Exploration of new approaches to microbial typing and phenotypic characterization based on Fourier transform infrared and high-resolution magic-angle spinning NMR spectroscopy and the development of active packaging employing natural antimicrobials

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The first facet of the proposed Discovery Grant research will be directed toward the exploration of the discriminatory nature between closely related bacterial strains based on differences in their Fourier transform infrared (FTIR) spectra, high-resolution magic angle spinning (HRMAS) NMR and mass spectrometry with a focus on their application to microbial typing, outbreak detection and biomarker discovery. In our work, these capabilities are being exploited to develop rapid methods for identification of antibiotic-resistant bacteria such as methicillin-resistant Staphylococcus aureus (MRSA) and vancomycin-resistant enterococci (VRE), which are among the most prevalent causes of healthcare-associated infections, and identification of emerging multi-drug resistant clinical yeast isolates such as Candida auris. This work is being conducted through collaborative studies with Health Canada, the Quebec clinical surveillance agency and multiple hospitals across Quebec. With regards to foodborne illness, ongoing efforts, in collaboration with researchers at Health Canada and the Canadian Food Inspection Agency (CFIA), is aimed at studying the effect of genetic variability of pathogenic E. coli pathotypes (associated with foodborne illness) to its virulence by whole genome sequencing and FTIR spectroscopy. Our work initially entails acquisition of a large spectral datasets from of microorganisms for the creation of spectral databases containing large numbers of FTIR, 1H and 31P HRMAS NMR and mass spectrometry (MS) spectra of well-characterized strains and extracting the discriminatory spectral information by applying feature-selection algorithms in conjunction with multivariate statistical analysis techniques. A major objective of the proposed research is to develop new strategies directed toward elucidation of the specific biochemical differences underlying the spectral differences among particular bacterial strains. It is anticipated that the results of the proposed research will advance biomarker discovery not only in our work on microbial identification and classification but also in the rapidly evolving field of microbial typing and may aid in the development of new antibiotics that can treat the rising levels of antibiotic nosocomial infections resulting in hospital outbreaks. The second facet of the proposed research will focus on the exploration of proteins in combination of selected polysaccharides as components of biodegradable films and embedding the films with natural antimicrobials to retard microbial growth of fresh and minimally processed foods. The proposed research focuses on the application of infrared imaging microscopy to study the distribution, migration rates of natural antimicrobials in biodegradable films in relation to enhancing their potential use in active packaging. *****