scholarly journals Is Porcine Kobuvirus 1 a Typical Diarrhoeic Pathogen of Piglets?

2017 ◽  
Vol 61 (2) ◽  
pp. 50-55
Author(s):  
H. McFall ◽  
Š. Vilček
Keyword(s):  
Enteric Virus ◽  
Enteric Pathogen ◽  
Porcine Kobuvirus ◽  
Rt Pcr ◽  
Rotavirus A ◽  
Pig Farms ◽  
Rectal Swabs ◽  
Vp6 Gene ◽  
3D Gene

AbstractThe objective of this study was to show if porcine kobuvirus 1 (PKV-1) participates in the development of diarrhoea in piglets. The experiments were focused on comparing the occurrence of PKV-1 with the occurrence of rotavirus A (RVA) infection in suckling pigs on Slovak pig farms. A total of 91 rectal swabs of piglets (age < 28 days) were collected from 8 pig farms. RT-PCR was employed to detect PKV-1 through amplification of the 495 bp fragment of the 3D gene using primers KoVF/ KoVR, and RVA was detected through amplification of the 309 bp fragment of the VP6 gene using primers rot3 and rot5. As expected, the detection of RVA in diarrhoeic piglets was 56.8 % (P < 0.01), while only 14.8 % in healthy animals. These results confirm that RVA is one of the main causes of diarrhoea in young piglets. Comparatively, PKV-1 was detected in approximately equal numbers in the same group of both healthy and diarrhoeic pigs, with 74.1 % in healthy animals and 81.1 % in diarrhoeic animals, which was not statistically significant (P < 0.05). The level of co-infection of both viruses was 11.1 % in healthy animals. A portion of 48.6 % (P < 0.01) of diarrhoeic animals were found with RVA and PKV-1 coinfections. The results of this study indicate that while RVA is an enteric virus, PKV-1 cannot confidently be confirmed as an enteric pathogen.

scholarly journals Diarrhea caused by rotavirus A, B, and C in suckling piglets from southern Brazil: molecular detection and histologic and immunohistochemical characterization

2018 ◽  
Vol 30 (3) ◽  
pp. 370-376 ◽  
Author(s):  
Paula R. Almeida ◽  
Elis Lorenzetti ◽  
Raquel S. Cruz ◽  
Tatiane T. Watanabe ◽  
Priscila Zlotowski ◽  
...  
Keyword(s):  
Virus Detection ◽  
Enteric Virus ◽  
Transmissible Gastroenteritis Virus ◽  
Southern Brazil ◽  
Rt Pcr ◽  
Viral Pathogen ◽  
Rotavirus A ◽  
Diarrhea Virus ◽  
Suckling Piglets ◽  
Transmissible Gastroenteritis

Rotavirus (RV) is an important viral pathogen causing diarrhea in piglets and other mammals worldwide. We describe 34 cases from 4 diarrheal outbreaks caused by RV in unvaccinated farrowing units in southern Brazil from 2011 to 2013. We performed autopsy, histologic examinations, bacterial culture, RV immunohistochemistry (IHC), and enteric virus detection through molecular assays for rotavirus A, B, and C, transmissible gastroenteritis virus, porcine epidemic diarrhea virus, sapovirus, norovirus, and kobuvirus. Histologically, villus atrophy (29 of 34) and epithelial vacuolation (27 of 34) occurred in all 4 outbreaks. Cell debris in the lamina propria occurred in 20 cases, mostly from outbreaks A (8 of 11), C (4 of 6), and D (7 of 11). IHC was positive for RV in 21 of 34 samples. RT-PCR was positive for RV in 20 of 30 samples; RV-C was the most frequently detected RV ( n = 17). Kobuvirus was detected in 11 samples, and, in 3 of them, there was single detection of this enteric virus.

Rotavirus A associated pathology of intestine and mesenteric lymph nodes and occurrence in bovine calves of Gwalior and Bareilly regions

2018 ◽  
Author(s):  
Gayatri Kashyap ◽  
R. Singh ◽  
Y. S. Malik ◽  
R. K. Agrawal ◽  
K. P. Singh ◽  
...  
Keyword(s):  
Small Intestine ◽  
Lymph Nodes ◽  
Mononuclear Cells ◽  
Dairy Farm ◽  
Rt Pcr ◽  
Mesenteric Lymph Nodes ◽  
Fecal Samples ◽  
Rotavirus A ◽  
Mesenteric Lymph ◽  
Vp6 Gene

To understand the pathology of natural cases of rotavirus (RVA) in bovine calves, a total of 40 cases below 6 months died due to diarrhoea were studied, out of which 7 cases (17.5%) turned positive for RVA by RT-PCR. Histopathology of small intestine showed loss of villous enterocytes, blunting and fusion of villi, elongation of crypts and mononuclear cells infiltration in the lamina-propria. The mesenteric lymph nodes were severely depleted of lymphocytes. These changes were corroborated with presence of RVA antigen in sections by dFAT and nucleic acid by RT-PCR. The fluorescent signals were more in mesenteric lymph nodes than in intestine. Besides, 115 rectal fecal samples were also collected from calves for RVA detection by RT-PCR using VP6 gene specific sets of primers. Dead carcasses of calves (n= 40) belonged to organized dairy farm of Bareilly, while rectal fecal samples belonged to both organized (n= 38) and unorganized farms (n= 77) of Bareilly and Gwalior. The overall occurrence of RVA was 19.3% (30/155), comprising 5/37 cases (13.5%) from Gwalior (MP) and 25/118 cases (21.1%) from Bareilly (UP). These findings suggest the infection of RVA widely prevalent in calves and have potential to escape from the intestinal site to mesenteric lymph nodes.

scholarly journals Phylogenetic Study of VP6 Gene of Bovine Rotavirus A and Molecular Survey of Bovine Rotaviruses B and C, and Human G and P Genotypes of Rotavirus A in Calves in Iran

2020 ◽  
Vol 8 (2) ◽  
pp. 66-72
Author(s):  
Ahmad Nazaktabar ◽  
Omid Madadgar
Keyword(s):  
Polyacrylamide Gel Electrophoresis ◽  
Phylogenetic Study ◽  
Rt Pcr ◽  
Japanese Strain ◽  
Rotavirus A ◽  
Bovine Rotavirus ◽  
Study Materials ◽  
Vp6 Gene ◽  
Rotavirus Diarrhea ◽  
Diarrheic Calves

Background: Rotavirus (RV) is one of the most important causes of diarrhea in the calf and human neonates. Rotaviruses are divided into nine different serogroups, of which group A is more important compared to other groups. Objective: This study was performed because of the lack of information about the importance and prevalence of bovine rotaviruses B (RVB) and C (RVC) and human genotypes of rotavirus A (RVA) in the bovine population in Iran. Phylogenetic analysis of VP6 of bovine RVA was the second aim of the present study. Materials and Methods: A total of 581 stool specimens were collected from diarrheic calves of 14 provinces and were analyzed by reverse transcription polymerase chain reaction (RT-PCR) and 485 of them were investigated by PAGE electrophoresis to determine the frequency of three rotaviruses A (RVA), B (RVB), and C (RVC). The presence of human G and P genotypes in Iranian bovine population was also evaluated using semi-nested multiplex RT-PCR. Results: RVA was detected by RT-PCR (VP6 gene detection) in 16.2% (94/581) and by PAGE in 22.16% (108/485) and no positive cases of RVB and RVC were confirmed by polyacrylamide gel electrophoresis (PAGE). This study showed that non-A RV groups (B and C) have little role in calf diarrhea in Iran. The results of the phylogenetic study of VP6 sequences of rotaviruses A identified in this study showed that they all belonged to genotype I2 and were classified into three different branches. Specimen isolated in Zanjan showed the highest difference (maximum identity of 94%) with other sequences and clustered along with the Japanese strain, R22. Human G and P genotypes were not found in the studied samples. Conclusion: The results showed that non-A rotaviruses and human genotypes of RVA are of little importance in calf rotavirus diarrhea in Iran. Also, there is the first phylogenetic study of rotavirus A VP6 protein in Iran.

Enteric viruses in drinking water supplies

2002 ◽  
Vol 2 (3) ◽  
pp. 17-22
Author(s):  
A.P. Wyn-Jones ◽  
J. Watkins ◽  
C. Francis ◽  
M. Laverick ◽  
J. Sellwood
Keyword(s):  
Drinking Water ◽  
Heavy Rainfall ◽  
Liquid Culture ◽  
Plaque Assay ◽  
Enteric Viruses ◽  
Enteric Virus ◽  
The Other ◽  
Raw Water ◽  
Rt Pcr ◽  
Water Supplies

Two rural spring drinking water supplies were studied for their enteric virus levels. In one, serving about 30 dwellings, the water was chlorinated before distribution; in the other, which served a dairy and six dwellings the water was not treated. Samples of treated (40 l) and untreated (20 l) water were taken under normal and heavy rainfall conditions over a six weeks period and concentrated by adsorption/elution and organic flocculation. Infectious enterovirus in concentrates was detected in liquid culture and enumerated by plaque assay, both in BGM cells, and concentrates were also analysed by RT-PCR. Viruses were found in both raw water supplies. Rural supplies need to be analysed for viruses as well as bacterial and protozoan pathogens if the full microbial hazard is to be determined.

Monitoring the marine environment for small round structured viruses (SRSVS): a new approach to combating the transmission of these viruses by molluscan shellfish

1998 ◽  
Vol 38 (12) ◽  
pp. 51-56 ◽  
Author(s):  
K. Henshilwood ◽  
J. Green ◽  
D. N. Lees
Keyword(s):  
Enteric Virus ◽  
Winter Period ◽  
Rt Pcr ◽  
Cloning And Sequencing ◽  
New Approach ◽  
Human Enteric Virus ◽  
Different Strains ◽  
The Uk ◽  
Public Health Protection

This study investigates human enteric virus contamination of a shellfish harvesting area. Samples were analysed over a 14-month period for Small Round Structured Viruses (SRSVs) using a previously developed nested RT-PCR. A clear seasonal difference was observed with the largest numbers of positive samples obtained during the winter period (October to March). This data concurs with the known winter association of gastroenteric illness due to oyster consumption in the UK and also with the majority of the outbreaks associated with shellfish harvested from this area during the study period. RT-PCR positive amplicons were further characterised by cloning and sequencing. Sequence analysis of the positive samples identified eleven SRSV strains, of both Genogroup I and Genogroup II, occurring throughout the study period. Many shellfish samples contained a mixture of strains with a few samples containing up to three different strains with both Genogroups represented. The observed common occurrence of strain mixtures may have implications for the role of shellfish as a vector for dissemination of SRSV strains. These results show that nested RT-PCR can identify SRSV contamination in shellfish harvesting areas. Virus monitoring of shellfish harvesting areas by specialist laboratories using RT-PCR is a possible approach to combating the transmission of SRSVs by molluscan shellfish and could potentially offer significantly enhanced levels of public health protection.

scholarly journals Detection of multiple enteric virus strains within a foodborne outbreak of gastroenteritis: an indication of the source of contamination

2004 ◽  
Vol 133 (1) ◽  
pp. 41-47 ◽  
Author(s):  
C. I. GALLIMORE ◽  
C. PIPKIN ◽  
H. SHRIMPTON ◽  
A. D. GREEN ◽  
Y. PICKFORD ◽  
...  
Keyword(s):  
Arabian Gulf ◽  
Enteric Virus ◽  
Multiple Infections ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Epidemiological Analysis ◽  
Viral Aetiology ◽  
Polymerase Chain ◽  
Most Probable Explanation ◽  
Virus Strains

An outbreak of acute gastroenteritis of suspected viral aetiology occurred in April 2003 in the British Royal Fleet Auxillary ship (RFA) Argus deployed in the Northern Arabian Gulf. There were 37 cases amongst a crew of 400 personnel. Of 13 samples examined from cases amongst the crew, six enteric viruses were detected by reverse transcriptase polymerase chain reaction (RT–PCR). Five different viruses were identified including, three norovirus genotypes, a sapovirus and a rotavirus. No multiple infections were detected. A common food source was implicated in the outbreak and epidemiological analysis showed a statistically significant association with salad as the source of the outbreak, with a relative risk of 3·41 (95% confidence interval of 1·7–6·81) of eating salad on a particular date prior to the onset of symptoms. Faecal contamination of the salad at source was the most probable explanation for the diversity of viruses detected and characterized.

scholarly journals Isolation of Viable SARS-CoV-2 Virus from Feces of an Immunocompromised Patient Suggesting a Possible Fecal Mode of Transmission

2021 ◽  
Vol 10 (12) ◽  
pp. 2696
Author(s):  
Julie Dergham ◽  
Jeremy Delerce ◽  
Marielle Bedotto ◽  
Bernard La Scola ◽  
Valérie Moal
Keyword(s):  
Kidney Transplantation ◽  
Infectious Virus ◽  
Immunocompromised Patient ◽  
Kidney Transplant Recipient ◽  
Enteric Virus ◽  
Rt Pcr ◽  
Virus Particles ◽  
Severe Diarrhea ◽  
Mode Of Transmission ◽  
Compromised Patients

(1) Background: Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) excretion in stools is well documented by RT-PCR, but evidences that stools contain infectious particles are scarce. (2) Methods: After observing a Corona Virus 2019 Disease (COVID-19) epidemic cluster associated with a ruptured sewage pipe, we search for such a viable SARS-CoV-2 particle in stool by inoculating 106 samples from 46 patients. (3) Results: We successfully obtained two isolates from a unique patient with kidney transplantation under immunosuppressive therapy who was admitted for severe diarrhea. (4) Conclusions: This report emphasizes that SARS-CoV-2 is an enteric virus, and infectious virus particles can be isolated from the stool of immune-compromised patients like, in our case, kidney transplant recipient. Immune-compromised patients are likely to have massive multiplication of the virus in the gastrointestinal tract and this report suggests possible fecal transmission of SARS-CoV-2.

Environmental dissemination of group A rotavirus: P-type, G-type and subgroup characterization

2009 ◽  
Vol 60 (3) ◽  
pp. 633-642 ◽  
Author(s):  
F. F. M. Ferreira ◽  
F. R. Guimarães ◽  
T. M. Fumian ◽  
M. Victoria ◽  
C. B. Vieira ◽  
...  
Keyword(s):  
Environmental Samples ◽  
Health Hazard ◽  
Sewage Treatment ◽  
Sewage Treatment Plant ◽  
Treatment Plant ◽  
Northern Region ◽  
Rt Pcr ◽  
Vp6 Gene ◽  
Elution Method ◽  
P Type

Rotaviruses A (RV-A) infection is the most common cause of acute diarrheal diseases in infants and the dissemination of these viruses in the environment represents a public health hazard. The present study aims to evaluate reverse transcription-polymerase chain reaction (RT-PCR) based protocols for the detection of RV-A genes in different types of environmental samples. RV-A were concentrated by the adsorption-elution method using negatively charged membranes associated with a Centriprep Concentrator 50. The RV-A VP4, VP7 and VP6 genes were detected using RT-PCR in river water from the Amazon Hydrographic basin (Northern region) and from wastewater in a sewage treatment plant in Rio de Janeiro (Southeast region), Brazil. RV-A were successfully detected in water environmental samples by the methods used. The detection of the VP6 gene by RT-PCR was the most sensitive for detecting RV-A in environmental samples (44.0%), when compared to the detection of the VP4 (33.3%) and VP7 (25.3%) genes. Based on nucleotide sequence and phylogenetic analysis of the partial VP6 gene, 22 environmental samples were determined to be subgroup II (Wa-like). These results indicate that analysis of environmental samples could possibly make a valuable contribution to studies on the epidemiology of RV-A.

scholarly journals Multiplex Detection of Rotavirus A, B, C, Norovirus GI and GII, Adenovirus, Astrovirus and Sapovirus in a Single PCR System in Stool Samples From Acute Diarrhea Children

2020 ◽  
Author(s):  
Wei Li ◽  
Weiwei Li ◽  
Ran Tao ◽  
Wenqing Xiang ◽  
Mingming Zhou ◽  
...  
Keyword(s):  
High Throughput ◽  
Acute Diarrhea ◽  
Cross Reaction ◽  
Melting Curve Analysis ◽  
Clinical Test ◽  
Rt Pcr ◽  
Rotavirus A ◽  
Diarrhea Virus ◽  
Intestinal Pathogens ◽  
Stool Samples

Abstract Background: Early and accurate identification of infection viruses among children can benefit the personalized medical treatment and management, and reduce the future occurrence of serious symptoms. Thus, it is critical to develop a high-throughput multiplex real-time RT-PCR method to improve the accuracy and efficiency in routine clinical lab tests. Here, we developed a RT-PCR combined with melting curve analysis (RRCMC) method for simultaneous detection of rotavirus A, B, C, norovirus GI and GII, adenovirus, astrovirus and sapovirus. Results: Stool samples were collected from 160 children with acute diarrhea and tested by RRCMC assay. A total of 71 patients were tested positive with norovirus, adenovirus or rotavirus. The RRCMC assay has high specificity. There is no internal cross-reaction through the 8 diarrhea viruses and no cross-reaction of other commonly intestinal pathogens and human genome. The detection limit was ranging from 1×102 to 1×105 copies/ml for each diarrhea virus. Conclusions: In conclusion, the RRCMC method is a suitable rapid clinical test for infection viruses, with the advantages of high-throughput, low cost, high sensitivity, and specificity.

scholarly journals Sensitive SYBR Green—Real Time PCR for the Detection and Quantitation of Avian Rotavirus A

2018 ◽  
Vol 6 (1) ◽  
pp. 2 ◽  
Author(s):  
David De la Torre ◽  
Claudete Astolfi-Ferreira ◽  
Ruy Chacon ◽  
Antonio Piantino Ferreira
Keyword(s):  
Polymerase Chain Reaction ◽  
Real Time ◽  
Sybr Green ◽  
Molecular Techniques ◽  
Economic Losses ◽  
Rt Pcr ◽  
Chain Reaction ◽  
Rotavirus A ◽  
Avian Nephritis Virus ◽  
Polymerase Chain

Avian rotavirus A (ARtV-A) is a virus that affects young birds, causing acute diarrhea and economic losses in the poultry industry worldwide. The techniques used for the diagnosis of ARtV-A include electron microscopy, isolation in cell culture, and serology, as well as molecular techniques, such as the reverse transcription-polymerase chain reaction (RT-PCR). The objective of this work was to standardize a real-time RT-polymerase chain reaction (RT-qPCR) using SYBR Green chemistry for the rapid detection and quantification of ARtV-A from bird tissues and materials fixed on FTA cards on the basis of the nucleotide sequence of segment 6 (S6), which codes for the structural VP6 protein of ARtV-A. The results show the efficient amplification of the proposed target, with a limit of detection (LoD) of one copy gene (CG) per microliter of cDNA and a limit of quantification (LoQ) of 10 CGs per microliter. The efficiency of the primers was determined to be 95.66% using a standard curve, with an R2 value of 0.999 and a slope of −3.43. The specificity was determined using samples coinfected with ARtV-A, the chicken parvovirus, the chicken astrovirus, and the avian nephritis virus as positive controls and commercially available vaccines of the infectious bronchitis virus, infectious bursa disease virus, avian reovirus and healthy organs as negative controls. This technique, which lacks nonspecific PCR products and dimers, demonstrated greater sensitivity and specificity than conventional RT-PCR, and it reduced the analysis time by more than 50%.

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