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PubMed2005

Etiologic agents of acute gastroenteritis among Japanese infants and children: virus diversity and genetic analysis of sapovirus.

A total of 371 fecal specimens from infants and children with acute gastroenteritis in Maizuru, Tokyo and Osaka, Japan from July 2002 to June 2003 were tested for the presence of diarrheal viruses by reverse transcription-polymerase chain reaction (RT-PCR), reverse passive hemagglutination (PRHA), RNA-polyacrylamide gel electrophoresis (PAGE), latex agglutination and sequence analysis methods. Among diarrheal viruses detected, group A rotavirus was the most prevalent (42.2%) followed by norovirus (28.9%), group C rotavirus (8.4%), sapovirus (6.7%), adenovirus (5.3%) and astrovirus (0.9%), respectively. There was the high rate (7.6%) of viral mixed infections. Sapovirus was classified into 6 genotypes (GI/1, GI/4, GI/5, GI/6 and GII/1 and one novel tentatively called GII/5). It is noteworthy that genogroup II sapovirus can be classified into 5 genotypes. Our findings confirmed the presence of many diarrheal viruses co-circulating among Japanese infants and children and showed the great genetic diversity among sapoviruses.

Acute Disease↗
PubMed1990

[Atypical rotavirus isolated from acute infectious nonbacterial gastroenteritis in Fukuoka].

An outbreak of epidemic diarrhea (nonbacterial acute gastroenteritis) in Fukuoka was studied electron microscopically and electrophoretically. From the result of electron microscopy, rotavirus particles about 70 nm in diameter were observed in fecal samples. However, the serological study did not reveal any antigenic relatedness to the ordinary rotaviruses by RPHA. The outbreak appeared to be caused by group C rotaviruses, based on the pattern of the RNA genome segments in polyacrylamide gels.

Acute Disease↗
PubMedDate not supplied

[Rotavirus etiology of hospital cases of acute gastroenteritis].

The etiology of enteric illnesses occurring during hospitalization in children admitted in January-May, 1985, for respiratory tract illnesses was studied by several methods including electron microscopy (EM), enzyme immunoassay (EIA), and PAG electrophoresis. Rotaviruses were detected in 22 (44.9%) t of 49 children with symptoms of intestinal infection, in February in 11 (84.6%) out of 13 patients. The analysis of virus genome RNA of 7 isolates positive in EM and EIA revealed in all the isolates an identical pattern of RNA segment distribution typical of "long" variants of group A rotaviruses. The appurtenance of the isolates to the same electrophoretic type together with epidemiological data allows the examined cases of rotavirus gastroenteritis to be considered as nosocomial ones. A sporadic case of gastroenteritis is described induced by a rotavirus antigenically different from Group A rotaviruses and having the electrophoretic type similar to that described for group C rotaviruses.

Acute Disease↗
PubMedDate not supplied

[Rotavirus, adenovirus, astrovirus, calicivirus and small round virus particles in feces of children with and without acute diarrhea, from 1987 to 1988, in the greater São Paulo].

Between 1987 and 1988, 193 faecal specimens from children, with or without diarrhea, were submitted to enzyme immunoassay, polyacrylamide-gel electrophoresis and electronmicroscopy tests for virus detection. The positivity for Rotavirus, Adenovirus, Astrovirus, Calicivirus and Small Round Virus Particles (SRVP) was 11.3%, 3.1%, 2.1%, 1.0% and 4.1%, respectively, for the 97 children with acute diarrhea. Of the 96 children without diarrhea, 4.2% were positive for Rotavirus, 1.0% for Calicivirus and 7.3% for SRVP. Of 15 positive specimens for Rotavirus, 14 showed electrophoretic patterns proper to group A and 1 specimen of group C Rotavirus. The analysis of electrophorotypes demonstrated great heterogeneity of electrophoretic patterns and predominance of subgroup 2, "long". The association of virus, bacteria and parasites was present both in children with or without acute diarrhea.

Acute Disease↗
PubMed1993

Sequence analysis of three non structural proteins of a porcine group C (Cowden strain) rotavirus.

Sequences of three gene products of a group C (Cowden strain) rotavirus are presented and compared with the sequences of the corresponding group A (SA11) proteins. The degree of similarity for gene 7, 9, and 10 is respectively 34%, 58%, and 45%. Comparison of these 2 viruses allowed to identify several regions well conserved. In the protein coded by Cowden segment 7 (NS 53) only a short cystein and histidine rich region, presenting the zinc finger consensus motif, is common to group A (segment 5) and group C sequences. Conversely the protein coded by segment 9 (NS 35) presented marked homology with group A NS 35. The protein coded by segment 10 (NS 26) is serine rich and presents an accumulation of charged residues near the carboxy terminus, like group A counterpart. This genomic segment presented a single large open reading frame, that contrasts with the group A counterpart for which a second out of phase ORF is used in rotavirus infected MA 104 cells.

Amino Acid Sequence↗
PubMed1983

Survival of enteric viruses adsorbed on electropositive filters.

Three viruses (poliovirus type 1, rotavirus SA-11, and bacteriophage f2) adsorbed on electropositive microporous filters survived at least 5 weeks at 4 degrees C. Poliovirus type 1 and bacteriophage f2 also survived at least 6 weeks at -20 degrees C. Rotavirus SA-11 was not recovered after 1 week at -20 degrees C. The stability of viruses adsorbed on electropositive filters may enable extensive monitoring of viruses in water.

Adsorption↗
PubMed1987

Physicochemical characterization of porcine pararotavirus and detection of virus and viral antibodies using cell culture immunofluorescence.

A cell culture immunofluorescence (CCIF) assay was optimized for detection of porcine pararotavirus (group C rotavirus) in intestinal contents. The greatest viral infectivity was observed when MA104 cells (5 days after subculturing) were rinsed and refed in serum-free medium before inoculation, pancreatin was added to the inocula, and the inocula were centrifuged onto the cells. Gentamicin treatment of pararotavirus samples to reduce bacterial contamination also reduced the viral infectivity of these samples for MA104 cells. An indirect CCIF assay was used to determine the prevalence of pararotavirus and rotavirus antibodies in pig sera. In pigs from four herds, pararotavirus antibodies were detected in 100% (68 of 68) of adults and 59% (24 of 41) of weanling pigs, while 86% (24 of 28) of nursing pigs from 12 herds had pararotavirus antibodies. The physicochemical properties of pararotavirus were examined and compared with those of group A rotaviruses by using the CCIF assay to quantitate in vitro changes in viral infectivity. Pararotavirus was inactivated (greater than or equal to 99% reduction in titer) by heating to 56 degrees C for 30 min, was slightly labile at pH 3 (16 to 34% reduction in titer), and was stable at pH 5 (0 to 17% reduction in titer) and in either (3 to 19% reduction in titer). One group A rotavirus (Gottfried strain) was stable at 56 degrees C (0% reduction in titer), whereas the OSU strain of group A rotavirus was inactivated at this temperature (99% reduction in titer).

Animals↗
PubMed1997

Deletion mapping of functional domains in the rotavirus capsid protein VP6.

VP6, the major capsid protein of rotavirus, oligomerizes into trimers that constitute the intermediate shell of the virions. In order to map functional domains in this protein, we introduced seven internal in-frame deletions within the coding region of gene 6 of human rotavirus strain Wa. Regions of homology among the VP6 proteins of group A and group C rotaviruses were targeted for deletion mutagenesis. The mutant VP6 proteins were expressed in mammalian cells using the recombinant vaccinia virus system and were examined for their ability to oligomerize into trimers as well as to assemble into double-layered virus-like particles upon coexpression with the rotavirus core protein VP2. Deletions that abolished trimerization defined a domain (residues 246 to 314) that maps within a larger region previously found to be critical for oligomerization (amino acids 105 to 328). When the capacity of each mutant to assemble into double-layered virus-like particles was analysed, three different assembly phenotypes were observed. Phenotype I was represented by two deletion mutants lacking residues 246 to 250 and 308 to 314 that produced particles with efficiencies similar to that of wild-type VP6. Phenotype II, characterized by a moderate decrease in the efficiency of particle assembly with respect to that of wild-type VP6, included two mutants with deletions at the C terminus of the protein. Phenotype III was exhibited by three mutants whose abilities to assemble into double-layered virus-like particles were drastically impaired. Two of these mutants define a previously unidentified assembly domain (amino acids 122 to 147) at the N terminus of rotavirus VP6.

Amino Acid Sequence↗
PubMed1997

Detection of group B rotaviruses in fecal samples from diarrheic calves and adult cows and characterization of their VP7 genes.

Groups A, B, and C rotaviruses have been identified in cattle. Group B rotaviruses are associated with sporadic cases of diarrhea in calves and adult cows. From diagnostic submissions to our laboratory, 90 fecal samples from cases of calf diarrhea, 81 fecal samples from cases of adult cow diarrhea (winter dysentery), and 20 fecal samples from case control normal adult cows were tested for group B rotaviruses by polyacrylamide gel electrophoresis (PAGE), and reverse transcription (RT)-PCR (targeting 279 bp of the VP7 gene). In addition, 53 fecal samples from diarrheic adult cows were tested for group B rotaviruses by immune electron microscopy (IEM). By RT-PCR, five samples from calves were group B rotavirus positive (5.6%). Fifteen samples from adult cows with diarrhea were group B rotavirus positive (18.5%), and none of the control fecal samples from normal cows were positive for group B rotaviruses. By PAGE, one calf sample (RT-PCR positive) was group B rotavirus positive (short electropherotype), but none of the adult cow samples were positive for group B rotaviruses. By IEM, 5 (9.4%) of the 53 fecal samples from diarrheic adult cows were group B positive (all were also RT-PCR positive). The VP7 genes of three strains (WD653 from an adult cow and the ATI and Mebus calf strains) were sequenced. The VP7 genes from the three bovine strains showed high (over 90%) nucleotide and deduced amino acid homologies, but lower homologies (48 to 61%) were seen between these genes and the genes from rodent (IDIR) and human (ADRV) group B rotaviruses. Although there were some differences of degree, all inoculated gnotobiotic calves (n = 6) showed abnormal feces between 1 and 3 days after inoculation with each of three strains of group B bovine rotaviruses, and group B rotaviruse, were detected in the feces for up to 2 weeks by RT-PCR but for shorter periods by PAGE or IEM.

Animals↗
PubMedDate not supplied

Atypical rotavirus among diarrhoeic children living in Belém, Brazil.

Atypical rotaviruses were detected in faeces from two diarrhoeic children living in Belém, Pará, Brazil. Rotavirus particles were detected by electron microscopy and the RNA electrophoresis showed patterns which were compatible with group C rotaviruses. Tests for the presence of group A antigen by enzyme-linked-immunosorbent assay (ELISA) were negative. The two children had three successive rotavirus infections and in both cases the atypical strains were excreted at the time of the third infection, causing a mild and short-lasting disease.

Brazil↗
PubMed1996

Serological and genomic characterization of porcine rotaviruses in Thailand: detection of a G10 porcine rotavirus.

A total of 557 fecal specimens collected from piglets with diarrhea in Thailand were examined for rotavirus RNA by polyacrylamide gel electrophoresis. Twenty-three, one, and two samples were positive for group A, group B, and group C rotaviruses, respectively. Two samples exhibited two segments found in picobirnavirus RNA. RNA electropherotyping of 23 group A rotaviruses showed that they were classified into five patterns. By serotyping by enzyme-linked immunosorbent assay and PCR, viruses in 3 and 14 specimens were found to be serotype G3 and serotype G10, respectively. For one specimen, containing a serotype G10 virus (strain P343), virus was isolated in MA-104 cells, and the nucleotide sequences of the VP7 and VP4 genes were determined. Comparative sequence analysis and cross-neutralization tests showed that strain P343 has B223-like G10 and UK-like P7 serotype (or VP4 genotype 5) specificities. Rotaviruses having such antigenic specificities have not been detected in piglets. Thus, the interspecies transmission of rotaviruses between cows and pigs was suggested.

Amino Acid Sequence↗
PubMed1996

Role of enteric pathogens in the aetiology of neonatal diarrhoea in lambs and goat kids in Spain.

Faeces samples from diarrhoeic and non-diarrhoeic lambs and goat kids aged 1-45 days were examined for enteric pathogens. Cryptosporidium parvum was detected in both diarrhoeic lambs (45%) and goat kids (42%) but not in non-diarrhoeic animals. F5+ (K99+) and/or F41+ Escherichia coli strains were isolated from 26% and 22% of the diarrhoeic lambs and goat kids, respectively, although these strains, which did not produce enterotoxins ST I or LT I, were found with similar frequencies in non-diarrhoeic animals. A F5-F41-ST I+ E. coli strain was isolated from a diarrhoeic lamb (0.6%). Verotoxigenic E. coli was isolated from both diarrhoeic and non-diarrhoeic lambs (4.1% and 8.2%, respectively) and there was no association between infection and diarrhoea. The prevalence of group A rotavirus infection in diarrhoeic lambs was very low (2.1%). Groups A and B rotaviruses were detected in three (8.1%) and five (13.5%) diarrhoeic goat kids from two single outbreaks. Group C rotaviruses were detected in four non-diarrhoeic goat kids. An association of diarrhoea and infection was demonstrated only for group B rotavirus. Clostridium perfringens was isolated from 10.8% of the diarrhoeic goat kids but not from non-diarrhoeic goat kids or lambs. Salmonella arizonae was isolated from a diarrhoeic goat kid (2.7%) and the clinical characteristics of the outbreaks where these two latter enteropathogens were found different from the rest. Picobirnaviruses were detected in a diarrhoeic lamb. No coronaviruses were detected using a bovine coronavirus ELISA. No evidence was found of synergistic effect between the agents studied. Enteric pathogens were not found in four (8.7%) and three (20%) outbreaks of diarrhoea in lambs and goat kids, respectively.

Age Factors↗
PubMed1991

Identification of atypical rotaviruses in outbreaks of preweaning and postweaning diarrhea in Québec swine herds.

Intestinal contents of diarrheic pigs from 120 outbreaks of diarrhea were examined for the presence of atypical rotaviruses. Pigs involved in these outbreaks were aged two days to five weeks and samples collected over a period of one year originated from different regions of the province of Québec. Samples were analyzed by polyacrylamide gel electrophoresis (PAGE) for the presence of viral RNA and genome profiles were compared to those of porcine rotavirus A/OSU, B/Ohio and C/Cowden. Based on electropherotypes both typical (group A) and atypical (groups B and C) rotaviruses were identified. Rotaviruses could be demonstrated in 25.8% of outbreaks and together atypical rotaviruses accounted for 46.7% of rotavirus-positive outbreaks. Other common enteropathogens were often present in conjunction with rotaviruses in the preweaning and postweaning outbreaks studied.

Animals↗
PubMed2006

Viral gastroenteritis and genetic characterization of recombinant norovirus circulating in Eastern Russia.

From November 2003 to March 2004 a total of 100 fecal specimens from infants and children with acute gastroenteritis in the city of Birobidzhan, Eastern Russia were tested for the presence of diarrheal viruses by RT-multiplex PCR. Of these, 74 fecal specimens were positive for diarrheal viruses and this represented 74%. Among the diarrheal viruses detected, group A rotavirus was the most prevalent (67%; 67 of 100), followed by norovirus (4%; 4 of 100), group C rotavirus (1%, 1 of 100), sapovirus (1%; 1 of 100), and hepatitis A virus (1%; 1 of 100). It was found that 86.6% (58 of 67) of group A rotavirus were serotyped as G3. Sapovirus and hepatitis A virus were genetically determined to belong to GI/1 and subgenotype 1A, respectively. Interestingly, all norovirus isolates in the study turned out to make a novel cluster when polymerase-based grouping was performed. It is noteworthy to point out that these norovirus isolates were further genetically characterized as naturally occurring recombinants, which were firstly found circulating in the Russian population studied. Breakpoint analysis of recombinant norovirus showed that the recombination site was at the open reading frame (ORF)1/ORF2 overlap. This is the first report of the existence of acute gastroenteritis caused by recombinant norovirus in Eastern Russia.

Acute Disease↗
PubMed1989

Atypical human rotaviruses in the G.D.R.

Following the aetiologic agent of diarrhoea in children aged one to three years we found in the stools a large number of rotaviruses by electron microscopy (EM), although ELISA was negative; the sensitivity of ELISA had been confirmed in previous investigations. In 141 children with diarrhoea 10 conventional (group A) and 21 antigenically distinct rotaviruses were detected. The presence of atypical rotaviruses could be confirmed by electrophoretic analysis of their RNA migration profiles. The electropherotype pattern corresponded to the group C rotaviruses reported by others.

Antigens, Viral↗
PubMed1999

Viral diarrhea in children in Beijing, China.

A study was undertaken from November 1994 to August 1996 to determine the role of viruses in children (< or =5 years of age) hospitalized at Beijing Children Hospital, Beijing China, for acute diarrhea. Stool samples from diarrheal patients were investigated by ELISA, electron microscopy, and RT-PCR for the presence of rotavirus, calicivirus, astrovirus, and adenovirus. Group A rotavirus was detected in 55.9% of all diarrheal patients and comprised 82.5% of all viruses detected. Group A rotavirus samples were further characterized for their G-type specificity by RT-PCR. Four major G types (1-4) were identified. G1 to G4 accounted for 58.9%, 15.7%, 16.8%, and 6.3%, respectively, of the serotyped samples. Almost all rotavirus infections occurred in children less than 1 year of age, with a significant clustering during the winter months. Group C rotavirus was detected in one 18-month-old child. Astroviruses, caliciviruses, and adenoviruses were detected in 8.5%, 7.6%, and 2.5% of the hospitalized children, respectively. This, the first viral etiological study of childhood diarrhea in China, concludes that rotavirus G1-4 strains play an important role in severe diarrhea in Beijing children.

Adenoviridae↗
PubMed2005

Comparison of NSP4 protein between group A and B human rotaviruses: detection of novel diarrhea-causing sequences in group B NSP4.

The human group B rotavirus is a causative agent of severe adult diarrhea. In this study, we analyzed the NSP4 structure of a group B rotavirus strain, CAL-1, and determined whether enterotoxin activity was present in CAL-1 NSP4. CAL-1 NSP4 was comprised of 219 amino acids which was longer than group A and C rotavirus NSP4, and the primary structures of their sequences differed considerably. However, CAL-1 NSP4 had an enterotoxin-like sequence (residues 106-127) that was only 27% identical to the enterotoxin region of NSP4 of KUN (a group A rotavirus strain) at residues 114-135. Interestingly, both of the synthetic peptides, one (residues 99-128) containing the enterotoxin-like sequence and the other (residues 191-219) containing 29 C-terminal amino acids of CAL-1 NSP4, induced diarrhea in 5.5-day-old mice, but not in 17.5-day-old mice, when administered parenterally. Thus, rotavirus "enterotoxin" sequences could be considerably divergent.

Age Factors↗
PubMed2005

Viral diarrhea in Japanese children: results from a one-year epidemiologic study.

A total of 557 fecal specimens from infants and children with acute gastroenteritis in five places (Maizuru, Tokyo, Sapporo, Saga and Osaka) in Japan from July 2002 to June 2003 were tested for the presence of diarrheal viruses by RT-PCR, PRHA, RNA-PAGE and latex agglutination methods. Of these, 56.4% (314) were found positive for diarrheal viruses. Among them, group A rotavirus was the most prevalent (43.6%, 137 of 314) followed by norovirus (29.9%, 94 of 314), adenovirus (7.6%, 24 of 314), group C rotavirus (6.4%, 20 of 314), sapovirus (5.1%, 16 of 314) and astrovirus (1.6%, 5 of 314), respectively. A high rate (7.4%, 19 of 314) of viral mixed infections, including one triple infection (adenovirus, norovirus and astrovirus) was demonstrated. Norovirus infection that usually has a peak during November and January in Japan was detected year-round and highest in September in our study. Norovirus was subjected to molecular genetic analysis by sequencing. The results clearly indicated that norovirus group II was a dominant genogroup (94.3%, 100 of 106). It is noteworthy that noroviruses detected in this study were classified into 8 genotypes (GI/1, GI/4, GII/2, GII/3, GII/4, GII/5, GII/6 and GII/12). Of these, NVGII/4 was the predominant genotype, followed by NVGII/6, and these presented 75.6% (80 of 106) and 11.3% (12 of 106), respectively. Another interesting feature in our study was the sudden appearance and disappearance of SaitamaU16-like strains belonging to NVGII/6 in the short period (January 2003 to June 2003). Our findings confirmed the presence of many diarrheal viruses co-circulating among Japanese infants and children and showed the great genetic diversity among norovirus.

Acute Disease↗