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Physiological state of bacteria growing in meat during cold storage: a molecular approach

机译:冷藏过程中肉类中细菌生长的生理状态:一种分子方法

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Bacteria growing on meat are subjected to specific life conditions (nutrient, pH, salt, temperature) which differ drastically from typical laboratory procedures on synthetic media, so physiological and molecular adjustments are expected. This study was conducted to determine the behaviour of bacteria when transferred from a rich-liquid medium to a solid food matrix as it is the case during microbial process validation. Planktonic Escherichia coli cultured in Brain Heart Infusion (BHI) to exponential and stationary growth phases were inoculated onto aseptically prepared ground beef meat and stored at 5°C for 12 days. No growth was observed during storage and time-course expression of sigma factors (encoded by rpoD, E, H, S) and of genes under their transcriptional control was established by RT-PCR. Overall, gene expression profiles were more stable in stationary phase bacteria. After transfer onto refrigerated meat, rpoD mRNA levels increased significantly in exponential phase bacteria (P = 0.0005) but not in stationary phase ones (P = 0.1). The rpoS mRNA levels were not significantly changed whatever the physiological state of the inoculum (P > 0.1) and a decrease of gadA transcription, controlled by σ~s, were only observed after 12 days of cold storage in exponential phase bacteria (P = 0.03). Surprisingly, activation of σ~H was only observed after more than 1 day of storage. Although the sigma factors expression profiles indicate that bacteria quickly adapt, cells do not seem to perceive a real stress after transfer from a rich medium to the cold meat environment.
机译:在肉上生长的细菌要经受特定的生活条件(营养,pH,盐,温度),这些条件与合成介质上的典型实验室程序大不相同,因此需要进行生理和分子调节。进行这项研究是为了确定细菌从丰富液体培养基转移到固体食物基质中时的行为,就像在微生物过程验证中一样。将在脑心浸液(BHI)中培养到指数生长期的浮游大肠杆菌接种到无菌制备的牛肉末上,并在5°C下保存12天。在储存期间未观察到生长,并且通过rt-PCR建立了sigma因子(由rpoD,E,H,S编码)和其转录控制下的基因随时间变化的表达。总体而言,基因表达谱在固定相细菌中更稳定。转移到冷藏肉中后,指数相细菌中的rpoD mRNA水平显着增加(P = 0.0005),而在固定相细菌中则没有(r = 0.1)。无论接种物的生理状态如何,rpoS mRNA水平都没有显着变化(P> 0.1),并且仅在指数期细菌中冷藏12天后才观察到受σ〜s控制的gadA转录下降(P = 0.03 )。出乎意料的是,仅在储存超过1天后才观察到σ〜H的活化。尽管sigma因子的表达谱表明细菌迅速适应,但从丰富的培养基转移到冷肉环境后,细胞似乎并没有感受到真正的压力。

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