Marine Emanuele, Pos Klaas M
Institute of Biochemistry, Goethe University Frankfurt, Frankfurt am Main, Germany.
Bio Protoc. 2020 Dec 20;10(24):e3865. doi: 10.21769/BioProtoc.3865.
, an obligate predatory bacterium [, bacteria that kill and feed on other bacteria (prey)], has the potential to be used as a probiotic for the disinfection of surfaces or for the treatment of bacterial infections. One option is to use this organism in combination with antimicrobials to potentiate the effectiveness of treatments. In order to make this approach feasible more has to be known about the ability of to resist antibiotics itself. Standard assays to determine the minimum inhibitory concentration (MIC) are not suitable for , since the small size of this bacterium (0.25-0.35 by 0.5-2 μm) prevents scattering at OD. Since these predatory bacteria require larger prey bacteria for growth ( dimensions are 1 by 1-2 μm), the basis for the antimicrobial sensitivity assay described here is the reduction of the OD caused by prey lysis during growth. Previous studies on predatory bacteria resistance to antimicrobials employed methods that did not allow a direct comparison of antimicrobial resistance levels to those of other bacterial species. Here, we describe a procedure to determine sensitivity to antimicrobials which can be compared to a reference organism tested as close as possible to the same experimental conditions. Briefly, minimal inhibitory concentration (MIC) values of are determined by measuring the reduction in absorbance at 600 nm of mixed predator/prey cultures in presence and absence of different antimicrobial concentrations. Of note, this method can be modified to obtain antimicrobial MIC values of other predatory bacteria, using different conditions, prey bacteria and/or antimicrobials.
[具体细菌名称]是一种专性捕食性细菌[即杀死并以其他细菌(猎物)为食的细菌],有潜力用作表面消毒或治疗细菌感染的益生菌。一种选择是将这种微生物与抗菌剂联合使用,以增强治疗效果。为了使这种方法可行,必须更多地了解[具体细菌名称]自身对抗生素的耐药能力。用于确定最低抑菌浓度(MIC)的标准检测方法不适用于[具体细菌名称],因为这种细菌体积小(0.25 - 0.35×0.5 - 2μm),无法在OD处产生散射。由于这些捕食性细菌生长需要更大的猎物细菌(尺寸为1×1 - 2μm),这里描述的抗菌敏感性检测的基础是生长过程中猎物裂解导致的OD降低。以往关于捕食性细菌对抗菌剂耐药性的研究采用的方法无法将抗菌耐药水平与其他细菌物种进行直接比较。在此,我们描述一种确定[具体细菌名称]对抗菌剂敏感性的程序,该程序可与在尽可能接近相同实验条件下测试的参考生物体进行比较。简而言之,通过测量在存在和不存在不同抗菌剂浓度的情况下捕食者/猎物混合培养物在600nm处吸光度的降低来确定[具体细菌名称]的最低抑菌浓度(MIC)值。值得注意的是,使用不同条件、猎物细菌和/或抗菌剂,该方法可进行修改以获得其他捕食性细菌的抗菌MIC值。