• Journal of Microbiology
• /
• 제40권2호
• /
• pp.91-97
• /
• 2002

Enteroaggregative E. coil (EAEC) is an important aethiological causal agent of diarrhea in people of developed and undeveloped countries. Different in vitro and in vivo models have been proposed to study the pathdgenic and immune mechanisms of EAEC infaction. The aim of this study was to analyze whether BALB/c mice could be used as an animal model to study EAEC pathogenesis Six-week-old BALB/c mice were inoculated with EAEC strain 042 (044:H88) nalidixic acid resistant, and re-inoc-ulated ten days after. Mice feces were monitored for the presence of the EAEC strain over a period of 20 days . Bacteria were enumerated on MacConkey agar containing 100$\mu$g of nalidixic acid per ml. Results showed that 35% of the animals were colonized for 3 days, 15% for 5 and 10% for more than 7 days . After re-inoculation only 16% of the animals remained colonized for more than 3 days. During the necropsy, the intestinal fluid of same of the infected animals presented mucus and blood. Six of these fluids showed the presence of IgA antibodies againset Pet toxin and IgG natibodies raised against the toxin were also detected in the animal serum. Histopathologic evidence confirms the stimulation of mucus hypersecretion, an increased amount of goblet cells and the presence of bacterial aggregates in the apical surfaces of intestinal epithelial cells. Edema was present in the submucosa. These results suggest that BALB/c mice could be used as an animal model for in vivo study of EAEC infection.

• 대한미생물학회지
• /
• 제22권2호
• /
• pp.139-145
• /
• 1987

Adherence to HEp-2 cells has been proposed as a virulence characteristic of enteropathogenic E. coli (EPEC). The role of the HEp-2 adherent E. coli was evaluated in a group of children with endemic diarrhea admitted to Hanyang University Hospital in Seoul, Korea. HEp-2-adherent E. coli was detected in fecal samples of 59 (59%) of 100 cases and ten (22.7%) of 44 concurrent control children (p<0.0005). Adherence was exhibited by 15 serogroups and subgroups, but within these groups more than one adherence pattern was frequently observed. Of 17 strains belonging to traditional infantile EPEC serogroups, 12(70.6%) gave a positive adherence. Of 45 enterotoxin producing strains, 24 (53.3%) gave a positive adherence. HEp-2-adherent strains that did not belong to classic EPEC serogroups and did not produce heat-stable and/or heat-labile enterotoxins(referred as enteroadherent E. coli, EAEC) was found in 29 (29%) of the patients with diarrhea and in six (13.6%) of the well children (p<0.05). From 22 of the 29 cases, no pathogen other than EAEC was isolated. These findings strongly implicate EAEC as the cause of diarrhea in the children. Our study supports the concept that EAEC may be an important cause of endemic diarrhea in Korean children.

• Journal of Microbiology and Biotechnology
• /
• 제24권3호
• /
• pp.421-426
• /
• 2014

To characterize the extended-spectrum beta-lactamases (ESBLs) in diarrheagenic Escherichia coli from Korea in 2008-2011, we screened seven enterotoxigenic E. coli (ETEC) and one enteroaggregative E. coli (EAEC) that produce ESBLs from a nationwide survey. All eight isolates produced CTX-M-type ESBLs, including CTX-M-12 (n = 4), CTX-M-14 (n = 2), and CTX-M-15 (n = 2). PCR-based replicon typing indicated that the $bla_{CTX-M-12}$ genes of four ETEC isolates were carried on a conjugative IncF plasmid, whereas the $bla_{CTX-M-14}$ of one EAEC was located on an IncK plasmid. This is the first report of the occurrence of $bla_{CTX-M}$ genes in clinical isolates of EAEC in Korea. The ESBL-producing isolates were shown to be different based on pulsed-field gel electrophoresis and multilocus sequence typing, whereas the four isolates with CTX-M-12 were clonally related. These observations raise an alarm for the spread of plasmid-mediated resistance to ESBL among diarrheagenic E. coli.

• Journal of Microbiology and Biotechnology
• /
• 제24권6호
• /
• pp.862-868
• /
• 2014

Certain Escherichia coli (E. coli) strains have the ability to cause diarrheal disease. Five types of diarrheagenic E. coli have been identified, including EHEC, ETEC, EPEC, EAEC, and EIEC. To detect these five diarrheagenic types rapidly, we developed a one-step multiplex PCR (MP-PCR) assay using nine primer pairs to amplify nine virulence genes specific to the different virotypes, with each group being represented (i.e., stx1 and stx2 for EHEC, lt, sth, and stp for ETEC, eaeA and bfpA for EPEC, aggR for EAEC, and ipaH for EIEC). The PCR primers were constructed using MultAlin. The sensitivity and specificity of the constructed multiplex PCR primers were measured using DNA isolated from diarrheagenic E. coli strains representing each group. The limits of detection were as follows: $5{\times}10^1CFU/ml$ for EHEC, $5{\times}10^3CFU/ml$ for ETEC expressing lt and sth, $5{\times}10^4CFU/ml$ for ETEC expressing stp, $5{\times}10^2CFU/ml$ for EPEC, $5{\times}10^4CFU/ml$ for EAEC, and $5{\times}10^2CFU/ml$ for EIEC. To confirm the specificity, C. jejuni, C. perfringens, S. Typhimurium, V. parahaemolyticus, L. monocytogenes, Y. enterocolitica, B. cereus, and S. aureus were used as negative controls, and no amplification was obtained for these. Moreover, this kit was validated using 100 fecal samples from patients with diarrhea and 150 diarrheagenic E. coli strains isolated in Korea. In conclusion, the multiplex PCR assay developed in this study is very useful for the rapid and specific detection of five diarrheagenic E. coli types. This single-step assay will be useful as a rapid and economical method, as it reduces the cost and time required for the identification of diarrheagenic E. coli.

• 대한미생물학회지
• /
• 제22권2호
• /
• pp.155-162
• /
• 1987

To determine the prevalence of antibiotic resistance in fecal E. coli and to investigate possible associations between antibiotic resistance and other plasmid-mediated virulence properties, antibiotic disk susceptibility tests for nine antibiotics were done on 141 strains of E. coli isolated from diarrheal children and well controls. Eighty two percent of the test strains were resistant to one or more antibiotics. Antibiotics to which the test strains were most resistant in descending order were ampicillin (85%), trimethoprim/sulfamethoxazol (60%), and cephalothin (55%). Seventy nine percent of these resistant strains were resistant to two or more antibiotics. All 141 test strains were sorted into enterotoxigenic E. coli (ETEC), enteropathogenic E. coli (EPEC), enteroadherent E. coli (EAEC) and non-pathogenic E. coli and the percentages of strains resistant to multiple antibiotics were compared. Among ETEC regardless of its source, multiple drug resistance was more frequent in strains producing heatstable enterotoxin (ST) only than in strains producing only heat-labile enterotoxin (LT) or both. In EAEC, multiple resistance was more frequently associated with strains isolated from diarrheal patients than with those from well controls. The major antibiotic resistance patterns possessed by multiple resistant enteropathogenic strains were $SXT^R$ $AM^R$, $CR^R$, and $SXT^R$ $AM^R$ $CR^R$. Of 28 ST- producing $SXT^R$ ETEC, 26(96%) were also resistant to ampicillin and 17 (61%) were resistant to cephalothin. The similar pattern was observed in EAEC and EPEC as well. This study has important implications for the treatment of E. coli diarrhea with antibiotics because it is possible that dissemination of virulence could occur under the force of selective antibiotic pressure. In addition, this study suggests that the in vivo efficacy of SXT in treating diarrheal illness be reevaluated.

• 한국동물위생학회지
• /
• 제43권2호
• /
• pp.53-58
• /
• 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%.

• 한국식품과학회지
• /
• 제47권1호
• /
• pp.126-131
• /
• 2015

본 연구에서는 한국의 전통적인 식품인 비빔밥에서 2005년에서 2011년에 분리한 대장균에 대한 특성을 연구하였다. 전체 1,142건의 비빔밥에서 84 균주(7.4%)의 대장균이 분리되었다. 분리한 균주의 항생제 내성 분석 및 다제 내성 패턴은 ampicillin의 내성이 가장 높았으며 6 균주(7.2%)에서는 다제 내성을 보였다. 분리한 균주의 병원성 유전자 13종의 유무를 확인한 결과 5 균주(5.9%)에서 ent 유전자를 보유하여 STEC/EPEC 형의 대장균으로 분류되었다. 그러나 EIEC, EAEC, ETEC 형은 검출되지 않았다. 병원성 유전자가 검출되지 않은 대장균에 대한 혈청형은 O103 (1.2%), O91 (1.2%), O128 (6.0%)으로 확인되었고 O26, O157, O145, O111, O121 혈청형은 없었다. 동일한 O 혈청형 균주는 독소 유전자를 가지고 있는 것과 없는 것이 동시에 있다고 보고 되어 있고 혈청형으로 병원성 유전자를 구분하는 것은 균주의 특성에 따라 차이가 많이 나므로 향후 연구가 필요하다. 위해 요소가 있는 균주에 대해서 유전적 상동성을 시험한 결과 균주의 상동성이 낮아서 서로 다른 오염원에서 분리 되었음을 유추 할 수 있었다. 비빔밥은 다양한 재료를 사용하며 제조 과정이 복잡하기 때문에 대장균에 의한 오염의 가능성이 매우 높은 식품이다. 그러므로 원재료부터 완성까지 위생적으로 철저하게 조리해야 하며 교차 오염 방지 및 관리를 통해 안심하고 섭취할 수 있도록 해야 한다.

• 한국식품위생안전성학회지
• /
• 제32권2호
• /
• pp.129-134
• /
• 2017

병원성 대장균군은 전세계적으로 식중독을 발생하는 중요한 병원균으로 알려져 있다. 국내 유통중인 생닭과 조리된 닭가공품 356시료에서 대장균 80균주(22.5%)를 분리하여 병원성 유전자 multiplex PCR를 이용하여 분석한 결과 astA 유전자가 26균주(32.5%)에서 검출되었으며, escV 유전자는 17균주(21.3%) eaeA 유전자는 16균주(20.0%) 검출되었다. 특히 식품에서 문제가 되는 장출혈성 대장균 유전자 stx 1는 3균주(3.8%)와 stx 2, EHEC-hly는 각 1균주(1.3%)에서 검출되었다. 병원성 유전자를 토대로 STEC, EPEC, EHEC, EIEC or EAEC의 병원성 대장균으로 분류한 결과 45균주(56.3%)가 검출되었으며 typical EPEC, EIEC and ETEC의 병원성 대장균은 검출되지 않았다. STEC 병원성 대장균은 O152, O1, O116, O26, O25, O119, O153 혈청형이 분리되었다. 특히 생닭에서 병원성 대장균에 대한 검출률이 높으므로 생닭을 가공 조리 시 교차 오염이 발생하기 않도록 조리 도구, 조리 환경에 대한 철저한 관리가 필요하다고 생각된다.

• 한국환경보건학회지
• /
• 제45권5호
• /
• pp.434-442
• /
• 2019

Objectives: This study was performed in order to report the epidemiological features of a food poisoning outbreak caused by multiple pathogens in a high school in Chungcheongnam-do Province, Korea in April 2019 and to suggest measures to prevent a similar incidence. Methods: A total of 39 patients with diarrhea were examined. Environmental samples were obtained from 6 food handlers, 4 food utensils, 72 preserved foods served during the food poisoning outbreak, 9 door handles, 10 drinking water samples from water dispensers, and 6 ground water samples from water taps. These analyzed to detect viruses and bacteria. Results: Among the 39 patients, 21 cases (53.8%) of enteroaggregative E. coli (EAEC), 7 cases of Staphylococcus aureus (17.9%), and 17 cases of norovirus (43.6%) were positive, and in 16 of the cases a co-infection with at least one other pathogen were observed. EAEC was assumed to be transmitted from contaminated drinking water because it was also detected in the water sample from a water dispenser in the dormitory. Staphylococcus aureus was isolated only in the fecal samples of patients, meaning it was not possible to trace its origin. The genotype of norovirus detected in the drinking water and ground water was consistent with that isolated from patients, and it was determined that the norovirus infection originated from the school's water environment. Conclusions: These findings indicate that a lack of environmental hygiene management related to school meals caused the food poisoning incident. In particular, a lack of management of drinking water, water supply, and personal hygiene should be pointed out. This should be urgently addressed and continuous monitoring should be carried out in the future. In addition, students and staff should be educated and trained to improve their personal hygiene.

• Journal of Dairy Science and Biotechnology
• /
• 제28권2호
• /
• pp.7-11
• /
• 2010

In recent times, antimicrobial resistance has been a concern because of its relation to national health and food safety. In this study, we reviewed the management of antibiotics and antimicrobial-resistant microorganisms in domestic and foreign countries and analyzed microorganisms and antimicrobial resistance in dairy farms and raw milk. The isolates from dairy farms were Micrococcus spp., Staphylococcus chromogens, Escherichia coli, Bacillus licheniformis, Escherichia coli O157:H7, Pseudomonas plecoglossicida, Enterobacter spp., and Kluyvera intermedia. Rummeliibacillus stabekisii, Paenibacillus badius, Pantoea agglomerans, B. oleronius, B. fusiformis, and B. badius were isolated from feed E. coli and Kurthia gibsonii, from feces and S. pasteuri, S. aureus, S. chromogenes, and Salmonella spp., from raw milk. Pathogens isolated from dairy farms and raw milk were tested for susceptibility to 20 types of antibiotics. E. coli (EAEC) and E. coli O157:H7 (EHEC) isolated from dairy farms, E. coli (EAEC) isolated from feces, and Salmonella spp. isolated from raw milk showed resistance to multiple antibiotics. These results show that antimicrobial-resistant microorganisms should be more effectively managed to improve the safety of dairy farms.