This study was conducted to investigate the presence of Escherichia (E.) coli O157 and E. coli O157:H7 and stx1 and stx2 genes on cattle carcasses and in rectal samples collected from Samsun Province of Turkey. A total of 200 samples collected from cattle carcasses and the rectal contents of 100 slaughtered cattle from two commercial abattoirs were tested using the immunomagnetic separation technique and multiplex PCR methods. E. coli O157 and E. coli O157:H7 were detected in 52 of the 200 samples (26%) tested. Of the positive samples, 49 were E. coli O157 and three were E. coli O157:H7. The E. coli O157 strain was isolated from 24 carcasses and 25 rectal samples, while E. coli O157:H7 was isolated from two carcasses and one rectal sample. Of the 49 samples positive for E. coli O157, 32 were from the rectal and carcass samples of the same animal, while two E. coli O157:H7 isolates were obtained from rectal swabs and carcasses of the same animal. The stx1 and stx2 genes were both detected in 35 E. coli O157 isolates and one E. coli O157:H7 isolate, but the stx2 gene was only detected alone in two E. coli O157 isolates. Overall, 16 carcasses tested positive for E. coli O157 and one carcass tested positive for E. coli O157:H7 based on both carcass and rectal samples. Overall, the results of this study indicate that cattle carcasses pose a potential risk to human health due to contamination by E. coli O157 and E. coli O157:H7 in the feces.
Abattoirs ; Animals ; Cattle ; Escherichia coli O157/*genetics/isolation & purification ; *Immunomagnetic Separation ; Meat/microbiology ; *Polymerase Chain Reaction ; Rectum/microbiology ; Shiga Toxin 1/*genetics ; Shiga Toxin 2/*genetics ; Turkey

China ; epidemiology ; Escherichia coli Infections ; epidemiology ; microbiology ; Escherichia coli O157 ; classification ; isolation & purification ; metabolism ; Humans ; Shiga Toxin 1 ; biosynthesis ; isolation & purification ; Shiga Toxin 2 ; biosynthesis ; isolation & purification ; Sorbitol

No Abstract Available.
Anti-Infective Agents* ; Enterohemorrhagic Escherichia coli* ; Quinolones* ; Shiga Toxins*

BACKGROUND: No outbreak of Enterohemorrhagic Escherichia coli (EHEC) infection has occurred as a group in Korea. On July 2004, an outbreak of EHEC infection occurred in an elementary school in Gwangju metropolitan city. Epidemic investigation was undertaken to track the source of infection and the mode of transmission of EHEC. METHODS: All students and staffs of the elementary school were interviewed and completed questionnaires. We surveyed their clinical symptoms and the foods that they ate. Microbiologic examinations were also carried out on the above school-related persons and many environmental specimens. We also investigated the facilities of the school, some suppliers of food materials, and other associated institutions. All the EHEC-positive persons were isolated in 5 hospitals and tested everyday for verotoxin until they turned out to be negative twice in succession, and their family were also interviewed and tested for EHEC. Pulsed-field gel electrophoresis (PFGE) was performed to find out the genetic relationship between isolates. RESULTS: Of the 1,643 school-related persons, 77 persons (4.7%) were positive for EHEC. Most of them were asymptomatic. All the isolated strains were non-O157 EHEC. Serotype O91 was the most frequent serotype (68 isolates), and the isolates revealing O91 serotypes showed identical PFGE patterns. The school meal was significantly associated with this outbreak (relative risk=13.29, p=0.00). CONCLUSIONS: This is the first EHEC outbreak occurred as a group in Korea, All the isolated strains were non-O157 serotypes and the mode of transmission was most likely by school meal.
Electrophoresis, Gel, Pulsed-Field ; Enterohemorrhagic Escherichia coli* ; Gwangju ; Humans ; Korea ; Meals ; Shiga Toxins ; Surveys and Questionnaires

A multiplex loop-mediated isothermal amplification (mLAMP) assay was developed for simultaneous detection of the stx1 and stx2 genes and applied for detection of shiga toxin-producing Escherichia coli (STEC) in cattle farm samples. Two target genes were distinguished based on T m values of 85.03 +/- 0.54degrees C for stx1 and 87.47 +/- 0.35degrees C for stx2. The mLAMP assay was specific (100% inclusivity and exclusivity), sensitive (with a detection limit as low as 10 fg/microL), and quantifiable (R 2 = 0.9313). The efficacy and sensitivity were measured to evaluate applicability of the mLAMP assay to cattle farm samples. A total of 12 (12/253; 4.7%) and 17 (17/253; 6.7%) STEC O157, and 11 (11/236; 4.7%) non-O157 STEC strains were isolated from cattle farm samples by conventional selective culture, immunomagnetic separation, and PCR-based culture methods, respectively. The coinciding multiplex PCR and mLAMP results for the types of shiga toxin revealed the value of the mLAMP assay in terms of accuracy and rapidity for characterizing shiga toxin genes. Furthermore, the high detection rate of specific genes from enrichment broth samples indicates the potential utility of this assay as a screening method for detecting STEC in cattle farm samples.
Animals ; Cattle ; Cattle Diseases/epidemiology/microbiology ; Escherichia coli Infections/epidemiology/microbiology/*veterinary ; Feces/microbiology ; Multiplex Polymerase Chain Reaction/veterinary ; Nucleic Acid Amplification Techniques/*veterinary ; Shiga Toxin 1/*genetics/isolation & purification ; Shiga Toxin 2/*genetics/isolation & purification ; Shiga-Toxigenic Escherichia coli/*genetics/isolation & purification

A total of 156 Shiga-like toxin producing Escherichia coli (STEC) were isolated from fecal samples of Korean native (100/568, 18%) and Holstein dairy cattle (56/524, 11%) in Korea between September 2010 and July 2011. Fifty-two STEC isolates (33%) harbored both of shiga toxin1 (stx1) and shiga toxin2 (stx2) genes encoding enterohemolysin (EhxA) and autoagglutinating adhesion (Saa) were detected by PCR in 83 (53%) and 65 (42%) isolates, respectively. By serotyping, six STEC from native cattle and four STEC from dairy cattle were identified as O-serotypes (O26, O111, O104, and O157) that can cause human disease. Multilocus sequence typing and pulsed-field gel electrophoresis patterns highlighted the genetic diversity of the STEC strains and difference between strains collected during different years. Antimicrobial susceptibility tests showed that the multidrug resistance rate increased from 12% in 2010 to 42% in 2011. Differences between isolates collected in 2010 and 2011 may have resulted from seasonal variations or large-scale slaughtering in Korea performed to control a foot and mouth disease outbreak that occurred in early 2011. However, continuous epidemiologic studies will be needed to understand mechanisms. More public health efforts are required to minimize STEC infection transmitted via dairy products and the prevalence of these bacteria in dairy cattle.
Animals ; Anti-Bacterial Agents/pharmacology ; Cattle/microbiology ; Drug Resistance, Multiple, Bacterial ; Electrophoresis, Gel, Pulsed-Field/veterinary ; Escherichia coli Infections/epidemiology/microbiology/*veterinary ; Female ; Genes, Bacterial/genetics ; Latex Fixation Tests/veterinary ; Microbial Sensitivity Tests/veterinary ; Multilocus Sequence Typing/veterinary ; Prevalence ; Republic of Korea/epidemiology ; Shiga Toxin 1/genetics ; Shiga Toxin 2/genetics ; *Shiga-Toxigenic Escherichia coli/drug effects/genetics

Verotoxin-producing Escherichia coli O157 is a primary cause of severe and bloody diarrhea. Campylobacter spp. are one of the commonly reported bacterial cause of gastrointestinal infections throughout the world. Only a few cases involving both E. coli O157 and Campylobacter species have been reported. The authors simultaneously isolated verotoxin-producing E. coli O157 and Campylobacter species from the stool of a 3 year-old male with bloody diarrhea, fever and abdominal pain.
Abdominal Pain ; Campylobacter* ; Child, Preschool ; Diarrhea ; Escherichia coli O157* ; Escherichia coli* ; Escherichia* ; Fever ; Gastroenteritis* ; Humans ; Male ; Shiga Toxins ; Shiga-Toxigenic Escherichia coli

BACKGROUND: Since 1982, many countries has reported outbreaks or sporadic cases caused by enterohaemorrhagic Escherichia coli (EHEC) serogroup strains, mainly E. coli O157:H7 type strain. However, systemic investigation about EHEC agents, including E. coli O157:H7, have not been done in Korea. Therefore, we investigated serogroup and verotoxin productivity of E. coli strains isolated from diarrheal patients and estimated risk of human infection in comparison with the EHEC strains isolated from cow, pig, and food material in Korea. METHODS: Diarrheal patient stool samples were collected and E. coli strains were isolated, according to biochemical characteristics. In order to isolate E. coli O157:H7, D-Sorbitol negative E. coli strains were selected. Serogrouping of the E. coli isolates was done by agglutination test. Verocytotoxin productivity was investigated by polymerase chain reaction (PCR) and reversed passive latex agglutination (RPLA). Human infection risk was estimated in comparison with EHEC strains isolated from cow, pig and food materials in Korea. RESULTS: Twenty-five E. coli strains were isolated from the diarrheal patients who were suspected to be infected with EHEC. However, none of these E. coli strains produced verocytotoxin. Out of 25 E. coli isolates, 16 serogroups of E. coli O1, O6, O8, O15, O20, O25, O26, O28, O29, O44, O86a, O119, O126, O128, O152 and 157:H- were found. In each of the E. coli O157:H- and O25 serogrorps 3 strains were found. CONCLUSION: None of 25 E. coli isolated from diarrheal patients who were suspected of EHEC infection produced verocytotoxin producing E. coli have been reported recently in Korea.
Agglutination ; Agglutination Tests ; Disease Outbreaks ; Efficiency* ; Enterohemorrhagic Escherichia coli ; Escherichia coli* ; Escherichia* ; Humans ; Korea* ; Latex ; Polymerase Chain Reaction ; Shiga Toxins

Escherichia (E.) coli serotype O157:H7 is a globally distributed human enteropathogen and is comprised of microorganisms with closely related genotypes. The main reservoir for this group is bovine bowels, and infection mainly occurs after ingestion of contaminated water and food. Virulence genetic markers of 28 O157:H7 strains were investigated and multilocus enzyme electrophoresis (MLEE) was used to evaluate the clonal structure. O157:H7 strains from several countries were isolated from food, human and bovine feces. According to MLEE, O157:H7 strains clustered into two main clonal groups designated A and B. Subcluster A1 included 82% of the O157:H7 strains exhibiting identical MLEE pattern. Most enterohemorrhagic E. coli (EHEC) O157:H7 strains from Brazil and Argentina were in the same MLEE subgroup. Bovine and food strains carried virulence genes associated with EHEC pathogenicity in humans.
Animals ; Argentina/epidemiology ; Brazil/epidemiology ; Cattle ; Cattle Diseases/epidemiology/*microbiology ; Enterohemorrhagic Escherichia coli/genetics/*isolation & purification/pathogenicity ; Escherichia coli O157/*genetics/*isolation & purification/pathogenicity ; Food Microbiology ; Gene Expression Regulation, Bacterial/physiology ; Genetic Markers ; Humans ; Polymerase Chain Reaction/veterinary ; Shiga Toxin 1/genetics/metabolism ; Shiga Toxin 2/genetics/metabolism ; Virulence

Enterohemorrhagic Escherichia coli (EHEC) is an important cause of bloody diarrhea in children, but is considered to be rare in infants. Herein, a case of infant hemorrhagic colitis of verotoxin-producing E. coli O157:H7 diagnosed by multiplex PCR is reported. A nine-month-old boy was admitted to our hospital with bloody diarrhea for the previous two days. Multiplex PCR using Seeplex(R) Diarrhea ACE Detection Kit (Seegene, Seoul, Korea) was directly applied to the stool specimens. Amplified bands specific for verotoxin, O157, and H7 indicated the presence of O157:H7 EHEC. The stool specimens were inoculated on sorbitol-MacConkey agar (SMA) and tryptic soy broth containing mitomycin C (TSB-M). Colorless colonies on sorbitol-MacConkey agar were O157-positive. TSB-M enrichment cultures of the stool specimen and the isolates were positive for verotoxin according to an enzyme immunoassay (EIA). The prepared ingredients of baby foods for the patient including ground meat, chopped carrot, chopped cabbage, and white rice porridge showed no EHEC on TSB-M and SMA. The patient's parents and three-year-old sister did not recently have any gastrointestinal symptoms. Cefdinir was administered for one day and was ceased after diagnosis of EHEC colitis. The stool culture and verotoxin assay were negative on the second day of hospitalization. Application of multiplex PCR and verotoxin EIA directly to diarrheal stool warrants the rapid diagnosis and appropriate treatment of EHEC colitis.
Agar ; Brassica ; Caseins ; Cephalosporins ; Child ; Colitis ; Daucus carota ; Diarrhea ; Enterohemorrhagic Escherichia coli ; European Continental Ancestry Group ; Hospitalization ; Humans ; Immunoenzyme Techniques ; Infant ; Meat ; Mitomycin ; Multiplex Polymerase Chain Reaction ; Parents ; Protein Hydrolysates ; Shiga Toxins ; Shiga-Toxigenic Escherichia coli ; Siblings