Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University, Chengdu, Sichuan, 610065, PR China; Nanchong Professional Technic College, No. 94, Hongfa Road, Xiaolong Town, Gaoping District, Nanchong City, 637131, Sichuan Province, PR China.
Key Laboratory of Bio-Resource and Eco-Environment of Ministry of Education, Animal Disease Prevention and Food Safety Key Laboratory of Sichuan Province, College of Life Sciences, Sichuan University, Chengdu, Sichuan, 610065, PR China; College of Life Science and Engineering, Northwest Minzu University, Lanzhou, 73030, Gansu, PR China.
Microb Pathog. 2018 Oct;123:68-73. doi: 10.1016/j.micpath.2018.06.006. Epub 2018 Jun 19.
The objective of this study was to investigate the co-occurrence of biofilms and quinolone resistance in Salmonella enterica serotype Typhimurium mediated by IncHI2-type oqxAB-positive plasmids. Among the 40 Salmonella strains, we found that 27 isolates formed biofilms and displayed identical multidrug-resistance profiles to ciprofloxacin, doxycycline, sulfamethoxazole-trimethoprim, ampicillin and streptomycin, based on biofilm formation assays and antimicrobial susceptibility testing. In particular, a single S. Typhimurium isolate named SC523 produced the thickest biofilms and exhibited the highest-level resistance (MIC = 8 μg/mL) to ciprofloxacin compared to those of the other isolates. The detection of known plasmid-mediated quinolone resistance (PMQR) genes and point mutations in the quinolone resistance-determining region (QRDR) by PCR assay showed that oqxAB genes were present in 27 biofilm-positive isolates. Conjugation experiments, S1-pulse-field gel electrophoresis and biofilm formation assays demonstrated that the conjugative plasmid that encoded biofilms and quinolone resistance in Salmonella SC523 could be transferred to a recipient with a frequency of 4.7 × 10 per recipient cell. The results of PCR-based replicon typing (PBRT) showed that the IncHI2-type plasmids accounted for 100% of the biofilm-oqxAB-positive isolates and transconjugants. The sequence analysis of Salmonella SC523 confirmed that the oqxAB cassette and fourteen DNA transfer genes in the IncHI2-type oqxAB-positive conjugative plasmid were genetically responsible for the phenotypic quinolone resistance and biofilm formation. The conclusion is that the IncHI2-type plasmid in S. Typhimurium isolate from chicken farm was identified and sequenced, which contained oqxAB and tra/trh and encoded quinolone resistance and biofilms, and could be transferred to recipients through conjugation. Notably, the prevalence of IncHI2-type biofilm-oqxAB-positive plasmids in animal-origin Salmonella poses a threat to public health, as these Salmonella from poultry farms show a decreased susceptibility to quinolones and could spread to humans.
本研究旨在调查由 IncHI2 型 oqxAB 阳性质粒介导的肠沙门氏菌血清型 Typhimurium 中生物膜与喹诺酮耐药的共同发生。在 40 株沙门氏菌菌株中,我们发现 27 株分离株通过生物膜形成试验和抗菌药物敏感性试验,形成生物膜并表现出对环丙沙星、强力霉素、磺胺甲恶唑-甲氧苄啶、氨苄西林和链霉素相同的多药耐药谱。特别是,一株名为 SC523 的单一致鼠伤寒沙门氏菌产生的生物膜最厚,与其他分离株相比,对环丙沙星的耐药水平最高(MIC=8μg/ml)。PCR 检测已知的质粒介导的喹诺酮耐药(PMQR)基因和喹诺酮耐药决定区(QRDR)的点突变表明,27 株生物膜阳性分离株均存在 oqxAB 基因。接合实验、S1 脉冲场凝胶电泳和生物膜形成试验表明,编码鼠伤寒沙门氏菌生物膜和喹诺酮耐药的可接合质粒可通过 4.7×10 个受体细胞的频率转移到受体中。基于 PCR 的复制子分型(PBRT)的结果表明,IncHI2 型质粒占生物膜-oqxAB 阳性分离株和转导子的 100%。鼠伤寒沙门氏菌 SC523 的序列分析证实,IncHI2 型 oqxAB 阳性可接合质粒中的 oqxAB 盒和 14 个 DNA 转移基因在遗传上负责表型喹诺酮耐药和生物膜形成。结论是,从养鸡场分离到的鼠伤寒沙门氏菌的 IncHI2 型质粒被鉴定和测序,该质粒包含 oqxAB 和 tra/trh,并编码喹诺酮耐药和生物膜,可通过接合传递给受体。值得注意的是,动物源性沙门氏菌中 IncHI2 型生物膜-oqxAB 阳性质粒的流行对公共卫生构成威胁,因为这些来自家禽养殖场的沙门氏菌对喹诺酮类药物的敏感性降低,并且可能传播给人类。