Objective To analyze the subtypes of eae genes in various non-O157 Shiga toxin-pro-ducing Escherichia coli ( STEC) strains isolated in China.Methods The complete nucleotide sequences of 10 eae genes were amplified by PCR and sequenced.The BLASTn software was used to analyze the se-quences for eae gene subtyping.A phylogenetic tree was constructed based on the10 ea e gene sequences to-gether with the gene sequences of 30 different subtypes in GenBank and those of STEC strains of 7 prevalent serotypes (O157 ∶H7, O26 ∶H11, O103 ∶H2, O111 ∶H8, O145 ∶H28, O45 ∶H2 and O121 ∶H19) using MEGA 5.0.Multilocus sequence typing (MLST) was performed on the 10 STEC strains with reference to the Escherichia coli ( E.coli) MLST website ( http://mlst.warwick.ac.uk/mlst/dbs/Ecoli) for the typing of multiple loci.A minimum spanning tree ( MST) was constructed using the BioNumerics software to inves-tigate the phylogenetic relationships between the 10 eae gene-positive STEC strains in this study and hemolyt-ic uremic syndrome-associated enterohemorrhagic E.coli ( HUSEC) strains as well as all human STEC strains of O157, O26, O45, O103, O111, O121 and O145 serotypes submitted to the E.coli MLST website data-base.Results The complete nucleotide sequences of eae genes in 10 non-O157 STEC strains were 2.8 kb in length and belonged to 3 known subtypes.The predominant subtype wasβ1, accounting for 60%of the 10 STEC strains (6/10), followed byθandγ1 subtypes with two strains in each type.The eae gene sequences in certain strains were identical to those of the prevalent serotypes.Seven sequence types ( STs) were identi-fied from the 10 STEC strains carrying eae gene.Conclusion The eae genes harbored by the non-O157 STEC strains isolated from different specimens in China were diverse and had close phylogenetic relationships with the highly pathogenic and prevalent STEC strains.This study implied that the STEC strains harboring eae gene had high pathogenic potential.

Objective To investigate the tellurite resistance level,the presence of tellurite resistance (ter) gene cluster and their relationships in non-O157 Shiga toxin-producing Escherichia coli(STEC) isolates.Methods Tellurite resistance level was evaluated by plate dilution method and the ter gene cluster was tested by PCR.Results Only 5 of 39 non-O157 STEC isolates tested in this study were identified to have ter gene cluster,which showed relatively high levels of tellurite resistance ranging from 128 μg/ml to 512 μg/ml.In contrast,the other 34 isolates without ter gene cluster were sensitive to potassium tellurite and showed very low levels of tellurite resistance,the minimal inhibitory concentration (MIC) was ＜1 μg/ml for 29 isolates,8 μg/ml for 2 isolates and 2 μg/ml for 3 isolates.Conclusion Most non-O157 STEC isolates were sensitive to potassium tellurite.It could be concluded that much attention should be paid when screening the non-O157 STEC isolates using the selective medium supplemented with potassium tellurite.

Objective To understand the molecular characteristics of human-derived non-O157 Shiga toxin-producing Escherichia coil (STEC) strains circulating in five regions of China.Methods Twenty-seven non-O157 STEC strains isolated in five geographic regions were investigated by serotyping, stx1/stx2 subtyping and PCR screening for adhesion and other virulence genes.A multilocus sequence typing (MLST) scheme provided by E.coil MLST database were performed to amplify and sequence seven housekeeping genes (adk, icd, fumC, rgyrB, purA, mdh and recA) in those strains.Results Twenty-seven non-O157 STEC strains were typed into 16 O∶H serotypes.Among those strains, 11 harbored stx1a, 12 harbored stx1c, two harbored stx2e and the other three strains respectively harbored stx1a+stx2b, stx2d and stx2g.Positive rates of eae, efa1, saa, paa, toxB, astA and ehxA genes were 18.5%, 18.5%, 29.6%, 22.2%, 11.1%, 11.1% and 25.9%, respectively.The 27 strains were typed into 16 different sequence types (STs) based upon MLST.Conclusion Human-derived non-O157 STEC strains circulating in five regions of China are heterogeneous in their serotypes, stx1/stx2 subtypes and virulence gene profiles.

OBJECTIVETo investigate the molecular typing feature of enteropathogenic Escherichia coli (EPEC) strains isolated from different reservoirs in eight provinces of China from 2006 to 2014.

METHODSAccording to the time, place, reservoir, and PFGE pattern of the EPEC strains isolated from stools of humans with diarrhea, animal feces, and foods in eight provinces of China between 2006 and 2014, 149 EPEC strains were selected and characterized by multilocus sequence typing (MLST) using seven housekeeping genes provided by E.coli MLST database. Strain analysis demonstrated 56 different sequence types (STs). SeqMan II, MEGA 5.05, and eBURST V3 were applied to analyze the genetic relationships of domestic and forein existing 392 strains (243 EPEC strains included in the E.coli MLST database and 149 EPEC strains comprised in the present study).

RESULTSAmong the 56 different STs, the prevalent ST was ST-40, which included 19 (19/149, 12.8%) isolates. Nineteen new STs were identified. Eleven new alleles were detected in six house-keeping genes (adk, fumC, gyrB, icd, mdh, and purA). Six STs were simultaneously detected among EPEC strains isolated from patients with diarrhea and animals. And these EPEC strains were all aEPEC strains. Two STs were simultaneously identified among EPEC strains isolated from patients with diarrhea and foods. Also, these EPEC strains were all aEPEC strains. 33 out of 173 STs were divided into five major clone complexes by eBURST, STC-29, STC-10, STC-20, STC-28, and STC-517. The remaining EPEC strains included in the other 140 STs were part of the other clone complexes or just were singletons.

CONCLUSIONA high degree of phylogenetic heterogeneity was observed among the EPEC strains isolated in eight provinces of China. The EPEC strains with same STs of human isolates isolated from animal feces and foods were all aEPEC strains.

Animals ; China ; Diarrhea ; Enteropathogenic Escherichia coli ; Escherichia coli ; Escherichia coli Proteins ; Feces ; Humans ; Multilocus Sequence Typing ; Phylogeny

Shiga toxin (Stx), which can be divided into Shiga toxin 1 (Stx1) and Shiga toxin 2 (Stx2), is an important virulence factor of Shigella spp. and certain strains of Escherichia coli. Stx, enco-ded by λ-like phage, blocks protein synthesis through removal of an adenine residue from the 28S rRNA. Stx can also induce apoptosis through multiple pathways. Humans may suffer from diarrhea, hemorrhagic colitis ( HC) and hemolytic uremic syndrome ( HUS) and even death when infected with Shiga toxin-producing bac-teria. At present, there is no specific treatment for diseases caused by Stx. In recent years, the application of Stx in cancer therapy and imaging has aroused great interest. This review provided a brief overview of Stx in its nomenclature, typing, structure, genetics, pathogenesis and application perspectives.

Escherichia albertii is an emerging zoonotic enteropathogen which mainly causes infectious diarrhea. Since the discovery and naming of Escherichia albertii, it was found to be responsible for several outbreaks of foodborne gastroenteritis and widely distributed in avian and wild animals. Due to the lack of specific identification system, the global Escherichia albertii infections might be underestimated. Though avian has been considered as the important reservoir of Escherichia albertii, its role in disease transmission remains unclear. This study reviewed the biochemical properties, genomic characteristics, isolation and identification methods of Escherichia albertii, and its prevalence in human, host animals and food. The risk of Escherichia albertii infection and future perspectives in this field were also discussed.

Objective:To understand the molecular characteristics of Escherichia coli producing Shiga toxin 2e subtype isolated from different sources in China. Methods:Three human-derived, 13 animal-derived and eight food-derived stx2e-positive Escherichia coli strains which were isolated during 2012 to 2018 were analyzed by antimicrobial susceptibility testing and whole genome sequencing. The stx subtype, serotype, multi-locus sequence type, virulence genes and antimicrobial resistance genes of each strain were determined by whole genome sequences. The phylogenetic relationship and genetic composition of Shiga-toxin prophage were explored. Results:Twenty-four stx2e-STEC strains were typed into 19 O∶H serotypes and 19 sequence types (STs). Each strain carried at least one kind of antimicrobial resistance gene and 19 out of 24 strains were resistant to at least one kind of antimicrobials. Three human-derived strains were heterogenous in serotypes and STs, but there were several animal and food-derived strains shared the same serotype or ST with human strains and showed close relationship in the phylogenetic analysis. The sequences of stx2e among all strains were highly conserved (similarity >99.7%), but there were significant differences in the size and the gene composition of Shiga toxin prophage genome. Conclusions:This is report about the characteristics of rare human-derived Stx2e-STEC strains in China. Comparing human isolates with animal-and food-derived strains, it indicates that Stx2e-STEC strains are highly genetic diversity and have the potential to infect humans.