목 적: 세균성 각막염 치료에 주로 사용되는 퀴놀론계 항균제의 감수성 저하로 나타난 Serratia marcescens 균주에 대해 내성기전에 관여하는 주요 target인 gyrA와 parC 유전자를 분석하여 감수성 저하의 원인을 파악하고자 한다. 방 법: 퀴놀론계 항균제 감수성 저하 Serratia marcescens 균주들을 대상으로 genomic DNA를 추출 하여 PCR를 통해 증폭한 후 gyrA와 parC 유전자 QRDR의 DNA sequencing을 분석하였다. 결 과: parC 유전자 분석에서 SM5 균주는 Tyr51→His, Thr59→Asp으로 이중 아미노산 변이가 나타났 으며, SM6 균주는 Thr59→Asp으로, SM13 균주는 Tyr51→His으로 단일 아미노산 변이가 나타났다. 그리고 3 균주 모두 gyrA와 parC 유전자에 다수의 silent mutation이 관찰되었다. 결 론: target 유전자의 변이로 인한 항균제의 감수성이 저하되고 있다는 것을 확인할 수 있었으며, 특히 parC 유전자가 퀴놀론계 항균제의 감수성 저하를 일으키는 주요 표적임을 확인할 수 있었다. 앞으로도 세균성 각막염 치료에 퀴놀론계 항균제의 사용이 계속될 것이며, 많은 내성 균주들이 발생할거라 예측된다. 따라서 항균제를 사용할 때 내성에 대한 주의가 반드시 필요할 것으로 사료된다.

적리를 동반하는 돼지 분변에서 용혈성 균주를 분리 동정한 후, 장 독소 생성 유 무 및 항생제 감수성 시험을 수행하였다. 분리된 용혈성 균주 B. cereus BY06의 gyrB 유전자 염기서열을 분석한 결과, B. cereus와 99% 유사성을 나타내었다. PCR법에 의한 장 독소 유전자 검출 시험에서 B. cereus BY06는 장 독소 분비 양성으로 판정됨에 따라 설사형 식중독 균임이 확인되었다. B. cereus BY06를 이용한 항균제의 감수성 시험 결과, penicillin G에는 내성을 나타낸 반면 cephalothin, vancomycin, clindamycin, fusidic acid, gentamicin, ciprofloxacin, tetracycline 및 rifampin에 대하여 감수성을 나타냈다. 본 연구를 통해 돼지 분변에서 분리된 B. cereus 균주는 설사를 유발하는 장 독소를 분비하며, penicillin G에 대한 내성을 확인하였다.

본 연구는 항균성 코팅제의 개념과 동향파악으로 항균성 코팅제의 연구개발의 방향을 설정하는데 도움을 주기 위한 것이다. 항균제는 미생물을 제거하거나 성장을 저지하는데 사용되는 화합물이며 항균 코팅제에 함유되는 항균제용 재료는 무기물, 금속, 저분자 유기물, 천연유기물, 고분자가 있다. 항균코팅제는 생활용품, 병원용품, 산업용품, 전자제품, 의류, 건축 내장재 등의 표면의 항균성 부여에 쓰인다. 기존 항생제는 세균의 세포벽을 손상하지 않고 미생물을 침투하나 항균성 고분자는 세포막을 파괴하므로 항생제의 내성을 방지할 수 있다. 대부분의 고분자 항균제는 양이온 고분자에 초점을 맞추고 있다. 항균제의 분자설계와 코팅제 배합의 합리화로 항균제의 선택성, 내구성, 독성 문제가 개선될 것이다.

This study was carried out to investigate efficacy of aerosol sanitizer with natural antimicrobial and organic acids against Escherichia coli O157:H7, Salmonella Typhimurium and Listeria monocytogenes. The artificially inoculated pathogens on stainless steel coupon were treated with grapefruit seed extract (GFE), acetic acid,citric acid and lactic acid in model cabinet for 5 min. The number of three foodborne pathogens with individual treatment was reduced by 0.34-3.77 log units, treatment with GEF + organic acid was reduced by 1.72-3.89 log units and treatment with GEF + organic acid + alcohol was reduced by 1.46-5.05 log units. By treatment with GEF + lactic acid + alcohol in scale-up model system for 10 min. Populations of E. coli O157:H7, S. Typhimurium and L. monocytogenes were reduced by 3.42, 2.72 and 2.30 log units from the untreated control respectively. From the above result,aerosol sanitizer with natural antimicrobial agents and organic acid can be used as an environmental sanitation method with satisfying the consumer demand on safe food.

This study was carried out to investigate antibiotic resistances of 181 isolates in Staphylococcus aureus isolated from 450 specimens which were a General Hospital personnel in Cheonan province. Of the 181 isolates, 89 isolates (49.2%) were methicillin resistant and 92 isolates were susceptible. They were resistant to penicillin (80.1%), clindamycin (18.8%), oxacillin (49.2%), cefepime (28.2%), cefotetan (34.3%), ampicillin (66.3%), gentamicin (27.6%), tetracycline (17.7%) and erythromycine (43.1%), also. In resistant rates of the 89 MRSA, 5 antibiotics were more than 50%, but in those of the 92 MSSA, only 1 antibiotics was. Also they showed very highly multi-drug resistance patterns to 15 antibiotics as follow : One hundred and forty five isolates(82.3%) showed antibiotic resistance to more than one kind of the 15 kinds antibiotics. In the multiple resistant patterns, one drug resistant isolates were 18 isolates (9.9%), 2 drugs 29 (16.0%), 3 drugs 21 (11.6%), 4 drugs 29 (16.0%), and more than 5 drugs were 52 isolates (28.7%). The number of more than 5 drugs resistant strains in MSSA were only 1 strain, but those in MRSA were 51 isolates (57.3%). Forty five isolates (24.9%) produced enterotoxin serotype C and all MRSA.

This study was carried out to investigate the antimicrobial resistance pattern and distribution of resistance gene determinants in fecal E. coli from chicken. Antimicrobial susceptibility test was performed on a total of 109 fecal E. coli isolates from chicken, collected in Gyeonggi, Chungbuk, Jeonnam and Jeonbuk province from March to November 2003, by the disk agar diffusion method. Eighteen commonly used antimicrobial agents approved in Korea as veterinary medicine were tested: ampicillin (AM), amoxicillin/clavulanic acid (AMC), cephalothin (CF), cefozolin (CZ), cefoxitin (FOX), cefotaxime (CTX), cefepime (FEP), imipenem (IPM), streptomycin (S), gentamicin (GM), amikacin (AN), ciprofloxacin (CIP), enrofloxacin (ENO), norfloxacin (NOR), trimethoprim/sulfamethoxazole (SXT), erythromycin (E), chloramphenicol (C) and tetracycline (TE). Higher resistance rates (≥50%) were observed against 9 antimicrobial agents including AM, CF, S, CIP, ENO, NOR, SXT, E and TE. Resistance was most frequent for TE in 105 E. coli isolates (96.3%). Twenty-two isolates (20.2%) of the isolates showed multiple antimicrobial resistance to 8, and 19 isolates (17.4%) showed to 7 antimicrobial agents. The distribution of the resistance gene determinants for S and TE was assessed by PCR in resistant isolates. Thirty isolates possessed the strA, strB, and aadA gene, 25 isolates possessed the strA and strB gene among the 66 streptomycin-resistant isolates. Fifty one isolates possessed only the tetA gene, 22 isolates possessed the tetA and tetB gene, 11 isolates possessed only the tetB gene among the tetracycline-resistant isolates.

Antifungal activity of seven natural antimicrobials, such as ε-polylysine, extract of Yucca shidigera, TLS (vitamin B1 derivative), BMB-FS, chitooligosaccharide, KDSP 001, and KDSP 002 were investigated for their applications in functional beverage. Five fungi including Aspergillus niger, Penicillium citrinum, Rhizopus oryzae, Fusarium moniliforme, and Mucor rouxii were applied as test fungi and mininum inhibitory concentrations (MICs) of antimicrobials were examined. TLS exhibited the strongst antifungal activity among tested antimicrobials and the growth of all fungi was inibited at 100 ppm. The antifungal activity of BMB-FS appeared different. The growth of Fusarium moniliforme was inhibited by BMB-FS at 100 ppm, but it exhibit antifungal activity on P. citrinum, and M. rouxii at 1000ppm. MICs of TLS appeared to be 60 ppm for four test fungi except A. niger (100 ppm). These results indicate the possible usage of TLS and BMB-FS as natural antimicrobials in functional beverage.

The purpose of this study was to investigate the antimicrobial activities of natural antimicrobials (10 formulas, F1~F10) against yeasts in functional beverages. The growth rates of yeasts were different with the ten different natural antimicrobial formulas tested. Yeasts grew for 14 days and the antimicrobial effect was observed between 14 and 18 days. Levels of S. cerevisiae, Z. bailii, and P. membranaefaciens were reduced to the limit of detection (ND) < 10 CFU/mL) after 28 days. Resistance against the antimicrobial effect was greatest for P. membranaefaciens, which grew to a level of 0.12~1.48 log10 CFU/mL after 14 days and was reduced to a level of 1.61~3.55 log10 CFU/mL after 28 days. The resistance of C. albicans was also high with a growth level of 0.13~1.28 log10 CFU/mL after 14 days and reduction to 1.51~5.30 log10 CFU/mL after 28 days. The antimicrobial effect of F10 was strongest for P. membranaefaciens. Every treatment reduced the microbial levels to 2.68~5.62 log10 CFU/mL after 6 months. F2, F4, F5, F6, and F10 reduced the C. albicans level to ND after 28 days while F1, F3, F8, and F9 reduced yeasts to the ND level after 6 months. The antimicrobial activities observed here will be useful for development of natural antimicrobials.

This study was conducted to evaluate the MSPD and HPLC method about simultaneous determination for residual synthetic antimicrobials of sixteen species such as sulfonamide etc. in livestock products. Elution solvent used in HPLC was ethylacetate : acetonitrile (4 :1), and mobile phases for solvent A and B were water : methanol : acetonit rile : phosphric acid (700:250:50:0.2) and 100% acetonitrile respectively. The detector and absorbency used in HPLC was UV 266 nm. This study showed the reduction effect of 99.1% for organic solvents, 94% for experimental steps, 95% for analytical time and manpower and 98.9% for costs compared with korea food standard method. The average recovery rates for chicken, bovine, pork and milk were 67.7% 96.2 %, 67.7%-96.6%, 70.0%-96.2%, and 13.8%-97.8%.

In order to investigate the species, serobiological characteristics and antibiotic sensitivity of Pseudomonas spp, we isolated Pseudomonas spp from 57 spring waters around Seoul area for spring, summer and autumn and identified Pseudomonas spp by biochemical characteristics and serological method. And also we tested the antibiotic sensitivity test by discdiffusion method. Of 57 spring waters tested, Pseudomonas spp were isolated from 33 spring waters(57.9%). Isolation rate of Pseudomonas spp in spring season was 28.1%, summer 21.1% and autumn 28.1%. Only 1 spring water was detected Pseudomonas spp in all seasons and 9 (15. 8%) were detected for 2 seasons and 13 (22.8%) were for only 1 season. Isolation rate of Pseudomonas spp at Mt. Cheonggye was 50% and followed by Mt. Bookhan 35.7%, Mt. Daemo 33.3%, Mt. Dobong 29.6%, Mt. Surak 25.9%, Mt. Woomyun 22.2% and Mt. Bulam 7.4%. Of 44 Pseudomonas spp, 22 strains (50%) were identified by Ps. putida, Ps. aeruginosa, Ps. tluorescens and Ps. mendocina were identified 6 strains (13.6%), respectively. 4 strains (9.1%) were identified by Ps. aureofaciens. Of 6 Ps. aieruginosa, serotype A was 2 strains, B, E, G, and K was 1 strain, respectively. Of 44 Pseudomonas spp, resistance rate to amoxicillin was 90.9% and followed by chloramphenicol 84.1%, tetracycline 84.1%, carbenicillin 81.8%, nalidixic acid 68.2%, neomycin 38.6%, streptomycin 31.8%, gentamicin 4.6%, kanamycin 4.6% and colistin 2.3%. Ps. aseruginosa was more sensitive to carbenicillin than other Pseudomonas spp isolated from spring waters in Seoul area but more resistant to kanamycin, and Ps. aureofaciens was no resistant to streptomycin. Among multiple drug resistance, resistance to 5 drugs was 31.8%, 4 drugs 15.9%, 7 drugs 13.6%, 1 drug and 2 drugs 4.6%, and 8 drugs 2.3%, respectively. The multiple resistance patterns detected highestly were NA-CB-C-TE-AMC (18.2%), NA-CB-N-C-TE-AMC (13.6%), CB-C-TE-AMC (11.4%) and NA-CB-N-C-TE-AMC-S (9.1%).

The antibacterial and antifungal effect of grapefruit seed extract(GFSE) was investigated for its purpose of application to a diverse spectrum of field as sanitizers, disinfectants and preservatives. GFSE showed comparatively high content of such flavoniods as naringin and hesperidin and ascorbic acid. GFSE containing a low level of organic acids is a heavy viscous and water-soluble liquid. As a result of the antimicrobial test of GFSE, Bacillus subtilis and Aspergillus oryzae did not survive at detectable levels when treated with more than 100 ppm of GFSE. The minimum inhibitory concentrations of GFSE for a wide variety of pathogenic and putrefactive bacteria, fungi and yeasts were 100 ppm and 250 ppm, respectively. In the comparable electron micrograph of microbial cells treated with GFSE or not, we could conclude that GFSE destroy microorganisms by disrupting the functions of the cell wall membrane and microbial spores.