PRRS Vaccine Research Articles

Protection efficacy and the safety of the synergy between modified Bazhen powder and PRRSV modified-live virus vaccine against HP-PRRSV in piglets.

The highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) poses a significant threat to the global swine industry. Vaccination is a preventive measure against viral infections. However, the use of vaccines in livestock healthcare programs faces the challenge of safety and delayed immune responses. Earlier studies have shown the potential of modified Bazhen powder as an immunomodulator with significant biological properties, but its effect on vaccines against HP-PRRSV is yet to be studied. This study elucidated how modified Bazhen powder could improve the safety and efficacy of the conventional PRRSV vaccine by evaluating T-cell responses, antibody levels, clinical symptoms, levels of viremia, organ health, and cytokine production. The results revealed that the oral application of modified Bazhen powder in combination with PRRS vaccination improved both cellular and humoral immunity, accelerated viremia clearance, improved lung injury scores, and reduced viral load in the tonsils. The modified Bazhen powder also effectively reduced inflammatory responses following a PRRSV challenge. These findings further highlight the pharmacological properties of modified Bazhen powder as a potential oral immunomodulatory agent that could enhance vaccine efficacy and ensure broad-spectrum protection against HP-PRRSV in pigs.

Risk factors for antimicrobial use in Dutch pig farms: A cross-sectional study

BackgroundAntimicrobial use (AMU) has decreased significantly in Dutch pig farms since 2009. However, this decrease has stagnated recently, with relatively high AMU levels persisting mainly among weaners. The aim of this study was to identify farm-level characteristics associated with: i) total AMU and ii) use of specific antimicrobial classes. MethodsIn 2020, cross-sectional data from 154 Dutch pig farms were collected, including information on AMU and farm characteristics. A mixed-effects conditional Random Forest analysis was applied to select the subset of features that was best associated with AMU. ResultsThe main risk factors for total AMU in weaners were vaccination for PRRS in sucklings, being a conventional farm (vs. not), high within-farm density, and early weaning. The main protective factors for total AMU in sows/sucklings were E. coli vaccination in sows and having boars for estrus detection from own production. Regarding antimicrobial class-specific outcomes, several risk factors overlapped for weaners and sows/sucklings, such as farmer's non-tertiary education, not having free-sow systems during lactation, and conventional farming. An additional risk factor for weaners was having fully slatted floors. For fatteners, the main risk factor for total AMU was PRRS vaccination in sucklings. ConclusionsSeveral factors found here to be associated with AMU. Some were known but others were novel, such as farmer's tertiary education, low pig aggression and free-sow systems which were all associated with lower AMU. These factors provide targets for developing tailor-made interventions, as well as an evidence-based selection of features for further causal assessment and mediation analysis.

Contrasting PRRSV temporal lineage patterns at the individual farm, production system, and regional levels in Ohio and neighboring states from 2017 to 2021

Porcine reproductive and respiratory virus (PRRSV), one of the most significant viruses in the swine industry, has been challenging to control due to its high mutation and recombination rates and complexity. This retrospective study aimed to describe and compare the distribution of PRRSV lineages obtained at the individual farm, production system, and regional levels. PRRSV-2 (type 2) sequences (n = 482) identified between 2017 – 2021 were provided by a regional state laboratory (Ohio Department of Agriculture, Animal Disease Diagnostic Center (ODA-ADDL)) collected from swine farms in Ohio and neighboring states, including Indiana, Michigan, Pennsylvania, and West Virginia. Additional sequences (n = 138) were provided by one collaborating swine production system. The MUSCLE algorithm on Geneious Prime® was used to align the ORF5 region of PRRSV-2 sequences along with PRRSV live attenuated vaccine strains (n = 6) and lineage anchors (n = 169). Sequenced PRRSV-2 were assigned to the most identical lineage anchors/vaccine strains. Among all sequences (n = 620), 29.8% (185/620) were ≥ 98.0% identity with the vaccine strains, where 93.5% (173/185) and 6.5% (12/185) were identical with the L5 Ingelvac PRRS® MLV and L8 Fostera® PRRS vaccine strains, respectively, and excluded from the analysis. At the regional level across five years, the top five most identified lineages included L1A, L5, L1H, L1C, and L8. Among non-vaccine sequences with production system known, L1A sequences were mostly identified (64.3% – 100.0%) in five systems, followed by L1H (0.0% - 28.6%), L1C (0.0% – 10.5%), L5 (0.0% – 14.4%), L8 (0.0% – 1.3%), and L1F (0.0% – 0.5%). Furthermore, among non-vaccine sequences with the premise identification available (n = 262), the majority of sequences from five individual farms were either classified into L1A or L5. L1A and L5 sequences coexisted in three farms, while samples submitted by one farm contained L1A, L1H, and L5 sequences. Additionally, the lineage classification results of non-vaccine sequences were associated with their restriction fragment length polymorphism (RFLP) patterns (Fisher’s exact test, p < 0.05). Overall, our results show that individual farm and production system-level PRRSV-2 lineage patterns do not necessarily correspond to regional-level patterns, highlighting the influence of individual farms and systems in shaping PRRSV occurrence within those levels, and highlighting the crucial goal of within-farm and system monitoring and early detection for accurate knowledge on PRRSV-2 lineage occurrence and emergence.

An Expeditious Neutralization Assay for Porcine Reproductive and Respiratory Syndrome Virus Based on a Recombinant Virus Expressing Green Fluorescent Protein.

Due to the extensive genetic and antigenic variation in Porcine Reproductive and Respiratory Syndrome Virus (PRRSV), as well as its rapid mutability and evolution, PRRS prevention and control can be challenging. An expeditious and sensitive neutralization assay for PRRSV is presented to monitor neutralizing antibodies (NAbs) in serum during vaccine research. Here, a PRRSV expressing eGFP was successfully rescued with reverse genetics based on the infectious clone HuN4-F112-eGFP which we constructed. The fluorescent protein expressions of the reporter viruses remained stable for at least five passages. Based on this reporter virus, the neutralization assay can be easily used to evaluate the level of NAbs by counting cells with green fluorescence. Compared with the classical CPE assay, the newly developed assay increases sensitivity by one- to four-fold at the early antibody response stage, thus saving 2 days of assay waiting time. By using this assay to unveil the dynamics of neutralizing antibodies against PRRSV, priming immunity through either a single virulent challenge or only vaccination could produce limited NAbs, but re-infection with PRRSV would induce a faster and stronger NAb response. Overall, the novel HuN4-F112-eGFP-based neutralization assay holds the potential to provide a highly efficient platform for evaluating the next generation of PRRS vaccines.

Genome stability assessment of PRRS vaccine strain with new ARTIC-style sequencing protocol.

A tiling amplicon sequencing protocol was developed to analyse the genome sequence stability of the modified live PRRSV vaccine strain, Porcilis MLV. The backbone of the ARTIC-style protocol was formed by 34 individual primer pairs, which were divided into two primer pools. Primer pairs were designed to amplify 532 to 588 bp fragments of the corresponding genomic region. The amplicons are suitable for sequencing on Illumina DNA sequencers with available 600-cycle sequencing kits. The concentration of primer pairs in the pools was optimized to obtain a balanced sequencing depth along the genome. Deep sequencing data of three vaccine batches were also analysed. All three vaccine batches were very similar to each other, although they also showed single nucleotide variations (SNVs) affecting less than 1 % of the genome. In the three vaccine strains, 113 to 122 SNV sites were identified; at these sites, the minority variants represented a frequency range of 1 to 48.7 percent. Additionally, the strains within the batches contained well-known length polymorphisms; the genomes of these minority deletion mutants were 135 to 222 bp shorter than the variant with the complete genome. Our results show the usefulness of ARTIC-style protocols in the evaluation of the genomic stability of PRRS MLV strains.

Evaluation of the efficacy of two postweaning colibacillosis vaccines in a field herd with PRRS circulation during postweaning stage

Postweaning diarrhea (PWD) and PRRS are two major concerns in swine production, which association has not been consistently explored. In the current scenario of restrictions in the use of antibiotics and ZnO, vaccination is more relevant to control PWD, but PRRS virus circulation may compromise the immune protection conferred by postweaning colibacillosis vaccines. We evaluated the efficacy of two postweaning colibacillosis vaccines (parenteral and oral) in a commercial herd affected by an outbreak of PWD and with PRRS circulation in postweaning. Five groups were studied during the postweaning period: one control (Group 1) and four vaccinated: two with each postweaning colibacillosis vaccine administered alone (Groups 2 and 3) or with sow vaccination against PRRS (Groups 4 and 5). We evaluated the effects on piglet weight, average daily weight gain and in the percentage of piglets with diarrhea, its duration, lethality and mortality. PRRS viremia and anti-PRRS antibodies were evaluated by qPCR and ELISA. Regarding control group, colibacillosis vaccination generally improved most of the measured parameters; but significant improvements were only observed in Groups 4 and 5 (p < 0.05). Moreover, cases of diarrhea occurred at different ages: in Groups 2 and 3 the peak of cases occurred just after ZnO was removed from the feed compared to Group 1, while in Groups 4 and 5 no peak was observed. This suggests that postweaning colibacillosis vaccination may be compromised by the PRRS circulation. In PRRS endemic herds an effective protection against PWD through vaccination may require PRRS vaccination to obtain a better performance.

Assessment of the genetic changes of the attenuated Hanvet1.vn strain compared with original virulent 02HY strain of the porcine reproductive and respiratory syndrome virus

The attenuated porcine reproductive and respiratory syndrome virus (PRRSV) strain Hanvet1.vn was developed by Hanvet Pharmaceutical Co., Ltd. by inoculating the virulent strain 02HYon Marc-145 cells for 80 generations and used to produce PRRS vaccine. In this study, we published the results of sequencing, analyzing and comparing the genome of the attenuated PRRSV strain Hanvet1.vn compared with the original pathogenic strain 02HY. The genomes of strains Hanvet1.vn and 02HY have 8 reading frames, coding for 8 non-structural and structural proteins: NSP1a, NSP1b, GP2, GP3, GP4, GP5, MP, NP. After sequencing and translating into proteins, the gene sequence of each open reading frame (ORF) of strain Hanvet1.vn was compared with the sequence of pathogenic strain 02HY to find nucleotide and amino acid changes. The results showed that the Hanvet1.vn pathogenic strain genome (Genbank Accession KU842720) when compared with the pathogenic strain 02HY genome (Submission2490633) had89 nucleotide mutations that changed 51 amino acids in 7 ORFs and 7 proteins, respectively. Particularly, ORF6 encoding for the M protein is completely unchanged. The size of each reading frame is also exactly the same. It showed that there were no insertion and deletion (Indel) mutations in the ORFs of the attenuated strain after 80 generations of inoculation. There was a change in the genome that made the strain Hanvet1.vn become attenuated, but the gene encoding for the GP5 protein that induces the production of neutralizing antibodies only changed two nucleotides at position 471 (A-&gt;G), causing the TCA codon to turn into a TCG codon. This is a silent mutation and both codons code for the amino acid Serine (S). The second mutation at position 587 (A-&gt;T) causes Glutamine (Q) to transform into Leucine (L). However, this modification does not belong to the GP5 antigenic epitopes. In clonclusion, after 80 passages, despite changes occurred in genes of Hanvet1.vn strain for becoming an attenuated strain, the GP5 protein of the attenuated strain did not change its antigenic amino acids.

Phylogenetic analysis of porcine reproductive and respiratory syndrome virus in Vietnam, 2021.

The porcine reproductive and respiratory syndrome virus (PRRSV) causes more economic losses in the swine industry than any other virus. This study aimed to investigate the genetic diversity of PRRSV to assist in evaluating the effectiveness of PRRS vaccines. Twenty-eight samples from clinical cases were collected from 19 farms in seven provinces of Vietnam in 2021. Full-length PRRSV ORF5 genes from the 19 samples were amplified, sequenced, and compared to the corresponding sequences of referenced PRRSV strains from Genbank. The genetic analysis showed that 12 isolates were the highly pathogenic PRRSV subtype (HP—PRRSV) lineage 8, sublineage 8.7; six isolates were the classical North American PRRSV subtype (US-PRRSV), NADC-like group, lineage 1, sublineage 1.4, which were reported in Vietnam for the first time; and the final isolate was a vaccine-like strain. The field isolates of HP-PRRSV had relatively higher genetic diversity with US-PRRSV vaccine strains (84.0–94.5%) than HP-PRRSV vaccine strains (95.3–98.6%). Meanwhile, the six NADC-like isolates had low nucleotide similarity with US-PRRSV and HP-PRRSV vaccine strains (83.4–85.4% and 83.2–84.0%, respectively). Many amino acid substitutions were found in antigenic regions of GP5 involved in response to early antibody production, neutralizing antibodies, and viral immune evasion between these field strains and PRRSV vaccine strains. These findings provide insights into the molecular characteristics, genetic diversity, antigenicity, and evolution of PRRSV strains in Vietnam and postulate a compelling explanation for the limitations of current vaccination efforts.Supplementary InformationThe online version contains supplementary material available at 10.1007/s11262-022-01912-w.

Minor and major envelope proteins of PRRSV play synergistic roles in inducing heterologous neutralizing antibodies and conferring cross protection

High genetic diversity of porcine reproductive and respiratory syndrome virus (PRRSV) isolates is a major obstacle for the development of effective PRRS vaccines. A chimeric highly pathogenic PRRSV2 (HP-PRRSV2) strain containing the consensus sequence of ORF2-6 genes was constructed in our previous study, which could induce broadly neutralizing antibodies (bnAbs) and confer satisfied cross protection against virulent NADC30-like isolate. To further elucidate the roles of minor and major envelope proteins encoded by ORF2-4 and ORF5-6 genes in conferring cross protection, two chimeric HP-PRRSV2 strains (rJS-ORF2-4-CON and rJS-ORF5-6-CON) containing consensus sequences of ORF2-4 or ORF5-6 were constructed and rescued in this study. The rJS-ORF5-6-CON strain has similar replication efficiency as the backbone HP-PRRSV2 rJSTZ1712-12 virus, while rJS-ORF2-4-CON has significantly lower in vitro and in vivo replication efficiency comparing to rJS-ORF5-6-CON. Animal inoculation indicated that both rJS-ORF2-4-CON and rJS-ORF5-6-CON did not cause obvious clinical signs in piglets and could induce heterologous nAbs after immunization. Challenge with a virulent heterologous NADC30-like SD17-38 isolate showed that even though both immunized groups presented lower viremia, faster virus elimination, less fever and alleviated lung gross lesions when compared with the only challenged pigs, rJS-ORF2-4-CON and rJS-ORF5-6-CON could not confer enough cross protection. Considering the bnAbs and satisfied cross protection induced by the chimeric virus containing ORF2-6 consensus sequence, our results support that minor and major envelope proteins play synergistic roles in inducing broader nAbs and conferring better cross protection.

PSVI-14 Impact of Bacillus Subtilis and Bacillus Licheniformis on Weaning Weight and Subsequent Performance, Health and Immune Parameters When Challenged with PRRSV with and Without PRRS Vaccination

Abstract A study was conducted examining the impact of feeding BioPlus 2B to PRRSV naïve sows during gestation and lactation on litter performance and on post-weaning performance when challenged with PRRSV in vaccinated and unvaccinated pigs. A total of 108 sows were allotted by parity and received a control diet (C) or BioPlus 2B (B2B) supplemented diet beginning a minimum of 31 d prior to farrowing through weaning. Pig weight was measured at birth and 17 d of age. At weaning, a subset of 200 pigs from each treatment were transported to a nursery and randomly allotted within dietary treatment on day 1 to a control (-V) or Ingelvac PRRS MLV vaccine (+V), establishing treatments of C+V, C-V, B2B+V, and B2B-V. On d 21, all pigs were intramuscularly challenged with a 1-18-2 PRRS virus. Growth rate and feed intake were monitored, serological and immune measures were recorded on d 21, 28, 35 and 42, post-weaning and on d 42 gross lung lesions were recorded. Pigs from sows receiving B2B tended (P &amp;lt; 0.10) to weigh more at 17 d of age and 21 d post-weaning (Table 1). Prior to challenge, there were no performance differences among treatments. Post challenge, there was a V x B2B interaction with C+V, B2B+V and B2B-V pigs having higher ADG and FG (P &amp;lt; .05) compared with C-V. On day 42, pigs fed B2B displayed improved lung lesion scores (P &amp;lt; 0.05) regardless of vaccine status. PRRSV viremia, measured by PCR and qPCR decreased more quickly when pigs consumed B2B (P &amp;lt; 0.05). Additionally, B2B modulated PRRSV specific T-cell interferon gamma production (P &amp;lt; 0.05). In conclusion, pigs consuming B2B from sows fedB2B were heavier at weaning and when subjected to a PRRSV challenge post-weaning, grew faster, were more efficient, and had reduced gross lung lesions, likely due to changes in immune function.

Systemic Homologous Neutralizing Antibodies Are Inadequate for the Evaluation of Vaccine Protective Efficacy against Coinfection by High Virulent PEDV and PRRSV.

ABSTRACTG2 porcine epidemic diarrhea virus (G2 PEDV) and highly pathogenic porcine reproductive and respiratory syndrome virus 2 (HP-PRRSV2) are two of the most prevalent swine pathogens in China’s swine herds, and their coinfection occurs commonly. Several PED and PRRS vaccines have been utilized in China for decades, and systemic homologous neutralizing antibodies (shnAbs) in serum are frequently used to evaluate the protective efficacy of PED and PRRS vaccines. To develop a vaccine candidate against G2 PEDV and HP-PRRSV2 coinfection, in this study, we generated a chimeric virus (rJSTZ1712-12-S) expressing S protein of G2 PEDV using an avirulent HP-PRRSV2 rJSTZ1712-12 infectious clone as the viral vector. The rJSTZ1712-12-S strain has similar replication efficacies as the parental rJSTZ1712-12 virus. In addition, animal inoculation indicated that rJSTZ1712-12-S is not pathogenic to piglets and can induce shnAbs against both G2 PEDV and HP-PRRSV2 isolates after prime-boost immunization. However, passive transfer study in neonatal piglets deprived of sow colostrum showed that rJSTZ1712-12-S-induced shnAbs may only decrease PEDV and PRRSV viremia but cannot confer sufficient protection against dual challenge of high virulent G2 PEDV XJ1904-34 strain and HP-PRRSV2 XJ17-5 isolate. Overall, this study provides the first evidence that shnAbs confer insufficient protection against PEDV and PRRSV coinfection and are inadequate for the evaluation of protective efficacy of PED and PRRS bivalent vaccine (especially for the PED vaccine).IMPORTANCE Porcine epidemic diarrhea virus (PEDV) and porcine reproductive and respiratory syndrome virus (PRRSV) coinfection occurs commonly and can synergistically reduce feed intake and pig growth. Vaccination is an effective strategy utilized for PED and PRRS control, and systemic homologous neutralizing antibodies (shnAbs) in serum are commonly used for protective efficacy evaluation of PED and PRRS vaccines. Currently, no commercial vaccine is available against PEDV and PRRSV coinfection. This study generated a chimeric vaccine candidate against the coinfection of prevalent PEDV and PRRSV in China. The chimeric strain can induce satisfied shnAbs against both PEDV and PRRSV after prime-boost inoculation in pigs. But the shnAbs cannot confer sufficient protection against PEDV and PRRSV coinfection in neonatal piglets. To the best of our knowledge, these findings provide the first evidence that shnAbs confer insufficient protection against PEDV and PRRSV coinfection and are inadequate for evaluating PED and PRRS bivalent vaccine protective efficacy.

The Safety and Efficacy Evaluation of PRRS Attenuated Vaccine BG895 Strain in Experimental Pigs

From the PRRSV virulent strain BG8 isolated from a PRRSV-infected pig, using serial passage method in MARC-145 cell line, we have successfully obtained an attenuated strain in 95th passage, named as BG895, with high potential to be a vaccine candidate. In this study, we present the results of the safety and efficacy evaluation of BG895 against PRRSV in experimental pigs. Trial results of vaccine formula using strain BG895 have very high safety when inoculating 5 doses/animal and 10 doses/animal. Evaluation of immune response by ELISA method showed that, from 14 days post inoculation, anti-PRRSV antibodies were detected in the serum of all inoculated pigs in vaccine batches with the lowest S/P index of 1.50 ± 0.4 and the highest S/P was 2.36 ± 0.1 from 28 days post inoculation. The IPMA method showed that the antibody titer of the vaccine reached ≥ 1/160 in 100% of pigs from 21 days post inoculation andreached ≥ 1/640 in 100% of pigs from 28 days post inoculation, indicating that the vaccine was effective at protecting 100% of pigs from 28 days post inoculation. The protective effect of the vaccine was evaluated by the virulent challenge from 28 days post inoculation with 1 dose/animal compared with the control group. The results showed that compared with all pigs in the control group with typical clinical manifestations of Blue-ear disease, all inoculated pigs had normal body temperature and weight gain, besides, the S/P index increased from 1.65 ± 0.1 to the highest 2.99 ± 0.2; the average antibody titer was &gt;1/2560, and virus wasnot detected in nasal fluid by real-time RT-PCR from 7 days post challenge. These experimental results confirmed the safety and efficacy of the attenuated PRRS vaccine based on BG895strain.

ОСОБЕННОСТИ ДИАГНОСТИКИ И ТЕРАПИИ СТРЕПТОКОККОЗА СВИНЕЙ, ОСЛОЖНЕННОГО РРСС НА ПЛЕМЕННОЙ ФЕРМЕ

Pathological phenomena began to appear in April in piglets after weaning from sows and in piglets weighing up to 30 kg. Clinical signs: cough, diarrhea, vomiting, tremor, lameness and extensive eczema-like skin lesions (spots of hyperemia on the skin of the breast and abdominal part of the body, redness of the eyelids), palpation - increased tactile sensitivity. The disease often proceeded in a septic form and was accompanied by fever. A significant part of the diseased pigs died. Mortality among piglets with streptococcosis varied from 66.6 to 100%. Bacteriological and molecular genetic (PCR) diagnostics of streptococcosis and reproductive and respiratory syndrome were carried out. As a result of PCR with porcine brain samples, Streptococcus suis DNA was isolated in 100% of cases. In the study of PCR swabs from the vagina of sows, Streptococcus suis DNA was isolated in 79% of cases. The use of bacteriological and molecular genetic (PCR) diagnostics of streptococcosis and PRRS virus made it possible to establish an etiological diagnosis - streptococcosis in association with PRRS. For treatment, an antibacterial drug of the third generation cephalosporin group Naxcel (Naxcel) was used, containing ceftiofur, Enroxil 10% injection solution, containing enrofloxacin as an active substance, as well as oral veterinary drugs Trisulfon and Pulmokit. Pregnant sows and piglets after weaning, without signs of infection with the reproductive and respiratory syndrome virus, were treated with antibacterial drugs (Naxel, Enroxil, Trisulfon and Pulmokit), containing ceftiofur and enrofloxacin, which allowed to inactivate Streptococcus suis in the body of piglets. The use of specific prophylaxis of the reproductive and respiratory syndrome on pregnant sows and piglets after weaning, without signs of disease, using the Unistrain PRRS vaccine against PRRS, contributed to the improvement of the epizootic situation for bronchopulmonary diseases on the breeding farm. Comprehensive recreational activities made it possible to sanitize the livestock from bacterial infections, including streptococcosis.

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.

Polyplex nanomicelle delivery of self-amplifying RNA vaccine

Self-amplifying RNA (SaRNA) is a burgeoning platform that exploits the replication machinery of alphaviruses such as Venezuelan equine encephalitis (VEE) virus or Sindbis virus (SIN). SaRNA has been used for development of human vaccines, but has not been evaluated for porcine vaccine development. Porcine reproductive and respiratory syndrome virus (PRRSV) causes tremendous economic losses to the worldwide pork industry, but current vaccines trigger delayed neutralizing antibody response and confer only partial protection. Here we first compared two SaRNA systems based on VEE and SIN, and demonstrated that in vitro transcribed VEE-based SaRNA conferred prolonged reporter gene expression and RNA amplification in pig cells with low cytotoxicity, but SIN-based SaRNA imparted evident cytotoxicity and limited gene expression in pig cells. Transfection of VEE-based SaRNA that encodes the major PRRSV antigen dNGP5 (SaRNA-dNGP5) conferred persistent expression for at least 28 days in pig cells. We next complexed SaRNA-dNGP5 with the polyaspartamide block copolymer PEG-PAsp(TEP) to form polyplex nanomicelle with high packaging efficiency and narrow size distribution. The polyplex nanomicelle enabled sustained dNGP5 expression and secretion in vitro. Compared with the commercial PRRS vaccine, nanomicelle delivery of SaRNA-dNGP5 into animal models accelerated the induction of potent neutralizing antibodies with minimal side effects, and elicited stronger IL-4 and IFN-γ responses against homologous and heterologous PRRSV. These properties tackle the problems of current vaccines and implicate the potential of SaRNA-dNGP5 nanomicelle as an effective PRRS vaccine.

Chimeric HP-PRRSV2 containing an ORF2-6 consensus sequence induces antibodies with broadly neutralizing activity and confers cross protection against virulent NADC30-like isolate

Due to the substantial genetic diversity of porcine reproductive and respiratory syndrome virus (PRRSV), commercial PRRS vaccines fail to provide sufficient cross protection. Previous studies have confirmed the existence of PRRSV broadly neutralizing antibodies (bnAbs). However, bnAbs are rarely induced by either natural infection or vaccination. In this study, we designed and synthesized a consensus sequence of PRRSV2 ORF2-6 genes (ORF2-6-CON) encoding all envelope proteins based on 30 representative Chinese PRRSV isolates. The ORF2-6-CON sequence shared > 90% nucleotide identities to all four lineages of PRRSV2 isolates in China. A chimeric virus (rJS-ORF2-6-CON) containing the ORF2-6-CON was generated using the avirulent HP-PRRSV2 JSTZ1712-12 infectious clone as a backbone. The rJS-ORF2-6-CON has similar replication efficiency as the backbone virus in vitro. Furthermore, pig inoculation and challenge studies showed that rJS-ORF2-6-CON is not pathogenic to piglets and confers better cross protection against the virulent NADC30-like isolate than a commercial HP-PRRS modified live virus (MLV) vaccine. Noticeably, the rJS-ORF2-6-CON strain could induce bnAbs while the MLV strain only induced homologous nAbs. In addition, the lineages of VDJ repertoires potentially associated with distinct nAbs were also characterized. Overall, our results demonstrate that rJS-ORF2-6-CON is a promising candidate for the development of a PRRS genetic engineered vaccine conferring cross protection.

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.

A recombination between two Type 1 Porcine Reproductive and Respiratory Syndrome Virus (PRRSV-1) vaccine strains has caused severe outbreaks in Danish pigs.

Porcine reproductive and respiratory syndrome virus (PRRSV) is prevalent in Danish swine herds. In July 2019, PRRSV‐1 was detected in a PRRSV‐negative boar station and subsequently spread to more than 38 herds that had received semen from the boar station. Full genome sequencing revealed a sequence of 15.098 nucleotides. Phylogenetic analyses showed that the strain was a recombination between the Amervac strain (Unistrain PRRS vaccine; Hipra) and the 96V198 strain (Suvaxyn PRRS; Zoetis AH). The major parent was the 96V198 strain that spanned ORFs 1–2 and part of ORF 3 and the minor parent was the Amervac strain, which constituted the remaining part of the genome. The virus seems to be highly transmissible and has caused severe disease in infected herds despite a high level of genetic identity to the attenuated parent strains. The source of infection was presumable a neighbouring farm situated 5.8 km from the boar station.

The efficacy and performance impact of Fostera PRRS in a Vietnamese commercial pig farm naturally challenged by a highly pathogenic PRRS virus

Highly pathogenic porcine reproductive and respiratory syndrome virus (HP-PRRSV) is characterized by high fever, respiratory distress, and high mortality in pigs of all ages and has severely affected the Vietnam pork industry in recent years. The study was conducted to compare the efficacy, safety, and overall performance of a modified live PRRSV-2 vaccine (Fostera PRRS) to an existing PRRSV modified live vaccine on a farm with a recent history of HP-PRRSV-associated respiratory diseases. A total of 351 pigs were randomly allocated to three treatment groups: (i) vaccinated with Fostera PRRS at 1day of age (n = 118), (ii) vaccinated with Fostera PRRS (n = 118) at 21days of age, and (iii) vaccinated with Amervac PRRS (n = 115) at 21days of age. The Fostera PRRS vaccinated pigs had milder clinical symptoms, lower levels of HP-PRRSV viremia, fewer pathological changes in the lung, and higher body weight gain at the end of the study compared with the Amervac PRRS group. Vaccination of pigs with Fostera PRRS at 1day of age also significantly reduced viral loads in their blood (P < 0.05) and induced higher anti-PRRSV antibody titers (P < 0.01) compared with pigs vaccinated with Amervac PRRS at 21days of age. Fostera PRRS vaccination at 1day of age can be useful in protecting young piglets from early HP-PRRSV infection because the immunized pigs were marketed 20days earlier than their peers immunized at 21-day old as they reached the target market weight earlier in this study.

Use of processing fluid samples for longitudinal monitoring of PRRS virus in herds undergoing virus elimination

This was an observational study that prospectively followed 29 breeding herds for 65 weeks in the U.S.A. that became infected with porcine reproductive and respiratory syndrome virus (PRRSv). The herds operated in a four-week batch farrowing system and adopted a load-close-expose strategy using a modified-live virus vaccine to achieve PRRSv stability. The purpose of this study was to describe time to stability (TTS) based on RT-qPCR testing for PRRSv RNA on processing fluid samples in herds undergoing PRRSv elimination, after implementing herd closure and mass exposure to a PRRS modified-live virus (MLV) vaccine. For the purpose of this study, stability was defined as consistently producing PRRSv-negative pigs. Study herds were monitored until two consecutive piglet batches tested PRRSv RT-qPCR negative, then 30 due-to-wean piglet sera from the second batch were tested for PRRSv RNA by RT-qPCR. Once the farm re-opened, sera from incoming naïve gilts were tested for anti-PRRSv antibodies by ELISA at 30- and 60-days post-entry to confirm negative status to PRRSv. Day zero was the day of whole-herd exposure to a commercial PRRS vaccine virus. Twenty-eight of 29 herds (96.55%) achieved TTS within the study period. TTS ranged from 18 to 55 weeks with a median of 27 weeks. Serum from due-to-wean piglets was collected on 28 farms, of which 26 (92.85%) obtained PRRSv RT-qPCR-negative results on the first collection. At the end of the observational period, 16 sow farms successfully re-introduced PRRSv-naïve gilts with no detected serologic response. In conclusion, the median time to achieve TTS in breeding herds being operated in a four-week batch farrowing system undergoing PRRSv elimination using load-close-expose with attenuated virus vaccine was 27 weeks. Also, processing fluid-based monitoring of breeding herds under PRRS elimination was practical and reliable to assess PRRSv stability.