BIOLUMINOMETRIC ASSAY OF CELL ATP DETECTION UNDER STRESS CONDITIONS AND IN THE ASSESSMENT OF CLEANNESS OF THE FOOD PRODUCING COMPANY

M. Požgajová, H. Ďúranová
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Abstract

Detection and tracking of microorganism’s behavior in natural environments and under stressed conditions have been nowadays accelerated by the requirement for the risk assessment associated with environmental contamination. Luminescence-based techniques, for they cost efficiency, fastness, reliability and reproducibility are particularly appropriate for such studies. We used bioluminescence sensing assay to determine ATP levels in the yeast Schizosaccharomyce pombe. Determination of adenosine concentration per cell was assessed in the yeast cells counted prior to the experiment. To test the methodology, model organism was exposed to life-threatening concentration of heavy metals cadmium and nickel for three hours. Afterwards, changes in the ATP levels were detected and compared to the control. Strikingly, our results revealed that such conditions lead to considerable increase in the production of ATP by mitochondria compared to untreated cells. Normal level of ATP in the control increased from the basal 9.92x10-16 M to 2.90x10-15 M after Cd exposure and to 7.58x10-15 M upon Ni treatment. The methodology was subsequently used for assessment of cleanness of margarine producing company devices after regular sanitation procedure. Detection of only negligible amount of the ATP signal confirmed good cleaning strategy in the tested food processing company.
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胁迫条件下细胞atp检测及食品生产企业清洁度评价的生物荧光分析
环境污染风险评估的要求加快了对自然环境和应激条件下微生物行为的检测和跟踪。基于发光的技术,因为它们具有成本效益、快速、可靠和可重复性,特别适合于这类研究。采用生物发光传感法测定了酵母裂糖酵母(Schizosaccharomyce pombe)中ATP的含量。每个细胞腺苷浓度的测定在实验前计数的酵母细胞中进行评估。为了验证该方法,将模式生物暴露在危及生命的重金属镉和镍中三个小时。之后,检测ATP水平的变化,并与对照组进行比较。引人注目的是,我们的结果显示,与未经处理的细胞相比,这种情况导致线粒体产生ATP的数量显著增加。对照的正常ATP水平在Cd处理后从基础的9.92x10-16 M上升到2.90x10-15 M,在Ni处理后上升到7.58x10-15 M。该方法随后被用于人造黄油生产公司设备在常规卫生程序后的清洁度评估。仅检测到可忽略不计的ATP信号,证实了被测食品加工公司的良好清洁策略。
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