双歧杆菌在增殖培养中存在营养充足但增殖受限的情况,因此探究其增殖过程中的分裂周期至关重要。该研究选择长双歧杆菌婴儿亚种B8762为研究对象,采用荧光探针对菌体细胞进行标记,并通过流式细胞术对该菌株在不同温度下的分裂进程及膜结构完整性进行了研究。结果表明,在10 μmol/L 荧光染料标记下,菌株表现出良好的生长状况,满足分裂过程的指标测定。此外,在不同培养条件下,各世代细胞群体的分布存在差异,表明菌体分裂过程持续进行,且各世代菌体分裂不同步,高温下“6”世代的群体数量相较于37 ℃上涨了142.31%。菌株的生理状态受到培养温度的影响较大,高温下膜结构完整性损失比例增大了 168.82%。该研究揭示了双歧杆菌增殖分裂过程中的荧光信号变化及菌体细胞活性的差异,为高活性乳酸菌的工业化生产提供了理论指导。
In the proliferation culture of Bifidobacterium, despite sufficient nutrients, the growth is found to be constrained, emphasizing the crucial exploration of its division cycle.This study focused on Bifidobacterium longum subsp.infantis B8762, employing fluorescent probes to label bacterial cells and utilizing flow cytometry to investigate the division process and membrane integrity under different temperatures.Results demonstrated that, when labelled with 10 μmol/L fluorescent dye, the strain exhibited robust growth conditions, meeting the criteria for division process assessment.Moreover, variations in the distribution of cell populations across different generations under various culture conditions indicated ongoing bacterial division, with asynchronous division among different generations.Notably, at elevated temperatures, the population of the 6th generation increased by 142.31% compared to 37 ℃.The physiological state of the strain was significantly influenced by the cultivation temperature, as evidenced by a 168.82% increase in membrane integrity loss at higher temperatures.This research unveils the fluorescent signal changes and cellular activity differences during the proliferation and division process of Bifidobacterium, providing theoretical guidance for the industrial production of highly active lactic acid bacteria.
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