• 대한수의학회지
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• 제39권2호
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• pp.311-317
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• 1999

Pathogenic effects of 29 different porcine reproductive and respiratory syndrome(PRRS) virus isolates were investigated in swine tracheal ring(STR) cultures by examining their effects on the ciliary activity of STR. Inhibition of ciliary movement and destruction of the tracheal epithelium were seen between 72 and 96 hours postinoculation(PI). Virus replication was demonstrated by examining viral infectivity of the supernatants from the STR cultures. PRRS virus antigen in macrophages was detected by a streptavidin-biotin complex(ABC) immunoperoxidase method. Of the 29 PRRS virus isolates, 8 isolates were classified into pathogenic, and the remaining 21 isolates were determined as mildly pathogenic or apathogenic viruses. These results suggest that STR examination may be used as a method for predicting pathogenic variability of PRRS virus isolates.

• 한국수의병리학회:학술대회논문집
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• 한국수의병리학회 2003년도 추계학술대회초록집
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• pp.32-32
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• 2003

Intestinal infections are common in growing pigs and can be caused by multiple pathogens, environmental and management factors [1]. Among the most important viruses in swine enteritis are porcine epidemic diarrhea virus (PEDV), transmissible gastroenteritis virus (TGEV), porcine enteric calicivirus (PECV), porcine group A rotavirus (PRV gp A) and bacteria are Escherichia coli and Salmonella spp. and protozoa is Isospora suis [1]. The DNA chip system can serve as a powerful tool that can be utilized for simultaneous detection of specific pathogenic bacteria strains and viruses [2,3]. The combination of PCR and DNA chip technology will provide a novel method for the detection of porcine enteric pathogens thus revolutionize the diagnosis and management of the disease. The aim of this study is to develop DNA chip system for the rapid and reliable detection of five major porcine enteric pathogens based on oligonucleotide DNA chip hybridization. (omitted)

• 한국수정란이식학회지
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• 제1권1호
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• pp.16-27
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• 1986

Based on the current importing and exporing regulations for disease control of embryo transfer, some important microorganisms and their control possibilities are reviewed. The results reviewed were sumrnarized as follows: 1. Regulations regarding to the import of embryos vary between importing and exporting countries, but exporting countries examine the donor and embryos for the heaith certification by the requirements of importing countries. 2. Organisms that infect the gametes are 5 kinds of viruses and the diseases caused by them could not be controlled or eradicated using embryo transfer. 3. Organisms that do not infect the gametes are 4 kinds of viruses and the causal organisms are potential candidates for control or eradication by embryo transfer. 4. Organisms that penetrate the zona pellucida and infect the embryo are 6 kinds of viruses including bovine viral diarrhea virus. 5. Organisms that cannot penetrate the zona pellucida or do not infect the embryo are 15 kinds of viruses and the removal from their contaminations are recommended by proper washing procedure and antisera treatment. Bovine and porcine parvovirus, porcine pseudorabies virus and vesicular stomatitis virus are included in these organisms. 6. Bovine embryos that artificially exposed to various pathogenic organisms such as bovine herpes virus, IBR virus, bluetongue virus, bovine viral diarrhea virus and Brucella abortus in vitro are discussed about their infection by several treatments.

• 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.

• 대한수의학회지
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• 제39권2호
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• pp.318-326
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• 1999

Respiratory pathogenic effects of several porcine reproductive and respiratory syndrome virus(PRRSV) isolates were examined in swine tracheal ring(STR) cultures by examining their effect on ciliary activity. One high and one low pathogenic PRRSV isolates were then selected and their pathogenicity investigated in 3-week-old conventional PRRSV-seronegative pigs. Ten pigs each were inoculated intranasally with the high or low pathogenic PRRSV isolate and 6 pigs were sham inoculated as negative controls. Two pigs each from the inoculated group and one pig each from negative control group were killed on 4, 7, 14, 21 and 28 days postinoculation(pI). At necropsy, degrees of gross lung lesion was determined. Turbinate, tonsil, trachea and lung samples were collected for virus isolation or histopathology. Gross lung lesions were observed mainly on 14 days PI with high and low pathogenic isolates inducing moderate diffuse and mild gross lung lesions, respectively. Inoculation of either the high or low pathogenic virus resulted in loss of cilia in ciliated epithelium of turbinates and trachea between 7 and 28 days PI. High pathogenic virus caused increased number of Goblet cells in the tracheal epithelial layer between 4 and 21 days PI whereas the low pathogenic virus did it between 14 and 28 days PI and with a lesser degree. Although both viruses produced interstitial pneumonia, the lesion was less severe with the low pathogenic virus. The isolation of high pathogenic virus from tissues and sera was earlier and more consistent than that of the low pathogenic virus. The agreement between in vitro and in vivo tests indicates that STR cultures may be used as a routine method to determine the respiratory pathogenicity of PRRSV isolates.

• 미생물학회지
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• 제52권2호
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• pp.140-147
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• 2016

동물유래성분을 원재료로 사용하는 의료기기는 원료물질 유래 바이러스가 오염될 가능성이 있기 때문에 생산과정 중 바이러스가 오염되지 않도록 하여야 한다. 또한 생산공정은 오염될 가능성이 있는 바이러스들을 불활화하거나 제거하는 공정을 포함하여야 한다. 본 연구를 통해 돼지유래 이종골을 원재료로 사용한 바이러스 안전성이 보증된 치과용 골이식재(THE Graft$^{(R)}$) 제조공정을 개발하였다. THE Graft$^{(R)}$ 제조공정은 30% 과산화수소수와 80% 에탄올을 각각 처리하여 지방을 제거하는 공정과 $1,300^{\circ}C$ 열처리 공정을 통해 콜라겐과 유기물을 제거하는 공정을 포함한다. 또한 최종적으로 생산된 hydroxyapatite 성분의 골이식재에 25 kGy의 감마선을 조사하여 감염성 위해인자를 불활화하는 공정을 포함한다. THE Graft$^{(R)}$의 형태학적 특성을 소유래 hydroxyapatite 성분의 골이식재인 Bio-Oss와 SEM과 TEM을 이용하여 비교한 결과 구조적 특성이 유사함을 확인하였다. $1,300^{\circ}C$ 열처리 공정과 25 kGy 감마선 조사 공정의 바이러스 불활화 효과를 평가하기 위해 transmissible gastroenteritis virus (TGEV), pseudorabies virus (PRV), porcine rotavirus (PRoV), porcine parvovirus (PPV)를 모델 바이러스로 선정하였다. $1,300^{\circ}C$ 열처리 공정에서 TGEV, PRV, PRoV, PPV 모두 검출한계 이하로 불활화되었으며, 바이러스 로그 감소 값은 각각 ${\geq}4.65$, ${\geq}5.81$, ${\geq}6.28$, ${\geq}5.21$이었다. 또한 감마선 조사 공정에서도 TGEV, PRV, PRoV, PPV 모두 검출한계 이하로 불활화되었으며, 바이러스 로그 감소 값은 각각 ${\geq}4.65$, ${\geq}5.87$, ${\geq}6.05$, ${\geq}4.89$이었다. 두 공정에서 TGEV, PRV, PRoV, PPV의 누적 바이러스 로그 감소 값은 각각 ${\geq}9.30$, ${\geq}11.68$, ${\geq}12.33$, ${\geq}10.10$이었다. 이상의 결과에 의하면, THE Graft$^{(R)}$ 제조공정은 바이러스 안전성 보증을 위한 충분한 바이러스 불활화 능력을 가지고 있는 것으로 판단된다.

• Journal of Microbiology and Biotechnology
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• 제10권6호
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• pp.858-864
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• 2000

The purpose of the present study was to examine the efficacy and mechanism of the fraction IV cold ethanol fractionation and pasteurization ($60^{\circ}C$ heat treatment for 10h) steps, involved in the manufacture of albumin from human plasma, in the removal and/or inactivation of blood-born viruses. A variety of experimental model viruses for human pathogenic viruses, including the Bovine viral diarrhoea virus (BVDV), Bovine herpes virus (BHV), Murine encephalomyocarditis virus (EMCV), and Porcine parvovirus (PPV), were selected for this study. Samples from the relevant stages of the production process were spiked with the viruses, and the amount of virus in each fraction was then quantified using a 50% tissue culture infectious dose ($TCID_{50}$). The mechanism of reduction for the enveloped viruses (BHV and BVDV) during fraction IV fractionation was inactivation rather than partitioning, however, it was partitioning in the case of the non-enveloped viruses (EMCV and PPV). The log reduction factors achieved during fraction IV fractionation were ${\geq}6.9$ BHV, $\geq5.2$ for BBDV, 4.9 for EMC, and 4.0 for PPV. Pasteurization was found to be a robust and effective step in inactivating the enveloped viruses as well as EMCV. The log reduction factors achieved during pasteurization were $\geq7.0$ for BHV, $\geq6.1$ for BVDV, $\geq6.3$ for EMCV, and 1.7 for PPV. These results indicate that the production process for albumin has sufficient virus-reducing capacity to achieve a high margin for virus safety.

• Journal of Microbiology and Biotechnology
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• 제14권1호
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• pp.140-147
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• 2004

Urokinase is an enzyme with fibrinolytic activity (plasminogen activator) isolated from fresh urine of healthy men. Viral safety is an important prerequisite for clinical preparation of the protein from urine. In order to increase the viral safety of a high purity urokinase in regard to non-enveloped viruses, a virus removal process using a novel polyvinylidene fluoride membrane filter (Viresolve NFP) has been optimized. Urokinase was able to pass through the filter with recoveries of 95% in the production scale process. No substantial changes were observed in physical and biochemical characteristics of the filtered urokinase in comparison with those of the enzyme before filtration. A 47-mm disk membrane filter was used to simulate the process performance of the production scale cartridges and tested if it could remove several experimental model viruses for human pathogenic viruses, including porcine parvovirus (PPV), human hepatitis A virus (HAV), murine encephalomyocarditis virus (EMCV), bovine viral diarrhoea virus (BVDV), and bovine herpes virus (BHV). Non-enveloped viruses (PPV, HAV, and EMCV) as well as enveloped viruses (BVDV and BHV) were completely removed during filtration. The log reduction factors achieved were $\geq$4.86 for PPV, $\geq$4.60 for HAV, $\geq$6.87 for EMCV, $\geq$4.60 for BVDV, and $\geq$5.44 for BHV. These results indicate that the virus filtration process successfully improved the viral safety of the final products.

• Journal of Microbiology and Biotechnology
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• 제18권7호
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• pp.1317-1325
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• 2008

Viral safety is an important prerequisite for clinical preparations of plasma-derived pharmaceuticals. One potential way to increase the safety of therapeutic biological products is the use of a virus-retentive filter. In order to increase the viral safety of human antihemophilic factor IX, particularly in regard to non-enveloped viruses, a virus removal process using a polyvinylidene fluoride membrane filter (Viresolve NFP) has been optimized. The most critical factor affecting the filtration efficiency was operating pH and the optimum pH was 6 or 7. Flow rate increased with increasing operating pressure and temperature. Recovery yield in the optimized production-scale process was 96%. No substantial changes were observed in the physical and biochemical characteristics of the filtered factor IX in comparison with those before filtration. A 47-mm disk membrane filter was used to simulate the process performance of the production-scale cartridges and to test if it could remove several experimental model viruses for human pathogenic viruses, including human hepatitis A virus (HAV), porcine parvovirus (PPV), murine encephalomyocarditis virus (EMCV), human immunodeficiency virus type 1 (HIV), bovine viral diarrhea virus (BVDV), and bovine herpes virus (BHV). Non-enveloped viruses (HAV, PPV, and EMCV) as well as enveloped viruses (HIV, BVDV, and BHV) were completely removed during filtration. The log reduction factors achieved were $\geq$6.12 for HAV, $\geq$4.28 for PPV, $\geq$5.33 for EMCV, $\geq$5.51 for HIV, $\geq$5.17 for BVDV, and $\geq$5.75 for BHV. These results indicate that the virus filtration process successfully improved the viral safety of factor IX.

• 한국동물위생학회지
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• 제46권3호
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• pp.193-202
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• 2023

Porcine epidemic diarrhea is an infectious intestinal disease caused by the porcine epidemic diarrhea virus (PEDV). Especially, when suckling piglets are infected, the mortality rate is close to 100%. PEDV is classified into G1 and G2 types based on genetic differences. The G2 type PEDV outbreak in the United States in 2013 was highly pathogenic and contagious, and it has spread worldwide and caused continuous economic losses. Most commercial vaccines used are G1 type vaccines, and existing vaccines do not fully protect piglets due to genetic differences. In this study, we evaluated the safety of the newly developed G2 type attenuated HSGP vaccine strain by inoculating it into piglets and testing whether the vaccine virus spreads to the non-vaccinated, negative pigs and whether the vaccine reverts to its virulence during serial passage experiments. Each experiment lasted for 7 days for each passage, and fecal viral titers, clinical symptoms, and weight gain were measured daily. After the experiment, necropsy was performed to measure intestinal virus titer and pathological evaluation. As a result of the first passage, no transmission of the vaccine virus to negative pigs co-housed with vaccinated pigs was observed. In addition, after four consecutive passage experiments, the clinical symptoms and small intestine lesions were gradually alleviated, and no virus was detected in the feces in the fourth passage experiment. Therefore, it was concluded that the vaccine was safe without virulence reversion in accordance with the guidelines of the current licensing authority. However, further studies are needed on the genetic changes and biological characteristics of the mutant virus that occur during successive passages of the attenuated vaccine since the replication and clinical symptoms of the virus increased until the third passage during successive passages of the vaccine virus. Based on this study, it was concluded that virulence reversion and safety evaluation of attenuated vaccines through serial passage in target animals can be useful to evaluate the safety of attenuated viruses.