Modified-live Porcine Reproductive And Respiratory Syndrome Virus Research Articles

Transition from one commercial porcine reproductive and respiratory syndrome modified-live virus vaccine to another in a breeding herd and impact on productivity

Porcine reproductive and respiratory syndrome (PRRS) continues to represent a significant cost to the swine industry and efforts are focused on prevention and mitigation of losses across production phases. Herein describes a PRRS modified-live virus (MLV) vaccinated breeding herd that changed commercial MLV vaccines to improve post-weaning performance. Two whole-herd vaccinations with a new PRRS MLV vaccine, administered 4 weeks apart, occurred without breeding herd production disruptions and with limited changes in diagnostic results. Replacement gilts tested PRRS virus negative 10 weeks post vaccination with the new MLV vaccine. Diagnostics were intermittently positive in the breeding herd and early nursery.

Histopathological Pulmonary Lesions in 1st-Day Newborn Piglets Derived from PRRSV-1 MLV Vaccinated Sows at the Last Stage of Gestation.

Modified live virus (MLV) vaccines for the control of porcine respiratory and reproductive syndrome virus (PRRSV) have been associated with the vertical and horizontal transmission of vaccine viruses. The present study aimed to describe pathological lung lesions in piglets born by gilts vaccinated with PRRSV-1 MLV. In total, 25 gilts were vaccinated at late gestation (100th day) and were divided into five groups according to the different vaccines (Vac) used: no vaccine-control group, Vac-1-strain DV, Vac-2-strain VP-046 BIS, Vac-3-strain 94881, Vac-4-strain 96V198. Within the first 0-9 h of the farrowing, blood samples were collected from all newborn piglets and lung samples were exanimated grossly, histopathologically and with scanning electron microscopy. PRRSV (RT-PCR-positive) and antibodies were detected in the serum of piglets from gilts vaccinated with Vac-2. In these piglets, moderate to severe interstitial pneumonia with thickened alveolar septa was noticed. Type II pneumocyte hyperplasia was also observed. The rest of the trial piglets showed unremarkable lung lesions. Phylogenetic analysis revealed the 98.7% similarity of the PRRSV field strain (GR 2019-1) to the PRRS MLV vaccine strain VP-046 BIS. In conclusion, the Vac-2 PRRSV vaccine strain can act as an infectious strain when vaccination is administrated at late gestation, causing lung lesions.

Comparison of the cross-protection of PPRSV sublineage 8.7 MLV vaccines against the recombinant NADC30-like strain

The emergence of recombinant porcine reproductive and respiratory syndrome virus (PRRSV) has caused a substantial threat to the swine industry in recent years. However, the protective efficacy of different sublineage 8.7 PRRSV modified-live virus (MLV) vaccines against emerging strains were still obscure. In this study, a broad epidemiological investigation of PRRSV showed the prevalence of NADC30-like strain increased in Shandong Province, China from 2018 to 2020. Through piglet trial for vaccination and challenge with recombinant NADC30-like SDlz1601 strain, CH-1R MLV vaccine showed better protective effect than JXA1-R and TJM-F92 MLV vaccines in terms of clinical score and pathological observation. Moreover, all three MLV vaccines could reduce virus loads in the serum of piglets. This study provides valuable insights into the prevalence of the NADC30-like strain and the protective effect of PRRS MLV vaccines against recombinant NADC30-like strains, which could help to improve the prevention and control of PRRSV infections.

Protection provided by a commercial modified-live porcine reproductive and respiratory syndrome virus (PRRSV) 1 vaccine (PRRSV1-MLV) against a Japanese PRRSV2 field strain.

Porcine reproductive and respiratory syndrome virus (PRRSV) vaccines do not provide full cross-protection, mainly due to the virus genetic variability. Despite this, vaccines based on modified-live PRRSV (PRRSV-MLV) reduce the disease impact. To assess the efficacy of two commercial vaccines-one based on PRRSV1 (PRRSV1-MLV) and another on PRRSV2 (PRRSV2-MLV)-against a Japanese PRRSV2 field strain. Two groups of three-week-old piglets were vaccinated (G1: PRRSV1-MLV; G2: PRRSV2-MLV) and two were kept as non-vaccinated (INF and CTRL). One month later, G1, G2, and INF were challenged with a PRRSV2 field strain. After the challenge, clinical signs were only observed in INF. Moreover, the highest rectal temperatures and values for the area under the curve (AUC) were observed in INF. Regarding viral detection, both AUC and the proportion of positive samples in blood were higher in INF. In G1, viremic animals never reached 100%. At necropsy (21 d after the challenge), differences for titers among groups were only found in tonsils (G1 < G2 and INF). One animal (belonging to G1) was negative in all tissues. Regarding humoral responses, G1 and G2 seroconverted after vaccination, as detected in the corresponding enzyme-linked immunosorbent assay. Specific neutralizing antibodies (NA) against PRRSV1-MLV were already detected at 14 d after vaccination in G1, showing a significant booster after the challenge, while PRRSV2-MLV NA were detected in G2 at the end of the experiment. Despite genetic differences, PRRSV1-MLV has been demonstrated to confer partial protection against a Japanese PRRSV2 strain, at least as good as PRRSV2-MLV.

Heterologous vaccine immunogenicity, efficacy, and immune correlates of protection of a modified-live virus porcine reproductive and respiratory syndrome virus vaccine.

Although porcine reproductive and respiratory syndrome virus (PRRSV) vaccines have been available in North America for almost 30 years, many vaccines face a significant hurdle: they must provide cross-protection against the highly diverse PRRSV strains. This cross-protection, or heterologous vaccine efficacy, relies greatly on the vaccine’s ability to induce a strong immune response against various strains—heterologous immunogenicity. Thus, this study investigated vaccine efficacy and immunogenicity of a modified live virus (MLV) against four heterologous type 2 PRRSV (PRRSV-2) strains. In this study, 60 pigs were divided into 10 groups. Half were MOCK-vaccinated, and the other half vaccinated with the Prevacent® PRRS MLV vaccine. Four weeks after vaccination, groups were challenged with either MOCK, or four PRRSV-2 strains from three different lineages—NC174 or NADC30 (both lineage 1), VR2332 (lineage 5), or NADC20 (lineage 8). Pre-and post-challenge, lung pathology, viral loads in both nasal swabs and sera, anti-PRRSV IgA/G, neutralizing antibodies, and the PRRSV-2 strain-specific T-cell response were evaluated. At necropsy, the lung samples were collected to assess viral loads, macroscopical and histopathological findings, and IgA levels in bronchoalveolar lavage. Lung lesions were only induced by NC174, NADC20, and NADC30; within these, vaccination resulted in lower gross and microscopic lung lesion scores of the NADC20 and NADC30 strains. All pigs became viremic and vaccinated pigs had decreased viremia upon challenge with NADC20, NADC30, and VR2332. Regarding vaccine immunogenicity, vaccination induced a strong systemic IgG response and boosted the post-challenge serum IgG levels for all strains. Furthermore, vaccination increased the number of animals with neutralizing antibodies against three of the four challenge strains—NADC20, NADC30, and VR2332. The heterologous T-cell response was also improved by vaccination: Not only did vaccination increase the induction of heterologous effector/memory CD4 T cells, but it also improved the heterologous CD4 and CD8 proliferative and/or IFN-γ response against all strains. Importantly, correlation analyses revealed that the (non-PRRSV strain-specific) serum IgG levels and the PRRSV strain-specific CD4 T-cell response were the best immune correlates of protection. Overall, the Prevacent elicited various degrees of efficacy and immunogenicity against four heterologous and phylogenetically distant strains of PRRSV-2.

Recombination between the Fostera MLV-like Strain and the Strain Belonging to Lineage 1 of Porcine Reproductive and Respiratory Syndrome Virus in Korea.

Porcine reproductive and respiratory syndrome virus (PRRSV) is one of the most important pathogens in the swine industry worldwide. In Korea, Fostera PRRS commercial modified live virus (MLV) vaccines have been used since 2014 to control the PRRSV infection. In this study, two PRRSV-2 strains (20D160-1 and 21R2-63-1) were successfully isolated, and their complete genomic sequences were determined. Genetic analysis showed that the two isolates have recombination events between the P129-like strain derived from the Fostera PRRS MLV vaccine and the strain of lineage 1. The 20D160-1 indicated that partial ORF2 to partial ORF4 of the minor parental KNU-1902-like strain, which belongs to Korean lineage C (Kor C) of lineage 1, was inserted into the major parental P129-like strain. The 21R2-63-1 revealed that partial ORF1b of the P129-like strain was inserted into the backbone of the NADC30-like strain. This study is the first to report natural recombinant strains between Fostera PRRS MLV-like strain and the field strain in Korea. These results may have significant implications for MLV evolution and the understanding of PRRSV genetic diversity, while highlighting the need for continuous surveillance of PRRSV.

Comparison of virus detection, productivity, and economic performance between lots of growing pigs vaccinated with two doses or one dose of PRRS MLV vaccine, under field conditions

Porcine reproductive and respiratory syndrome virus (PRRSV) causes a significant economic impact on swine production. It has been demonstrated that PRRS modified-live virus (MLV) vaccination of pigs, with one full dose, significantly reduces clinical consequences of wild-type PRRSV infection compared to non-vaccinates. However, there is limited information about the effect that two doses of PRRSV MLV vaccine have on the performance of growing pigs, compared to vaccination with a single dose. This study was conducted with the objectives to compare (a) the wild-type PRRSV detection in oral fluids over time, (b) key closeout productivity indicators, and (c) economic performance between lots of growing pigs vaccinated with two doses of Ingelvac PRRS® MLV vaccine and lots vaccinated with a single dose of the same vaccine. This randomized field trial included 15 lots of growing pigs from PRRSV positive-unstable sow farms and 66 lots from PRRSV positive-stable sow farms, according to the American association of swine veterinarians’ terminology. All pig lots received the first vaccination either around processing or weaning age. Lots allocated in the two doses group received the second vaccination three to four weeks after the first vaccination. The pig lots were monitored for PRRSV detection over time. Six oral fluids samples were collected in three weeks intervals and were tested for wild-type PRRSV-2 RNA by RT-qPCR and open reading frame 5 (ORF)− 5 sequencing. Regression models were used to compare wild-type PRRSV detection dynamics on oral fluids samples and to compare key closeout performance indicators between one dose group and two doses group. Additionally, a benefit-cost ratio analysis compared economic performance between one dose group and two doses group. The proportion of wild-type PRRSV detection on oral fluids samples and the log counts of viral RNA per ml of oral fluids from the two doses group was lower than the one dose group on lots originated from PRRSV positive-stable sow farms, with a risk ratio of 1.24 and a rate ratio of 1.17, respectively. The two doses group had a significantly lower mortality rate than the one dose group, with a rate ratio of 1.21. The effect size increased on lots originated from PRRSV positive-unstable sow farms, and on lots with higher frequency and diversity of wild-type PRRSV detection during the growth phase. No differences in growth performance were detected between two doses group and one dose group. The second MLV vaccination dose had a benefit-cost ratio of 1.83. For lots originated from PRRSV positive-unstable farms, the benefit-cost ratio was 4.45, and for lots originated from PRRSV positive-stable farms, the benefit-cost ratio was 0.45. Under study conditions, vaccinating growing pig lots with two doses of PRRS MLV vaccine was a useful strategy to immunize growing pigs against PRRSV, lowering the wild-type PRRSV detection, lowering mortality rate, and increasing profitability, compared to lots of growing pigs that received a single dose of the same vaccine.

Efficacy of the Synergy Between Live-Attenuated and Inactivated PRRSV Vaccines Against a NADC30-Like Strain of Porcine Reproductive and Respiratory Syndrome Virus in 4-Week Piglets.

The NADC30-like strain of porcine reproductive and respiratory syndrome virus (PRRSV) is a novel strain responsible for substantial economic losses to swine production in China. This study evaluated the cross-protective efficacy of the synergy between live-attenuated and inactivated PRRSV vaccines compared with a single vaccination with PRRS modified-live virus (MLV) vaccine against challenge with NADC30-like strain, v2016/ZJ/09-03. A total of 45 PRRSV free pigs were randomly divided into five groups: (1) strict control (SC); (2) positive control (PC); (3) single MLV dose (M1); (4) primed intramuscularly with MLV and boosted with killed vaccine 3 weeks later (MK1); and (5) intramuscular prime MLV boosted subcutaneously with killed vaccine B 3 weeks later (MK2). Serological tests in MK groups revealed no differences in both anti-N and anti-GP protein antibodies compared with M1 group, and failed to provide further protection against clinical signs, virus shedding, and gross lesions. However, the viremic titer, gross lung lesions, and average daily weight gain were significantly improved in the MLV vaccinated groups, suggesting that MLV provides substantial cross-protection against the NADC30-like virus. Thus, as a booster, the killed vaccine confers minimal additional protection in NADC30-like infected piglets.

Simultaneous Infection With Porcine Reproductive and Respiratory Syndrome and Influenza Viruses Abrogates Clinical Protection Induced by Live Attenuated Porcine Reproductive and Respiratory Syndrome Vaccination.

The porcine respiratory disease complex (PRDC) is responsible for significant economic losses in the pig industry worldwide. Porcine reproductive and respiratory syndrome virus (PRRSV) and swine influenza virus are major viral contributors to PRDC. Vaccines are cost-effective measures for controlling PRRS, however, their efficacy in the context of co-infections has been poorly investigated. In this study, we aimed to determine the effect of PRRSV-2 and swine influenza H3N2 virus co-infection on the efficacy of PRRSV modified live virus (MLV) vaccination, which is widely used in the field. Following simultaneous challenge with contemporary PRRSV-2 and H3N2 field isolates, we found that the protective effect of PRRS MLV vaccination on clinical disease and pathology was abrogated, although viral load was unaffected and antibody responses were enhanced. In contrast, co-infection in non-immunized animals reduced PRRSV-2 viremia and H3N2 virus load in the upper respiratory tract and potentiated T cell responses against both PRRSV-2 and H3N2 in the lung. Further analysis suggested that an upregulation of inhibitory cytokines gene expression in the lungs of vaccinated pigs may have influenced responses to H3N2 and PRRSV-2. These findings provide important insights into the effect of viral co-infections on PRRS vaccine efficacy that may help identify more effective vaccination strategies against PRDC in the field.

Gut microbiome associations with outcome following co-infection with porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) in pigs immunized with a PRRS modified live virus vaccine

Porcine reproductive and respiratory syndrome virus (PRRSV) and porcine circovirus type 2 (PCV2) are two of the most significant pathogens affecting swine. Co-infections are common and result in respiratory disease and reduced weight gain in growing pigs. Although PRRS modified live virus (MLV) vaccines are widely used to decrease PRRS-associated losses, they are generally considered inadequate for disease control. The gut microbiome provides an alternative strategy to enhance vaccine efficacy and improve PRRS control. The objective of this study was to identify gut microbiome characteristics associated with improved outcome in pigs immunized with a PRRS MLV and co-challenged with PRRSV and PCV2b. Twenty-eight days after vaccination and prior to co-challenge, fecal samples were collected from an experimental population of 50 nursery pigs. At 42 days post-challenge, 20 pigs were retrospectively identified as having high or low growth outcomes during the post-challenge period. Gut microbiomes of the two outcome groups were compared using the Lawrence Livermore Microbial Detection Array (LLMDA) and 16S rDNA sequencing. High growth outcomes were associated with several gut microbiome characteristics, such as increased bacterial diversity, increased Bacteroides pectinophilus, decreased Mycoplasmataceae species diversity, higher Firmicutes:Bacteroidetes ratios, increased relative abundance of the phylum Spirochaetes, reduced relative abundance of the family Lachnospiraceae, and increased Lachnospiraceae species C6A11 and P6B14. Overall, this study identifies gut microbiomes associated with improved outcomes in PRRS vaccinated pigs following a polymicrobial respiratory challenge and provides evidence towards the gut microbiome playing a role in PRRS vaccine efficacy.

Association of wild-type PRRSV detection patterns with mortality of MLV-vaccinated growing pig groups

PRRS is a viral disease characterized by increasing reproduction losses in breeding herds and worsening performance of growing pigs, which leads to a considerable economic impact.A better understanding of the differences in the infection patterns of the virus in growing pig batches would help to develop cost-effective surveillance methods and disease control and elimination programs. Therefore, field studies documenting growing pig productivity according to the patterns of wild-type-PRRSV (wt-PRRSV) detection in the field are needed. This study was conducted with the objectives to (1) characterize patterns of wt-PRRSV-1 and wt-PRRSV-2 RNA detection over time in modified-live virus (MLV)-vaccinated batches of growing pigs raised in pig-dense regions of the USA; (2) compare wean-to-finish mortality among batches of growing pigs characterized with the different patterns of wt-PRRSV-1 and wt-PRRSV-2 detection; and (3) compare wean-to-finish mortality among batches of growing pigs characterized with the different patterns of wt-PRRSV-2 detection and vaccinated with two different doses of PRRS MLV vaccine.Eighty-one batches of growing pigs were originated from PRRSV positive-stable and unstable sow farms and vaccinated with two different doses of PRRS MLV vaccine. All batches were monitored for wt-PRRSV by testing six oral fluids every three weeks from weaning to marketing. Diagnostics were conducted to detect wt-PRRSV-1 and wt-PRRSV-2 by RT-qPCR testing and ORF-5 region sequencing. K-means clustering analysis was applied to identify batches sharing similar patterns of wt-PRRSV detection over time. Regression analyses were used to compare mortality among batches of growing pigs characterized with different patterns of wt-PRRSV detection over time.Thirty-eight percent of batches were detected with wt-PRRSV-1 during the growth phase, with three different patterns of detection. Detection of wt-PRRSV-1 was not associated with significant increase on mortality. Ninety-one percent of batches were detected with wt-PRRSV-2 during the growth phase, with four different patterns of detection. Batches originated from PRRSV positive-unstable farms had highest mortality rate (p < 0.0001) and were characterized as Unstable wt-PRRSV-2 detection pattern. Batches characterized with Early wt-PRRSV-2 detection pattern had higher mortality than batches characterized by Mid, Late and No wt-PRRSV-2 detection during the growth phase (p < 0.0001). Batches with Mid wt-PRRSV-2 detection had higher mortality than batches characterized with Late wt-PRRSV-2 detection (p < 0.0124). Mortality rate of batches characterized with Unstable and Early wt-PRRSV-2 detection patterns was lower when pigs were vaccinated with two doses of PRRS MLV vaccine, compared to batches that received only one dose.Results presented in this study suggested that early wt-PRRSV exposure on pig populations was associated with higher wean-to-finish mortality. Additionally, results suggested that vaccination with two PRRS MLV doses was associated with lower mortality rate, when growing pig populations had early wt-PRRSV exposure.

Gilt Vaccination with a Mixed Administration of a PRRS MLV and a PPV1 Subunit Vaccine Protects against Heterologous PRRSV1 Infection and Prevents Detrimental Effects on Piglet Performance.

The efficacy of the combined administration of a porcine reproductive and respiratory syndrome (PRRS) modified live virus (MLV) vaccine and a porcine parvovirus 1 (PPV1) subunit vaccine in gilts was addressed in two experiments. Experiment A aimed to establish a 4-week onset of immunity (OOI). Gilts were randomly distributed in three treatment groups: non-vaccinated control animals (group 1), animals vaccinated with the combined vaccine (group 2), and a third group that consisted of animals vaccinated with the PRRS MLV vaccine alone (group 3). Four weeks after the first vaccination, gilts were challenged with a heterologous PRRS virus 1 (PRRSV1) and euthanized three weeks after. Besides this, experiment B pursued a 17-week duration of immunity (DOI). In this case, gilts were distributed in the same treatment groups, but for the third group, which consisted of non-vaccinated, non-challenged animals were used instead. For the DOI assessment, gilts were artificially inseminated 4 weeks after the first vaccination, challenged at day 90 of gestation, and followed up, together with their offspring, until day 20 post-farrowing. Serology and viremia post-challenge were determined in gilts from both experiments, while farrowing and piglet performance were only evaluated in experiment B. Overall, the combined vaccine helped to protect gilts from viremia post-challenge and, consequently, to prevent PRRS clinical symptoms and diminish the proportion of piglets infected congenitally or early in life. The combined vaccine also elicited a significant improvement in piglet survival rate and growth performance until weaning. The present results reveal efficacy and lack of interference of the mixed use of the tested vaccines against PRRSV1 infection, with at least 4-week OOI and 17-week DOI.

Assessment of immediate production impact following attenuated PRRS type 2 virus vaccination in swine breeding herds

BackgroundTo mitigate production impact of porcine reproductive and respiratory syndrome (PRRS) virus outbreaks, it has been common to preventively vaccinate swine breeding herds using PRRS modified live virus (MLV) vaccine. However, attenuated PRRS virus (PRRSv) may result negative impact on farm productivity. The objective of this study was to measure the immediate impact of PRRS type 2 MLV vaccine on breeding herd performance under field conditions. Eight PRRS-stable farms routinely mass vaccinating females with commercial PRRS MLV vaccines were enrolled on study. Vaccination dates were collected and weekly changes in abortions, neonatal losses, pre-weaning mortality, pigs weaned per sow, and wean-to-first-service interval were assessed for up to 6 weeks after each vaccination. A 6-week period prior to each vaccination was established as baseline. Statistical process control (SPC) analysis was conducted to detect significant productivity decreases after MLV interventions, on each farm, and a mixed regression model was used, at the aggregated data level, to assess the productivity change 6 weeks after PRRS MLV vaccinations, compared to baseline.ResultsOut of 65 herd-MLV vaccinations, SPC analysis detected increase on abortions 4 times (6.1%), on neonatal losses 7 times (10.7%), on pre-weaning mortality 2 times (3%), on wean-to-first-service interval 2 times (3%), and no change in total pigs weaned. On aggregated data analysis, there was no significant change in abortion rate, neonatal losses, number of pigs weaned per sow, and wean-to-first-service interval. However, there was an increase of 0.26% of pre-weaning mortality 2 weeks after vaccination compared to the baseline.ConclusionsUnder study conditions, individual PRRS-stable sow farms had experienced transient, and numerically small changes in productivity following PRRS type 2 MLV vaccination. There was a small increase of pre-weaning mortality 2 weeks after vaccination, but no evidence of significant production impact at aggregated data analysis for abortion rate, neonatal losses, pigs weaned per sow and wean-to-first-service interval.

Efficacy of Ingelvac PRRSandreg; Modified Live Virus Vaccine against Heterologous Porcine Reproductive and Respiratory Syndrome Virus Challenges

Porcine reproductive and respiratory syndrome (PRRS) is an infectious disease caused by the PRRS virus (PRRSV) and characterized by reproductive failure, respiratory disease and weight loss in swine. Two randomized, blinded vaccination-challenge studies evaluated the efficacy of Ingelvac PRRS® modified live virus (MLV) vaccine in protecting pigs from the virulent heterologous PRRSV isolates, restriction fragment length polymorphism (RFLP) 1-3-4 and 1-7-4. In separate challenge studies, pigs were vaccinated on Day 0 with Ingelvac PRRS MLV or placebo ‘challenge control’ and challenged on Day 28 with PRRSV 1-3-4 or 1-7-4. In the 1-3-4 challenge study, pigs vaccinated with Ingelvac PRRS MLV demonstrated significantly lower median viraemia (area-under-the-curve for Day 28–42 [AUC28–42]; P<0.0001) compared with unvaccinated controls. Vaccinated pigs also had significantly higher average daily weight gain (ADWG) than unvaccinated controls (P<0.0001). At Day 42, vaccinated pigs had significantly lower least square mean lung lesion scores than unvaccinated controls (P<0.001). Mortality was significantly higher with challenge control (61%) than with Ingelvac PRRS MLV (15%; P<0.01). In the 1-7-4 challenge study, significantly lower AUC28–42 viraemia levels were observed with Ingelvac PRRS MLV compared with challenge control (P=0.031). Median rectal temperatures were significantly lower with Ingelvac PRRS MLV than with challenge controls at Days 29 and 42 (P<0.01 for both). Pigs vaccinated with Ingelvac PRRS MLV had significantly higher ADWG during the challenge phase (P<0.05) and significantly lower least square mean lung lesion scores at Day 42 compared with unvaccinated controls (P<0.05). These data indicate that Ingelvac PRRS MLV provides heterologous protection against two relatively new and particularly virulent PRRSV field strains responsible for a growing number of infections in the US.

Effects of sample handling on the detection of porcine reproductive and respiratory syndrome virus in oral fluids by reverse-transcription real-time PCR.

We evaluated effects of handling procedures on detection of porcine reproductive and respiratory syndrome virus (PRRSV) in oral fluids (OFs) by reverse-transcription real-time PCR (RT-rtPCR). The experiments were conducted using a composite sample of PRRSV-positive OF collected from 5-wk-old pigs vaccinated 15 d earlier with a modified-live PRRSV vaccine. Five pre-extraction sample-handling steps and all combinations thereof were evaluated: 1) thaw temperature (4°C or 25°C); 2) sample diluent (1:1 dilution with nuclease-free water or guanidinium thiocyanate-phenol); 3a) sonication of the sample (yes or no); 3b) temperature (4°C or 25°C) at which step 3a was conducted; and 4) temperature at which the sample was maintained after step 3b and until RNA extraction was initiated (4°C or 25°C). All combinations of the 5 sample-handling steps (i.e., 32 unique treatments) were tested in a completely randomized factorial design with 4 replicates and 1 negative control for each treatment. The entire experiment was repeated on 5 separate days to produce a total of 800 PRRSV RT-rtPCR results. Binary (positive or negative) data were analyzed by logistic regression and results (Ct) were analyzed using a generalized linear model. Overall, 1 false-positive result was observed among 160 negative controls (99.4% specificity), and 85 false-negative results were observed among the 640 known-positive samples (86.7% sensitivity). The most significant factor affecting test outcome was thaw temperature (4°C or 25°C); samples thawed at 4°C had higher positivity rate (94% vs. 80%, p < 0.0001) and lower Ct (36.2 vs. 37.5, p < 0.0001).

Long duration of immunity against a type 1 heterologous PRRS virus challenge in pigs immunised with a novel PRRS MLV vaccine: a randomised controlled study

BackgroundPorcine reproductive and respiratory syndrome virus (PRRSV) is widespread in commercial pig farms worldwide, and has a significant cost to the swine industry. Herd owners need a vaccine that will confer long-lasting immunity to prevent PRRSV infection and transmission. The studies described here evaluated duration of immunity conferred by a European-derived PRRS (isolate 94,881) modified live virus (MLV) vaccine, Ingelvac PRRSFLEX® EU, at 20, 24, and 26 weeks post-vaccination. Primary endpoints were the assessment of gross and histological lung lesions and viral RNA load in lung tissue 10 days following heterologous PRRSV challenge. Secondary endpoints included clinical observations, average daily weight gain (ADWG) and viral RNA load in serum 10 days post-challenge. Three blinded, vaccination-challenge efficacy studies were performed using separate cohorts of pigs (n = 56 per study). Pigs received either Ingelvac PRRSFLEX® EU (Group 1) or placebo (Groups 2 and 3). Groups 1 and 2 were subsequently challenged with heterologous European PRRSV isolate 205,817 at 20, 24 or 26 weeks post-vaccination.ResultsMean gross lung lesion scores were significantly lower in Group 1 than in Group 2 at 24 and 26 weeks (p < 0.0001), but not at 20 weeks (p = 0.299). Significantly lower mean histological lung lesion scores were observed in Group 1 versus Group 2 at 20 (p = 0.0065), 24 (p < 0.0001) and 26 weeks (p < 0.0001). Mean viral RNA load in lung tissue was significantly lower in Group 1 than in Group 2 (p < 0.0001) at 20 (p < 0.0001), 24 (p < 0.0001) and 26 weeks (p < 0.0001). Cumulative viral RNA loads in serum during days 1–10 post-challenge were significantly lower in Group 1 than in Group 2 (p < 0.0001) in all studies. A significant increase in ADWG was observed in Group 1 compared with Group 2 at 20 weeks (p = 0.0027) and 24 weeks (p = 0.0004), but not at 26 weeks (p = 0.1041). There were no significant differences in clinical signs post-challenge in any study.ConclusionThese results suggest that Ingelvac PRRSFLEX® EU confers long-term immunity to European heterologous PRRSV, which is maintained up to 26 weeks after vaccination, corresponding to the expected lifespan of commercial pigs.

Efficacy evaluation of three modified-live virus vaccines against a strain of porcine reproductive and respiratory syndrome virus NADC30-like

Porcine reproductive and respiratory syndrome reproductive virus is a devastating pathogen causing tremendous economic losses to swine production worldwide. Emergence of novel and variant PRRSV strains always leads to variable protection efficacy of modified-live virus (MLV) vaccines. Prevalence of PRRSV NADC30-like recently emerging in China has brought about clinical outbreaks of the disease. In the present study, the pathogenicity of a NADC30-like strain CHsx1401 for piglets was analyzed, and the potential cross-protective efficacy of three MLV vaccines including two commercial MLV vaccines and an attenuated low pathogenic PRRSV against this virus was further evaluated in piglets. The NADC30-like CHsx1401 was shown to cause fever, respiratory clinical signs, and lung gross and microscopic lesions of the inoculated piglets, suggesting that this virus is moderate virulent for piglets. Vaccination of piglets with the MLV vaccines could not reduce the clinical signs and lung lesions, and was partially efficacious in the reduction of viral loads in sera upon NADC30-like CHsx1401 challenge, indicating that these three MLV vaccines provide extremely limited cross-protection efficacy against the NADC30-like virus infection. Additionally, Ingelvac PRRS MLV appeared to exert some beneficial efficiency in shortening the period of clinical fever and in improving the growth performance of the challenged pigs. Our findings give valuable guidance for the choice and use of PRRSV MLV vaccines to control NADC30-like virus infection in the field.

Efficacy of Fostera® PRRS modified live virus (MLV) vaccination strategy against a Thai highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) infection.

Recently, the Chinese highly pathogenic porcine reproductive and respiratory syndrome virus (PRRSV) (HP-PRRSV) belonging to lineage 8 causes severe symptom with high morbidity and high mortality rates to the Asian pig industry. A recent study showed that pigs immunized with Fostera® PRRS modified live virus (MLV) of lineage 8 could provide a degree of protection against a Vietnamese HP-PRRSV infection. It should be noted that PRRSV commonly found after weaning causes porcine respiratory disease complex (PRDC). Vaccination strategy should be evaluated in each farm scenario. Eighty-one PRRSV-free piglets obtained from a PRRS-free herd were divided into two experiments with the major difference of infection timing after vaccination, 42days in experiment 1 (n = 42) and 28days in experiment 2 (n = 39). Each experiment had similar protocol containing three groups including a negative control, unvaccinated challenged, and vaccinated challenged groups. Pigs in vaccination groups were immunized with Fostera® PRRS MLV vaccine at 3weeks of age. Then, unvaccinated challenged and vaccinated challenged groups were intranasally inoculated with a Thai HP-PRRSV (10PL01). Vaccinated challenged pigs showed significantly lower levels of mean rectal temperatures, clinical severity, lung lesion scores, and viral titers in serum and lung tissue compared to the unvaccinated challenged pigs (p < 0.05). Vaccinated challenged pigs had higher survival rate than those of unvaccinated challenged pigs in both experiments. It should be noted that pigs challenged 42days after vaccination showed a better performance than pigs challenged 28days after vaccination. In conclusion, Fostera® PRRS MLV vaccine was able to improve the survival rate against the Thai HP-PRRSV infection in both 42- and 28-day vaccination-to-infection protocols.

Modified-live PRRSV subtype 1 vaccine UNISTRAIN® PRRS provides a partial clinical and virological protection upon challenge with East European subtype 3 PRRSV strain Lena.

BackgroundWestern European porcine reproductive and respiratory syndrome virus (PRRSV) strains cause limited and mild clinical signs whereas more virulent strains are circulating in Eastern Europe. The emergence of such highly virulent strains in Western Europe might result in severe clinical problems and a financial disaster. In this context, the efficacy of the commercial modified-live PRRSV subtype 1 vaccine UNISTRAIN® PRRS was tested upon challenge with the East European subtype 3 PRRSV strain Lena.ResultsThe mean duration of fever was shortened and the number of fever days was significantly lower in vaccinated pigs than in control pigs. Moreover, a lower number of vaccinated animals showed fever, respiratory disorders and conjunctivitis. The mean virus titers in the nasal secretions post challenge (AUC) were significantly lower in the vaccinated group than in the control group. The duration of viremia was slightly shorter (not significantly different) in the vaccinated group as compared to the control group.ConclusionsVaccination of pigs with the modified-live vaccine UNISTRAIN® PRRS provides a partial clinical and virological protection against the PRRSV subtype 3 strain Lena.

The Attenuation Phenotype of a Ribavirin-Resistant Porcine Reproductive and Respiratory Syndrome Virus Is Maintained during Sequential Passages in Pigs.

In a previous study, ribavirin-resistant porcine reproductive and respiratory syndrome virus (PRRSV) mutants (RVRp13 and RVRp22) were selected, and their resistance against random mutation was shown in cultured cells. In the present study, these ribavirin-resistant mutants were evaluated in terms of their genetic and phenotypic stability during three pig-to-pig passages in comparison with modified live virus (MLV) (Ingelvac PRRS MLV). Pigs challenged with RVRp22 had significantly lower (P< 0.05) viral loads in sera and tissues than pigs challenged with MLV or RVRp13 at the first passage, and the attenuated replication of RVRp22 was maintained until the third passage. Viral loads in sera and tissues dramatically increased in pigs challenged with MLV or RVRp13 during the second passage. Consistently, all five sequences associated with the attenuation of virulent PRRSV in RVRp13 and MLV quickly reverted to wild-type sequences during the passages, but two attenuation sequences were maintained in RVRp22 even after the third passage. In addition, RVRp22 showed a significantly lower (P< 0.001) mutation frequency in nsp2, which is one of the most variable regions in the PRRSV genome, than MLV. Nine unique mutations were found in open reading frames (ORFs) 1a, 2, and 6 in the RVRp22 genome based on full-length sequence comparisons with RVRp13, VR2332 (the parental virus of RVRp13 and RVRp22), and MLV. Based on these results, it was concluded that RVRp22 showed attenuated replication in pigs; further, because of the high genetic stability of RVRp22, its attenuated phenotype was stable even after three sequential passages in pigs. PRRSV is a rapidly evolving RNA virus. MLV vaccines are widely used to control PRRS; however, there have been serious concerns regarding the use of MLV as a vaccine virus due to the rapid reversion to virulence during replication in pigs. As previously reported, ribavirin is an effective antiviral drug against many RNA viruses. Ribavirin-resistant mutants reemerged by escaping lethal mutagenesis when the treatment concentration was sublethal, and those mutants were genetically more stable than parental viruses. In a previous study, two ribavirin-resistant PRRSV mutants (RVRp13 and RVRp22) were selected, and their higher genetic stability was shown in vitro Consequently, in the present study, both of the ribavirin-resistant mutants were evaluated in terms of their genetic and phenotypic stability in vivo RVRp22 was found to exhibit higher genetic and phenotypic stability than MLV, and nine unique mutations were identified in the RVRp22 genome based on a full-length sequence comparison with the RVRp13, VR2332, and MLV genomes.