216 avian pathogenic Escherichia coli (APEC) isolates were obtained from poultry with colibacillosis in different areas of China. Among them, 195 were serotyped as O78, O88, and O93. Thirteen virulence-associated genes, including fimC, iucD, iss, tsh, fyuA, irp2, eaeA, hlyE, colV, papC, stx2f, vat, and astA, were submitted to PCR amplification. The fimC gene was the most prevalent with a detection rate of 93.6%, followed by iucD (70.8%), iss (58.8%), and tsh (51.4%) in APEC isolates. The detection rate of high pathogenicity islands (HPI)-associated fyuA and irp2 genes were both 44.9%, with no LEE (the locus of enterocyte effacement) island-associated gene eaeA detected. In terms of distribution patterns of the 13 virulence-associated genes, 5 isolates harborbed 10 genes, 19 isolates contained only fimC gene, and only 4 isolates had no virulence-associated gene detected. Different correlations of the virulence-associated genes with O serotypes were also investigated and 50% O78 isolates had a gene distribution patterns of fimC+iucD+irp2+fyuA+iss+colV+tsh+.
JIN Wen-jie ZHENG Zhi-ming QIN Ai-jian SHAO Hong-xia LIU Yue-long WANG Jiao WANG Qian-qian
Fosfomycin, a broad-spectrum antibiotic against both Gram-positive and Gram-negative bacteria, is very important in the clinic but many fosfomycin-resistant bacteria have been isolated from patients. In this study, the resistance mechanism of three fosfomycin-resistant avian pathogenic Escherichia coli (APEC) strains (JE1, IF7 and CD11) isolated from septicemic chickens were analyzed. The results showed that their fosfomycin-resistance mechanisms were different. An alteration in the glpT transport system was the main reason of the fosfomycin-resistance mechanisms of strain IF7. Compared with the control stain BL21, the capacity of fosfomycin-uptake was low in all these three stains (JE1>IF7>CD11). Sequence results of murA showed that there were more than 10 sites of nucleotide mutation, but only one amino acid mutation T116A showed in CD11. Real-time detection test showed that the expression level of the murA gene of the three stains was significantly increased (four times increase in strain CD11 and two times increase in strains JE1 and IF7). The transformation and recombinant test showed that the recombinant bacteria with the murA of JE1 and CD11 showed high minimal inhibitory concentration (MIC) against fosfomycin. From the results of this research, it showed that most of the fosfomycin-resistance mechanisms once showed in patient bacteria have appeared in the APEC strains and the fosfomycin-resistance mechanism of the three APEC isolates was different.
JIN Wen-jieZHENG Zhi-mingWANG Qian-qianQIN Ai-jianSHAO Hong-xiaQIAN Kun
