›› 2012, Vol. 39 ›› Issue (5): 187-191.

• 疾病防治 • 上一篇    下一篇

3种四环素耐药基因型肉牛肠道大肠杆菌对四环素类抗生素的耐药性分析

金鑫, 苏蕊, 张文广, 张燕军, 王瑞军, 李金泉   

  1. 内蒙古农业大学动物科技学院, 内蒙古呼和浩特 010018
  • 收稿日期:1900-01-01 修回日期:2012-03-05 出版日期:2012-05-20 发布日期:2012-05-20
  • 通讯作者: 李金泉

Analysis on Antimicrobial Susceptibility to Tetracycline Analogues for Three Genotypes of Tetracycline Resistance in Escherichia coli Isolates from Feedlot Cattle

JIN Xin, SU Rui, ZHANG Wen-guang, ZHANG Yan-jun, WANG Rui-jun, LI Jin-quan   

  1. College of Animal Science, Inner Mongolia Agricultural University, Huhhot 010018, China
  • Received:1900-01-01 Revised:2012-03-05 Online:2012-05-20 Published:2012-05-20

摘要: 以低于治疗水平的氯四环素(CT)及低于治疗水平的氯四环素和治疗水平的氧四环素组合(CT-OX)两种方式分别对肉牛进行抗生素处理,研究其对肠道大肠杆菌耐药基因型的影响。从粪便样品分离大肠杆菌,并通过抗菌药物纸片法和稀释法敏感性试验测试分离出的大肠杆菌对四环素、氧四环素和氯四环素的敏感性。利用针对耐药基因tet(A)tet(B)tet(C)的引物对176个四环素耐药或中介的细菌样品进行多重PCR试验,结果发现所有样品均携带一种或两种耐药基因,tet(A)在两组样品中的流行基本相同,但CT组中tet(B)的流行比例显著小于CT-OX组(P<0.05),而tet(C)的流行比例则显著CT-OX组(P<0.05)。同时,在对四环素表现为中介的52个样品检测结果中,发现其中92.3%携带tet(C)基因。另外,最小抑菌浓度值(MICs)结果表明,药物敏感性同时取决于四环素类别和耐药基因型两方面。利用real-time PCR在转录水平上对tet(C)基因进行分析,发现耐药型与中介型并非上游调控造成。对tet(C)基因的测序分析结果发现,耐药型的第1063位碱基由T突变为G。由上述数据可知,对肉牛的四环素饲喂种类可以影响到大肠杆菌的耐药基因流行。

关键词: 大肠杆菌; 耐药性; 四环素; 肉牛

Abstract: The effect of administering feedlot cattle subtherapeutic levels of chlortetracycline (CT) or CT and therapeutic levels of oxytetracycline (CT-OX) on resistance genotype in Escherichia coli was investigated. Isolated E. coli from fecal samples. Isolates were tested for susceptibility to tetracycline, oxytetracycline, and chlortetracycline using disk diffusion or broth microdilution. Detection of tet(A), tet(B) and tet(C) genes encoded by tetracycline-resistant isolates (n=176) were performed by multiplex PCR. All isolates encoded one or a combination of two resistance genes. Prevalence of tet(A) was similar between groups of E. coli, however prevalence of tet(B) was lower(P<0.05) and tet(C) was greater(P<0.05) in CT isolates. The nature of the tet determinants was further assessed in a group of intermediately tetracycline-resistant isolates (n=52). Minimum inhibitory concentrations showed that susceptibility was dependent on tetracycline analogue and the type of resistance determinant. The tet(C) gene was present in 92% of these isolates. Copies of tet(C) transcripts, analyzed by real-time PCR, indicated that up-regulation did not occur in tetracycline-resistant isolates when compared to intermediately-resistant isolates. However, sequence analysis of the tet(C) gene revealed a T→G substitution at position 1063 in resistant isolates that may have affected phenotype. These data provide insight into the relationship between the type of tetracycline analogue administered to cattle and the prevalence of resistance genes in E. coli.

Key words: Escherichia coli; antimicrobial resistance; tetracycline; feedlot cattle

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