Session 1 : 국제적 이슈 인수공통전염병 현황 및 예방대책 ; S1-1 탄저균의 감염과 예방

안종삼 대한인수공통전염병학회 2012 창립총회 및 학술대회 초록집 Vol.2012 No.1

Salmonella A 및 D Group의 특이적 검색을 위한 rfbS 염기서열 분석에 기초한 PCR기법의 적용

안종삼(Jong-sam Ahn),정석찬(Suk-chan Jung),김종만(Jong-man Kim),박용호(Yong-ho Park),우승룡(Seng-ryong Woo),조윤상(Yun-sang Cho),이상운(Sang-un Lee),우희종(Hee-jong Woo) 大韓微生物學會 1996 大韓微生物學會誌 Vol.31 No.2

면역학적인 방법에 의한 한우와 유우의 요네병 발생조사

김종만,안종삼,우승룡,조동희,조윤상,박정문,윤용덕,장국현,Kim, Jong-man,Ahn, Jong-sam,Woo, Seung-roung,Jo, Dong-hee,Jo, Yun-sang,Park, Jeung-moon,Yoon, Yong-dhuk,Chang, Guk-hyun 대한수의학회 1994 大韓獸醫學會誌 Vol.34 No.1

A immunological survey of paratuberculosis in dairy and Korean native cattles was conducted by enzyme linked immunosorbent assay(ELISA), complement fixation test(CFT), agar gel immunodiffusion test (AGID) and intradermal skin test(ID). Over all prevalence of pararuberculosis in cattles was 6.7%(109/1633) by ID, 7.5(205/2719) by AGID, 9.3% (245/2641) by CFT and 13.4%(363/2719) by ELISA. Prevalence in dairy cattle was higher than that of Korean native cattle. Of 70 ELISA-positive cattle, 23(28.6%) and 48(68.6%) cattles were classified as positive in the AGID and positive or suspect in CFT, respectively. Of 92 ELISA-suspect cattle, 32(34.9%) and 48(52.2%) cattles were classified as AGID-positive and CFT-positive or suspect, respectively. It was concluded that paratuberculosis is widespread in cattle of Korea.

Analysis of the seroprevalence of bovine paratuberculosis and the application of modified absorbed ELISA to field sample testing in Korea

박용호,박건택,안종삼,William C. Davis,구혜정,권남훈,정우경,김준만,홍순근 대한수의학회 2006 Journal of Veterinary Science Vol.7 No.4

Paratuberculosis (PTB) is a major disease problem worldwide, and causes major economic losses in the dairy industry. Although PTB has been reported in Korea, no studies have been conducted to determine its prevalence and no program has been developed to control the disease. In this study, the sera of beef (n = 1,056) and dairy cattle (n = 1,105) from all provinces in Korea were tested to determine the prevalence of PTB using two different ELISA: an ‘in house’ modified absorbed ELISA (P-ELISA) based on sonicated antigen from Mycobacterium avium subsp. paratuberculosis ATCC 19698, and a commercial ELISA (C-ELISA). Receiver operating characteristic analysis was used to determine the cutoff point for P-ELISA. Based on C-ELISA results, the area under the curve for P-ELISA was 0.913 (95% CI, 0.883 to 0.943). Using a cutoff point of 0.100, P-ELISA showed a sensitivity of 62.0% and a specificity of 93.7%. The kappa value and the percent agreement between the two ELISAs were 0.322 and 92.5%, respectively. Both ELISAs showed a significant correlation between age and seropositivity (p < 0.01). According to C-ELISA, 71 of 2,161 sera (3.3%, 95 CI, 2.6% to 4.1%) were test-positive. The national true prevalence of PTB was estimated to be 7.1%. The findings suggest that a control program should be implemented to limit the spread of this disease, and that P-ELISA could be used as a screening test that produces results similar to C-ELISA.

장구균의 vancomycin 내성 유전자와 종 특이유전자의 검출을 위한 Multiplex polymerase chain reaction 개발

조윤상,이희수,김종만,안종삼,류판동,박용호,유한상,이문한,Cho, Yun-Sang,Lee, Hee-Soo,Kim, Jong-Man,Ahn, Jong-Sam,Ryu, Pan-Dong,Park, Yong-Ho,Yoo, Han-Sang,Lee, Mun-Han 대한수의학회 2003 大韓獸醫學會誌 Vol.43 No.1

A multiplex PCR assay, which allows simultaneous detection of vancomycin resistant genotypes and Enterococcus species-specific genes, was developed. Vancomycin resistant enterococci (VRE) from chickens and humans could be detected for vanA, vanB, vanC-1, vanC-2, $ddl_{E.faecium}$ and $ddl_{E.faecalis}$ by multiplex PCR. Eight isolates of VRE from humans (n=11) had $ddl_{E.faecium}$ and vanA, and 3 isolates of the VRE had $ddl_{E.faecium}$ and vanB. One isolate of VRE from chickens (n=6) had $ddl_{E.faecium}$ and vanA, and 5 isolates of the VRE had only vanA. E. faecium, E. faecalis, E. gallinarum and E. casseliflavus were also confirmed for the species-specific gene by multiplex PCR. This multiplex PCR could detect E. faecium, E. faecalis, E. gallinarum, E. casseliflavus, vanA, vanB, vanC-1 and vanC-2, simultaneously. The PCR assay established in the present study can be an alternative to time-consuming biochemical tests and antibiotic susceptibility tests of Enterococcus spp.

Lactobacillus reuteri의 Bacillus anthracis Sterne 34 F2 대한 항균효과

권남훈,김소현,배원기,김지연,임지연,노경민,김준만,안종삼,허진,박용호 한국수의공중보건학회 2001 예방수의학회지 Vol.25 No.4

Antimicrobial activities of three different Lactobacillus strains (Lactobacillus reuteri, L. bulgaricus, and L. casei) against Bacillus anthracis were determined on the Mueller Hinton Agar containing each culture supernatant obtained from 3 different growth conditions (MRS without glycerol, MRS with 0.5 M glycerol and 0.25 M glycerol solution).Though antimicrobial activity of L. reuteri in the first two conditions was not better than the others', the activity was significantly higher than those of others in 0.25 M glycerol solution. This prominent effect might be attributable to reuterin, produced by L. reuteri using glycerol. We could detect the presence of reuterin in the supernatant of 0.25 M glycerol solution with 500 MHz Nuclear Magnetic Resonance (NMR). The result of minimum bactericidal concentration (dilution fold) against spore of B. anthracis has revealed that reuterin had strong effect against anthrax spores. To examine any changes of antimicrobial activities of the probiotics, the probiotics were treated with different pH concentration, pepsin or trypsin digestion. The study has revealed that antimicrobial activities of L. bulgaricus and L. casei were significantly decreased, but reuterin activity was not entirely affected by any of these treatments.

비교 피내 검사와 감마 인터페론 검사에 의한 우결핵의 진단

조윤상,김종만,정석찬,우승룡,김종염,유한상,박용호,안종삼,Cho, Yun-sang,Kim, Jong-man,Jung, Suk-chan,Woo, Seung-ryong,Kim, Jong-yeom,Yoo, Han-sang,Park, Yong-ho,Ahn, Jong-sam 대한수의학회 1999 大韓獸醫學會誌 Vol.39 No.6

Since Robert Koch found tubercle bacilli in 1882, the studies on tubercle bacilli of human and animal had been carried out. Being old tuberculin(OT) introduced in 1890, the specificity of the diagnosis of tuberculosis has been improved by continual uses of heat concentrated synthetic medium(HCSM) and purified protein derivatives(PPD) tuberculin. Now, two types of tuberculin test are used worldwidly ; the single intradermal test(SIT) using bovine tuberculin and the single intradermal comparative tuberculin test(SICTT) using avian and bovine tuberculins. In the SICTT, each countries have used with different combination of both avian and bovine tuberculins' titers. However, this kinds of studies have not reported in Korea. Therefore, the studies on the combination of their tuberculins' titers were performed through intradermal test of guinea pigs sensitized with either Mycobacterium bovis or M avium and were examined in 10 cattles of SIT positive reactors. Also, IFN-${\gamma}$ assay, the latest diagnostic method of bovine tuberculosis, was experimentally applied to SIT positive reactors. For determining the optimal titers, sensitized guinea pigs with M bovis and M avium were intradermally injected avian and bovine tuberculin. In guinea pigs sensitized with M bovis, bovine tuberculin 50 T.U. showed significant difference from all tested concentrations of avian tuberculin(p < 0.05). In guinea pigs sensitized with M avium, there is significantly different between bovine tuberculin and avian tuberculin by 25 T.U.(p < 0.01). Therefore, optimal titers of bovine and avian PPD tuberculins' titers for the SICTT in Korea were 5,000 and 2,500 tuberculin units, respectively, and the swelling diffences between bovine and avian site in SIT positive reactors were above 3mm. Also, in IFN-${\gamma}$ assay, the 9 SIT positive reactors were showed all the positive reactions.

Mycobacterium bovis Infection in a Farmed Elk in Korea

김재훈,손현주,강경일,김원일,진영화,안종삼 대한수의학회 2002 Journal of Veterinary Science Vol.3 No.3

Acaseoftuberculosisisreportedinaneight-year-old,male, elk ( Cervus elaphus nelsoni ). The lk showedsevere coughing, respiratory distress, abdominal brea-thing, anorexia, and severe progresive emaciation inthe lk farm. At necropsy, the lk apeared in poor bodycondition. Mild enlargement of retropharyngeal andsubmandibular lymph node was observed in the head.Difuse fibrinous pleuritis and purple red lobar pneu-dark yelow discrete or confluent nodules from 0.3 to2 c m i n d i a m e t e r w e r e s c a t e r e d i n t h e w h o l e l u n g .Bronchial and mediastinal lymph nodes were alsoenlarged. Histopathologically, lungs had typical clasicaltuberculous granulomas, multiple absceses, and numerousmacrophages and Langhans giant cels infiltration inalveolar lumen. In the lymph nodes, there were smalclusters of necrosis and infiltration of numerousmacrophages, epithelioid cels, and Langhans giantcels. With the acid-fast staining, numerous mycobacteriato this study, there are diferences of the histopathologiclesions and the numbers of acid-fast bacilli in thelesions between this elk and catle. Mycobacterium boviswas confirmed as a causative agent in this elk usingbacterial isolation, biochemical characteristics, and PCRtechnique. The isolate was negative for niacin test,nitrate reductase, and pyrazinamidase. This is a firstreport for bovine tuberculosis of armed elk in Asia.

Lactobacillus reuteri의 주요 식품 위해 미생물에 대한 항균 효과

권남훈,김소현,배원기,김지연,임지연,노경민,김준만,안종삼,허진,박용호,Kwon, Nam-Hoon,Kim, So-Hyun,Bae, W.K.,Kim, J.Y.,Lim, J.Y.,Noh, K.M.,Kim, J.M.,Ahn, J.S.,Hur, J.,Park, Y.H. 한국식품위생안전성학회 2001 한국식품위생안전성학회지 Vol.16 No.4

본 연구에서는 5종의 유산균들 (Lactobacillus reuteri, L. acidophillus, L. bulgaricus, L. casei and Bifidobacterium longum)의 식중독 균들에 대한 항균력을 조사하였다. 각 유산균들을 3가지 서로 다른 조건 (MRS+glucose, MRS+0.5 M glycerol, 0.25 M glycerol solution)에서 배양한 후 그 상층액이 포함된 Moulter Hinton Agar에 8종의 유해균들을 접종하였다. MRS+glucose, MRS+0.5 M glycero에서의 상층액을 이용한 실험에서는 L. reuteri의 항균력이 다른 유산균들에 비해 높지 않았으나 0.25 M glycerol solution에서는 탁월하게 높은 결과를 나타내었다. (p<0.05). 0.25 M glycerol solution에서 나타난 높은 항균력은 L. reuteri가 glycerol을 이용하여 생산하는 reuterin이란 물질 때문이라 생각되며, 0.25 M glycerol solution의 상층액을 500 MHz Nuclear Magnetic Resonance (NMR)를 통해 분석한 결과 reuterin의 존재를 확인할 수 있었다. 3가지 조건 하에서의 항균력 시험 결과 가장 높은 항균력을 나타낸 3가지 유산균들 (Lactobacillus resuteri, L. bulgaricus, L. casei)을 선발, 각각 최고의 항균력을 나타낸 조건으로 배양하여 최소사멸농도 (minimum bactericidal concentration)를 측정한 결과, L. reuteri가 생산한 reuterin이 광범위 항균물질임을 확인하였다. 또한 pH 적정, pepsin 혹은 trypsin 처리를 한 후 3가지 유산균들의 항균력 변화를 조사한 결과 L. bulgaricus와 L. casei의 항균력은 이러한 조건에서 상당히 감소한 반면 reuterin의 항균력은 아무런 영향을 받지않았다 (p<0.05). 이상의 결과들로 보아 주요 유산균들 중에서 L. reuteri의 항균력이 가장 우수하면서 광범위하였으며 이는 L. reuteri가 분비하는 항균물질인 reuterin에 기인한 것으로 사료된다. 또한 reuterin의 항균력은 타 유산균과는 달리 장관내 pH나 단백질 분해효소에 의해 영향을 받지 않으므로 인체나 동물 장관에서 가장 높은 효력을 가진다고 판단할 수 있다. Antimicrobial activities of ave different probiotics (Lactobacillus reuteri, L. acidophillus, L. bulgaricus, L. casei and Bifidobacterium longum) against 8 bacterial pathogens were determined on the Mueller Hinton Agar containing supernatant of probiotics obtained from 3 different growth conditions (MRS without glycerol, MRS with 0.5 M glycerol or 0.25 M glycerol solution). Though antimicrobial activity of L. reuteri in the first two conditions was not better than the others', the activity was significantly higher than that of others in 0.25 M glycerol solution. This prominent effect might be attributable to reuterin, produced by L. reuteri using glycerol. We could detect the presence of reuterin in the supernatant of 0.25 M glycerol solution with 500 MHz Nuclear Magnetic Resonance (NMR). The result of minimum bactericidal concentration (dilution fold) has revealed that reuterin showed pan-bactericidal effects against 8 major food-borne pathogens. To examine any changes of antimicrobial activities of the probiotics, the probiotics were treated with different pH conditions, pepsin or trypsin digestion. Antimicrobial activity of reuterin was not entirely affected by any of these treatments, while the activities of the other probiotics were significantly decreased.