Canine parvovirus-2 (CPV-2) has been reported worldwide as a major pathogen associated with acute hemorrhagic enteritis. The disease is a major infectious cause of death, particularly in young dogs. The earliest type of CPV-2 was replaced with three main subspecies, CPV-2a, CPV-2b, and CPV-2c, within a few years. Vaccination is carried out regularly, but the emergence of antigenic variants and the influence of maternal antibodies have limited the efficacy of commercial vaccines. New vaccines, such as the subunit vaccine, have been developed for alternative, safe, and effective vaccination. The baculovirus expression vector system (BEVS) is an excellent eukaryotic expression system with a high-level expression of foreign proteins and the ability of post-translational modification. Therefore, it is used widely to produce recombinant protein and subunit vaccines. In this study, the VP2 protein of CPV-2b cloned in the gateway vector system was generated using a baculovirus expression system in Spodoptera frugiperda (SF9) insect cells. Hemagglutination assay (HA) titers (24) were obtained, and the expression was detected in 6-His tagged VP2 and monoclonal antibody (mAb) against CPV-2 by western blotting. The VP2 protein of CPV-2b expressed in this study may provide a basis for a clinical diagnosis and vaccination applications for CPV-2.

Influenza A viruses (IAVs) are members of the family Orthomyxoviridae and genus Orthomyxovirus. Avian and mammalian species are the host of IAVs, which includes humans and dogs. Canine influenza virus (CIV) is an emerging pathogen that causes severe and acute respiratory diseases in dogs. This study monitored the antigen and antibody against CIV in dogs in the Republic of Korea (ROK) from 2016 to 2021. One thousand and seventy-two nasal swabs and 1,545 blood samples were collected from animal hospitals and animal shelters. Five nasal swabs in 2017 and seven in 2018 from stray dogs were positive for CIV according to RT-PCR. The prevalence of H3N2 CIV ranged from 9.5% to 24.8%, according to the hemagglutination inhibition (HI) assay. On the other hand, none of the serum samples from 2018 to 2021 showed seropositivity against the avian H5, H7, and H9 viruses. The HI titers for H3N2 ranged from 16 to 512. The distribution of HI titer 16–32 was 57.6% in seropositive samples. The pet dogs were vaccinated against CIV, but the stray and military dogs were unvaccinated. In 2017 and 2018, the seroprevalence of CIV in stray dogs was higher than in the other years, and viral RNA was detected in nasal swabs. It may mean previous exposure of stray dogs to CIV. With the increasing number of pet dogs and the close contact between humans and dogs, canines could serve as an intermediate host for transmitting IAVs to humans. Therefore, continuous surveillance of CIV is needed for public health and the potential emergence of novel zoonotic viruses.

The canine parvovirus (CPV) causes clinical signs, such as severe enteritis, dehydration, diarrhea, vomiting, leukopenia, and hair loss, which may lead to death. Vaccination is still the most important approach, as no specific treatment exists to prevent CPV. Monoclonal antibodies are valuable tools to study the pathogenic mechanisms of CPV and develop effective diagnostic reagents and pharmaceuticals. In this study, two monoclonal antibodies (MAbs) against CPV-2a were obtained through hybridoma technology by fusing myeloma cells and B cells from the spleens of mice immunized with CPV type 2a (CPV-2a). Two MAbs (CPV-330 and CPV-620) were studied on the reactivity of vaccine (CPV-2a) and field strains (CPV-new 2a, -2b, and -2c) by indirect immunofluorescence (IFA), hemagglutination inhibition test (HI), virus neutralization test (VN), and inhibition of virus growth test. Two MAbs showed similar antibody titers for HI and VN. On the other hand, CPV-330 inhibited the viral replication in Crandell-Rees Feline Kidney (CRFK) cells better than CPV-620. These CPV MAbs may provide valuable biological reagents to study the CPV pathogenic mechanisms and work as therapeutic antibodies.

Natural killer (NK) cells have cytotoxic effects on tumor cells and viral pathogens. NK cell-derived exosomes (NK-exosomes) also express typical NK cell markers and cytotoxic molecules, therefore, exert anti-tumor and immune homeostatic activities. In this study, canine NK-exosomes separated from cytotoxic NK cell supernatant carried specific markers such as CD81, Alix, and Perforin 1. We examined the anti-tumor effects of NK-exosomes in an experimental murine model using the canine mammary carcinoma cells, REM134. REM134 cells were xenografted of mammary fat pad of mice. CD133, Bmi-1, MMP-3, IL-6, TNF-α, and PCNA are useful as a molecular marker for tumorigenesis and metastasis. The treatment of canine NK-exosomes inhibited tumor growth and significantly (p<0.01) downregulated the expression of Bmi-1, MMP-3, IL-6, TNF-α, and PCNA in REM134-treated mice. Also, the expression of CD133, potent cancer stem cell marker, was significantly downregulated in the canine NK-exosomes-treated mice compared with that of the tumor group. Collectively, these results suggested that canine NK-exosomes has a potential capacity for regulation of cancer progression and metastasis against canine mammary carcinoma.

Mesenchymal stem cells (MSCs) are multipotent cells capable of replicating as undifferentiated cells, and thus hold therapeutic implications in field of regenerative medicine and reproductive biotechnology. In the present study, we compared the stem cell properties of bovine ear skin tissue (ESK)- and nasal mucosa (NM)-derived MSCs. Bovine ESK-MSCs and NM-MSCs were successfully isolated by collagenase digestion and maintained proliferative capacity during the 20 consecutive passages. Both ESK-MSCs and NM-MSCs showed similar morphology and expressed common cell surface markers (CD29, CD44, CD90, and CD105). Also, we compared differentiation potentials of bovine ESK-MSCs and NM-MSCs into osteogenic, adipogenic, and chondrogenic lineages through specific staining and quantitative real-time RT-PCR. As results, bovine ESK-MSCs and NM-MSCs could differentiate into mesodermal cell lineages. However, bovine ESK-MSCs and NM-MSCs exhibited difference in expression of differentiation-related specific markers. Specifically, NM-MSCs exhibited increased expression levels of osteocalcin, peroxisome proliferator-activated receptor gamma, and aggrecan compared to ESK-MSCs. Also, ESK-MSCs exhibited increased expression levels of collagen type I, II, and lipoprotein lipase compared to NM-MSCs. We suggest that the nasal mucosa of bovine could be used as a source of bovine MSCs.

Rabies is a zoonotic disease that causes severe destruction to the central nerve system which is usually fatal. It is one of the most important disease around the world and particular in Asia because of the high costs of prevention and post-exposure treatment. After the recurrence of sylvatic rabies in 1993, the number of raccoon dog mediating rabies cases in Korea has maintained annually until 2011. To better understand the current rabies epidemics in Korea, Korean rabies isolate (SKRBV0601GY) from Gyeonggi province in 2006 was compared with previous isolates in Korea and with isolates originating from the North-East Asia, such as Japan, China and Russia, based on complete nucleoprotein (N) gene sequences. By comparison of the N genes among these viruses, SKRBV0601GY revealed that nucleotide similarity ranged from 97.7 to 99.7%, 96.4 to 97.5%, 91.4 to 96.3%, 89.2 to 90% and 86.1 to 88.1% with Korean isolates, "Arctic-like-2" viruses, "Arctic" viruses, Russian group C - E and Chinese isolates, respectively. Phylogenetic analyses of the isolates revealed that the Korean isolate in 2006 belonged to Korean group B. The topology of the phylogenetic tree of Korean isolates related not the species and year of isolation but the geological location of the virus isolates. All of the Korean isolates showed close relationship to the "Arctic-like-2" virus (Russian group B) more than the "Arctic" virus (Russian group A) and all of the Chinese isolates (Chinese group A, B and C). The "Arctic-like-2" virus group contains the Japanese isolate and Russian group B viruses, originating from the south of East Siberia and Far East in Russia. These molecular data demonstrated that the current rabies epizootic in Korea developed independently of Chinese groups and originated from the "arctic-like-2" viruses in detail.

This study was performed to estimate the seroprevalence of PRRSV in breeding farms in Jeju 2008 using a commercial enzyme-linked immunosorbent assay (ELISA) kit. The tested sera were randomly collected from a total of 1,947 sera from 9 breeding farms unvaccinated in Jeju. As a result, all breeding farms were seropositive for PRRSV. Seven hundred-eighty six of 1,947 sera (40.4%) were positive for PRRSV. Seropositve rate of PRRSV infection in 9 farms showed various levels: 1%, 8.9%, 9.1%, 43%, 46.9%, 48.2%, 51.6%, 60.9%, 85.5%, respectively. The results confirmed that PRRSV infection has been prevailing in breeding farms in Jeju. Also, these results must be taken into a consideration in strategy establishment for the control and eradication of PRRS.