BackgroundPorcine reproductive and respiratory syndrome virus (PRRSV) causes devastating disease characterized by reproductive failure and respiratory problems in the swine industry. To understand the recent prevalence and genetic diversity of field PRRSVs in the Republic of Korea, open reading frames (ORFs) 5 and 7 of PRRSV field isolates from 631 PRRS-affected swine farms nationwide in 2013–2016 were analyzed along with 200 Korean field viruses isolated in 2003–2010, and 113 foreign field and vaccine strains.ResultsKorean swine farms were widely infected with PRRSVs of a single type (38.4 and 37.4% for Type 1 and Type 2 PRRSV, respectively) or both types (24.2%) with up to approximately 83% nucleotide sequence similarity to prototype PRRSVs (Lelystad or VR2332). Phylogenetic analysis based on the ORF5 nucleotide sequence revealed that Korean Type 1 field isolates were classified as subgroups A, B, and C under subtype 1, while Korean Type 2 field isolates were classified as lineages 1 and 5 as well as three Korean lineages (kor A, B, and C) with the highest infection prevalence in subgroup A (50.5%) and lineage 5 (15.3%) for Type 1 and Type 2 PRRSV, respectively, among ORF5-positive farms. In particular, the lineages kor B and C were identified as novel lineages in this study, and lineage kor B comprised only the field viruses isolated from Gyeongnam Province in 2014–2015, establishing regionally unique genetic characteristics. It has also recently been confirmed that commercialized vaccine-like viruses (subgroup C) of Type 1 PRRSV and NADC30-like viruses of Type 2 PRRSV (lineage 1) are spreading rapidly in Korean swine farms. The Korean field viruses were also expected to be antigenically variable as shown in the high diversity of neutralizing epitopes and N-glycosylation sites.ConclusionsThis up-to-date information regarding recent field PRRSVs should be taken into consideration when creating strategies for the application of PRRS control measures, including vaccination in the field.

Classical swine fever virus (CSFV) is highly contagious, and fatal to infected pigs. Vaccines against CSFV have been developed from attenuated or modified live viruses. These vaccines are effective for immunization of animals, but they are associated with problems such as the accidental spreading of viruses to animals in the field, and with barriers to trade following vaccination. Here, we report the generation of transgenic Nicotiana benthamiana plants for large-scale, cost-effective production of E2 fusion protein for use as a recombinant vaccine against CSFV in pigs. Transgenic N. benthamiana plants harboring an intergenic, single-copy insertion of a chimeric gene encoding E2 fusion protein had high levels of transgene expression. For large-scale production of E2 fusion protein from leaf tissues, we developed a protein-purification protocol consisting of cellulose-binding domain (CBD)–cellulose-based affinity purification and size-exclusion gel-filtration chromatography. E2 fusion proteins showed high immunogenicity in piglets and provided protection against CSFV challenge. The CBD in the E2 fusion protein was also highly immunogenic. These results suggest that plant-produced recombinant E2 fusion proteins can be developed into cost-effective vaccines against CSFV, with the CBD as a marker antigen to differentiate between vaccination and natural infection.

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In Jeju island of South Korea, a classical swine fever (CSF) non-vaccinated region, many pig farmers insisted on abortion and stillbirth in pregnant sows and high mortality of suckling/weaning piglets by circulating CSF virus from 2014 to 2018. We investigated whether CSF viruses isolated from pigs in Jeju Island (Jeju LOM) have recovered their pathogenicity by conducting experiments using pregnant sows and specific pathogen-free (SPF) pigs. The CSF modified live LOM vaccine (MLV-LOM) and Jeju LOM strains induced abortion and stillbirth in pregnant sows. Viral antigens were detected in the organs of fetuses and stillborn piglets in the absence of specific pathological lesions associated with the virulent CSF virus in both groups (MLV-LOM and Jeju LOM strain). However, antigen was detected in one newborn piglet from a sow inoculated with a Jeju LOM strain, suggesting that it may cause persistent infections in pigs. SPF pigs inoculated with the MLV-LOM or Jeju LOM strains were asymptomatic, but virus antigen was detected in several organ and blood samples. Virus shedding in both groups of animals was not detected in the feces or saliva until 21 days post inoculation. The serum concentration of the three major cytokines, IFN-α, TNF-α, and IL-10, known to be related to lymphocytopenia, were similar in both groups when the MLV-LOM or Jeju LOM strains were inoculated into SPF pigs. In conclusion, Jeju LOM strains exhibited most of the characteristics of the MLV-LOM in pigs and resulted in the same adverse effects as the MLV-LOM strain.

Atypical porcine pestivirus (APPV), currently classified as pestivirus K, causes congenital tremor (CT) type A-II in piglets. Eighteen APPV strains were identified from 2297 South Korean wild boars captured in 2019. Phylogenetic analysis of the structural protein E2 and nonstructural proteins NS3 and Npro classified the APPV viruses, including reference strains, into Clades I, II and III. Clade I was divided into four subclades; however, the strains belonging to the four subclades differed slightly, depending on the tree analysis, the NS3, E2, and Npro genes. The maximum-likelihood method was assigned to South Korean wild boar APPV strains to various subclades within the three trees: subclades I.1 and I.2 in the E2 tree, subclade I.1 in the Npro tree, and subclades I.1 and I.4 in the NS3 ML tree. In conclusion, APPV among South Korean wild boars belonging to Clade I may be circulating at a higher level than among the South Korean domestic pig populations.

Toxoplasma gondii and Neospora caninum are closely related protozoan parasites, they share many common hosts, and can cause neurological diseases in dogs. Dogs can have close contacts with humans and livestock and therefore they can act as reservoirs of these parasites. The aim of this study was to survey the seroprevalence of antibodies against T. gondii and N. caninum and their co-infection rate in dogs in Korea. In total, sera from 553 domestic dogs were collected from different breeds, sexes, and ages of dogs from nine provinces across the country of Korea during 2006 and 2007. The presence of antibodies against T. gondii and N. caninum was analyzed using the latex agglutination test (LAT) with a cut-off value of 1:32, and the indirect fluorescent antibody test (IFAT) using a serum titer of 1:100. In the total dog population, 71 (12.8%) dogs were positive for anti-T. gondii antibodies and only 20 (3.6%) were positive for anti-N. caninum antibodies. Relatively higher seropositive frequencies of antibodies against T. gondii (20.1%) and N. caninum (4.9%) were detected in the dog population from the Gyeonggi. A higher proportion of animals seropositive for anti-T. gondii antibodies was found in stray dog populations as compared to household dog populations: 18.5% (59/319) vs 5.1% (12/234), respectively. The Chi-square tests revealed significant differences in the seropositive frequencies of antibodies against T. gondii between stray and household dogs in the total population (p<0.0001), and in dogs from the Gyeonggi (p<0.01). No significant differences were observed for the presence of antibodies against T. gondii or N. caninum when compared across the sex or age (p>0.05). The first serological survey on antibodies against both T. gondii and N. caninum parasites across the entire country showed that co-infection was not common in these canine populations with a seropositive level of 0.72%. The significantly higher positive frequency of T. gondii antibodies in stray dogs in both, Gyeonggi and in the total dog populations suggests that further investigation on the seroprevalence of parasites should focus on stray dogs.

Gerbils (Meriones unguiculatus) were inoculated intraperitoneally (i.p.) with Neospora caninum tachyzoites to examine parasite distribution and histological lesions at different time points over a 9-day period of infection. Gerbils were sacrificed 12 h post-infection (PI), then daily intervals up to day 9 PI. The parasite was detected by PCR assay targeting the Nc5 sequence of N. caninum. The parasite was not found in any organs until day 5 PI, however, from day 8 PI onwards, they were detected in all the organs examined as demonstrated by PCR. The first target organs in acute N. caninum infection were liver, spleen, and kidney, but not the blood as was expected. Histologic lesions were detected in the liver and spleen only, no lesions were found in other organs examined until the end of the experiment. Notably, the focal miliary hepatitis was observed in the liver of infected gerbils just after 1 day post-inoculation, whereas splenic lesions were not found until day 5 PI. These results reinforce the applicability of gerbils as a suitable model of acute neosporosis and provide new insights into the response of gerbils to N. caninum intraperitoneal infection.