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微量热法:跟踪异形生物对梨形四膜虫毒性的强大而新颖的工具。

Microcalorimetry: a powerful and original tool for tracking the toxicity of a xenobiotic on Tetrahymena pyriformis.

机构信息

Clermont Université, Université Blaise Pascal, Laboratoire "Microorganismes : Génome et Environnement", BP10448, F-63000 Clermont-Ferrand, France; CNRS, UMR6023, LMGE BP80026, F-63171 Aubière, France.

出版信息

Ecotoxicol Environ Saf. 2013 Dec;98:88-94. doi: 10.1016/j.ecoenv.2013.09.019. Epub 2013 Oct 20.

Abstract

Fighting against water pollution requires the ability to evaluate the toxicity of pollutants such as herbicides. Tetrahymena pyriformis are ubiquitous ciliated protozoans commonly used in ecotoxicological research. Microcalorimetry can be used in many biological investigations as a universal, non-destructive and highly sensitive tool that provides a continuous real-time monitoring of the metabolic activity. This technique based on the thermal power output was applied to evaluate the influence of herbicide diuron on cultures of T. pyriformis. The heat flux produced upon addition of 0, 3.5, 7.0, 14.0, 28.0, and 56.0 µg mL⁻¹ of diuron was monitored. The biomass change during the growth was also determined by flow cytometry. The results confirmed that the growth of T. pyriformis is progressively inhibited as the concentration of diuron increases and revealed that the state of the living system is severely altered at a concentration of 56.0 µg mL⁻¹. The IC₅₀ was estimated at 13.8 µg mL⁻¹ by microcalorimetry and at 18.6 µg mL⁻¹ by flow cytometry. It was shown that microcalorimetry is not only a very effective tool for the determination of the growth rate constant but that it is also a valuable probe for a rapid detection of the metabolic perturbations and, in ultimate cases, of the critical alterations of the living system under the action of a toxic agent.

摘要

防治水污染需要能够评估除草剂等污染物的毒性。梨形四膜虫是普遍存在的有纤毛原生动物,常用于生态毒理学研究。微量热法可以在许多生物学研究中用作通用的、非破坏性的和高灵敏度的工具,提供对代谢活性的连续实时监测。这项基于热功率输出的技术被应用于评估除草剂敌草隆对梨形四膜虫培养物的影响。监测添加 0、3.5、7.0、14.0、28.0 和 56.0 µg mL⁻¹敌草隆时产生的热通量。通过流式细胞术还确定了生长过程中的生物量变化。结果证实,随着敌草隆浓度的增加,梨形四膜虫的生长逐渐受到抑制,并表明在 56.0 µg mL⁻¹的浓度下,生物系统的状态严重改变。用微量热法和流式细胞术分别估计 IC₅₀为 13.8 µg mL⁻¹和 18.6 µg mL⁻¹。结果表明,微量热法不仅是确定生长速率常数的非常有效的工具,而且还是快速检测代谢扰动的有价值的探针,在极端情况下,还可以检测在有毒剂作用下生物系统的临界变化。

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