Mesenchymal stem cells (MSCs) possess the ability to differentiate into multiple cell lineages, and thus, confer great potential for use in regenerative medicine and biotechnology. In the present study, we attempted to isolate and characterize bovine tongue tissue epithelium-derived MSCs (boT-MSCs) and investigate the culture conditions required for long-term culturing of boT-MSCs. boT-MSCs were successfully isolated by the collagenase digestion method and their proliferative capacity was maintained for up to 20 or more passages. We observed a significant increase in the proliferation of boT-MSCs during the 20 consecutive passages under low-glucose Dulbecco's modified Eagle's medium culture condition among the three culture conditions. These boT-MSCs presented pluripotency markers (octamer-binding transcription factor 3/4 (Oct3/4) and sex determining region Y-box2 (Sox2)) and cell surface markers, which included CD13, CD29, CD44, CD73, CD90, CD105, CD166, and major histocompatibility complex (MHC) class I (MHC-I) but not CD11b, CD14, CD31, CD34, CD45, CD80, CD86, CD106, CD117, and MHC-II at third passage. Moreover, these boT-MSCs could differentiate into mesodermal (adipocyte, osteocyte, and chondrocyte) cell lineages. Thus, the present study suggests that the tongue of bovines could be used as a source of bovine MSCs.

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.

Natural killer (NK) cells play have a crucial role in the early phase of immune responses against various pathogens. We compared characteristics of canine NK cells against two canine mammary carcinoma cell lines, REM134 and CF41.Mg. REM134 showed higher expression of progesterone receptor, proliferative cell nuclear antigen, Ki67, multiple drug resistance, Bmi-1, c-myc, E-cadherin, and human epidermal growth factor receptor type-2 than that of CF41.Mg. For specific expansion and activation of NK cells, we isolated CD5 negative cells from canine peripheral blood mononuclear cells and co-cultured K562 cells in the presence of interleukin (IL)-2, IL-15, and IL-21 for 21 days. As a result, we found that expression markers of activated NK cells such as NKp30, NKp44, NKp46, NKG2D, CD244, perforin, granzyme B, and tumor necrosis factor alpha were highly upregulated. In addition, we found there was upregulated production of interferon gamma of activated NK cells against target cells such as REM134 and CF41.Mg. Specifically, we observed that cytotoxicity of NK cells against target cells was more sensitively reacted to CF41.Mg than REM134. Based on the results of this study, we recommend the development of an experimental application of CF41Mg, which has not been reported in canine mammary carcinoma research. Keywords: canine, mammary carcinoma, natural killer cells, interferon gamma, cytotoxicity

A maximum clade credibility tree constructed using the full-length spike (S) and hemagglutinin-esterase genes revealed that Vietnamese Bovine coronavirus (BCoV) strains belong to a single cluster (C1); therefore, they might share a common origin with Cuban and Chinese BCoV strains. The omega values of cluster 1 (C1) and cluster 2 (C2) were 0.15734 and 0.11613, respectively, and naive empirical bayes analysis identified two amino acid positions (179 and 501) in the S protein in C1 and three amino acid positions (113, 501, and 525) in that of C2 that underwent positive selection (p > 99%). The evolutionary rate of C1 was estimated to be 7.6206 × 10−4 substitutions/site/year, and the most recent common ancestor (tMRCA) of Vietnamese BCoVs was estimated to date back to 1962 (95% HPD 1950–1973). The effective population sizes of C1 and C2 underwent a rapid reduction after 2000 and 2004, respectively.

Here, we examine the effects of LOM(Low virulence of Miyagi) strains isolated from pigs (Jeju LOM strains) of Jeju Island, where vaccination with a live attenuated classical swine fever (CSF) LOM vaccine strain was stopped. The circulation of the Jeju LOM strains was mainly caused by a commercial swine erysipelas (Erysipelothrix rhusiopathiae) vaccine mixed with a LOM vaccine strain, which was inoculated into pregnant sows of 20 pig farms in 2014. The Jeju LOM strain was transmitted to 91 pig farms from 2015 to 2018. A histopathogenic investigation was performed for 25 farms among 111 farms affected by the Jeju LOM strain and revealed pigs infected with the Jeju LOM strain in combination with other pathogens, which resulted in the abortion of fetuses and mortality in suckling piglets. Histopathologic examination and immunohistochemical staining identified CSF-like lesions. Our results also confirm that the main transmission factor for the Jeju LOM strain circulation is the vehicles entering/exiting farms and slaughterhouses. Probability estimates of transmission between cohabiting pigs and pigs harboring the Jeju LOM strain JJ16LOM-YJK08 revealed that immunocompromised pigs showed horizontal transmission (r = 1.22). In a full genome analysis, we did not find genetic mutation on the site that is known to relate to pathogenicity between Jeju LOM strains (2014–2018) and the commercial LOM vaccine strain. However, we were not able to determine whether the Jeju LOM strain (2014–2018) is genetically the same virus as those of the commercial LOM vaccine due to several genetic variations in structure and non-structure proteins. Therefore, further studies are needed to evaluate the pathogenicity of the Jeju LOM strain in pregnant sow and SPF pigs and to clarify the characteristics of Jeju LOM and commercial LOM vaccine strains.

Classical swine fever virus (CSFV) strains ND20 and HY78 were detected from infected pigs in the Xuan Truong-Nam Dinh and Hung Yen provinces in North Vietnam in 2014 and 2015, respectively. The most prevalent CSFV subgenotypes in Vietnam are 2.1 and 2.2, and these two complete genome sequences will help the CSFV prevention policy in Vietnam. In particular, subgenotype 2.2 (ND20 strain) has been reported less worldwide, so it is worth sharing information about this subgenotype.

Here, we constructed an attenuated live marker classical swine fever (CSF) vaccine (Flc-LOM-BErns) to eradicate CSF. This was done by taking infectious clone Flc-LOM, which is based on an attenuated live CSF vaccine virus (LOM strain), and removing the full-length classical swine fever virus (CSFV) E-rns sequences and the 3' end (52 base pairs) of the CSFV capsid. These regions were substituted with the full-length bovine viral diarrhoea virus (BVDV) E-rns gene sequence and the 3' end (52 base pairs) of the BVDV capsid gene. Sows were vaccinated with the Flc-LOM-BErns vaccine 3 weeks before insemination and then challenged with virulent CSFV at the early, mid- or late stages of pregnancy. We then examined transplacental transmission to the foetuses. Piglets born to sows vaccinated with Flc-LOM-BErns did not show vertical infection, regardless of challenge time. In addition, CSFV challenge did not affect the delivery date, weight or length of the foetus. Pregnant sows inoculated with the Flc-LOM-BErns vaccine were anti-CSF E-rns antibody-negative and anti-BVDV E-rns antibody-positive. Challenge of pregnant sows with virulent CSFV resulted in anti-CSF E-rns antibody positivity. These results strongly indicate that differential diagnosis can be conducted between the Flc-LOM-BErns vaccinated animal and virulent CSFV affected animal by detecting antibody against BVDV E-rns or CSF E-rns gene. Therefore, the Flc-LOM-BErns vaccine may fulfil the function of differential diagnosis which required for DIVA vaccine. (C) 2019 Elsevier Ltd. All rights reserved.

A preliminary study into the protective mechanisms of adaptive immunity against porcine reproductive and respiratory syndrome virus (PRRSV) in piglets (n = 9) born to a gilt challenged intranasally with a type-2 PRRSV.Immune parameters (neutralizing antibodies, CD3 + CD4 + , CD3 + CD8 + , CD3 + CD4 + CD8 + T-lymphocytes, and PRRSVspecific interferon (IFN)-γ secreting T-lymphocytes) were compared with infection parameters (macro-and microscopic lung lesion, and PRRSV-infected porcine alveolar macrophages (CD172α + PRRSV-N + PAM) as well as with plasma and lymphoid tissue viral loads.Percentages of three T-lymphocyte phenotypes in 14-days post-birth (dpb) peripheral blood mononuclear cell (PBMC) had significant negative correlations with percentages of CD172α + PRRSV-N + PAM (p < 0.05) as well as with macroscopic lung lesion (p < 0.01).Plasma and tissue viral loads had significant (p < 0.05) negative correlations with CD3 + CD4 + CD8 + T-lymphocyte percentage in PBMC.Frequencies of CD3 + CD8 + and CD3 + CD4 + T-lymphocytes in 14-dpb PBMC had significant negative correlations with of lymph node (p = 0.04) and lung (p = 0.002) viral loads.IFN-γ-secreting T-lymphocytes frequency had a significant negative correlation with gross lung lesion severity (p = 0.002).However, neutralizing antibody titers had no significant negative correlation (p > 0.1) with infection parameters.The results indicate that T-lymphocytes contribute to controlling PRRSV replication in young piglets born after in-utero infection.

Porcine reproductive and respiratory syndrome (PRRS) is recognized as one of the most important infectious diseases causing serious economic loss in the swine industry worldwide. Due to its increasing genetic diversity, a rapid and accurate diagnosis is critical for PRRS control. The immunochromatographic strip test (ICST) is a rapid and convenient type of immunoassay. In this study, an on-site immunochromatographic assay-based diagnostic method was developed for detection of PRRS virus (PRRSV)-specific antibodies. The method utilized colloidal gold nanoparticle-labeled dual-type nucleocapsid proteins encoded by open reading frame 7. We evaluated 991 field samples from pig farms and 66 serum samples from experimentally PRRSV-inoculated pigs. Based on true PRRSV-specific antibody-positive or -negative sera determined by immunofluorescence assay and IgM enzyme-linked immunosorbent assay (ELISA), the specificity and sensitivity of the ICST were 97.5% and 91.1%, respectively, similar to those of a commercial ELISA (IDEXX PRRS X3 Ab). More importantly, the ICST was completed within 15 min and could detect the PRRSV-specific antibody at an earlier stage of infection (3-7 days) than that of ELISA (7+ days). The results demonstrate that the developed ICST has great potential as an on-farm diagnostic method, providing excellent diagnostic performance in a quick and convenient manner.

We report here the genome sequence of the influenza A virus strain A/swine/Korea/61/2016, isolated from swine in the Republic of Korea. On the basis of sequence analysis, A/swine/Korea/61/2016 is marked from swine H1N1 influenza virus.

Porcine 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.

Rabies virus (RABV), canine distemper virus (CDV), canine parvovirus type-2 (CPV-2), and canine influenza A virus (CIV) are important contagious pathogens in canine populations. To assess post-vaccination immunity against RABV, CDV and CPV-2, and serological evidence of exposure to influenza A virus in military working dogs (MWDs) in Korea, we tested blood samples of 78 MWDs by fluorescent antibody virus neutralization (FAVN) for RABV, and by commercially available enzyme-linked immunosorbent assay (ELISA) for CDV, CPV-2, and CIV. Korean MWDs had high antibody-positive rates against RABV (97.4%, ≥0.5 IU/ml), CDV (94.8%), and CPV (100%). All dogs tested seronegative (0/78; 0%) for influenza A virus. Two 1-year-old dogs stationed in known rabies outbreak areas (Gangwon and Gyeonggi) exhibited VNA titers below the protective level (0.06 and 0.29 IU/ml, respectively). The breed and sex of MWDs were not significantly associated with antibody titers for RABV, CDV, or CPV; however, age was significantly associated with CPV antibody titers, while region of residence was associated with CDV antibody titer. Taken together, the data presented here provide important insights necessary for post-vaccination management and control of infectious diseases in MWDs.

Endemic animal rabies causes >99% of all human rabies cases; elimination of animal rabies reduces the rate of human infections. The most recent animal rabies cases in the Gangwon and Gyeonggi provinces of Korea occurred in November 2012 and February 2013, respectively. Here we explore ways to ensure that Korea remains animal rabies non-occurrence. The government must prevent rabies recurrence by vaccinating dogs, distributing bait vaccine in regions with a high rabies risk, performing laboratory-based surveillance, preventing the flow of rabies-suspect animals from neighboring countries, and enhancing border quarantine. As has already been shown in several developed countries, careful and ongoing rabies control will allow Korea to remain animal rabies-free.

The 5' UTR (n =102) and full-length E2 (n = 37) genes of classical swine fever viruses (CSFVs) circulating in South Korea over the past 30 years (1987-2017) were examined to determine the evolutionary rate and estimated time of the most recent common ancestor (tMRCA). From 2000, the Korean classical swine fever (CSF) antigen changed from genotype 3 to 2, which comprises subgenotypes 2.1b (2002-2013) and 2.1d (2011-2017). There are genotypic variations in the full-length E2 gene of Korean CSFV genotypes 2.1b and 2.1d (seven separate amino acid substitutions); these are useful distinguishing markers. The mean substitution rate ( x 10(3) substitutions/site/year) for Korean CSFV was estimated to be 2.2088 (95% highest posterior density (HPD): lower, 1.7045; upper, 2.7574) and the mean tMRCA was estimated to be 1901 (95% HPD: lower, 1865; upper, 1933). The effective population size of Korean CSFV genotype 2 increased rapidly from 2002 to 2003, after which it remained constant. The occurrence of CSF in Korea is expected to decline in the future; however, it will likely be more prevalent in wild boar than in domestic pigs. Thus, there is a risk of transmission from wild boar to breeding pigs.