• Korean Journal of Veterinary Research
• /
• v.36 no.2
• /
• pp.371-378
• /
• 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.

• Korean Journal of Veterinary Research
• /
• v.40 no.3
• /
• pp.525-531
• /
• 2000

Verotoxin non-producing E coli O157 strains have been isolated from cattle feces and compared in particular regard to biochemical properties and genotypes with verotoxin-producing E coli (VTEC). E coli O157 : nonH7 strains had different phenotypes in sorbitol fermentation and ${\beta}-glucuronidase$ activity from E coli O157 : H7. Regardless of verotoxin production ability of E coli O157 : H7, uidA gene was uniquely detected from sorbitol and ${\beta}-glucuronidase$ negative E coli O157 : H7. Forty five fecal samples from 6 dairy farms were obtained and VTEC was detected as 15.6% (7 strains) of the samples. Most VTEC isolates were positive for sorbitol fermentation and ${\beta}-glucuronidase$ activity but negative for eaeA gene. This study suggested that cattle could be a reservior for VTEC. However, absence of eaeA gene in VTEC isolates from most of healthy cattle suggested that they might be less virulent than eaeA-positive E coli against human health.

• Korean Journal of Veterinary Research
• /
• v.36 no.2
• /
• pp.379-387
• /
• 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
• /
• v.20 no.5
• /
• pp.789-794
• /
• 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
• /
• v.38 no.2
• /
• pp.93-98
• /
• 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.

• Korean Journal of Veterinary Research
• /
• v.50 no.3
• /
• pp.231-237
• /
• 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.

• Microbiology and Biotechnology Letters
• /
• v.50 no.3
• /
• pp.387-394
• /
• 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.

• Journal of Food Hygiene and Safety
• /
• v.15 no.3
• /
• pp.241-247
• /
• 2000

The incidence of verotoxin-producing Escherichia coli(VTEC) was surveyed in domestic foods including hamburger, raw meats and vegetables from 1997 to 1999. The molecular biological characteristics of the isolates were analyzed. Three VTEC strain were isolated from 1,700 samples. Serotypes of those isolates were 0157 : H7, 026 H4, and 056 : Hl2, respectively. Serotype O26 : H4 produced VT I and VT II, and 055 Hl2 isolate produced VT I, however the 60 MDa plasmid DNA and eae gene were not found from both strains. One 0157 : H7 isolate produced VT II and harbour 60 MDa plasmid DNA, however eae gene was not found in the strain. Although they produced VT, it seemed that the virulence of two strains were relatively weak because of the lack of the eae gene. In addition, the serotype O157 : H7 isolate resistant to ampicillin and streptomycin, while isolates of serotype O26 : H4 and O55 : Hl2 were multi-resistant to antibiotics including ampicillin, carbenicillin , cephalothin, trimethoprim/sulfamethoxazole and tetracycline. Supernatants of cultures of all three isolates were showed cytotoxic effect to vero and HeLa cell

• Korean Journal of Veterinary Research
• /
• v.33 no.4
• /
• pp.773-779
• /
• 1993

• Korean Journal of Veterinary Service
• /
• v.28 no.4
• /
• pp.407-412
• /
• 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.