribution in Thailand.

  19. PTB Binds to the 3’ Untranslated Region of the Human Astrovirus Type 8: A Possible Role in Viral Replication

    Science.gov (United States)

    Espinosa-Hernández, Wendy; Velez-Uriza, Dora; Valdés, Jesús; Vélez-Del Valle, Cristina; Salas-Benito, Juan; Martínez-Contreras, Rebeca; García-Espítia, Matilde; Salas-Benito, Mariana; Vega-Almeida, Tania; De Nova-Ocampo, Mónica

    2014-01-01

    The 3′ untranslated region (3′UTR) of human astroviruses (HAstV) consists of two hairpin structures (helix I and II) joined by a linker harboring a conserved PTB/hnRNP1 binding site. The identification and characterization of cellular proteins that interact with the 3′UTR of HAstV-8 virus will help to uncover cellular requirements for viral functions. To this end, mobility shift assays and UV cross-linking were performed with uninfected and HAstV-8-infected cell extracts and HAstV-8 3′UTR probes. Two RNA-protein complexes (CI and CII) were recruited into the 3′UTR. Complex CII formation was compromised with cold homologous RNA, and seven proteins of 35, 40, 45, 50, 52, 57/60 and 75 kDa were cross-linked to the 3′UTR. Supermobility shift assays indicated that PTB/hnRNP1 is part of this complex, and 3′UTR-crosslinked PTB/hnRNP1 was immunoprecipitated from HAstV-8 infected cell-membrane extracts. Also, immunofluorescence analyses revealed that PTB/hnRNP1 is distributed in the nucleus and cytoplasm of uninfected cells, but it is mainly localized perinuclearly in the cytoplasm of HAstV-8 infected cells. Furthermore, the minimal 3′UTR sequences recognized by recombinant PTB are those conforming helix I, and an intact PTB/hnRNP1-binding site. Finally, small interfering RNA-mediated PTB/hnRNP1 silencing reduced synthesis viral genome and virus yield in CaCo2 cells, suggesting that PTB/hnRNP1 is required for HAstV replication. In conclusion, PTB/hnRNP1 binds to the 3′UTR HAstV-8 and is required or participates in viral replication. PMID:25406089

  20. The prevalence of norovirus, astrovirus and adenovirus infections among hospitalised children with acute gastroenteritis in Porto Velho, state of Rondônia, western Brazilian Amazon

    Directory of Open Access Journals (Sweden)

    Maria Sandra Costa Amaral

    2015-04-01

    Full Text Available Although viruses are well-established causes of acute gastroenteritis, few data on the circulation of these pathogens in Porto Velho, state of Rondônia, Brazil, are available. Thus, faecal samples from hospitalised diarrhoeic children, under six years of age, were collected and tested for the presence of norovirus (NoV, adenovirus (AdV and astrovirus (AstV from February 2010-February 2012. Specimens were screened by reverse-transcription polymerase chain reaction and viruses were found in 10.7% (63/591 of the cases. NoV, AdV and AstV were detected in 7.8%, 2% and 0.8% of the samples, respectively. NoV infection was observed at all ages and was most prevalent in zero-18-month-old children (84.7%; p = 0.002. A higher incidence of NoV was detected from February-April 2010, when it was found in 52.2% of the cases. Co-infections involving these viruses, rotavirus and enteropathogenic bacteria were detected in 44.4% (28/63 of the children with viral diarrhoea. Nosocomial infections were demonstrated in 28.6% (18/63 of the cases in which viruses were detected. The present paper reports, for the first time, the circulation of NoV and AstV among the paediatric population of Porto Velho and it contributes to our understanding of the roles of these pathogens in gastrointestinal infections.

  1. Epidemiological aspects of astrovirus and coronavirus in poults in the South Eastern Region of Brazil Aspectos epidemiológicos associados à prevalência de Astrovirus e Coronavirus em lotes de perus da Região Sudeste do Brasil

    Directory of Open Access Journals (Sweden)

    S.E.L. da Silva

    2009-06-01

    Full Text Available A survey of Turkey Coronavirus (TCoV and Astrovirus (TAstV-2 prevalence was carried out from February to December during 2006 year in semiarid region of Brazil, from a turkey producer area, localized in South Eastern of Brazil. To asses the risk factor related to clinical material, climatic condition and type of RT-PCR applied, cloacal swabs (CS, faeces, sera, bursa of Fabricius (BF, thymus (TH and spleen (SP and ileum-caeca region were collected from 30-day-old poults suffering of enteritis episode characterized as poult enteritis mortality syndrome (PEMS. The PEMS clinical features were characterized by watery to foamy faeces, light brown-yellow in colour and low mortality rate. Meteorological data (rainfall and relative humidity observed during along the study presented monthly average temperature ranging from 39.3 and 31.2ºC, precipitation in rainy season from 40 to 270.3 mm/month, and no rain during dry season. Simplex RT-PCR gave odds ratio (OR values suggesting that ileum-caeca region is at higher chance (OR=1.9; p=0.9741 to have both viral RNA than faeces (OR=1.5; p=0.7319. However, multiplex RT-PCR showed 3.98 (p=0.89982 more chance to give positive results in faeces than CS at dry season. The major risk factors seem to be low rate of humidity and high temperatures at winter, probably responsible for spread, easily, the TCoV and TAstv-2 among the flocks. The positive results of both virus suggested that they can play an important role in enteric disorders, associated to low humidity and high temperatures frequently found in tropical countries.O presente estudo foi conduzido para avaliar a prevalência do Coronavirus dos perus (TCoV e Astrovirus tipo 2 (TAstV-2 entre os meses de Fevereiro a Dezembro de 2006, em uma região produtora localizada no semi-árido a Sudeste do Brasil. Os principais fatores de risco associado a prevalência foram material clínico analisado, condições climáticas e tipo de técnica molecular empregada. Os

  2. Identification and characterization of deer astroviruses

    DEFF Research Database (Denmark)

    Smits, Saskia L.; van Leeuwen, Marije; Kuiken, Thijs;

    2010-01-01

    The threat of emerging infectious viruses in humans requires a more effective approach regarding virus surveillance. A thorough understanding of virus diversity in wildlife provides epidemiological baseline information about pathogens and may lead to the identification of newly emerging pathogens...

  3. Gastroenteritis Attributable to 16 Enteropathogens in Children Attending Day Care Significant Effects of Rotavirus, Norovirus, Astrovirus, Cryptosporidium and Giardia

    NARCIS (Netherlands)

    Enserink, Remko; van den Wijngaard, Cees; Bruijning-Verhagen, Patricia; van Asten, Liselotte; Mughini-Gras, Lapo; Duizer, Erwin; Kortbeek, Titia; Scholts, Rianne; Nagelkerke, Nico; Smit, Henriette A.; Kooistra-Smid, Mirjam; van Pelt, Wilfrid

    2015-01-01

    Background: Children attending day care experience substantial gastrointestinal morbidity due to circulating seasonal enteropathogens in the day-care environment. The lack of a distinct clinical presentation of gastroenteritis (GE) in these children, in combination with the high diversity of enterop

  4. Molecular epidemiology of enteric viruses and genotyping of rotavirus A, adenovirus and astrovirus among children under 5 years old in Gabon

    Directory of Open Access Journals (Sweden)

    Sonia Etenna Lekana-Douki

    2015-05-01

    Conclusions: These findings improve our knowledge of circulating enteric viruses in Gabon. The emergence of unusual G6P[6] strain of rotavirus A, predominant, suggested a particular epidemiological surveillance of circulating rotavirus strains during the introduction of vaccination in Gabon.

  5. AcEST: BP917675 [AcEST

    Lifescience Database Archive (English)

    Full Text Available SD_TASV2 Capsid polyprotein OS=Turkey astrovirus 2 ... 30 4.2 sp|P09001|RM03_HUMAN 39S ribosomal protein L3,...RINELRRLKRKKEYH------NDEYKDDEIYLDNILKG 315 >sp|Q9Q3G5|CAPSD_TASV2 Capsid polyprotein OS=Turkey astrovirus 2

  6. Year-round screening of noncommercial and commercial oysters for the presence of human pathogenic viruses

    NARCIS (Netherlands)

    Lodder-Verschoor, F.; Husman, A.M.R.; Berg, van den H.H.J.L.; Stein, A.; Pelt-Heerschap, van H.M.L.; Poel, van der W.H.M.

    2005-01-01

    Consumption of virus-contaminated shellfish has caused numerous outbreaks of gastroenteritis and hepatitis worldwide. In The Netherlands, oysters are cultured and imported both for consumption and export; therefore, the presence of noroviruses, rotaviruses, astroviruses, hepatitis A viruses, and ent

  7. Complete genome sequence and integrated protein localization and interaction map for alfalfa dwarf virus, which combines properties of both cytoplasmic and nuclear plant rhabdoviruses

    Energy Technology Data Exchange (ETDEWEB)

    Bejerman, Nicolás, E-mail: n.bejerman@uq.edu.au [Instituto de Patología Vegetal (IPAVE), Centro de Investigaciones Agropecuarias (CIAP), Instituto Nacional de Tecnología Agropecuaria INTA, Camino a 60 Cuadras k 5,5, Córdoba X5020ICA (Argentina); Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, St Lucia, QLD 4072 (Australia); Giolitti, Fabián; Breuil, Soledad de; Trucco, Verónica; Nome, Claudia; Lenardon, Sergio [Instituto de Patología Vegetal (IPAVE), Centro de Investigaciones Agropecuarias (CIAP), Instituto Nacional de Tecnología Agropecuaria INTA, Camino a 60 Cuadras k 5,5, Córdoba X5020ICA (Argentina); Dietzgen, Ralf G. [Queensland Alliance for Agriculture and Food Innovation, The University of Queensland, St Lucia, QLD 4072 (Australia)

    2015-09-15

    Summary: We have determined the full-length 14,491-nucleotide genome sequence of a new plant rhabdovirus, alfalfa dwarf virus (ADV). Seven open reading frames (ORFs) were identified in the antigenomic orientation of the negative-sense, single-stranded viral RNA, in the order 3′-N-P-P3-M-G-P6-L-5′. The ORFs are separated by conserved intergenic regions and the genome coding region is flanked by complementary 3′ leader and 5′ trailer sequences. Phylogenetic analysis of the nucleoprotein amino acid sequence indicated that this alfalfa-infecting rhabdovirus is related to viruses in the genus Cytorhabdovirus. When transiently expressed as GFP fusions in Nicotiana benthamiana leaves, most ADV proteins accumulated in the cell periphery, but unexpectedly P protein was localized exclusively in the nucleus. ADV P protein was shown to have a homotypic, and heterotypic nuclear interactions with N, P3 and M proteins by bimolecular fluorescence complementation. ADV appears unique in that it combines properties of both cytoplasmic and nuclear plant rhabdoviruses. - Highlights: • The complete genome of alfalfa dwarf virus is obtained. • An integrated localization and interaction map for ADV is determined. • ADV has a genome sequence similarity and evolutionary links with cytorhabdoviruses. • ADV protein localization and interaction data show an association with the nucleus. • ADV combines properties of both cytoplasmic and nuclear plant rhabdoviruses.

  8. Characterization of an outbreak of astroviral diarrhea in a group of cheetahs (Acinonyx jubatus).

    Science.gov (United States)

    Atkins, Adrienne; Wellehan, James F X; Childress, April L; Archer, Linda L; Fraser, William A; Citino, Scott B

    2009-04-14

    A Mamastrovirus was identified in an outbreak of diarrhea in cheetahs (Acinonyx jubatus). Five young adult and two adult cheetahs presented with lethargy, anorexia, watery diarrhea and regurgitation over an 11-day period. Fecal samples were submitted for electron microscopy and culture. Electron microscopy results revealed particles morphologically consistent with an astrovirus, and no other viral pathogens or significant bacterial pathogens were identified. The astrovirus was confirmed and sequenced using consensus astroviral PCR, resulting in a 367 base pair partial RNA-dependent-RNA polymerase (RdRp) product and a 628 base pair partial capsid product. Bayesian and maximum likelihood phylogenetic analyses were performed on both the RdRp and the capsid protein segments. All animals were monitored and treated with bismuth subsalicylate tablets (524mg PO BID for 5 days), and recovered without additional intervention. This is the first report we are aware of documenting an astrovirus outbreak in cheetah.

  9. 海南省虫媒病毒调查研究状况%Review on arbovirus studies in Hainan province, China

    Institute of Scientific and Technical Information of China (English)

    郑雅匀; 王志玉; 梁国栋

    2011-01-01

    North to Torrid Zone, Hainan province has favorable natural conditions to arthropod reproduction and the spread of arboviruses, as evidenced by several outbreaks of dengue fever and endemic Japanese encephalitis (JE). Arboviruses, including four alphaviruses (Chikungunya virus, Mayaro virus, Getah virus, Ross River virus), two flaviviruses (JE virus and Dengue virus) and one seadornavirus, have been isolated from various vectors and host animals in this province. This article reviews current studies on arboviruses and arboviral infections to humans and livestocks, in order to inform prevention and control both locally and nationwide.%海南省地处热带北缘,自然条件适合节肢动物繁殖及虫媒病毒传播,曾发生过多次登革热大流行,也是流行性乙型脑炎(乙脑)地方性流行区.以往从海南省多种媒介和宿主动物中分离到4种甲病毒(基孔肯雅、马雅罗、盖塔和罗斯河病毒)、2种黄病毒(乙脑和登革热病毒)和1种Seadornavirus.现对海南省新分离虫媒病毒及其人畜感染状况进行回顾整理,为当地以及我国虫媒病毒病的预防控制提供依据.

  10. Determination and analysis of the full-length chicken parvovirus genome.

    Science.gov (United States)

    Viral enteric disease in poultry is an ongoing problem in many parts of the world. Many enteric viruses have been identified in turkeys and chickens, including avian astroviruses, rotaviruses, reoviruses, and coronaviruses. Through the application of a molecular screening method targeting particle-a...

  11. Partial Genome Sequence Analysis of Parvoviruses Associated with Enteric Disease in Poultry

    Science.gov (United States)

    Poult Enteritis Mortality Syndrome (PEMS) of turkeys and Runting-Stunting Syndrome (RSS) of chickens are significant viral enteric diseases of poultry. Although a number of different viruses, including avian reoviruses, rotaviruses, astroviruses and coronaviruses, have been isolated from the intesti...

  12. A Systematic Approach to Elucidate Causes of Gastroenteritis Outbreaks of Suspected Viral Etiology

    NARCIS (Netherlands)

    S. Svraka-Latifovic (Sanela)

    2011-01-01

    textabstractThe main objective of this thesis was to investigate the etiology of outbreaks of viral gastroenteritis that remained without diagnosis after testing for common viral pathogens causing gastroenteritis, e.g. noroviruses, rotaviruses, sapoviruses, adenoviruses, and astroviruses. No causati

  13. Molecular Epidemiology of Viral Gastroenteritis in Hajj pilgrimage

    KAUST Repository

    Padron Regalado, Eriko

    2014-05-01

    Hajj is the annual gathering of Islam practitioners in Mecca, Saudi Arabia. During the event, gastrointestinal infections are usually experienced and outbreaks have always been a concern; nevertheless, a deep and integrative study of the etiological agents has never been carried out. Here, I describe for the first time the epidemiology of pathogenic enteric viruses during Hajj 2011, 2012 and 2013. The focus of this study was the common enteric viruses Astrovirus, Norovirus, Rotavirus and Adenovirus. An enzyme Immunoassay established their presence in 14.9%, 15.0% and 6.6% of the reported cases of acute diarrhea for 2011, 2012 and 2013, respectively. For the three years of study, Astrovirus accounted for the majority of the viral infections. To our knowledge, this is the first time an epidemiological study depicts Astrovirus as the main viral agent of gastroenteritis in a mass gathering event. Hajj is rich in strains of Astrovirus, Norovirus and Rotavirus. A first screening by RT-PCR resulted in ten different genotypes. Strains HAstV 2, HAstV 1 and HAstV 5 were identified for Astrovirus. GI.6, GII.3, GII.4 and GII.1 were described for Norovirus and G1P[8], G4P[8] and G3P[8] were found for Rotavirus. The majority of the Astrovirus isolates could not be genotyped suggesting the presence of a new variant(s). Cases like this encourage the use of metagenomics (and nextgeneration sequencing) as a state-of-the-art technology in clinical diagnosis. A sample containing Adenovirus particles is being used to standardize a process for detection directly from stool samples and results will be obtained in the near future. The overall findings of the present study support the concept of Hajj as a unique mass gathering event that potentiates the transmission of infectious diseases. The finding of Norovirus GII.4 Sydney, a variant originated from Australia, suggests that Hajj is a receptor of infectious diseases worldwide. This work is part of the Hajj project, a collaborative

  14. Viral gastrointestinal syndrome in our environment

    Directory of Open Access Journals (Sweden)

    Patić A.

    2014-01-01

    Full Text Available Viral gastrointestinal syndrome is a cause of morbidity and death worldwide. Infection is spread through contact with an infected person, as well as through contaminated food and water. A lethal outcome is possible in infants and young children due to dehydration and electrolyte imbalance. The study included 141 patients with gastroenteritis from Vojvodina. Real-Time PCR method in stool samples was used to determine the presence of rota-, noro-, and astrovirus nucleic acid. Out of 141 patients with gastroenteritis, 60.3% were confirmed to have one of the three viruses. Rotavirus was significantly more common in children up to 3 years of age (43.3%. Norovirus was more frequently detected in patients older than 20 (50%. These infections started in collectives. Astrovirus was detected in four patients (2.8%. The results confirm the necessity to implement PCR in routine diagnostics for the proper treatment of patients.

  15. Viral causes of diarrhea.

    Science.gov (United States)

    Goodgame, R W

    2001-09-01

    Viruses are important causes of diarrhea. In healthy adults, the main clinical manifestation is acute, self-limited gastroenteritis. Advances in molecular diagnostics have shown that epidemics of acute gastroenteritis most frequently are due to caliciviruses spread through contaminated food or through person-to-person contact. Application of similar technology is needed to make a definitive statement about the role of such candidate viruses as rotavirus, astrovirus, and adenovirus as the cause of nonepidemic acute gastroenteritis in adults. Rarely a previously healthy adult gets acute CMV colitis. CMV and EBV mainly cause diarrhea in immunocompromised patients, however. Advances in prophylaxis and treatment have reduced the frequency and severity of these diseases. Acute infantile gastroenteritis is caused by rotavirus, calcivirus, astrovirus, and adenovirus. These viral diseases of the gut are seen by the physician as routine and rare clinical problems.

  16. High prevalence of turkey parvovirus in turkey flocks from Hungary experiencing enteric disease syndromes.

    Science.gov (United States)

    Palade, Elena Alina; Demeter, Zoltán; Hornyák, Akos; Nemes, Csaba; Kisary, János; Rusvai, Miklós

    2011-09-01

    Samples collected in 2008 and 2009, from 49 turkey flocks of 6 to 43 days in age and presenting clinical signs of enteric disease and high mortality, were tested by polymerase chain reaction and reverse transcription-polymerase chain reaction for the presence of viruses currently associated with enteric disease (ED) syndromes: astrovirus, reovirus, rotavirus, coronavirus, adenovirus, and parvovirus. Turkey astroviruses were found in 83.67% of the cases and turkey astrovirus 2 (TAst-2) in 26.53%. The investigations directly demonstrated the high prevalence of turkey parvovirus (TuPV) in 23 flocks (46.9%) experiencing signs of ED, making this pathogen the second most identified after astroviruses. Phylogenetic analysis on a 527 base pair-long region from the NS1 gene revealed two main clusters, a chicken parvovirus (ChPV) and a TuPV group, but also the presence of a divergent branch of tentatively named "TuPV-like ChPV" strains. The 23 Hungarian TuPV strains were separately positioned in two groups from the American origin sequences in the TuPV cluster. An Avail-based restriction fragment length polymorphism assay has also been developed for the quick differentiation of TuPV, ChPV, and divergent TuPV-like ChPV strains. As most detected enteric viruses have been directly demonstrated in healthy turkey flocks as well, the epidemiology of this disease complex remains unclear, suggesting that a certain combination of pathogens, environmental factors, or both are necessary for the development of clinical signs.

  17. THE ETIOLOGIC SPECTRUM OF PATHOGENS OF VIRAL GASTROENTERITIS IN CHILDREN FROM BAKU

    OpenAIRE

    N. N. Aliev; S. N. Musaev; N. A. Azizova; L. I. Rustamova; Z. M. Kulieva

    2016-01-01

    The article presents the results of research on the study of etiology, the logical structure of viral diarrhea in Baku (Azerbaijan) in 2015. It was found that more than half (62.6%), gastroenteritis in children of viral etiology, of which the leading role as an etiological factor, have a company — and adenoviruses, among infants astroviruses. But-roviral gastroenteritis and enterovirus takes only insignificant-tive percentage of cases. There were no significant differences in the proportion o...

  18. Prevalence and Genetic Diversity of Enteric Viruses in Children with Diarrhea in Ouagadougou, Burkina Faso.

    Directory of Open Access Journals (Sweden)

    Nafissatou Ouédraogo

    Full Text Available Enteric viruses are a major cause of diarrhea in children, especially those under five years old. Identifying the viral agents is critical to the development of effective preventive measures. This study aimed to determine the prevalence and genetic diversity of common enteric viruses in children under five years old in Burkina Faso. Stool samples from children with (n = 263 and without (n = 50 diarrhea disorders were collected in Ouagadougou, Burkina Faso from November 2011 to September 2012. Rotavirus, norovirus, sapovirus, astrovirus, adenovirus and Aichivirus A were detected using real-time or end-point (RT-PCR. Rotavirus strains were G and P genotyped by multiplex RT-PCR and other viral strains were characterized by sequencing of viral subgenomic segements. At least one viral agent was detected in 85.6% and 72% of the symptomatic and asymptomatic patients, respectively. Rotavirus (63.5%, adenovirus (31.2% and genogroup II norovirus (18.2% were the most prevalent viruses in symptomatic patients, but only rotavirus and genogroup II norovirus were significantly associated with diarrhea (OR: 7.9, 95%CI: 3.7-17; OR: 3.5, 95%CI: 1-11.7, respectively. Sapovirus (10.3%, astrovirus (4.9%, genogroup I norovirus (2.7% and Aichivirus A (0.8% were less prevalent. The predominant genotype of rotavirus was G9P[8] (36.5%, and the predominant norovirus strain was GII.4 variant 2012 (71.4%. Among sapovirus, the genogroup II (87.5% predominated. Astrovirus type 1 (41.7% was the most frequent astrovirus identified. Aichivirus A belonged to the three genotypes (A, B and C. Enteric adenoviruses type 40 and 41 were identified in 10.2% and 5.1% respectively. Several cases of co-infections were detected. The results highlight the high prevalence and the high diversity of enteric viruses in Burkinabe children.

  19. Identification of multiple novel viruses, including a parvovirus and a hepevirus, in feces of red foxes.

    Science.gov (United States)

    Bodewes, Rogier; van der Giessen, Joke; Haagmans, Bart L; Osterhaus, Albert D M E; Smits, Saskia L

    2013-07-01

    Red foxes (Vulpes vulpes) are the most widespread members of the order of Carnivora. Since they often live in (peri)urban areas, they are a potential reservoir of viruses that transmit from wildlife to humans or domestic animals. Here we evaluated the fecal viral microbiome of 13 red foxes by random PCR in combination with next-generation sequencing. Various novel viruses, including a parvovirus, bocavirus, adeno-associated virus, hepevirus, astroviruses, and picobirnaviruses, were identified.

  20. Viruses of foodborne origin: a review

    OpenAIRE

    Todd EC; Greig JD

    2015-01-01

    Ewen CD Todd,1,2 Judy D Greig3 1Ewen Todd Consulting LLC, Okemos, MI, USA; 2Department of Nutrition and Food Science, Faculty of Agricultural and Food Sciences, American University of Beirut, Beirut, Lebanon; 3Division of Public Health Risk Sciences, Laboratory for Foodborne Zoonoses, Public Health Agency of Canada, Guelph, ON, Canada Abstract: Enteric viruses are major contributors to foodborne disease, and include adenovirus, astrovirus, rotavirus, sapovirus, hepatitis A and E viruses, and...

  1. Epidemiology of gastroenteritis viruses in Japan: Prevalence, seasonality, and outbreak.

    Science.gov (United States)

    Thongprachum, Aksara; Khamrin, Pattara; Maneekarn, Niwat; Hayakawa, Satoshi; Ushijima, Hiroshi

    2016-04-01

    Acute gastroenteritis has been recognized as one of the most common diseases in humans and continues to be a major public health problem worldwide. Several groups of viruses have been reported as the causative agents of acute gastroenteritis, including rotavirus, norovirus, sapovirus, human astrovirus, adenovirus, and an increasing number of others which have been reported more recently. The epidemiology, prevalence, seasonality, and outbreaks of these viruses have been reviewed in a number of studies conducted in Japan over three decades. Rotavirus and norovirus were the two most common viruses detected almost equally in children under 5 years of age who were suffering from acute gastroenteritis. Like many other countries, the main rotavirus strains circulating in pediatric patients in Japan are G1P[8], G2P[4], G3P[8], and G9P[8]. Norovirus GII.4 was involved in most outbreaks in Japan and found to be associated with the emergence of new variants Sydney_2012. The classic human astrovirus, MLB, and VA clades astroviruses were also commonly found in pediatric patients with acute diarrhea. The sapovirus and adenovirus have been identified as the minor viral causative agents for acute gastroenteritis in Japan.

  2. A single-tube multiplex PCR for rapid detection in feces of 10 viruses causing diarrhea.

    Science.gov (United States)

    Khamrin, Pattara; Okame, Makiko; Thongprachum, Aksara; Nantachit, Nattika; Nishimura, Shuichi; Okitsu, Shoko; Maneekarn, Niwat; Ushijima, Hiroshi

    2011-05-01

    A novel multiplex polymerase chain reaction assay was developed to identify 10 viruses in a single tube. The assay was targeted to detect group A and C rotaviruses, adenovirus, norovirus GI, norovirus GII, sapovirus, astrovirus, Aichi virus, parechovirus, and enterovirus. A total of 235 stool samples were collected from infants and children with acute gastroenteritis in Kyoto, Japan, from 2008 to 2009, then tested by this novel multiplex PCR and compared with a multiplex PCR described previously, which used 3 primer sets. The novel multiplex PCR could detect the targeted viruses in 111 of the 235 (47.2%) stool samples. Of these, 9 out of 10 types of viruses were identified, including group A rotavirus, norovirus GII, enterovirus, sapovirus, adenovirus, parechovirus, group C rotavirus, astrovirus, and norovirus GI. In contrast, the multiplex PCR that used 3 sets of primers could detect the targeted viruses in 109 of the 235 (46.4%) stool samples. Among these, 8 types of viruses were identified, including group A rotavirus, norovirus GII, enterovirus, adenovirus, parechovirus, group C rotavirus, sapovirus, and astrovirus. The results suggested that the new multiplex PCR is useful as a rapid and cost effective diagnostic tool for the detection of major pathogenic viruses causing diarrhea.

  3. Enhanced enteric virus detection in sporadic gastroenteritis using a multi-target real-time PCR panel: a one-year study.

    Science.gov (United States)

    Pang, Xiaoli L; Preiksaitis, Jutta K; Lee, Bonita E

    2014-09-01

    Viral gastroenteritis causes significant mortality and morbidity worldwide. Identifying the etiology of viral gastroenteritis is a challenge as most enteric viruses (EVs) are non-culturable. This study is to develop an EV testing panel using real-time PCR (EVPrtPCR) to simultaneously detect rotavirus, norovirus, sapovirus, astrovirus, and enteric adenovirus in stool samples. EVPrtPCR using universal amplification conditions was run in a single instrument run. EVPrtPCR was used to test 2,486 sporadic gastroenteritis samples submitted for EV testing using electron microscopy (EM) between July 2008 and July 2009. Retesting spiked negative stool samples and Salmon DNA as internal control were used to evaluate inhibition. EVPrtPCR detected viruses in significantly more samples: 748 (34%) as compared to 94 (3.8%) by EM. EM did not detect any norovirus, sapovirus, and mixed infection, and detected only 39% of rotavirus and 38.2% of enteric adenovirus positive samples. Four samples that tested positive for rotavirus and two for adenovirus and for small-round-structured viruses by EM were negative by EVPrtPCR. Norovirus was the most common virus detected (17.6%) with 92.4% as genogroup II, followed by rotavirus (6.8%), sapovirus (4.2%), astrovirus (2.0%), and enteric adenovirus (1.4%) with 9% samples positive for mixed infection. Overall, EV identification followed a U-shaped age distribution; positive samples were more common in children ≤5 years old and adults >60 years old. Norovirus, sapovirus and astrovirus showed winter predominance and rotavirus peaked in the spring. No inhibition was observed. Molecular technology significantly enhanced the identification of EV causing sporadic gastroenteritis in Alberta.

  4. Feeding of the probiotic bacterium Enterococcus faecium NCIMB 10415 differentially affects shedding of enteric viruses in pigs

    Directory of Open Access Journals (Sweden)

    Kreuzer Susanne

    2012-07-01

    Full Text Available Abstract Effects of probiotic bacteria on viral infections have been described previously. Here, two groups of sows and their piglets were fed with or without feed supplementation of the probiotic bacterium Enterococcus faecium NCIMB 10415. Shedding of enteric viruses naturally occurring in these pigs was analyzed by quantitative real-time RT-PCR. No differences between the groups were recorded for hepatitis E virus, encephalomyocarditis virus and norovirus. In contrast, astrovirus was exclusively detected in the non-supplemented control group. Rotavirus was shedded later and with lower amounts in the probiotic piglet group (p p p p p p 

  5. LAS GASTROENTERITIS VÍRICAS: UN PROBLEMA DE SALUD PÚBLICA

    OpenAIRE

    Angela Domínguez; Pere Godoy; Núria Torner; Neus Cardeñosa; Ana Martínez

    2009-01-01

    Las gastroenteritis agudas (GA) constituyen un problema de salud importante, pudiendo estar causadas por bacterias, parásitos y virus enteropatógenos, entre los que se han reconocido hasta la actualidad rotavirus, astrovirus, adenovirus y calicivirus. La frecuencia de rotavirus como causa de casos esporádicos de GA oscila entre el 17,3% y el 37,4%. Aunque se ha sugerido que la exposición común no tiene mucha importancia para la difusión de este virus, resulta difícil relacionar casos aparente...

  6. High prevalence of enteric viruses in untreated individual drinking water sources and surface water in Slovenia.

    Science.gov (United States)

    Steyer, Andrej; Torkar, Karmen Godič; Gutiérrez-Aguirre, Ion; Poljšak-Prijatelj, Mateja

    2011-09-01

    Waterborne infections have been shown to be important in outbreaks of gastroenteritis throughout the world. Although improved sanitary conditions are being progressively applied, fecal contaminations remain an emerging problem also in developed countries. The aim of our study was to investigate the prevalence of fecal contaminated water sources in Slovenia, including surface waters and groundwater sources throughout the country. In total, 152 water samples were investigated, of which 72 samples represents groundwater from individual wells, 17 samples from public collection supplies and 63 samples from surface stream waters. Two liters of untreated water samples were collected and concentrated by the adsorption/elution technique with positively charged filters followed by an additional ultracentrifugation step. Group A rotaviruses, noroviruses (genogroups I and II) and astroviruses were detected with real-time RT-PCR method in 69 (45.4%) out of 152 samples collected, of which 31/89 (34.8%) drinking water and 38/63 (60.3%) surface water samples were positive for at least one virus tested. In 30.3% of drinking water samples group A rotaviruses were detected (27/89), followed by noroviruses GI (2.2%; 2/89) and astroviruses (2.2%; 2/89). In drinking groundwater samples group A rotaviruses were detected in 27 out of 72 tested samples (37.5%), genogroup I noroviruses in two (2.8%), and human astroviruses in one (1.4%) samples. In surface water samples norovirus genogroup GII was the most frequently detected (41.3%; 26/63), followed by norovirus GI (33.3%; 21/63), human astrovirus (27.0%; 17/63) and group A rotavirus (17.5%; 11/63). Our study demonstrates relatively high percentage of groundwater contamination in Slovenia and, suggests that raw groundwater used as individual drinking water supply may constitute a possible source of enteric virus infections. In the future, testing for enteric viruses should be applied for drinking water sources in waterborne outbreaks.

  7. [Viral diarrhea].

    Science.gov (United States)

    Stalder, H

    1982-04-12

    Viruses are one of the most frequent causes of acute infectious gastroenteritis. Thus, rotaviruses are responsible for 40-60%--and in winter sometimes for over 90% - of diarrhoea in infants. These viruses may also cause disease in adults, but small viruses of a size of about 27 nm, such as the Norwalk agent, play a much more important role. Several other non-cultivable viruses such as adeno-, calici- and astroviruses have been implicated as a cause of infectious diarrhoea. Knowledge of rotaviruses is so far advanced that routine laboratory diagnosis can easily be performed and active immunization against rotavirus infection envisaged.

  8. Genome Sequence Analysis of CsRV1: A Pathogenic Reovirus that Infects the Blue Crab Callinectes sapidus Across Its Trans-Hemispheric Range.

    Science.gov (United States)

    Flowers, Emily M; Bachvaroff, Tsvetan R; Warg, Janet V; Neill, John D; Killian, Mary L; Vinagre, Anapaula S; Brown, Shanai; Almeida, Andréa Santos E; Schott, Eric J

    2016-01-01

    The blue crab, Callinectes sapidus Rathbun, 1896, which is a commercially important trophic link in coastal ecosystems of the western Atlantic, is infected in both North and South America by C. sapidus Reovirus 1 (CsRV1), a double stranded RNA virus. The 12 genome segments of a North American strain of CsRV1 were sequenced using Ion Torrent technology. Putative functions could be assigned for 3 of the 13 proteins encoded in the genome, based on their similarity to proteins encoded in other reovirus genomes. Comparison of the CsRV1 RNA-dependent RNA polymerase (RdRP) sequence to genomes of other crab-infecting reoviruses shows that it is similar to the mud crab reovirus found in Scylla serrata and WX-2012 in Eriocheir sinensis, Chinese mitten crab, and supports the idea that there is a distinct "Crabreo" genus, different from Seadornavirus and Cardoreovirus, the two closest genera in the Reoviridae. A region of 98% nucleotide sequence identity between CsRV1 and the only available sequence of the P virus of Macropipus depurator suggests that these two viruses may be closely related. An 860 nucleotide region of the CsRV1 RdRP gene was amplified and sequenced from 15 infected crabs collected from across the geographic range of C. sapidus. Pairwise analysis of predicted protein sequences shows that CsRV1 strains in Brazil can be distinguished from those in North America based on conserved residues in this gene. The sequencing, annotation, and preliminary population metrics of the genome of CsRV1 should facilitate additional studies in diverse disciplines, including structure-function relationships of reovirus proteins, investigations into the evolution of the Reoviridae, and biogeographic research on the connectivity of C. sapidus populations across the Northern and Southern hemispheres.

  9. Comparative study of enteric viruses, coliphages and indicator bacteria for evaluating water quality in a tropical high-altitude system

    Directory of Open Access Journals (Sweden)

    Mazari-Hiriart Marisa

    2009-10-01

    Full Text Available Abstract Background Bacteria used as indicators for pathogenic microorganisms in water are not considered adequate as enteric virus indicators. Surface water from a tropical high-altitude system located in Mexico City that receives rainwater, treated and non-treated wastewater used for irrigation, and groundwater used for drinking, was studied. Methods The presence of enterovirus, rotavirus, astrovirus, coliphage, coliform bacteria, and enterococci was determined during annual cycles in 2001 and 2002. Enteric viruses in concentrated water samples were detected by reverse transcriptase-polymerase chain reaction (RT-PCR. Coliphages were detected using the double agar layer method. Bacteria analyses of the water samples were carried out by membrane filtration. Results The presence of viruses and bacteria in the water used for irrigation showed no relationship between current bacterial indicator detection and viral presence. Coliphages showed strong association with indicator bacteria and enterovirus, but weak association with other enteric viruses. Enterovirus and rotavirus showed significant seasonal differences in water used for irrigation, although this was not clear for astrovirus. Conclusion Coliphages proved to be adequate faecal pollution indicators for the irrigation water studied. Viral presence in this tropical high-altitude system showed a similar trend to data previously reported for temperate zones.

  10. Incidence of Norovirus and Other Viral Pathogens That Cause Acute Gastroenteritis (AGE) among Kaiser Permanente Member Populations in the United States, 2012-2013.

    Science.gov (United States)

    Grytdal, Scott P; DeBess, Emilio; Lee, Lore E; Blythe, David; Ryan, Patricia; Biggs, Christianne; Cameron, Miriam; Schmidt, Mark; Parashar, Umesh D; Hall, Aron J

    2016-01-01

    Noroviruses and other viral pathogens are increasingly recognized as frequent causes of acute gastroenteritis (AGE). However, few laboratory-based data are available on the incidence of AGE caused by viral pathogens in the U.S. This study examined stool specimens submitted for routine clinical diagnostics from patients enrolled in Kaiser Permanente (KP) health plans in metro Portland, OR, and the Maryland, District of Columbia, and northern Virginia geographic areas to estimate the incidence of viral enteropathogens in these populations. Over a one-year study period, participating laboratories randomly selected stools submitted for routine clinical diagnostics for inclusion in the study along with accompanying demographic and clinical data. Selected stools were tested for norovirus, rotavirus, sapovirus, and astrovirus using standardized real-time RT-PCR protocols. Each KP site provided administrative data which were used in conjunction with previously published data on healthcare utilization to extrapolate pathogen detection rates into population-based incidence rates. A total of 1,099 specimens collected during August 2012 to September 2013 were included. Mean age of patients providing stool specimens was 46 years (range: 0-98 years). Noroviruses were the most common viral pathogen identified among patients with AGE (n = 63 specimens, 6% of specimens tested). In addition, 22 (2%) of specimens were positive for rotavirus; 19 (2%) were positive for sapovirus; and 7 (1%) were positive for astrovirus. Incidence of norovirus-associated outpatient visits was 5.6 per 1,000 person-years; incidence of norovirus disease in the community was estimated to be 69.5 per 1,000 person-years. Norovirus incidence was highest among children 65 years (outpatient incidence = 7.8 per 1,000 person-years; community incidence = 75.8 per 1,000 person-years). Outpatient incidence rates of rotavirus, sapovirus, and astrovirus were 2.0, 1.6, 0.6 per 1,000 person-years, respectively; community

  11. Incidence of Norovirus and Other Viral Pathogens That Cause Acute Gastroenteritis (AGE among Kaiser Permanente Member Populations in the United States, 2012-2013.

    Directory of Open Access Journals (Sweden)

    Scott P Grytdal

    Full Text Available Noroviruses and other viral pathogens are increasingly recognized as frequent causes of acute gastroenteritis (AGE. However, few laboratory-based data are available on the incidence of AGE caused by viral pathogens in the U.S. This study examined stool specimens submitted for routine clinical diagnostics from patients enrolled in Kaiser Permanente (KP health plans in metro Portland, OR, and the Maryland, District of Columbia, and northern Virginia geographic areas to estimate the incidence of viral enteropathogens in these populations. Over a one-year study period, participating laboratories randomly selected stools submitted for routine clinical diagnostics for inclusion in the study along with accompanying demographic and clinical data. Selected stools were tested for norovirus, rotavirus, sapovirus, and astrovirus using standardized real-time RT-PCR protocols. Each KP site provided administrative data which were used in conjunction with previously published data on healthcare utilization to extrapolate pathogen detection rates into population-based incidence rates. A total of 1,099 specimens collected during August 2012 to September 2013 were included. Mean age of patients providing stool specimens was 46 years (range: 0-98 years. Noroviruses were the most common viral pathogen identified among patients with AGE (n = 63 specimens, 6% of specimens tested. In addition, 22 (2% of specimens were positive for rotavirus; 19 (2% were positive for sapovirus; and 7 (1% were positive for astrovirus. Incidence of norovirus-associated outpatient visits was 5.6 per 1,000 person-years; incidence of norovirus disease in the community was estimated to be 69.5 per 1,000 person-years. Norovirus incidence was highest among children 65 years (outpatient incidence = 7.8 per 1,000 person-years; community incidence = 75.8 per 1,000 person-years. Outpatient incidence rates of rotavirus, sapovirus, and astrovirus were 2.0, 1.6, 0.6 per 1,000 person

  12. Use of dual priming oligonucleotide system-based multiplex RT-PCR combined with high performance liquid chromatography assay for simultaneous detection of five enteric viruses associated with acute enteritis.

    Science.gov (United States)

    Fan, Wen-Lu; Wang, Zi-Wei; Qin, Yue; Sun, Chao; Liu, Zhong-Mei; Jiang, Yan-Ping; Qiao, Xin-Yuan; Tang, Li-Jie; Li, Yi-Jing; Xu, Yi-Gang

    2017-05-01

    In this study, a specific and sensitive method for simultaneous detection of human astrovirus, human rotavirus, norovirus, sapovirus and enteric adenovirus associated with acute enteritis was developed, based on the specific dual priming oligonucleotide (DPO) system and the sensitive high-performance liquid chromatography (HPLC) analysis. The DPO system-based multiplex reverse transcription-polymerase chain reaction (RT-PCR) combined with HPLC assay was more sensitive than agarose gel electrophoresis analysis and real-time SYBR Green PCR assay, and showed a specificity of 100% and sensitivity of 96%-100%. The high sensitivity and specificity of the assay indicates its great potential to be a useful tool for the accurate diagnosis of enteric virus infections.

  13. Acute diarrhea in West African children: diverse enteric viruses and a novel parvovirus genus.

    Science.gov (United States)

    Phan, Tung G; Vo, Nguyen P; Bonkoungou, Isidore J O; Kapoor, Amit; Barro, Nicolas; O'Ryan, Miguel; Kapusinszky, Beatrix; Wang, Chunling; Delwart, Eric

    2012-10-01

    Parvoviruses cause a variety of mild to severe symptoms or asymptomatic infections in humans and animals. During a viral metagenomic analysis of feces from children with acute diarrhea in Burkina Faso, we identified in decreasing prevalence nucleic acids from anelloviruses, dependoviruses, sapoviruses, enteroviruses, bocaviruses, noroviruses, adenoviruses, parechoviruses, rotaviruses, cosavirus, astroviruses, and hepatitis B virus. Sequences from a highly divergent parvovirus, provisionally called bufavirus, were also detected whose NS1 and VP1 proteins showed parvoviruses. Four percent of the fecal samples were PCR positive for this new parvovirus, including a related bufavirus species showing only 72% identity in VP1. The high degree of genetic divergence of these related genomes from those of other parvoviruses indicates the presence of a proposed new Parvoviridae genus containing at least two species. Studies of the tropism and pathogenicity of these novel parvoviruses will be facilitated by the availability of their genome sequences.

  14. Identification of GBV-D, a novel GB-like flavivirus from old world frugivorous bats (Pteropus giganteus in Bangladesh.

    Directory of Open Access Journals (Sweden)

    Jonathan H Epstein

    Full Text Available Bats are reservoirs for a wide range of zoonotic agents including lyssa-, henipah-, SARS-like corona-, Marburg-, Ebola-, and astroviruses. In an effort to survey for the presence of other infectious agents, known and unknown, we screened sera from 16 Pteropus giganteus bats from Faridpur, Bangladesh, using high-throughput pyrosequencing. Sequence analyses indicated the presence of a previously undescribed virus that has approximately 50% identity at the amino acid level to GB virus A and C (GBV-A and -C. Viral nucleic acid was present in 5 of 98 sera (5% from a single colony of free-ranging bats. Infection was not associated with evidence of hepatitis or hepatic dysfunction. Phylogenetic analysis indicates that this first GBV-like flavivirus reported in bats constitutes a distinct species within the Flaviviridae family and is ancestral to the GBV-A and -C virus clades.

  15. Etiology of diarrhea in young children in Denmark: a case-control study

    DEFF Research Database (Denmark)

    Olesen, B.; Neimann, J.; Bottiger, B.

    2005-01-01

    -matched controls were examined, and their parents were interviewed concerning symptoms. Rotavirus, adenovirus, and astrovirus were detected by enzyme-linked immunosorbent assay, and norovirus and sapovirus were detected by PCR. Salmonella, thermotolerant Campylobacter, Yersinia, Shigella, and Vibrio spp. were...... by microscopy. Overall, a potential pathogen was found in 54% of cases. More cases than controls were infected with rotavirus, Salmonella, norovirus, adenovirus, Campylobacter, sapovirus, STEC, classical EPEC, Yersinia, and Cryptosporidium strains, whereas A/EEC, although common, was not associated with illness....... The single most important cause of diarrhea was rotavirus, which points toward the need for a childhood vaccine for this pathogen, but norovirus, adenovirus, and sapovirus were also major etiologies. Salmonella sp. was the most common bacterial pathogen, followed by Campylobacter, STEC, Yersinia...

  16. Viral immunity. Transkingdom control of viral infection and immunity in the mammalian intestine.

    Science.gov (United States)

    Pfeiffer, Julie K; Virgin, Herbert W

    2016-01-15

    Viruses that infect the intestine include major human pathogens (retroviruses, noroviruses, rotaviruses, astroviruses, picornaviruses, adenoviruses, herpesviruses) that constitute a serious public health problem worldwide. These viral pathogens are members of a large, complex viral community inhabiting the intestine termed "the enteric virome." Enteric viruses have intimate functional and genetic relationships with both the host and other microbial constituents that inhabit the intestine, such as the bacterial microbiota, their associated phages, helminthes, and fungi, which together constitute the microbiome. Emerging data indicate that enteric viruses regulate, and are in turn regulated by, these other microbes through a series of processes termed "transkingdom interactions." This represents a changing paradigm in intestinal immunity to viral infection. Here we review recent advances in the field and propose new ways in which to conceptualize this important area.

  17. Acute viral gastroenteritis in children hospitalized in Iksan, Korea during December 2010 - June 2011

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    Cheol Whoan So

    2013-09-01

    Full Text Available Purpose: Viral etiology is common in cases of children with acute diarrhea, and antibiotic therapy is usually not required. Therefore, it is important to determine the distribution of common viruses among children hospitalized with acute diarrhea. Methods: We included 186 children who suffered from acute diarrhea and were hospitalized at the Wonkwang University Hospital Pediatric ward from December 1, 2010 to June 30, 2011 in this study. Stool samples were collected and multiplex reverse transcriptase polymerase chain reaction (multiplex RT-PCR was used to simultaneously determine the viral etiology such as rotavirus, norovirus, astrovirus, or adenovirus.&lt;br&gt; Results: Causative viruses were detected in 72 of the 186 cases (38.7%. The mean age of the viruspositive cases was 1 year and 9 months (range, 1 month to 11 years. Rotavirus was detected in 50/186 (26.9%; norovirus, in 18/186 (9.7%; and astrovirus, in 3/186 cases (1.6%. Adenovirus was not detected in any of the cases. Proportions of norovirus genogroups I and II were 21.1% and 78.9%, respectively. Four of the 51 rotavirus-positive cases (7.8% had received rotavirus vaccination at least once. The mean duration of diarrhea was 2.8 days (range, 1 to 10 days and vomiting occurred in 39 of the 72 cases (54.2%.&lt;br&gt; Conclusion: Viral etiology was confirmed in about one-third of the children with acute diarrhea, and the most common viral agent was rotavirus, followed by norovirus.

  18. Evaluation of the results of acute viral gastroenteritis data in Refik Saydam National Public Health Agency, Virology Reference and Research Laboratory in 2009

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    Nurhan ALBAYRAK

    2011-01-01

    Full Text Available Objective: Norovirus, Rotavirus, Adenovirus and Astrovirus are responsible for most non-bacterial acute gastroenteritis. The incidence of these viral agents in Turkey is not well known. In this study, it was aimed to document the viral etiology of the stool samples which were send to Refik Saydam National Public Health Agency (RSNPHA, Virology Reference and Research Laboratory for investigation of viral acute gastroenteritis agents. Method: A total of 147 stool samples from 11 different provinces were send to the Virology Laboratory for Reference and Research of RSNPHA in 2009. Samples were collected from patients admitted because of acute gastroenteritis and from the cases with the signs of illness at different times of the year and sent by the Provincial Health Directorates to our laboratory. The samples were examined in the laboratory using the commercial multiplex real-time PCR kit for norovirus genotype I, norovirus genotype II, rotavirus, adenovirus and astrovirus. Results: 65 (44.2 % samples were found to be positive at least for one viral agent and 10 (6.8 % samples for more than one viral agent. Norovirus (particularly genotype II infections were detected as the most prevalent viral agent in acute gastroenteritis patients in this period. Rotavirus infections were determined as the second most common infection after norovirus infections. Adenovirus infections have been found to be the least prevalent agent in the laboratory. Conclusion: Results of this study showed that norovirus genotype II has been more commonly responsible for acute diarrhea than the other viral pathogens. The viral agents we have studied should be considered as pathogens that can be seen in all seasons. Viral factors should not be underestimated as the cause of acute gastroenteritis; additionally it should be noted that acute gastroenteritis could be caused by coinfection of viral agents.

  19. Simultaneous investigation of influenza and enteric viruses in the stools of adult patients consulting in general practice for acute diarrhea

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    Arena Christophe

    2012-06-01

    Full Text Available Abstract Background Gastrointestinal symptoms are not an uncommon manifestation of an influenza virus infection. In the present study, we aimed to investigate the presence of influenza viruses in the stools of adult patients consulting their general practitioner for uncomplicated acute diarrhea (AD and the proportion of concurrent infections by enteric and influenza viruses. Method A case-control study was conducted from December 2010 to April 2011. Stool specimens were collected and tested for influenza viruses A (seasonal A/H3N2 and pandemic A/H1N1 and B, and for four enteric viruses (astrovirus, group A rotavirus, human enteric adenovirus, norovirus of genogroups I – NoVGI - and genogroup II - NoVGII. Results General practitioners enrolled 138 cases and 93 controls. Of the 138 stool specimens collected, 92 (66.7% were positive for at least one of the four enteric viruses analysed and 10 (7.2% tested positive for one influenza virus. None of these 10 influenza positive patients reported respiratory symptoms. In five influenza-positive patients (3.6%, we also detected one enteric virus, with 4 of them being positive for influenza B (2 had co-detection with NoVGI, 1 with NoVGII, and 1 with astrovirus. None of the 93 controls tested positive for one of the enteric and/or other influenza viruses we investigated. Conclusions In this study we showed that the simultaneous detection of influenza and enteric viruses is not a rare event. We have also reported, for the first time in general practice, the presence of seasonal and pandemic influenza viruses in the stools of adult patients consulting for uncomplicated AD. A simultaneous investigation of enteric and influenza viruses in patients complaining of gastrointestinal symptoms could be useful for future studies to better identify the agents responsible for AD.

  20. Development and application of an RT-PCR test for detecting avian nephritis virus.

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    Todd, D; Trudgett, J; McNeilly, F; McBride, N; Donnelly, B; Smyth, V J; Jewhurst, H L; Adair, B M

    2010-06-01

    The development of a reverse transcriptase-polymerase chain reaction (RT-PCR) test for detecting avian nephritis virus (ANV) is described. Primers, which amplified a fragment of 182 base pairs (bp), were located in the conserved 3' untranslated region (UTR) of the genome. The limit of detection of the test was estimated to be approximately 18 viral copies using a 10-fold dilution series of in vitro transcribed RNA. Positive signals were produced with representative ANV samples, some of which were not detected by previously described RT-PCR tests for detecting ANV, but other avian astroviruses including chicken astrovirus isolates and duck hepatitis virus types 2 and 3 tested negative. When applied to gut content samples from UK, German and US broiler flocks with enteritis/growth problems, ANVs were detected by RT-PCR in 82/82 (100%) samples. ANVs were also detected in 80/96 (83%) pooled gut content samples from longitudinal surveys of four broiler flocks displaying below-average performance. Whereas all samples collected on day 0 from the surveys were negative for ANV, all samples collected at days 4/5, 7, 10, 14, 21 and 28 tested positive. Sequence determinations performed with amplicons produced with 14 field samples confirmed the ANV specificity of the test, while comparative and phylogenetic analyses based on 109-nucleotide 3'-UTR sequences demonstrated that the majority of ANVs investigated were more closely related to the serotype 2 ANV (accession number AB 046864) than to the serotype 1 ANV (accession number NC 003790).

  1. LAS GASTROENTERITIS VÍRICAS: UN PROBLEMA DE SALUD PÚBLICA

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    Angela Domínguez

    2009-01-01

    Full Text Available Las gastroenteritis agudas (GA constituyen un problema de salud importante, pudiendo estar causadas por bacterias, parásitos y virus enteropatógenos, entre los que se han reconocido hasta la actualidad rotavirus, astrovirus, adenovirus y calicivirus. La frecuencia de rotavirus como causa de casos esporádicos de GA oscila entre el 17,3% y el 37,4%. Aunque se ha sugerido que la exposición común no tiene mucha importancia para la difusión de este virus, resulta difícil relacionar casos aparentemente esporádicos, por lo que probablemente haya brotes que no llegan a detectarse. Los astrovirus causan GA con una frecuencia que oscila entre un 2 y un 26% y se han descrito brotes en escuelas y guarderías, pero también en población adulta y en ancianos. La frecuencia con que se identifican los adenovirus 40 y 41 como causa de GA esporádica en niños no inmunodeprimidos oscila entre 0,7% y 31,5%, aunque probablemente haya subdetección debido a que la sensibilidad de las técnicas convencionales es baja. Los calicivirus se separan filogenéticamente en dos géneros: Norovirus y Sapovirus. Los Norovirus se asocian frecuentemente con brotes de GA transmitidos por agua y alimentos . Se estima que el 40% de los casos de GA por Norovirus están vehiculados por alimentos. En Suecia y en algunas zonas de Estados Unidos es la primera causa de brotes de toxiinfecciones alimentarias. Recientemente se han descrito en Canadà y Japón brotes de transmisión persona a persona y también de transmisión alimentaria causados por Sapovirus, afectando tanto a población infantil como adulta.

  2. ACUTE ENTERIC INFECTIONS POLYMERASE CHAIN REACTION ASSAY IN PEDIATRIC PRACTICE: OPPORTUNITIES AND CHALLENGES

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    E. D. Sokolova

    2016-01-01

    Full Text Available The aim of the study is estimate the opportunities of local multi-prime PCR reagents kits in children enteric infections etiological diagnostics amongst the patients with diarrhoea vs traditional bacteriological methods. We used 4 kits of reagents that provide multiple pathogens simultaneous indication in one sample: 1 Rotavirus, Norovirus, Astrovirus; 2 Shigella spp./EIEC, Salmonella spp., Campylobacter spp.; 3 Yersinia enterocolitica and Yersinia pseudotuberculosis; 4 E. coli: EIEC (enteroinvasive, EPEC (enteropathogenic, ETEC (enterotoxigenic, EHEC (enterohaemorrhagic, EAgEC (enteroaggregative. It has been shown that the viral intestinal infections is increased by 14%, bacterial — in 2,5 times. PCR diagnostics identified in 62% of patients the viral gastroenteritis: Rotavirus (52%, Norovirus (9%, Astrovirus (1%. Detected bacterial pathogens PCR markers number proved up to 2.5 times high than according to bacteriological examination. The spectrum of bacterial agents increased due to E. coli and Y. enterocolitica. PCR diagnostics increased detection of Campylobacter up to 2 times. Detected E. coli DNA prevalence: EPEC — 66%, EAgEC, ETEC and EHEC were 31%, 9% and 4%, respectively. DNA Campylobacter spp. and E. coli constituted 2/3 of all findings: Campylobacter spp. (41%, E. coli (24%, Salmonella spp. (19%, Yersinia spp. (11%, Shigella spp./EIEC (5%. The positive results of bacteriological and serological methods duplicate the positive results of PCR diagnostics. In general, the positive results of PCR diagnosis of bacterial pathogens were detected in 46.35% of the examined patients. In 48.4% of patients identified PCR markers viral — bacterial infection, in 5.25% — of bacterial associations, in 11% of them were found the DNA 2–3 bacterial pathogens. The study was shown in children in St. Petersburg in 2012–2014 dominated rotavirus infection, campylobacteriosis and escherichiosis. The prevalence of viral-bacterial confections is more

  3. Microbial impact of small tributaries on water and shellfish quality in shallow coastal areas.

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    Riou, P; Le Saux, J C; Dumas, F; Caprais, M P; Le Guyader, S F; Pommepuy, M

    2007-06-01

    This study aimed to investigate the impact of small tributaries on seawater and shellfish quality in coastal area subjected to brief episodes leading to fecal contamination. Escherichia coli and F-RNA-specific bacteriophages were selected as fecal indicators and astroviruses were chosen as being representative of pathogens in the human population during winter viral epidemics. A two-dimensional hydrodynamic model was built to simulate the current and dispersion in the model domain, which includes areas uncovered at low tide. The model also includes decay rates to simulate microorganism behavior and assess the influence of fecal input on shellfish quality. The originality lies in the fact that specific features of the study area were considered. Modeling results indicate limited particle movements and long flushing times at the back of the bay, where shellfish are farmed. Computational results showed that under normal conditions, i.e. 94% of the time, when rainfall was less than 10 mm per day, the sector shows acceptable water quality. These results are in agreement with shellfish concentration measured in the field. Under high flow conditions, high concentrations of fecal indicators and astrovirus were measured in the river and tributaries. The corresponding fluxes were over 50 times higher than under normal weather conditions. The location of the shellfish beds near the coast makes them vulnerable and fecal indicators and viruses were detected in shellfish after short rainfall events. Our modeling approach makes a contribution to shellfish management and consumer protection, by indicating the "risk period" as defined by EU regulations. Molecular development such as viral quantification in conjunction with model developments will help to prevent shellfish contamination and thus provide safer products to consumers and an effective tool for shellfish producers.

  4. Screening of the Host Target Proteins of HAstV nsP1 a/1%人星状病毒非结构蛋白nsP1 a/1相互作用的宿主细胞靶蛋白筛选

    Institute of Scientific and Technical Information of China (English)

    刘志成; 苑荣亮; 张成; 汪玉婷; 黄芬; 井申荣

    2016-01-01

    Objective To screen out the host target proteins that can interact with the human astrovirus nonstructural protein nsP1 a/1 ( HAstV nsP1 a/1 ) by utilizing the bacterial two⁃hybrid system. Methods The genome was extracted from cultured HT29 cells. HT29 genomic library was constructed by using pUT18 C, which was named HT29⁃pUT18 C. Then the carrier of bait nsP1a/1⁃pKT25 was established as well. Both the genomic library and the carrier of bait were transformed into BTH101 , which was followed by screening on M63 culture medium. Results The structure of the bait carrier was demonstrated to be right by the enzyme digestion method and the sequencing analy⁃sis. The host target proteins that had interaction with HAstV nsP1 a/1 were successfully isolated by u⁃tilizing the bacerial two⁃hybrid system. Conclusion The bait carrier of HAstV nsP1 a/1 was suc⁃cessfully established, and three proteins that could interact with nsP1a/1 were identified in the HT29 genomic library, which lay a foundation for further researches on the cellular proliferation of human astroviruses and application of drugs for the prevention and treatment of human astroviruse in⁃fections.%目的:利用细菌双杂交技术筛选与人星状病毒非结构蛋白nsP1a/1相互作用的蛋白。方法培养人肠道细胞HT29,提取基因组,利用细菌双杂交系统载体pUT18 C构建HT29基因组文库,同时构建诱饵载体nsP1a/1⁃pKT25。将文库和诱饵载体共转化进入细菌BTH101,在M63培养基上进行筛选。结果通过酶切鉴定以及测序分析,诱饵载体构建正确,并且利用细菌双杂交技术成功从HT29细胞基因组文库中筛选出能与人星状病毒非结构蛋白 nsP1a/1相互作用的蛋白。结论成功构建了人星状病毒非结构蛋白nsP1a/1的诱饵载体,在HT29细胞基因组文库中筛选得到了3个与nsP1a/1相互作用的蛋白,为进一步研究人星状病毒在细胞内的增殖和防治药物的应用奠定基础。

  5. Hepatitis Virus Infections in Poultry.

    Science.gov (United States)

    Yugo, Danielle M; Hauck, Ruediger; Shivaprasad, H L; Meng, Xiang-Jin

    2016-09-01

    Viral hepatitis in poultry is a complex disease syndrome caused by several viruses belonging to different families including avian hepatitis E virus (HEV), duck hepatitis B virus (DHBV), duck hepatitis A virus (DHAV-1, -2, -3), duck hepatitis virus Types 2 and 3, fowl adenoviruses (FAdV), and turkey hepatitis virus (THV). While these hepatitis viruses share the same target organ, the liver, they each possess unique clinical and biological features. In this article, we aim to review the common and unique features of major poultry hepatitis viruses in an effort to identify the knowledge gaps and aid the prevention and control of poultry viral hepatitis. Avian HEV is an Orthohepevirus B in the family Hepeviridae that naturally infects chickens and consists of three distinct genotypes worldwide. Avian HEV is associated with hepatitis-splenomegaly syndrome or big liver and spleen disease in chickens, although the majority of the infected birds are subclinical. Avihepadnaviruses in the family of Hepadnaviridae have been isolated from ducks, snow geese, white storks, grey herons, cranes, and parrots. DHBV evolved with the host as a noncytopathic form without clinical signs and rarely progressed to chronicity. The outcome for DHBV infection varies by the host's ability to elicit an immune response and is dose and age dependent in ducks, thus mimicking the pathogenesis of human hepatitis B virus (HBV) infections and providing an excellent animal model for human HBV. DHAV is a picornavirus that causes a highly contagious virus infection in ducks with up to 100% flock mortality in ducklings under 6 wk of age, while older birds remain unaffected. The high morbidity and mortality has an economic impact on intensive duck production farming. Duck hepatitis virus Types 2 and 3 are astroviruses in the family of Astroviridae with similarity phylogenetically to turkey astroviruses, implicating the potential for cross-species infections between strains. Duck astrovirus (DAstV) causes

  6. Identificación y caracterización genética de agentes virales diarreagénicos en niños menores de cinco años de edad con o sin diarrea en Bucaramanga

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    Nayibe Tatiana Sánchez Álvarez

    2016-06-01

    Full Text Available Introducción: La enfermedad diarreica aguda (EDA, es una de las principales causas de morbi-mortalidad en todos los grupos de edad, con mayor incidencia en niños menores de cinco años. EDA es causada por diversos agentes. Entre los que se asocian de tipo bacteriano, parasitario y principalmente viral. La epidemiología y el diagnóstico en las infecciones virales es limitado debido a los altos costos en las técnicas moleculares y al desconocimiento por parte del personal de salud sobre su importancia. En Colombia son escasos los estudios reportados sobre la prevalencia de las infecciones virales diarreagénicas. Objetivo: Identificar la frecuencia de agentes virales asociados a las enfermedades diarreicas y caracterizar el agente viral más en los niveles genéticos y filogenéticos. Materiales y métodos: Este estudio hace parte de un estudio de casos y controles aprobado por el National Institutes of Health (NIH, en esta investigación se incluyeron 405 niños con EDA y 405 controles de niños sanos. Las muestras de heces se procesaron por métodos moleculares basados en la detección de adenovirus, norovirus, sapovirus, y astrovirus. Rotavirus se detectó mediante un ensayo de ELISA convencional. Análisis filogenético se llevó a cabo tanto para rotavirus como norovirus, mediante la amplificación de secuencia de ADN de los fragmentos de la región C de la cápside y detección de las proteínas VP4 y VP7 respectivamente. Las secuencias para cada tipo de virus se compararon y se construyó un árbol filogenético utilizando secuencias de referencia. Resultados: En general 243 virus fueron detectados entre las 810 muestras, 183 en casos y 60 en controles. El virus con mayor prevalencia fue norovirus GII con 10.9 %, seguido de rotavirus sapovirus con 5.1 %. Norovirus GI, astrovirus y adenovirus fueron identificados en 3.8 %, 3.2 % y 1.7 % respectivamente. La positividad fue más frecuente en el rango de edad  de 12 a 23 meses y de 24 a

  7. Etiology and Epidemiology of Diarrhea in Hospitalized Children from Low Income Country: A Matched Case-Control Study in Central African Republic.

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    Sébastien Breurec

    2016-01-01

    Full Text Available In Sub-Saharan Africa, infectious diarrhea is a major cause of morbidity and mortality. A case-control study was conducted to identify the etiology of diarrhea and to describe its main epidemiologic risk factors among hospitalized children under five years old in Bangui, Central African Republic.All consecutive children under five years old hospitalized for diarrhea in the Pediatric Complex of Bangui for whom a parent's written consent was provided were included. Controls matched by age, sex and neighborhood of residence of each case were included. For both cases and controls, demographic, socio-economic and anthropometric data were recorded. Stool samples were collected to identify enteropathogens at enrollment. Clinical examination data and blood samples were collected only for cases.A total of 333 cases and 333 controls was recruited between December 2011 and November 2013. The mean age of cases was 12.9 months, and 56% were male. The mean delay between the onset of first symptoms and hospital admission was 3.7 days. Blood was detected in 5% of stool samples from cases. Cases were significantly more severely or moderately malnourished than controls. One of the sought-for pathogens was identified in 78% and 40% of cases and controls, respectively. Most attributable cases of hospitalized diarrhea were due to rotavirus, with an attributable fraction of 39%. Four other pathogens were associated with hospitalized diarrhea: Shigella/EIEC, Cryptosporidium parvum/hominis, astrovirus and norovirus with attributable fraction of 9%, 10%, 7% and 7% respectively. Giardia intestinalis was found in more controls than cases, with a protective fraction of 6%.Rotavirus, norovirus, astrovirus, Shigella/EIEC, Cryptosporidium parvum/hominis were found to be positively associated with severe diarrhea: while Giardia intestinalis was found negatively associated. Most attributable episodes of severe diarrhea were associated with rotavirus, highlighting the urgent

  8. Microchip capillary electrophoresis with laser-induced fluorescence combined with one-step duplex reverse-transcription polymerase chain reaction for the rapid detection of Enterovirus 71 and Coxsackievirus A16 in throat swab specimens.

    Science.gov (United States)

    Jia, Ruan; Chengjun, Sun; Heng, Chen; Chen, Zhou; Yuanqian, Li; Yongxin, Li

    2015-07-01

    Enterovirus 71 and Coxsackievirus A16 are the main pathogens causing hand-foot-mouth disease. In this paper, microchip capillary electrophoresis with laser-induced fluorescence combined with one-step duplex reverse transcript-polymerase chain reaction has been developed for the detection of Enterovirus 71 and Coxsackievirus A16 in throat swab specimens. The specific reverse transcription-polymerase chain reaction amplicons labeled with SYBR Orange were separated by microchip capillary electrophoresis and detected by laser induced fluorescence detector within 7 min. The intraday and interday relative standard deviation of migration time for DNA Marker was in the range of 1.36-2.94 and 2.78-3.96%, respectively. The detection limits were as low as 2.06 × 10(3) copies/mL for Enterovirus 71 and 5 × 10(3) copies/mL for Coxsackievirus A16. No cross-reactivity was observed with rotavirus, astrovirus, norovirus, and adenovirus, which showed good specificity of the method. This assay was validated using 100 throat swab specimens that were detected by real-time reverse-transcript polymerase chain reaction in parallel and the two methods produced the same results. This study provided a rapid, sensitive and specific method for the detection of Enterovirus 71 and Coxsackievirus A16, which make a contribution to significant time and cost saving for the identification and treatment of patients.

  9. Molecular surveillance of non-polio enterovirus infections in patients with acute gastroenteritis in Western India: 2004-2009.

    Science.gov (United States)

    Patil, Pooja R; Chitambar, Shobha D; Gopalkrishna, V

    2015-01-01

    Acute gastroenteritis is a major cause of childhood morbidity and mortality worldwide. Rotavirus (RV) and Norovirus (NoV) are the leading cause of the disease. Despite the use of improved diagnostic methods a significant proportion of gastroenteritis cases remained undiagnosed. Though nonpolio enteroviruses (NPEVs) have been reported frequently in children with acute gastroenteritis, their etiologic role has not been established. To investigate the epidemiology of NPEVs in gastroenteritis cases which remained negative for leading causative agents, 955 RV and NoV negative stool specimens from children hospitalized for acute gastroenteritis were included in the study. A case control study was conducted which includes stool specimens from 450 children with gastroenteritis and 162 asymptomatic control subjects to determine the association of NPEVs with the disease. NPEV detection and typing was carried out by RT-PCR and sequencing. Presence of RV, NoV, Adenovirus, and Astrovirus was confirmed by ELISA or PCR/RT-PCR. Overall 14% NPEV prevalence was noted. The percentage of children with NPEV infection differed significantly between gastroenteritis and non-gastroenteritis patients (13.7% vs. 4.9%). NPEV was more prevalent among patients with gastroenteritis of undetectable etiology as compared to those detected positive for other viruses (17.9% vs. 7%) (P gastroenteritis and highlights the clinical and epidemiological features of NPEV infections in patients with acute gastroenteritis from western India.

  10. Molecular detection of chicken parvovirus in broilers with enteric disorders presenting curving of duodenal loop, pancreatic atrophy, and mesenteritis.

    Science.gov (United States)

    Nuñez, L F N; Sá, L R M; Parra, S H S; Astolfi-Ferreira, C S; Carranza, C; Ferreira, A J P

    2016-04-01

    Enteric disorders are an important cause of economic losses in broiler chickens worldwide. Several agents have been associated with enteric problems, such as viruses, bacteria, and parasites. In this study, broiler chickens showing signs of enteric disorders were subjected to molecular diagnosis for several viral agents and also for pathological examination for elucidating this problem. Thus, the chickens were screened for avian nephritis virus (ANV), chicken astrovirus (CAstV), avian rotavirus (ArtV), avian reovirus (AReoV), infectious bronchitis virus (IBV), fowl adenovirus group I (FAdV-1), and chicken parvovirus (ChPV). Postmortem examinations revealed a curving of the duodenal loop (J-like appearance) and intestines filled with liquid and gaseous content. Histopathological analysis of the duodenal loop showed pancreatic atrophy, acute mesenteritis, and enteritis. PCR results showed that ChPV was the sole viral agent detected in samples with lesions such as the curved duodenal loop and pancreatic atrophy. Molecular characterization of the nucleotide and deduced amino acid sequences revealed a high similarity with other strains of ChPV from Brazil, Canada, United States, Europe, and Asia. These findings suggest an association between ChPV and the development of enteritis, pancreatitis, and pancreatic atrophy, which may lead to curling of the duodenal loop. Together, these alterations may disrupt the normal functioning of the digestive system, diminishing digestion and the absorption of dietary nutrients and consequently leading to reduced weight gain, flock impairment, dwarfism, and an elevated feed conversion rate.

  11. Viral infections of rabbits.

    Science.gov (United States)

    Kerr, Peter J; Donnelly, Thomas M

    2013-05-01

    Viral diseases of rabbits have been used historically to study oncogenesis (e.g. rabbit fibroma virus, cottontail rabbit papillomavirus) and biologically to control feral rabbit populations (e.g. myxoma virus). However, clinicians seeing pet rabbits in North America infrequently encounter viral diseases although myxomatosis may be seen occasionally. The situation is different in Europe and Australia, where myxomatosis and rabbit hemorrhagic disease are endemic. Advances in epidemiology and virology have led to detection of other lapine viruses that are now recognized as agents of emerging infectious diseases. Rabbit caliciviruses, related to rabbit hemorrhagic disease, are generally avirulent, but lethal variants are being identified in Europe and North America. Enteric viruses including lapine rotavirus, rabbit enteric coronavirus and rabbit astrovirus are being acknowledged as contributors to the multifactorial enteritis complex of juvenile rabbits. Three avirulent leporid herpesviruses are found in domestic rabbits. A fourth highly pathogenic virus designated leporid herpesvirus 4 has been described in Canada and Alaska. This review considers viruses affecting rabbits by their clinical significance. Viruses of major and minor clinical significance are described, and viruses of laboratory significance are mentioned.

  12. Surveillance of Food- and Smear-Transmitted Pathogens in European Soldiers with Diarrhea on Deployment in the Tropics: Experience from the European Union Training Mission (EUTM Mali

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    Hagen Frickmann

    2015-01-01

    Full Text Available Introduction. Since 2013, European soldiers have been deployed on the European Union Training Mission (EUTM in Mali. From the beginning, diarrhea has been among the most “urgent” concerns. Diarrhea surveillance based on deployable real-time PCR equipment was conducted between December 2013 and August 2014. Material and Methods. In total, 53 stool samples were obtained from 51 soldiers with acute diarrhea. Multiplex PCR panels comprised enteroinvasive bacteria, diarrhea-associated Escherichia coli (EPEC, ETEC, EAEC, and EIEC, enteropathogenic viruses, and protozoa. Noroviruses were characterized by sequencing. Cultural screening for Enterobacteriaceae with extended-spectrum beta-lactamases (ESBL with subsequent repetitive sequence-based PCR (rep-PCR typing was performed. Clinical information was assessed. Results. Positive PCR results for diarrhea-associated pathogens were detected in 43/53 samples, comprising EPEC (n=21, ETEC (n=19, EAEC (n=15, Norovirus (n=10, Shigella spp./EIEC (n=6, Cryptosporidium parvum (n=3, Giardia duodenalis (n=2, Salmonella spp. (n=1, Astrovirus (n=1, Rotavirus (n=1, and Sapovirus (n=1. ESBL-positive Enterobacteriaceae were grown from 13 out of 48 samples. Simultaneous infections with several enteropathogenic agents were observed in 23 instances. Symptoms were mild to moderate. There were hints of autochthonous transmission. Conclusions. Multiplex real-time PCR proved to be suitable for diarrhea surveillance on deployment. Etiological attribution is challenging in cases of detection of multiple pathogens.

  13. A Two-Tube Multiplex Reverse Transcription PCR Assay for Simultaneous Detection of Viral and Bacterial Pathogens of Infectious Diarrhea

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    Ji Wang

    2014-01-01

    Full Text Available Diarrhea caused by viral and bacterial infections is a major health problem in developing countries. The purpose of this study is to develop a two-tube multiplex PCR assay using automatic electrophoresis for simultaneous detection of 13 diarrhea-causative viruses or bacteria, with an intended application in provincial Centers for Diseases Control and Prevention, China. The assay was designed to detect rotavirus A, norovirus genogroups GI and GII, human astrovirus, enteric adenoviruses, and human bocavirus (tube 1, and Salmonella, Vibrio parahaemolyticus, diarrheagenic Escherichia coli, Campylobacter jejuni, Shigella, Yersinia, and Vibrio cholera (tube 2. The analytical specificity was examined with positive controls for each pathogen. The analytical sensitivity was evaluated by performing the assay on serial tenfold dilutions of in vitro transcribed RNA, recombinant plasmids, or bacterial culture. A total of 122 stool samples were tested by this two-tube assay and the results were compared with those obtained from reference methods. The two-tube assay achieved a sensitivity of 20–200 copies for a single virus and 102-103 CFU/mL for bacteria. The clinical performance demonstrated that the two-tube assay had comparable sensitivity and specificity to those of reference methods. In conclusion, the two-tube assay is a rapid, cost-effective, sensitive, specific, and high throughput method for the simultaneous detection of enteric bacteria and virus.

  14. [Viral contamination of food products: a poorly understood public health problem].

    Science.gov (United States)

    Ferrari, C K; Torres, E A

    1998-06-01

    Throughout the world there have been several epidemics of food-borne diseases (FBD) about which there is lack of sufficient information for public health institutions to take appropriate measures. This study was conducted for the purpose of contributing to the dissemination of information on these diseases and their etiologic agents, epidemiology, and control. The study was based on data from 61 sources, including review articles, reports of outbreaks, and databases. Results reveal considerable underregistration and lack of data on FBD throughout the various countries, with viruses being the second most important cause of FBD in the United States of America. Two agents, Norwalk virus and hepatitis A virus, were the fifth and sixth most frequent causes, respectively, although the former was the single most frequent cause of FBD in 1982 and the second most frequent cause of water-borne diseases during the period from 1986 to 1988. Despite the scarcity of information on the problem, rotavirus, poliovirus, hepatitis E virus, astrovirus, and small gastroenteric viruses are also important causes of FBD. We also discuss the importance of viral zoonoses, especially hemorrhagic fevers transmitted by contact with rodent feces and tick-borne viral encephalitides (Lassa fever). There is discussion of the controversial mad cow disease and its potential transmission through food products, as well as of dietary aspects of the management of AIDS and other viral infections. Finally, measures for the prevention and control of FBD are described.

  15. National laboratory surveillance of viral agents of gastroenteritis in Brazil.

    Science.gov (United States)

    Pereira, H G; Linhares, A C; Candeias, J A; Glass, R I

    1993-01-01

    In 1984 an enzyme immunoassay for rotavirus and adenovirus developed in the Virology Department of the Oswaldo Cruz Foundation was distributed to laboratories in 14 Brazilian states as part of a project to survey viral agents in fecal specimens from children with diarrhea. The ensuing surveillance continued for 3 to 5 years, and in 1991 the results obtained by all laboratories that tested 300 or more fecal specimens were reviewed at a workshop held in Belém. These results showed that rotavirus had been detected in 13% to 20% of the specimens tested, with positive specimens appearing to peak in the May to September "winter" or dry season in Brazil's central and southern states but not in the more tropical northern areas. Adenovirus, detected in 0.7% to 5.5% of the specimens tested for it, showed no seasonal variations. Many other viral agents known to cause gastroenteritis (e.g., astrovirus, small round-structured viruses, calcivirus, and group C rotavirus) were detected at centers that used electron microscopy. Picobirnavirus, a novel agent not yet associated with gastroenteritis in humans, was found by polyacrylamide gel electrophoresis in specimens from a number of centers using this technique. Vaccines to prevent rotavirus that are currently under development would be of great use in Brazil, where rotavirus is the most common cause of childhood diarrhea. Improved diagnostics will be required to assess the importance of the other viral agents.

  16. Novel divergent nidovirus in a python with pneumonia.

    Science.gov (United States)

    Bodewes, Rogier; Lempp, Charlotte; Schürch, Anita C; Habierski, Andre; Hahn, Kerstin; Lamers, Mart; von Dörnberg, Katja; Wohlsein, Peter; Drexler, Jan Felix; Haagmans, Bart L; Smits, Saskia L; Baumgärtner, Wolfgang; Osterhaus, Albert D M E

    2014-11-01

    The order Nidovirales contains large, enveloped viruses with a non-segmented positive-stranded RNA genome. Nidoviruses have been detected in man and various animal species, but, to date, there have been no reports of nidovirus in reptiles. In the present study, we describe the detection, characterization, phylogenetic analyses and disease association of a novel divergent nidovirus in the lung of an Indian python (Python molurus) with necrotizing pneumonia. Characterization of the partial genome (>33 000 nt) of this virus revealed several genetic features that are distinct from other nidoviruses, including a very large polyprotein 1a, a putative ribosomal frameshift signal that was identical to the frameshift signal of astroviruses and retroviruses and an accessory ORF that showed some similarity with the haemagglutinin-neuraminidase of paramyxoviruses. Analysis of genome organization and phylogenetic analysis of polyprotein 1ab suggests that this virus belongs to the subfamily Torovirinae. Results of this study provide novel insights into the genetic diversity within the order Nidovirales.

  17. The 12 Gastrointestinal Pathogens Spectrum of Acute Infectious Diarrhea in a Sentinel Hospital, Shenzhen, China

    Science.gov (United States)

    Shen, Hongwei; Zhang, Jinjin; Li, Yinghui; Xie, Sirou; Jiang, Yixiang; Wu, Yanjie; Ye, Yuhui; Yang, Hong; Mo, Haolian; Situ, Chaoman; Hu, Qinghua

    2016-01-01

    Acute infectious gastroenteritis is one of the most common diseases among all ages, particularly in developing countries. The pathogen spectrum may differ among different regions and seasons. To investigate the etiology of acute diarrhea in Shenzhen, a prospective study was conducted from August 2014 to September 2015. Stools from 412 patients with diarrhea (286 of whom were adults) including the general epidemiological information of the patients were collected. The 19 pathogens were detected by conventional culture method or multiplex PCR assay, which included five viruses (rotavirus, adenovirus, sapovirus, norovirus, and astrovirus), 11 bacterial pathogens (Salmonella, Campylobacter jejuni, Shigella, Listeria monocytogenes, Vibrio parahaemolyticus, Vibrio cholera, Enterohemorrhagic (EHEC), enteropathogenic (EPEC), enteroinvasive (EIEC), enterotoxigenic (ETEC); and enteroaggregative Escherichia coli (EAEC)) and three parasites (Entamoeba histolytica, Giardia lamblia, and Cryptosporidium parvum). A potential pathogen and coinfection was found in 41.5 and 7.0% of cases, respectively. The bacterial infection was the dominant cause of diarrhea (32.3%), and the three most frequently identified organisms were Salmonella (12.1%), ETEC (8.0%), and Campylobacter jejuni (4.9%). Salmonella enteritidis was the leading serotype of Salmonella sp. Norovirus (8.3%) and sapovirus (2.2%) were the most common viral pathogens, followed by adenovirus (1.5%) and rotavirus (1.2%). No EHEC, L. monocytogenes, V. cholera, Shigella, and parasites were found. The single most important causes of diarrhea were Salmonella spp. and Campylobacter jejuni, which points toward the need for testing and surveillance for these pathogens in this region. PMID:27965649

  18. The nineteen gastrointestinal pathogens spectrum of acute infectious diarrhea in a sentinel hospital, Shenzhen, China

    Directory of Open Access Journals (Sweden)

    Hongwei Shen

    2016-11-01

    Full Text Available Acute infectious gastroenteritis is one of the most common diseases among all ages, particularly in developing countries. The pathogen spectrum may differ among different regions and seasons. To investigate the etiology of acute diarrhea in Shenzhen, a prospective study was conducted from August 2014 to September 2015. Stools from 412 patients with diarrhea (286 of whom were adults including the general epidemiological information of the patients were collected. The 19 pathogens were detected by conventional culture method or multiplex PCR assay, which included five viruses (rotavirus, adenovirus, sapovirus, norovirus, and astrovirus,11 bacterial pathogens (Salmonella, Campylobacter jejuni, Shigella, Listeria monocytogenes, Vibrio parahaemolyticus, Vibrio cholera, Enterohemorrhagic (EHEC, enteropathogenic (EPEC, enteroinvasive (EIEC, enterotoxigenic (ETEC; and enteroaggregative Escherichia coli (EAEC and three parasites (Entamoeba histolytica, Giardia lamblia, and Cryptosporidium parvum. A potential pathogen and coinfection was found in 41.5% and 7.0% of cases, respectively. The bacterial infection was the dominant cause of diarrhea (32.3%, and the three most frequently identified organisms were Salmonella (12.1%, ETEC (8.0%, and Campylobacter jejuni (4.9%. Salmonella enteritidis was the leading serotype of Salmonella spp.. Norovirus (8.3% and sapovirus (2.2% were the most common viral pathogens, followed by adenovirus (1.5% and rotavirus (1.2%. The single most important causes of diarrhea were Salmonella spp. and Campylobacter jejuni, which points toward the need for testing and surveillance for these pathogens in this region.

  19. The 12 Gastrointestinal Pathogens Spectrum of Acute Infectious Diarrhea in a Sentinel Hospital, Shenzhen, China.

    Science.gov (United States)

    Shen, Hongwei; Zhang, Jinjin; Li, Yinghui; Xie, Sirou; Jiang, Yixiang; Wu, Yanjie; Ye, Yuhui; Yang, Hong; Mo, Haolian; Situ, Chaoman; Hu, Qinghua

    2016-01-01

    Acute infectious gastroenteritis is one of the most common diseases among all ages, particularly in developing countries. The pathogen spectrum may differ among different regions and seasons. To investigate the etiology of acute diarrhea in Shenzhen, a prospective study was conducted from August 2014 to September 2015. Stools from 412 patients with diarrhea (286 of whom were adults) including the general epidemiological information of the patients were collected. The 19 pathogens were detected by conventional culture method or multiplex PCR assay, which included five viruses (rotavirus, adenovirus, sapovirus, norovirus, and astrovirus), 11 bacterial pathogens (Salmonella, Campylobacter jejuni, Shigella, Listeria monocytogenes, Vibrio parahaemolyticus, Vibrio cholera, Enterohemorrhagic (EHEC), enteropathogenic (EPEC), enteroinvasive (EIEC), enterotoxigenic (ETEC); and enteroaggregative Escherichia coli (EAEC)) and three parasites (Entamoeba histolytica, Giardia lamblia, and Cryptosporidium parvum). A potential pathogen and coinfection was found in 41.5 and 7.0% of cases, respectively. The bacterial infection was the dominant cause of diarrhea (32.3%), and the three most frequently identified organisms were Salmonella (12.1%), ETEC (8.0%), and Campylobacter jejuni (4.9%). Salmonella enteritidis was the leading serotype of Salmonella sp. Norovirus (8.3%) and sapovirus (2.2%) were the most common viral pathogens, followed by adenovirus (1.5%) and rotavirus (1.2%). No EHEC, L. monocytogenes, V. cholera, Shigella, and parasites were found. The single most important causes of diarrhea were Salmonella spp. and Campylobacter jejuni, which points toward the need for testing and surveillance for these pathogens in this region.

  20. Avian nephritis virus (ANV) on Brazilian chickens farms: circulating genotypes and intra-genotypic diversity.

    Science.gov (United States)

    Espinoza, Luis Luna; Beserra, Laila A R; Soares, Rodrigo M; Gregori, Fabio

    2016-12-01

    Avian nephritis virus (ANV), which belongs to the family Astroviridae, is associated with different clinical manifestations (including enteric disorders). Despite being frequently found in the avian industry worldwide, information regarding genetic features of these viruses in Brazil is scarce. Therefore, sixty fecal sample pools (5-6 birds of the same flock), representing 60 poultry farms from six Brazilian States, were screened using an astrovirus-specific hemi-nested-PCR assay targeting the conserved ORF1b gene, followed by nucleotide sequencing of amplified products. PCR and phylogenetic analysis confirmed the detection of 21 positive samples to ANV (35 %). In order to investigate the genetic diversity represented by these viruses, amplification, cloning and phylogenetic analysis of the deduced amino acid sequence of ORF2 gene were attempted. Eight samples were successfully cloned (generating 32 clones in total) and sequenced. Based on phylogenetic analysis of ORF2, sequences defined in this study were classified into three genotypes: genotype 5, which has already been described in birds, and two other novel genotypes, tentatively named genotype 8 and 9, all of which occurred in single or mixed infections. Moreover, high intra-genotypic diversity and co-circulation of distinct strains in a same host population were observed. This study revealed the presence of new strains of ANV in Brazilian poultry and their circulation in commercial chicken flocks.

  1. Development of the Intestinal RNA Virus Community of Healthy Broiler Chickens.

    Directory of Open Access Journals (Sweden)

    Jigna D Shah

    Full Text Available Several RNA viruses such as astrovirus, rotavirus, reovirus and parvovirus have been detected in both healthy and diseased commercial poultry flocks. The aim of this study was to characterize (a the development of the RNA viral community in the small intestines of healthy broiler chickens from hatch through 6 weeks of age (market age and (b the contribution of the breeder source vs. bird age in development of the community structure. Intestinal tissue samples were harvested from breeders and their progeny, processed for viral RNA extraction and sequenced using Illumina Hiseq sequencing technology resulting in 100 bp PE reads. The results from this study indicated that the breeder source influenced the RNA viral community only at hatch but later environment i.e. bird age had the more significant effect. The most abundant RNA viral family detected at 2, 4 and 6 weeks of age was Astroviridae, which decreased in abundance with age while the abundance of Picornaviridae increased with age.

  2. New viruses in veterinary medicine, detected by metagenomic approaches.

    Science.gov (United States)

    Belák, Sándor; Karlsson, Oskar E; Blomström, Anne-Lie; Berg, Mikael; Granberg, Fredrik

    2013-07-26

    In our world, which is faced today with exceptional environmental changes and dramatically intensifying globalisation, we are encountering challenges due to many new factors, including the emergence or re-emergence of novel, so far "unknown" infectious diseases. Although a broad arsenal of diagnostic methods is at our disposal, the majority of the conventional diagnostic tests is highly virus-specific or is targeted entirely towards a limited group of infectious agents. This specificity complicates or even hinders the detection of new or unexpected pathogens, such as new, emerging or re-emerging viruses or novel viral variants. The recently developed approaches of viral metagenomics provide an effective novel way to screen samples and detect viruses without previous knowledge of the infectious agent, thereby enabling a better diagnosis and disease control, in line with the "One World, One Health" principles (www.oneworldonehealth.org). Using metagenomic approaches, we have recently identified a broad variety of new viruses, such as novel bocaviruses, Torque Teno viruses, astroviruses, rotaviruses and kobuviruses in porcine disease syndromes, new virus variants in honeybee populations, as well as a range of other infectious agents in further host species. These findings indicate that the metagenomic detection of viral pathogens is becoming now a powerful, cultivation-independent, and useful novel diagnostic tool in veterinary diagnostic virology.

  3. Emergence of Enteric Viruses in Production Chickens Is a Concern for Avian Health

    Directory of Open Access Journals (Sweden)

    Elena Mettifogo

    2014-01-01

    Full Text Available Several viruses have been identified in recent years in the intestinal contents of chickens and turkeys with enteric problems, which have been observed in commercial farms worldwide, including Brazil. Molecular detection of these viruses in Brazil can transform to a big threat for poultry production due to risk for intestinal integrity. This disease is characterized by severely delayed growth, low uniformity, lethargy, watery diarrhea, delayed feed consumption, and a decreased conversion rate. Chicken astrovirus (CAstV, rotavirus, reovirus, chicken parvovirus (ChPV, fowl adenovirus of subgroup I (FAdV-1, and avian nephritis virus (ANV were investigated using the conventional polymerase chain reaction (PCR and the reverse transcription polymerase chain reaction (RT-PCR. In addition, the infectious bronchitis virus (IBV, which may play a role in enteric disease, was included. The viruses most frequently detected, either alone or in concomitance with other viruses, were IBV, ANV, rotavirus, and CAstV followed by parvovirus, reovirus, and adenovirus. This study demonstrates the diversity of viruses in Brazilian chicken flocks presenting enteric problems characterized by diarrhea, growth retard, loss weight, and mortality, which reflects the multicausal etiology of this disease.

  4. Detection of enteric pathogens in Turkey flocks affected with severe enteritis, in Brazil.

    Science.gov (United States)

    Moura-Alvarez, Joelma; Nuñez, Luis F N; Astolfi-Ferreira, Claudete S; Knöbl, Terezinha; Chacón, Jorge L; Moreno, Andrea M; Jones, Richard C; Ferreira, Antonio J Piantino

    2014-08-01

    Twenty-two flocks of turkeys affected by enteric problems, with ages between 10 and 104 days and located in the Southern region of Brazil, were surveyed for turkey by PCR for turkey astrovirus type 2 (TAstV-2), turkey coronavirus (TCoV), hemorrhagic enteritis virus (HEV), rotavirus, reovirus, Salmonella spp., and Lawsonia intracellularis (Li) infections. Eleven profiles of pathogen combination were observed. The most frequently encountered pathogen combinations were TCoV-Li, followed by TCoV-TAstV-2-Li, TCoV-TastV-2. Only TCoV was detected as the sole pathogen in three flocks. Eight and 19 flocks of the 22 were positive for TAstV-2 and TCoV, respectively. Six were positive for Salmonella spp. and L. intracellularis was detected in 12 turkey flocks. Reovirus and HEV were not detected in this survey. These results throw new light on the multiple etiology of enteritis in turkeys. The implications of these findings and their correlation with the clinical signs are comprehensively discussed, illustrating the complexity of the enteric diseases.

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

    Science.gov (United States)

    Portes, Silvana Augusta Rodrigues; Volotão, Eduardo de Mello; Rocha, Monica Simões; Rebelo, Maria Cristina; Xavier, Maria da Penha Trindade Pinheiro; de Assis, Rosane Maria; Rose, Tatiana Lundgren; Miagostovich, Marize Pereira; Leite, José Paulo Gagliardi; Carvalho-Costa, Filipe Anibal

    2016-01-01

    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. PMID:27223654

  6. Molecular characterization of viruses associated with gastrointestinal infection in HIV-positive patients

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    Raquel C Silva

    2010-12-01

    Full Text Available BACKGROUND: Diarrhea is a major cause of morbidity and mortality among HIV-infected patients worldwide. OBJECTIVE: We sought to determine the frequency of viral gastrointestinal infections among Brazilian HIV-infected patients with diarrhea. METHODS: A collection of 90 fecal specimens from HIV-infected individuals with diarrhea, previously tested for the presence of bacteria and parasite was analyzed by polymerase chain reaction and sequence analysis for the presence of enteric viruses such as astrovirus, norovirus, rotavirus groups A, B and C, adenovirus, herpes simplex virus, Epstein-Barr virus, cytomegalovirus, and human bocavirus. RESULTS: Twenty patients (22.2%; n = 90 were infected with parasites (11 single infections and nine coinfected with virus. Enteropathogenic bacteria were not found. Virus infections were detected in 28.9% (26/90 of the specimens. Cytomegalovirus was the most common virus detected (24.4%; 22/90. Coinfections with viruses and/or parasite were observed in 10 (11.1% samples. CONCLUSION: Gastrointestinal virus infections were more frequent than parasitic or bacterial infections in this patient population.

  7. Viral metagenomics analysis demonstrates the diversity of viral flora in piglet diarrhoeic faeces in China.

    Science.gov (United States)

    Zhang, Bin; Tang, Cheng; Yue, Hua; Ren, Yupeng; Song, Zhigang

    2014-07-01

    To investigate the diversity of viral flora, we used metagenomics to study the viral communities in a pooled faecal sample of 27 diarrhoeic piglets from intensive commercial farms in China. The 15 distinct mammalian viruses identified in the pooled diarrhoeic sample were, in order of abundance of nucleic acid sequence, Porcine epidemic diarrhea virus (PEDV), sapovirus, porcine bocavirus-4 (PBoV-4), sapelovirus, torovirus, coronavirus, PBoV-2, stool-associated single-stranded DNA virus (poSCV), astrovirus (AstV), kobuvirus, posavirus-1, porcine enterovirus-9 (PEV-9), porcine circovirus-like (po-circo-like) virus, picobirnavirus (PBV) and Torque teno sus virus 2 (TTSuV-2). The prevalence rate of each virus was verified from diarrhoeic and healthy piglets by PCR assay. A mean of 5.5 different viruses were shed in diarrhoeic piglets, and one piglet was in fact co-infected with 11 different viruses. By contrast, healthy piglets shed a mean of 3.2 different viruses. Compared with samples from healthy piglets, the co-infection of PEDV and PBoV had a high prevalence rate in diarrhoea samples, suggesting a correlation with the appearance of diarrhoea in piglets. Furthermore, we report here for the first time the presence of several recently described viruses in China, and the identification of novel genotypes. Therefore, our investigation results provide an unbiased survey of viral communities and prevalence in faecal samples of piglets.

  8. 天津市病毒性腹泻的病原学分析%Pathogenic Analysis on Viral Diarrhea in Tianjin

    Institute of Scientific and Technical Information of China (English)

    田宏; 雷玥; 刘杨; 张颖; 李佳萌

    2011-01-01

    目的 了解天津市病毒性腹泻的病原学分布情况.方法 于2009年3-10月从天津市儿童医院和宝坻医院收集感染性腹泻患者的粪便标本,采用酶联免疫技术(ELISA)检测轮状病毒,采用逆转录-聚合酶链反应(RT-PCR)方法检测杯状病毒、星状病毒,并对轮状病毒ELISA检测阳性标本进行分型.采用聚合酶链反应(PCR)检测腺病毒.结果 从天津市儿童医院收集儿童粪便标本124份,四种病毒检测阳性率顺位依次为杯状病毒(17.7%)>腺病毒(16.9%)>轮状病毒(12.1%)>星状病毒(1.6%),总检出率为48.4%.从宝坻医院收集成人粪便标本102份,四种病毒检测阳性率顺位依次为杯状病毒(8.8%)>腺病毒(3.9%)>轮状病毒(1.0%)>星状病毒(1.0%),总检出率为14.7%.16株轮状病毒G血清型阳性8株,G1为5株(31.2%),G3为3株(18.8%),8株未能分型(50%);P基因型分型结果阳性7株,P[4]为2株(12.5%),P[6]为1株(6.2%),P[8]为4株(25%),9株未能分型(56.3%).G型与P型组合P[4]G1为1株(6.2%),P[8]G3为1株(6.2%),P[8]G1为3株(18.8%).31株诺如病毒全部为GⅡ型.腺病毒主要是Ad40和Ad41型.星状病毒是1型.结论 本次调查的该市儿童病毒性腹泻发病率明显高于成年人,病毒分布呈现出多样性.%Objective To investigate the main pathogens of viral diarrhea in Tianjin. Methods The feces specimens of infectious diarrhea patients were collected from Tianjin Children's Hospital and Baodi Hospital. Rotavirus was detected by ELISA,calicivirus and astrovirus were detected by RT-PCR. Further strain characterization of rotavirus was carried out with RT-PCR and adenovirus were detected by PCR. Results Tested feces specimens with amount of 124 were collected from Tianjin Children's Hospital. Calicivirus was detected in 17.7% of the cases, adenoviruses in 16.9% , rotavirus in 12.1% and astrovirus in 1.6 %.The total positive rate was 48.4%. Tested feces specimens with amount of 102 were collected from Baodi

  9. The epidemiology of acute viral gastroenteritis in hospitalized children in Cordoba city, Argentina: an insight of disease burden Epidemiologia da gastrenterite viral aguda em crianças hospitalizadas na cidade de Córdoba, Argentina: uma percepção ao impacto da doença

    Directory of Open Access Journals (Sweden)

    Miguel O. GIORDANO

    2001-08-01

    Full Text Available Information concerning the disease burden of viral gastroenteritis has important implications for the use and monitoring the impact of public health policies. The present study, carried out in Córdoba city, Argentina, documents the epidemiology of severe viral diarrhea as well as the burden of viral gastrointestinal disease in the hospital children admission. A total of 133 stools were collected from hospitalized children (Town Childhood Hospital suffering from acute diarrhea and studied for the presence of Group A rotavirus, astrovirus and adenovirus 40/41 by enzyme-immuno assay, between November 1997 and October 1998. Enteric viruses accounted for 42.1% of the total diarrheal cases analyzed. Group A rotaviruses, astroviruses, adenoviruses 40/41 and mixed infections were found in 35.3, 4.5, 1.5, and 0.8% studied specimens respectively. We estimated that 1 in 27 children in the 0-35 month-old cohort/range would be annually hospitalized for a viral gastroenteritis illness. The major impact on viral diarrhea lies on rotaviral infection, accouting for 84.0% of the viral diarrheal cases analyzed and for approximately one third of severe diarrheas requiring hospital admission in Córdoba City, Argentina.Dados a respeito do impacto das gastrenterites virais têm implicações importantes para o monitoraramento do impacto das políticas de saúde pública. O presente estudo, feito na cidade de Córdoba, Argentina, documenta a epidemiologia da diarréia viral severa como também o impacto que a mesma representa na admissão hospitalar de crianças. Pesquisou-se a presença de Rotavírus grupo A, Astrovírus e Adenovírus 40/41 por ensaio imuno-enzimático em 133 amostras de fezes obtidas durante o período de novembro de 1997 a outubro de 1998, de crianças hospitalizadas por diarréia aguda. Os virus entéricos constituíram 42,1% do total de casos de diarréia analisados. Encontraram-se Rotavírus Grupo A, Astrovírus, Adenovírus 40/41 e infec

  10. The sensitivity and specificity of two PCR assays for detecting rabies virus%两种PCR方法检测狂犬病毒的敏感性和特异性比较

    Institute of Scientific and Technical Information of China (English)

    马莉珍; 柯雪梅; 肖建鹏; 陈清

    2011-01-01

    目的 比较Nested-PCR和SYBR Green Ⅰ Real-Time PCR两种方法检测狂犬病毒的敏感性、特异性和时效性.方法 对10倍连续稀释的狂犬病毒(疫苗株)核酸样品,采用Nested-PCR和SYBR Green Ⅰ Real-Time PCR方法进行平行检测,比较两种方法的敏感性:同时以登革热2型病毒、诺如病毒、星状病毒、EV71和乙型脑炎病毒核酸对两种方法的特异性进行评价.结果 SYBR Green Ⅰ Real-Time PCR方法可检测出2.3×106 copies/μl核酸分子,与NestedPCR方法相比,敏感度提高10倍.两种检测方法的特异性均好,与登革热2型病毒、诺如病毒、星状病毒、EV71和乙型脑炎病毒核酸均无交叉反应.SYBR Green Ⅰ Real-Time PCR方法检测花费3 h,检测时间比Nested-PCR方法节省2 h.结论 与Nested-PCR相比,SYBR Green Ⅰ Real-Time PCR是相对高效的检测狂犬病毒的方法.%Objective The aim of the study was to compare the sensitivity, specificity and time-consuming of Nested-PCR and SYBR Green I Real-Time PCR assays for detecting rabies virus (RABV). Methods A series of 10-fold dilutions of a RABV (vaccine strain) DNA was used for detection of the sensitivity of the Nested-PCR and SYBR Green I Real-Time PCR assays. EV71, Dengue type 2 virus, Norovirus, Astrovirus and Japanese encephalitis virus (JEV) were used to test the specificity of the assays. Results Sensitivity of SYBR Green I Real-Time PCR assay was 2.3×106 copies/μl which were 10 times more sensitive than that of Nested-PCR assay. There was no crossing-reaction with Dengue type 2 virus, Astrovirus,Norovirus, EV71 and JEV. The time-consuming of the SYBR Green I real-time PCR amplification was about 3 hours, 2 hours shorter than Nested-PCR. Conclusion For detection of Rabies Virus, SYBR Green I Real-Time PCR is a more sensitive and rapid assay compared with Nested-PCR.

  11. Molecular epidemiological survey on children with viral diarrhea in Tangshan%唐山地区儿童病毒性腹泻分子流行病学调查

    Institute of Scientific and Technical Information of China (English)

    余亮科; 赵文喆; 高捷; 张双; 张志坤

    2011-01-01

    Objective: To explore the current situation of diarrhea in children in Tangshan region from January 2007 to December 2010, and a epidemiological investigation and analysis on the viruses of diarrhea was conducted. Methods: 1 100 children with diarrhea were selected randomly from Tangshan region from January 2007 to December 2010, all the children received stool examination, a variety of methods were used to analyze the common pathogens and peak time of onset Results: Among 1 100 children, 582 children were found with rolavirus, accounting for 52. 91 % ; 258 children were found with norovirus, accounting for 23. 45% ; 89 children were found with sapovirus, accounting for 8. 09% ; 52 children were found with astrovirus, accounting for 4. 73 % ; 119 children were found with adenovirus, accounting for 10. 82%. The peak time of onset of rotavirus was October - December and January, while the peak time of onset of norovirus was July ~ September, the other viruses could be detected in each time period Conclusion: The peak age of onset among the children with diarrhea is below 5 years. The major viruses of diarrhea are rotavirus and norovirus; astrovirus, sapovirus and adenovirus are also common viruses. Different viruses have different peak times of onset, for rotavirus, the peak times of onset are autumn and winter; while for norovirus, the peak times of onset are summer and autumn; the other viruses have no obvious seasonal distribution.%目的:探讨唐山地区2007年1月~2010年12月儿童腹泻发病现状,并对腹泻病毒进行流行病学调查分析.方法:随机抽取唐山地区2007年1月~2010年12月腹泻儿童1 100例资料.所有患儿均经过粪便检查,采用多种方法分析常见病原菌及好发时间.结果:1 100例儿童中,轮状病毒582例,占52.91%;诺如病毒258例,占23.45%;札如病毒89例,占8.09%;星状病毒52例,占4.73%;腺病毒119例,占10.82%.其中轮状病毒在10~12月及1月高发,诺如病毒为7~9月高

  12. Is There Still Room for Novel Viral Pathogens in Pediatric Respiratory Tract Infections?

    Science.gov (United States)

    Taboada, Blanca; Espinoza, Marco A.; Isa, Pavel; Aponte, Fernando E.; Arias-Ortiz, María A.; Monge-Martínez, Jesús; Rodríguez-Vázquez, Rubén; Díaz-Hernández, Fidel; Zárate-Vidal, Fernando; Wong-Chew, Rosa María; Firo-Reyes, Verónica; del Río-Almendárez, Carlos N.; Gaitán-Meza, Jesús; Villaseñor-Sierra, Alberto; Martínez-Aguilar, Gerardo; Salas-Mier, Ma. del Carmen; Noyola, Daniel E.; Pérez-Gónzalez, Luis F.; López, Susana; Santos-Preciado, José I.; Arias, Carlos F.

    2014-01-01

    Viruses are the most frequent cause of respiratory disease in children. However, despite the advanced diagnostic methods currently in use, in 20 to 50% of respiratory samples a specific pathogen cannot be detected. In this work, we used a metagenomic approach and deep sequencing to examine respiratory samples from children with lower and upper respiratory tract infections that had been previously found negative for 6 bacteria and 15 respiratory viruses by PCR. Nasal washings from 25 children (out of 250) hospitalized with a diagnosis of pneumonia and nasopharyngeal swabs from 46 outpatient children (out of 526) were studied. DNA reads for at least one virus commonly associated to respiratory infections was found in 20 of 25 hospitalized patients, while reads for pathogenic respiratory bacteria were detected in the remaining 5 children. For outpatients, all the samples were pooled into 25 DNA libraries for sequencing. In this case, in 22 of the 25 sequenced libraries at least one respiratory virus was identified, while in all other, but one, pathogenic bacteria were detected. In both patient groups reads for respiratory syncytial virus, coronavirus-OC43, and rhinovirus were identified. In addition, viruses less frequently associated to respiratory infections were also found. Saffold virus was detected in outpatient but not in hospitalized children. Anellovirus, rotavirus, and astrovirus, as well as several animal and plant viruses were detected in both groups. No novel viruses were identified. Adding up the deep sequencing results to the PCR data, 79.2% of 250 hospitalized and 76.6% of 526 ambulatory patients were positive for viruses, and all other children, but one, had pathogenic respiratory bacteria identified. These results suggest that at least in the type of populations studied and with the sampling methods used the odds of finding novel, clinically relevant viruses, in pediatric respiratory infections are low. PMID:25412469

  13. Diagnosis of viral gastroenteritis in children: interpretation of real-time PCR results and relation to clinical symptoms.

    Science.gov (United States)

    Corcoran, M S; van Well, G T J; van Loo, I H M

    2014-10-01

    Molecular methods such as real-time polymerase chain reaction (PCR) are rapidly replacing traditional tests to detect fecal viral pathogens in childhood diarrhea. This technique has now increased the analytical sensitivity so drastically that positive results are found in asymptomatic children, leading to complex interpretation of real-time PCR results and difficult distinction between asymptomatic shedding and etiological cause of disease. We performed a review of the literature including pediatric studies using real-time PCR and a minimal inclusion period of one year to exclude bias by seasonality. We searched for studies on rotavirus, norovirus, adenovirus, astrovirus, and sapovirus, known to be the most common viruses to cause gastroenteritis in the pediatric population. For these viruses, we summarized the detection rates in hospitalized and community-based children with clinical symptoms of gastroenteritis, as well as subjects with asymptomatic viral shedding. Moreover, insight is given into the different viral sero- and genotypes causing pediatric gastroenteritis. We also discuss the scoring systems for severity of disease and their clinical value. A few published proposals have been made to improve the clinical interpretation of real-time PCR results, which we recapitulate and discuss in this review. We propose using the semi-quantitative measure of real-time PCR, as a surrogate for viral load, in relation to the severity score to distinguish asymptomatic viral shedding from clinically relevant disease. Overall, this review provides a better understanding of the scope of childhood gastroenteritis, discusses a method to enhance the interpretation of real-time PCR results, and proposes conditions for future research to enhance clinical implementation.

  14. Is there still room for novel viral pathogens in pediatric respiratory tract infections?

    Directory of Open Access Journals (Sweden)

    Blanca Taboada

    Full Text Available Viruses are the most frequent cause of respiratory disease in children. However, despite the advanced diagnostic methods currently in use, in 20 to 50% of respiratory samples a specific pathogen cannot be detected. In this work, we used a metagenomic approach and deep sequencing to examine respiratory samples from children with lower and upper respiratory tract infections that had been previously found negative for 6 bacteria and 15 respiratory viruses by PCR. Nasal washings from 25 children (out of 250 hospitalized with a diagnosis of pneumonia and nasopharyngeal swabs from 46 outpatient children (out of 526 were studied. DNA reads for at least one virus commonly associated to respiratory infections was found in 20 of 25 hospitalized patients, while reads for pathogenic respiratory bacteria were detected in the remaining 5 children. For outpatients, all the samples were pooled into 25 DNA libraries for sequencing. In this case, in 22 of the 25 sequenced libraries at least one respiratory virus was identified, while in all other, but one, pathogenic bacteria were detected. In both patient groups reads for respiratory syncytial virus, coronavirus-OC43, and rhinovirus were identified. In addition, viruses less frequently associated to respiratory infections were also found. Saffold virus was detected in outpatient but not in hospitalized children. Anellovirus, rotavirus, and astrovirus, as well as several animal and plant viruses were detected in both groups. No novel viruses were identified. Adding up the deep sequencing results to the PCR data, 79.2% of 250 hospitalized and 76.6% of 526 ambulatory patients were positive for viruses, and all other children, but one, had pathogenic respiratory bacteria identified. These results suggest that at least in the type of populations studied and with the sampling methods used the odds of finding novel, clinically relevant viruses, in pediatric respiratory infections are low.

  15. Characterization of a genetically heterogeneous porcine rotavirus C, and other viruses present in the fecal virome of a non-diarrheic Belgian piglet.

    Science.gov (United States)

    Theuns, Sebastiaan; Conceição-Neto, Nádia; Zeller, Mark; Heylen, Elisabeth; Roukaerts, Inge D M; Desmarets, Lowiese M B; Van Ranst, Marc; Nauwynck, Hans J; Matthijnssens, Jelle

    2016-09-01

    Next-generation sequencing (NGS) technologies are becoming increasingly accessible, leading to an expanded interest in the composition of the porcine enteric virome. In the present study, the fecal virome of a non-diarrheic Belgian piglet was determined. Although the virome of only a single piglet was analyzed, some interesting data were obtained, including the second complete genome of a pig group C rotavirus (RVC). This Belgian strain was only distantly related to the only other completely characterized pig RVC strain, Cowden. Its relatedness to RVC strains from other host species was also analyzed and the porcine strain found in our study was only distantly related to RVCs detected in humans and cows. The gene encoding the outer capsid protein VP7 belonged to the rare porcine G3 genotype, which might be serologically distinct from most other pig RVC strains. A putative novel RVC VP6 genotype was identified as well. A group A rotavirus strain also present in this fecal sample contained the rare pig genotype combination G11P[27], but was only partially characterized. Typical pig RVA genotypes I5, A8, and T7 were found for the viral proteins VP6, NSP1, and NSP3, respectively. Interestingly, the fecal virome of the piglet also contained an astrovirus and an enterovirus, of which the complete genomes were characterized. Results of the current study indicate that many viruses may be present simultaneously in fecal samples of non-diarrheic piglets. In this study, these viruses could not be directly associated with any disease, but still they might have had a potential subclinical impact on pig growth performance. The fast evolution of NGS will be a powerful tool for future diagnostics in veterinary practice. Its application will certainly lead to better insights into the relevance of many (sub)clinical enteric viral infections, that may have remained unnoticed using traditional diagnostic techniques. This will stimulate the development of new and durable prophylactic

  16. Molecular epidemiology of enteric viruses in patients with acute gastroenteritis in Aichi prefecture, Japan, 2008/09-2013/14.

    Science.gov (United States)

    Nakamura, Noriko; Kobayashi, Shinichi; Minagawa, Hiroko; Matsushita, Tadashi; Sugiura, Wataru; Iwatani, Yasumasa

    2016-07-01

    Acute gastroenteritis is a critical infectious disease that affects infants and young children throughout the world, including Japan. This retrospective study was conducted from September 2008 to August 2014 (six seasons: 2008/09-2013/14) to investigate the incidence of enteric viruses responsible for 1,871 cases of acute gastroenteritis in Aichi prefecture, Japan. Of the 1,871 cases, 1,100 enteric viruses were detected in 978 samples, of which strains from norovirus (NoV) genogroup II (60.9%) were the most commonly detected, followed by strains of rotavirus A (RVA) (23.2%), adenovirus (AdV) type 41 (8.2%), sapovirus (SaV) (3.6%), human astrovirus (HAstV) (2.8%), and NoV genogroup I (1.3%). Sequencing of the NoV genogroup II (GII) strains revealed that GII.4 was the most common genotype, although four different GII.4 variants were also identified. The most common G-genotype of RVA was G1 (63.9%), followed by G3 (27.1%), G2 (4.7%) and G9 (4.3%). Three genogroups of SaV strains were found: GI (80.0%), GII (15.0%), and GV (5.0%). HAstV strains were genotyped as HAstV-1 (80.6%), HAstV-8 (16.1%), and HAstV-3 (3.2%). These results show that NoV GII was the leading cause of sporadic acute viral gastroenteritis, although a variety of enteric viruses were detected during the six-season surveillance period.

  17. Factors associated with usage of antimicrobials in commercial mink (Neovison vison) production in Denmark.

    Science.gov (United States)

    Jensen, V F; Sommer, H M; Struve, T; Clausen, J; Chriél, M

    2016-04-01

    The American mink (Neovison vison) is used for commercial fur production in Denmark. In recent years, antimicrobial prescription for Danish mink has been increasing. In this study, the patterns and trends in antimicrobial use in mink were described and a multi-variable variance analysis was carried out with the objective of identifying risk factors for antimicrobial use on herd level. The study was based on register data for 2007-2012. Information on antimicrobial use was obtained from the national database VetStat, monitoring all medicinal products used for animals on prescription level. Data on microbiological feed quality was obtained from the Voluntary Feed Control under the Mink producers Organization, and data on herd size and the relation between farm and feed producer was obtained from the registers at Kopenhagen Fur, based on yearly reporting from the mink producers. Descriptive analysis showed a clear significant effect of season on antimicrobial use, with a peak in "treatment proportions", TP (defined daily doses per kg biomass-days) in May, around the time of whelping, and a high level in the following months. In autumn, a minor peak in antimicrobial use occurred throughout the study period. From 2007 to 2011, a 102% increase in annual antimicrobial TP was noted; on herd level, the increase was associated with an increasing frequency of prescription, and a decrease in the amounts prescribed in months with prescription. A binomial model showed that on herd level, the annual number of months with antimicrobial prescription was significantly (pprescription patterns varied significantly between veterinarians, and some veterinarians were associated with both larger and more frequent prescriptions of antimicrobials at herd level. Herd size is associated with different prescription patterns. Finally, infection with Pseudomonas aeruginosa, astrovirus, influenza virus and Salmonella spp. was associated with an increase in antimicrobial use.

  18. ENTEROPATHOGENS DETECTED IN A DAYCARE CENTER, SOUTHEASTERN BRAZIL: BACTERIA, VIRUS, AND PARASITE RESEARCH

    Science.gov (United States)

    Castro, Edna Donizetti Rossi; Germini, Marcela Cristina Braga Yassaka; Mascarenhas, Joana D'Arc Pereira; Gabbay, Yvone Benchimol; de Lima, Ian Carlos Gomes; Lobo, Patrícia dos Santos; Fraga, Valéria Daltibari; Conceição, Luciana Moran; Machado, Ricardo Luiz Dantas; Rossit, Andréa Regina Baptista

    2015-01-01

    Introduction: The objective of this study was to determine the prevalence and etiological profile of enteropathogens in children from a daycare center. Methods: From October 2010 to February 2011 stool samples from 100 children enrolled in a government daycare center in the municipality of São José do Rio Preto, in the state of São Paulo, were collected and analyzed. Results: A total of 246 bacteria were isolated in 99% of the fecal samples; 129 were in the diarrheal group and 117 in the non-diarrheal group. Seventy-three strains of Escherichia coli were isolated, 19 of Enterobacter, one of Alcaligenes and one of Proteus. There were 14 cases of mixed colonization with Enterobacter and E. coli. Norovirus and Astrovirus were detected in children with clinical signs suggestive of diarrhea. These viruses were detected exclusively among children residing in urban areas. All fecal samples were negative for the presence of the rotavirus species A and C. The presence of Giardia lamblia, Entamoeba coli, Endolimax nana and hookworm was observed. A significant association was found between food consumption outside home and daycare center and the presence of intestinal parasites. Conclusions: For children of this daycare center, intestinal infection due to pathogens does not seem to have contributed to the occurrence of diarrhea or other intestinal symptoms. The observed differences may be due to the wide diversity of geographical, social and economic characteristics and the climate of Brazil, all of which have been reported as critical factors in the modulation of the frequency of different enteropathogens. PMID:25651323

  19. Removal of human pathogenic viruses in a down-flow hanging sponge (DHS) reactor treating municipal wastewater and health risks associated with utilization of the effluent for agricultural irrigation.

    Science.gov (United States)

    Kobayashi, Naohiro; Oshiki, Mamoru; Ito, Toshihiro; Segawa, Takahiro; Hatamoto, Masashi; Kato, Tsuyoshi; Yamaguchi, Takashi; Kubota, Kengo; Takahashi, Masanobu; Iguchi, Akinori; Tagawa, Tadashi; Okubo, Tsutomu; Uemura, Shigeki; Harada, Hideki; Motoyama, Toshiki; Araki, Nobuo; Sano, Daisuke

    2017-03-01

    A down-flow hanging sponge (DHS) reactor has been developed as a cost-effective wastewater treatment system that is adaptable to local conditions in low-income countries. A pilot-scale DHS reactor previously demonstrated stable reduction efficiencies for chemical oxygen demand (COD) and ammonium nitrogen over a year at ambient temperature, but the pathogen reduction efficiency of the DHS reactor has yet to be investigated. In the present study, the reduction efficiency of a pilot-scale DHS reactor fed with municipal wastewater was investigated for 10 types of human pathogenic viruses (norovirus GI, GII and GIV, aichivirus, astrovirus, enterovirus, hepatitis A and E viruses, rotavirus, and sapovirus). DHS influent and effluent were collected weekly or biweekly for 337 days, and concentrations of viral genomes were determined by microfluidic quantitative PCR. Aichivirus, norovirus GI and GII, enterovirus, and sapovirus were frequently detected in DHS influent, and the log10 reduction (LR) of these viruses ranged from 1.5 to 3.7. The LR values for aichivirus and norovirus GII were also calculated using a Bayesian estimation model, and the average LR (±standard deviation) values for aichivirus and norovirus GII were estimated to be 1.4 (±1.5) and 1.8 (±2.5), respectively. Quantitative microbial risk assessment was conducted to calculate a threshold reduction level for norovirus GII that would be required for the use of DHS effluent for agricultural irrigation, and it was found that LRs of 2.6 and 3.7 for norovirus GII in the DHS effluent were required in order to not exceed the tolerable burden of disease at 10(-4) and 10(-6) disability-adjusted life years loss per person per year, respectively, for 95% of the exposed population during wastewater reuse for irrigation.

  20. Cosavirus, Salivirus and Bufavirus in Diarrheal Tunisian Infants

    Science.gov (United States)

    Ayouni, Siwar; Estienney, Marie; Hammami, Sabeur; Neji Guediche, Mohamed; Pothier, Pierre; Aouni, Mahjoub; Belliot, Gael

    2016-01-01

    Three newly discovered viruses have been recently described in diarrheal patients: Cosavirus (CosV) and Salivirus (SalV), two picornaviruses, and Bufavirus (BuV), a parvovirus. The detection rate and the role of these viruses remain to be established in acute gastroenteritis (AGE) in diarrheal Tunisian infants. From October 2010 through March 2012, stool samples were collected from 203 children <5 years-old suffering from AGE and attending the Children’s Hospital in Monastir, Tunisia. All samples were screened for CosV, SalV and BuV as well as for norovirus (NoV) and group A rotavirus (RVA) by molecular biology. Positive samples for the three screened viruses were also tested for astrovirus, sapovirus, adenovirus, and Aichi virus, then genotyped when technically feasible. During the study period, 11 (5.4%) samples were positive for one of the three investigated viruses: 2 (1.0%) CosV-A10, 7 (3.5%) SalV-A1 and 2 (1.0%) BuV-1, whereas 71 (35.0%) children were infected with NoV and 50 (24.6%) with RVA. No mixed infections involving the three viruses were found, but multiple infections with up to 4 classic enteric viruses were found in all cases. Although these viruses are suspected to be responsible for AGE in children, our data showed that this association was uncertain since all infected children also presented infections with several enteric viruses, suggesting here potential water-borne transmission. Therefore, further studies with large cohorts of healthy and diarrheal children will be needed to evaluate their clinical role in AGE. PMID:27631733

  1. Avian rotavirus enteritis - an updated review.

    Science.gov (United States)

    Dhama, Kuldeep; Saminathan, Mani; Karthik, Kumaragurubaran; Tiwari, Ruchi; Shabbir, Muhammad Zubair; Kumar, Naveen; Malik, Yashpal Singh; Singh, Raj Kumar

    2015-01-01

    Rotaviruses (RVs) are among the leading causes of enteritis and diarrhea in a number of mammalian and avian species, and impose colossal loss to livestock and poultry industry globally. Subsequent to detection of rotavirus in mammalian hosts in 1973, avian rotavirus (AvRV) was first reported in turkey poults in USA during 1977 and since then RVs of group A (RVA), D (RVD), F (RVF) and G (RVG) have been identified around the globe. Besides RVA, other AvRV groups (RVD, RVF and RVG) may also contribute to disease. However, their significance has yet to be unraveled. Under field conditions, co-infection of AvRVs occurs with other infectious agents such as astroviruses, enteroviruses, reoviruses, paramyxovirus, adenovirus, Salmonella, Escherichia coli, cryptosporidium and Eimeria species prospering severity of disease outcome. Birds surviving to RV disease predominantly succumb to secondary bacterial infections, mostly E. coli and Salmonella spp. Recent developments in molecular tools including state-of-the-art diagnostics and vaccine development have led to advances in our understanding towards AvRVs. Development of new generation vaccines using immunogenic antigens of AvRV has to be explored and given due importance. Till now, no effective vaccines are available. Although specific as well as sensitive approaches are available to identify and characterize AvRVs, there is still need to have point-of-care detection assays to review disease burden, contemplate new directions for adopting vaccination and follow improvements in public health measures. This review discusses AvRVs, their epidemiology, pathology and pathogenesis, immunity, recent trends in diagnostics, vaccines, therapeutics as well as appropriate prevention and control strategies.

  2. Virus infections among young children--the first year of the INDIS study.

    Science.gov (United States)

    Simonen-Tikka, Marja-Leena; Klemola, Päivi; Suomenrinne, Sointu; Kaijalainen, Svetlana; Söderström, Daniela; Savolainen-Kopra, Carita; Näntö-Salonen, Kirsti; Ilonen, Jorma; Simell, Tuula; Simell, Olli; Roivainen, Merja

    2013-09-01

    The frequencies of early childhood infections were studied in healthy children with increased genetic risk for type 1 diabetes participating in the ongoing prospective high intensive infection follow-up Study, INDIS, started in 2009 in Turku, Finland. Here the results obtained from 160 stool to 160 nasal swab specimens collected in parallel at times of infectious symptoms in 2009-2010 from 45 children at the age of 24 months or younger are reported. The specimens were analyzed for enteric (human enterovirus, norovirus, rotavirus, sapovirus, astrovirus) and respiratory RNA viruses (human enterovirus and rhinovirus) common in early childhood, respectively, using highly validated virus-specific real-time PCR methods. According to the results 96% of the children had at least one virus infection during the study period and one or several viral agents were detected in 76% of sample sets. The most prevalent viral agents were human rhinovirus, enterovirus, parechovirus, and norovirus (genotype GII) with positive specimens 57.5%, 28.8%, 19.4%, and 6.9%, respectively. Other intestinal viruses were found in less than 2% of stool specimens. Single infections covered 40.0% of the specimens while multiple infections with two or more infectious agents were detected in 36.3% of specimens and altogether 11 combinations of viruses were included in the mixed infections. Although human enterovirus is known to be a frequent finding in stool specimens, especially during early childhood, it was found in this study more frequently in nasal swab specimens. Whether this is true, more general, in countries with the high hygiene level remains to be shown.

  3. Assessment of gastroenteric viruses from wastewater directly discharged into Uruguay River, Uruguay.

    Science.gov (United States)

    Victoria, M; Tort, L F L; García, M; Lizasoain, A; Maya, L; Leite, J P G; Miagostovich, M P; Cristina, J; Colina, R

    2014-06-01

    The aim of this study was to assess the viral contamination of group A rotavirus (RVA), norovirus (NoV), and human astrovirus (HAstV) in sewage directly discharged into Uruguay River and to characterize RVA genotypes circulating in Uruguay. For this purpose, sewage samples (n = 96) were collected biweekly from March 2011 to February 2012 in four Uruguayan cities: Bella Unión, Salto, Paysandú, and Fray Bentos. Each sample was concentrated by ultracentrifugation method. Qualitative and quantitative RT-PCR for RVA, NoV, and HAstV were performed. A wide dissemination of gastroenteric viruses was observed in the sewage samples analyzed with 80% of positivity, being NoV (51%) the most frequently detected followed by RVA with a frequency of 49% and HAstV with 45%. Genotypes of RVA were typed using multiplex semi-nested RT-PCR as follows: P[8] (n = 15), P[4] (n = 8), P[10] (n = 1), P[11] (n = 1), G2 (n = 29), and G3 (n = 2). The viral load ranged from 10(3) to 10(7) genomic copies/liter, and they were detected roughly with the same frequency in all participant cities. A peak of RVA and HAstV detection was observed in colder months (June to September), whereas no seasonality was observed for NoV. This study demonstrates for the first time, the high degree of gastroenteric viral contamination in the country; highlighting the importance of developing these analyses as a tool to determine the viral contamination in this hydrographic boundary region used by the local populations for recreation and consumption, establishing an elevated risk of gastroenteric diseases for human health.

  4. Cosavirus, Salivirus and Bufavirus in Diarrheal Tunisian Infants.

    Science.gov (United States)

    Ayouni, Siwar; Estienney, Marie; Hammami, Sabeur; Neji Guediche, Mohamed; Pothier, Pierre; Aouni, Mahjoub; Belliot, Gael; de Rougemont, Alexis

    2016-01-01

    Three newly discovered viruses have been recently described in diarrheal patients: Cosavirus (CosV) and Salivirus (SalV), two picornaviruses, and Bufavirus (BuV), a parvovirus. The detection rate and the role of these viruses remain to be established in acute gastroenteritis (AGE) in diarrheal Tunisian infants. From October 2010 through March 2012, stool samples were collected from 203 children CosV, SalV and BuV as well as for norovirus (NoV) and group A rotavirus (RVA) by molecular biology. Positive samples for the three screened viruses were also tested for astrovirus, sapovirus, adenovirus, and Aichi virus, then genotyped when technically feasible. During the study period, 11 (5.4%) samples were positive for one of the three investigated viruses: 2 (1.0%) CosV-A10, 7 (3.5%) SalV-A1 and 2 (1.0%) BuV-1, whereas 71 (35.0%) children were infected with NoV and 50 (24.6%) with RVA. No mixed infections involving the three viruses were found, but multiple infections with up to 4 classic enteric viruses were found in all cases. Although these viruses are suspected to be responsible for AGE in children, our data showed that this association was uncertain since all infected children also presented infections with several enteric viruses, suggesting here potential water-borne transmission. Therefore, further studies with large cohorts of healthy and diarrheal children will be needed to evaluate their clinical role in AGE.

  5. How long do nosocomial pathogens persist on inanimate surfaces? A systematic review

    Directory of Open Access Journals (Sweden)

    Schwebke Ingeborg

    2006-08-01

    Full Text Available Abstract Background Inanimate surfaces have often been described as the source for outbreaks of nosocomial infections. The aim of this review is to summarize data on the persistence of different nosocomial pathogens on inanimate surfaces. Methods The literature was systematically reviewed in MedLine without language restrictions. In addition, cited articles in a report were assessed and standard textbooks on the topic were reviewed. All reports with experimental evidence on the duration of persistence of a nosocomial pathogen on any type of surface were included. Results Most gram-positive bacteria, such as Enterococcus spp. (including VRE, Staphylococcus aureus (including MRSA, or Streptococcus pyogenes, survive for months on dry surfaces. Many gram-negative species, such as Acinetobacter spp., Escherichia coli, Klebsiella spp., Pseudomonas aeruginosa, Serratia marcescens, or Shigella spp., can also survive for months. A few others, such as Bordetella pertussis, Haemophilus influenzae, Proteus vulgaris, or Vibrio cholerae, however, persist only for days. Mycobacteria, including Mycobacterium tuberculosis, and spore-forming bacteria, including Clostridium difficile, can also survive for months on surfaces. Candida albicans as the most important nosocomial fungal pathogen can survive up to 4 months on surfaces. Persistence of other yeasts, such as Torulopsis glabrata, was described to be similar (5 months or shorter (Candida parapsilosis, 14 days. Most viruses from the respiratory tract, such as corona, coxsackie, influenza, SARS or rhino virus, can persist on surfaces for a few days. Viruses from the gastrointestinal tract, such as astrovirus, HAV, polio- or rota virus, persist for approximately 2 months. Blood-borne viruses, such as HBV or HIV, can persist for more than one week. Herpes viruses, such as CMV or HSV type 1 and 2, have been shown to persist from only a few hours up to 7 days. Conclusion The most common nosocomial pathogens may

  6. The structure of a rigorously conserved RNA element within the SARS virus genome.

    Directory of Open Access Journals (Sweden)

    Michael P Robertson

    2005-01-01

    Full Text Available We have solved the three-dimensional crystal structure of the stem-loop II motif (s2m RNA element of the SARS virus genome to 2.7-A resolution. SARS and related coronaviruses and astroviruses all possess a motif at the 3' end of their RNA genomes, called the s2m, whose pathogenic importance is inferred from its rigorous sequence conservation in an otherwise rapidly mutable RNA genome. We find that this extreme conservation is clearly explained by the requirement to form a highly structured RNA whose unique tertiary structure includes a sharp 90 degrees kink of the helix axis and several novel longer-range tertiary interactions. The tertiary base interactions create a tunnel that runs perpendicular to the main helical axis whose interior is negatively charged and binds two magnesium ions. These unusual features likely form interaction surfaces with conserved host cell components or other reactive sites required for virus function. Based on its conservation in viral pathogen genomes and its absence in the human genome, we suggest that these unusual structural features in the s2m RNA element are attractive targets for the design of anti-viral therapeutic agents. Structural genomics has sought to deduce protein function based on three-dimensional homology. Here we have extended this approach to RNA by proposing potential functions for a rigorously conserved set of RNA tertiary structural interactions that occur within the SARS RNA genome itself. Based on tertiary structural comparisons, we propose the s2m RNA binds one or more proteins possessing an oligomer-binding-like fold, and we suggest a possible mechanism for SARS viral RNA hijacking of host protein synthesis, both based upon observed s2m RNA macromolecular mimicry of a relevant ribosomal RNA fold.

  7. ENTEROPATHOGENS DETECTED IN A DAYCARE CENTER, SOUTHEASTERN BRAZIL: BACTERIA, VIRUS, AND PARASITE RESEARCH

    Directory of Open Access Journals (Sweden)

    Edna Donizetti Rossi Castro

    2015-02-01

    Full Text Available Introduction: The objective of this study was to determine the prevalence and etiological profile of enteropathogens in children from a daycare center. Methods: From October 2010 to February 2011 stool samples from 100 children enrolled in a government daycare center in the municipality of São José do Rio Preto, in the state of São Paulo, were collected and analyzed. Results: A total of 246 bacteria were isolated in 99% of the fecal samples; 129 were in the diarrheal group and 117 in the non-diarrheal group. Seventy-three strains of Escherichia coli were isolated, 19 of Enterobacter, one of Alcaligenes and one of Proteus. There were 14 cases of mixed colonization with Enterobacter and E. coli. Norovirus and Astrovirus were detected in children with clinical signs suggestive of diarrhea. These viruses were detected exclusively among children residing in urban areas. All fecal samples were negative for the presence of the rotavirus species A and C. The presence of Giardia lamblia, Entamoeba coli, Endolimax nana and hookworm was observed. A significant association was found between food consumption outside home and daycare center and the presence of intestinal parasites. Conclusions: For children of this daycare center, intestinal infection due to pathogens does not seem to have contributed to the occurrence of diarrhea or other intestinal symptoms. The observed differences may be due to the wide diversity of geographical, social and economic characteristics and the climate of Brazil, all of which have been reported as critical factors in the modulation of the frequency of different enteropathogens.

  8. Molecular epidemiological study on children under five years old with diarrhea and the long-term countermeasures for prevention and control of childhood diarrhea in Tangshan city%唐山市5岁以下儿童腹泻分子流行病学研究及长期预防控制对策

    Institute of Scientific and Technical Information of China (English)

    余亮科; 王林; 陈玉强; 高捷; 赵文喆

    2012-01-01

    Objective:To conduct molecular epidemiological study for children under five years old with diarrhea in Tangshan city, provide data for prevention and treatment of diarrhea, further put foreward long-term countermeasures for prevention and control of childhood diarrhea. Methods:The stool samples of 300 children with acute diarrhea were selected from Tangshan city randomly, after excluding 180 children with bacterial diarrhea, the other 120 children with viral diarrhea underwent rotavirus detection by colloidal gold method, RT-PCR was used to detect astrovirus, diarrheal adenovirus, and human calicivirus. Results; Among the children under five years old with viral diarrhea in Tangshan city, 16. 33% of them were infected by rotavirus, 10.33% of them were infected by astrovirus, 8. 67% of them were infected by diarrheal adenovirus, and 4. 67% of them were infected by human calicivirus. There was no statistically significant difference in the incidence of bacterial diarrhea among children in different age groups. The peak periods of childhood diarrhea induced by different viruses varied \\ the peak periods of childhood diarrhea induced by rotavirus, astrovirus, diarrheal adenovirus, and human calicivirus were 6-12 months, 12-28 months, 12-24 months, and 12-18 months, respectively. The change of season had no obvious impact on the causes of diarrhea. Conclusion:Bacterial infection was the most important cause of childhood diarrhea, but for the children with viral diarrhea, rotavirus was the main induced cause. More attention should be paid to the children during every age period to prevent and control bacterial diarrhea, childhood diarrhea induced by different viruses can be treated with corresponding measures according to different peak periods of onset age.%目的:对唐山市5岁以下儿重腹泻进行分子流行病学研究,为腹泻的防治提供数据,进而提出对儿重腹泻的长期预防控制措施.方法:随机抽取唐山市300例儿童急性腹泻的

  9. Norovirus diversity in diarrheic children from an African-descendant settlement in Belem, Northern Brazil.

    Directory of Open Access Journals (Sweden)

    Glicélia Cruz Aragão

    Full Text Available Norovirus (NoV, sapovirus (SaV and human astrovirus (HAstV are viral pathogens that are associated with outbreaks and sporadic cases of gastroenteritis. However, little is known about the occurrence of these pathogens in relatively isolated communities, such as the remnants of African-descendant villages ("Quilombola". The objective of this study was the frequency determination of these viruses in children under 10 years, with and without gastroenteritis, from a "Quilombola" Community, Northern Brazil. A total of 159 stool samples were obtained from April/2008 to July/2010 and tested by an enzyme immunoassay (EIA and reverse transcription-polymerase chain reaction (RT-PCR to detect NoV, SaV and HAstV, and further molecular characterization was performed. These viruses were detected only in the diarrheic group. NoV was the most frequent viral agent detected (19.7%-16/81, followed by SaV (2.5%-2/81 and HAstV (1.2%-1/81. Of the 16 NoV-positive samples, 14 were sequenced with primers targeting the B region of the polymerase (ORF1 and the D region of the capsid (ORF2. The results showed a broad genetic diversity of NoV, with 12 strains being classified as GII-4 (5-41.7%, GII-6 (3-25%, GII-7 (2-16.7%, GII-17 (1-8.3% and GI-2 (1-8.3%, as based on the polymerase region; 12 samples were classified, based on the capsid region, as GII-4 (6-50%, being 3-2006b variant and 3-2010 variant, GII-6 (3-25%, GII-17 (2-16.7% and GII-20 (1-8.3%. One NoV-strain showed dual genotype specificity, based on the polymerase and capsid region (GII-7/GII-20. This study provides, for the first time, epidemiological and molecular information on the circulation of NoV, SaV and HAstV in African-descendant communities in Northern Brazil and identifies NoV genotypes that were different from those detected previously in studies conducted in the urban area of Belém. It remains to be determined why a broader NoV diversity was observed in such a semi-isolated community.

  10. Human bocavirus in hospitalized children with acute gastroenteritis in Russia from 2010 to 2012.

    Science.gov (United States)

    Tymentsev, Alexander; Tikunov, Artem; Zhirakovskaia, Elena; Kurilschikov, Alexander; Babkin, Igor; Klemesheva, Vera; Netesov, Sergei; Tikunova, Nina

    2016-01-01

    Human bocavirus (HBoV) can cause respiratory diseases and is detectable in the stool samples of patients with gastroenteritis. To assess the prevalence of HBoV in children hospitalized with acute gastroenteritis in Novosibirsk, Russia, as well as its genetic diversity and the potential role in the etiology of gastroenteritis in this region, a total of 5502 stool samples from children hospitalized with gastroenteritis from 2010 to 2012, n=5250, and healthy children, n=252, were assayed for the presence of HBoV DNA by semi-nested PCR. The HBoV DNA was found in 1.2% of stool samples from children, with gastroenteritis varying from 0.5% in 2012 to 1.7% in 2011. The prevalence of HBoV in healthy children was 0.3%. HBoV strains were detected throughout the year with an increase in the fall-winter season. In 87% of cases, HBoV was detected in children before 1 year of age. All known HBoV genetic variants have been detected in Novosibirsk, although with different prevalences: HBoV2>HBoV1>HBoV4>HBoV3. At the beginning of 2011, HBoV2 replaced HBoV1 as the most prevalent variant. The median age of children with detected HBoV1 was 8.3months, and that with HBoV2 was 8.0 months. All HBoV-positive samples were assayed for the presence of the rotaviruses A and C, norovirus GII, astrovirus, enterovirus, adenovirus F, Salmonella spp., Campylobacter spp., Shigella spp., and EIEC. HBoV1 and HBoV2 as single agents were found in 45.8% and 60% samples, respectively, although this difference was not statistically significant. In the case of co-infections, HBoV was most frequently recorded with rotavirus A and norovirus GII. This study demonstrated that the detection rate of HBoV in stool samples from children with gastroenteritis was low, although both HBoV1 and HBoV2 could be found as the sole agents in children with gastroenteritis in Novosibirsk.

  11. [Rotavirus and other viruses of diarrhea].

    Science.gov (United States)

    Bajolet, O; Chippaux-Hyppolite, C

    1998-01-01

    Rotaviruses represent 80% of recognized viral etiologies and 140 million cases of diarrhea per year. They strike young children with similar frequency throughout the world, but the mortality rate is high in developing countries only, with some 870,000 deaths per year (WHO, 1997). Rotaviruses belong to the family of Reoviridae; they are segmented bicatenary RNA viruses, which explains their genetic variability, the presence of mixed infections, the establishment for some time already of a molecular epidemiology by electrophore types. The viruses are "naked" and thus resistant to the outside environment; their massive elimination, 10(8) to 10(10)viral particles per gram of faeces, begins with the first day of diarrhea. They are found in used water and can also be concentrated by shellfish; the environment thus constitutes a notable reservoir for the virus. Oral-faecal transmission is facilitated by deficient sanitary conditions. The 11 fragments of the genome each codify for 1 viral protein; 2 surface proteins, VP4 and VP7, bring about the formation of neutralizing antibodies, which are important for the protection and determination of different serotypes. A non structural protein--NSP4--would seem to intervene in the cytopathogenic effect and may act as a veritable viral enterotoxine. Numerous animal species are infected by rotaviruses which are district from the human ones. The pathology as it affects animals is of economic importance and of interest as an experimental and vaccinal model. Between human and animal rotaviruses there can be genetic rematchings and the VP6 protein is an antigen common to the group. The description of the other viruses responsible for diarrhea has benefited from widespread use of electronic microscopes from the very first years of study of rotaviruses. These other viruses belong to 6 different types: adenovirus, calcivirus, astrovirus, Norwalk agent and related viruses, coronavirus, enterovirus. They therefore have a structural and

  12. Comportamiento de la diarrea causada por virus y bacterias en regiones cercanas a la zona ecuatorial

    Directory of Open Access Journals (Sweden)

    María Fernanda Gutiérrez

    2005-12-01

    Full Text Available Introducción: La enfermedad diarreica aguda (EDA es un problema de salud a nivel mundial que afecta a la población infantil de distintas regiones. Casi todos los estudios epidemiológicos se han hecho en países con estaciones y poco se informa su comportamiento en países sin estaciones, donde la EDA es endémica con picos epidémicos. Objetivos: Contribuir a conocer la conducta de EDA en Colombia y determinar si su comportamiento es diferente en niños menores de cinco años en dos regiones distintas entre sí en geografía y clima. Materiales y métodos: Se hizo un estudio descriptivo en dos localidades colombianas. Una en la costa atlántica y otra en el centro del país. La muestra se obtuvo en menores de cinco años que consultaron por diarrea a centros asistenciales de cada región. Los microorganismos bacterianos se identificaron mediante pruebas bioquímicas y los virus con técnicas inmunoenzimáticas. En el análisis estadístico se siguieron un ensayo bivariado y pruebas Z de normalidad para verificar si el clima modifica el comportamiento de EDA y si se presenta de manera distinta en las dos regiones. Resultados: En ambas zonas (Cartagena, Bolívar y Facatativa, Cundinamarca predominó la diarrea viral, frente a la EDA bacteriana. También en ambas el rotavirus fue prevalente. Fue mucho más baja la presencia de astrovirus y adenovirus. No hubo datos con significación estadística para demostrar que las condiciones ambientales y las propias de los niños, alteran el comportamiento de la EDA, pero sí se observó que la EDA por rotavirus se comporta de manera diferente al analizar en forma comparativa las dos regiones del estudio.

  13. Comportamiento de la diarrea causada por virus y bacterias en regiones cercanas a la zona ecuatorial.

    Directory of Open Access Journals (Sweden)

    María Fernanda Gutiérrez

    2009-11-01

    Full Text Available INTRODUCCIÓN: La enfermedad diarreica aguda (EDA es un problema de salud a nivel mundial que afecta a la población infantil de distintas regiones. Casi todos los estudios epidemiológicos se han hecho en países con estaciones y poco se informa su comportamiento en países sin estaciones, donde la EDA es endémica con picos epidémicos. OBJETIVOS: Contribuir a conocer la conducta de EDA en Colombia y determinar si su comportamiento es diferente en niños menores de cinco años en dos regiones distintas entre sí en geografía y clima. MATERIALES Y MÉTODOS: Se hizo un estudio descriptivo en dos localidades colombianas. Una en la costa atlántica y otra en el centro del país. La muestra se obtuvo en menores de cinco años que consultaron por diarrea a centros asistenciales de cada región. Los microorganismos bacterianos se identificaron mediante pruebas bioquímicas y los virus con técnicas inmunoenzimáticas. En el análisis estadístico se siguieron un ensayo bivariado y pruebas Z de normalidad para verificar si el clima modifica el comportamiento de EDA y si se presenta de manera distinta en las dos regiones. RESULTADOS: En ambas zonas (Cartagena, Bolívar y Facatativa, Cundinamarca predominó la diarrea viral, frente a la EDA bacteriana. También en ambas el rotavirus fue prevalente. Fue mucho más baja la presencia de astrovirus y adenovirus. No hubo datos con significación estadística para demostrar que las condiciones ambientales y las propias de los niños, alteran el comportamiento de la EDA, pero sí se observó que la EDA por rotavirus se comporta de manera diferente al analizar en forma comparativa las dos regiones del estudio.

  14. 2011年西宁地区儿童腹泻病毒流行病学调查%Epidemiological investigation on diarrhea virus among children in Xining area in 2011

    Institute of Scientific and Technical Information of China (English)

    刘桂香; 易虎; 田登; 赵生仓; 杨维成; 徐琼; 石燕; 张华一

    2013-01-01

    Objective: To conduct an epidemiological investigation on diarrhea virus in Xining city in 2011, analyze epidemic strains and trend of diarrhea virus in Qinghai province, and provide experimental data for diarrhea induced by diarrhea virus to control prevalence of diarrhea virus. Methods: Nest - PCR and multiplex PCR were used to detect diarrhea virus among 488 infants in Xining area. Results : The detection rates of diarrhea virus in children with diarrhea in Xining area; group A rotavirus (6. 76% , 33 children, including 15 children of G9 type, 13 children of G3 type, 1 child of P6 type, 2 children of P8 type, and 2 children of P9 type) , group B/C rotavirus (0. 61% , 3 children) , calicivirus (4. 71% , 23 children, including 17 children with sapovirus and 6 children with norovirus of GⅡ type) , astrovirus (1. 43% , 7 children) . Conclusion: Group A rotavirus and calicivirus are main pathogens of infantile diarrhea in Xining area.%目的:对2011年西宁市腹泻病毒进行流行病学调查,对青海省腹泻病毒流行株和流行趋势进行统计分析,为腹泻病毒引起的腹泻提供实验室资料,更好地控制病毒性腹泻的流行.方法:用巢式PCR和多重PCR方法对西宁地区488例婴幼儿腹泻标本进行腹泻病毒检测.结果:西宁地区腹泻儿童中腹泻病毒的检出率分别为:轮状病毒A组阳性33例(6.76%),其中G9型15例、G3型13例、P6型1例、P8型2例、P9型2例;轮状病毒B/C组阳性3例(0.61%);杯状病毒阳性23例(4.71%),其中扎如病毒17例、诺如病毒GⅡ型6例;星状病毒阳性7例(1.43%).结论:轮状病毒A组和杯状病毒是西宁地区婴幼儿腹泻的主要病原体之一.

  15. Viruses of foodborne origin: a review

    Directory of Open Access Journals (Sweden)

    Todd EC

    2015-04-01

    Full Text Available Ewen CD Todd,1,2 Judy D Greig3 1Ewen Todd Consulting LLC, Okemos, MI, USA; 2Department of Nutrition and Food Science, Faculty of Agricultural and Food Sciences, American University of Beirut, Beirut, Lebanon; 3Division of Public Health Risk Sciences, Laboratory for Foodborne Zoonoses, Public Health Agency of Canada, Guelph, ON, Canada Abstract: Enteric viruses are major contributors to foodborne disease, and include adenovirus, astrovirus, rotavirus, sapovirus, hepatitis A and E viruses, and norovirus. From a foodborne transmission perspective, norovirus is the most important; however, hepatitis A is associated with more serious illness. Foodborne viruses are transmitted through contaminated food, but also in combination with person-to-person contact or through environmental contamination. These viruses survive well in the environment, are excreted in abundance in feces, and have a low infectious dose, all of which facilitate spread within a community. Many colonized individuals experience mild gastroenteritis lasting a few days or are asymptomatic, although viral excretion may continue over days or weeks. Severe illness tends to be restricted to the very young and elderly, especially in closed communities such as schools and homes for the aged. In the USA, norovirus is considered to be responsible for two thirds of all foodborne illnesses occurring in a wide range of institutional settings, including schools, colleges, child care centers, cruise ships, prisons, and soldiers on campaign. Norovirus outbreaks also occur at one-time events, such as banquets, wedding receptions, birthday parties, and potluck meals, and are most often introduced by infected food workers producing, preparing, or serving food, or through self-service buffets. Often the infections are introduced from the community into institutions where they can infect the majority of residents unless quickly controlled. In countries where economic assessments have been completed

  16. VIRAL ETIOLOGY ACUTE INTESTINAL INFECTIONS MOLECULAR MONITORING IN CHILDREN’S HOSPITAL

    Directory of Open Access Journals (Sweden)

    A. V. Sergeeva

    2015-01-01

    infections in patients with clinical signs of intestinal infection, and contact persons was 43.8%. The etiological structure of intestinal virus infections was presented by noroviruses (73.2% genotypes GII.1, GII.3, GII.4 Sydney 2012, rotaviruses (23.2% genotypes G4P[8] and G1-IP[8], and adenovirus (1.8% of the group F and astroviruses (1.8% genotypes 1 and 2. Among the hospitalized children were 9 cases of viral etiology acute intestinal infections importation, and 66 of cases were of nosocomial origin. Examination of the environment revealed the presence of contamination by enteric viruses in 47.8% of cases. 

  17. Etiological analysis of diarrhea patients visiting enteric disease clinics in Beijing%北京市肠道门诊腹泻患者病原谱分析

    Institute of Scientific and Technical Information of China (English)

    刘晓峰; 张扬; 张兰荣; 甄博珺; 李朝辉; 吴阳博; 高志勇

    2015-01-01

    目的:了解北京市肠道门诊腹泻患者的病原学特征。方法2013年7月至2014年6月在2家哨点医院肠道门诊收集腹泻患者的粪便标本595份。对沙门菌、志贺菌、副溶血性弧菌、O1和O139群霍乱弧菌、致泻大肠埃希菌进行分离培养,对轮状病毒、诺如病毒、星状病毒和肠道腺病毒进行核酸检测。采用描述性流行病学方法对病原的时间分布、人群分布、血清型别等特征进行分析。组间率的比较采用χ2检验。结果595份样本中,128份细菌检测阳性,检出率为21.5%,其中致泻大肠埃希菌检出率为11.4%(68/595),其次为副溶血性弧菌(6.9%,41/595)、沙门菌(2.4%,14/595)和志贺菌(2.2%,13/595),霍乱弧菌未检出。112份样本腹泻病毒核酸检测阳性,检出率为18.8%,其中诺如病毒检出率为9.1%(54/595),其次为轮状病毒(8.7%,52/138),星状病毒(1.8%,11/595)和肠道腺病毒(0.7%,4/595)。致泻大肠埃希菌以肠致病性大肠埃希菌、肠产毒素性大肠埃希菌、肠黏附性大肠埃希菌为主,副溶血性弧菌血清型别主要为O4∶K8。6~9月份为病原菌检出的高峰期,当年11月至次年3月为腹泻病毒检出的高峰期。结论诺如病毒、轮状病毒亦是肠道门诊腹泻患者的主要病原,应予以重视。%Objective To investigate the etiological characteristics of diarrhea patients visiting enteric disease clinics in Beijing .Methods A total of 595 stool samples were collected among outpatients with diarrhea vistiting enteric disease clinics at two sentinel hospitals from July 2013 to June 2014 . Diarrheagenic Escherichia coli (E . coli) , Vibrio parahemolyticus , O1 or O139 Vibrio cholerae , Salmonella and Shigella were isolated according to standard methods . And rotavirus , norovirus , astrovirus and enteric adenovirus were identified by

  18. Molecular epidemiology of viral diarrhea in Chengdu infants and young children%成都地区婴幼儿病毒性腹泻分子流行病学研究

    Institute of Scientific and Technical Information of China (English)

    谢晓丽; 张伟; 廖雪春; 邓孝智; 刘丽荣; 张琪琛; 商丽红; 温燕; 任敏

    2012-01-01

    目的 通过分子流行病学研究成都地区婴幼儿病毒性腹泻的病原学特点,掌握本地区病原分布特征,为疫苗研制和疫情控制提供科学依据.方法 采用酶联免疫吸附试验( ELISA)及逆转录聚合酶链反应(RT-PCR)对成都地区2006 -2008年度376例婴幼儿腹泻粪便标本进行轮状病毒(RV)、杯状病毒(HuCV)、星状病毒(AstV)及肠道腺病毒(Adv)检测.结果 RV的检出率为37.76%(142/376),其中42例进行G分型,45例进行P分型,G分型以G3型为主21例(50%),其次为G2和G1型,P分型以P[8]型为主21例,其次是P[4]型19例.RV感染主要为6~23月的婴幼儿,发病高峰在10 ~12月份(75.8%).RT-PCR法检出HuCV、AstV及Adv的检出率分别为15.85%、1.64%及2.04%.结论 RV是成都地区婴幼儿病毒性腹泻的主要病原,其流行的主要血清型为G3型、P[8]、P[4]型.除RV外,HuCV也是重要的病原.%Objective To investigated the molecular epidemiologic features of viral diarrhea in Chengdu infants and young children,and to establish baseline patterns of etiology,provides the scientific basis for the vaccine development and the epidemic situation control.Methods From March,2006 to December,2008,a total of 376 infants and young children from Chengdu area hospitalized for diarrhea in Chengdu Children's Hospital were enrolled in this study.The stool specimen collected from each patient was tested for rotavirus ( RV),Calicivirus ( CV),astrovirus ( AstV ) and adenovirus (Adv) by using enzyme linked immunosorbent assay (ELISA) and reverse transcription-polymerase chain reaction (RT-PCR) examination.Results Among those 376 cases,there were 142 cases (37.76%) of RV infections,which scattered predominantly in October to December.Among 234 cases RV negativity,there were 29 cases HuCV infections ( 15.85% ),5 cases AstV infections ( 1.64% ),and 8 cases Adv infections (2.04%).Conclusion RV appeared to be the main etiological agent of viral diarrhea in Chengdu

  19. Etiological study of enteric viruses and bacteria in adult diarrhea in Chongqing area%重庆地区成人感染性腹泻病原学分析

    Institute of Scientific and Technical Information of China (English)

    娄茜; 孙滨; 张再宽; 张秀瑜; 王云英; 黄长武; 陈维贤

    2012-01-01

    目的 研究重庆地区成人感染性腹泻患者的病原学特点.方法 采用MICROSCAN系统进行粪便细菌培养,用ELISA和多重PCR方法进行病毒检测.结果 130例腹泻标本中,检出细菌阳性17例,包括沙门菌属7例,志贺菌属5例,副溶血弧菌3例和嗜水气单胞菌2例.病毒检测中,单重感染30例,双重感染10例,三重感染1例.A组轮状病毒检出率为3.85% (5/130),B轮状病毒检出率为3.85% (5/130),C组轮状病毒检出率为15.4% (20/130),诺如病毒检出率为9.23% (12/130),星状病毒检出率为4.62%(6/130),札如病毒检出率为3.08% (4/130),腺病毒检出率为0.77%(1/130).结论 重庆地区成人腹泻患者中,沙门菌和志贺菌为细菌感染的主要菌种,轮状病毒和诺如病毒为病毒感染的主要病原体.%Objective To study the etiologic characteristics of enteric viruses and bacteria in adult diarrhea in Chongqing area. Methods Bacteria were isolated from feces specimens and indentified by MICROSCAN system. Viruses were detected with ELISA and multiple polymerase chain reactions ( PCR). Results A total of 17 strains of pathogenic bacteria were isolated from 130 specimens, including 7 strains of Salmonella, 5 strains of Shigella, 3 strains of vice Hemolysis vibrio and 2 strains of Aeromonas hydrophilia. Among the virus detection results, the positive rates for Rotavirus A, Rotavirus B, Rotavirus C, Norovirus, Astrovirus, Sapporo like virus and Adenovirus were 3. 85% (5/130) , 3. 85% (5/130) , 15.4% (20/130) , 9.23% (12/130), 4.62% (6/130), 3.08% (4/130) and 0. 77% (1/130), respectively. Conclusion The prevalent pathogens for adult diarrhea in Chongqing area are Salmonella, Shigella, Rotavirus and Norovirus.

  20. 人星状病毒非结构蛋白酵母双杂交诱饵表达载体构建及酵母转化子特性鉴定%Construction of Yeast Two-Hybrid Bait Vector of HAstV Non-Struc-tural Protein and Characterization of Yeast Transformants

    Institute of Scientific and Technical Information of China (English)

    李攀; 崔成成; 杨思达; 黄芬; 曾韦锟; 井申荣

    2014-01-01

    Objective To construct a yeast two-hybrid bait vector of non-structure protein 1a/3 (nsP1a/3) of human astrovirus (HAstV), and to characterize the yeast transformants after elec-trotransformation of the recombinant plasmid into the yeast competent cells .Methods The coding sequence of HAstV nsP 1 a/3 obtained by PCR was inserted into the modified plasmid pGST 7.The re-combinant vector nsP1a/3-pGBST7 confirmed by DNA sequencing was transformed into the Y 2 HGo-ld yeast competent cells .The cytotoxicity of the expression products of the recombiant plasmid nsP1a/3-pGBST7 and the autoactivation of the report gene in yeast transformants were exam-ined.Results The sequencing of the nsP 1 a/3-pGBST7 vector showed that the recombinant bait plasmid was successfully constructed and the open reading frame of nsP 1a/3 insert was correct.After electrotransformation, it was found that the expression products of the nsP 1a/3-pGBST7 vector had no cytotoxicity to yeast transformants and also they did not activate the report gene .Conclusion The yeast two-hybrid bait vector of HAstV nsP 1 a/3 was successfully constructed , which laid a foun-dation for further studying the interaction of HAstV nsP 1 a/3 with target proteins and the replication mechanism of HAstV in host cells .%目的:构建人星状病毒非结构蛋白3( non-structure protein 1a/3, nsP1a/3)基因酵母双杂交诱饵重组质粒,电转化酵母感受态细胞后鉴定酵母转化子特性。方法聚合酶链反应扩增人星状病毒nsP1a/3编码的基因序列,定向插入到穿梭表达载体pGBKT7改造后的pGBST7中,测序确定无误后将其电转化到Y2HGold酵母感受态细胞中,对细胞特性即nsP1a/3-pGBST7诱饵质粒表达产物的毒性和报告基因自激活作用进行鉴定。结果测序结果表明诱饵重组质粒构建正确,阅读框无误。电转化酵母后,其表达产物对酵母细胞无毒性作用,对报告基因亦无激活作用。结论成功构建了

  1. Surveillance and analysis on diarrheal disease pathogen spectrum among children under 5 years old in Pudong New Area of Shanghai City%上海市浦东新区5岁以下儿童腹泻病病原谱监测分析

    Institute of Scientific and Technical Information of China (English)

    费怡; 孙乔; 傅益飞; 朱渭萍; 薛曹怡

    2013-01-01

    [目的]了解上海市浦东新区5岁以下儿童腹泻病的病原谱构成、病原特征,为制定预防控制措施提供依据。[方法]对2012年2月-2013年1月浦东新区人民医院、儿童医学中心2家监测点医院的619例腹泻患儿进行个案调查,同时采集粪便标本进行肠道致病菌和病毒的检测。[结果]诺如病毒、轮状病毒、肠道腺病毒、扎如病毒、星状病毒的检出率分别为11.95%、7.27%、2.91%、1.45%、0.97%;致泻性大肠埃希菌、非伤寒沙门菌、弯曲菌、嗜水气单胞菌、志贺菌、小肠结肠炎耶尔森菌的检出率分别为5.65%、2.42%、0.81%、0.65%、0.48%、0.32%。病毒检出率8月至次年的3月为27.70%,高于其他月份的18.48%(χ2=7.18,P<0.05);细菌检出率4-10月为15.34%,远高于其他月份的3.15%(χ2=31.22,P<0.05)。[结论]诺如病毒为浦东新区5岁以下儿童病毒性腹泻的最主要病原,致泻性大肠埃希菌为细菌性腹泻的最主要病原。细菌性腹泻的防控应以夏秋季为重点,病毒性腹泻应以全年为重点。%[Objective] To investigate the pathogen spectrum and characteristics of diarrheal disea-ses among children under five and to provide the basis for prevention and control measures . [ Methods] From February 2012 to January 2013, a total of 619 cases from 2 sentinel hospitals were collected for case survey .From them were collected stool specimens for detection of enteric pathogenic bacteria and virus nu-cleic acids. [Results] The virus detection rate was norovirus (11.95%), rotavirus(7.27%), enteric adenovirus(2.91%), saporovirus(1.45%) and astrovirus(0.97%).The bacteria detection rate was E. coli(5.65%), nontyphoidal salmonella (2.42%), campylobacter (0.81%), aeromonas hydrophila (0.65%),shigella(0.48%) and yersinia enterocolitica (0.32%).The virus detection rate was 27.70%from August to next March

  2. 2014年无锡市感染性腹泻的病原学分析%Etiological analysis of infectious diarrhea in Wuxi,2014

    Institute of Scientific and Technical Information of China (English)

    管红霞; 肖勇; 沙丹

    2016-01-01

    Objective To understand the pathogen spectrum and epidemiological characteristics of infectious diarrhea in Wuxi. Methods The stool specimens from outpatients with diarrhea were collected from sentinel hospitals from January to December 2014. The nucleic acid of norovirus,rotavirus,enteric adenovirus,human astrovirus and sapovirus was determined by real-time fluorescence polymerase chain reaction(PCR). Diarrheagenic Escherichia coli,Salmonella spp,Shigella spp,Vibrio parahaemolyticus and Vibrio cholerae were identified by conventional culture methods. Pathogenic composition,seasonal variation and population distribution were analyzed. Results A total of 915 specimens were monitored. The positive rate of viral nucleic acid was 14.4%,and norovirus(9.4%) and rotavirus(3.4%) showed obvious seasonal distribution characteristics with the peak incidence occurring in winter and spring. The infection rate of children of 0-5 years old was higher than those of the other age groups. Bacteria,with the positive rate 15.0%,were isolated from June to September. Salmonella spp(6.0%) was the main isolate whose predominant serotypes were Salmonella enteritidis and Salmonella typhimurium,and the second was diarrheagenic Escherichia coli(4.6%). Conclusions The viral diarrhea deserves attention in winter and spring,norovirus and rotavirus are the common pathogens. The bacterial diarrhea is given priority in summer,whose dominant pathogens are Salmonella spp and diarrheagenic Escherichia coli. Children under 5 years old are main for the prevention and control of viral diarrhea.%目的:了解无锡市感染性腹泻的病原谱及流行特征。方法连续采集2014年1至12月无锡市腹泻监测哨点医院门诊腹泻病例的粪便样本,采用实时荧光聚合酶链反应(PCR)检测诺如病毒、轮状病毒、肠道腺病毒、人星状病毒和札如病毒的核酸,采用传统的分离培养方法检测致泻性大肠埃希菌、沙门菌、志贺菌、副溶血性

  3. Establishment and Application of a RT-PCR Assay for Detection of Newly Emerged Porcine Deltacoronavirus%新现猪Delta冠状病毒RT-PCR检测方法的建立及其应用

    Institute of Scientific and Technical Information of China (English)

    张帆帆; 唐玉新; 宋德平; 周信荣; 黄冬艳; 李安琪; 彭棋; 陈燕君; 吴琼; 何后军

    2016-01-01

    the result of multialignment analysis on the published PDCoV genome sequences on GenBank database, a pair of primers specific to the conserved nucleocapsid (N) gene of PDCoV was designed by using the online software Primer 3.0, and then a RT-PCR for detection of PDCoV was established by utilizing the designed primers. Diarrheal samples were tested by the established RT-PCR assay, and several positive PCR products were selected for cloning and sequencing. To analyze the phylogenetic relationships between PDCoV and porcine epidemic diarrhea virus (PEDV) as well as transmissible gastroenteritis virus (TGEV), a phylogenetic tree was constructed based on the partial N gene sequences of PDCoV obtained in this study and 18 reference PDCoV sequences from other countries/areas, and corresponding N gene sequences of representative PEDV and TGEV strains. To determine the specificity of the RT-PCR assay established, RNAs of PEDV, TGEV, porcine kobuvirus (PKoV), porcine astrovirus (PAstV), porcine reproductive and respiratory syndrome virus (PRRSV) and classical swine fever virus (CSFV) were tested. To evaluate the detection limit of the assay, a recombinant plasmid containing the N gene fragment was constructed and served as templates with 10-fold serial dilution starting from 1.0×106 copies/μL to 1.0×101 copies/μL. Clinic diarrheal samples of pigs (n=249) from sows and piglets collected in Jiangxi Province from 2012 to 2015 were tested for the presence of PDCoV by the established RT-PCR, and some PDCoV positive amplicons were cloned and then sequenced for the specificity confirmation of the assay.[Result]A single expected DNA fragment band of 329 bp in size was amplified by using the designed primers. Sequences of the amplicons selected were subjected to a BLAST search against the GenBank database, and the results indicated that the sequences hit PDCoV with a nucleotide identity as high as 99.1%, demonstrating that the amplified targets were PDCoV. A phylogenetic tree was