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Identification and quantification of suspended algae and bacteria populations using flow cytometry: applications for algae biofuel and biochemical growth systems

Abstract

Species diversity in algae biofuel and biochemical culturing systems can affect yield; in many large-scale algae growth systems, it is not practical to maintain a monoculture. To better understand and monitor these complex systems, techniques are required which can quickly and effectively quantify the species distribution and overall growth of a mixed microbial community in suspension. A flow cytometric method has been developed which can be used to differentiate populations of three Chlorophyta species, one diatom species, cyanobacteria, and heterotrophic bacteria according to their fluorescence and morphology. The nucleic acid stain SYTO9 was used to discriminate species with similar natural autofluorescence and to identify heterotrophic bacteria. Absolute cell enumeration was performed with counting beads and validated with a hemocytometer. Species identification was validated by analyzing known mixtures of axenic cell cultures. The utility of the method was demonstrated by studying the effect of light intensity on species succession, growth, and biomass accumulation in small algae growth systems over 22 days. Flow cytometric analysis, augmented with SYTO9 stain and counting beads, can be utilized to monitor algae biofuel and biochemical growth systems involving multiple species. This method allows for monitoring of contamination, succession, and overall growth in both natural and intentionally created microbial communities.

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Acknowledgments

The authors would like to thank the Natural Science and Engineering Research Council of Canada (NSERC) for financial support in the form of a strategic grant, and a Canadian Graduate Scholarship (CGS D). Additional thanks to the Division of Engineering Science at the University of Toronto for supplementary financial support. Lastly, special acknowledgement to Dionne White at the University of Toronto Faculty of Medicine Flow Cytometry Facility for her guidance with the flow cytometer.

Conflict of interest

The authors declare that they have no conflict of interest.

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Correspondence to D. G. Allen.

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Peniuk, G.T., Schnurr, P.J. & Allen, D.G. Identification and quantification of suspended algae and bacteria populations using flow cytometry: applications for algae biofuel and biochemical growth systems. J Appl Phycol 28, 95–104 (2016). https://doi.org/10.1007/s10811-015-0569-6

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  • DOI: https://doi.org/10.1007/s10811-015-0569-6

Keywords

  • Phytoplankton
  • Flow cytometry
  • Biofuels
  • Bacteria
  • Quantification
  • SYTO9 dye