scholarly journals Porcine rotavirus C in pigs with gastroenteritis on Thai swine farms, 2011–2016

PeerJ ◽  
2018 ◽  
Vol 6 ◽  
pp. e4724 ◽  
Author(s):  
Supansa Tuanthap ◽  
Cherdpong Phupolphan ◽  
Supol Luengyosluechakul ◽  
Ausanee Duang-in ◽  
Apiradee Theamboonlers ◽  
...  
Keyword(s):  
Rt Pcr ◽  
Important Food ◽  
Diarrhea Virus ◽  
Swine Farms ◽  
Glycoprotein G ◽  
Three Samples ◽  
Group A ◽  
Cowden Strain ◽  
P Type ◽  
Porcine Rotavirus

Swine are economically important food animals, but highly contagious porcine epidemic diarrhea virus (PEDV) and rotavirus can afflict pig herds and contribute significantly to piglet morbidity and mortality. While there have been studies on rotavirus group A (RVA) in Thailand, reports of rotavirus group C (RVC) are limited. Here, we aimed to identify the prevalence of RVC circulating on Thai commercial swine farms. We analyzed 769 feces and intestine mucosal contents of pigs affected with diarrhea between 2011 and 2016 using RT-PCR specific for the PEDV spike (S), rotavirus glycoprotein (G) VP7, and protease-sensitive protein (P) VP4 genes. We found that 6.6% (51/769) of samples tested positive for RVC, of which 11 samples were co-infected with RVA and four samples were co-infected with PEDV. Three samples tested positive for all three viruses. Phylogenetic analysis of the VP7 gene showed that the most frequent RVC genotype was G1, which grouped with the prototypic RVC Cowden strain. While G6 and G9 were also common, G3 was relatively rare. Analysis of the VP4 gene revealed that the most common P type was P[5], followed by P[4], P[7], and P[1]. In all, there were six G/P combinations (G6P[5], G1P[1], G1P[4], G1P[5], G9P[4], and G9P[7]), of which G6P[5] was the most predominant.

scholarly journals Molecular characterization of porcine rotavirus C in pigs with gastroenteritis in Thailand, 2011 – 2016

2018 ◽  
Author(s):  
Supansa Tuanthap ◽  
Cherdpong Phupolphan ◽  
Supol Luengyosluechakul ◽  
Usanee Duang-in ◽  
Apiradee Theamboonlers ◽  
...  
Keyword(s):  
Enteric Viruses ◽  
Gene Encoding ◽  
Diarrhea Virus ◽  
Three Samples ◽  
Single Positive ◽  
Group A ◽  
Cowden Strain ◽  
Low Prevalence ◽  
Swine Herds

Swine are economically important food animals, but highly contagious enteric viruses can affect entire swine herds and contribute significantly to piglet morbidity and mortality. The most frequent viruses associated with pig gastroenteritis have been reported as porcine epidemic diarrhea virus (PEDV) and rotavirus. Rotavirus is an important cause of diarrhea in piglets and pigs worldwide, and group A and C types are those that pig herds are mostly affected by. In Thailand, studies on rotavirus group A (RVA) have been reported continuously, whereas information on group C is still limited. In this study, we aimed to identify rotavirus group C (RVC) from the feces and intestinal contents of pigs affected with diarrhea. Seven hundred and sixty-nine samples were collected from swine herds located in difference provinces throughout Thailand. The specimens were tested using virus-specific RT-PCR to detect the gene encoding RVC capsid protein VP7 and VP4. Sequencing analyses showed that 6.6% (51/769) of samples were positive for RVC, one third of which tested as single positive for RVC (34/51). Co-infections with the most frequent enteric viruses, RVA and PEDV were also analyzed. Co-infections of RVA/RVC accounted for 21.6% (11/51) of samples and of PEDV/RVC for 7.8% (4/51) of samples, while three samples (5.9%) tested positive for all three viruses. Infections were not associated with seasonality, since the virus was detected throughout the year. RVC was detected in pigs up to 8 weeks old. Analysis of the partial VP7 gene sequences was suggestive that the predominant genotype was G1, which was closely related to the prototype Cowden strain. Due to P[5] was the most prevalent of VP4 genotype. This study demonstrated the low prevalence of RVC in Thailand, a virus not previously documented in this country.

scholarly journals Molecular characterization of porcine rotavirus C in pigs with gastroenteritis in Thailand, 2011 – 2016

2018 ◽  
Author(s):  
Supansa Tuanthap ◽  
Cherdpong Phupolphan ◽  
Supol Luengyosluechakul ◽  
Usanee Duang-in ◽  
Apiradee Theamboonlers ◽  
...  
Keyword(s):  
Enteric Viruses ◽  
Gene Encoding ◽  
Diarrhea Virus ◽  
Three Samples ◽  
Single Positive ◽  
Group A ◽  
Cowden Strain ◽  
Low Prevalence ◽  
Swine Herds

Swine are economically important food animals, but highly contagious enteric viruses can affect entire swine herds and contribute significantly to piglet morbidity and mortality. The most frequent viruses associated with pig gastroenteritis have been reported as porcine epidemic diarrhea virus (PEDV) and rotavirus. Rotavirus is an important cause of diarrhea in piglets and pigs worldwide, and group A and C types are those that pig herds are mostly affected by. In Thailand, studies on rotavirus group A (RVA) have been reported continuously, whereas information on group C is still limited. In this study, we aimed to identify rotavirus group C (RVC) from the feces and intestinal contents of pigs affected with diarrhea. Seven hundred and sixty-nine samples were collected from swine herds located in difference provinces throughout Thailand. The specimens were tested using virus-specific RT-PCR to detect the gene encoding RVC capsid protein VP7 and VP4. Sequencing analyses showed that 6.6% (51/769) of samples were positive for RVC, one third of which tested as single positive for RVC (34/51). Co-infections with the most frequent enteric viruses, RVA and PEDV were also analyzed. Co-infections of RVA/RVC accounted for 21.6% (11/51) of samples and of PEDV/RVC for 7.8% (4/51) of samples, while three samples (5.9%) tested positive for all three viruses. Infections were not associated with seasonality, since the virus was detected throughout the year. RVC was detected in pigs up to 8 weeks old. Analysis of the partial VP7 gene sequences was suggestive that the predominant genotype was G1, which was closely related to the prototype Cowden strain. Due to P[5] was the most prevalent of VP4 genotype. This study demonstrated the low prevalence of RVC in Thailand, a virus not previously documented in this country.

scholarly journals Circulation of bovine viral diarrhea virus – 1 (BVDV-1) in dairy cattle and buffalo farms in Ismailia Province, Egypt

2015 ◽  
Vol 9 (12) ◽  
pp. 1331-1337 ◽  
Author(s):  
Mohamed Ahmed Soltan ◽  
Rebecca P Wilkes ◽  
Mohamed Nagy Elsheery ◽  
Mahmoud Mohy Elhaig ◽  
Matthhew C Riley ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Dairy Cattle ◽  
Real Time ◽  
Bovine Viral Diarrhea ◽  
Rt Pcr ◽  
Cattle Farm ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Three Samples ◽  
Pcr Products

Introduction: Bovine viral diarrhea (BVD) is one of the most economically significant diseases in the bovine industry causing losses due to diarrhea, reproductive disorders, immunosuppression and mortalities. The aim of our investigation was to detect and subtype BVDV from calves on two dairy cattle and two buffalo farms in Ismailia province, Egypt as an indicator of BVDV infection status in the province. Methodology: A total of 298 blood samples were collected and tested using an optimized one-step, real-time multiplex Taqman-based RT-PCR. All the positive samples by the multiplex real-time RT-PCR were tested using conventional RT-PCR to amplify multiple areas of the genome for further phylogenetic analysis and subtyping. Results: Thirty one (10.4%) of the tested samples were positive for BVDV-1. Only three samples, all from a single dairy cattle farm, had enough viral RNA to be amplified by RT-PCR. The PCR products were sequenced and phylogenetic analysis revealed detection of BVDV-1b. The detected strain is closely related to worldwide BVDV-1b strains, making it difficult to trace its origin. Nucleotide and amino acid alignments of the E2 glycoprotein region of the detected strain with other BVDV-1b strains showed high divergence, with identity ranging from 81.3% to 93.6% and 85.3% to 93.6%, respectively. Conclusion: To our knowledge, this is the first report describing the circulation of BVDV-1b in Egyptian dairy cattle populations.

scholarly journals Genome constellations of 24 porcine rotavirus group A strains circulating on commercial Thai swine farms between 2011 and 2016

PLoS ONE ◽  
2019 ◽  
Vol 14 (1) ◽  
pp. e0211002 ◽  
Author(s):  
Supansa Tuanthap ◽  
Sompong Vongpunsawad ◽  
Supol Luengyosluechakul ◽  
Phanlert Sakkaew ◽  
Apiradee Theamboonlers ◽  
...  
Keyword(s):  
Swine Farms ◽  
Group A ◽  
Porcine Rotavirus

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 Molecular characterization of a rare G1P[19] rotavirus strain from India: evidence of reassortment between human and porcine rotavirus strains

2009 ◽  
Vol 58 (12) ◽  
pp. 1611-1615 ◽  
Author(s):  
Shobha D. Chitambar ◽  
Ritu Arora ◽  
Preeti Chhabra
Keyword(s):  
Amino Acid Sequences ◽  
Rt Pcr ◽  
Coding Regions ◽  
Subgroup Ii ◽  
Human Rotavirus ◽  
Independent Segregation ◽  
Group A ◽  
Porcine Rotavirus ◽  
Porcine Origin

This study pertains to the characterization of a human rotavirus strain (NIV929893) with a rare specificity of G1P[19]. Three structural genes (VP4, VP6 and VP7) and one non-structural gene (NSP4) of strain NIV929893 were subjected to RT-PCR for amplification of entire coding regions. All of the amplicons were sequenced to carry out phylogenetic analysis. The complete amino acid sequences of the VP7 and VP4 gene products showed clustering of the VP7 gene with G1 strains of human origin and the VP4 gene with P[19] strains of porcine origin. The two viral proteins VP6 and NSP4, described previously as genetically linked proteins, were shown to be subgroup II and genotype B of human and porcine origins, respectively. The findings of this study provide evidence of reassortment between VP7/VP6 genes of humans and VP4/NSP4 genes of porcine species and an independent segregation of VP6 and NSP4 genes in a group A human rotavirus strain with G1P[19] specificity.

scholarly journals Possibility for use of RT-PCR technique in establishing presence of bovine viral diarrhea virus in sperm of breeding bulls

2005 ◽  
Vol 59 (3-4) ◽  
pp. 371-381
Author(s):  
Tamas Petrovic ◽  
Sava Lazic ◽  
Milovan Jovicin ◽  
Bosiljka Djuricic
Keyword(s):  
Cell Culture ◽  
Microtiter Plate ◽  
Bovine Viral Diarrhea ◽  
Rt Pcr ◽  
Diarrhea Virus ◽  
Viral Diarrhea ◽  
Culture Cells ◽  
Three Samples ◽  
Pcr Method ◽  
Set Up

The bovine viral diarrhea (BVD) virus is a significant health-economic pathogen in cattle which can be excreted and spread also through sperm of persistently or acutely infected bulls. Native sperm of 6 bulls, found to be negative to the BVD virus by isolating the virus and using the RT-PCR method, was experimentally infected with a tenfold dilution of the non-cytopathogen 22146 strain of the BVD virus with a titer of 105,5. This way, dilutions of the BVD virus from 10-1 to 10-6 (5 x 104 TCID/50 do 0,5 TCID/50 in 0.1 ml native sperm were obtained. From sperm infected in this way, the virus was reisolated on FTB cell culture in a microtiter plate with 96 pools in which each sample of the infected sperm was set up in three samples, and each of them was titrated to a dilution of 1:2 to 1:256. The presence of the BVD virus was proven using the technique of fluorescent antibodies in a second blind passage on FTB culture cells. For cell culture an extremely toxic effect of native sperm to a dilution of 1:64 was established. The BVD virus was reisolated from sperm in all three sperm samples with 5 x 104, 5 x 103 i 5 x 102 TCID/50, and it was not reisolated from sperm with 50, with 5, and with 0.5 TCID/50 BVD virus in 0.1 ml native sperm. At the same time, the presence of the BVD viral genome was proved using the RT-PCR method in the same samples of artificially infected native sperm of bulls. A positive re suit was established in native sperm with 5 x 104, 5 x 103, 5 x 102 and 50 TCID/50 BVD virus n 0.1 ml native sperm. The experiment proved that the RT-PCR method has advantages over the isolation of the BVD virus from samples of native sperm of bulls. These are: shortterm investigations (1 to 2 days) and greater sensitivity (10 times bigger than the isolation of the virus). The isolation of the virus takes at least 10 days, and its greater sensitivity is primarily a result of the cyrotoxic effect of native sperm of bulls on cell culture.

scholarly journals Diarrhea outbreaks in suckling piglets due to rotavirus group C single and mixed (rotavirus groups A and B) infections

2014 ◽  
Vol 34 (5) ◽  
pp. 391-397 ◽  
Author(s):  
Elis Lorenzetti ◽  
Danilo T. Stipp ◽  
Flávia Possatti ◽  
Joice E.T. Campanha ◽  
Alice F. Alfieri ◽  
...  
Keyword(s):  
Sequence Analysis ◽  
Single Infection ◽  
Mixed Infections ◽  
Rt Pcr ◽  
Fecal Samples ◽  
Neonatal Diarrhea ◽  
The Third ◽  
Group A ◽  
Suckling Piglets ◽  
Porcine Rotavirus

Porcine group A rotavirus (PoRVA) is a major cause of neonatal diarrhea in suckling and recently weaned piglets worldwide. The involvement of non-group A rotavirus in cases of neonatal diarrhea in piglets are sporadic. In Brazil there are no reports of the porcine rotavirus group C (PoRVC) as etiologic agent of the diarrhea outbreaks in piglets. The aim of this study was to describe the identification of rotavirus group C in single and in mixed infection with rotavirus groups A and B in three neonatal diarrhea outbreaks in suckling (<21-day-old) piglets, with 70% to 80% and 20% to 25% of morbidity and lethality rates, respectively, in three pig herds located in the state of Santa Catarina, Brazil. The diagnosis of PoRV in the diarrheic fecal samples was performed using polyacrylamide gel electrophoresis (PAGE) to identify the presence of porcine rotavirus groups A, B (PoRVB), and C, and by RT-PCR (PoRVA and PoRVC) and semi-nested (SN)-PCR (PoRVB) to partially amplify the VP4 (VP8*)-VP7, NSP2, and VP6 genes of PoRVA, PoRVB, and PoRVC, respectively. One RT-PCR (PoRVA and PoRVC) and SN-PCR (PoRVB) product of each group of rotavirus of each diarrhea outbreak was submitted to nucleotide (nt) sequence analysis. Based on the PAGE technique, 4 (25%) and 1 (6.25%) of the 16 diarrheic fecal samples evaluated in the first outbreak presented PoRVA and PoRVC electropherotype, respectively, and 11 (68.75%) were negative. In the second outbreak, 3 (42.85%) of the 7 fecal samples evaluated presented PoRVA electropherotype, and in 3 (42.85%) and in 1 (14.3%) fecal samples were detected inconclusive and negative results, respectively. Three (30%) of the 10 fecal samples of the third outbreak presented PoRVC electropherotype; 5 (50%) and 2 (20%) samples showed negative and inconclusive results, respectively. Based on the RT-PCR and SN-PCR assays in the first neonatal diarrhea outbreak, PoRVC was detected in 13 (81.2%) of the 16 diarrheic fecal samples evaluated. PoRVC single infection was identified in 4 (25%) of these samples and mixed infections with PoRVA and PoRVB in 9 (56.2%) fecal samples. All of the seven diarrheic fecal samples evaluated from the second neonatal diarrhea outbreak were positive for PoRVC, whereas its mixed infection with other PoRV groups was detected in 4 (57.2%) samples. In the third outbreak, PoRVC in single infection was detected in all of the 10 diarrheic fecal samples analyzed. In the nt sequence analysis, the PoRVA strains of the first and second outbreaks demonstrated higher nt identity with G4P[6] and G9P[23] genotypes, respectively. The PoRVB strains (first and second outbreaks) and the PoRVC strains (first, second, and third outbreaks) showed higher nt identity and clustered in the phylogenetic tree with PoRVB and PoRVC strains that belong to the N4 and I1 genotypes, respectively. This is the first description in Brazil of the involvement of PoRVC in the etiology of diarrhea outbreaks in suckling piglets. The results of this study demonstrated that PoRVC, in both single and mixed infections, is an important enteropathogen involved in neonatal diarrhea outbreaks in piglets and that the use of more sensitive diagnostic techniques allows the identification of mixed infections involving two or even three groups of PoRV, which may be more common than previously reported.

scholarly journals VP7 and VP4 Genotyping of Human Group A Rotavirus in Buenos Aires, Argentina

2000 ◽  
Vol 38 (1) ◽  
pp. 252-259
Author(s):  
M. H. Argüelles ◽  
G. A. Villegas ◽  
A. Castello ◽  
A. Abrami ◽  
P. D. Ghiringhelli ◽  
...  
Keyword(s):  
Multiplex Pcr ◽  
Buenos Aires ◽  
Polyacrylamide Gel Electrophoresis ◽  
Immunoglobulin M ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rt Pcr ◽  
Group A Rotavirus ◽  
Iga Antibodies ◽  
Group A ◽  
P Type

ABSTRACT Specific and sensitive tests for the detection and typing of group A rotavirus strains are needed for a more comprehensive knowledge of the epidemiology of rotaviral infection. In this study 500 stool specimens taken from 1996 to 1998 from children with acute diarrhea in Buenos Aires were examined. Group A rotavirus was unequivocally demonstrated in 62% of the samples tested by enzyme-linked immunosorbent assay (ELISA) for detection of VP6 antigen, polyacrylamide gel electrophoresis of double-stranded RNA, and reverse transcription-PCR (RT-PCR) for amplification of the VP7:G (1,062 bp) and VP4:P (876 bp) genes. Only five positive specimens were found by RT-PCR but not by ELISA. G and P typing was carried out by nested amplification of variable sequences of the VP7 and the VP4 genes with six G- and five P-type-specific primers (multiplex PCR). Results obtained by this method showed the prevalence of the following G and P types: G1, 39%; G2, 43%; G4, 4%; P[8], 16%; P[4], 71%. Unexpectedly, the G-P type combination most frequently found was G2P[4] (43%) rather than G1P[8] (12%), which is the most commonly found worldwide. Unusual strains of the type G1P[4] accounted for 14% of the total, while mixed infections with more than one type were found in 10% of the samples. Detection of fecal rotavirus-specific immunoglobulin M (IgM) and IgA antibodies in consecutive samples of two patients taken at daily intervals demonstrated that high levels of IgM and IgA antibodies were detected on day 1 after the onset of disease and that the samples remained positive for about 10 days, after which virus shedding was no longer observed. Multiplex PCR offers a sensitive and specific alternative to determine the prevalence of group A rotavirus G and P types and to identify the emergence of uncommon strains, whereas detection of fecal IgM and IgA antibodies represents a useful supplement to virus detection for the diagnosis of current or recently acquired infections.

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