Porcine reproductive and respiratory syndrome (PRRS) is an endemic disease in the Republic of Korea. Surveillance of PRRS virus (PRRSV) types is critical to tailor control measures. This study collected 5062 serum and tissue samples between 2018 and 2022. Open reading frame 5 (ORF5) sequences suggest that subgroup A (42%) was predominant, followed by lineage 1 (21%), lineage 5 (14%), lineage Korea C (LKC) (9%), lineage Korea B (LKB) (6%), and subtype 1C (5%). Highly virulent lineages 1 (NADC30/34/MN184) and 8 were also detected. These viruses typically mutate or recombine with other viruses. ORF5 and non-structural protein 2 (NSP2) deletion patterns were less variable in the PRRSV-1. Several strains belonging to PRRSV-2 showed differences in NSP2 deletion and ORF5 sequences. Similar vaccine-like isolates to the PRRSV-1 subtype 1C and PRRSV-2 lineage 5 were also found. The virus is evolving independently in the field and has eluded vaccine protection. The current vaccine that is used in Korea offers only modest or limited heterologous protection. Ongoing surveillance to identify the current virus strain in circulation is necessary to design a vaccine. A systemic immunization program with region-specific vaccinations and stringent biosecurity measures is required to reduce PRRSV infections in the Republic of Korea.

As PCV2d infection has been continuously reported in swine farms in which pigs were vaccinated with PCV2a- or 2d-based vaccines, we attempted to develop a novel vaccine using a PCV2d-based capsid to enhance its protective efficacy. In this study, recombinant virus-like particles (VLPs) of rPCV2a, rPCV2b and rPCV2d were synthesized from the capsid proteins of PCV2a, PCV2b and PCV2d field isolates, respectively. A cross-neutralization assay between the VLPs induced antisera and the field isolates demonstrated the broad cross-neutralizing activities of the rPCV2d-induced antisera. Then, the protective efficacy of rPCV2d as a vaccine candidate was investigated in commercial pigs by rPCV2d vaccination and a single- or dual-challenge infection using a PCV2d strain and a type 1 PRRSV strain. High levels of anti-PCV2d IgG and neutralizing antibodies were induced 3 weeks after vaccination. After the challenge infection, the average ADWG values of the vaccinated group were higher than those of the unvaccinated group. None or a significantly low amount of (p < 0.05) reduced PCV2 genomic DNA was found in the blood, saliva and tissues of the vaccinated pigs, when compared to the unvaccinated group. Moreover, macroscopic and microscopic lesions in the tissues were significantly (p < 0.05) reduced in the vaccinated groups. This study therefore suggests that rPCV2d may be highly useful for the control of diverse field genotypes.

Porcine circovirus (PCV) 2d is a common genotype in South Korea, and the cross-protective ability of PCV2a-based vaccines has been reported recently. In this study, a PCV2d vaccine candidate was selected, and its protective efficacy against the PCV2d isolate was evaluated. From 2016 to 2020, 234 PCV2d isolates were phylogenetically analyzed using open reading frame 2 (ORF2) sequences and classified into four subgroups: PCV2d-1, PCV2d-2, PCV2d-3, and PCV2d-4. Except for PCV2d-4, which consisted of ungrouped isolates, the three subgroups showed distinct differences at amino acid positions 53 and 169 in the ORF2. The detection rates of PCV2d-1, PCV2d-2, and PCV2d-3 were 36.5, 37.4, and 3.7%, respectively, and representative isolates were selected from each subgroup (QIA244, QIA126, and QIA169, respectively). In the neutralization assay, QIA244 showed the lowest neutralization efficiency among the three PCV2a-based vaccines, whereas the virus-like particles of QIA244 (rQIA244) provided broader protection against the three genotypes than did those of QIA126 and rQIA169. To further evaluate rQIA244 in pigs, the experimental groups were divided into rQIA244-vaccine (2dVac), commercial PCV2a-vaccine (2aVac), and no-vaccination (noVac) groups. The 2dVac effectively reduced the copy number of PCV2d in blood and tissues, as well as in tissue lesions, compared to the effect of 2aVac. Collectively, 2dVac provided by QIA244 ORF2 successfully demonstrated protective efficacy against the currently prevalent PCV2d in vitro neutralization and in vivo assays.

Commercially used porcine respiratory and reproductive syndrome (PRRS) modified live virus (MLV) vaccines provide limited protection with heterologous viruses, can revert back to a virulent form and they tend to recombine with circulating wild-type strains. Codon pair deoptimization (CPD) is an advanced method to attenuate a virus that overcomes the disadvantages of MLV vaccines and is effective in various virus vaccine models. The CPD vaccine against PRRSV-2 was successfully tested in our previous study. The co-existence of PRRSV-1 and -2 in the same herd demands protective immunity against both viruses. In this study, live attenuated PRRSV-1 was constructed by recoding 22 base pairs in the ORF7 gene of the E38 strain. The efficacy and safety of the CPD live attenuated vaccine E38-ORF7 CPD to protect against virulent PRRSV-1 were evaluated. Viral load, and respiratory and lung lesion scores were significantly reduced in animals vaccinated with E38-ORF7 CPD. Vaccinated animals were seropositive by 14 days post-vaccination with an increased level of interferon-γ secreting cells. In conclusion, the codon-pair-deoptimized vaccine was easily attenuated and displayed protective immunity against virulent heterologous PRRSV-1.

Porcine reproductive and respiratory syndrome virus (PRRSV) is major economic problem given its effects on swine health and productivity. Therefore, we evaluated the genetic stability of a codon pair de-optimized (CPD) PRRSV, E38-ORF7 CPD, as well as the master seed passage threshold that elicited an effective immune response in pigs against heterologous virus challenge. The genetic stability and immune response of every 10th passage (out of 40) of E38-ORF7 CPD was analyzed through whole genome sequencing and inoculation in 3-week-old pigs. E38-ORF7 CPD passages were limited to 20 based on the full-length mutation analysis and animal test results. After 20 passages, the virus could not induce antibodies to provide effective immunity and mutations accumulated in the gene, which differed from the CPD gene, presenting a reason for low infectivity. Conclusively, the optimal passage number of E38-ORF7 CPD is 20. As a vaccine, this may help overcome the highly diverse PRRSV infection with substantially enhanced genetic stability.

This study aimed to evaluate the efficacy of a virus-like particle (VLP) vaccine containing the open reading frame 2 of porcine circovirus type 2d (PCV2d) in a farm environment where natural infections associated with porcine circovirus-associated disease are endemic. The vaccine trial was conducted on three farms (H, M, and Y) with a history of infections including porcine reproductive and respiratory syndrome virus (PRRSV), PCV, Mycoplasma, and E. coli. Farm H, as well as farms M and Y, experienced natural PCV2 infection between 4 and 8 weeks post-vaccination (wpv), and 8 and 12 wpv, respectively. Viremia levels of all farms were significantly (p < 0.05) lower in vaccinated piglets than the control group after natural infection. In all farms, serum immunoglobulin G levels peaked at 8 wpv in the vaccinated groups, surpassing those in the control groups. Furthermore, neutralizing antibody titers were significantly (p < 0.05) higher in the vaccinated groups than the control groups in farms H and Y (0–8 wpv). However, there were no significant differences between the vaccinated and control group in neutralizing antibody titers of farm M (0–20 wpv). In terms of body weight, vaccinated piglets from all three farms showed significantly increased average weights at 12 wpv compared to the control groups. In conclusion, our study revealed noteworthy differences in viremia and body weight gain between vaccinated and control animals on three farms. As a result, this field trial of PCV2d VLP vaccine was successful in protecting piglets from natural PCV2 infection.