Porcine Reproductive And Respiratory Syndrome Virus Antibody Research Articles

Quantum dot fluorescent microsphere-based immunochromatographic strip for detecting PRRSV antibodies

Porcine reproductive and respiratory syndrome (PRRS) is an immunosuppressive disease caused by the porcine reproductive and respiratory syndrome virus (PRRSV). Current vaccine prevention and treatment approaches for PRRS are not adequate, and commercial vaccines do not provide sufficient cross-immune protection. Therefore, establishing a precise, sensitive, simple, and rapid serological diagnostic approach for detecting PRRSV antibodies is crucial. The present study used quantum dot fluorescent microspheres (QDFM) as tracers, covalently linked to the PRRSV N protein, to develop an immunochromatography strip (ICS) for detecting PRRSV antibodies. Monoclonal antibodies against PRRSV nucleocapsid (N) and membrane (M) proteins were both coated on nitrocellulose membranes as control (C) and test (T) lines, respectively. QDFM ICS identified PRRSV antibodies under 10 min with high sensitivity and specificity. The specificity assay revealed no cross-reactivity with the other tested viruses. The sensitivity assay revealed that the minimum detection limit was 1.2 ng/mL when the maximum dilution was 1:2,048, comparable to the sensitivity of enzyme-linked immunosorbent assay (ELISA) kits. Moreover, compared to PRRSV ELISA antibody detection kits, the sensitivity, specificity, and accuracy of QDFM ICS after analyzing 189 clinical samples were 96.7%, 97.9%, and 97.4%, respectively. Notably, the test strips can be stored for up to 6 months at 4 °C and up to 4 months at room temperature (18–25 °C). In conclusion, QDFM ICS offers the advantages of rapid detection time, high specificity and sensitivity, and affordability, indicating its potential for on-site PRRS screening.Key points• QDFM ICS is a novel method for on-site and in-lab detection of PRRSV antibodies• Its sensitivity, specificity, and accuracy are on par with commercial ELISA kits• QDFM ICS rapidly identifies PRRSV, aiding the swine industry address the evolving virus

Reproductive features of PRRS-convalescent large white pigs after porcine reproductive and respiratory syndrome

Most sows convalescent from porcine reproductive and respiratory syndrome (PRRS) are immune to the re-infection. The PRRS virus's antibodies detected by enzyme-linked immunosorbent assay (ELISA) can persist for a year. The PRRS virus (PRRSV) causes damage to the pigs' reproductive system, manifested by abortions, delivery of non-viable piglets and saws' infertility. We carried out morphometric studies of recovered pregnant pigs' reproductive organs to determine changes in the fetus and placenta system in pregnant PRRS-convalescent sows. Morphological studies of the fetal portion of the placenta indicate that the weight of the placenta in PRRS-convalescent sows was significantly lower as compared to the weight of the placenta in clinically healthy pregnant sows, 0.71 ± 0.05 kg versus 0.92 ± 0.09 kg at the end of the second trimester, and 1.61 ± 0.42 versus 1.75 ± 0.16 kg on day 105-110 of gestation. By day 70-75 of gestation, we observed a significant 31.8%-decrease in the fetus's body weight in seropositive pigs compared to the clinically healthy ones and 19.2%-decrease on days 105-110. Piglets born from the experimental sows demonstrated physiological abnormalities that allowed for hypotrophy diagnosis. ELISA-tests of sera for anti-Müllerian hormone (AMH) proved the possibility of early assessment of gilts' fertility. This method will enable the premature culling of low-yielding replacement gilts from the breeding stock. Further development of the methods for gilts' fertility determination and ELISA-testing for AMH during the mandatory gynecological screening of the gilts at the pig breeding establishments are economically practical and effective.

Farm characteristics and sero-prevalence of porcine reproductive and respiratory syndrome virus (PRRSV) antibodies in pigs of Nepal.

Porcine reproductive and respiratory syndrome is a highly infectious disease of swine caused by PRRS virus (PRRSV). To evaluate the prevalence of PRRSV antibodies in the four districts of hilly and terai regions of Nepal. Toassess the farm characteristics through a questionnaire interview of farmersregarding management practices and PRRS. A cross-sectional study was conducted from July 2020 to June 2021 to determine the sero-prevalence of PRRSV in pigs. A total of 180 porcine serum samples were collected from 23 pig farms and tested for PRRSV antibodies by ELISA. Alongside, farm characteristics were also assessed through questionnaire to determine the level of biosecurity measures in the farm, knowledge of the disease and possible control mechanisms. Out of 180 samples, 37 were tested positive resulting the overall sero-prevalence of 20.5%. There was significant association between different districts (p < 0.05) and PRRS prevalence. Prevalence of PRRSV antibody was found higher in Kaski district (10.5%) followed by Sunsari (8.8%) district. Based on age groups, highest prevalence was found in age groups of above 18 months (9.4%), followed by 13-18 months age groups (7.7%). Regarding the knowledge level of the disease, 43% of the farmers responded that they have heard about the disease. Biosecurity practices in the farm was found very poor where only 40% of the farms had disinfectant at the entrance of the farm and 25% pig farmers were found using separate boots while dealing with pigs. The findings of this study reveal the presence of PRRSV antibodies in pigs of Nepal. In addition poor biosecurity measures, management practices and poor knowledge level about the disease among farmers highly affect in the control and prevention of disease thereby affecting the pig production and productivity. Therefore, government should develop and implement effective control measures and biosecurity programs.

Considerations in the use of processing fluids for the detection of PRRSV RNA and antibody.

Surveillance is mandatory for tracking the progress of porcine reproductive and respiratory syndrome virus (PRRSV) control and elimination efforts in breeding herds. Processing fluids, the fluid recovered from tissues collected at castration and/or tail docking, are used for breeding herd surveillance by large segments of the industry, but the basic diagnostic characteristics of processing fluids are largely undescribed. We undertook 3 studies to address this information gap. In study 1, we found no differences among the PRRSV RT-rtPCR results obtained with 4 commercial RNA extraction kits. In study 2, we found that PRRSV RNA was highly stable in processing fluid samples at -20°C or 4°C, but detrimental effects were observed at ≥22°C within 24 h. In study 3, using a modified PRRSV ELISA at a sample:positive cutoff of ≥0.5, we found excellent discrimination in the detection of PRRSV antibody (IgM, IgA, IgG) in processing fluids from herds of known PRRSV status. Judicious handling of processing fluid samples from sow herds, and the use of methods available in veterinary diagnostic laboratories, can provide a foundation for reliable PRRSV surveillance.

Risk Factors and Spatial-Temporal Analysis of Porcine Reproductive and Respiratory Syndrome Seroprevalence in China Before and After African Swine Fever Outbreak.

Porcine reproductive and respiratory syndrome (PRRS) is an infectious viral disease that causes great harm to the pig industry. PRRS virus (PRRSV), the causative agent of PRRS, is characterized by severe reproductive failure and respiratory confusion. This study performed a cross-sectional investigation of PRRSV seroprevalence and collected 14,134 serum samples in pig farms without PRRSV vaccination from 12 provinces and two cities in China from 2017 to 2021 to detect PRRSV antibodies by enzyme-linked immunosorbent assay (ELISA). The apparent and true PRRSV antibody prevalence was estimated and compared based on the Clopper-Pearson method and Pearson chi-square test, respectively. Risk factors associated with the PRRSV serological status of pig farms were analyzed through univariate and multivariable logistic regression analysis. An automatic autoregressive integrated moving average (ARIMA) model procedure was used for time-series analysis for PRRSV seroprevalence. Spatial clusters of high PRRSV seroprevalence were detected by SaTScan software. The total true PRRSV seroprevalence of the animal level was 62.56% (95% confidence interval [CI]: 61.74–63.37%). Additionally, 286 out of 316 pig farms were positive for PRRSV antibodies at the herd level. Pig farms without pseudorabies virus (PRV) infection were 5.413 (95% CI: 1.977–17.435) times more likely to be PRRSV antibody positive than those with PRV. Identically, the possibility of pig farms being PRRSV antibody positive before an African swine fever (ASF) outbreak was 3.104 (95% CI: 1.122–10.326) times more than after ASF. The odd ratio values of medium and large pig farms with PRRSV infection are 3.076 (95% CI: 1.005–9.498) and 6.098 (95% CI: 1.814–21.290). A fluctuant decline pattern for PRRSV prevalence was observed in the temporal analysis. Three significant clusters of high PRRSV seroprevalence were first detected in China, covering a time frame from January 2018 to September 2018, which reveals high PRRSV prevalence before the outbreak of ASF. These findings show the epidemic situation and spatial-temporal distribution of PRRSV infection in China in recent years and could help develop reasonable measures to prevent PRRSV infection.

Seroprevalence of antibodies to classical swine fever virus and porcine reproductive and respiratory syndrome virus in healthy pigs in Hunan Province, China.

Classical swine fever (CSF) and porcine reproductive and respiratory syndrome (PRRS) are responsible for major economic losses and represent a threat to the swine industry worldwide. Routine surveillance serology for CSF and PRRS viruses is critical to maintaining the health status of sow farms in Hunan Province, which is one of the top pig production provinces in China. The aim of our study was to investigate the serological statistics of CSF virus (CSFV) and PRRS virus (PRRSV) in Hunan Province. The cohort serum samples were collected from vaccinated and unvaccinated pigs. Our findings showed that the average rates of CSFV and PRRSV antibody seropositivity were 82.2% (95% CI: 80.1-84.3) and 84.8% (95% CI: 82.5-87.1), respectively, in the immunized group and that these rates were higher than those in the unvaccinated group (58.6% for CSFV and 47.8% for PRRSV). Additionally, the level of CSFV antibody in piglet serum declined gradually with age, whereas PRRSV-specific antibody level increased initially (1 to 2 weeks old) and then declined with age (2 to 4 weeks old). In summary, we investigated the difference in CSFV/PRRSV antibody levels among piglets at various weeks old (1 to 4 weeks) to further establish the duration of maternal immunity in piglets. In addition, routine monitoring of CSFV/PRRSV antibodies in immunized pigs was carried out to evaluate the efficacy of vaccination.

A serological survey of porcine reproductive and respiratory syndrome virus in wild boar in Gifu Prefecture, Japan.

Porcine reproductive and respiratory syndrome (PRRS) is an infectious swine disease caused by the PRRS virus (PRRSV) that results in economic loss to the pig-rearing industry. To study PRRSV infection in wild boars and pigs, we conducted a serological survey in Gifu Prefecture, Japan, from 2020 to 2021. Three out of 453 (0.7%) wild boar sera were positive for PRRSV antibodies in a commercial ELISA. However, given that PRRSV RNA was not detected in these three wild boars and the specificity and sensitivity of the test kit, these are considered as false positives. Although seropositive pigs were found in multiple pig farms in the study area, the role of wild boars as a source of PRRS to pig farms appeared to be minimal.

Novel DNA methylation markers of PRRSV-specific antibodies and their intergenerational transmission from pregnant sows to piglets.

The main strategy for preventing porcine reproductive and respiratory syndrome (PRRS) is vaccination. However, current commercial porcine reproductive and respiratory syndrome virus (PRRSV) vaccines have limited effectiveness and may even cause infections in pigs. The identification of stable molecular markers associated with immune responses to PRRSV vaccination in pigs provides a new approach for PRRS prevention. DNA methylation, the most stable epigenetic molecular marker related to PRRSV vaccination, has not been investigated. In the current research, we used whole genome bisulfite sequencing (WGBS) to investigate DNA methylation in pregnant sows that received PRRSV vaccination and their piglets with high and low PRRSV-specific antibody levels. By performing methylation data analysis and basing on our previous transcriptomic studies, we identified several differentially methylated genes (DMGs) that are involved in the pathways of inflammatory and immune responses. Among the DMGs, ISG15, MX1, SERPINE1, GNG11 and IFIT3 were common hub genes in the two generations. MX1 and GNG11 were located in quantitative trait loci related with PRRSV antibody titer and PRRSV susceptibility, respectively. These results suggest that PRRSV vaccination in sows induces DNA methylation changes in genes and DNA methylation changes occur through intergenerational transmission. The novel DNA methylation markers and target genes observed in our study provide new insights into the molecular mechanisms of immune responses to PRRSV vaccination across two pig generations.

A retrospective study of porcine reproductive and respiratory syndrome virus infection in Brazilian pigs from 2008 to 2020.

Porcine reproductive and respiratory syndrome (PRRS) is a viral disease characterized by reproductive impairment or failure in breeding animals, and a respiratory disease in pigs of any age. Brazil is the fourth largest pork producer and exporter globally, and PRRS virus (PRRSV) infection has never been reported in the country. This study aimed to investigate the status of porcine biological samples from commercial swine herds, quarantined imported boars, wild boars and feral pigs to update PRRS information in Brazil. A total of 14,382 samples were collected from 2008 to 2020, including sera (n=12,841), plasma (n=1,000) and oral fluids (n=541), comprehending 137 herds and free-living pigs in eight Brazilian states. One out of 1,000 (0.1%) plasma and 15 out of 12,841 (0.11%) serum samples tested positive for PRRSV antibodies through ELISA. Upon ELISA retesting, only the plasma sample, from one 8-day-old piglet remained positive. All sixteen previously PRRSV antibody-positive samples were tested through RT-PCR and found to be negative. The presence of false-positive or singleton reactors are quite expected. Thus, the use of different/alternative diagnostic tests is indicated for an efficient PRRSV detection. Taken together, our findings demonstrated no conclusive evidence of PRRSV infection in the tested pigs, highlighting the importance to reinforce the surveillance program to prevent the introduction and eventual dissemination of PRRSV in Brazil.

Association between management factors and herd status regarding Porcine Reproductive and Respiratory Syndrome virus infection - An analysis in the course of a voluntary PRRSV control program in Saxony and Thuringia

In Saxony and Thuringia, federal states of Germany with a low density of commercial pig farms, a voluntary program aims at controlling porcine reproductive and respiratory syndrome virus (PRRSV) infection. This targets the eradication of the infection on a herd level which has previously been achieved in a subset of herds. The presented study aimed at identifying management factors related with a positive or a negative PRRSV antibody (AB status) or PRRSV genome status (PCR status) on a herd level. Data were collected in 82 farms in a region implementing a voluntary PRRSV control program. The test findings for the years 2011 to 2018 were compiled for each year and associated with the interrogated parameters. A generalized linear mixed model was used to identify factors associated with the AB and PCR status. The variables "separation of contaminated and non-contaminated areas on the loading ramp" (p = 0.012), "separation of gilts and sows" (p = 0.017) and "recording of visitors in a book" (p = 0.046) were negatively associated with the PCR status. In contrast, "separation of gilts and finishers" (p = 0.044) as well as the existence of "separated alleyways" (p = 0.042) were positively related to the PCR status. "Vaccination against PRRSV" was positively associated with the AB status and the PCR status (p = 0.005 and p = 0.001, respectively). In numerous variables, a low variability was observed. Certain biosecurity measures to control the movement of animals (separation of contaminated and not contaminated areas on the loading ramp) or people (recording of visitors) contribute to a successful reduction of PRRSV infections and a negative herd status. A combination of different measures may reduce PRRSV spread within pig herds. Breaking the infection cycle in gilts, either by separation of gilts from older sows or immunization, may be considered as a key aspect, presumably additionally supported by keeping gilts together with fattening pigs.

Evidence of transboundary transmission of viral pathogens through pigs transported from Myanmar to Mizoram, India

In Mizoram, pigs used to migrate through the porous international border from Myanmar. It is speculated that the migrated animals may act as potential carriers of various microbial pathogens. But so far no scientific evidence is available in this aspect. The present study was conducted on the detection of serum antibodies and molecular detection of classical swine fever virus (CSFV), porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus 2 (PCV2) and rotavirus in the pigs. A total of 204 blood and 209 rectal swabs were collected from apparently healthy pigs of Champhai district of Mizoram transported from Myanmar. All the serum samples were subjected to ELISA based detection of antibodies against CSFV, PRRSV and PCV2. The whole blood/serum/feces were processed for extraction of viral nucleic acids and subjected to PCR/RT-PCR for detection of CSFV, PRRSV, PCV2 and rotavirus. A total of 105 (51.47%), 12(5.88%) and 36 (17.65%) serum samples were found to be positive for antibodies against CSFV, PRRSV and PCV2, respectively. In addition, 17 (8.33%) sera were recorded positive for both CSFV and PCV-2 antibodies and 9 (4.41%) sera were positive for CSFV, PCV-2 and PRRSV antibodies. By specific PCR, group A rotavirus was found to be most prevalent (8.33%) followed by PCV-2 (7.84%), CSFV (4.41%) and PRRSV (0.9%). All the rotaviruses were detected as Group A only. Of the 204 animals, 2 (0.98%) pigs showed co-infection with all the four viruses under the study. A total of 5 (2.45%) pigs were positive for co-infection with CSFV and PCV-2 and 4 (1.96%) pigs were positive for co-infection with PCV-2 and rotavirus. The present study provided laboratory based evidence on transmission of porcine viral pathogens from Myanmar to Mizoram.

Porcine reproductive and respiratory syndrome virus (PRRSV) antibody levels in large swine farms in selected regions of Yunnan province, China

Porcine reproductive and respiratory syndrome (PRRS) is caused by the PRRS virus (PRRSV), and it is a widespread disease that severely affects swine production in all age groups. Detection of PRRSV antibody levels in pig farms is beneficial for immunity evaluations. In this study, a total of 1,206 serum samples of breeding boars, breeding sows, reserve pigs and commercial pigs from 16 large-scale swine farms in 4 different regions of Yunnan province in China were collected during 2019 and detected by indirect enzyme-linked immunosorbent assay (ELISA). The results showed that the average positive rate of PRRSV antibody was 88.32%, among which the antibody-positive rates were 89.03%, 89.18%, 92.11%, and 82.95% in East Yunnan (E. Yunnan), Central Yunnan (Cent. Yunnan), Northwest Yunnan (N.W. Yunnan) and Northeast Yunnan (N.E. Yunnan), respectively (P &gt; 0.05). For the different pig categories, the reserve pigs (93.51%) showed much higher antibody-positive rates, followed by breeding sows (92.44%), commercial pigs (87.34%) and breeding boars (85.62%). Statistical analysis revealed that the rates were significantly different among different pig categories (P &lt;0.05). These s results indicated that pig categories were significantly associated with PRRSV antibody levels in this study. All the positive rates in this study fulfilled the requirement of ≥ 70% set by the National Animal Disease Surveillance Plan of China (2011). The study could provide evidence of the antibody response of PRRSV at the farm level.

Porcine reproductive and respiratory syndrome control in Saxony and Thuringia - Results of 2 voluntary regional programmes in the years 2011-2018

The present study is intended to show the progress of voluntary porcine reproductive and respiratory syndrome (PRRS) control programmes in Thuringia and Saxony as well as to emphasize the importance of confirmation testing in the case of positive findings in unsuspicious herds. In order to present the progress of porcine reproductive and respiratory syndrome virus (PRRSV) elimination, the findings for the years 2011-2018 as well as data on the elimination programme, type and size of the farms, and use of PRRS vaccination were compiled. Each herd was assigned an antibody and PCR status for each year in order to analyze the relationship between status and year. Furthermore, the association between status and herd size as well as the use of PRRS vaccination were investigated. The relevance of confirmation testing is shown by the findings of the herds in the years following PRRSV elimination. In the majority of affected farms in Saxony, depopulation/repopulation was performed in order to eliminate PRRSV. In Thuringia, PRRSV elimination was frequently accomplished during ongoing production. A significant reduction of positive antibody and PCR findings was observed in both federal states during the investigation period. Higher antibody and PCR levels were evident in farms using PRRS vaccination. In the years following PRRSV elimination, positive PRRSV antibody findings occurred in individual PRRSV-unsuspicious farms. These were shown to be false-positive by use of confirmation tests. The proportion of PRRSV-positive herds in a region with low pig density can be reduced by voluntary control programmes with the eradication method playing a subordinate role. The use of vaccination against PRRSV or alternative control measures is to be considered carefully. In the case of positive findings in unsuspicious farms, confirmation testing is warranted in order to rule out any ongoing PRRS outbreak as well as to clarify the farm's status. Voluntary PRRS elimination and maintenance of the status of "PRRSV unsuspicious" is possible even when individual herds in the region do not participate in the control programme.

Development of recombinant nucleocapsid protein-based indirect enzyme-linked immunosorbent assay for sero-survey of porcine reproductive and respiratory syndrome.

Background and Aim:Porcine reproductive and respiratory syndrome (PRRS) is a disease endemic in many countries and is of economic importance. India was free from PRRS until the first outbreak was reported from a North-East Indian state in 2013. Since then, disease outbreaks have been reported from North-East India and the pilot study conducted earlier showed that it is gradually spreading to the rest of India. Considering there are no locally developed population screening tests available for PRRS and imported diagnostic/screening tests are expensive, the present study was aimed at developing recombinant nucleocapsid (rN) protein-based indirect enzyme-linked immunosorbent assay (iELISA).Materials and Methods:The rN protein of PRRS virus (PRRSV) was produced following standard cloning, expression, and purification procedures. Using this antigen, iELISA was optimized for the detection of serum antibodies to PRRSV. The sensitivity and specificity of the test were assessed by comparing it with a commercial PRRSV antibody detection kit.Result:A total of 745 serum samples from ten different states of India were screened using the developed iELISA. The iELISA had a relative specificity of 76.18% and sensitivity of 82.61% compared to the commercial ELISA (Priocheck PRRSV ELISA kit, Thermo Fisher Scientific, USA).Conclusion:The iELISA, which deployed rN protein from Indian PRRSV, was found to be suitable in the serological survey and may be a useful tool in future disease surveillance programs.

A rapid and quantitative fluorescent microsphere immunochromatographic strip test for detection of antibodies to porcine reproductive and respiratory syndrome virus.

A fluorescent microsphere-based immunochromatographic strip test (FICT) was developed for the rapid, sensitive, and quantitative detection of porcine reproductive and respiratory syndrome virus (PRRSV) antibodies at the pen-side. The assay was based on the formation of a sandwich immune-complex (anti-pig IgG-PRRSV antibodies-NSP7/N), which was validated by a comparison with IDEXX-ELISA using 3325 clinical specimens. The diagnostic specificity, sensitivity, and accuracy of FICT were 97.28, 93.41, and 94.95%, respectively. FICT showed a good correlation with the virus neutralization assay. Overall, a promising pen-side diagnostic tool was developed for the rapid and quantitative detection of PRRSV antibodies within 15 min.

UV-C irradiation is able to inactivate pathogens found in commercially collected porcine plasma as demonstrated by swine bioassay

Liquid porcine plasma is an animal origin raw material for the manufacturing process of spray-dried porcine plasma that is used in pig nutrition worldwide. In previous studies we found that the application of ultraviolet light C (UV-C) in liquid plasma that was inoculated with a variety of bacteria or viruses of importance in the swine industry can be considered as redundant safety steps because in general achieve around 4 logs reduction for most of these pathogens. However, the final validation of the UV-C light as safety feature should be conducted with commercial liquid plasma and using the pig bioassay model. As a first objective, the potential infectivity of a raw liquid plasma product collected from an abattoir was tested by means of a swine bioassay. We used Porcine circovirus 2 (PCV-2), a ubiquitous virus that has been systematically detected by PCR in porcine plasma at abattoirs as selection criteria for commercial liquid plasma lot. As a second aim of the study, the effects of different doses of UV-C irradiation on the selected raw liquid plasma were assayed in the animal bioassay. Moreover, other swine infecting agents, including Porcine reproductive and respiratory syndrome virus (PRRSV), were also determined in the original plasma and monitored in the inoculated animals. Pigs negative for PCV-2 and PRRSV genome and antibodies were allotted to one of five groups (6 to 8 pigs/ group) and injected intra-peritoneally with 10 mL of their assigned inoculum at 50 d of age. Negative control pigs (group 1) were injected with PBS. Positive control pigs (group 5) were injected with a PCV-2 inoculum. Groups 2, 3 and 4 were injected with liquid porcine plasma that had been subjected to 0 (raw plasma), 3000 or 9000 J/L UV-C irradiation, respectively. Group 2 pigs (0 J/L UV-C) got infection by PRRSV but no PCV-2 infection or seroconversion. However, one pig from group 2 seroconverted to Rotavirus A (RVA) and Hepatitis E virus (HEV) and three group 2 pigs seroconverted to Porcine parvovirus (PPV). Groups 1, 3 and 4 pigs showed no evidence of infection or seroconversion associated with the tested viruses or any other pathogens found in the liquid plasma before UV-C irradiation. Group 5 pigs developed PCV-2 infectivity as expected. UV-C irradiation of liquid plasma at 3000 and 9000 J/L was effective in preventing PRRSV and other pathogens transmission. Moreover, raw liquid plasma was non-infectious for PCV-2 in naïve pigs.

Natural viral co-infections in pig herds affect hepatitis E virus (HEV) infection dynamics and increase the risk of contaminated livers at slaughter.

Hepatitis E virus (HEV) is a zoonotic pathogen, in particular genotype 3 HEV is mainly transmitted to humans through the consumption of contaminated pork products. This study aimed at describing HEV infection patterns in pig farms and at assessing the impact of immunomodulating co-infections namely Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) and Porcine Circovirus Type 2 (PCV2), as well as other individual factors such as piglets' immunity and litters' characteristics on HEV dynamics. A longitudinal follow-up was conducted in three farrow-to-finish farms known to be HEV infected. Overall, 360 piglets were individually monitored from birth to slaughter with regular blood and faecal sampling as well as blood and liver samples collected at slaughterhouse. Virological and serological analyses were performed to detect HEV, PCV2 and PRRSV genome and antibodies. The links between 12 explanatory variables and four outcomes describing HEV dynamics were assessed using cox-proportional hazard models and logistic regression. HEV infection dynamics was found highly variable between farms and in a lower magnitude between batches. HEV positive livers were more likely related to short time-intervals between HEV infection and slaughter time (<40days, OR=4.1 [3.7-4.5]). In addition to an influence of piglets' sex and sows' parity, the sequence of co-infections was strongly associated with different HEV dynamics: a PRRSV or PCV2/PRRSV pre- or co-infection was associated with a higher age at HEV shedding (Hazard Ratio=0.3 [0.2-0.5]), as well as a higher age at HEV seroconversion (HR=0.5 [0.3-0.9] and HR=0.4 [0.2-0.7] respectively). A PCV2/PRRSV pre- or co-infection was associated with a longer duration of shedding (HR=0.5 [0.3-0.8]). Consequently, a PRRSV or PCV2/PRRSV pre- or co-infection was strongly associated with a higher risk of having positive livers at slaughter (OR=4.1 [1.9-8.9] and OR=6.5 [3.2-13.2] respectively). In conclusion, co-infections with immunomodulating viruses were found to affect HEV dynamics in the farrow-to-finish pig farms that were followed in this study.

Comparison of six commercial ELISAs for the detection of antibodies against porcine reproductive and respiratory syndrome virus (PRRSV) in field serum samples

Porcine reproductive and respiratory syndrome (PRRS) is one of the most common infectious diseases of swine globally. Since the course of PRRS virus (PRRSV) infection is subclinical, laboratory diagnosis is necessary to detect the virus or specific antibodies. The aim of this study was to assess the sensitivity and specificity of IDEXX PRRS X3 Ab Test (IDEXX, USA), Civtest Suis E/S (Hipra, Spain), INgezim PRRS 2.0 (Ingenasa, Spain), VetExpert PRRS Ab ELISA 4.0 (BioNote, Korea), Pigtype PRRSV Ab (Qiagen, Germany) and PrioCHECK PRRSV Antibody ELISA (ThermoFisher, USA), using serum samples obtained from 5 conventional PRRSV-positive and 5 PRRSV-negative Polish pig farms.Specificity of ELISAs ranged from 94.2% (ThermoFisher) to 100% (IDEXX and Hipra). ThermoFisher ELISA had the highest detection rate and detected 67.2% samples from PRRSV-positive farms as positive but considering its low specificity some of the positive results may be incorrect. IDEXX ELISA considered as a reference detected 64.8% positive sera in PRRSV-positive farms. On the other hand Hipra Elisa identified only 51.8% of samples as positive. The diagnostic sensitivity of five ELISAs relative to IDEXX ranged from 80.3% (Hipra) to 96.3% (ThermoFisher).Our study showed significant differences in specificity and diagnostic sensitivity between the compared kits. The differences in the performance appeared to be practically negligible on farms where early infection with PRRSV occurred. However, on PRRSV-negative farms, or farms with PRRSV stable sow herds, some ELISAs can give results not reflecting the infection status in specific age groups.

Assessment of abattoir based monitoring of PRRSV using oral fluids

Various porcine reproductive and respiratory syndrome virus (PRRSV) regional elimination projects have been implemented in the U.S., but none have yet succeeded. In part, this reflects the need for efficient methods to monitor over time the progress of PRRSV status of participating herds. This study assessed the feasibility of monitoring PRRSV using oral fluids collected at the abattoir. A total of 36 pig lots were included in the study. On-farm oral fluid (n = 10) and serum (n = 10) collected within two days of shipment to the abattoir were used to establish the reference PRRSV status of the population. Oral fluids (n = 3 per lot) were successfully collected from 32 lots (89%) at the lairage. Three veterinary diagnostic laboratories (VDLs) tested the sera (VDL1 and VDL3: n = 316, VDL2: n = 315) and oral fluids (VDL1 and VDL3: n = 319, VDL2: n = 320) for PRRSV antibodies (ELISA) and RNA (rRT-PCR). Environmental samples (n = 64, 32 before and 32 after pigs were placed in lairage) were tested for PRRSV RNA at one VDL. All oral fluids (farm and abattoir) tested positive for PRRSV antibody at all VDLs. PRRSV positivity frequency on serum ranged from 92.4% to 94.6% among VDLs, with an overall agreement of 97.6%. RNA was detected on 1.3% to 1.9%, 8.1% to 17.7%, and 8.3% to 17.7% of sera, on-farm and abattoir oral fluids, respectively. Between-VDLs rRT-PCR agreement on sera and oral fluids (farm and abattoir) ranged from 97.8% to 99.0%, and 79.0% to 81.2%, respectively. Between-locations agreement of oral fluids varied from 31.3% to 50% depending on the VDL. This study reported the application of swine oral fluids collected at the abattoir for monitoring PRRSV, and describes the between-VDL agreement for PRRS testing of serum and oral fluid field samples.

Effect of chemical clarification of oral fluids on the detection of porcine reproductive and respiratory syndrome virus IgG.

Routine collection and testing of oral fluid (OF) samples facilitates porcine reproductive and respiratory syndrome virus (PRRSV) surveillance in commercial swine herds in a cost-effective, welfare-friendly fashion. However, OFs often contain environmental contaminants that may affect liquid handling and test performance. Traditional processing methods (e.g., filtration or centrifugation) are not compatible with high-throughput testing because of the burden of additional processing costs and time. OF "clarification" using chemical flocculants is an alternative approach not widely explored. Therefore, we evaluated the effect of chitosan-based clarification treatment on a commercial PRRSV OF ELISA. Serum and individual OFs were collected from vaccinated pigs ( n = 17) at -7 to 42 d post-vaccination and subdivided into 4 aliquots. Each aliquot was clarified (treatment A, B, C), with the 4th aliquot serving as untreated control. All samples were tested by PRRSV OF ELISA immediately after treatment and then were held at 4°C to be re-tested at 2, 4, 6, and 14 d post-treatment. Quantitative and qualitative treatment effects were evaluated. A Kruskal-Wallis test found no significant difference in ELISA S/P responses among treatments by days post-treatment. No difference was detected in the proportion of positive PRRSV antibody samples among treatments (Cochran Q, p > 0.05). Treatment of swine OFs using chitosan-based formulations did not affect the performance of a commercial PRRSV OF ELISA. Chitosan (or other flocculants) could improve OF characteristics and could be adapted for use in the field or in high-throughput laboratories.