• 동아시아식생활학회지
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• 제11권1호
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• pp.26-32
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• 2001

독소원성 대장균(enterotoxigenic Escherichia coli, ETEC, EC81, serotype O148:H28)이 생산하는 heat labile enterotoxin(LT)를 검색해 본 결과, reversed passive latex agglutination(RPLA)법에 있어서는 2배로 희석한 용액 (50 ng)으로부터 64로 희석한 용액 (1.56 ng)에서까지 양성반응을 보였으며 polymerase chain reaction (PCR)기법에 있어서는 10 ng으로부터 1 pg희석용액에서까지 147-base pair(bp)의 LT DNA fragment가 확인되었다. Clostridium perfringens A형 (NCTC8238, Hobbs serotype 2)이 생산한는 장독소를 검색해 본 결과, RPLA법에 있어서는 2배로 희석한 용액 (50 ng)으로부터 64로 희석한 용액 (1.56 ng)에서 까지 양성반응을 보여 독소원성대장균이 생산하는 LT와 일치하였으나, PCR기법에 있어서는 10ng로부터 10 pg 희석용액에서 까지 354-bp의 DNA fragment가 확인되어 독소원성대장균이 생산하는 LT보다 1/10의 낮은 감도를 보였다. PCR기법은 RPLA법에 비하여 훨씬 신속하고 소량의 sample로 장독소를 확인할 수 있었다.

• Journal of Plant Biotechnology
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• 제29권4호
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• pp.287-293
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• 2002

병독성 대장균 (K88ac)의 pilin gene을 분리하여 이를 당근에 도입한 후 이의 발현을 유도하여 자돈 설사병 예방 및 치료를 위한 당근경구백신 개발을 목적으로 하였다. 형질전환을 통해 494 세포주를 확립하였고, western분석을 통하여 0.1~4 $\mu\textrm{g}$/g의 pilin 단백질 발현을 확인하였으며 백신식물 생산에 적합한 세포주 2종 (M1-17, Y14-1)을 선발하여 포장생산 및 임상실험에 적용하였다. 쥐를 대상으로 당근 경구 투여시 병원균 항체 생성 여부와 면역 유도를 위한 최적 농도 지표를 파악하기 위하여 1주 간격으로 형질전환 당근 1g, 3g, 5g을 경구투여 한 결과 백신 당근 3g 투여시 10$\mu\textrm{g}$의 재조합 pilin 백신을 경구 투여한 것에 비해 다소 높은 항체 생성을 나타냈으며 3g의 분량이 쥐 면역 유도를 위한 적정량으로 확인하였다. 자돈에 대한 백신당근 경구 투여시 자돈 설사병 보호 효과를 정량적으로 구명하기 위하여 분석한 자돈의 일당 증체량은 형질전환 당근 투여시 평균 60g 이상 더 높은 증체량을 보였으며, 질병방제 효과를 구명하기 위하여 장독성 병원균을 인위 접종한 결과 대조구에서만 fecal score 3의 심각한 설사를 확인하였다. 본 연구를 통해 개발된 당근백신은 효율적 투여방법 등에 대한 후속 실험을 통하여 산업적 이용 가능성이 탐색될 예정이다.

• 한국식품위생안전성학회지
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• 제24권1호
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• pp.19-26
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• 2009

본 연구는 집단급식 안전성 확보 및 식중독 방지를 위해 단체급식 안전성에 관하여 전문가를 대상으로 국내 집단급식 식중독 원인을 조사하고, 미래 집단급식 식중독 발생을 예측하였다. 델파이 설문 결과 국내 집단급식 식중독 원인은 '식재료 위생관리 미흡', '손세척', '구역구분', '유통업체 보관' 등이 가장 크게 영향을 주는 것으로 나타났다. 미래 집단급식 식중독 발생 예측 조사 결과 Vibrio parahaemolyticus, Escherichia coli(EPEC), non-typhoid Salmonella serotypes, Staphylococcus aureus, Escherichia coli(ETEC), Norovirus, hepatitis A virus가 지속적으로 식중독을 발생시킬 것으로 예측하였으며, 집단급식에서 식중독 유발 가능성이 높은 식재료로는 영유아(1-6세), 초등학생(7-12세), 중 고등학생(13-19세)의 식재료 섭취량과 식중독 유발 가능성을 고려해 볼 때 '과일류', '우유', '생선류', '돼지고기', '계란', '쇠고기'였으며, 식단 메뉴로는 '비빔밥', '콩나물무침', '시금치나물', '오이생채', '잡채', '돼지불고기'가 위해도가 높을 것으로 예측되었다. 집단급식소에서 사용빈도가 높을 것으로 예측된 가공식품은 '냉동, 냉장보관식품' 이었으며, 살균방법은 '가열처리', '화학적 살균소독제'로 나타났다. 저장법은 '냉장($10^{\circ}C$ 이하)', '냉동($-20^{\circ}C$ 이하)'법이, 배식형태는 '집단급식소에서 직접 배식' 형태가 집단급식소에서 향후 사용빈도가 높을 것으로 예측되었다. 본 설문조사 결과는 집단급식 전 과정의 체계적인 위생관리시스템의 도입과 식중독예방 식단 구성을 위한 기초자료로 활용될 것으로 사료된다.

• 한국동물위생학회지
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• 제42권2호
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• pp.93-112
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• 2019

Calf diarrhea is a common disease in young claves and is still a major cause of productivity and economic loss in livestock farms. Fecal samples from Korean native cattle (n=100) with diarrhea from 64 farms in Gwangju area, Korea from september 2017 to December 2018 were examined for shedding of important protozoan parasitic, viral and bacterial pathogens using culture, rapid test kit and PCR methods. Of 57 (89.1%) of the 64 Korean native cattle farms examined had samples infected with at least one of the investigated pathogens. Among 100 fecal samples, 88 samples were positive for at least one the twelve pathogens and 51 samples were simultaneously positive for two or more pathogens by culture and PCR assay. Bovine group A rotavirus (BRV) was the most common pathogen, found in 43/100 (43.0%) samples on 32/64 (50.0%) farms. Subsequently, kobuvirus (30.0%), pathogenic E. coli (29.0%), bovine parvovirus (17.0%), Giardia spp. (13.0%), Eimeria spp. (10.0%), Clostridium perfringens type A (8.0%), bovine torovirus (8.0%), bovine viral diarrhea virus (6.0%), bovine coronavirus (5.0%), bovine norovirus (2.0%) and Cryptosporidium spp. (2.0%) were detected. Nebovirus, kırklareli virus, bovine adenovirus, Salmonella spp. and intestinal parasites were not detected. Of the 72 calves sampled in this age group, 64 (88.9%) samples were positive for at least one enteropathogen. BRV was identified in 34/72 (47.2%) samples from 27/48 (56.3%) farms. Subsequently, pathogenic E. coli (30.6%), kobuvirus (29.2%), BPaV (22.2%), Giardia spp. (15.3%), Eimeria spp. (9.7%), BVDV (6.9%), Cl. perfringens type A (6.9%), BCoV (4.6%) and Cryptosporidium spp. (2.8%) were detected in fecal samples. A total of ninety-six strains of E. coli were isolated from one hundred fecal samples collected from Korean native cattle with diarrhea. The presence of stx1, stx2, eaeA, LT, STa, STb, ehxA, saa, F4, F5(K99), F6, F17, F18 and F41 genes in the isolates was investigated by PCR. Out of ninety-six E. coli isolates screened for specific genes, 30 strains E. coli were identified to harbor shiga toxin-producing E. coli (STEC) 7 (7.3%), enterohemorrhagic E. coli (EHEC) 8 (8.3%), enteropathogenic E. coli (EPEC) 6 (6.3%), enterotoxigenic E. coli (ETEC) 2 (2.1%) and STEC/ETEC hybrid 7 (7.3%). This study provides epidemiological estimates of the prevalence of Korean native cattle's enteropathogens in Gwangju area, Korea, which would be used for cattle farmers and veterinarians to select appropriate therapeutic method.

• Journal of Animal Science and Technology
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• 제62권4호
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• pp.543-552
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• 2020

For efficient prevention and treatment of enteric colibacillosis, understanding about latest virulence factors and antimicrobial resistance of Escherichia coli is essentially needed. The aim of this study was to survey antimicrobial resistance and determine the prevalence of fimbriae and enterotoxin genes among 118 pathogenic E. coli isolates obtained from Korean pigs with diarrhea between 2016 and 2017. The genes for the toxins and adhesins were amplified by polymerase chain reaction (PCR). The susceptibility of the E. coli isolates to antimicrobials were tested using the standard Kirby-Bauer disk diffusion method. The most prevalent fimbrial antigen was F18 (40.7%), followed by F4 (16.9%), and the most prevalent combinations of toxin genes were Stx2e (21.2%), STb:EAST-1 (19.5%), and STa:STb (16.9%), respectively. Among the pathotypes, enterotoxigenic E. coli (ETEC) was the most predominant (67.8%), followed by Shiga-toxin producing E. coli (STEC, 23.7%). We confirmed high resistance rates to chloramphenicol (88.1%), tetracycline (86.4%), streptomycin (86.4%), and ampicillin (86.4%). And the majorities of isolates (90.7%) showed multi-drug resistance which means having resistance to 3 or more subclasses of antimicrobials. Results of this study can be a source of valuable data for investigating the epidemiology of and control measures for enteric colibacillosis in Korean piggeries.

• Journal of Magnetics
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• 제20권3호
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• pp.284-289
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• 2015

This paper discusses general methods of modelling magnetic saturation in steady-state, two-axis (d & q) frame models of dual stator induction generators (DSIG). In particular, the important role of the magnetic coupling between the d-q axes (cross-magnetizing phenomenon) is demonstrated, with and without cross-saturation. For that purpose, two distinct models of DSIGs, with and without cross-saturation, are specified. These two models are verified by an application that is sensitive to the presence of cross-saturation, to prove the validity of these final methods and the equivalence between all developed models. Advantages of some of the models over the existing ones and their applicability are discussed. In addition, an alternative is given to evaluate all saturation factors (static and dynamic) by just calculating the static magnetizing inductance which is simply the magnitude of the ratio of the magnetizing flux to the current. The comparison between the simulation results of the proposed model with experimental results gives a good correspondence, especially at startup.

• 한국미생물학회:학술대회논문집
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• 한국미생물학회 2000년도 International Meeting 2000
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• pp.58-65
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• 2000

Diarrheal diseases are a major cause of both illness and death in developing countries and are caused by rotavirus, Shigella spp., Salmonella spp., enterotoxigenic Escherichia coli (ETEC), and Vibrio spp. In this study, for the development of vaccine against diarrheal diseases caused by Shigella sonei, Salmonella typhimurium, E. coli O157, and Vibrio cholerae, cloning and nucleotide sequence analysis of genes and characteristics of their gene products in E. coli were performed. For construction of attenuated strain of S. sonnei KNIH104 and Salmonella typhimurium KNIH100, the aroA genes were cloned, respectively. The recombinant plasmid $_pJP{\Delta}A45$ containing aroA deleted region and suicide vector $(_pJP5603)$ was constructed. The aroA gene deleted mutants were constructed using this recombinant plasmid. For cloning gene encoding antigenic region of E. coli O157 KNIH317, the O-antigen synthesis gene cluster and sit gene was cloned. The E. coli XL1-Blue cells harboring this recombinant plasmid showed cytotoxicity in Vero cells. The ctx gene was cloned for tile purpose of antigenic region against V. cholerae KNIH002. Sequence analysis confirmed that the virulence gene cassette was consisted of ace, zot, ctxA and ctxB genes.

• 한국미생물·생명공학회지
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• 제48권2호
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• pp.121-128
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• 2020

Diarrhea is a major public health concern associated with pathogenic Escherichia coli infections. Food-borne pathogenic E. coli can lead to large diarrheal outbreaks and hence, there is a need to estimate the frequency of pathogenic E. coli load in the various types of meat available in markets. In the present study, we classified and characterized diarrheagenic E. coli isolates collected from 399 raw meat samples from retail sources in Korea. Shiga toxin-producing E. coli (STEC) were detected in 11 (9.7%) samples, including nine strains (8.0%) in beef and two strains (1.8%) in chicken. The frequency of the detected virulence markers were as follows: astA, 28.3%; escV,18.6%; eaeA,17.7%; ent, 7.0%; EHEC-hly, 4.4%; stx1, 3.5%; and stx2, 3.5%. We did not observe any typical EPEC, EIEC, or ETEC virulence determinants in any of the samples. The STEC serotype O26 was detected in one sample, but no other serogroups (O91, O103, O128, O157, O145, O111, and O121) were found. Further research is needed to better understand the virulence mechanism of STEC serotypes, their ecology, and prevalence in animals, food, and the environment. These results will help improve risk assessment and predict the sources of food poisoning outbreaks.

• Journal of Dairy Science and Biotechnology
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• 제39권2호
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• pp.51-62
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• 2021

Escherichia coli are the predominant facultative bacteria found in the gastrointestinal tract of animals and humans. Some strains of E. coli that acquire virulence factors and cause foodborne and waterborne diseases in humans are called pathogenic E. coli and can be divided into five pathotypes according to the virulence mechanism: EAEC, EHEC, EIEC, EPEC, and ETEC. Although selective media have been developed to detect E. coli, distinguishing pathogenic strains from non-pathogenic ones is difficult because of their similar biochemical properties. Therefore, it is very important to find a new and effective diagnostic method to identify pathogenic E. coli. With recent advances in molecular biology and whole genome sequencing, the use of polymerase chain reaction (PCR) is increasing rapidly. In this review paper, we provide an overview of pathogenic E. coli and present a review on PCR detection methods that can be used to diagnose pathogenic E. coli. In addition, the possibility of real-time PCR incorporating IAC is introduced. Consequently, this review paper will contribute to solving the current challenges related to the detection of pathogenic E. coli.

• 한국환경보건학회지
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• 제42권3호
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• pp.189-195
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• 2016

Objectives: To ensure the microbiological safety of groundwater, it was confirmed whether waterborne pathogenic bacteria in groundwater samples tested positive for total coliforms in the Chungcheongnam-do Province region. Methods: Total colony counts, total coliforms and fecal coliforms were tested according to the process mandated by the drinking water quality testing standards of Korea. DNA was extracted from the samples, tested positive for total coliforms, and then subjected to real-time PCR to detect waterborne pathogenic bacteria. Results: A total of 115 samples were inadequate for drinking water. Thirty-one cases (27%) showed positive for fecal coliforms and nine cases (7.8%) showed total colony counts exceeding drinking water standards. Twenty-seven cases (23.5%) showed three items (total colony counts, total coliforms and fecal coliforms). Using the real-time PCR method, waterborne pathogens were detected in 57 cases (49.6%) in 115 samples. Seventy-eight cases of waterborne pathogenic bacteria were detected (including duplications): 27 cases of pathogenic E. coli (EPEC (19), ETEC (5), EHEC (1), EAEC (1) and EIEC (1)); 45 of Bacillus cereus; two of Yersinia spp.; two of Salmonella spp.; one of Staphylococcus aureus; one of Clostridium perfringens. Conclusion: The real-time PCR method can offer rapid and accurate detection of waterborne pathogenic bacteria. Therefore, this assay could be an alternative to conventional culture methods and can further ensure the microbiological safety of groundwater.