중합효소연쇄반응을 이용한 한우에 감염된 Theileria sergenti의 신속한 검출

채준석,이주묵,권오덕,박진호,채건상,Chae, Joon-seok,Lee, Joo-mook,Kwon, Oh-deog,Park, Jin-ho,Chae, Keon-sang 대한수의학회 1996 大韓獸醫學會誌 Vol.36 No.1

To make the genomic DNA probe of Theileria sergenti, the merozoites were purified from erythrocytes of Korean cattle, The previous studies on the probe of T sergenti had resulted in two probes as KTS1 and KTS3 DNA fragment. Nucleotide sequence of both ends of the KTS1 and KST3 were determined in order to design primers for polymerase chain reaction. A pair of an uper primer(5'-CCTCTTGAAGTCATCCATGT-3'; nucleotide position 48) and a lower primer(5'-CACTGAGCTG GAAAGAGCTA-3'; nucleotide position 156) in pKTS1 were synthesized. The anticipated PCR product was 128bp in length. To examine the sensitivity of the PCR, KTS1 DNA and purified T sergenti DNA were serially diluted by tenfolds with distilled water. The primers were sensitive enough to detect 4ag of the authentic template DNA and 4fg of the purified T sergenti DNA by PCR. Furthermore, when the blood was serially diluted by two-folds with 0.9% saline, the pair could detect up to 0.00029%(about 164 parasites in $10{\mu}l$ of blood) of T sergenti infection in bovine erythrocytes by PCR. In a comparison of microscopic and PCR detection of T sergenti in the same samples from Chonbuk area, 47 and 51 out of 70 sample(67.1%) were positive by the former and by the latter method, respectively.

개의 Babesia gibsoni 감염예방에 관한 연구 1. 항원의 Sonication 및 Formalin 처리에 의한 예방접종

채준석,인동철,이주묵,윤창모,Chae Joon-Seok,Ihn Dong-Cheol,Lee Joo-Muk,Yoon Chang-Mo 한국임상수의학회 1990 한국임상수의학회지 Vol.7 No.1

To examine the effects of vaccination against Babesia gibsoni infection in dogs, 15 normal mixed-breed dogs(5 month to 1 year old) divided into 3 groups with 5 dogs in a group. One of them was selected as control group(group A) and others were selected as experimental groups(group B and C). The group B was vaccinated with sonicated antigens and the group C was vaccinated with 0.2% of formalin treated antigens. The results obtained in the examination were summarized as follows : 1. In the western blot, the lane A revealed specific two bands on the regions of 54kd and 100kd, respectively. 2. After the first vaccination, the antibody titers of group B and C were higher 5 times(1 : 200) than those of control group(1:40). After the second vaccination, the antibody titers of group B and C have not changed. When challenged with the protozoa(Babesia gibsoni), the antibody titers(1 : 5,000) were elevated in all groups. But these were not exceeded over 1 : 5,000 for 4 weeks. 3. After challenge, the peak time of increased numbers of the protozoa was the 15th day (12-18 days) in all groups. During these days, the rate of parasitized erythrocytes in control group was 55.0${\pm}$5.4%. But those of group B and group C were 26.0${\pm}$6.4%, and 15.6${\pm}$7.8%, respectively. 4. After challenge, all of the values of PCV, Hb, RBC were shown to decrease in all of the control and experimental groups. 5. The total leukocytes counts are shown a tendency of reduction in all groups after challenge. 6. In all groups, there were increase in lymphocytes and monocytes after challenge.

한우에 감염된 Theileria sergenti merozoite의 순수분리와 genomic DNA probe에 관한 연구

채준석,이주묵,권오덕,채건상,Chae, Joon-seok,Lee, Joo-mook,Kwon, Oh-deog,Chae, Keon-sang 대한수의학회 1994 大韓獸醫學會誌 Vol.34 No.2

To make the genomic DNA probe of Theileria sergenti, the merozoites were purified from bovine erythrocytes. The infected erythrocytes were lysed by Aeromonas hydrophila(Ah-1) hemolysin, and the parasites were isolated by ultracentrifugation on a Percoll discontinuous density gradient. For construction of a T sergenti genomic DNA library, T sergenti DNA was digested with Pstl and the fragments were ligated into the PstI site of pUC19 before transformation of Escherichia coli JM83. Out of thousands of transformants obtained by transformation of E coli JM83 with the genomic library, three plasmids were chosen. The sizes of the inserted DNAs were 2.9kb(2.4kb and 0.5kb) in pKTS1, 4.3kb in pKTS2 and 1.5kb in pKTS3, respectively. The DNA fragments used as probe KTS1(2.4kb), KTS2(4.3kb) and KTS3(1.5kb) were labeled digoxigenin-11-dUTP for the Southern hybridization. In Southern hybridization, all of the probes(KTS1, KTS2 and KTS3) reacted specifically to T sergenti DNA, but not to bovine leucocyte DNA. In order to find out the sensitivities of the digoxigenin-11-dUTP-labeled KTS1 and KTS3 as the probes, purified merozoite DNA and bovine DNA (control) were checked by dot blot hybridization with the probes. Both of the probes, KTS1 and KTS3, detected as minimum amount of 975pg of the T sergenti DNA, but not bovine DNA even to 500ng.

Detection of Antibodies Reacting with Anaplasma phagocytophilum and Ehrlichia chaffeensis from Cats, Horses and Cattle in Korea

채준석,허은정,박진호,최경성,양재혁,김도영,김준규,최귀철,강문일,J. Stephen Dumler,이성수,강태영 한국임상수의학회 2009 한국임상수의학회지 Vol.26 No.6

Antibodies to Anaplasma phagocytophilum and Ehrlichia chaffeensis were detected by the immunofluorescent antibody (IFA) test in sera collected from cats, thoroughbred horses and Holstein cattle in Gwangju, Jeonju and Jeju Island of Korea. Two hundred fifty four sera (33 feral and pet cats, 92 grazing horses and 129 grazing cattle) were obtained from Republic of Korea. Antibodies to A. phagocytophilum (titer ≥ 80) were detected in 6 of the 33 feral and pet cats (18.2%), and 1 seropositive cat (3.0%) also had antibodies to E. chaffeensis. Only 1 of 129 (0.8%) cattle and 2 of 92 (2.2%) horses had antibodies to A. phagocytophilum. Antibodies to E. chaffeensis were not detected in either of these animals. This is the first report of serological evidence of A. phagocytophilum and E. chaffeensis from cats, cattle and horses in Korea. These rickettsial agents could have an important impact on human health or impact animal health with economic losses among industrial grazing animals in Korea.

건설업 안전경영시스템모델 개발

채준석,갈원모,손기상 안전경영과학회 2001 안전경영과학회지 Vol.3 No.2

Safety management cost, US$ one thousand billion has been input to the construction area from 1995 to 2000 but the management quality is still in the developing country level. The above mentioned increase of accident rate are attributed to the generous code regulation, net fit to the sense, varying with time change, but the more fundamental reason of this is no practical construction safety management system, up to now. Hardware-orienting operation for the system could be a main reason of this problem. In this paper, we developed a model for efficient construction safety system. It was found from the case study that the model result in high efficient with low cost.

Identification and sequence analysis of small subunit ribosomal RNA gene of bovine Theileria isolates from Korea and Japan

채준석,박진호,권오덕,이주묵,Chae, Joon-seok,Park, Jin-ho,Kwon, Oh-deog,Waghela, Suryakant D.,Holman, Patricia J.,Wagner, Gerald G.,Lee, Joo-mook The Korean Society of Veterinary Science 1998 大韓獸醫學會誌 Vol.38 No.4

한국과 일본의 서로 다른 지역으로부터 소의 Theileria 분리주에 있어서 6가지 type(A부터 E 그리고 H)과 subtype(B1)의 small subunit ribosomal RNA(SSUrRNA) 유전자를 밝혔다. 이들 유전자 염기서열을 비교하여 본 바 염기서열의 위치 212~231, 261~270 그리고 632~690으로 3군데의 hypervariable region이 관찰되었다. SSUrRNA 유전자 염기서열 type A는 한국의 전북 분리주(KCB), 충남 분리주(KCN), 제주 분리주(KCJ)그리고 실험실 보관주(KLS)와 일본의 Shintoku 분리주(JHS)인 5개의 분리주에서 나타났으며, 이 염기서열은 Kenya의 Marula 분리주인 Theileria buffeli의 SSUrRNA 유전자(GenBank accession number Z15106)와 일치하였다. 한국의 경북 분리주(KKB)에서는 type B만이 관찰되었으나 그 외의 분리주에서는 2 type 이상의 유전자 염기서열이 관찰되었다. KCB와 JHS 분리주에서는 type A와 B, 강원 분리주(KKW)에서는 type B와 H, KCN 분리 주에서는 type A, C 및 D 그리고 KCJ 분리주에서는 type A, B, E 및 subtype B1이 관찰되었다. 한국과 일본 소의 Theileria 분리주에 있어서 여러 type의 SSUrRNA 유전자 염기서열이 나타나는 것으로 보아 혼함감염이 되어 있는 것으로 판단된다. Six different sequences types(A through E and H) and a subtype(Bl) of the small subunit ribosomal RNA(SSUrRNA) gene were found in bovine Theileria isolates from different areas of Korea and Japan. The sequences were aligned and three hypervariable regions were observed in the nucleotide position ranges 212~231, 261~270 and 632~690. Five of the Theileria isolates yielded sequence type A; these were the field isolates KCB, KCN, and KCJ, and the laboratory stock KLS, all from Korea, and a single isolate from Japan (JHS). This sequence type is identical to the SSUrRNA gene sequence listed for Theileria buffeli (GenBank Accession No. Z15106) from Marula, Kenya. The Korean field isolate KKB yielded only a single sequence type (B), but multiple sequence types were found in some isolates. For example, KCB and JHS isolates yielded both types A and B ; isolate KKW showed types B and H; isolate KCN showed types A, C, and D ; and isolate KCJ showed types A, B, E, and a subtype B1. Finding of the multiple sequences SSUrRNA gene sequences suggests that bovine Theileria isolates from both Korea and Japan may consist of mixed populations.

안전관리 기법에 관한 실증적 연구 : 건설작업을 중심으로

채준석,갈원모,손기상 대한안전경영과학회 2000 대한안전경영과학회지 Vol.2 No.1

Sourthern hybridization과 중합효소연쇄반응을 이용한 한국과 일본의 Theileria sergenti 비교

채준석,이주묵,권오덕,이승옥,채건상,Chae, Joon-seok,Lee, Joo-mook,Kwon, Oh-deog,Lee, Seung-ok,Chae, Keon-sang,Onuma, Misao 대한수의학회 1996 大韓獸醫學會誌 Vol.36 No.1

The T sergenti DNA fragments used as probes of KTS1(2.4kb) and KTS3(1.5kb) were labeled with digoxigenin-11-dUTP for the Southern hybridization. T sergenti DNAs from different geographic locations(Korea; Chonbuk, Kyungbuk, Chungnam, Kangwon, Cheju island, Japan; Shintoku, Shintoku 9209, Shintoku 9201, Shintoku 9202, Shintoku 9102) which had been digested with Pst I and EcoR I were probed by the digoxigenin-11-dUTP-labeled KTS1 and KTS3. As the results, the samples from Chonbuk, Kyungbuk, Cheju island in Korea and Shintoku, Shintoku 9209, Shintoku 9201, Shintoku 9102 in Japan were positively reacted, but the others from the other locations not reacted. In the comformation test of T sergenti DNA from different geographic locations, all of the samples were positively detected by PCR amplification.

Caine Babesia spp. 감염증예

채준석,인동철,한재철,김남수,이주묵,최인혁,Chae Joon-Seak,Ihn Dong-Chul,Han Jae-Chul,Kim Nam-Soo,Lee Joo-Muk,Choi In-Hyuk 한국임상수의학회 1989 한국임상수의학회지 Vol.6 No.1

A dog which was hospitalized to Veterinary Teaching Hospital, College of Veterinary Medicine, Jeonbug National University on December 28, 1988 was revealed severe anemia: hemoglobinuria and weakness. In the inspections, abdominal pain and spleno megaly at the ventral abdomen were detected by palpations. In the examinations of blood, the obtained results were summarized as follows: Babesla spp. was identified on the blood smear stained with Giemsa. The Babesia spp. was assumed to the Babesia gibsoni for the their small size and pleomorphism such as comma form, ring form and dot form. In the blood examinations of the patient, Ht: 22.5%, RBC:354${\times}$10$^4$/${\mu}\ell$, Hb: 8.8g/dl, serum protein: 8g/dl, and WBC count was 21, 425/${\mu}\ell$. In the chemical examinations of serum, the value of AST(GOT) was 30iu and ALT(GPT) was 20iu, respectively. The blood sugar was 60mg/d1. In the urine test, urine protein was 30mg/d1 and the hemoglobin In the urine was the +++ and occult blood reaction(Benzidine test) in the feces was +++. Splenomegaly was confirmed by X-ray examination. To confirm for the Babesia spp. infection, 5ml of the whole blood of the patient(3% of Parasitized erythrocytes) were inoculated into the cephalic vein of the two normal dogs. In the blood of experimental dogs which were inoculated parasitized blood, Babesia spp. was detected in the two doss and pleomorphic parasites were observed, too. In the blood examinations of No. 1 the Ht and RBC were decreased to 6.8% and 52${\times}$10$^4$/${\mu}\ell$, respectively. WBC count was 10.600/${\mu}\ell$ and serum protein was 6.8g/dl. The rates of parasitized erythrocytes were 15% in the experimental dog. Also +++ of the hemoglobin was detected in the urine. In the X-ray examination, splenomegaly was comfirmed and it was confirmed by autopsy of the experimental dog(No. 1).

건설안전관리의 발전 방향

채준석 한국안전학회(구-한국산업안전학회) 1993 `93 건설안전 학술 심포지움 Vol.- No.-