Comparison between oral fluid samples and pooled serum samples for the detection of antibodies against Porcine Reproductive and Respiratory Syndrome Virus in weaning pig herds

2020 ◽  
Vol 162 (9) ◽  
pp. 531-538
Author(s):  
R Graage ◽  
S Beck ◽  
M Koch ◽  
M Dolezal ◽  
L Schwarz ◽  
...  
Keyword(s):  
Oral Fluid ◽  
Respiratory Syndrome Virus ◽  
Serum Samples ◽  
Weaning Pig

scholarly journals Molecular detection of Porcine Reproductive and Respiratory Syndrome Virus, Porcine Circovirus 2 and Hepatitis E Virus in oral fluid compared to their detection in faeces and serum.

2020 ◽  
Author(s):  
Jan Plut ◽  
Urska Jamnikar-Ciglenecki ◽  
Marina Stukelj
Keyword(s):  
Hepatitis E Virus ◽  
Pathogen Detection ◽  
Oral Fluid ◽  
Hepatitis E ◽  
Viral Disease ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus ◽  
Respiratory Syndrome Virus ◽  
Rt Pcr ◽  
Serum Samples

Abstract Background: Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), Porcine Circovirus Type 2 (PCV2) and Hepatitis E virus (HEV) are common and economically important viral disease causative agents detected in pig oral fluid (OF), faeces and serum at some infection stages. The purpose of this study was to detect PRRSV, PCV2 and HEV on six pig farms to determine which of the three sample types, OF, faeces or serum is appropriate for the diagnosis of these viruses in different pig categories.The following pig categories were included: 5 weeks-old (w/o), 7 w/o, 9 w/o, 11 w/o weaners, fatteners and breeding sows. Pursuant to the preliminary detection of each pathogen at the selected farms, OF samples, faeces, serum pools and 10 individual sera were examined, using PCR, for each age category. If any of the viruses were found in pools of faeces and OF, then faeces and OF from positive farms were tested separately for each pig category. The viral nucleic acids were detected using RT-PCR, PCR and real-time RT-PCR, for PRRSV, PCV2 and HEV respectively.Results: PRRSV and HEV were detected on one farm and PCV2 on three others, positive results being more often obtained from the OF than from the faeces of the same animals. Ten individual serum samples from pigs from the same group of animals were also tested. The viruses were detected in almost all individual sera and OF in the same pig category with some exceptions: PRRSV was detected in the OF of fatteners but was absent in their sera; on Farm 2, PCV2 was detected in sera of 11 w/o pigs and fatteners but absent in group samples of their OF and, vice versa, in case of 9 w/o animals; HEV was detected in the OF of the youngest, 5 w/o weaners and absent in sera of the same age group.Conclusions: The primary finding of the study is that OF is a welfare-friendly, non-invasive and highly efficient matrix for pathogen detection, thus evidencing the usefulness of pig OF as a matrix in which each of the three viruses considered can be detected with the highest probability.

scholarly journals Development and Validation of an Assay To Detect Porcine Reproductive and Respiratory Syndrome Virus-Specific Neutralizing Antibody Titers in Pig Oral Fluid Samples

2013 ◽  
Vol 20 (8) ◽  
pp. 1305-1313 ◽  
Author(s):  
Kang Ouyang ◽  
Basavaraj Binjawadagi ◽  
Apisit Kittawornrat ◽  
Chris Olsen ◽  
Jagadish Hiremath ◽  
...  
Keyword(s):  
Disease Surveillance ◽  
Neutralizing Antibodies ◽  
Oral Fluid ◽  
Low Cost ◽  
Herd Immunity ◽  
Neutralizing Antibody ◽  
Respiratory Syndrome Virus ◽  
Serum Samples ◽  
Antibody Titers ◽  
Breeding Farms

ABSTRACTPorcine reproductive and respiratory syndrome virus (PRRSV)-specific neutralizing antibodies (NA) are important for clearing the virus. Pen-based pig oral fluid samples for disease surveillance are gaining in importance due to the ease of collection and low cost. The aim of this study was to develop a PRRSV-specific NA assay to determine NA titers in pig oral fluid samples. At first, we standardized the PRRSV NA assay using pen-based pig oral fluid samples collected over a period of 3 months from a herd of swine that received a PRRSV modified live vaccine (PRRS-MLV), and we also used oral fluid and serum samples collected from individual boars that were vaccinated with PRRS-MLV or infected with a virulent PRRSV strain. Our results suggest that a PRRSV NA titer of >8 in oral fluid samples is virus specific and can be detected beginning at 28 days after vaccination or infection. To validate the assay, we used 104 pen-based pig oral fluid and five representative serum samples from each pen of unknown history, as well as 100 serum samples from repeatedly vaccinated sows and oral fluid samples of their respective litters belonging to four different swine-breeding farms. Our results demonstrated that PRRSV NA titers in oral fluid samples are correlated with serum sample titers, and maternally derived PRRSV-specific NA titers could be detected in litters at the time of weaning. In conclusion, we have standardized and validated the pig oral fluid-based PRRSV NA assay, which has 94.3% specificity and 90.5% repeatability. The assay can be used to monitor herd immunity against PRRSV in vaccinated and infected herds of swine.

scholarly journals Development of a Fluorescent Microsphere Immunoassay for Detection of Antibodies against Porcine Reproductive and Respiratory Syndrome Virus Using Oral Fluid Samples as an Alternative to Serum-Based Assays

2011 ◽  
Vol 19 (2) ◽  
pp. 180-189 ◽  
Author(s):  
Robert J. Langenhorst ◽  
Steven Lawson ◽  
Apisit Kittawornrat ◽  
Jeffrey J. Zimmerman ◽  
Zhi Sun ◽  
...  
Keyword(s):  
Antibody Response ◽  
Disease Surveillance ◽  
Oral Fluid ◽  
Nonstructural Protein ◽  
Respiratory Syndrome Virus ◽  
Type I ◽  
Serum Samples ◽  
N Protein ◽  
Fluorescent Microsphere ◽  
Microsphere Immunoassay

ABSTRACTFor effective disease surveillance, rapid and sensitive assays are needed to detect antibodies developed in response to porcine reproductive and respiratory syndrome virus (PRRSV) infection. In this study, we developed a multiplexed fluorescent microsphere immunoassay (FMIA) for detection of PRRSV-specific antibodies in oral fluid and serum samples. Recombinant nucleocapsid protein (N) and nonstructural protein 7 (nsp7) from both PRRSV genotypes (type I and type II) were used as antigens and covalently coupled to Luminex fluorescent microspheres. Based on an evaluation of 488 oral fluid samples with known serostatus, the oral fluid-based FMIAs achieved >92% sensitivity and 91% specificity. For serum samples (n= 1,639), the FMIAs reached >98% sensitivity and 95% specificity. The assay was further employed to investigate the kinetics of the antibody response in infected pigs. In oral fluid, the N protein was more sensitive for the detection of early infection (7 and 14 days postinfection), but nsp7 detected a higher level and longer duration of antibody response (28 days postinfection). In serum, the antibodies specific to nsp7 and N proteins were detected as early as 7 days postinfection, and the responses lasted more than 202 days. This study provides a framework from which a more robust assay could be developed to profile the immune response to multiple PRRSV antigens in a single test. The development of oral fluid-based diagnostic tests will change the way we survey diseases in swine herds and improve our ability to cheaply and efficiently track PRRSV infections in both populations and individual animals.

scholarly journals Molecular detection of Porcine Reproductive and Respiratory Syndrome Virus, Porcine Circovirus 2 and Hepatitis E Virus in oral fluid compared to their detection in faeces and serum

2020 ◽  
Author(s):  
Jan Plut ◽  
Urska Jamnikar-Ciglenecki ◽  
Marina Stukelj
Keyword(s):  
Hepatitis E Virus ◽  
Pathogen Detection ◽  
Oral Fluid ◽  
Hepatitis E ◽  
Viral Disease ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus ◽  
Respiratory Syndrome Virus ◽  
Rt Pcr ◽  
Serum Samples

Abstract Background Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), Porcine Circovirus Type 2 (PCV2) and Hepatitis E virus (HEV) are common and economically important viral disease causative agents detected in pig oral fluid (OF), faeces and serum at some infection stages. The purpose of this study was to detect PRRSV, PCV2 and HEV on six pig farms to determine which of the three sample types, OF, faeces or serum is appropriate for the diagnosis of these viruses in different pig categories. The following pig categories were included: 5 weeks-old (w/o), 7 w/o, 9 w/o, 11 w/o weaners, fatteners and breeding sows. Pursuant to the preliminary detection of each pathogen at the selected farms, OF samples, faeces, serum pools and 10 individual sera were examined, using PCR, for each age category. If any of the viruses were found in pools of faeces and OF, then faeces and OF from positive farms were tested separately for each pig category. The viral nucleic acids were detected using RT-PCR, PCR and real-time RT-PCR, for PRRSV, PCV2 and HEV respectively.Results PRRSV and HEV were detected on one farm and PCV2 on three others, positive results being more often obtained from the OF than from the faeces of the same animals. Ten individual serum samples from pigs from the same group of animals were also tested. The viruses were detected in almost all individual sera and OF in the same pig category with some exceptions: PRRSV was detected in the OF of fatteners but was absent in their sera; on Farm 2, PCV2 was detected in sera of 11 w/o pigs and fatteners but absent in group samples of their OF and, vice versa, in case of 9 w/o animals; HEV was detected in the OF of the youngest, 5 w/o weaners and absent in sera of the same age group.Conclusions The primary finding of the study is that OF is a welfare-friendly, non-invasive and highly efficient matrix for pathogen detection, thus evidencing the usefulness of pig OF as a matrix in which each of the three viruses considered can be detected with the highest probability.

scholarly journals PRRSV detection by qPCR in processing fluids and serum samples collected in a positive stable breeding herd following mass vaccination of sows with a modified live vaccine

2021 ◽  
Vol 7 (1) ◽  
Author(s):  
A. Lebret ◽  
P. Berton ◽  
V. Normand ◽  
I. Messager ◽  
N. Robert ◽  
...  
Keyword(s):  
The United States ◽  
Live Vaccine ◽  
Mass Vaccination ◽  
Respiratory Syndrome Virus ◽  
Serum Samples ◽  
Blood Samples ◽  
Stability Monitoring ◽  
Breeding Herd ◽  
Modified Live Vaccine ◽  
Processing Fluid

AbstractIn the last two decades, in France, Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) stabilization protocols have been implemented using mass vaccination with a modified live vaccine (MLV), herd closure and biosecurity measures. Efficient surveillance for PRRSV is essential for generating evidence of absence of viral replication and transmission in pigs. The use of processing fluid (PF) was first described in 2018 in the United States and was demonstrated to provide a higher herd-level sensitivity compared with blood samples (BS) for PRRSV monitoring. In the meantime, data on vertical transmission of MLV viruses are rare even as it is a major concern. Therefore, veterinarians usually wait for several weeks after a sow mass vaccination before starting a stability monitoring. This clinical study was conducted in a PRRSV-stable commercial 1000-sow breed-to-wean farm. This farm suffered from a PRRS outbreak in January 2018. After implementing a stabilisation protocol, this farm was controlled as stable for more than 9 months before the beginning of the study. PF and BS at weaning were collected in four consecutive batches born after a booster sow mass MLV vaccination. We failed to detect PRRSV by qPCR on PF and BS collected in a positive-stable breeding herd after vaccination with ReproCyc® PRRS EU (Boehringer Ingelheim, Ingelheim, Germany).

scholarly journals Epidemiological investigation of porcine circovirus type 2 and its coinfection rate in Shandong province in China from 2015 to 2018

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Zicheng Ma ◽  
Mengda Liu ◽  
Zhaohu Liu ◽  
Fanliang Meng ◽  
Hongyu Wang ◽  
...  
Keyword(s):  
Pseudorabies Virus ◽  
Porcine Circovirus Type ◽  
Porcine Circovirus Type 2 ◽  
Shandong Province ◽  
Porcine Circovirus ◽  
Respiratory Syndrome Virus ◽  
Limited Information ◽  
Serum Samples ◽  
Diarrhea Virus

Abstract Background Porcine circovirus type 2 (PCV2) is one of the crucial swine viral pathogens, caused porcine circovirus associated diseases (PCVAD). Shandong province is one of the most important pork producing areas and bears a considerable economic loss due to PCVAD. However, there is limited information on epidemiology and coinfection rate of PCV2 with other critical swine diseases in this area, such as porcine reproductive and respiratory syndrome virus (PRRSV), classical swine fever virus (CSFV), Pseudorabies virus (PRV), and porcine epidemic diarrhea virus (PEDV). Results Overall, 89.59% serum samples and 36.98% tissue samples were positive for PCV2 specified ELISA and PCR positive for PCV2, respectively. The coinfection rates of PCV2 with PRRSV, PRV, CSFV, and PEDV were 26.73%, 18.37%, 13.06%, and 3.47%, respectively. Moreover, genetic characteristic of PCV2 were analyzed based on the cap genes showing that PCV2d is the dominant sub-genotype circulating in the province. Conclusions Our findings reveal that PCV2d, as the dominant strain, is prevailing in pig farms in Shandong province at high levels. There was a high frequency of coinfection of PCV2 and PRRSV.

Porcine reproductive and respiratory syndrome virus (PRRSV) in serum and oral fluid samples from individual boars: Will oral fluid replace serum for PRRSV surveillance?

2010 ◽  
Vol 154 (1-2) ◽  
pp. 170-176 ◽  
Author(s):  
Apisit Kittawornrat ◽  
John Prickett ◽  
Wayne Chittick ◽  
Chong Wang ◽  
Mark Engle ◽  
...  
Keyword(s):  
Oral Fluid ◽  
Respiratory Syndrome Virus

scholarly journals Lack of evidence of porcine reproductive and respiratory syndrome virus (PRRSV) infection in domestic swine in Brazil

2004 ◽  
Vol 34 (2) ◽  
pp. 449-455 ◽  
Author(s):  
Janice Reis Ciacci-Zanella ◽  
Cristiano Trombetta ◽  
Ildara Vargas ◽  
Denise Euclydes Mariano da Costa
Keyword(s):  
Genetic Material ◽  
Elisa Test ◽  
Respiratory Syndrome Virus ◽  
Rt Pcr ◽  
Serum Samples ◽  
Viral Isolation ◽  
Culture Cells ◽  
Commercial Kits ◽  
Swine Herds ◽  
Domestic Swine

This report describes the first prevalence of antibodies and experimental inoculation of suspected samples of porcine reproductive and respiratory syndrome virus (PRRSV) from ELISA positive pigs from swine herds in Brazil. Based on the hypothesis that this agent is present in swine herds worldwide, the objective of this work was to establish a diagnostic methodology and to investigate the occurrence of PRRSV in Brazilian swine herds. Fifty-four swine herds, the total number which imported genetic material (live pigs or swine semen) from countries where PRRS was endemic from 1990 to December 2000, from eight Brazilian States all included in this study. The sampling used was such as to detect a prevalence of infection of 5%, with a confidence level of 95%. A total of 3785 serum samples were tested for PRRSV antibodies by ELISA. Following the ELISA test, which was performed with two different commercial kits, all serum positive pigs were retested, examined and additional materials were collected. Viral isolation in permissive tissue culture cells and swine bioassays were performed. Additionally, reverse transcriptase polymerase chain reaction (RT-PCR) and nested RT-PCR were also performed. We could not demonstrate the presence of PRRSV or RNA of PRRSV by viral isolation or RT-PCR (or nested RT-PCR), respectively in all of the analyzed samples. Furthermore, the pigs inoculated with PRRSV suspicion samples did not seroconvert nor produce characteristic PRRS lesions in the swine bioassay. Thus, our results indicate no evidence of PRRSV in the samples analyzed from swine herds in this study.

Thyroid hormone suppression in feeder pigs following polymicrobial or porcine reproductive and respiratory syndrome virus-2 challenge

2021 ◽  
Author(s):  
J Alex Pasternak ◽  
Daniel J MacPhee ◽  
Joan K Lunney ◽  
Raymond R R Rowland ◽  
PigGen Canada ◽  
...  
Keyword(s):  
Viral Load ◽  
Thyroid Hormone ◽  
Thyroid Hormones ◽  
Average Daily Gain ◽  
Respiratory Syndrome Virus ◽  
Post Inoculation ◽  
Serum Samples ◽  
Cross Sectional ◽  
Post Exposure ◽  
Daily Gain

Abstract Thyroid hormones are powerful regulators of growth, development and basal metabolic rate and can be dysregulated under conditions of severe stress or illness. To understand the role of these hormones in porcine disease response, serum samples were obtained from 3 batches of nursery-aged pigs (n=208) exposed to a natural polymicrobial disease challenge with an array of bacterial and viral pathogens. Levels of total thyroxin (T4) and triiodothyronine (T3) assessed in sera by radioimmunoassay (RIA), decreased significantly by 14 days post exposure (DPE). Levels of T3 partially rebounded by 48 DPE, while T4 levels remain depressed. Post-exposure T3 and T4 levels were positively correlated with acute and long-term average daily gain. Cross-sectional sampling of animals maintained at the high health source farms, showed no equivalent change in either hormone when managed under standard industry conditions. To further elucidate the effect of PRRSV-infection on thyroid hormone levels, archived sera over 42 days post inoculation (DPI) from nursery pigs (N=190) challenged with one of two PRRSV2 strains by the PRRS Host Genetics Consortium (PHGC) were similarly assessed, with animals selected in a two-by-two design, to investigate biological extremes in average daily gain (ADG) and viral load. All animals showed a similar decrease in both thyroid hormones reaching a minimum at 7 DPI and returning to near pre challenge levels by 42 DPI. Post challenge T3 and T4 levels were significantly greater in high ADG groups, with no significant association with viral load or strain. The results of this study demonstrate porcine susceptibility to thyroid disruption in response to disease challenge and demonstrate a relationship between this response and growth performance.

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