本实验对大熊猫肠道分离的88株大肠杆菌、32株肺炎克雷伯氏菌对季铵盐类消毒剂BC、CTPC、CTAB及DDAC的最小抑菌浓度(MIC)值进行测定,并扩增了消毒剂的耐药基因。结果显示,大肠杆菌对季铵盐类消毒剂MIC值为:BC的MIC值介于8~128 mg/L;CTPC的MIC值在32~256 mg/L之间;CTAB的MIC值为64~512 mg/L;DDAC的MIC值介于8~128 mg/L。肺炎克雷伯氏菌对季铵盐类消毒剂的耐药情况为:BC的MIC值介于16~512 mg/L;CTPC的MIC值在64~256 mg/L之间;CTAB的MIC值介于128~512 mg/L;DDAC的MIC值介于8~64 mg/L。可见,肺炎克雷伯氏菌对季铵盐类消毒剂的MIC值要大于大肠杆菌对季铵盐类消毒剂的MIC值。耐药基因检测结果表明,大肠杆菌季铵盐类消毒剂的染色体型耐药基因扩增率为68.18%~98.86%,最高为sug E(98.86%),emr E最低(68.18%),没有检测出qac E、qac F、qac G,检出率最高的可移动遗传元件介导耐药基因为qac EΔ1(19.31%)。肺炎克雷伯氏菌的染色体型耐药基因检出率为13.64%~28.41%,ydg E最高(28.14%),emr E检出率最低(13.64%),可移动遗传元件介导耐药基因sug E(p)检出率最高(6.82%),qac EΔ1、qac F、qac G基因未检出。测定大熊猫源大肠杆菌及肺炎克雷伯氏菌对消毒剂的耐药性,对圈养大熊猫消毒剂的规范使用,防控大熊猫细菌性疾病以及细菌对消毒剂耐药性有着重要意义。
A total of 88 Escherichia coli and 32 Klebsiella pneumonia,isolated from 64 giant pandas from China Conservation Research Center for the Giant Panda Bifengxia base,were used to determine the minimum inhibitory concentrations( MICs) of quaternary ammonium compound( QAC) and disinfectants( BC,CTPC,CTAB and DDAC) using agar dilution method. The presence of QAC resistance genes were detected by PCR amplification with specific primers. The results showed that the MIC of QAC disinfectants in E. coli and K. pneumonia strains were different. The MICs of BC in E. coli were between 8 ~128 mg / L,CTPC were 32 ~ 256 mg / L,CTAB were 64 ~ 512 mg / L,and DDAC were 8 ~ 128 mg / L,respectively. The MICs of BC in K. pneumonia were 16 ~ 512 mg / L,CTPC were 64 ~ 256 mg / L,CTAB were 128 ~ 512 mg / L,and DDAC were 8 ~64 mg / L,respectively. Our results revealed that the MICs of K. pneumonia were higher than those of E. coli. The frequencies of chromosome-encoded resistant genes in E. coli were 68. 18% ~ 98. 86%,while emr E gene was the lowest and sug E gene was the highest. The most widespread mobile element encoded resistant gene was qac EΔ1. The frequencies of chromosome-encoded resistant genes in E. coli were 13. 64% ~ 28. 41%,while emr E gene was the lowest and ydg E gene was the highest. The most widespread mobile element encoded resistant gene was sug E( p). The detection of disinfectant resistance in E. coli and K. pneumonia may provide an important guidance for controlling the infection and bacterial resistance of giant pandas.