• 대한수의학회지
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• 제38권4호
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• pp.745-750
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• 1998

A sensitive and rapid enzyme immunoassay(EIA) to detect Escherichia coli O157 in ground beef was developed by using a sandwich type assay with polyclonal antibodies to E coli O157. E coli O157 in ground beef could be detected within 15hr, including incubation for 12hr in enrichment broth and 3hr in immunoassay. The EIA could detect $1.3{\times}10^5$ cells of E coli O157/g of ground beef without enrichment. The lowest limit of detection was 0.23 E coli O157 per g of meat after enrichment. Confirmation was required in the positive specimens in the EIA by culture method even though the negative specimens were not. These results suggested that the immunoassay could be a very efficient method for the screening E coli O157 in meat.

• 한국축산식품학회지
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• 제38권4호
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• pp.829-834
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• 2018

The objective of this study was to determine the minimum enrichment time for different types of food matrix (pork, beef, and fresh-cut lettuce) in an effort to improve Escherichia coli detection efficiency. Fresh pork (20 g), beef (20 g), and fresh-cut lettuce (20 g) were inoculated at 1, 2, and 3 Log CFU/g of Escherichia coli. Samples were enriched in filter bags for 3 or 5 h at $44.5^{\circ}C$, depending on sample type. E. coli cell counts in the samples were enriched in E. coli (EC) broth at 3 or 5 h. One milliliter of the enriched culture medium was used for DNA extraction, and PCR assays were performed using primers specific for uidA gene. To detect E. coli (uidA) in the samples, a 3-4 Log CFU/mL cell concentration was required. However, E. coli was detected at 1 Log CFU/g in fresh pork, beef, and fresh-cut lettuce after 5, 5, and 3-h enrichment, respectively. In conclusion, 5-h enrichment for fresh meats and 3-h enrichment for fresh-cut lettuce in EC broth at $44.5^{\circ}C$, and PCR analysis using uidA gene-specific primers were appropriate to detect E. coli rapidly in food samples.

• 한국미생물·생명공학회지
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• 제32권3호
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• pp.212-217
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• 2004

Glycosyl hydrolase family 26에 속하는 Bacillus subtilis WL-7 mannanase를 코드하는 유전자를 대장균에서 과잉 발현하였다. 아미노 말단의 signal peptide를 포함하거나 포함하지 않은 mannanase 유전자를 각각 pET24a(＋)에 도입하여 재조합 플라스미드 pETMAN과 pENS7를 제조하였다. 이들 플라스미드를 함유하는 Escherichia coli BL21(DE3)에서 mannanase를 발현시킨 결과 signal peptide가 제거된 mannanase유전자의 발현량이 매우 높았다. 그러나 배양온도 $37^{\circ}C$에서 pENS7를 함유한 재조합 대장균에서 과잉 발현된 mannanase는 대부분이 불활성 형태로 존재하였으며, 배양온도를 $31^{\circ}C$이하로 하였을 때 수용화 형태의 효소량이 증가하면서 효소활성이 높아졌다. IPTG에 의해 발현된 재조합 대장균의 균체파쇄 상등액 중에 존재하는 mannanase 활성은 배양온도 $25^{\circ}C$～28$^{\circ}C$에서 가장 높았으며, 전체 단백질량을 기준으로 볼 때는 배양온도 $31^{\circ}C$에서 비활성이 가장 높은 것으로 확인되었다.

• 미생물학회지
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• 제40권3호
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• pp.232-236
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• 2004

Streptomyces coelicolor A3(2)의 acetyl xylan esterase (AxeA)가 Escherichia coli의 과산화수소 저항성에 미치는 영향을 알아보고자 하였다. AxeA 발현은 isopropyl-$\beta$-thiogalactoside로 유도되었고 생산된 AxeA는 SDS-polyacrylamide gel electrophoresis방법으로 확인하였다. AxeA 발현에 따른 과산화수소 저항성의 변화를 E. coli의 생장곡선과 생존율을 통하여 조사하였다. AxeA가 발현되지 않으면 모든 처리 농도 (1 mM, 2.5mM, 5mM)에서 균의 사멸이 일어났다. AxeA가 발현되는 조건에서는 5mM을 제외한 과산화수소 1mM와 2.5mM에서 E. coli의 사멸이 저지되었다. 또한 1.5mM의 과산화수소에 대한생존율이 59%에서 74%로 높다졌다. 동시에 E. coli의 최고생장온도에서에 근접한 $45^{\circ}C$에서의 생존율도 증가되는 결과를 얻었다. 그러므로 AxeA 단백질은 산화적 스트레스와 온도스트레스에 대해 교차 저항성을 나타내는 역할을 한다고 결론지었다.

• 미생물학회지
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• 제34권4호
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• pp.231-235
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• 1998

Escherichia coli 세포에서 intramolecular triplex가 형성될 수 있는지를 조사하기 위하여 genetic assay를 도입하였다. Two dimensional gel electrophoresis 방법을 사용하여 $(G)_9AATTC(G)_9$ 혹은 $(GAA)_9TTC(GAA)_8$ sequences가 in vitro에서 intramolecular triplex 구조를 형성하는 것을 확인하였다. Genetic assay를 위하여 temperature-sensitive EcoRI methylase 유전자를 포함한 플라스미드와 triplex를 형성할 수 있는 $(G)_9AATTC(G)_9$ 혹은 $(GAA)_9TTC(GAA)_8$ sequences를 포함한 플라스미드로 E. coli를 cotransform 하였다. EcoRI methylase가 발현되는 permissive temperature에서 pur pyr sequences를 포함하는 플라스미드는 EcoRI methylase 작용에 kinetically 저항성을 보여주었다. 이러한 결과는 pur pyr sequence들이 EcoRI methylase가 작용하기 어려운 triplex 구조를 E. coli 세포에서 형성한다는 것을 증거한다.

• 한국축산식품학회지
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• 제37권6호
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• pp.799-803
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• 2017

In the present study, centrifugation and filtration pretreatments were evaluated to decrease sample preparation time and to improve the sensitivity and specificity of multiplex polymerase chain reaction (PCR) for the detection of low levels of pathogenic Escherichia coli in various foods. Pathogenic E. coli (E. coli NCCP11142, E. coli NCCP14037, E. coli NCCP 14038, E. coli NCCP14039, and E. coli NCCP15661) was inoculated into pork, beef, and baby leafy vegetables at 1, 2, and 3 Log CFU/g. The samples were shaken 30 times (control), then centrifuged or filtered. DNA extracts from the samples were subjected to PCR using the $Powerchek^{TM}$ Diarrheal E. coli 8-plex Detection Kit. In the pork samples, no E. coli was detected in the control samples, while E. coli were detected in 100% of 3-Log CFU/g inoculated and centrifuged samples, and in 100% of 2 and 3-Log CFU/g inoculated, and filtered samples. In the beef samples, all control samples appeared to be E. coli-negative, while E. coli was detected in 50-75% of centrifuged samples, regardless of inoculated level, and in 100% of 2 and 3-Log CFU/g inoculated, and filtered samples. In baby leafy vegetables, E. coli were not detected in 25-50% of the control samples, while E. coli were detected in 0-25% of the centrifuged samples, and 75-100% of the filtered samples, depending on the inoculum amount. In conclusion, filtration pretreatment can be used to minimize sample preparation time, and improve the sensitivity and specificity of rapid detection of pathogenic E. coli in various foods.

• 한국수의병리학회:학술대회논문집
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• 한국수의병리학회 2003년도 추계학술대회초록집
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• pp.30-30
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• 2003

Escherichia coli strains associated with diarrhea have been divided into the following six major categories on the basis of pathogenic mechanisms: enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), enteropathogenic E. coli (EPEC), enterohemorrhagic E. coli (EHEC), enteroaggregative E. coli (EAggEC) and diffusely adherent E. coli (DAEC).$\^$15,18/ EPEC, EAggEC, and DAEC strains were classified by their ability to produce distinct patterns of adherence to cultured epithelial cells in virto: localized (LA), aggregative (AA), and diffuse (DA) adherence. The objective of this study was to investigate the relationship of the adherence patterns with AIDA-positive E. coli isolated from diarrheic pigs. (omitted)

• Fisheries and Aquatic Sciences
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• 제18권3호
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• pp.235-240
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• 2015

Seasonal variation in the prevalence of norovirus in oysters Crassostrea gigas was investigated and compared to levels of Escherichia coli, a fecal indicator in oysters. Oysters were collected from Iwon-myeon, Taean-gun, Korea, a primary production area for European Union export of oysters between 2013 and 2014. We observed seasonality in the prevalence of norovirus in oysters, with a higher prevalence and viral load detected during winter months. Oysters taken from production areas that complied with the European Union standard for raw consumption (< 230 MPN/100 g of E. coli) had 22.1% of the samples test positive for norovirus (15/68 samples). However, norovirus was not detected in any of the samples (0%, 0/4 samples) that were collected from production sites that exceeded the standard fecal contamination level for raw consumption (> 230 MPN/100 g of E. coli). These results indicated that there is a negative correlation between the prevalence of norovirus and high levels of E. coli in oysters. Therefore, our results suggest that current food safety guidelines using only a bacterial fecal contamination indicator, E. coli, may not adequately assess shellfish production areas for viral and bacterial contamination.

• 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.

• 대한의생명과학회지
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• 제10권3호
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• pp.293-298
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• 2004

This study was performed to compare the growth rates, cellular fatty acid compositions and morphology by using electron microscope of Escherichia coli (E. coli) grown in various conditions including different concentrations of phenol, CdCl₂ and HgCl₂. Ninety eight E. coli strains were isolated from Naktong river and human feces. The content of unsaturated fatty acids, especially 16:1ω7c and 18:1ωc increased as the concentration of phenol and CdCl₂ increased. The content of unsaturated fatty acid increased up to 50 ppb of HgCl₂, but decreased at 75 ppb of HgCl₂. There were more unsaturated fatty acids than saturated fatty acid in the presence of toxic substances. However, the ration was reversed when the affected E. coli was transferred to toxic substance free fresh trypticase soy broth medium. Also, by using transmission electron microscope these cells were observed to various morphological deformation by heavy metals and their deposition on the surface. From these results, we suggested that the changes of major fatty acids composition and morphology of E. coli may be considered to indicate contaminated levels of heavy metals or organic solvents. The information presented here may be useful in predicting effects of heavy-metal and organic solvent contamination in streams and provides a basis for further studies of metal or organic solvent effects on microbial communities.