scholarly journals Dual Infection of Gnotobiotic Calves with Bovine Strains of Group A and Porcine-Like Group C Rotaviruses Influences Pathogenesis of the Group C Rotavirus

1999 ◽  
Vol 73 (11) ◽  
pp. 9284-9293 ◽  
Author(s):  
K. O. Chang ◽  
P. R. Nielsen ◽  
L. A. Ward ◽  
L. J. Saif
Keyword(s):  
Villous Atrophy ◽  
Polyacrylamide Gel Electrophoresis ◽  
Dual Infection ◽  
The United States ◽  
Rt Pcr ◽  
Fecal Samples ◽  
Group C Rotavirus ◽  
Bovine Strain ◽  
Group A Rotaviruses ◽  
Group A

ABSTRACT There is serological evidence that bovine group C rotaviruses exist in the United States, but there are no reports of their isolation. Ninety fecal samples from calves with diarrhea, 81 samples from adult cows with diarrhea (winter dysentery), and 20 fecal samples from healthy adult cows were tested for group C rotaviruses by polyacrylamide gel electrophoresis, immune electron microscopy, and reverse transcription-PCR (RT-PCR). Three samples from adult cow diarrhea cases were positive only by RT-PCR, and a group C rotavirus was isolated from a positive sample in monkey kidney (MA104) cells (WD534tc/C). Genetically and serologically, the WD534tc/C strain was more closely related to the Cowden porcine group C strain than to the Shintoku bovine strain. Because the original cow feces also contained a group A rotavirus (detected after passage in cell culture), we hypothesized that such dual-rotavirus infections might play a role in the pathogenesis and host adaptation of rotaviruses. Thus, we examined the pathogenesis of WD534tc/C alone or combined with virulent (IND/A) or attenuated (NCDV/A) bovine group A rotaviruses in gnotobiotic calves. WD534tc/C alone induced diarrhea without (or with limited) virus shedding in inoculated calves (n = 3). In contrast, all calves coinfected with WD534tc/C and IND/A (n = 2) developed diarrhea and shed both viruses, whereas calves coinfected with WD534tc/C and NCDV/A (n= 3) developed diarrhea but did not shed either virus. Infection with WD534tc/C or NCDV/A alone caused only mild villous atrophy (jejunum and/or ileum), whereas dual infection with both viruses induced lesions throughout the small intestine. Although IND/A alone caused villous atrophy, more-widespread small intestinal lesions occurred in calves coinfected with WD534tc/C and IND/A. In conclusion, coinfection of calves with group A rotaviruses enhanced fecal shedding of a bovine group C rotavirus and the extent of histopathological lesions in the small intestines. Thus, our findings suggest a potential novel hypothesis involving dual infections for the adaptation of heterologous rotaviruses to new host species.

scholarly journals Characterization of Group C Rotaviruses Associated with Diarrhea Outbreaks in Feeder Pigs

1999 ◽  
Vol 37 (5) ◽  
pp. 1484-1488 ◽  
Author(s):  
Yunjeong Kim ◽  
Kyeong-Ok Chang ◽  
Barbara Straw ◽  
Linda J. Saif
Keyword(s):  
Amino Acid ◽  
Clinical Signs ◽  
Watery Diarrhea ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Fecal Samples ◽  
Group C Rotavirus ◽  
Group A Rotaviruses ◽  
Group A ◽  
Cowden Strain

Feces and serum specimens were collected from three farms in Michigan on which ∼50-lb (8- to 9-week-old) pigs experienced diarrhea just after placement into all-in-all-out finishing barns. The clinical signs (profuse watery diarrhea lasting about 2 weeks and no vomiting) were similar on all farms, and the morbidity rate was high (ranging from 60 to 80%) but without mortality. Eleven diarrheic fecal samples from the farms were tested for group A and C rotaviruses by immune electron microscopy (IEM) and various assays. IEM indicated that the fecal samples reacted only with antiserum against group C rotaviruses, and polyacrylamide gel electrophoresis indicated that the samples had characteristic genomic electropherotypes for group C rotavirus. Group C rotavirus was detected by cell culture immunofluorescence (CCIF) tests in nine fecal samples, but no group A rotavirus was detected by enzyme-linked immunosorbent assay or CCIF. By reverse transcription (RT)-PCR, all 11 fecal samples were positive for group C rotaviruses, with only 2 samples positive for group A rotaviruses. However, a second amplification of RT-PCR products using nested primers detected group A rotaviruses in all samples. Analysis of nucleotide and deduced amino acid sequences of the RT-PCR product (partial-length VP7) of the group C rotavirus showed 87.2 to 91% nucleotide identity and 92.6 to 95.9% amino acid identity among two strong samples from the different farms and the Cowden strain of porcine group C rotavirus. All nine convalescent-phase serum samples tested had neutralizing antibodies to the Cowden strain, and the majority of them had neutralizing antibody against group A rotaviruses (OSU or/and Gottfried strains) by fluorescent focus neutralization tests. Although group C rotaviruses have been reported as a cause of sporadic diarrhea in suckling or weanling pigs, to our knowledge, this is the first report of epidemic diarrhea outbreaks associated with group C rotavirus in older pigs.

scholarly journals Electropherotypes and G-Types of Group A Rotaviruses Detected in Children with Diarrhea in Lagos, Nigeria

ISRN Virology ◽  
2013 ◽  
Vol 2013 ◽  
pp. 1-5 ◽  
Author(s):  
Christianah Idowu Ayolabi ◽  
David Ajiboye Ojo ◽  
George Enyimah Armah
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Migration Pattern ◽  
Genomic Diversity ◽  
Rt Pcr ◽  
Group A Rotaviruses ◽  
Group A ◽  
Stool Samples ◽  
Health Care Centers ◽  
Virus Isolates

Approximately over 500,000 children die annually due to severe dehydrating diarrhea caused by rotaviruses. This work investigated rotavirus infection among children less than 5 years with diarrhea in Lagos and determined the circulating electropherotypes and genotypes of the virus isolates. Three hundred and two (n=302) stool samples from children below 60 months were collected from different hospitals and health care centers in Lagos and subjected to enzyme immunoassay (EIA) to determine the presence of Group A rotavirus, RT-PCR to determine the G-types, and polyacrylamide gel electrophoresis (PAGE) to determine the electropherotypes. The results show that 60.3% of the samples showed distinct rotavirus RNA migration pattern, having long electropherotypes (55.3%) of seven variations dominating over the short electropherotypes (44.5%). Six different G-types were detected (G1, G2, G3, G4, G9, and G12). Serotypes G1 and G12 showed long electropherotypic pattern while G2, G3, and G9 exhibited either short or long electropherotype. All G4 detected show short electropherotypic pattern. In conclusion, information on the genomic diversity and RNA electropherotypes of rotaviruses detected in children with diarrhea in Lagos is reported in this study.

scholarly journals Enzyme-Linked Immunosorbent Assay Based on Recombinant Human Group C Rotavirus Inner Capsid Protein (VP6) To Detect Human Group C Rotaviruses in Fecal Samples

1998 ◽  
Vol 36 (11) ◽  
pp. 3178-3181 ◽  
Author(s):  
V. L. A. James ◽  
P. R. Lambden ◽  
E. O. Caul ◽  
, and I. N. Clarke
Keyword(s):  
Immunosorbent Assay ◽  
Polyacrylamide Gel Electrophoresis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Human Group ◽  
Fecal Samples ◽  
Reverse Transcription Pcr ◽  
The United Kingdom ◽  
Group C Rotavirus ◽  
Group A ◽  
Rotavirus Infections

A recent study showed that 43% of a population in the United Kingdom were seropositive for group C rotavirus. The higher than expected incidence may be due to limited diagnosis of acute human group C rotavirus infections because no routine test is available. Human group C rotavirus infections are routinely diagnosed by electron microscopy (EM) and a negative group A rotavirus enzyme-linked immunosorbent assay (ELISA) result. An antigen-detection ELISA was developed with hyperimmune antibodies raised to human group C rotavirus recombinant VP6 (Bristol strain) expressed in insect cells. The assay was used to screen fecal samples to determine the prevalence of group C rotavirus infection. Samples positive by ELISA were confirmed by EM, polyacrylamide gel electrophoresis of double-stranded RNA, or detection of the VP6 gene by reverse transcription-PCR. Retrospective analysis indicated a 1 to 2% detection rate of positivity among samples from patients with acute diarrhea.

scholarly journals Isolation, immunochemical demonstration of field strains of porcine group A rotaviruses and electrophoretic analysis of RNA segments of group A and C rotaviruses

2012 ◽  
Vol 51 (No. 5) ◽  
pp. 288-295 ◽  
Author(s):  
R. Smitalova ◽  
L. Rodak ◽  
I. Psikal ◽  
B. Smid
Keyword(s):  
Cell Line ◽  
Electrophoretic Analysis ◽  
Elisa Test ◽  
Farm Animals ◽  
Economic Losses ◽  
Rt Pcr ◽  
Group C Rotavirus ◽  
Group A Rotaviruses ◽  
Group A ◽  
Virus Isolates

Rotaviruses are major cause of acute diarrhea in animals and humans which can result in huge economic losses in farm animals including pigs. We collected 195 samples of feces of diarrhoeic animals. Rotavirus was demonstrated by electron microscopy using the method of negative staining in 27 samples and by ELISA test using monoclonal antibodies to the group antigen VP6 in 44 samples. Nine samples were selected for virus isolation. Three virus isolates (P375/4, P410/4 and P646/1) were successfully adapted to growth in cell line MA-104. These isolates were allocated to group A rotaviruses based on ELISA, immunoperoxidase test and electropherotype analysis. Electropherotype analysis demonstrated changes during passage in cell line in two of the three isolates. The selected sample P543/1 proved negative in ELISA in a fecal sample. Electropherotype analysis of this sample revealed a “longer” electropherotype profile. The profile was suggestive of group C rotavirus. Rotavirus group C was confirmed by RT-PCR and by sequence analysis in this sample.

scholarly journals Gastroenteric virus detection in fecal samples from women in Goiânia, State of Goiás, Brazil

2010 ◽  
Vol 43 (3) ◽  
pp. 240-243 ◽  
Author(s):  
Rui Gilberto Ferreira ◽  
Ana Maria Tavares Borges ◽  
Fabiola Souza Fiaccadori ◽  
Menira Borges de Lima Dias e Souza ◽  
Rodrigo Alessandro Togo Santos ◽  
...  
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Virus Detection ◽  
Hiv Positive ◽  
Risk Of Infection ◽  
Rt Pcr ◽  
Fecal Samples ◽  
Hiv Positive Women ◽  
A Prospective Study ◽  
Immunoenzymatic Assays ◽  
Hiv Seropositive

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

scholarly journals Group B rotaviruses similar to strain CAL-1, have been circulating in Western India since 1993

2004 ◽  
Vol 132 (4) ◽  
pp. 745-749 ◽  
Author(s):  
S. D. KELKAR ◽  
J. K. ZADE
Keyword(s):  
Severe Disease ◽  
Polyacrylamide Gel Electrophoresis ◽  
Epidemiological Studies ◽  
Western India ◽  
Human Group ◽  
Older Children ◽  
Pcr Products ◽  
Group A Rotaviruses ◽  
Group A ◽  
Group B

Generally, group A rotaviruses are the most common cause of paediatric diarrhoea. However, group B rotavirus, adult diarrhoea rotavirus (ADRV), was found to be involved in epidemics of severe gastroenteritis in several areas of China during 1982–1983 and had resulted in more than one million cases among adults as well as older children. Human group B rotavirus has been rarely reported outside China, but has been detected first from five adults with diarrhoea in Kolkata, India during 1997–1998 (strain CAL-1). During epidemiological studies at the National Institute of Virology (NIV) on hospitalized diarrhoea patients at Pune, India, faecal specimens from patients of >5 years age, which were negative for group A rotavirus by ELISA were tested by polyacrylamide gel electrophoresis (PAGE). We detected rotavirus RNA migration patterns similar to that of group B rotavirus in three faecal specimens from adults, two from the specimens collected in 1993 and one in 1998 from sporadic diarrhoea cases. RT–PCR was carried out using primers derived from gene 8 which codes for the NS2 protein, followed by nested PCR, which confirmed the presence of group B rotavirus in all three specimens. The sequences of the PCR products of NIV specimens were compared with that of CAL-1, ADRV and IDIR (infectious diarrhoea of infant rat) belonging to group B rotaviruses. The sequence analysis of the PCR products showed the highest identity with CAL-1, which was reported from Kolkata, India during 1997–1998. The finding suggests that human group B rotaviruses have been circulating in Pune, India, since 1993. This emerging virus may lead to more severe disease among adults in India. There is a need for surveillance of group B rotavirus infections, especially in adult diarrhoea cases and seroepidemiological studies on group B rotavirus are required among humans and animals of Western Maharashtra, India.

scholarly journals Molecular Detection of Rotavirus in Mollusks from the Oued El Maleh Estuary of Mohammedia, Morocco

2021 ◽  
Author(s):  
Abderrahim Hatib ◽  
Najwa Hassou ◽  
Abdelouahab Benani ◽  
Jamal Eddine Hafid ◽  
Moulay Mustapha Ennaji
Keyword(s):  
Real Time ◽  
Enteric Bacteria ◽  
Human Populations ◽  
Rt Pcr ◽  
Wet Season ◽  
Rotavirus A ◽  
Viral Loads ◽  
Wide Range ◽  
Group A Rotaviruses ◽  
Group A

Viral outbreaks can result from the consumption of contaminated bivalve mollusks. However, despite the regulation related to enteric bacteria in food products, the consumption of raw and undercooked mollusks remains linked to viral epidemics in human populations. Real-time RT-PCR is a highly sensitive approach for detecting and quantifying enteric viruses, and after eliminating enzymatic amplification inhibitors from samples of interest, sensitive and specific tests, like real-time RT-PCR, can facilitate the detection and quantification of a wide range of viruses that are concentrated in mollusk digestive tissues. The aim of the present study was to evaluate the prevalence of Group-A rotaviruses in mussel (Mytilus edulis Linnaeus, 1758) specimens (n=576) collected downstream of the Oued El Maleh Estuary, which is along the coast of Mohammedia City in Morocco, using real-time RT-PCR. Rotavirus A RNA was detected in 37.5% (n=18) of the 48 sample batches, and viral loads ranged from 0.42×101 to 1.8603×104 genomic copies per g digestive tissue. Most (72.22%) of the positive samples were collected during the wet season (September-April), and the probability of detecting rotaviruses was significantly greater during the wet season than during the dry season (P<0.001). Monitoring Rotavirus A and similar viruses in shellfish may help prevent viral contamination and preserve public health.

scholarly journals Rotavirus genotyping in gastroenteritis cases of an infantile population from Western Brazilian Amazonia

2012 ◽  
Vol 45 (4) ◽  
pp. 520-522 ◽  
Author(s):  
Maria Sandra Moura Costa ◽  
Paulo Afonso Nogueira ◽  
Gleicienne Félix Magalhães ◽  
Paula Taquita ◽  
Luis André Mariúba ◽  
...  
Keyword(s):  
Molecular Characterization ◽  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Brazilian Amazon ◽  
Rt Pcr ◽  
Rotavirus A ◽  
Positive Stool ◽  
Group A ◽  
Stool Samples

INTRODUCTION: During the period from 2000 to 2002, 79 rotavirus-positive stool samples were collected from children presenting diarrhea in the Western Brazilian Amazon. METHODS: Molecular characterization of the G and P genotypes was performed using RT-PCR and electropherotyping analysis by polyacrylamide gel electrophoresis. RESULTS: A total of 59 samples were confirmed as group A rotavirus. A long electrophoretic profile was exhibited by the G1P[8], G3P[8], and G4P[8] genotypes. The G1P[8] genotype was found in greater proportion. The short electropherotype was exhibited only by G2 genotype strains. CONCLUSIONS: The proportion of the rotavirus genotypes observed was not different from that in other areas of Brazil. This study is the first genotyping of rotavirus in the Western Brazilian Amazon.

scholarly journals Transfection of exogenous rotavirus rearranged RNA segments in cells infected with a WT rotavirus results in subsequent gene rearrangements

2014 ◽  
Vol 95 (9) ◽  
pp. 2089-2098 ◽  
Author(s):  
Sarah Duponchel ◽  
Cécile Troupin ◽  
Lan Trang Vu ◽  
Aurélie Schnuriger ◽  
Germain Trugnan ◽  
...  
Keyword(s):  
Cell Culture ◽  
Acute Gastroenteritis ◽  
Reverse Genetics ◽  
Gene Rearrangements ◽  
Recombinant Viruses ◽  
Bovine Strain ◽  
The Family ◽  
Viral Progeny ◽  
Group A Rotaviruses ◽  
Group A

Group A rotaviruses, members of the family Reoviridae, are a major cause of infantile acute gastroenteritis. The rotavirus genome consists of 11 dsRNA segments. In some cases, an RNA segment is replaced by a rearranged RNA segment, which is derived from its standard counterpart by partial sequence duplication. It has been shown that some rearranged segments are preferentially encapsidated into viral progenies after serial passages in cell culture. Based on this characteristic, a reverse genetics system was used previously to introduce exogenous segment 7 rearrangements into an infectious rotavirus. This study extends this reverse genetics system to RNA segments 5 and 11. Transfection of exogenous rotavirus rearranged RNA segment 5 or 11 into cells infected with a WT helper rotavirus (bovine strain RF) resulted in subsequent gene rearrangements in the viral progeny. Whilst recombinant viruses were rescued with an exogenous rearranged segment 11, the exogenous segment was modified by a secondary rearrangement. The occurrence of spontaneous rearrangements of WT or exogenous segments is a major hindrance to the use of this reverse genetics approach.

scholarly journals MOLECULAR EPIDEMIOLOGICAL STUDIES ON ROTAVIRUS INFECTION CAUSING SEVERE DIARRHEA IN HUMAN, ANIMALS AND POULTRY

2013 ◽  
Vol 9 (2) ◽  
pp. 167-175 ◽  
Author(s):  
MN Alam ◽  
MM Alam ◽  
A Nahar ◽  
N Kobayashi
Keyword(s):  
Gel Electrophoresis ◽  
Polyacrylamide Gel Electrophoresis ◽  
Winter Season ◽  
Epidemiological Studies ◽  
Polyacrylamide Gel ◽  
Fecal Samples ◽  
Rotavirus Infection ◽  
Severe Diarrhea ◽  
Human Rotavirus ◽  
Group A

The epidemiology of rotavirus infection in human, calves, sheep, goats and poultry were studied. Among total  of 800 collected fecal samples , 320 samples from human, 125 samples from calves, 82 samples from sheep, 7 samples from goats, 267 samples from poultry were collected from July 2010 to May 2011 and examined by Polyacrylamide Gel Electrophoresis and Silver Staining (PAGE-SS) technique for the detection of presence of rotavirus dsRNA. Human rotavirus was detected 10.94 % (35/320) in diarrhoeic fecal samples. The highest prevalence was recorded in September 33.33% and the lowest in May 4.54%. The prevalence of rotavirus infections was 33.33% in autumn (September), 11.69% in late autumn (October-November), 9.6% in winter (December-January), 9.72% in spring (February- March), 6.12% in summer (April-May) season in diarrhoeic samples indicated the presence of rotavirus in human round the year in Bangladesh and as such no marked seasonal variation in rotavirus infection in human. No calves, sheep and goat fecal sample was found positive for rotavirus on PAGE-SS technique. During the study period, 267 faecal samples (diarrhoeic and nondiarrhoeic) of chicken (from one day to one month of age) were tested and only one was found positive on PAGE-SS technique for rotavirus infection (0.38%; 1/267). The positive cases were found in samples collected in winter season from layer chicks aged 10 days. The migration patterns of detected positive strains were not similar on polyacrylamide gel electrophoresis and their migration speed was different types. Five electropherotypes were determined among 35 human rotavirus positive samples. All the electropherotypes were under group A and long pattern. The genome migration of avian rotavirus was distinct from human types and under group D and long pattern. In the present study, it was not investigated that bacteria, parasite or any other viruses which might be responsible for development of diarrhoea.DOI: http://dx.doi.org/10.3329/bjvm.v9i2.13473

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