Physiological response of Bacillus cereus vegetative cells to simulated food processing treatments.
Vegetative cells of the spore-former Bacillus cereus were exposed to a number of treatments commonly used in commercial food preparation or during equipment cleaning and decontamination. Treated suspensions were then analyzed for reductions (CFU per milliliter) by plate counting and changes in levels of ATP and ADP released from cells with a bioluminescence-based assay. With the use of flow cytometry (FCM), the physiological status of individual cells before and after exposure to treatments was determined by staining of control and treated cells with three pairs of physiological dyes (SYTO 9/propidium iodide, carboxyfluorescein diacetate/Hoechst 33342, and C12-resazurin/SYTOX Green). Good agreement was found between plate counting and FCM. In general, treatments giving rise to the highest count reductions also had the greatest effects on cell membrane permeability (measured with the use of propidium iodide or SYTOX Green), esterase activity (measured with carboxyfluorescein diacetate), or redox activity (C12-resazurin). FCM data demonstrated the extent of heterogeneity of vegetative cell responses to treatments in, for example, the treatment with 5% H2O2, which caused a 6-log reduction in which approximately 95% of the population was composed of membrane-damaged cells (as reflected by their permeability to SYTOX Green), whereas in treatment with 0.09% (wt/vol) potassium sorbate, which caused only a 1-log reduction, not more than 40% of cells were membrane damaged. The approaches described in this work can be applied to gain a greater understanding of bacterial responses to food control measures, generate more accurate inactivation models, or screen novel prospective food control measures.
Cronin
Ultan P
UP
Department of Life Sciences, University of Limerick, Castletroy, County Limerick, Ireland.
Wilkinson
Martin G
MG
D000244:Adenosine Diphosphate / Q000378:metabolism; D000255:Adenosine Triphosphate / Q000378:metabolism; D001409:Bacillus cereus / Q000201:enzymology / Q000254:growth & development / Q000502:physiology*; D002463:Cell Membrane Permeability / Q000187:drug effects* / Q000502:physiology; D015169:Colony Count, Microbial / Q000379:methods; D004202:Disinfectants / Q000494:pharmacology*; D004305:Dose-Response Relationship, Drug; D004950:Esterases / Q000378:metabolism; D005434:Flow Cytometry / Q000379:methods; D005456:Fluorescent Dyes / Q000378:metabolism; D005506:Food Contamination / Q000032:analysis / Q000517:prevention & control*; D005511:Food Handling / Q000379:methods*; D005516:Food Microbiology; D005519:Food Preservation / Q000379:methods; D010084:Oxidation-Reduction; D013171:Spores, Bacterial; D013696:Temperature; D013997:Time Factors
D004202:Disinfectants; D005456:Fluorescent Dyes; D000244:Adenosine Diphosphate; D000255:Adenosine Triphosphate; D004950:Esterases