No description available.

Listeria monocytogenes is a major concern for the food industry. It is one of the major agents causing listeriosis. The objective of the first part of this study is to evaluate the effect of hydration level on attached listeria monocytogenes on stainless steel/High density polyethylene transferred to food products. Attached cells were prepared on stainless steel/High density polyethylene. Transfer experiments were conducted from inoculated surface material slides to cold smoked salmon fillets. This experiment was repeated 6 times. The results were analyzed with an analysis of variance by SAS. The differences between the different RH% and surface conditions were not statistically significant. There was variability in between packages, brands and over the course of storage after opening, and likely contributed to the variability of transfer observed in this set of experiments. The objective of the second part of the research is to study the effect of hydration level on the detachment of Listeria monocytogenes biofilm growing on stainless steel by using Atomic force microscope. Biofilms were grown on stainless steel in drip flow bioreactor at 32 °C for 72 h. Then biofilms were equilibrated over saturated salt solution at 20 °C for 48 h before the Atomic Force Microscope experiment. The results showed that cohesive energy value of the biofilm increased with biofilm depth. Only square shaped displaced 2.5X2.5 μm region were visualized after serious of raster scanning under high load which means that moisture condition of Listeria monocytogenes biofilm can significantly affect the cohesiveness between of Listeria monocytogenes biofilm.

The food-borne pathogen Listeria monocytogenes can attach to the environmental surfaces and develop biofilm which can cause food contamination in the food industries. Sortase A and surface proteins are involved in biofilm and virulence of L. monocytogenes. Curcumin was reported to inhibit sortase A and biofilm in gram positive bacteria. The overall objective of this study was to observe the effect of curcumin (Curcuma longa) on the biofilm formation and surface proteins of L. monocytogenes. The antibiofilm effect of curcumin against the strain LM21 (wild type) and s22-11G (sortase A defective mutant) was studied using the microtiter plate assay. No significant differences between the growth of the wild type and the sortase A defective mutant were observed at sub-inhibitory concentrations of curcumin. However, a greater biofilm reduction was observed in the strain s22-11G. The effect of curcumin from two different manufacturers on the wild type was also compared by the microtiter plate assay. Both curcumin did not exhibit statistically different effect on the growth of the wild type. However, a greater biofilm inhibitory effect was observed in one curcumin. The HPLC results suggested that curcumin with the greater antibiofilm activity contained higher amount of curcumin which was reported to be the most potent curcuminoid compound in curcumin. Three different protein extraction methods were evaluated and the most efficient method was used for 2D-GE. When cells were grown in the presence of curcumin, 5 proteins, 16 proteins and 4 proteins were up-regulated, down-regulated and absent, respectively in L. monocytogenes LM21. The influence of the enzyme sortase A upon surface protein expression was evaluated by comparing proteins expressed by wildtype L. monocytogenes LM21 to that of the sortase A mutant, s22-11G. In strain s22-11G, 2 proteins, 8 proteins and 3 proteins were up-regulated, down-regulated and absent in comparison to wildype LM21. The exact information of these differentially expressed proteins still need to be identified by mass spectrometry.

Listeria monocytogenes is an important foodborne pathogen that can adapt to stress conditions to persist in food processing environments. Our findings show that there was a development of low-level tolerance to quaternary ammonium compound (QAC) and antibiotics ciprofloxacin and trimethoprim in L. monocytogenes after sublethal adaptation to QAC. Using eight L. monocytogenes strains, we determined the changes in MIC, growth rate, and surviving CFU for homologous and heterologous stress-response after sublethal exposure to daily cycles of fixed or gradually increasing concentration of QAC. Three main findings were observed: (1) Short-range MIC of QAC, ciprofloxacin, and trimethoprim increased by 1.6 to 2.3, 1.5 to 2.9, and 1.7 to 2.5 fold against QAC-adapted phenotypes of L. monocytogenes as compared to the non-adapted cells; (2) QAC-adapted phenotypes of L. monocytogenes exhibited a significant increase in growth rate by 2.5 to 7.1, 2.1 to 6.8, or 1.4 to 4.8 fold in the broth model containing QAC, ciprofloxacin, or trimethoprim respectively, as compared to non-adapted cells; and (3) QAC-adapted phenotypes of L. monocytogenes exhibited a significant increase in survival by 1.5 to 4, 2.2 to 4.3, or 1.3 to 3.2 log CFU/ml in the agar model containing QAC, ciprofloxacin, or trimethoprim respectively, as compared to non-adapted cells (P < 0.05). There were strain differences in QAC-adapted phenotypes of L. monocytogenes for both homologous and heterologous stress-response with some strains exhibiting a significant increase in short-range MIC, growth rate, and survival while others exhibiting no changes as compared to non-adapted cells. These findings suggest the potential formation of low-level QAC-tolerant and antibiotic-tolerant phenotypes in some L. monocytogenes strains under residual QAC concentrations (where QAC may be used widely) and such cells if not inactivated may survive longer to increase food safety risk.

The fresh-cut produce industry has seen expansive growth in recent years, to meet consumer demand ready-to-eat (RTE) salads have included the use of non-traditional ingredients. Uncommon ingredients include beet greens, kale, broccoli stalk, and Brussels sprouts, since these ingredients have not historically been consumed raw, potential food safety issues should be reassessed. Current processing technologies include produce washes that can reduce microbial levels but do not eradicate all populations. The lack of a kill step in produce processing emphasizes the need to minimize pathogen contamination during production and growth during a product’s shelf life. Listeria monocytogenes, a leading cause of foodborne illness related deaths, continues to challenge the industry with recent outbreaks and recalls of fresh-cut produce. These events present the need to better understand L. monocytogenes growth potential in RTE produce during storage and distribution. Traditional salad greens have been researched extensively, however, limited knowledge is available on new inclusions. While temperature is known to strongly influence microbial growth, the effects of physical abuse during storage and distribution are unknown. The purpose of this study was to characterize L. innocua’s, a surrogate for L. monocytogenes, growth behavior in processed beet greens, kale, broccoli stalk, and Brussels sprouts when exposed to simulated physical and thermal abuses during storage and distribution. To evaluate L. innocua growth during storage and distribution produce samples were obtained from a local processor in retail packaging and surface inoculated. The samples were conditioned at 4℃ for 18h prior to being exposed to a series of physical abuses (compression, drop, and vibration) typical of storage and distribution. After abuse, produce was incubated at 4 or 8°C and sampled post-abuse through 16 and 11 days, respectively. Samples were enumerated for L. innocua, aerobic and psychrotrophic microorganisms, and lactic acid bacteria. To monitor growing conditions in each vegetable, product pH, water activity, and headspace (gas analysis), were observed at each time pull. The study found physical abuse had no significant effect on L. innocua, or microbiota growth regardless of vegetable or incubation temperature (P > 0.05). Vegetable intrinsic factors (pH, Aw, and headspace) did not seem to interfere in L. innocua or background microbiota growth during incubation. All vegetables supported L. innocua growth under 8℃. Growth of L. innocua was greatest in beet greens, followed by kale, broccoli stalk, and Brussels sprouts in descending order. Significant growth of L. innocua at 4 and 8ᵒC was seen on day 6 and 4 in beet greens, 11 and 6 in Brussels sprouts, 16 and 4 in kale, and 16 and 6 in broccoli stalk (P < 0.05). Overall, these results show the studied RTE vegetables can support L. monocytogenes growth during storage and distribution, especially under abusive temperatures, demonstrating the importance of prevention strategies during processing and refrigeration throughout RTE produce shelf life.

No description available.

No description available.

The heat resistance was determined for Salmonella typhimurium and Listeria monocytogenes Scott A suspended in 2% UHT processed milk and in a biofilm matrix. Pure cultures at an initial concentration of 10⁵ / ml were used. Heat resistance was determined by two methods. One method was sealed borosilicate glass TDT tubes that were completely submerged in the heating menstrum. Biofilms were grown on Buna-n rubber o-rings (4.46 mm O.D. x 1.41 mm]. D.) for 36 hours. All other cultures used were in stationary phase of growth. The three treatments tested were: inoculated milk, sterile milk and a biofilm on an o-ring, and inoculated milk with a sterile o-ring. At the three temperatures tested (60, 63, 67°C), there was no significant difference (p>0.05) in D-values between treatments. There was a significant difference (p<0.001) between the D-values for Salmonella and Listeria. The second method used a laboratory scale HTST pasteurizer to determine the difference in heat resistance of the same organisms suspended in 2% milk vs. sloughed off pieces of biofilm in milk. Pure cultures of the organisms at an initial inoculum of 10⁵ / ml were used. Flow rates of the pasteurizer were adjusted to achieve two different F-values for each organism at a reference temperature of 71.7°C. Neither S. typhimurium nor L. monocytogenes Scott A was recovered from pasteurized samples of either treatment. The heating involved in come up and cool down of the transit lines was considered in determining F-value. Under commercial HTST processing, concentrations of 10⁵ / ml of S. typhimurium and L. monocytogenes Scott A would not survive pasteurization. The results also show that if pieces of biofilms (3.8 x 10⁻⁴ mm²- 8.8 x 10⁻³ mm²) were sloughed off gaskets in the processing lines they would not survive pasteurization. The heating characteristics of these two systems were so dissimilar they could not be compared. It should however be noted that in the TDT tubes it was necessary to obtain a slightly higher F-value before no growth was seen as compared with the pasteurizer. In the pasteurizer the laminar flow properties would contribute to a more uniform heating. The TDT tube experiences convection heating which can produce cold spots in the tubes and could explain the need for an increased F-value. / Master of Science

Ready-to-eat meat products have been implicated in several foodborne listeriosis outbreaks. Microbial contamination of these products can occur after the product has been thermally processed and is being rapidly chilled using salt brines. The objective of this study was to determine the effect of ultraviolet irradiation on the inactivation of Listeria monocytogenes and lactic acid bacteria in a model brine chiller system. Two concentrations of brines (7.9%w/w or 13.2%w/w) were inoculated with a ~6.0 log10 CFU/ml cocktail of L. monocytogenes or lactic acid bacteria and passed through the ultraviolet (UV) treatment system for 60 minutes. Three replications of each bacteria and brine combination were performed and resulted in at least a 4.5 log reduction in microbial numbers in all treated brines after exposure to ultraviolet light. Bacterial populations were significantly reduced after five minutes exposure to UV light in the model brine chiller as compared to the control, which received no UV light exposure (P<0.05). The maximum rate of inactivation for both microorganisms in treated brines occurred between minute 1 and 15 of ultraviolet exposure. Overall, results indicate that inline treatment of chill brines with ultraviolet light (UVC) shows promise in inactivating L. monocytogenes and lactic acid bacteria. Due to the low capital involved in initiating a continuous inline UV system, the use of ultraviolet energy may prove to be beneficial for effectively controlling pathogens in recycled chill brines without interrupting the chilling operation. An inline ultraviolet system could be used in a hazard analysis and critical control points plan. / Master of Science

Since 2006 there have been four Salmonella enterica and one Listeria monocytogenes foodborne outbreaks linked to whole cantaloupe fruit. No post-harvest intervention to reduce potential contamination on cantaloupe currently exists. The complex surface topography of netted cantaloupes aids bacterial attachment. This research evaluates the use of allyl isothiocyanate (AITC; a natural antimicrobial) to reduce populations of S. enterica Michigan and L. monocytogenes on the surface of cantaloupe. Fifty μl of S. Michigan or L. monocytogenes was inoculated onto whole ‗Athena‘ or ‗Hales Best Jumbo‘ (‗HBJ‘) cantaloupe fruit in 22 mm diameter circles and allowed to dry for 90 min. resulting in 6.60 log CFU/g. Cantaloupe received either AITC liquid or vapor, sterile deionized water, 200 ppm sodium hypochlorite per circle, or no treatment. All cantaloupes were stored in separate sealed glass desiccators for 1 or 24 h at 25°C or 35°C. To enumerate the bacteria following treatment, 22 mm sections of the rind were removed, homogenized and plated onto appropriate agar. Headspace analysis using Gas Chromatography-Mass Spectrometry (GC-MS) quantified the concentration of each AITC vapor treatment. The texture quality of the pericarp tissue of whole cantaloupes was evaluated after 24 h treatments, followed by two weeks of storage at 4°C. The concentration of vapor ranged from 3.4 to 19.6 μl AITC/L inside the desiccators. The liquid treatment reduced (P < 0.05) S. Michigan populations on ‗Athena‘ (3 log CFU/g) and L. monocytogenes on ‗HBJ‘ (2.6 log CFU/g). The longer exposure time to the AITC vapor (24 h versus 1 h) resulted in a greater reduction of both S. Michigan and L. monocytogenes on ‗Athena‘ and treatments at 35°C reduced microbial populations up to 4.5 times greater (P < 0.05). The highest vapor concentration reduced (P < 0.05) both pathogens at least 3.0 log CFU/g on ‗Athena‘ at 25°C. Generally, bacterial pathogens from the surface of ‗Athena‘ cantaloupe were reduced more than pathogens inoculated on the surface of ‗HBJ.‘ The application of AITC liquid or vapor is a natural alternative post-harvest treatment to 200 ppm free chlorine to reduce the level of bacterial contamination on cantaloupe surfaces for certified organic production. / Ph. D.