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오리에서 발생한 바이러스성 간염과 살모넬라균증의 혼합감염
고바라다,김용환,김계엽 한국수의병리학회 2001 한국수의병리학회지 Vol.5 No.2
Ducklings collected from three farms, having history of rapid onset and spread of nerve sings including kick spasmodically with legs and opisthotonos, were pathologically, bacteriologically, virologically examined. Grossly, multiple petechial to ecchymotic hemorrhages were detected in the swollen liver. Histopathologically, diffuse coagulative necrosis of hepatocytes was characteristic in acute cases. Chronic cases revealed marked bile duct hyperplasia rather than hepatocyte necrosis. Some of these cases exhibited multiple granulomas consisting of macrophages, heterophil, fibrin and necrotic cell debri. Filtered homogenate of livers sampled from ducklings caused embryo death with marked hemorrhage and swollen of liver after inoculation into chorioallantoic membrane. Three strains of Salmonella spp., S montevideo, S hadar, and S give, which were biochemically and serologically identified, were isolated from ducklings of three farms, respectively. From these results, these ducklings were concurrently infected with duck hepatitis virus and Salmonella spp.
Identification of a norovirus from diarrheic dog in Gwangju, Republic of Korea
Ba-Ra-Da Koh,Su-Yeon Seo,Ga-Hoi Choi,Byeong-Cheol Yoon 한국동물위생학회 2023 韓國家畜衛生學會誌 Vol.46 No.3
Noroviruses are a major cause of gastroenteritis in humans and animals worldwide. In 2021, canine norovirus (CNoV) infection was detected at an animal clinic in Gwangju area, South Korea. A seminested polymerase chain reaction was developed to amplify a 478 bp fragment of the RdRp gene of CNoV. The phylogenetic analysis of this fragment confirmed the strain to be genogroup IV.2 (Dog/ GIV.2/gw/s377/2021/KOR), which exhibited the highest similarity to the feline NoV strain GIV.2/ CU081210E/USA/2010 (accession no. NC_045762) with 95.1% nucleotide (nt) identity and 98.7% amino acid (aa) identity. These research findings indicate that the detected norovirus in dogs is genetically similar to a feline-origin norovirus, suggesting easy cross-species transmission among animals.
돼지고기 제품 내 닭고기 검출을 위한 TaqMan® real-time PCR의 적용
고바라다 ( Ba Ra Da Koh ),김지연 ( Ji Yeon Kim ),나호명 ( Ho Myung Na ),박성도 ( Seong Do Park ),김용환 ( Yong Hwan Kim ) 한국동물위생학회 2013 韓國家畜衛生學會誌 Vol.36 No.3
Many consumers are increasingly concerned about the meat they eat, and accurate labelling is important due to public health, economic and legal concerns. Meat species adulteration is a common problem in the retail markets. In this study, a TaqMan® quantitative real-time polymerase chain reaction (PCR) assay was applied for its ability to quantify chicken meat, which was not indicated on the label, in 79 commercial pork products (ham, sausages, bacon and ground meat) producted by 10 different manufacturers. The amplification efficiency was 82.05% and the square regression coefficient (R2) was 0.995. PCR results showed that 38.6% of ham samples, 50.0% of sausages samples, and 50.0% of ground meat samples were contaminated with chicken residuals, while the bacon samples were not contaminated with chicken residuals. Only twelve pork products of one of the manufacturers were in accordance with indicated in their labels. The PCR assay reported in this work could be particularly useful in inspection programs to verify the food labelling of commercial processed meats and to gain consumers` trust.
Mycobacterium bovis 와 M. tuberculosis 감별을 위한 등온증폭법
고바라다 ( Ba Ra Da Koh ),김재명 ( Jae Myung Kim ),성창민 ( Chang Min Sung ),지태경 ( Tae Kyung Ji ),나호명 ( Ho Myung Na ),박성도 ( Seong Do Park ),김용환 ( Yong Hwan Kim ),김은선 ( Eun Sun Kim ) 한국가축위생학회 2013 韓國家畜衛生學會誌 Vol.36 No.2
Mycobacterium (M.) bovis, a member of the M. tuberculosis complex (MTC), is a re-emerging, zoonotic agent of bovine tuberculosis whose prevalence probably depends on variations in direct exposure to cattle and ingestion of raw milk. Accurate species differentiation of M. bovis and M. tuberculosis is needed to distinguish between human and zoonotic tuberculosis. This study successfully developed a loop-mediated isothermal amplification (LAMP) assay for rapid detection and differentiation of M. bovis and M. tuberculosis, however showed negative reactions in eight non-tuberculous mycobacteria (NTM) samples and ten other bacterial species. Sensitivity of this assay for detection of genomic M. bovis DNA was 10 fg/μl. And this assay successfully detected M. bovis in bovine clinical specimens. In conclusion, the LAMP assay is a simple and powerful tool for rapid detection of M. bovis in both pure bacterial culture and in clinical samples.
고바라다 ( Ba Ra Da Koh ),김지연 ( Ji Yeon Kim ),장미선 ( Mi Sun Jang ),서두리 ( Doo Ri Seo ),정보람 ( Bo Ram Jung ),신지현 ( Ji Hyun Shin ),임진택 ( Jin Taek Lim ),김용환 ( Yong Hwan Kim ),김은선 ( Eun Sun Kim ) 한국동물위생학회 2015 韓國家畜衛生學會誌 Vol.38 No.2
A simultaneous determination method was developed for nine preservatives (benzoic acid, sorbic acid, dehydroacetic acid, methyl-, ethyl-, isopropyl-, propyl-, isobutyl- and butyl-parabens) in sausage by liquid chromatography with electrospray ionization tandem mass spectrometry (LC/ESI-MS/MS). Each parameter was established by multiple reaction monitoring in negative mode. Separation was achieved on a phenyl-hexyl (2.5 꺷m, 2.1⊥150 mm, Waters) with A-20 mM ammonium acetate containing 0.1% acetic acid in water, B-Acetonitrile as mobile phase with gradient mode at a flow rate of 0.3 mL/min. The developed method was validated for specificity, linearity, accuracy and precision in sausages samples. Linearity was over 0.998 with calibration curve of the mixed standards. The mean recoveries from sausages fortified at the level of 2.0.10.0 mg/L were in range of 98.60.109.16% with RSDs lower than 8.93%. The limits of detection (LOD) and the limits of quantification (LOQ) were in the range between 0.0003.0.085 mg/L and 0.01.0.257 mg/L, respectively. Intra-day precision and inter-day precision were 0.45.6.16% and 2.81.13.33%, respectively. Using presently developed determination method, 33 field sausage samples from Gwangju city in Korea were screened over nine preservatives. As a result, no preservatives were detected in all samples.
식육감별을 위한 미토콘드리아 12S rRNA와 16S rRNA 유전자의 종 특이적 multiplex PCR 기법 개발
고바라다 ( Ba Ra Da Koh ),김지연 ( Ji Yeon Kim ),나호명 ( Ho Myung Na ),박성도 ( Seong Do Park ),김용환 ( Yong Hwan Kim ) 한국가축위생학회 2011 韓國家畜衛生學會誌 Vol.34 No.4
Species-specific PCR assay was developed for detection of cattle, sheep, goat, horse, dog, pig, chicken, duck, goose, and turkey using mitochondrial 12S rRNA and 16S rRNA as target genes. Also, an internal positive control was used to detect possible false negatives by using 18S rRNA gene. We designed species-specific primers with amplicon length of 190, 219, 350, 467, 241, 119, 171, 229, 111 and 268 bp for cattle, sheep, goat, horse, dog, pig, chicken, duck, goose, and turkey respectively. The specificity of the primers was tested against the other 10 non-target animal species and a cross-reaction was not observed. We developed two multiplex PCR assays for the simultaneous identification of Korea`s major livestock species (cattle, pig, chicken and duck) and poultry species (chicken, duck, goose and turkey) from analogous samples, retaining the same specificity. The limit of detection of the multiplex PCR assay (cattle, pig, chicken and duck) ranged between 1 pg and 0.1 pg of template DNA extracts from raw meat. Applying multiplex PCR assays to DNA extracts from experimental pork/beef and pork/chicken tested raw and heat-treated (120oC for 30 min) mixtures respectively, detection limit was 0.1% level beef in pork, pork in beef and chicken in pork and 1.0% level pork in chicken. In conclusion, this assay using gel-based capillary electrophoresis would be very useful in highly sensitive and rapid identification of animal species or ingredients in minced meat and other meat products.
소 림프절에서 Mycobacterium bovis DNA의 신속 검출과 M. bovis와 M. tuberculosis 감별을 위한 real-time PCR 개발
고바라다 ( Ba Ra Da Koh ),장영부 ( Young Boo Jang ),구복경 ( Bok Kyung Ku ),조호성 ( Ho Seong Cho ),배성열 ( Seong Yeol Bae ),나호명 ( Ho Myung Na ),박성도 ( Seong Do Park ),김용환 ( Yong Hwan Kim ),문용운 ( Yong Un Mun ) 한국가축위생학회 2011 韓國家畜衛生學會誌 Vol.34 No.4
Mycobacterium bovis, a member of the M. tuberculosis complex (MTC), is the causative agent of bovine tuberculosis. Detection of M. bovis and M. tuberculosis using conventional culture- and biochemical- based assays is time-consuming. Therefore, a simple and sensitive molecular assay for rapid detection would be of great help in specific situations such as faster diagnosis of bovine tuberculosis (bTB) infection in the abattoirs. We developed a novel multiplex real-time PCR assay which was applied directly to biological samples with evidence of bTB and it was allowed to differentiate between M. bovis and M. tuberculosis. The primers and TaqMan probes were designed to target the IS1081 gene, the multi-copy insertion element in the MTC and the 12.7-kb fragment which presents in M. tuberculosis, not in the M. bovis genome. The assay was optimized and validated by testing 10 species of mycobacteria including M. bovis and M. tuberculosis, and 10 other bacterial species such as Escherichia coli, and cattle lymph nodes (n=113). The tests identified 96.4% (27/28) as M. bovis from the MTC-positive bTB samples using conventional PCR for specific insertion elements IS1081. And MTC-negative bTB samples (n=85) were tested using conventional PCR and the real-time PCR. When comparative analyses were conducted on all bovine samples, using conventional PCR as the gold standard, the relative accuracy of real-time PCR was 99.1%, the relative specificity was 100%, and the agreement quotient (kappa) was 0.976. The detection limits of the real-time PCR assays for M. bovis and M. tuberculosis genomic DNA were 10 fg and 0.1 pg per PCR reaction, respectively. Consequently, this multiplex real-time PCR assay is a useful diagnotic tool for the identification of MTC and differentiation of M. bovis and M. tuberculosis, as well as the epidemiologic surveillance of animals slaughtered in abattoir.
광주지역에서 즉석 제조ㆍ판매하는 식육가공품의 안전성 조사
고바라다 ( Ba-ra-da Koh ),서은주 ( Eun-ju Seo ),안아진 ( Ah-jin Ahn ),정보람 ( Bo-ram Jung ),하이든 ( Yi-deun Ha ),서두리 ( Doo-ri Seo ),임진택 ( Jin-taek Lim ),김용환 ( Yong-hwan Kim ),김은선 ( Eun-sun Kim ) 한국가축위생학회 2017 韓國家畜衛生學會誌 Vol.40 No.1
The main goal of this survey was to assess the current sanitation status and safety standards of meat processed products purchased at instant meat sales and processing operators. Analyses were carried out from April to September in 2016 in Gwangju area, Republic of Korea. A total number of 150 samples including seasoned meats, ground meat products, meat extract products, heated seasoned meats, sausages and hams from 35 butcher shops was collected. The number of inappropriate cases was revealed eleven cases (7.3%) in total viable count of bacteria (TVC), total coliform counts (TCC) and Listeria monocytogenes. The reported data indicate that more systemic and technical guidance is needed to monitor instant meat sales and processing operators in order to guarantee safety of meat processed products.
고바라다 ( Ba Ra Da Koh ),김현중 ( Hyun Joong Kim ),박덕웅 ( Duk Woong Park ),박성도 ( Seong Do Park ),김재익 ( Jae Ik Kim ),박종태 ( Jong Tae Park ),김용환 ( Yong Hwan Kim ) 한국가축위생학회 2007 韓國家畜衛生學會誌 Vol.30 No.3
Bovine tuberculosis is an important zoonosis worldwide. Mycobacterium bovis, the causative agent of this disease in cattle, is also a pathogen for humans and several economically important animals. The cases of tuberculosis are reported in two cow found at slaughter house located in Gwangju city. Histopathologically, in the lymph nodes, granulomas consisted of large areas of necrosis surrounded by variable thick bands of cellular infiltrate containing macrophages, Langhans-type multinucleated giant cells and lymphocytes. Lesions in the lung followed the same developmental pattern as did lesions in the lymph nodes with some exceptions. With the acid-fast staining, numerous mycobacteria were revealed in the lung and lymph nodes. M bovis was confirmed as a causative agent in these cattle using bacterial isolation and PCR and restriction fragment length polymorphism method based on a unique 12.7kb fragment insertion sequence from the Mycobacterium tuberculosis genome and the pncA polymorphism. The insertion element IS6110 and IS1081 were present M bovis isolated from lungs and lymph nodes of cattle using PCR assay. These cases are interesting and important in public health aspect that M bovis-infected cattle were found during a routine post-mortem inspection at slaughter house.
광주지역 동물보호소내 유기견의 개심장사상충과 개 브루셀라병 감염 실태조사
고바라다 ( Ba Ra Da Koh ),나호명 ( Ho Myung Na ),장미선 ( Mi Sun Jang ),김지연 ( Ji Yeon Kim ),박성도 ( Seong Do Park ) 한국가축위생학회 2007 韓國家畜衛生學會誌 Vol.30 No.1
This study was conducted to investigate the prevalence of canine heartworm infections, canine brucellosis and hematologic values from 153 free roaming dogs in the area of Gwangju city from March to November 2006. Nineteen (12.4%) of 153 samples tested with modified Knott`s technique showed positive reaction for microfilariae. Polymerase chain reaction using specific primers for D immitis amplified the expected product from all samples of 19 microfilaremic canine blood samples as determined by the modified Knott`s test for microfilariae. The seasonal infection rates of microfilariae were higher in the spring season (10/19, 52.6%) than in the other seasons. The major hematological findings in microfilaremic dogs were mild leukocytosis and mild monocytosis. A total of 100 dogs randomly selected from 153 free roaming dogs were negative for canine brucellosis by serological test using immunochromatographic antibody test kit.