Viability and Biomass of <Emphasis Type="Italic">Micrococcus luteus</Emphasis> DE2008 at Different Salinity Concentrations Determined by Specific Fluorochromes and CLSM-Image Analysis |
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Authors: | Zully M Puyen Eduard Villagrasa Juan Maldonado Isabel Esteve Antonio Solé |
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Institution: | 1.Department of Genetics and Microbiology,Universitat Autònoma de Barcelona,Barcelona,Spain |
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Abstract: | In previous studies, our group developed a method based on Confocal Laser Scanning Microscopy and Image Analysis (CLSM-IA)
to analyze the diversity and biomass of cyanobacteria in microbial mats. However, this method cannot be applied to heterotrophic
microorganisms, as these do not have autofluorescence. In this article, we present a method that combines CLSM-IA and Hoechst
33342 and SYTOX Green fluorochromes (FLU-CLSM-IA) to determine the viability and biomass of Micrococcus luteus DE2008, isolated from a saline microbial mat (Ebro Delta, Tarragona, Spain). The method has been applied to assess the effect
of salinity on this microorganism. A reduction in viability and biomass (live cells) was observed as the salt concentration
increases. The largest effect was at 100‰ NaCl with a cell death of 27.25% and a decrease in total and individual biomass
of 39.75 and 0.009 mgC/cm3, respectively, both with respect to optimal growth (10 ‰ NaCl). On the other hand, another important contribution of this
article was that combining the FLU-CLSM-IA results with those achieved by plate counts enabled us to determine, for first
time, the viability and the total biomass of the “dormant cells” (66.75% of viability and 40.59 mgC/cm3 of total biomass at 100‰ NaCl). FLU-CLSM-IA is an efficient, fast, and reliable method for making a total count of cells
at pixel level, including the dormant cells, to evaluate the viability and the biomass of a hetetrophic microorganism, M. luteus DE2008. |
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