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Flow cytometric analysis to evaluate physiological alterations in herbicide-exposed Chlamydomonas moewusii cells

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Abstract

Investigation of herbicide toxicology in non-target aquatic primary producers such as microalgae is of great importance from an ecological point of view. In order to study the toxicity of the widely used herbicide paraquat on freshwater green microalga Chlamydomonas moewusii, physiological changes associated with 96 h-exposures to this pollutant were monitored using flow cytometry (FCM) technique. Intracellular reactive oxygen species concentration, cytoplasmic membrane potential, metabolic activity and cell protein content were monitored to evaluate the toxicological impact of paraquat on algal physiology. Results showed that herbicide paraquat induced oxidative stress in C. moewusii cells, as it indicated the increase of both superoxide anion and hydrogen peroxide levels observed in non-chlorotic cells of cultures exposed to increasing herbicide concentrations. Furthermore, a progressive increase in the percentage of depolarised cells and a decrease in the metabolic activity level were observed in response to paraquat when non-chlorotic cells were analysed. Chlorotic cells were probably non-viable cells, based on the cytoplasmic membrane depolarisation, its metabolically non-active state and its drastically reduced protein content. In view of the obtained results, we have concluded that a range of significant physiological alterations, detected by flow cytometry, occur when C. moewusii, an ubiquitous microalga in freshwater environments, is challenged with environmentally relevant concentrations of the herbicide paraquat.

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Acknowledgments

This work was supported by research project from the Spanish Government’s Ministerio de Educación e Innovación (CGL2004-02037). R. Prado gratefully acknowledges a F.P.U. fellowship from the Spanish Ministerio de Educación y Ciencia. The authors wish to thank Gerardo Fernández (SAI-UDC) his willingness to carry out the determination of paraquat in water, and also thank the suggestions made by reviewers to improve this article.

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  1. Laboratorio de Microbiología, Facultad de Ciencias, Universidad de A Coruña, Campus de A Zapateira s/n, 15008 A, Coruña, Spain

    Raquel Prado, Carmen Rioboo, Concepción Herrero, Paula Suárez-Bregua & Ángeles Cid

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  1. Raquel Prado
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Correspondence to Ángeles Cid.

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Prado, R., Rioboo, C., Herrero, C. et al. Flow cytometric analysis to evaluate physiological alterations in herbicide-exposed Chlamydomonas moewusii cells. Ecotoxicology 21, 409–420 (2012). https://doi.org/10.1007/s10646-011-0801-3

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