• 한국농공학회논문집
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• 제54권1호
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• pp.63-72
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• 2012

In livestock industry, damage caused by the epidemic diseases such as Foot-and-Mouth Disease (FMD), Highly-Pathogenic-Avian-Influenza (HPAI) and Porcine-Reproductive-and-Respiratory-Syndrome (PRRS) was very serious. The financial loss incurred from FMD alone which occurred on Nov. 2011 in Korea was estimated at 3 billion won, 23 % of annual livestock industry production. The livestock industry in Korea has greater risk of disease infection because of high density production, etc. Investigating the spread of livestock diseases should consider both direct and indirect contact as well as other various factors including airborne. Airborne infection of livestock disease was first hypothesised in the early 1900s, however, field experimental studies are still limited. Furthermore, no protocol is available in detecting airborne viruses in the field. In this study, effective virus samplers were investigated by comparative analysis of the type of samplers used detect to airborne virus. Laboratory experiments were conducted to compare virus samplers such as Bio-sampler, Dust-sampler, Compact-Cascade-Impactor (CCI) and Microflow in detecting PRRSV. Samples were analyzed by Reverse-Transcription PCR to assess the efficiency of the instrument in detecting the airborne virus. First, samples were classified into five levels according to light intensity of gel images and then the classified results were normalized. In every case, Bio-sampler and Dust-sampler were comparable with each other and have shown to be more effective than CCI and Microflow samplers.

• 한국동물위생학회지
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• 제37권3호
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• pp.143-150
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• 2014

Porcine reproductive and respiratory syndrome virus (PRRSV) is the etiological agent of PRRS characterized by reproductive losses in sows and respiratory disorders in piglets. The PRRSV is a small enveloped virus containing a positive-sense, single-stranded RNA genome and divided into two genotype, type 1 (European) and type 2 (North American), respectively, by nucleotide identity. In this study, ORF7 gene of the type 1 and type 2 PRRSV was cloned and expressed in Baculovirus expression system. Also, monoclonal antibodies (MAbs) against ORF7 were produced and characterized. The expressed ORF7 proteins in the recombinant virus were confirmed by indirect fluorescence antibody (IFA) test using His6 and PRRSV-specific antiserum. A total of eight MAbs were produced and characterized. One (3G12) MAb was type 1 PRRSV ORF7-specific and two (6B10 and 16H8) were type 2 PRRSV ORF7-specific. Other five (1A1, 2A4, 4B4, 12C4 and 13F11) MAbs reacted with both type 1 and type 2 PRRSV. Some PRRSV ORF7-specific MAbs recognized the porcine tissues infected with PRRSV by IFA or immunohistochemistry (IHC) assay. From this experiment, it was confirmed that MAbs produced in this study were PRRSV ORF7-specific and could be used as reliable reagents for type 1/type 2 PRRSV detection.

• Journal of Microbiology and Biotechnology
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• 제31권7호
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• pp.942-948
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• 2021

Canine influenza virus (CIV) induces acute respiratory disease in dogs. In this study, we aimed to determine the signaling pathways leading to the induction of IFN-β in a canine respiratory epithelial cell line (KU-CBE) infected with the H3N2 subtype of CIV. Small interfering RNAs (siRNAs) specific to pattern recognition receptors (PRRs) and transcription factors were used to block the IFN-β induction signals in H3N2 CIV-infected KU-CBE cells. Among the PRRs, only the TLR3 and RIG-I expression levels significantly (p < 0.001) increased in CIV-infected cells. Following transfection with siRNA specific to TLR3 (siTLR3) or RIG-I (siRIG-I), the mRNA expression levels of IFN-β significantly (p < 0.001) decreased, and the protein expression of IFN-β also decreased in infected cells. In addition, co-transfection with both siTLR3 and siRIG-I significantly reduced IRF3 (p < 0.001) and IFN-β (p < 0.001) mRNA levels. Moreover, the protein concentration of IFN-β was significantly (p < 0.01) lower in cells co-transfected with both siTLR3 and siRIG-I than in cells transfected with either siTLR3 or siRIG-I alone. Also, the antiviral protein MX1 was only expressed in KU-CBE cells infected with CIV or treated with IFN-β or IFN-α. Thus, we speculate that IFN-β further induces MX1 expression, which might suppress CIV replication. Taken together, these data indicate that TLR3 and RIG-I synergistically induce IFN-β expression via the activation of IRF3, and the produced IFN-β further induces the production of MX1, which would suppress CIV replication in CIV-infected cells.

• International Journal of Industrial Entomology and Biomaterials
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• 제32권2호
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• pp.90-97
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• 2016

The major structural proteins of porcine reproductive and respiratory syndrome virus (PRRSV) are derived from ORFs 4, 5, and 6. They have been considered very important to arouse the humoral and cellular immune responses against PRRSV infection and proposed to be the excellent candidate proteins in the design of PRRS bioengineering vaccine. However, the PRRSV structural proteins are produced in low levels in the infected cells because it forms insoluble protein and possesses several transmembrane regions. To overcome this problem, we fused the ORF4, ORF5, and ORF6 with SUMO (small ubiquitin-related modifier). The resulting fusion protein SUMO-ORF4, -ORF5, and -ORF6 were highly expressed in Bm5 cells. The level of protein expression using the Bombyx mori larvae was higher than that using Bm5 cells. In addition, fusion to SUMOstar, which is not processed by native SUMO proteases, significantly enhanced protein expression levels compared to SUMO fusion. This study demonstrated that SUMO or SUMOstar, when fused with PRRSV structural proteins, was able to promote its soluble expression. This may be a better method to produce PRRSV structural proteins for vaccine development.

• 한국동물위생학회지
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• 제46권1호
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• pp.59-66
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• 2023

Wild boar is closely related to domestic pigs in terms of genetic homogeneity and the possibility of a source of infection by contact. This study investigated the prevalence of viral diseases from wild boars inhabiting Gyeongsangnam-do, South Korea. A total of 374 blood samples were collected and subjected to antigen tests to detect African swine fever virus (ASFV), Porcine circovirus type-2 (PCV2), Porcine reproductive and respiratory syndrome virus (PRRSV). For seroprevalence, PCV2, PRRS, classical swine fever virus (CSFV), Aujezsky's disease (ADV), and foot and mouth disease virus (FMDV) were investigated. The antigenic analysis revealed 73 positive cases (19.5%) for PCV2, while no positive cases for ASFV and PRRSV. For the antibody test, 225 (60.2%), 2 (0.5%), and 48 (12.8%) cases were detected against PCV2, PRRSV, and CSFV, respectively. There were no antibodies detected against both ADV and FMDV. Our results suggest that the viruses infecting both wild boar and domestic pig, mainly PCV2, are circulating in the wild boar population thus, the consistent monitoring of prevalence in wild boar will be needed for transboundary spillover to the domestic pig.

• 대한수의학회지
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• 제43권3호
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• pp.463-469
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• 2003

Eight nursery to grower pigs exhibiting weight loss and sudden death were diagnosed as postweaning multisystemic wasting syndrome (PMWS) based on the results of gross findings, histopathology, immunohistochemistry, fluorescent antibody test, virus isolation, PCR, serology, and electron microscopy. Groosly, the pigs had a rough hair coats and were severely emaciated. And moot lymph nodes were pale and enlarged. Lungs were not fully collapsed and exhibited 10 to 40% pale red cranioventral consolidation. Histopathologically, typical lymphohistiocytic interstitial to bronchointerstitial pneumonia, chronic lymphadenitis, severe lymphoid depletion, and basophilic intracytoplasmic inclusions were noted in the most lymphoid tissues. Porcine circovirus panicles were observed in the inguinal lymph node of the pigs by electron microscopy. Porcine circovirus type 2 (PCV2) antigens or viral DNAs were detected in the lesions of all pigs using immunohistochemistry or PCR. Two PCV2 were isolated from a homogenate of pooled lung and lymph node in 2 of the 5 pigs. Additionally, antigens of porcine reproductive and respiratory syndrome virus (PRRSV) and Hemophilus (H.) parasuis were also detected by immunofluorescent antibody test. Serologically, 55% of randomly selected sows and fattening pigs was serum antibody positive to PCV2 by an indirect fluorescent antibody (IFA) test and approximately 18 % of animals in the herd were serologically pooitive by the ELISA kit for PRRSV. To our knowledge, this is the first report of PMWS co-infected with PCV-2, PRRS, and H. parasuis in Korea.

• 한국동물위생학회지
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• 제24권4호
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• pp.359-367
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• 2001

A serological survey was performed to establish basic data for the prevalence of antibodies to some major diseases of domesticated boar serum samples from January to December 2000. Sera collected in breeding farms in Gyeongbuk province were tested for Aujeszky's disease virus(ADV), Porcine reproductive and respiratory syndrome virus(PRRSV), Porcine parvovirus(PPV), Japanese encephalitis virus (JEV), Bordetella bronchiseptica(B bronchiseptica), Mycoplasma ; APP), Toxoplasma, and Brucella. There was no antibody to ADV in domesticated boars serum samples detected by Anti-ADV-gpI assay kit. Sero-positive samples to PRRS by IFA were 0.9%(3/330) The HI titers to PPV ranged variously from less than 10 to over 1,280. Two hundred ninety-four out of 330 tested sera showed HI titer of less than 10. In HI test to JEV, 90.3% of the sera (298/330) were below 10. The majority of the serum samples had low prevalence of the antibody B bronchiseptica. ELISA titers to M hyopneumoniae ranged variously from $\leq$ 10 to $\geq$ 1,280. Antibody titers to A pleuropneumoniae type 2(APP2) and type 5(APP5) were 58.2% and 52.7%, respectively, and the tested samples showing ELISA antibody titers of less than 20. There was no significant geographical difference between APP2 and APP5 in this study. In the antibody test of Toxoplasma, 11.5%(38/330) were positive and samples were all negative in sera test of Brucella.

• 대한수의학회지
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• 제34권1호
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• pp.77-83
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• 1994

유사산 태아의 폐, 청색증을 나타내는 자돈으로부터 돼지생식기 및 호흡기증후군(PRRS)의 원인체로 추정되는 바이러스주(KPRRSV) 들을 분리하였다. 분리된 바이러스주는 돼지콜레라, 돼지오제스키병, 돼지뇌심근염바이러스에 대한 형광항체반응에서는 음성이었으며 기니픽혈구에 대한 혈구응집 능력을 나타내지 않았다. 그리고 포유 마우스의 뇌내 접종시 이상을 나타내지 않았으나 돼지생식기 및 호흡기증후군에 대한 형광항체검사시 양성반응을 나타내었다. 분리된 바이러스는 돼지폐포탐식세포(porcine alveola macrophages)에서 세포변성효과(cytopathic effect)를 나타내었으며 세포변성효과를 나타내었던 바이러스주중 일주(KPRRSV-1)를 돼지폐포탐식세포에서 7대 연속 계대하여 돼지에 접종한 후 혈청을 분리하여 미국 및 유럽지역에서 분리된 돼지유행성 유사산 및 호흡기증후군의 바이러스를 탐식세포에 감염시켜 효소면역방법 (immunoperoxidase monolayer assay)으로 분석한 결과 분리된 바이러스는 미국형 돼지호흡기 및 유사산증후군에 가까운 항원형으로서 판명되었다.

• 한국동물위생학회지
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• 제26권2호
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• pp.163-184
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• 2003

This study was conducted to survey the farm which suffered from disease similar to classical swine fever(CSF) in Gyeongbuk province. Clinical signs appeared first in a few number of growing pigs which showed specific signs of diarrhea, depression, sleepiness, and reluctance to get up or to eat. Younger piglets may have appeared chilled, shiver and huddle together. As the disease progresses the affected pig's skin went red and purple. In histopathological signs, there were many haemorrhages throughout the body and larger haemorrhages in some organs such as lymph nodes. And there is a precipitous fall in the number of circulating leukocytes in the blood. In spite of insisting of farmer which did not vaccinate to classical swine fever, significant antibody production was detected in these affected pigs at enzyme-linked immuonsorbent assay. According to the above results at first glance, these affected pig suspected with CSF in clinical signs and histopathological lesions only. Because the symptoms and post-mortem picture were very similar to CSF, these false positive results would have been dangerous to diagnostician. But by reverse transcriptase polymerase chain reaction(RT-PCR) and comparative nucleotide sequence analysis, the disease was correctly diagnosed with post-weaning multisystemic wasting syndrome(PMWS) and porcine reproductive and respiratory syndrome(PRRS) compoundly. And the antigen which were detected the lesion similar to CSF virus, was confirmed with LOM vaccine strain of CSF. In most national CSF eradication program and in countries which are free of the CSF virus, vaccination against CSF is not practiced and generally is not allowed. But now in Korea, routine vaccination is practiced because of outbreaking the CSF repeatedly. When CSF is diagnosed the whole herd and other in contact animal are slaughtered continuously.

• 대한수의학회지
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• 제37권4호
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• pp.793-807
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• 1997

We tried to develop detection system of porcine reproductive and respiratory syndrome virus(PRRSV) by in situ hybridization(ISH) in the piglets experimentally infected with KPRRS-2, the Korean isolate(12 piglets) or Mn-1b, the American isolate(4 piglets), and in the natural infection suspected 6 piglets. Twelve 30-days-old piglets(two pigs per each inoculated group) were inoculated by nasal instillation of KPRRS-2 virus(total dose $10^{4.5}TCID_{50}$), Six piglets(one pig per each group) were induced contact infection with inoculated piglets, during the experiment, and two piglets were used as control. Inoculated or contacted piglets were euthanized at 1, 3, 5, 7, 14 and 21 days postinoculation(DPI). The respiratory signs such as coughing and nasal discharge were observed on day 3 DPI, and ear cyanosis were on day 5 DPI, including contacted piglets. Through the necropsy, purple discolorization of dorsal part of lung, and hypertrophy of local lymph nodes were observed. The histopathological lesions of lung were interstitial pneumonia characterized by type 2 pneumocyte hyperplasia. We prepared the probe for ISH by RNA isolation from KPRRS-2, RT-PCR, and biotin labeling. We performed the ISH within only 1~2 hours using $Microprobe^{TM}$ capillary action system. As the results, the strong red specific positive signals, means PRRSV distribution, was mainly observed in the cytoplasm of alveolar macrophages. And also signals were detected in some type 2 pneumocytes and bronchiolar epithelium of lung, myocardium, liver, kidney, tonsil, spleen, gastrointestinal mucosa, testis and lymph nodes.