Rapid detection and respirometric profiling of aerobic bacteria on panels of selective media

Abstract

To evaluate high-throughput optical oxygen microrespirometry for selective detection and predictive identification of aerobic bacteria. Using GreenLight probe, standard 384-well plates and time-resolved fluorescence reader, a representative panel of 16 partially selective media and 9 aerobic bacteria (Escherichia coli, Bacillus cereus, Staphylococcus aureus, Campylobacter jejuni, Yersinia enterocolitica, Pseudomonas aeruginosa, Streptococcus pyogenes, Salmonella typhimurium and Listeria innocua) were analysed. For each medium, bacterial strain and dilution, growth profiles were recorded, from which calibrations, doubling/generation times and growth patterns in different media were determined. Analytical performance, selectivity and general usability of the method were assessed, and mixed bacterial cultures were analysed. The microrespirometry platform facilitates simple, real-time detection and predictive identification of aerobic bacteria by looking at the patterns of their growth and respiration in several media and determining their growth and doubling times. The new screening method can be used for routine microbiological analysis and testing of aerobic bacterial cultures as well as complex food, environmental and clinical samples.