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在连续运行的搅拌-管式生物反应器系统中研究大肠杆菌生物膜的形成及其对 T4 噬菌体的敏感性。

E. coli biofilm formation and its susceptibility towards T4 bacteriophages studied in a continuously operating mixing - tubular bioreactor system.

机构信息

Faculty of Chemistry and Chemical Technology, University of Ljubljana, Večna pot, 113, Ljubljana, Slovenia.

COBIK, Mirce 21, 5270, Ajdovščina, Slovenia.

出版信息

Microb Biotechnol. 2022 Sep;15(9):2450-2463. doi: 10.1111/1751-7915.14079. Epub 2022 May 31.

Abstract

A system consisting of a connected mixed and tubular bioreactor was designed to study bacterial biofilm formation and the effect of its exposure to bacteriophages under different experimental conditions. The bacterial biofilm inside silicone tubular bioreactor was formed during the continuous pumping of bacterial cells at a constant physiological state for 2 h and subsequent washing with a buffer for 24 h. Monitoring bacterial and bacteriophage concentration along the tubular bioreactor was performed via a piercing method. The presence of biofilm and planktonic cells was demonstrated by combining the piercing method, measurement of planktonic cell concentration at the tubular bioreactor outlet, and optical microscopy. The planktonic cell formation rate was found to be 8.95 × 10  h and increased approximately four-fold (4×) after biofilm exposure to an LB medium. Exposure of bacterial biofilm to bacteriophages in the LB medium resulted in a rapid decrease of biofilm and planktonic cell concentration, to below the detection limit within < 2 h. When bacteriophages were supplied in the buffer, only a moderate decrease in the concentration of both bacterial cell types was observed. After biofilm washing with buffer to remove unadsorbed bacteriophages, its exposure to the LB medium (without bacteriophages) resulted in a rapid decrease in bacterial concentration: again below the detection limit in < 2 h.

摘要

设计了一个由连通的混合和管状生物反应器组成的系统,以在不同实验条件下研究细菌生物膜的形成及其暴露于噬菌体的影响。在连续泵送细菌细胞以恒定生理状态 2 小时并随后用缓冲液洗涤 24 小时的过程中,在硅酮管状生物反应器内部形成细菌生物膜。通过穿孔法监测管状生物反应器内的细菌和噬菌体浓度。通过结合穿孔法、管状生物反应器出口处浮游细胞浓度的测量和光学显微镜观察,证明了生物膜和浮游细胞的存在。发现浮游细胞的形成速率为 8.95×10 h,并且在生物膜暴露于 LB 培养基后增加了约四倍(4×)。将细菌生物膜暴露于 LB 培养基中的噬菌体导致生物膜和浮游细胞浓度迅速下降,在<2 小时内降至检测限以下。当噬菌体在缓冲液中供应时,仅观察到两种细菌细胞类型的浓度适度下降。在用缓冲液洗涤生物膜以去除未吸附的噬菌体后,将其暴露于 LB 培养基(不含噬菌体)导致细菌浓度迅速下降:再次在<2 小时内降至检测限以下。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/cecb/9437887/cd39cf6ff70b/MBT2-15-2450-g002.jpg

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