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- DEVELOPING METHODS TO IDENTIFY SURROGATES FOR ESCHERICHIA COLI O157:H7 IN VALIDATION OF FRESH PRODUCE WASHING PROCESSES
- Rolfe, Catherine
- 2016, 2016-07
Cross-contamination during fresh produce washing is commonly prevented using chlorine treatment. Surrogate microorganisms have been widely...
Show moreCross-contamination during fresh produce washing is commonly prevented using chlorine treatment. Surrogate microorganisms have been widely used in process validation and to assess microbial cross-contamination. Fresh produce washing incorporates physical, chemical, biological and kinetic factors which create an intricate process for which little is known regarding surrogate selection. The purpose of this study was to identify the important elements relevant to produce washing processes and identify methods that will be used in surrogate selection. The behavior of three (3) non-pathogenic microorganisms (generic E. coli Nissle 1917 EcN, Pediococcus pentosaceus and lettuce isolate 813-F1) were examined in comparison to E. coli O157:H7 based on phenotypic similarities. Chlorine inactivation kinetics of E. coli O157:H7 and the non-pathogenic strains were evaluated with varying pH levels (6.5 and 8.0) and exposure times (3-30 seconds). Detachment of leaf-bound E. coli O157:H7 and non-pathogenic strains at different inoculation levels (approximately 2 and 6 log CFU/mL) and drying conditions (aging time, temperature) in wash water was examined. Chlorine inactivation at pH 6.5 resulted in a range of viability corresponding to E. coli O157:H7 and the non-pathogenic strains; demonstrating a sharp inactivation curve for E. coli O157:H7, EcN and P. pentosaceus. Whereas, inactivation at pH 8.0 allowed more survival relating to exposure time for all microorganisms. Detachment from inoculated leaves at 2 and 6 log CFU/mL inoculation showed steady survival levels in wash water at 0 ppm and lower survival at 1 ppm for all strains excluding 813-F1; 813-F1 was consistently less chlorine-sensitive in chlorine inactivation assays and more cross-contamination to wash water was observed for this strain. Aging time of inoculated bacteria on leaves was not seen to have remarkable effects on bacterial transfer during washing. These results suggest assay methods of chlorine inactivation at pH 6.5 and detachment with 6 log CFU/mL initial inoculation may be useful in selecting appropriate surrogates for fresh produce washing.
M.S. in Food Safety and Technology, July 2016
- Development of validation guidelines for high pressure processing to inactivate pressure resistant and matrix-adapted Escherichia coli O157:H7, Salmonella spp. and Listeria monocytogenes in treated juices
- Rolfe, Catherine
The fruit and vegetable juice industry has shown a growing trend in minimally processed juices. A frequent technology used in the functional...
Show moreThe fruit and vegetable juice industry has shown a growing trend in minimally processed juices. A frequent technology used in the functional juice division is cold pressure, which refers to the application of high pressure processing (HPP) at low temperatures for a mild treatment to inactivate foodborne pathogens instead of thermal pasteurization. HPP juice manufacturers are required to demonstrate a 5-log reduction of the pertinent microorganism to comply with FDA Juice HACCP. The effectiveness of HPP on pathogen inactivation is determinant on processing parameters, juice composition, packaging application, as well as the bacterial strains included for validation studies. Unlike thermal pasteurization, there is currently no consensus on validation study approaches for bacterial strain selection or preparation and no agreement on which HPP process parameters contribute to overall process efficacy.The purpose of this study was to develop validation guidelines for HPP inactivation and post-HPP recovery of pressure resistant and matrix-adapted Escherichia coli O157:H7, Salmonella spp., and Listeria monocytogenes in juice systems. Ten strains of each microorganism were prepared in three growth conditions (neutral, cold-adapted, or acid-adapted) and assessed for barotolerance or sensitivity. Pressure resistant and sensitive strains from each were used to evaluate HPP inactivation with increasing pressure levels (200 – 600 MPa) in two juice matrices (apple and orange). A 75-day shelf-life analysis was conducted on HPP-treated juices inoculated with acid-adapted resistant strains for each pathogen and examined for inactivation and recovery. Individual strains of E. coli O157:H7, Salmonella spp., and L. monocytogenes demonstrated significant (p <0.05) differences in reduction levels in response to pressure treatment in high acid environments. E. coli O157:H7 was the most barotolerant of the three microorganism in multiple matrices. Bacterial screening resulted in identification of pressure resistant strains E. coli O157:H7 TW14359, Salmonella Cubana, and L. monocytogenes MAD328, and pressure sensitive strains E. coli O15:H7 SEA13B88, S. Anatum, and L. monocytogenes CDC. HPP inactivation in juice matrices (apple and orange) confirmed acid adaptation as the most advantageous of the growth conditions. Shelf-life analyses reached the required 5-log reduction in HPP-treated juices immediately following pressure treatment, after 24 h in cold storage, and after 4 days of cold storage for L. monocytogenes MAD328, S. Cubana, and E. coli O157:H7 TW14359, respectively. Recovery of L. monocytogenes in orange juice was observed with prolonged cold storage time. These results suggest the preferred inoculum preparation for HPP validation studies is the use of acid-adapted, pressure resistant strains. At 586 – 600 MPa, critical inactivation (5-log reduction) was achieved during post-HPP cold storage, suggesting sufficient HPP lethality is reached at elevated pressure levels with a subsequent cold holding duration.