北京地区宠物犬源blaCTX-M基因阳性肺炎克雷伯菌的耐药表型和分子特征研究

doi:10.16431/j.cnki.1671-7236.2019.11.034

摘要/Abstract

摘要： 为研究宠物犬源bla_CTX-M基因阳性肺炎克雷伯菌的携带情况，同时了解宠物犬携带bla_CTX-M基因肺炎克雷伯菌的耐药表型和分子特征，本研究从355只宠物犬的鼻拭子和肛拭子中分离头孢噻肟耐药细菌，通过16S rDNA和基质辅助激光解吸电离飞行时间质谱鉴定细菌种属。对鉴定为肺炎克雷伯菌的菌株，运用PCR检测bla_CTX-M基因，药物敏感性试验测定bla_CTX-M基因阳性肺炎克雷伯菌的耐药表型，PFGE分型和全基因组测序技术分析bla_CTX-M基因阳性肺炎克雷伯菌的分子特征。PCR结果显示，共分离到7株bla_CTX-M基因阳性肺炎克雷伯菌，3株来自宠物肛拭子，4株来自宠物鼻拭子；药敏试验结果显示，7株菌均对头孢噻呋、头孢噻肟和阿莫西林-克拉维酸耐药，但对多黏菌素E、美罗培南和替加环素敏感。PFGE结果显示，其中3株细菌相似度为100%，而另外4株细菌相似度差异较大（<30%）。全基因组结果显示，7株菌共有5个ST分型，bla_CTX-M基因分离的亚型为CTX-M-14型（n=5）和CTX-M-3型（n=2），同时这些菌株中还携带了其他β-内酰胺类、氨基糖苷类和氟喹诺酮类耐药基因。从本研究结果可以看出，目前宠物犬源bla_CTX-M基因阳性肺炎克雷伯菌的检出率较低，但其多重耐药性较严重，本研究分离的耐药基因bla_CTX-M、bla_SHV亚型与国内外流行有差异。目前宠物在抗生素的使用和监控上仍有不足，需进一步关注宠物细菌耐药性，为后续防控提供依据。

关键词: 宠物犬; 肺炎克雷伯菌; CTX-M

Abstract: In order to investigate the presence,resistant phenotype and molecular characterization of bla_CTX-M gene positive Klebsiella pneumoniae in pet dogs,we isolated ceftaxime-resistant bacteria from anal and nasal swabs of 355 pet dogs and species were further identified by 16S rDNA and MALDI-TOF-MS.We detected bla_CTX-M gene by PCR.All bla_CTX-M gene carrying Klebsiella pneumoniae were subjected to antimicrobial susceptibility testing to observe resistance phenotype,PFGE typing and the whole sequencing to know molecular characterization.The results of PCR showed that 7 bla_CTX-M gene containing Klebsiella pneumoniae were isolated,of which three were from anal swabs and four were from nasal swabs.All strains were resistant to ceftiofur,cefotaxime and amoxicillin-clavulanate,but were susceptible to colistin,meropenem and tigecycline.The results of PFGE patterns indicated that the similarity of three isolates were 100% and the similarity of the other four were <30%.7 isolates belonged to 5 ST types,and 5 were bla_CTX-M-14 gene positive and 2 were bla_CTX-M-3 gene positive.However,some β-lactamase-resistant genes,quinolones-resistant genes and aminoglycosides-resistant genes were co-exist with bla_CTX-M gene in these strains.In summary,we found that the presence of bla_CTX-M gene carrying Klebsiella pneumoniae had a low detection rate,but they were multi-drug resistant,the phenotype of bla_CTX-M and bla_SHV genes in this study were different from the prevalence phenotype at home and abroad.Until now,the usage and control of antibiotics in companion animals still had limitation,we needed pay more attention to antibiotic resistance in companion animals to provide scientific evidence to control in the future.

Key words: pet dogs; Klebsiella pneumoniae; CTX-M

中图分类号:

S858.292

邹之宇, 雷蕾, 何俊佳, 陈绮妍, 邵兵, 汪洋. 北京地区宠物犬源bla_CTX-M基因阳性肺炎克雷伯菌的耐药表型和分子特征研究[J]. 中国畜牧兽医, 2019, 46(11): 3432-3439.

ZOU Zhiyu, LEI Lei, HE Junjia, CHEN Qiyan, SHAO Bing, WANG Yang. Resistant Phenotype and Molecular Characterization of bla_CTX-M Gene Carrying Klebsiella pneumoniae of Pet Dogs in Beijing[J]. China Animal Husbandry and Veterinary Medicine, 2019, 46(11): 3432-3439.

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