Using flow cytometry to monitor the stress response of yeast and microalgae populations in mixed cultures developed in b
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Using flow cytometry to monitor the stress response of yeast and microalgae populations in mixed cultures developed in brewery effluents Carla Dias 1 & Luísa Gouveia 1 & José A. L. Santos 2,3 & Alberto Reis 1 & Teresa Lopes da Silva 1 Received: 17 March 2020 / Revised and accepted: 18 August 2020 # Springer Nature B.V. 2020
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. Keywords Flow cytometry . Yeast . Microalgae . Mixed culture . Stress response
Introduction Recently, microalgae/yeast co-cultures have been considered a promising approach to improve microbial biomass productivity, cell oil content, and lipid productivity, presenting several advantages over pure cultures. In addition, when using effluents as feedstock, mixed cultures allow better nutrient removal rates (Dias et al. 2019). Yeast and microalgae mixed cultures take advantage of the complementary heterotrophic/ autotrophic nutritional modes: the microalgae consume carbon dioxide and produce oxygen, through photosynthesis, which is consumed by the yeast, which in turn supply carbon dioxide through respiration, to be consumed by the microalgae. It has been reported that yeast and microalgae co-cultures allow both
* Teresa Lopes da Silva [email protected] 1
Laboratório Nacional de Energia e Geologia, I.P., Unidade de Bioenergia e Biorrefinarias, Estrada do Paço do Lumiar, 22 1649-038 Lisboa, Portugal
2
Departamento de Bioengenharia, Instituto Superior Técnico, Universidade de Lisboa, Avenida Rovisco Pais, 1049-001 Lisboa, Portugal
3
IBB, Institute for Biotechnology and Bioengineering, Avenida Rovisco Pais, 1049-001 Lisboa, Portugal
microorganisms to produce higher biomass yields (Cheirsilp et al. 2011; Santos et al. 2013). Likewise, there are also metabolites exchanges and pH medium adjustment that are benefic for both microorganisms (Dias et al. 2019). To make the process economically sustainable, different low-cost substrates have been used for microalgal lipid production. The nutrients from the effluents and other low-cost substrates usually can fulfill the nutritional requirements of the microorganisms; howev
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