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

发现基金研究的第一个方面将是根据密切相关的细菌菌株的傅里叶变换红外（FTIR）光谱、高分辨率魔角旋转（HRMAS）核磁共振和质谱的差异，探索它们之间的区别性质，重点是它们在微生物分型、爆发检测和生物标志物发现方面的应用。在我们的工作中，这些能力正在被用于开发快速鉴定耐抗生素细菌的方法，如耐甲氧西林金黄色葡萄球菌（MRSA）和耐万古霉素肠球菌（VRE），这是医疗保健相关感染的最普遍原因之一，以及鉴定新出现的多重耐药临床酵母菌分离物，如假丝酵母。这项工作是通过与加拿大卫生部、魁北克临床监测机构和魁北克各地多家医院的合作研究进行的。关于食源性疾病，正在与加拿大卫生部和加拿大食品检验局（CFIA）的研究人员合作，旨在通过全基因组测序和FTIR光谱研究致病性大肠杆菌病原菌（与食源性疾病相关）的遗传变异对其毒力的影响。我们的工作首先需要获取来自微生物的大量光谱数据集，用于创建包含大量FTIR， 1H和31P HRMAS NMR和质谱（MS）谱的光谱数据库，并通过应用特征选择算法结合多元统计分析技术提取歧视性光谱信息。提出的研究的一个主要目的是制定新的策略，以阐明特定细菌菌株之间的光谱差异背后的特定生化差异。预计本研究的结果将促进生物标志物的发现，不仅在我们的微生物鉴定和分类工作中，而且在快速发展的微生物分型领域，并可能有助于开发新的抗生素，以治疗不断上升的抗生素医院感染水平，导致医院爆发。该研究的第二个方面将集中于探索蛋白质与选定的多糖结合，作为可生物降解薄膜的组成部分，并在薄膜中嵌入天然抗菌剂，以延缓新鲜和最低限度加工食品的微生物生长。本研究主要是利用红外成像显微镜研究天然抗菌剂在生物降解膜中的分布和迁移速率，以提高其在活性包装中的潜在应用。

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.