食品科学 ›› 2020, Vol. 41 ›› Issue (18): 77-84.doi: 10.7506/spkx1002-6630-20190802-026

• 生物工程 • 上一篇    下一篇

IncI1和IncN质粒阳性沙门氏菌耐药及质粒接合转移特征

盛焕精,李怡谰,王泽维,牛沁雅,孟令缘,曹晨阳,李伟,廉鲁昕,杨保伟   

  1. (西北农林科技大学食品科学与工程学院,陕西?杨凌 712100)
  • 出版日期:2020-09-25 发布日期:2020-09-18
  • 基金资助:
    国家自然科学基金面上项目(31671956)

Characteristics of Antibiotic Resistance and Plasmid Conjugative Transfer of IncI1 and IncN Plasmid Positive Salmonella

SHENG Huanjing, LI Yilan, WANG Zewei, NIU Qinya, MENG Lingyuan, CAO Chenyang, LI Wei, LIAN Luxin, YANG Baowei   

  1. (College of Food Science and Engineering, Northwest A&F University, Yangling 712100, China)
  • Online:2020-09-25 Published:2020-09-18

摘要: 目的:研究分离于北京、上海、福建、河南、四川、广东、广西、陕西和新疆等地各类零售食品、临床病人和食品性动物源沙门氏菌中IncI1和IncN质粒的流行状况,IncI1和IncN质粒阳性沙门氏菌的药敏性、携带的耐药基因及其在接合过程的水平转移状况。方法:使用聚合酶链式反应(polymerase chain reaction,PCR)复制子分型方法筛选携带不相容质粒IncI1和IncN的菌株,琼脂稀释法测定沙门氏菌的药敏性,PCR法检测耐药基因,膜接合法测定IncI1和IncN质粒的水平转移。结果:956?株沙门氏菌中共检出IncI1阳性菌株42?株(4.39%),IncN阳性菌株3?株(0.31%);26?株IncI1和/或IncN质粒阳性代表菌株对头孢噻呋(100.0%)、萘啶酮酸(92.3%)、氨苄西林(92.3%)、头孢哌酮(88.5%)、四环素(84.6%)、头孢曲松(80.8%)、复方新诺明(76.9%)、链霉素(76.9%)和氯霉素(61.5%)耐药最为普遍,对卡那霉素(26.9%)、庆大霉素(23.1%)、多黏菌素B(23.1%)、环丙沙星(19.2%)、阿莫西林/克拉维酸(15.4%)、头孢西丁(11.5%)和阿米卡星(3.8%)的耐药性相对较低。IncI1质粒阳性菌株比IncN质粒阳性菌株的耐药谱更宽,对头孢类抗生素的耐受水平更高;IncN质粒阳性菌株对氨基糖苷类抗生素的抗性相对更高。IncI1阳性菌株中blaTEM和blaCTX-M基因检出率高于IncN阳性株,IncN阳性株中qnrA、qnrB和qnrS基因检出率高于IncI1阳性菌株。沙门氏菌作为受体时接合频率在3.2×10-5~2.0×10-3之间,大肠杆菌作为受体时接合频率在8.7×10-7~9.6×10-5之间。供体菌携带的qnrB、acc(6’)-Ib、blaTEM和blaCTX-M基因均可通过接合转移至受体菌,接合后受体菌获得卡那霉素、头孢噻呋、头孢曲松、氨苄西林、链霉素和庆大霉素抗性。结论:IncI1和IncN质粒阳性沙门氏菌检出率较低,菌株携带的质粒类型和相应的耐药表型存在一定关联,质粒携带的耐药基因可通过接合作用在不同种属细菌间传播,使受体菌获得耐药性。

关键词: 沙门氏菌;IncN质粒;IncI1质粒;耐药基因;接合转移

Abstract: Objective: In this study, the prevalence, antimicrobial susceptibility, antibiotic resistance genes and plasmid horizontal transfer of IncI1 and IncN plasmid positive Salmonella isolated from retail foods, clinical patients and food animals in the provincial-level regions of Beijing, Shanghai, Henan, Sichuan, Guangdong, Guangxi, Shaanxi and Xinjiang were investigated. Methods: The incompatible plasmids IncI1 and IncN in Salmonella were determined by a polymerase chain reaction (PCR)-based replicon typing method, the antimicrobial susceptibility was tested by the agar dilution method, and the antibiotic resistance genes were detected using PCR. The horizontal transfer of IncI1 and IncN plasmids was conducted via conjugation. Results: Forty-two IncI1 positive isolates (4.39%) and 3 IncN positive isolates (0.31%) were identified among the 956 strains of Salmonella. Of these, 26 representatives of IncI1 and/or IncN plasmid positive Salmonella showed the highest prevalence of antibiotic resistance to ceftiofur (100.0%), nalidixic acid (92.3%), ampicillin (92.3%), cefoperazone (88.5%), tetracycline (84.6%), ceftriaxone (80.8%), and sulfamethoxazole/trimethoprim (76.9%), streptomycin (76.9%) and chloramphenicol (61.5%) followed by kanamycin (26.9%), gentamicin (23.1%), polymyxin B (23.1%), ciprofloxacin (19.2%), amoxicillin-clavulanic acid (15.4%), cefoxitin (11.5%) and amikacin (3.8%). The IncI1 plasmid positive Salmonella exhibited broader antibiotic-resistance spectra and higher resistance to cephalosporins than the IncN plasmid positive one, whereas the IncN plasmid positive isolates were more resistant to aminoglycoside antibiotics. blaTEM and blaCTX-M were more frequently detected from the IncI1 positive Salmonella, while qnrA, qnrB and qnrS were more prevalent in the IncN positive isolates. The IncI1 and/or IncN plasmid in the donor strains could be transferred to the recipients Salmonella and Escherichia coli at conjugation frequencies ranging from 3.2 × 10-5 to 2.0 × 10-3 and 8.7 × 10-7 to 9.6 × 10-5 transconjugant per recipient cell, respectively. The antibiotic resistance genes qnrB, acc(6’)-Ib, blaTEM and blaCTX-M could be transferred from the donors to the recipients during conjugation to impart the recipients with antibiotic resistance phenotypes to kanamycin, ceftiofur, ceftriaxone, ampicillin, streptomycin, and gentamicin after conjugation. Conclusion: The prevalence of IncI1 and IncN plasmid positive Salmonella was relatively low, and the antibiotic resistance phenotypes of the hosts were found to be associated with the types of plasmids they carried to some extent. The antibiotic resistance genes in the plasmids could be transferred from the donor to the recipient by horizontal gene flow among different species to confer antibiotic resistance to the recipient.

Key words: Salmonella; IncN plasmid; IncI1 plasmid; antibiotic resistance genes; conjugative transfer

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