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J Appl Phycol (2016) 28:105112 DOI 10.1007/s10811-015-0585-6
Rapid method for the assessment of cell lysis in microalgae cultures
J. J. Gallardo-Rodrguez1 & L. Lpez-Rosales1 & A. Snchez-Mirn1 &
F. Garca-Camacho1 & E. Molina-Grima1
Received: 7 January 2015 /Revised and accepted: 6 April 2015 /Published online: 17 April 2015 # Springer Science+Business Media Dordrecht 2015
Abstract A solid understanding of the effect of hydrodynamic forces encountered by microalgae in bioprocesses would benefit existing bioprocesses, eventually allowing an increase in their productivity. For this purpose, a sensitive method able to quantify cell lysis is crucial. Most of the available protocols and methods intended for this purpose were developed for animal or insect cells. In the case of microalgae, the commercial kits tested were unable to determine the cell lysis extension. The method proposed here was developed to relate the release of a cytoplasmic component (enzyme lactate dehydrogenase (LDH)) with cell lysis by measuring the NADH (reduced form of nicotinamide cofactor adenine dinucleotide) produced by LDH. Although different commercial kits based on similar processes are available, they are more complicated to use and not applicable to microalgae nor when longer-term tests are to be performed.
Keywords Cell lysis . Viability . LDH . Microalgae .
Dinoflagellate
Introduction
Microalgae are used in different bioprocesses for biofuel production (Chisti 2007), bioremediation (Kraemer et al. 2004; de-Bashan and Bashan 2010; Doria et al. 2012), and CO2 capture (Aishvarya et al. 2012; Gonzlez-Lpez et al. 2012)
and can provide numerous high-value products (Singh et al.
2005; Camacho et al. 2007; Gallardo-Rodrguez et al. 2012). Pneumatically agitated photobioreactors have proved useful for microalgal culture, but certain algae are damaged by hydrodynamic stress in these systems (Gudin and Chaumont 1991; Jaouen et al. 1999; Chae et al. 2006; Camacho et al. 2011). Shear sensitivity of algal cultures can severely restrict attainable productivity in photobioreactors (Gallardo Rodrguez et al. 2009).
The most common methods for quantifying cell viability are based on the ability of the membrane of viable cells to exclude certain dyes such as trypan blue or propidium iodide (Moore et al. 1998). Viable cells are not stained with such dyes. In contrast, dead or non-viable cells, which have lost the integrity of the cell membrane, allow the passage of these dyes to the...