Short-term deleterious effects of standard isolation and cultivation methods on new tropical freshwater microalgae strains

M Magdalena Aray-Andrade; Miguel I Uyaguari-Diaz; J Rafael Bermúdez

doi:10.7717/peerj.5143

Short-term deleterious effects of standard isolation and cultivation methods on new tropical freshwater microalgae strains

PeerJ. 2018 Jul 19:6:e5143. doi: 10.7717/peerj.5143. eCollection 2018.

Authors

M Magdalena Aray-Andrade^#^{1

2

3}, Miguel I Uyaguari-Diaz^{4

5}, J Rafael Bermúdez^#^{1

6}

Affiliations

¹ ESPOL Polytechnic University, Escuela Superior Politécnica del Litoral, ESPOL, Plankton Laboratory, Facultad de Ingenería Marítima, Ciencias Biológicas, Oceánicas y Recursos Naturales, Campus Gustavo Galindo, Guayaquil, Ecuador.
² Joint Postgraduate VLIR NETWORK Master Program in Applied Biosciences, Biodiscovery, ESPOL Polytechnic University, Guayaquil, Guayas, Ecuador.
³ Escuela de Medicina, Universidad Espíritu Santo-Ecuador, Samborondón, Guayas, Ecuador.
⁴ British Columbia Centre for Disease Control Public Health Laboratory, Vancouver, BC, Canada.
⁵ Department of Pathology and Laboratory Medicine, University of British Columbia, Vancouver, BC, Canada.
⁶ Galapagos Marine Research and Exploration, GMaRE. Joint ESPOL-CDF program, Charles Darwin Research Station, Galapagos Islands, Ecuador.

^# Contributed equally.

Abstract

Algae with potential biotechnological applications in different industries are commonly isolated from the environment in order to obtain pure (axenic) stocks that can be safely stored for long periods of time. To obtain axenic cultures, antibiotics are frequently employed, and cryopreservation is applied to preserve standing stocks. However, many of these now standard methods were developed using strains derived from pristine to near-pristine environments and cold to temperate regions. The potential effect of the said methods on the life cycle and biochemical profile of algae isolates from hyper-eutrophic and constant high-temperature tropical regions is not well understood. These effects could potentially render them unsuitable for their intended biotechnological application. In this study, we conducted a genetic characterization (18S rRNA) and evaluated the effect of purification (the use of the antibiotic chloramphenicol, CAP) and cryopreservation (dimethyl sulfoxide; DMSO-sucrose mix and glycerol) on the growth rate and lipid content of three new tropical freshwater algal isolates: Chorella sp. M2, Chlorella sp. M6, and Scenedesmus sp. R3, obtained from the Ecuadorian coast. The genetic and morphological characterization revealed a clear discrimination between these strains. All strains cultured with CAP exhibited a lower growth rate. Subsequent to cryopreservation, Chorella sp. M2, Chlorella sp. M6, and Scenedesmus sp. R3 presented no significant difference in growth rate between the cryopreservants. Further, a significantly higher lipid content was observed in the biomass cryopreserved with glycerol in relation to the DMSO-sucrose, with Chorella sp. M2 and Chlorella sp. M6 having twice as much as they had in the first treatment. These results highlight the relevance of selecting an appropriate method for storage, as the materials used can affect the biological performance of different tropical species, although it is still to be determined if the effects observed in this study are long lasting in subsequent cultures of these algae.

Keywords: Antibiotics; Cryopreservation; Growth rate; Lipids; Microalgae.

Grants and funding

This project was funded by the Dean of Research at ESPOL, by the grant G1-DI-2015. This project was executed within the Join postgraduate VLIR NETWORK Master Program in Applied Biosciences. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.