Masco L, Van Hoorde K, De Brandt E, Swings J, Huys G
Laboratory of Microbiology, Department of Biochemistry, Physiology and Microbiology, Ghent University K.L. Ledeganckstraat 35, B-9000 Gent, Belgium.
J Antimicrob Chemother. 2006 Jul;58(1):85-94. doi: 10.1093/jac/dkl197. Epub 2006 May 12.
The aim of this study was to assess the antimicrobial susceptibility of a taxonomically diverse set of Bifidobacterium strains to different classes of antimicrobial agents using a recently described medium.
The susceptibility of 100 strains encompassing 11 bifidobacterial species originating from humans, animals and probiotic products to 12 antimicrobial agents was tested by agar overlay disc diffusion. Based on these results, one or two strains per species were selected for susceptibility testing to nine antibiotics by broth microdilution using the Lactic acid bacteria Susceptibility test Medium (LSM) supplemented with cysteine. The genotypic basis of atypical tetracycline resistance was further characterized using PCR, Southern blotting and partial sequencing.
Based on the distribution of inhibition zone diameters and MIC values, all strains tested were susceptible to amoxicillin, chloramphenicol, erythromycin, quinupristin/dalfopristin, rifampicin and vancomycin. Our data also reinforce earlier observations indicating that bifidobacteria are intrinsically resistant to gentamicin, sulfamethoxazole and polymyxin B. Susceptibility to trimethoprim, trimethoprim/sulfamethoxazole, ciprofloxacin, clindamycin, tetracycline and minocycline was variable. The tet(W) gene was responsible for tetracycline resistance in 15 strains including 7 probiotic isolates belonging to the taxa Bifidobacterium animalis subsp. lactis and Bifidobacterium bifidum. This gene was present in a single copy on the chromosome and did not appear to be associated with the conjugative transposon TnB1230 previously found in tet(W)-containing Butyrivibrio fibrisolvens.
The use of the LSM + cysteine medium allowed us to discriminate between intrinsic and atypical resistance properties of bifidobacteria and sets the scene for future definition of epidemiological cut-off values for all important Bifidobacterium species. The presence of an acquired tet(W) gene in several probiotic product isolates stresses the need for a minimal safety evaluation during the selection of Bifidobacterium strains for probiotic use.
本研究旨在使用一种最近描述的培养基,评估一组分类学上多样的双歧杆菌菌株对不同类别的抗菌剂的药敏性。
通过琼脂覆盖纸片扩散法,测试了源自人类、动物和益生菌产品的11种双歧杆菌的100株菌株对12种抗菌剂的药敏性。基于这些结果,每个物种选择一到两株菌株,使用添加了半胱氨酸的乳酸菌药敏试验培养基(LSM),通过肉汤微量稀释法测试对九种抗生素的药敏性。使用PCR、Southern印迹和部分测序进一步表征非典型四环素抗性的基因型基础。
根据抑菌圈直径和MIC值的分布,所有测试菌株对阿莫西林、氯霉素、红霉素、奎奴普丁/达福普汀、利福平和万古霉素敏感。我们的数据还强化了早期的观察结果,表明双歧杆菌对庆大霉素、磺胺甲恶唑和多粘菌素B具有内在抗性。对甲氧苄啶、甲氧苄啶/磺胺甲恶唑、环丙沙星、克林霉素、四环素和米诺环素的敏感性各不相同。tet(W)基因导致15株菌株对四环素耐药,其中包括7株属于动物双歧杆菌乳酸亚种和两歧双歧杆菌的益生菌分离株。该基因在染色体上以单拷贝形式存在,似乎与先前在含tet(W)的溶纤维丁酸弧菌中发现的接合转座子TnB1230无关。
使用LSM + 半胱氨酸培养基使我们能够区分双歧杆菌的内在抗性和非典型抗性特性,并为未来确定所有重要双歧杆菌物种的流行病学临界值奠定了基础。几种益生菌产品分离株中存在获得性tet(W)基因,强调了在选择用于益生菌用途的双歧杆菌菌株时进行最低限度安全性评估的必要性。