• 한국동물위생학회지
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• 제34권1호
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• pp.13-18
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• 2011

The purpose of this study was to apply a nested-polymerase chain reaction (nPCR) assay to detect and differentiate PCV 2a and PCV 2b. The compared with nPCR and one-step PCR and nPCR showed more sensitive in the detection of PCV-2 from tissue and blood samples. The total of 52 tissue samples was collected from postweanning pigs from 2006 to 2010. All tissue samples showed positive for PCV-2 in one-step PCR and nPCR, followed by the nPCR in order to identify the genotypes of PCV-2. 2 samples (3.8%) showed positive for PCV 2a, and 35 samples were positive for PCV 2b (67.3%), 15 samples (28.9%) were positive the dual genotypes. In addition, 42 blood samples which were collected from the 5 different swine farms were compared figure out the detection rates of nPCR and one-step PCR. The PCV 2 was positive by one-step PCR in 21 samples (50.0%) and nPCR was positive in 37 samples (88.1%). The PCV 2 genotypes in blood samples and 32 samples (76.2%) were positive for PCV 2b and none were positive for PCV 2a, 5 samples (11.9%) were positive for dual genotypes. These results suggest that the nPCR is very efficient for genotyping blood samples and differentiating the genotypes of PCV-2 from field samples.

• International Journal of Industrial Entomology and Biomaterials
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• 제24권1호
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• pp.23-27
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• 2012

Porcine circovirus type 2 (PCV2) is a single-stranded circular DNA virus associated with Postweaning multisystemic wasting syndrome (PMWS), which is considered to be an important infectious swine viral disease. PCV2 capsid protein encoded by ORF2 is a structural protein and expected as the high immunogenicity protein. In this study, we generated recombinant baculovirus containing ORF2 of PCV2 and analyzed the optimal conditions for the production of capsid protein in insect cell. Production and status of recombinant capsid protein in insect cell were confirmed by SDS-PAGE and Western blot analysis using His tag antibody and anti-PCV2 serum. The yield of recombinant capsid protein was high like as shown visible on SDS-PAGE. Optimal multiplicity of infection (MOI) and infection time of recombinant virus were determined as 5 MOI and 4 days, respectively. ORF2 is known to have N-linked glycosylation site, but we couldn't detect the glycosylation of recombinant protein in insect cells.

• 한국동물위생학회지
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• 제45권2호
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• pp.87-99
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• 2022

A novel porcine circovirus 4 (PCV4) was recently emerged in Chinese and Korean pig herds, which provided epidemiological situation where three pathogenic PCVs, PCV2, PCV3, and newly emerged PCV4, could co-infect pig herds in these countries. In this study, a new triplex quantitative real-time polymerase chain reaction (tqPCR) method was developed for the rapid and differential detection of these viruses. The assay specifically amplified each viral capsid gene, whereas no other porcine pathogenic genes were detected. The detection limit of the assay was below 10 copies/µL and the assay showed high repeatability and reproducibility. In the clinical evaluation using 1476 clinical samples from 198 Korean pig farms, the detection rates of PCV2, PCV3 and PCV4 by the tqPCR assay were 13.8%, 25.4%, and 3.8%, respectively, which were 100% agreement with those of previously reported monoplex qPCR assays for PCV2, PCV3, and PCV4, with a κ value (95% CI) of 1 (1.00~1.00). The prevalence of PCV2, PCV3, and PCV4 at the farm levels were 46.5%, 63.6%, and 19.7%, respectively. The co-infection analysis for tested pig farms showed that single infection rates for PCV2, PCV3, and PCV4 were 28.8%, 44.4%, and 9.6%, respectively, the dual infection rates of PCV2 and PCV3, PCV2 and PCV4, and PCV3 and PCV4 were 12.6%, 3.5%, and 5.1%, respectively, and the triple infection rate for PCV2, PCV3, and PCV4 was 1.5%. These results demonstrate that three pathogenic PCVs are widely spread, and their co-infections are common in Korean pig herds, and the newly developed tqPCR assay will be useful for etiological and epidemiological studies of these pathogenic PCVs.

• 한국동물위생학회지
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• 제30권1호
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• pp.85-93
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• 2007

The purpose of this study was to investigate the infection situation of several diseases (post-weaning atrophic pigs) such as porcine reproductive and respiratory syndrome (PRRS) in swine breeding complex in Jeonbuk-Iksan. From February to October in 2006, a total of 28 swine samples (6-10 week old) were collected from 6 farms and examined by polymerase chain reaction(PCR) and clinical signs. In the rate of single infection, pneumonia was top (32.1%), followed by salmonellosis (14.2%)and Glasser's disease (10.7%) and double infection pneumonia/Glasser's disease (17.8%) was detected. PCR was detected of PCV 2 from 28 (100.0%) and PPV 6 (21.4%), PRRS PORF6 10 (35.7%) and POR7 11 (39.2%), but HC and AD was not detected. The results suggest that PCV 2 is complex infection PRRS, PPV and bacterial disease.

• Journal of Veterinary Science
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• 제21권5호
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• pp.78.1-78.15
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• 2020

Background: Enteritis is one of the most frequently reported symptoms in piglets infected with porcine circovirus type 2 (PCV2), but the immunopathogenesis has not been reported. Objectives: This study examined the effect of a PCV2 infection on the intestinal mucosal immune function through morphological observations and immune-related molecular detection. Methods: Morphological changes within the ileum of piglets during a PCV2 infection were observed. The expression of the related-molecules was analyzed using a gene chip. The immunocyte subsets were analyzed by flow cytometry. The secretory immunoglobulin A (SIgA) content was analyzed by enzyme-linked immunosorbent assay. Results: The PCV2 infection caused ileal villus damage, intestinal epithelial cells exfoliation, and an increase in lymphocytes in the lamina propria at 21 days post-infection. Differentially expressed genes occurred in the defense response, inflammatory response, and the complement and coagulation cascade reactions. Most of them were downregulated significantly at the induction site and upregulated at the effector site. The genes associated with SIgA production were downregulated significantly at the induction site. In contrast, the expression of the Toll-like receptor-related genes was upregulated significantly at the effector site. The frequencies of dendritic cells, B cells, and CD8⁺T cells were upregulated at the 2 sites. The SIgA content decreased significantly in the ileal mucosa. Conclusions: PCV2 infections can cause damage to the ileum that is associated with changes in immune-related gene expression, immune-related cell subsets, and SIgA production. These findings elucidated the molecular changes in the ileum after a PCV2 infection from the perspective of intestinal mucosal immunity, which provides insights into a further study for PCV2-induced enteritis.

• 대한수의학회지
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• 제39권2호
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• pp.353-358
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• 1999

New emerging post weaning multisystemic wasting syndrome (PMWS) threatening swine industry worldwide and investigation of the etiological agent is underway. The porcine circovirus (PCV) consistently identified from PMWS pigs and research result indicate that there is strong relationship between PCV and PMWS. Fauns with PMWS submitted pigs suffered from various PMWS typical signs and necropsy finding showed lymph node anomalies. The PCV DNA was amplified from inguinal lymph node collected from pigs with PMWS. PCV specific primers were successfully amplified PCV DNAs and were able to differentiate PCV type I and II. We have identified noble PCV virus with genetic variation. The virus showed insertion of the nucleic acid at the 5' of the genome but did not have PCR product with primer set corresponding to PCV type II virus.

• Journal of Microbiology and Biotechnology
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• 제29권3호
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• pp.482-488
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• 2019

Porcine circovirus type 2 (PCV2) is the causative agent of postweaning multisystemic wasting syndrome (PMWS) in pigs. Replicase (Rep) proteins are considered essential for viral replication. Capsid (Cap) protein is the primary immunogenic protein that induces protective immunity. Little is known about comparison on the immunogenicity of PCV2 Rep and Cap fusion protein and Cap protein. In the present study, recombinant baculoviruses expressing the Rep-Cap fusion protein (Bac-Rep-Cap) and the Cap protein (Bac-Cap) of PCV2 were constructed and confirmed with western blot and indirect fluorescence assay. Immunogenicities of the two recombinant proteins were tested in mice. The titers of antibodies were determined with a PCV2-specific enzyme-linked immunosorbent assay (ELISA) and a serum neutralization assay. The $IFN-{\gamma}$ response of immunized mice was measured by ELISA. The mice immunized with the Bac-Rep-Cap and Bac-Cap successfully produced Cap-specific immunoreaction. The mice immunized with the Bac-Cap developed higher PCV2-specific neutralizing antibody titers than mice injected with the Bac-Rep-Cap. $IFN-{\gamma}$ in the Bac-Rep-Cap group was increased compared to those in the Bac-Cap group. Vaccination of mice with the Bac-Rep-Cap showed significantly decreased protective efficacy compared to the Bac-Cap. Our findings will indubitably not only lead to a better understanding of the immunogenicity of PCV2, but also improved vaccines.

• 대한수의학회지
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• 제44권2호
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• pp.279-286
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• 2004

Total 160 head of porcine kidneys were examined for gross and histopathological lesions and polymerase chain reaction (PCR) for porcine circovirus type 2 (PCV-2), porcine parvovirus (PPV), Leptospira species and porcine reproductive and respiratory syndrome virus (PRRSV). Grossly, 137 kidneys (85.6%) had lesions characterized by the presence of the scattered white foci. Microscopically, multifocal interstitial nephritis, which classified into 4 grades such as, no lesion (Score 0), mild lesion (Score 1), moderate lesion (Score 2) and severe chronic lesion (Score 3) with fibrosis, was observed in 159 cases (99.4%). The histopathologic mean score for multifocal interstitial nephritis was significantly different (P<0.05) between the cases of PCV-2 single infection and the cases of co-infection with PCV-2 and PPV. According to PCR evaluation, PCV-2 were detected in 73.8% (118 cases), PPV were in 66.9% (107 cases), however Leptospira spp. and PRRSV were negative in all kidneys. Both PCV-2 and PPV were detected in 52.5% (84 cases). In 84 cases co-infected with PCV-2 and PPV, the occurrence of lymphoid follicle and vasculitis were observed as 65.5% (55 cases) and 26.2% (22 cases), respectively. These results revealed that PCV-2 and PPV were major infectious agents for interstitial nephritis in slaughtered pigs, Jeju. And the histopathologic lesions of multifocal interstitial nephritis were more severe in the case co-infected with PCV-2 and PPV.

• Journal of Veterinary Science
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• 제21권4호
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• pp.61.1-61.16
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• 2020

Background: Panax notoginseng saponins (PNS) are bioactive substances extracted from P. notoginseng that are widely used to treat cardiovascular and cerebrovascular diseases and interstitial diseases. PNS have the functions of scavenging free radicals, anti-inflammation, improving blood supply for tissue and so on. Objectives: The aim of this study was to investigate the effects of PNS on the oxidative stress of immune cells induced by porcine circovirus 2 (PCV2) infection in vitro and in vivo. Methods: Using an oxidative stress model of PCV2 infection in a porcine lung cell line (3D4/2 cells) and mice, the levels of nitric oxide (NO), reactive oxygen species (ROS), total glutathione (T-GSH), reduced glutathione (GSH), and oxidized glutathione (GSSG) and the activities of xanthine oxidase (XOD), myeloperoxidase (MPO) and inducible nitric oxide synthetase (iNOS) were determined to evaluate the regulatory effects of PNS on oxidative stress. Results: PNS treatment significantly reduced the levels of NO and ROS, the content of GSSG and the activities of XOD, MPO, and iNOS (p < 0.05), while significantly increasing GSH and the ratio of GSH/GSSG in infected 3D4/2 cells (p < 0.05).Similarly, in the in vivo study, PNS treatment significantly decreased the level of ROS in spleen lymphocytes of infected mice (p < 0.05), increased the levels of GSH and T-GSH (p < 0.05), significantly decreased the GSSG level (p < 0.05), and decreased the activities of XOD, MPO, and iNOS. Conclusions: PNS could regulate the oxidative stress of immune cells induced by PCV2 infection in vitro and in vivo.

• 대한수의학회지
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• 제63권1호
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• pp.4.1-4.5
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• 2023

The prevalence of some swine reproductive and zoonotic diseases in Colombia is unknown, making their management difficult. This study assessed the prevalence of porcine circovirus type 3 (PCV3), Leptospira interrogans, pseudorabies virus, and porcine gamma-herpesvirus by polymerase chain reaction in sows in the productive stage, from farms with a history of reproductive failures, at the department of Tolima. The prevalence of PCV3 was 2.6% and 12.6% for L. interrogans, with some samples co-infected with PCV2. Owing to the coinfections with PCV2, it is necessary to establish whether the interactions between these pathogens were related to the presence of reproductive problems.