Multiwavelength Resonance Raman Characterization of the Effect of Growth Phase and Culture Medium on Bacteria

Abstract

We examine the use of multiwavelength ultraviolet (UV) resonance–Raman signatures to identify the effects of growth phase and growth medium on gram-positive and gram-negative bacteria. Escherichia coli(E. coli), Citrobacter koseri(C. koseri), Citrobacter braakii (C. braakii), and Bacillus cereus (B. cereus)were grown to logarithmic and stationary phases in nutrient broth and brain heart infusion broth. Resonance Raman spectra of bacteria were obtained at multiple wavelengths between 220 and 260 nm; a range that encompasses the resonance frequencies of cellular constituents. We find that spectra of the same bacterial species exhibit differences due to both growth condition and growth phase, but the larger differences reflect changes due to growth phase. The differences in the Raman spectra correlate with genetic differences among the species. Using a Pearson correlation based algorithm, we achieve successful identification of these bacteria in 83% of the cases.