title: High-throughput screening of microbial adaptation to environmental stress.
abstract: We developed a microwell plate, high-throughput, screening method aimed at quantitating the tolerance of a panel of Gram-positive and Gram-negative bacteria to metals (Frankia sp., Escherichia coli, Cupriavidus metallidurans, Rhizobium leguminosarum, and Streptomyces scabies). Microbial viability was quantified using MTS; a tetrazolium salt converted to a water-soluble formazan through microbial reduction. In this paper, we present the stepwise development of the method, highlighting the main elements underlying its reliability, and compare results obtained with literature. We conclude the method is well suited to efficiently screen bacteria, including those that are filamentous and slow-growing, without the need for large amounts of inoculum which may not always be available. The method allows testing of compound gradients with sufficient replicates to generate statistically robust results, and is transposable to other types of cell proliferation assays such as those for antimicrobial susceptibility, and chemoresistance.
authors.affiliation: Centre d'étude et de valorisation de la diversité microbienne, Département de Biologie, Université de Sherbrooke, Sherbrooke, QC, Canada.
authors.forename: Pier-Anne
authors.initials: PA
authors.lastname: Bélanger
authors.forename: Julie
authors.initials: J
authors.lastname: Beaudin
authors.forename: Sébastien
authors.initials: S
authors.lastname: Roy
mesh_terms: D000220:Adaptation, Biological; D029661:Alnus / Q000382:microbiology; D004777:Environment; D005562:Formazans / Q000378:metabolism; D040161:Frankia / Q000302:isolation & purification* / Q000502:physiology*; D057166:High-Throughput Screening Assays / Q000379:methods*; D008670:Metals / Q000378:metabolism; D050296:Microbial Viability; D010084:Oxidation-Reduction; D013312:Stress, Physiological; D013778:Tetrazolium Salts / Q000378:metabolism
chemical_list: D005562:Formazans; D008670:Metals; D013778:Tetrazolium Salts