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도계처리 단계별 도체와 처리수의 세균오염 및 염소처리 효과
이철현 ( Chul Hyun Yi ),변유성 ( You Sung Byun ),황보원 ( Bo Won Hwang ),조광제 ( Kwang Je Cho ),강호조 ( Ho Jo Kang ) 한국동물위생학회 1997 한국동물위생학회지 (KOJVS) Vol.20 No.2
This study was carried out assess the effect of the chlorine treatment into water for processing chicken products in each stage of slaughtering, with a special viewpoint related reducing the viable number of microorganisms by which the water and the chicken body were contaminated. The mean bacterial number on chicken samples after picking process was log5.37±0.20~5.84±0.16CFU/㎠. When assessed by standard plate count method, it was the higher one than any other processing stage in which eviscerating, pinning, packaging, and chilling was followed in order of the mean bacterial number. The coliform bacterial numbers on carcasses after sampling from different processing stages were log2.11±0.63~2.88±0.25MPN㎠, which show almost similar numbers in each processing stage. But, after chilling process the number was decreased slightly. The bacterial counts in the water for scalding and chilling showed log3.43±0.59~5.06±0.21 and log 4.30±0.21~6.62±0.33CFU/ml, respectively. In the coliform counts for the water taken out from the 2nd chilling tank, the number was log1.97±0.35~2.91±0.22MPN/ml which showed higher than those of the 1st and the 3rd chilling tank water. The effect of chlorination in reducing the bacterial numbers was accepted at the residual chlorine concentration of 1mg/l by showing the reduction from 108 to 104CFU level and the numbers were decreased less than 10CFU at the concentration of 5mg/l, when assessed by viable cell counts. In conclusion, these results suggested that chlorination in chilling water with final concentration of 5mg/l was strongly recommended to reduce the bacterial numbers on final chicken products.
닭고기에서 병원성 및 변질미생물의 감소를 위한 염소와 유산의 병용처리 효과
이철현 ( Chul Hyun Yi ),변유성 ( You Sung Byun ),황보원 ( Bo Won Hwang ),강호조 ( Ho Jo Kang ) 한국동물위생학회 1999 한국동물위생학회지 (KOJVS) Vol.22 No.4
In this studies, the ability of chlorine and lactic acid to reduce bacterial population of the pathogenic microorganisms were examined on artificially inoculated chicken skin. About 10(5) cells of staphylococcus aureus, salmonella enteritidis, listeria monocytogenes and escherichia coli O157:H7 were inoculated in chicken skin. The contaminated samples were washed for 1 min with sodium hypochlorite solutions that contained 2, 5, 10, 20 and 50mg/ℓ available chlorine and counted number of the agents. Viable population were no significantly difference (p ≥ 0.05) between concentration of chlorine and strains of the pathogens. In the samples inoculated with pathogens were washed in 20mg/ℓ chlorine and then stored at 5°C for up to 10 days, the initial counts of psychrotrophs and aerobic plate counts were 4.02 to 4.36 log cfu/cm2 and increased slightly in course of time. But 10 days after, the pathogens were a little reduced from 3.66~4.91 log cfu/cm2 to 2.54~4.66 log cfu/cm2. In the case of washed skin with solution of 20mg/ℓ chorine and 0.5% lactic acid then store at 5°C for up to 10 days, population of psychrotrophs and aerobic plate counts on chicken skin were markedly reduced immediately after treatment, but the numbers of contaminants were slightly increased after 6 and 8 days. Specifically, numbers of St aureus, S enteritidis, L monocytogenes and E coli 0157:H7 were reduced to 0.5, 0.4, 0.3 and 1.15 log cfu/cm2 after 10 days of storage, respectively, on aerobic plate counts.
원유로부터 Listeria monocytogenes의 신속검색을 위한 종합효소 연쇄반응법의 개선
이철현,손원근,강호조,Yi, Chul-hyun,Son, Won-geun,Kang, Ho-jo 대한수의학회 1996 大韓獸醫學會誌 Vol.36 No.1
The present study was conducted to rapidly detect Listeria monocytogenes in raw milk. Specificity and sensitivity of polymerase chain reaction(PCR) technique, and direct PCR were examinded in raw milk, also were compared the calssical culture methods with PCR technique. This method used a pair of primers based on a unique region in the 16S rRNA sequence of L nomocytogenes. In the PCR specificity tests, each of the 10 strains of L monocytogenes tested gave a single 70-bp band. But the other six Listera spp tested gave negative results. Results of the sensitivity tests showed that as few as 2 CFU of L monocytogenes in pure cultures could be detected with 16S rRNA-based primers, L-1 and L-2. In different PCR cycles, a PCR product was detected with $10^3$ cells of L monocytogenes from 25 cycles to 50 cycles and the concentration of PCR products was cycle-dependent. Raw milk samopes added L monocytogenes cells gave negative results. However, these samplers gave a single 70-bp band by pretreatment of pronase, and PCR products were detected with $10^1$ cells of L monocytogenes. To detemine the most sensitive culture protocol to use in conjunction with the PCR assay, raw milk samples were inoculated with L monocytogenes at concentrations ranging from 1 to $5.7{\times}10^4CFU/ml$. PCR assays from Listeria enrichment broth(LEB) containing raw milk samples added L monocytogene EGD could dtect 10 cells in pronase-pretreated samples without incubation, and 1 cell of L monocytogenes in both 12 hr and 24 hr incubation, respectively. Isolation raw of PCR assays was similar to that of classical culture methods, but required time for detection of L monocytogenes could remarkably be reduced compare to culture methods.
우용구,이수화,이철현,이오수,김봉환,Woo, Yong-Ku,Lee, Su-Hwa,Yi, Chul-Hyun,Lee, O-Soo,Kim, Bong-Hwan 대한수의학회 2003 大韓獸醫學會誌 Vol.43 No.1
Pullorum disease due to Salmonella enterica subspecies enterica bioserovar Pullorum (S. pullorum) is reported to be an endemic disease in domestic poultry flocks. The pulsed-field gel electrophoresis (PFGE) subtyping method was used to assess the extent of genetic diversity and clonality of most of salmonella serotypes and other diverse bacterial species from animals and environmental samples in worldwide. Nowadays, PFGE has already been evaluated as a gold standards for molecular subtyping of salmonella serotypes compared with other molecular analysis methods. PFGE of XbaI digested chromosomal DNA from 23 strains of S. pullorum gave 5 distinctive pulsotypes (from SXPI to SXPV) with 5% confidence range of Dice coefficients, indicating that PFGE is very discriminative and that multiple clones of S. pullorum have been existed and diffused all of domestic poultry flocks industries since 1995. Two dominant pulsogroups (SXA & SXB) appeared as a major clones in this country, because they had consistently been recovered from diverse sources including both chicken organs and raw feed materials between 1995 and 1998. In addition, the matching percentage of PFGE profiles (PFP) among strains from both chickens and feed ingredients provides indirect evidence of the possible transmission of pullorum disease from contaminated raw feed ingredients for chicken production. In calculating of discrimination index (DI) for PFGE method by Simpson's index, DI was appeared as 0.917. Therefore, this index suggested that the present PFGE would seem to be a desirable and confident molecular typing method for S. pullorum strains. To our knowledge for pullorum disease, this is the first study to compare S. pullorum strains from chicken organs and feed samples using the PFGE.
개 코로나바이러스 불활화 백신에 대한 개와 기니픽 간의 면역반응 비교
안동준,김병한,정병열,이철현,전우진,이필수,정갑수,An, Dong-jun,Kim, Byoung-han,Jung, Byeong-yeal,Yi, Chul-hyun,Jeon, Woo-jin,Lee, Pil-soo,Chung, Gab-soo 대한수의학회 2005 大韓獸醫學會誌 Vol.45 No.2
Canine coronavirus (CCV) causes a mild gastroenteritis in dogs. The virus is highly contagious. Although the virus was isolated more than thirty years ago, canine coronavirus infection continues to be a widespread problem. Mixed infections with both CCV and canine parvovirus (CPV) are common. Four kinds of commercial killed CCV vaccines are available in Korea. All the commercial vaccines should pass the National Assay for Veterinary Biologicals prior to release. For the potency test of CCV vaccine, it is necessary to use CCV antibody free dogs. The test requires not only kennels but high cost. To develop easy, efficient and economic potency test method for killed CCV vaccine using laboratory animals, a series of experiments with rabbits and guinea pigs were carried out in this study. In the preliminary test, the guinea pigs showed better immune responses than rabbits. The guinea pig was also easy to manage. So guinea pig was selected for the potency test animals. When the guinea pigs were inoculated twice with one dose of vaccine intramuscuarly each, slower and a little lower SN antibody titers were induced in guinea pigs than in dogs (about 2 kg body weight Beagle strain) given the same posology as guinea pigs'. It was concluded that guinea pigs could be substituted for dogs in the potency test of killed CCV vaccine.
동물용 생 바이러스 백신에서 Mycoplasma 검출을 위한 PCR 기법 적용
전우진,김병한,정병열,안동준,이철현,장환,정갑수,Jeon Woo-Jin,Kim Byoung-Han,Jung Byeong-Yeal,An Dong-Jun,Yi Chul-Hyun,Jang Hwan,Chung Gab-Soo 한국미생물학회 2005 미생물학회지 Vol.41 No.4
동물용 생 바이러스 백신 내에 mycoplasma를 검출하기 위해 polymerase chain reaction (PCR)기법과 2가지의 상품화된 PCR 검출킷트를 평가하였다. PCR기법은 시험에 사용된 모든 mycoplasma를 특이적으로 검출할 수 있었으나, 2가지의 상품화된 PCR 검출킷트는 일부의 mycoplasma를 검출하지 못하였다. 또한, PCR기법의 검출 특이도는 조류 유래 mycoplasma에 속한 4주의 표준주 및 7주의 야외분리주를 모두 검출할 수 있었다. PCR기법의 민감도는 9 CFR Mycoplasma액체배지에서 배양한 Mycoplasma 속균 및 Acholeplasma속균에 대해 $1\~100$ colony forming units/ml까지 검출할 수 있었다. 동물용생 바이러스 백신에 대해 PCR기법의 적용가능성을 평가하기 위해, 돼지 전염성위장염 및 로타바이러스 흔합백신과 개 파보바이러스 백신내에 A. laidlawii를 인공적으로 접종한 후, PCR기법의 민감도를 조사하였을 때 배양액을 이용한 검출한계와 유사하였다. 본 연구에서 사용된 PCR 기법은 동물용 생 바이러스 백신내의 mycoplasma를 신속하고 민감하게 검출할 수 있을 것으로 판단되었다. We evaluated the PCR assay and two commercialized PCR kits for the detection of mycoplasma in veterinary via live vaccines. The PCR assay could specifically detect all the tested Mycoplasma spp. and Acholeplasma spp., whereas two commercialized PCR kits did not. Also, the specificity of the PCR assay showed that 4 reference strains and 7 field isolates belonging to avian mycoplasma species could be all detected. The sensitivity of the PCR assay was determined using pure cultured Mycoplasma spp. and Acholeplasma spp. with a range of 1 to 100 colony forming units/ml in 9 CFR Mycoplasma broth. To test the availability of the PCR assay for veterinary live viral vaccines, A. laidlawii was artificially inoculated into the swine transmissible gastroenteritis-rota virus combined vaccine and canine parvovirus vaccine, respectively and the sensitivity of the PCR assay was similar with the result of cultured samples. In this study, the PCR assays could be used as rapid and sensitive methods for the detection of mycoplasma in veterinary live viral vaccines.