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Using flow cytometry to monitor the stress response of yeast and microalgae populations in mixed cultures developed in brewery effluents

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Abstract

Recently, yeast and microalgae mixed cultures have been widely used in biological effluent treatments and biofuel production because such cultures show many advantages over pure cultures. However, industrial effluents often contain toxic compounds; therefore, it is important to evaluate the cell stress response when growing in such conditions during the mixed culture development. In this work, flow cytometry (FC) was used to differentiate Rhodosporidium toruloides cells from Tetradesmus obliquus cells, based on their size, internal complexity, and chlorophyll content. FC coupled with SYTOX Green and CFDA fluorochromes was also used to characterize the cell stress response of R. toruloides and T. obliquus individual cells in a mixed culture. This work describes, for the first time, a simple and easy method to monitor individual stress response of R. toruloides and T. obliquus cells growing in mixed cultures on brewery effluents, using FC coupled with fluorescent dyes.

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Acknowledgments

The authors acknowledge Sociedade Central de Cervejas e Bebidas, S.A (Vialonga, Portugal) and Dr. António Sérgio Marques, from Sidul Campany (Alhandra, Portugal), for the kindly supply of brewery effluent and sugarcane molasses, respectively.

Funding

Carla Dias PhD scholarship is sponsored by FCT (Fundação para a Ciência e Tecnologia), Portugal (SFRH/BD/117355/2016). The authors also thank the Biomass and Bioenergy Research Infrastructure (BBRI)-LISBOA-01-0145-FEDER-022059, which is supported by Operational Programme for Competitiveness and Internationalization (PORTUGAL2020), by Lisbon Portugal Regional Operational Programme (Lisboa 2020) and by North Portugal Regional Operational Programme (Norte 2020) under the Portugal 2020 Partnership Agreement, through the European Regional Development Fund (ERDF).

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Authors and Affiliations

  1. Laboratório Nacional de Energia e Geologia, I.P., Unidade de Bioenergia e Biorrefinarias, Estrada do Paço do Lumiar, 22 1649-038, Lisboa, Portugal

    Carla Dias, Luísa Gouveia, Alberto Reis & Teresa Lopes da Silva

  2. Departamento de Bioengenharia, Instituto Superior Técnico, Universidade de Lisboa, Avenida Rovisco Pais, 1049-001, Lisboa, Portugal

    José A. L. Santos

  3. IBB, Institute for Biotechnology and Bioengineering, Avenida Rovisco Pais, 1049-001, Lisboa, Portugal

    José A. L. Santos

Authors
  1. Carla Dias
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  2. Luísa Gouveia
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  3. José A. L. Santos
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  4. Alberto Reis
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  5. Teresa Lopes da Silva
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Correspondence to Teresa Lopes da Silva.

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Dias, C., Gouveia, L., Santos, J.A.L. et al. Using flow cytometry to monitor the stress response of yeast and microalgae populations in mixed cultures developed in brewery effluents. J Appl Phycol 32, 3687–3701 (2020). https://doi.org/10.1007/s10811-020-02236-8

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  • DOI: https://doi.org/10.1007/s10811-020-02236-8

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