• Title/Summary/Keyword: Animal and Plant Quarantine Agency

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Expression of the VP2 protein of feline panleukopenia virus in insect cells and use thereof in a hemagglutination inhibition assay

  • Yang, Dong-Kun;Park, Yeseul;Park, Yu-Ri;Yoo, Jae Young;An, Sungjun;Park, Jungwon;Hyun, Bang-Hun
    • Korean Journal of Veterinary Research
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    • v.61 no.2
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    • pp.19.1-19.7
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    • 2021
  • Feline panleukopenia virus (FPV) causes leukopenia and severe hemorrhagic diarrhea, killing 50% of naturally infected cats. Although intact FPV can serve as an antigen in the hemagglutination inhibition (HI) test, an accidental laboratory-mediated infection is concern. A non-infectious diagnostic reagent is required for the HI test. Here, we expressed the viral protein 2 (VP2) gene of the FPV strain currently prevalent in South Korea in a baculovirus expression system; VP2 protein was identified by an indirect immunofluorescence assay, electron microscopy (EM), Western blotting (WB), and a hemagglutination assay (HA). EM showed that the recombinant VP2 protein self-assembled to form virus-like particles. WB revealed that the recombinant VP2 was 65 kDa in size. The HA activity of the recombinant VP2 protein was very high at 1:215. A total of 143 cat serum samples were tested using FPV (HI-FPV test) and the recombinant VP2 protein (HI-VP2 test) as HI antigens. The sensitivity, specificity, and accuracy of the HI-VP2 test were 99.3%, 88.9%, and 99.3%, respectively, compared to the HI-FPV test. The HI-VP2 and HI-FPV results correlated significantly (r = 0.978). Thus, recombinant VP2 can substitute for intact FPV as the serological diagnostic reagent of the HI test for FPV.

Evaluation of Japanese encephalitis virus vaccine strains currently used in pigs by molecular characterization

  • Lee, Jeong-Ah;Yang, Dong-Kun;Kim, Ha-Hyun;Kim, Sun-Young;Nah, Jin-Ju;Cho, Soo-Dong;Song, Jae-Young
    • Korean Journal of Veterinary Service
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    • v.35 no.3
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    • pp.169-174
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    • 2012
  • Japanese encephalitis virus (JEV) is one of the main causes of viral encephalitis in human and animals. For over 30 years, a live attenuated JEV vaccine strain has been used in the veterinary field, and it is required to conduct quality evaluation studies on the commercial vaccines. For the quality control of live attenuated JEV vaccine, we investigated the nucleotide sequence similarity of prME gene derived from five JEV vaccines commercially available in pigs in Korea. The Vero cells infected with JEV vaccines showed specific cytopathic effect, which was characterized by rounding and detached cells. In the phylogenetic analysis, all of the vaccine strains showed a close relationship with the original vaccine seed strain (Anyang 300) and clustered into the genotype 3. In comparison of the nucleotide and deduced amino acid sequences of prME genes with the original strain, all JEV vaccine strains showed high amino acid similarity ranging from 98.9% to 99.5%, but had several point mutations, probably due to high mutation rates of viral RNA polymerase by several virus passages. Even though the current JEV vaccine strains have been maintained and produced for a long period of time, the genetic characterization of them have been rarely changed. However, since the mid 1990's, molecular epidemiology of JEV has been changed sharply from genotype 3 to genotype 1 in Korea, further studies on new vaccine strains to genotype 1 is required for more effective prevention in the field.

Infection of Foot and Mouth Disease of Serotype A in Farm-raised Deer in Korea (국내 사슴에서 구제역 A형 감염증례)

  • Park, Jong-Hyeon;Lee, Kwang-Nyeong;Kim, Su-Mi;Ko, Young-Joon;Lee, Hyang-Sim;Cho, In-Soo;Kim, Byoung-Han
    • Journal of Veterinary Clinics
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    • v.29 no.4
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    • pp.315-318
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    • 2012
  • This report describes a case of foot and mouth disease (FMD) in farm-raised Sika deer. FMD serotype A occurred in six cattle farms in January 2010 since the outbreak of pig cases 2002 in South Korea. In addition to the six cattle farms where the disease occurred during January, positive reactions to FMD antibodies were found in two heads of deer on March at a deer farm in serological tests intended to lift the movement restriction of the susceptible animals imposed within risk zone. In the specimens collected from 12 heads for confirmatory tests in the same farm, no virus was detected in antigen tests and neutralising antibody titers in all raising deer in the farm were relatively high. So it seems likely that the animals had been infected with FMD at least one month earlier when they were found.

Development of high-performance liquid chromatography methods for the anticoccidials: toltrazuril and diclazuril (고속액체크로마토그래피를 이용한 항원충제 분석법 개발: 톨트라주릴 및 디클라주릴)

  • Jeong, Kyung Hun;Jeong, Miyoung;Park, Hae-Chul;Hossain, Md Akil;Kim, Daegyun;Lee, Kwang-Jick;Kang, Jeong Woo
    • Korean Journal of Veterinary Research
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    • v.57 no.4
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    • pp.223-226
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    • 2017
  • This study was undertaken to develop new analytical methods for assessment of anticoccidials. High-performance liquid chromatography (HPLC) was found to be a fast, reliable, and practical method. The anticoccidials used in this experiment were toltrazuril and diclazuril, and the analysis factors were specificity, linearity, accuracy, repeatability, and intermediate precision. The linearity of each anticoccidial was better than 0.99, and the accuracies were 99.5% and 99.1% with relative SD of 0.5 and 0.4, respectively. To assess whether the developed HPLC method could be effectively applied, toltrazuril and diclazuril post-market veterinary products (five products) that are currently sold were tested. The results revealed no non-compliant items and the method was applied successfully. Therefore, the newly developed HPLC method for anticoccidial assessment described in this study may be useful as a reference method in the Korean Standards of Veterinary Pharmaceuticals for the analysis of toltrazuril and diclazuril.

Single-nucleotide polymorphism-based epidemiological analysis of Korean Mycobacterium bovis isolates

  • Kim, Tae-Woon;Jang, Yun-Ho;Jeong, Min Kyu;Seo, Yoonjeong;Park, Chan Ho;Kang, Sinseok;Lee, Young Ju;Choi, Jeong-Soo;Yoon, Soon-Seek;Kim, Jae Myung
    • Journal of Veterinary Science
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    • v.22 no.2
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    • pp.24.1-24.16
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    • 2021
  • Background: Bovine tuberculosis (TB) is caused by Mycobacterium bovis, a well-known cause of zoonotic tuberculosis in cattle and deer, and has been investigated in many physiological and molecular studies. However, detailed genome-level studies of M. bovis have not been performed in Korea. Objectives: To survey whole genome-wide single-nucleotide polymorphism (SNP) variants in Korean M. bovis field isolates and to define M. bovis groups in Korea by comparing SNP typing with spoligotyping and variable number tandem repeat typing. Methods: A total of 46 M. bovis field isolates, isolated from laryngopharyngeal lymph nodes and lungs of Korean cattle, wild boar, and Korean water deer, were used to identify SNPs by performing whole-genome sequencing. SNP sites were confirmed via polymerase chain reaction using 87 primer pairs. Results: We identified 34 SNP sites with different frequencies across M. bovis isolates, and performed SNP typing and epidemiological analysis, which divided the 46 field isolates into 16 subtypes. Conclusions: Through SNP analysis, detailed differences in samples with identical spoligotypes could be detected. SNP analysis is, therefore, a useful epidemiological tracing tool that could enable better management of bovine TB, thus preventing further outbreaks and reducing the impact of this disease.

Re-evaluation on conversion system of BactoScan and BactoCount for raw milk in South Korea (국내 원유 세균수 검사장비 박토스캔 및 박토카운트의 보정식 재평가)

  • Kim, ESeul;Kim, Jin-Hwan;Byun, Yeong-Seob;Park, Dasom;Kim, Ha-Young;Yoon, Soon-Seek;Moon, Jin-San
    • Korean Journal of Veterinary Service
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    • v.45 no.1
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    • pp.39-45
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    • 2022
  • The total bacteria count is significantly important factor for hygienic quality in raw milk. BactoScan FMTM and BactoCount IBCTM are the automated instruments for the determination of the total bacterial count in raw milk. They have been used after calibration by standard plate count (SPC) method in South Korea since 2000. It is necessary to re-evaluate for total bacterial counter according to the improvement of milk quality and the change of milk quality grade. Therefore, this study was evaluated the conversion mode of the two machines by SPC method. We collected 921 bulk-tank milk samples throughout the concentration range of 1,000~1,000,000 CFU/mL from June 2020 to October 2021. As a result, when compared by the SPC value, there was a slight difference in total bacterial count in BactoScan below 10,000 CFU/mL and above 200,000 CFU/mL and in BactoCount above 100,000 CFU/mL, respectively. Therefore, the conversion factor for BactoScan and the conversion equation for BactoCount were newly adjusted based on SPC value, and then the correlation coefficients (R2) was 0.85 or higher. In addition, the correlation (R2) between BactoScan and BactoCount was 0.91, which means the results were high positive correlation. These results are expected to contribute to improving the accuracy of the automated instruments for determining of total bacterial count in raw milk.

Differentiation potential of canine mesenchymal stem cells on hydrogel scaffold-based three-dimensional environment (하이드로젤 지지체 기반 3차원 환경에서 개 간엽줄기세포의 분화능 분석)

  • Gu, Na-Yeon;Park, Mi Jeong;Lee, Jienny;Byeon, Jeong Su;Jeong, Da-Un;Cho, In-Soo;Cha, Sang-Ho
    • Korean Journal of Veterinary Research
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    • v.58 no.4
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    • pp.211-217
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    • 2018
  • Mesenchymal stem cells (MSCs) are useful candidates for tissue engineering and cell therapy. Physiological cell environment not only connects cells to each other, but also connects cells to the extracellular matrix that provide mechanical support, thus exposing the entire cell surface and activating signaling pathways. Hydrogel is a polymeric material that swells in water and maintains a distinct 3-dimensional (3D) network structure by cross linking. In this study, we investigated the optimized cellular function for canine adipose tissue-derived MSCs (cAD-MSCs) using hydrogel. We observed that the expression levels of Ki67 and proliferating cell nuclear antigen, which are involved in cell proliferation and stemness, were increased in transwell-hydrogel (3D-TN) compared to the transwell-normal (TN). Also, transforming growth factor-${\beta}1$ and SOX9, which are typical bone morphogenesis-inducing factors, were increased in 3D-TN compared to the TN. Collagen type II alpha 1, which is a chondrocyte-specific marker, was increased in 3D-TN compared to the TN. Osteocalcin, which is a osteocyte-specific marker, was increased in 3D-TN compared to the TN. Collectively, preconditioning cAD-MSCs via 3D culture systems can enhance inherent secretory properties that may improve the potency and efficacy of MSCs-based therapies for bone regeneration process.

Development of Diagnostic Technology of Xylella fastidiosa Using Loop-Mediated Isothermal Amplification and PCR Methods

  • Kim, Suyoung;Park, Yujin;Kim, Gidon
    • Research in Plant Disease
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    • v.27 no.1
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    • pp.38-44
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    • 2021
  • Xylella fastidiosa is the most damaging pathogen in many parts of the world. To increase diagnostic capability of X. fastidiosa in the field, the loop-mediated isothermal amplification (LAMP) and polymerase chain reaction (PCR) assay were developed to mqsA gene of citrate-synthase (XF 1535) X. fastidiosa and evaluated for specificity and sensitivity. Both assays were more robust than current published tests for detection of X. fastidiosa when screened against 16 isolates representing the four major subgroups of the bacterium from a range of host species. No cross reaction with DNA from healthy hosts or other species of bacteria has been observed. The LAMP and PCR assays could detect 10-4 pmol and 100 copies of the gene, respectively. Hydroxynaphthol blue was evaluated as an endpoint detection method for LAMP. There was a significant color shift that signaled the existence of the bacterium when at least 100 copies of the target template were present.

Forensic analysis of toxic substances in fatalities with suspected companion animal cruelty (반려동물 학대 의심 폐사축에 대한 중독물질검사 연구)

  • JeongWoo Kang;Ah-Young Kim;Hyun Young Chae;Hanae Lim;Suncheun Kim;Bok-Kyung Ku;Kyunghyun Lee
    • Korean Journal of Veterinary Research
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    • v.63 no.3
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    • pp.21.1-21.6
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    • 2023
  • The increasing prevalence of toxic substance-exposure in pets in South Korea endangers the health and safety of numerous companion animals, and has become a cause for concern. Notably, the annual incidence of forensic analysis in pets has increased by more than 150% in South Korea, mainly in populous regions such as Seoul, Incheon, and Gyeonggi. In response to this growing issue, veterinary forensic examinations were conducted on 549 dogs and cats from 2019 to 2022. This study revealed the presence of various toxic substances, including pesticides, insecticides, and drugs such as analgesics, anesthetics, antidepressants, and muscle relaxants, in pets. Among the 38 different toxins identified in pets, coumatetralyl, methomyl, terbufos, and buprofezin were the most frequently detected. In this study, toxic substances for pets were identified based on the "toxic agent list for humans," developed by the National Forensic Services, because no list of toxic agents for animals currently exists and data regarding potentially toxic substances for dogs and cats is limited. This is one of the limitations of this study, and necessitates the establishment of a toxic agent list for animals. Continued monitoring and research is also recommended to reveal the incidence, causes, and solutions of toxicity in animals.

Radiation safety management for diagnostic radiation generators and employees in animal hospitals in Korea (동물병원의 진단용 방사선 발생장치 및 방사선종사자 안전관리 실태 조사)

  • An, Hyo-Jin;Kim, Chung-Hyun;Kwon, Young-Jin;Kim, Don-Hwan;Wee, Sung-Hwan;Moon, Jin-San
    • Korean Journal of Veterinary Research
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    • v.54 no.3
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    • pp.151-157
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    • 2014
  • A nationwide survey on radiation safety management in Korean animal hospitals was conducted. By 2013, 53 radiation generators were registered as veterinary medical devices (41 X-ray generators and 12 computed tomography scanners). Additionally there were six approved laboratories for radiation equipment and protection facility, and five approved laboratories for radiation exposure of employees, respectively. By March 2013, 2,030 out of 3,829 animal hospitals operated radiation-generating devices. Among these devices, 389 (19.2%) out of 2,030 were not labeled with the model name and 746 (36.7%) were not labeled with production dates. Thus, most veterinary X-ray generators were outdated (42.6%) and needed replacements. When periodic inspections of 2,018 animal hospitals were performed after revision of the Veterinarians Act in 2011, the hospitals were found to be equipped with appropriate radiation generators and protection facilities. Among 2,545 employees exposed to radiation at the hospitals, 93.9% were veterinarians, 4.3% were animal nurse technicians, and 18% held other positions. Among 169 employees supervised by administrators, none of those had a weekly maximum operating load that exceeded $10mA{\cdot}min$. This study suggests that the radiation safety management system of animal hospitals was general good.