Kheirjou Saeid, Hosseini Farzaneh, Masjedian Jazi Framarz, Siasi Torbati Elham
Department of Microbiology, North Tehran Branch, Islamic Azad University, Tehran, Iran.
Department of Microbiology, School of Medicine, Iran University of Medical Sciences, Tehran, Iran.
Rep Biochem Mol Biol. 2024 Jan;12(4):643-651. doi: 10.61186/rbmb.12.4.643.
In this study, spore-forming probiotics were employed to eradicate biofilms and the presence and expression of genes involved in stress response was examined.
Polymerase chain reaction (PCR) assay was used to detect and genes in S. ATCC 12228. Biofilm production was investigated by microtiter plate (MTP) assay. 100X minimum inhibitory concentration (MIC) of gentamycin was used to induce persister cells in planktonic and biofilm bacterial cells. The expression of , and mazF genes was assessed at different time intervals of 2, 8, and 24 h using real-time PCR assay. Then, dilutions of 1, 0.5, and 0.25 µg/ml of the supernatant of culture was used to eradicate the persister cells and the number of colonies was determined.
Persister cells of S. were formed after 7 h in planktonic and 5 h in the biofilm structure after exposure to 50 µg/ml of gentamycin. The expression of and in biofilm structure and the expression of and in persister cells were significantly higher compared to the control (p< 0.05). The number of persister cells showed a reduction of log 2.4 and log 0.8 after exposure to 1 and 0.5 µg/ml B. supernatant, respectively, but no reduction was observed at the concentration of 0.25 µg/ml.
The results showed that the supernatant of probiotics containing their secretive metabolites can be used as a novel approach to combat persister cells.
在本研究中,使用产芽孢益生菌来根除生物膜,并检测参与应激反应的基因的存在和表达。
采用聚合酶链反应(PCR)检测法检测嗜热栖热菌ATCC 12228中的基因。通过微量滴定板(MTP)检测法研究生物膜的形成。使用庆大霉素的100倍最低抑菌浓度(MIC)诱导浮游和生物膜细菌细胞中的持留菌。使用实时PCR检测法在2、8和24小时的不同时间间隔评估基因、和mazF的表达。然后,使用嗜热栖热菌培养上清液的1、0.5和0.25μg/ml稀释液来根除持留菌,并确定菌落数量。
在暴露于50μg/ml庆大霉素后,浮游状态下嗜热栖热菌的持留菌在7小时后形成,生物膜结构中的持留菌在5小时后形成。与对照组相比,生物膜结构中的基因和的表达以及持留菌中的基因和的表达显著更高(p<0.05)。分别暴露于1μg/ml和0.5μg/ml嗜热栖热菌上清液后,持留菌数量分别减少了log 2.4和log 0.8,但在0.25μg/ml浓度下未观察到减少。
结果表明,含有其分泌代谢产物的益生菌上清液可作为对抗持留菌的一种新方法。