OBJECTIVETo type and group the Shiga-toxin producing Escherichia coli O157:H7 strains isolated recent years in China to understand the epidemiological features caused by the pathogen.

METHODSPulsed field gel electrophoresis of large restriction fragments of bacterial chromosomal DNA was used.

RESULTSThe 51 isolates of E. coli O157:H7 collected in recent years in China could be divided into 8 Pulsed Field Gel Electrophoresis (PFGE) types based on the size and number of restriction fragments and patterns, that were digested by XbaI. Strains isolated from Ningxia province showed only two types- PFGE1 and PFGE2. Strains isolated in Xuzhou of Jiangsu province had 6 PFGE types. Isolates identified between 1986 - 1988 belonged to PFGE7. Strains isolated from patients in 1999 - 2000 were PFGE5 and PFGE3. Strains isolated from stool samples of domestic animal, food and vegetable were PFGE3 - 6, of which the predominant type was PFGE5. All of the 5 strains isolated from patients with diarrhea and hemolytic uremic syndrome (HUS) belonged to PFGE type 5, which was the dominant type of the isolates from stool samples of domestic animal and samples of food and vegetable contaminated.

CONCLUSIONData suggested that the cluster patients with diarrhea and HUS might have been related to the pathogens from domestic animas and contaminated food or vegetables. The distribution of PFGE types also varied in different provinces of China.

Electrophoresis, Gel, Pulsed-Field ; Escherichia coli O157 ; classification ; genetics ; pathogenicity ; Genotype ; Humans ; Shiga Toxin ; biosynthesis

Enterohemorrhagic Escherichia coli serotype O157:H7 is a pathotype of diarrheagenic E. coli that produces one or more Shiga toxins, forms a characteristic histopathology described as attaching and effacing lesions, and possesses the large virulence plasmid pO157. The bacterium is recognized worldwide, especially in developed countries, as an emerging food-borne bacterial pathogen, which causes disease in humans and in some animals. Healthy cattle are the principal and natural reservoir of E. coli O157:H7, and most disease outbreaks are, therefore, due to consumption of fecally contaminated bovine foods or dairy products. In this review, we provide a general overview of E. coli O157:H7 infection, especially focusing on the bacterial characteristics rather than on the host responses during infection.
Animals ; Cattle ; Cattle Diseases/blood/epidemiology ; Developing Countries ; *Enterohemorrhagic Escherichia coli ; Escherichia coli Infections/blood/*epidemiology/veterinary ; *Escherichia coli O157/genetics/pathogenicity ; Feces/microbiology ; Hemolytic-Uremic Syndrome/blood/epidemiology/veterinary ; Operon ; Shiga Toxins/analysis ; Shigella dysenteriae ; Virulence

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) O157:H7 is an important pathogen. One of the important virulence traits of EHEC O157:H7 is the capacity to produce attaching and effacing (A/E) lesions on enterocyte. This property encoded by a pathogenicity island termed the locus of enterocyte effacement (LEE). LEE contains ler (LEE-encoded regulator) gene. The product of ler is a central up-regulator of many virulence genes of the LEE. Another important virulence factor of EHEC O157: H7 is Shiga toxin (Stx), encoded by a prophage integrated into the chromosome of O157:H7. In order to obtain an attenuated vaccine candidate, a ler deletion mutant of O157: H7 was constructed by use of suicide vector pCVD442. Meanwhile, due to potential instability of the prophage carrying the stx gene, the prophage was cured with serial passages of bacteria and confirmed by PCR and DNA sequencing. A ler/stx deletion mutant of EHEC O157:H7 was constructed, termed as O157:H7(deltaler/deltastx). The cultural supernatant of O157 ler/stx deletion mutant was inoculated in vero cell culture, and the result indicating that O157 ler/stx deletion mutant lost the toxigenicity to vero cell. Test group and control group of mice were orogstrically inoculated with the O157 ler/stx deletion mutant and the virulent strain O157:H7 EDL933, respectively. Mice were observed daily for clinical signs and weight changes. After inoculation of the deletion mutant, test group of mice (inoculated with O157:H7(deltaler/deltastx)) gained weight normally and experienced no clinical signs. In contrast, control group of mice (inoculated with O157: H7) exhibited weight loss and all died in four days. In another experiment, pregnant mice were orally vaccinated by O157:H7(deltaler/ deltastx) twice at interval of 14 days. Subsequently, the suckling mice were orally challenged with O157:H7 EDL933 at 7 days of age. The results showed that 78.34% of the sucking mice born by vaccinated mice were survival and 12.73% of the sucking mice born by non-vaccinated mice were survival. This study demonstrated that O157 ler/stx deletion mutant lost the toxigenicity to vero cell and to be safety to mice. Oral immunization can induce specific immune responses, and this mutant strain could be used as an attenuated vaccine candidate against EHEC O157.
Administration, Oral ; Animals ; Animals, Suckling ; Antibodies, Bacterial ; blood ; immunology ; Cercopithecus aethiops ; Escherichia coli Infections ; immunology ; mortality ; prevention & control ; Escherichia coli O157 ; genetics ; immunology ; pathogenicity ; Escherichia coli Proteins ; genetics ; Escherichia coli Vaccines ; administration & dosage ; genetics ; immunology ; Female ; Gene Deletion ; Genetic Vectors ; genetics ; Genomic Islands ; genetics ; Immunization ; Lactation ; immunology ; Male ; Mice ; Mutation ; Pregnancy ; Shiga Toxin ; genetics ; Survival Rate ; Trans-Activators ; genetics ; Vaccines, Attenuated ; administration & dosage ; genetics ; immunology ; Vero Cells ; Virulence ; genetics