Intense pulsed light (IPL) is becoming a new technical platform for disinfecting food against pathogenic bacteria. Metabolic changes are deemed to occur in bacteria as either the causes or the consequences of IPL-elicited bactericidal and bacteriostatic effects. However, little is known about the influences of IPL on bacterial metabolome. In this study, the IPL treatment was applied to E. coli K-12 for 0–20 s, leading to time- and dose-dependent reductions in colony-forming units (CFU) and morphological changes. Both membrane lipids and cytoplasmic metabolites of the control and IPL-treated E. coli were examined by the liquid chromatography–mass spectrometry (LC-MS)-based metabolomic fingerprinting. The results from multivariate modeling and marker identification indicate that the metabolites in electron transport chain (ETC), redox response, glycolysis, amino acid, and nucleotide metabolism were selectively affected by the IPL treatments. The time courses and scales of these metabolic changes, together with the biochemical connections among them, revealed a cascade of events that might be initiated by the degradation of quinone electron carriers and then followed by oxidative stress, disruption of intermediary metabolism, nucleotide degradation, and morphological changes. Therefore, the degradations of membrane quinones, especially the rapid depletion of menaquinone-8 (MK-8), can be considered as a triggering event in the IPL-elicited metabolic changes in E. coli.
Salmonella and Cronobacter are two bacteria of concern in powdered food ingredients with low water activity, due to their ability to remain viable for long periods of time. There is great interest in studying the survival of these bacteria in powdered foods, but discrepancies have been reported between broth-grown and lawn-grown bacterial cells and their thermal resistance and desiccation tolerance once inoculated onto powdered foods. The purpose of this study was to evaluate three different powdered food inoculation methods, two broth-grown and one lawn-grown. To evaluate these methods on three types of powdered food matrices, Salmonella enterica serovar Typhimurium LT2 (ATCC 700720), Salmonella surrogate Enterococcus faecium (NRRL B-2354), and Cronobacter sakazakii (ATCC 29544) were inoculated onto nonfat dry milk powder, organic soy flour, and all-purpose flour using one of the three previously developed inoculation methods. In the first broth-grown method, labeled broth-grown pelletized inoculation, a bacterial cell pellet was added to powdered foods directly and mixed with a sterile wooden stick. The second broth-grown method, labeled broth-grown spray inoculation, used a chromatography reagent sprayer to spray the bacterial cell suspension onto the powdered foods. The third inoculation method, lawn-grown liquid inoculation, made use of a spot inoculation and a stomacher to incorporate each bacterium into the powdered foods. Results indicated that the method of inoculation of each powder impacted repeatability and bacteria survivability postequilibration (4 to 6 days). Broth-grown spray inoculation, regardless of the powder and bacterium, resulted in the highest log reduction, with an average ∼1-log CFU/g reduction following equilibration. Broth-grown pelletized inoculation resulted in the second-highest log reduction (∼0.79 log CFU/g), and finally, lawn-grown liquid inoculation was the most stable inoculation method of the three, with ∼0.52-log CFU/g reduction. Overall, the results from this inoculation study demonstrate that inoculation methodologies impact the desiccation tolerance and homogeneity of C. sakazakii, E. faecium, and Salmonella Typhimurium LT2.
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