• Journal of Food Hygiene and Safety
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• v.31 no.6
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• pp.393-398
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• 2016

The objective of this study was to isolate pathogenic Escherichia coli from fresh vegetables with selective media and Petrifilm, and identify a suspicious colony using biochemical identification. Twenty gram of lettuce, twenty gram of cabbage and ten gram of sprout were prepared, and a 5-strain mixture of pathogenic E. coli (Enterohemorrhagic E. coli NCCP11142, Enterotoxigenic E. coli NCCP14037, Enteropathogenic E. coli NCCP14038, Enteroaggregative E. coli NCCP14039, Enteropathogenic E. coli NCCP15661) was inoculated to obtain 1, 2 and 3 log CFU/g. Eighty to ninety milliliter of buffered peptone water (BPW) was placed and pummeled for 60 s. As a results, the Petrifilm method was all positive, but enrichment method of qualitative analysis was negative except for 3-log CFU/g inoculated lettuce. Regarding biochemical identification of pathogenic E. coli, the identification rates were dependent on type of methods and vegetables; lettuce: API 20E 100% (44/44), Microgen GNA 100% (44/44) and Food System 66.7% (10/15), cabbage: API 20E 64.7% (22/34), Microgen GNA 50% (16/32) and Food System 60% (9/15), sprout: API 20E 65.1% (28/43), Microgen GNA 62.3% (27/43) and Food System 53.3% (8/15). These results could be useful in determining an appropriate method to detect pathogenic E. coli in fresh vegetables.

• Korean Journal of Microbiology
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• v.32 no.1
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• pp.47-52
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• 1994

The cells of Campylobacter jejuni heat-shocked at 48${\circ}C$ for 30 min synthesized the heat shock proteins of HSP90, HSP66 and HSP60. Those heat shock proteins were found to correspond to the heat shock proteins of HSP87, HSP66 (DnaK), and HSP58 (GroEL) of E. coli, respectively. By Southern blot analysis of the chromosomal DNAs of C. jejuni with groESL and dnaK genes of E. coli as DNA probes, the heat shock genes of C. jejuni which are homologous to the E. coli groESL and dnaK genes were found to exist in the chromosomal DNA. The genomic libraries of C. jejuni were constructed with the cosmid vector pWE15 and the groEL gene of C. jejuni were cloned in E. coli B178 groEL44 temperature senstive mutant. The hybrid plasmid (pLC1) was inserted with the DNA fragment (about 5.7kb in size) containing the groEL gene. E. coli groEL44 mutant cell transformed with the pLC1 could grow at 42${\circ}C$ by synthesizing the HSP60 of C. jejuni and regained the susceptibility to the ${\lambda}$ vir phage by expression of the groEL gene in the cloned cells. These indicated that the groEL products of C. jejuni had chaperon effects by synthesizing the heat shock proteins in the cloned cells of E. coli.

• Food Science of Animal Resources
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• v.38 no.4
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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.

• Korean Journal of Veterinary Service
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• v.43 no.2
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• pp.53-58
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• 2020

Enterovirulent Escherichia coli are among the most important causes of diarrhea in cattles. Between January and December, 2017, a total of 150 stool specimens from cattles were investigated for enterohaemorrhagic E. coli (EHEC), enteropathogenic E. coli (EPEC), enteroaggregative E. coli (EAEC), enterotoxigenic E. coli (ETEC) and enteroinvasive E. coli (EIEC) using real-time PCR. 131 E. coli were isolated from feces. The most frequently isolated pathotype in feces was EHEC (37 isolates). EPEC, ETEC and EAEC were detected in feces with 14, 7 and 3 respectively. EIEC was not detected. Antimicrobial resistance test was performed by agar disc diffusion method with 14 antimicrobials. Enterovirulent E. coli isolates showed the highest antimicrobial resistance to ampicillin 61.3%, followed by tetracycline 54.5% and streptomycin 45.5%. They had low resistance to amikacin 11.4%. Of 44 isolates, 37 (84.1%) were resistant to more than 2 antimicrobials. futher study a highest antimicrobial susceptibility to trimethoprim/sulfamethoxazole 50.0% and florofenicol 47.7%.

• Korean Journal of Clinical Laboratory Science
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• v.47 no.2
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• pp.71-77
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• 2015

Antimicrobial resistant bacteria has recently emerged and been disseminated in livestock environments because of excessive use of antimicrobial agents for the therapeutic and growth promotion purposes to food animals. In particular, there is potential for multidrug-resistant bacteria that can be transmitted from animals to mankind. In this study, we investigated the genotypes of E. coli strains isolated from humans and chickens using multi-locus sequence typing (MLST) and antimicrobial resistance patterns by disk diffusion method along with integron study involving antimicrobial resistance mechanisms. From July 2013 to July 2014, E. coli strains isolated from clinical specimens (n=44) and poultry chickens (n=34). ST131 (n=20) was most common in human-derived E. coli. ST752 (n=7) was most common in chicken-derived E. coli, with four isolates each for ST117, ST189, and ST69. Of the 44 E. coli strains isolated from humans, 25 of had a class 1 integron, as opposed to only 11 of 34 strains in the E. coli isolated from chickens. There were differences in genotypes and antimicrobial resistance patterns between the chicken-derived and the human-derived E. coli.

• Journal of Microbiology and Biotechnology
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• v.27 no.12
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• pp.2141-2150
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• 2017

This study was conducted to develop a screening method using Colilert-18 and a device for the detection of E. coli from agri-food production environments and fresh vegetables. The specificity and sensitivity of Colilert-18 by temperature ($37^{\circ}C$ and $44^{\circ}C$) were evaluated with 38 E. coli and 78 non-E. coli strains. The false-positive rate was 3.8% (3/78) and 0% (0/78) at $37^{\circ}C$ and $44^{\circ}C$, respectively. The detection limit of E. coli at $37^{\circ}C$ at <1.0 log CFU/250 ml was lower than that at $44^{\circ}C$. The efficiency of the developed device, which comprised an incubator equipped with a UV lamp to detect E. coli in the field, was evaluated by measuring the temperature and UV lamp brightness. The difference between the set temperature and actual temperature of the developed device was about $1.0^{\circ}C$. When applying the developed method and device to various samples, including utensils, gloves, irrigation water, seeds, and vegetables, there were no differences in detection rates of E. coli compared with the Korean Food Code method. For sanitary disposal of culture samples after experiments, the sterilization effect of sodium dichloroisocyanurate (NaDCC) tablets was assessed for use as a substitute for an autoclave. The addition of one tablet of NaDCC per 50 ml was sufficient to kill E. coli cultured in Colilert-18. These results show that the developed protocol and device can efficiently detect E. coli from agri-food production environments and vegetables.

• The Korean Journal of Food And Nutrition
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• v.11 no.4
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• pp.416-419
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• 1998

Identification of escherchia coli K1 polysaccharide antigen isolated from urine specimens of urinary tract infections in children were performed from of 1992 to 1993 in Kyoto, Japan. The serotypes of E. coli were categorized that O1:H7, O2:H6, O2:H7, O16:H6, O18:H7, O18:H￣, and O135:H44 among 14 strains isolated from urine specimens of urinary tract infections in children by the serological test. And, one strain (O18:H￣, isolation rate: 7.1%) of E. coli K1 polysaccharide antigen among 14 strains were isolated from urine specimens of urinary tract infections in children by the bacteriophage test.

• Journal of Microbiology and Biotechnology
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• v.9 no.6
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• pp.722-725
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• 1999

Filamentation-suppressed recombinant Escherichia coli strain harboring the Alcaligenes latus polyhydroxyalkanoate (PHA) biosynthesis genes and the E. coli ftsZ gene was constructed and cultivated for the production of poly(3-hydroxybutyrate) [P(3HB)] with high concentration and high content. By the pH-stat fed-batch culture of this recombinant E. coli strain XL1-Blue(pJC5), the final cell concentration and P(3HB) concentration obtained in 44.25h were 172.2g cell dry weight/l and 141.9g P(3HB)/l, respectively, resulting in productivity of 3.21g P(3HB)/l-h. More importantly, the P(3HB) content obtained was 82.4 wt %, which was significantly higher than that obtained with the recombinant E. coli harboring only the PHA biosynthesis genes.

• Korean Journal of Microbiology
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• v.44 no.2
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• pp.93-97
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• 2008

RraA is a recently discovered protein inhibitor that regulates the enzymatic activity of RNase E, which plays a major role in the decay and processing of RNAs in Escherichia coli. It has also been shown to regulate the activity of RNase ES, a functional Streptomyces coelicolor ortholog of RNase E, which has 36% identity to the amino-terminal region of RNase E. There are two open reading frames in S. coelicolor genome that can potentially encode proteins having more than 35.4% similarity to the amino acid sequence of RraA. DNA fragment encoding one of these RraA orthologs, designated as RraAS2 here, was amplified and cloned in to E. coli vector to test whether it has ability to regulate RNase E activity in E. coli cells. Co-expression of RraAS2 partially rescued E. coli cells over-producing RNase E from growth arrest, although not as efficiently as RraA, induced by the increased ribonucleolytic activity in the cells. The copy number of ColEl-type plasmid in these cells was also decreased by 14% compared to that in cells over-producing RNase E only, indicating the ability of RraAS2 to inhibit RNase E action on RNA I. We observed that the expression level of RraAS2 was lower than that of RraA by 4.2 folds under the same culture condition, suggesting that because of inefficient expression of RraAS2 in E. coli cells, co-expression of RraAS2 was not efficiently able to inhibit RNase E activity to the level for proper processing and decay of all RNA species that is required to restore normal cellular growth to the cells over-producing RNase E.

• Korean Journal of Veterinary Research
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• v.48 no.3
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• pp.295-303
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• 2008

One hundred and sixty nine Escherichia (E.) coli strains isolated from fecal samples of aquatic birds in Geumho river basin and Dalseong park were tested by agar dilution method to determine their susceptibility patterns to 14 antimicrobial agents. The distribution of tetracycline resistance determinants (tetA, tetB, tetC, tetD and tetE) were also examined by PCR in 76 tetracycline-resistant ($TC^r$) E. coli isolates. The high resistance was observed in tetracycline, cephalothin and ampicillin (45.0~36.7%). Resistance of E. coli isolates derived from Dalseong park to tetracycline, cephalothin, ampicillin and streptomycin (65.7~44.8%) were significantly higher than those isolated from Geumho river basin (31.4~14.7%). About seventy percent (70.4%) of the strains isolated were resistant to one or more drugs tested. Thirty (39.5%) of 76 $TC^r$ E. coli isolates which were resistant to one or more drugs transferred all or a part of their resistance patterns to the recipient strain of E.coli J53 by conjugation. All of $TC^r$ E. coli isolates contained at least one or more of 5 tet genes examined. The most common genes found in these isolates were tetA (60.6%) and followed by tetB (7.9%) and tetC (1.3%). However, tetD and tetE were not found in any of the isolates tested. Twenty one (27.6%) of $TC^r$ E. coli isolates had two determinants, tetA/tetB (20 strains), tetA/tetC (1 strain). And two strains (2.6%) contained three determinants (tetA/tetB/tetC).