• 대한수의학회지
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• 제36권2호
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• pp.371-378
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• 1996

Three and 2 strains of E coli O157 were isolated from fecal materials of cattle (390) and pigs (420) in Korea, respectively. One strain of O157:H7 isolated from cattle and 2 strains of O157:H7 isolated from pigs were identified as verotoxin-1 (VT-1) produing strains and 2 strains (O157:H7 and O157:H-) isolated from cattle were identified as verotoxin-2 (VT-2) producing strains by neutralization test on HeLa and Vero cells. Culture supernatants of the isolates were cytotoxic to HeLa and Vero cells. The levels of cytotoxin produced by isolates were $10^2{\sim}10^4$ cytotoxic dose($CD_{50}$)/ml. Also, VT-2-converting bacteriophage was isolated from KSC109 strain which had been isolated from cattle. Molecular weight of the phage DNA was determined as approximately 45 Kb in 0.8% agarose gel electrophoresis, and morphology of the phage stained with phosphotungstic acid was observed by transmissible electron microscopy.

• 대한수의학회지
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• 제40권3호
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• pp.525-531
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• 2000

Verotoxin을 산생하지 않은 Escherichia coli O157과 verotoxin을 산생하는 E coli(VTEC)를 건강한 소의 분변에서 분리하여 생화학적 및 유전적인 특성에 대해서 비교하였다. E coli O157 : nonH7(운동성은 있으나 H혈청형이 7이 아님)의 sorbitol 분해능과 ${\beta}-glucurondase$ 활성은 E coli O157 : H7이 나타내는 것과는 차이가 있었다. 그리고 uidA 유전자는 verotoxin 산생능과 상관없이 sorbitol과 ${\beta}-glucuronidase$ 음성인 E coli O157 : H7에서 특이적으로 검출되었다. 한편 6개 목장에서 수거한 소 분변 45예에서 VTEC를 분리한 결과, 7주(15.6%)가 분리되었으며 이들은 모두 sorbitol을 분해하였으며 ${\beta}-glucurondase$ 활성이 있었으나 장벽 부착인자를 지배하는 eaeA 유전자가 없었다. 비록 소가 VTEC의 보균원으로 추정되나, 정상 소에서 분리한 VTEC는 eaeA 유전자가 결여된 균주가 많으므로 공중위생학상 eaeA 유전자를 보유한 E coli 보다 위해성이 낮으며, 이러한 결과는 왜 사람에서 유행하는 VTEC 혈청형과 소에서 유행하는 것과 차이가 있는지를 일부 설명해준다.

• 대한수의학회지
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• 제36권2호
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• pp.379-387
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• 1996

The objects of the present study were to establish the method of purification, subunit dissociation of verotoxin-2 (VT2) produced by Escherichia coli O157:H7, and to investigate the characteristics of purified verotoxin-2 such as molecular weight and composition of amino acid. The results were summerized as follows; Verotoxin-2 was extracted by addition of polymyxin B sulfate into bacterial cell lysate prepared from Escherichia coli O157:H7(KSC109). As an initial step, the bacterial cell lysate was precipitated with 30% saturated ammonium sulfate. The precipitated crude toxin was then subjected to anion-exchange, chromatofocusing and cation-exchange chromatography. Using this scheme, we obtained highly purified toxin with a specific activity of $1.1{\times}10^9$ $CD_{50}/mg$. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE) for purified VT2 showed two protein bands. The upper band, approximately 32 Kd, was supposed as A subunit and the lower band, approximately 7.7 Kd, was supposed as B subunit. When the toxin was separated in the subunit-dissociating solution, two peaks emerged with retention times of 15 and 28 min by HPLC. These peaks represented A subunit and B subunit, respectively. The amino acid composition of purified VT2 were made up in order of glutamic acid, histamine, asparaginic acid, histidine, lysine, alanine and leucine etc. The largest amount among the amino acid composing VT2 was methionine.

• Asian-Australasian Journal of Animal Sciences
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• 제20권5호
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• pp.789-794
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• 2007

A PCR-aided monitoring of verotoxin-producing Escherichia coli (VTEC) was performed over the period of 12 months by using fresh feces collected monthly from 5 dairy cows that had been identified as VTEC carriers. The PCR products were confirmed to be verotoxin genes by Southern hybridization using a gene fragment of verotoxin 2 as a probe. Although seasonal variation of VTEC shedding seemed to depend on each cow, several factors may have influenced the frequency of detection. Shedding of VTEC tended to be reduced during grazing from the middle of May up to the beginning of October. Only one cow was positive for VTEC in August. Dry-off was also suggested to have a depressive effect on VTEC shedding, i.e. 3 of 4 dry cows showed no shedding of VTEC. Contrary to these factors, winter or indoor rearing tended to increase VTEC with only 5/24 samples being negative during the period from November to April. Total VFA concentration was higher (p<0.05) in VTEC-positive feces than in VTEC-negative feces, while fecal pH and VFA proportions were not different. Partial sequences of verotoxin genes from feces of 4 VTEC-positive cows were nearly identical (99-100%), suggesting that gut bacteria sharing the same gene were distributed among the cows. The present results indicate that grazing and dry-off could be factors which reduce VTEC shedding, while winter/indoor rearing may be a factor which increases the shedding, possibly through on-farm interactions.

• Journal of Microbiology
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• 제38권2호
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• pp.93-98
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• 2000

Nine hundred patients diagnosed with diarrhea or hemorrhagic uremic syndrome in the Kyungpook Province, Korea, were examined from November 1998 to February 2000. One patient in Kumi appeared to possess the Escherichia coli O157:H7 strain, which is very important in clinical decision making and public health action. The isolated strain, an E. coli O157:H7 KM, contained a 60 MDa plasmid and typical virulence genes including the verotoxin 2 gene, ehxA gene (encoding enterohemorrhagic hemolysin), and eae (encoding attaching and effacing protein-intimin) gene. This strain produced only verotoxin 2. Pulsed field gel electrophoretic analysis showed that the genomic organization of the E. coli O157:H7 KM strain may differ greatly from those of representative strains previously reported in the United States and Japan.

• 대한수의학회지
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• 제50권3호
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• pp.231-237
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• 2010

This study was conducted to investigate characteristics and antimicrobial susceptibility of Escherichia (E.) coli isolates isolated from vaginal mucosa and clitorial fossa of 105 Thoroughbred mares suspicious of the genital disease in Korea during the period from March 2006 to July 2007. Ninety six E. coli isolates were identified as standard biochemical properties and using BIOLOG system. Fifty three isolates (55.2%) could be classified into a total of 21 O serotypes and forty three isolates (44.8%) were non-typeable with 51 O antisera used in this study. The verotoxin 1 (VT 1) and verotoxin 2 genes were analyzed by multiplex PCR. Among them, one isolate was detected VT 1 gene (130 bp). Most of isolates showed a high susceptibility in ciprofloxacin (100%), enrofloxacin (100%), norfloxacin (100%), cefoxitin (96.9%), gentamicin (96.9%), sulphamethoxazole (96.9%), nitrofurantoin (94.8%), amikacin (93.8%), nalidixic acid (92.7%) and tetracycline (90.6%). These results may provide the basic information to establish strategies for the treatment and prevention of reproductive disease in Thoroughbred mares in Korea.

• 한국미생물·생명공학회지
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• 제50권3호
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• pp.387-394
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• 2022

Verotoxin-1 (VT-1) or Shiga-like toxin 1 (Stx-1) is produced by enterohemorrhagic Escherichia coli (EHEC) and is an AB5 holotoxin with a strong inhibitor of protein synthesis. VT-1 is a type 2 ribosome-inactivating protein (RIP) that has been shown to have cytotoxic and anticancer potential by inducing necrosis, apoptosis, and cell cycle arrest, making it a promising antitumor candidate. Here, we tested the cytotoxicity of VT-1 on CaCo2 and NCM425 cell lines and the results showed that VT-1 was more potent on CaCo2. Morphological changes were also evaluated on the cellular level and the results showed that VT-1 caused a decrease in viable cell count, altered cell membrane permeability, and an increase in total nuclear intensity. On the other hand, VT-1 displayed a lesser impact on mitochondrial membrane potential (MMP) and cytochrome c release. On the expression of caspases 3 and 9, VT-1 exhibited an insignificant effect on both which alongside the mitochondrial membrane potential (MMP) and cytochrome c results, might indicate that CaCo2 suffered from the necrosis process as a mechanism of cell death after exposure to VT-1.

• 한국식품위생안전성학회지
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• 제15권3호
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• pp.241-247
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• 2000

국내 유통식품에서 verotoxin(VT)을 생성하는 대장균의 오염도를 조사하고 이로 인한 식중독의 발생을 사전에 예방하기 위하여 1997년부터 1999년 사이 햄버거. 식육 및 채소류에서 VT생성 대장균의 분포조사를 실시하였으며, 이 결과 분리된 대장균에 대한 분자생물학적 특성을 조사하였다 식육, 햄버거 및 채소류 총 1,700건의 식품 중에서 VT를 생성하는 대장균이 검출된 것은 3건으로 매우 낮은 검출율을 보였는데, 분리주의 혈청형은 각각 026 : H4, 0157 : H7 및 055 : Hl2였다. 혈청형 026 : H4 분리주는 VT I과 VT II를 생성하였고,055 : Hl2주는 VT I을 각각 생성하였으며, 두 분리주에서 eae유전자와 60 MDa plasmid DNA는 검출되지 않았다. 혈청형 0157 : H7주는 VT II를 생성하였고 60 MDa plasmid DNA는 검출되었으나 eae유전자는 확인되지 않았다. 따라서 분리된 VT생성 대장균들은 eae 유전자가 검출되지 않아 질병유발 가능성은 낮은 것으로 추정되었다. 항생제 내성능에서 혈청형 0157 : H7 분리주는 ampicillin과 streptomycin에 내성을 나타낸 반면, 혈청형 026 : H4주와 혈청형 055 : Hl2주는 ampicillin, carbenicillin, cephalothin, trimethoprim/sulfamethoxazole과 tetracycline에 내성을 보여 분리주의 다재내성능을 확인하였다 세포배양액으로 실시한 vero cell과 HeLa cell에 대한 세포독성능은 3주의 분리주에서 확인되었다.

• 대한수의학회지
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• 제33권4호
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• pp.773-779
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• 1993

• 한국동물위생학회지
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• 제28권4호
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• pp.407-412
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• 2005

Ten isolates of Verotoxin-producing Escherichia coli (VTEC) were detected in slaughtered cattle and investigated their phenotypic characteristics and antimicrobial susceptibility. None of the isolates was positive for eae gene. Only one isolate was positive for uidA gene. Eight out of ten isolates of VTEC were originated from broker's cattle. Thus microbiological monitoring for broker farms should be performed to minimize VTEC contamination. In the antimicrobial susceptibility test, all the isolates were highly resistant to bacitracin and lincomycin whilst they are susceptible to apramycin and neomycin.