Role of O-antigen on the Escherichia coli O157:H7 cells hydrophobicity, charge and ability to attach to lettuce.

Environmental factors encountered during growing and harvesting may contribute to Escherichia coli O157:H7 contamination of lettuce. Limited nutrients and extended exposure to water may cause E. coli O157:H7 to shed its O antigen. Absence of the O157-polysaccharide antigen could affect the cell's physicochemical properties (hydrophobicity and cell charge) and ultimately influence its attachment to surfaces. The objectives of this study were to evaluate the effect of the E. coli O157:H7 O-antigen on the cell's overall hydrophobicity, charge and ability to attach to cut edge and whole leaf iceberg lettuce surfaces. Three strains of E. coli O157:H7 (86-24 wild type; F-12, mutant lacking the O-antigen and pRFBE, plasmid for O157 gene reintroduced) were examined for their hydrophobicity, overall charge and ability to attach to lettuce. Overall, E. coli O157:H7 attached at higher levels to cut surfaces over whole leaf surfaces (P=0.008) for all strains and treatments. Additionally, the strain lacking the O-antigen (F12)-attached significantly less to lettuce (P=0.015) than the strains expressing the antigen (WT and pRFBE). Cells lacking the O antigen (strain F-12) were also significantly more hydrophobic than strains 86-24 or pRFBE (P≤0.05). Surface charge differed among the strains tested (P≤0.05); however, it did not appear to influence bacterial attachment to lettuce surfaces. The charge was not fully restored in the pRFBE strain (expression of O-antigen reintroduced), therefore, no conclusions can be made pertaining to the effect of charge on attachment in this study. Results indicate that E. coli O157:H7 cells which lack the O-antigen have greater hydrophobicity and attach at lower concentrations than cells expressing the O-antigen, to iceberg lettuce surfaces.

Effect of alpha-cyclodextrin-cinnamic acid inclusion complexes on populations of Escherichia coli O157:H7 and Salmonella enterica in fruit juices.

Cinnamic acid (CA), a naturally occurring organic acid found in fruits and spices, has antimicrobial activity against spoilage and pathogenic bacteria, but low aqueous solubility limits its use. The purpose of this study was to determine the effectiveness of solubility-enhancing alpha-cyclodextrin-CA inclusion complexes against Escherichia coli O157:H7 and Salmonella enterica serovars suspended in apple cider or orange juice at two different incubation temperatures (4 and 26 degrees Celsius). Two concentrations (400 and 1,000 mg/liter) of alpha-cyclodextrin-CA inclusion complex were aseptically added to apple cider inoculated with E. coli O157:H7 (7 log CFU/ml) and orange juice inoculated with a cocktail of six Salmonella enterica serovars (7 log CFU/ml). Samples were extracted at 0 min, at 2 min, and at 24-h intervals for 7 days, serially diluted in 0.1 % peptone, spread plated in duplicate onto tryptic soy agar, and incubated at 35 degrees Celsius for 24 h. Populations of E. coli O157:H7 in apple cider were significantly reduced (P < or = 0.05) during the 7-day sampling period in all solutions regardless of temperature. Compared with the controls, populations were significantly reduced by the addition of 400 and 1,000 mg/liter inclusion complex, but reductions were not significantly different (P > or = 0.05) between the two treatment groups (400 and 1,000 mg/liter). Salmonella was significantly reduced in all solutions regardless of temperature. There were significant differences between the control and each inclusion complex concentration at 4 and 26 degrees Celsius. Coupled with additional processing steps, alpha-cyclodextrin-CA inclusion complexes may provide an alternative to traditional heat processes.

Effect of acid stress, antibiotic resistance, and heat shock on the resistance of Listeria monocytogenes to UV light when suspended in distilled water and fresh brine.

Exposure to sublethal processing treatments can stimulate bacterial stress responses. The purpose of this research was to determine whether adaptation to common food processing stresses encountered during the preparation of ready-to-eat foods affects the dose of UV light required to significantly reduce Listeria monocytogenes populations in sterile distilled water and a 9% NaCl solution, using uridine as a chemical actinometer. L. monocytogenes strains N1-227 (from hot dog batter), N3-031 (from turkey franks), and R2-499 (from ready-to-eat meat) were acid stressed for 3 h at 35 degrees C in Trypticase soy broth with yeast extract acidified to pH 5.0, heat shocked for 1 h at 48 degrees C in brain heart infusion broth (BHIB), and selected for sulfanilamide resistance (512 microg/ml). These strains were then mixed in equal proportions and suspended in water and 9% NaCl solution, each containing 10(-4) M uridine. Samples were exposed to UV light (253.7 nm) for 0, 5, 10, 15, 20, 25, or 30 min. Inactivation was evaluated by surface plating onto modified Oxford agar and Trypticase soy agar with yeast extract and by enrichment in BHIB followed by incubation at 37 degrees C for 24 h. The absorbance of each sample was measured before and after irradiation to calculate the dose of UV light. There were no significant differences between population estimates based on medium or suspension solution. There were no population differences between acid-stressed and antibiotic-resistant or unstressed and heat-shocked L. monocytogenes strains. However, acid-stressed and antibiotic-resistant strains were significantly more resistant to UV light than were unstressed and heat-shocked strains (P < or = 0.05).

Internalization of Salmonella enterica serovar Montevideo into greenhouse tomato plants through contaminated irrigation water or seed stock.

Tomatoes have been linked to outbreaks of salmonellosis, demonstrating the need to identify sources of contamination. Objectives of this study included determining the ability for Salmonella enterica serovar Montevideo to be internalized into tomatoes from contaminated irrigation water and seed stock, and establishing whether Salmonella Montevideo can survive in fertilizer solutions. Six treatment groups (five plants per group) were irrigated with 350 ml of 7 log CFU/ml of Salmonella Montevideo every 14 days for 70 days, each group receiving an increased number of contaminated water events progressively: group 1 received one contaminated watering at day 0, and group 6 received a total of six contaminated waterings. Group 7 was a control, and group 8 was grown from seeds soaked in 8 log CFU/ml of Salmonella Montevideo for 24 h. All plants were watered daily with uncontaminated water. Three replications were completed. Fruit from every plant, and roots, stems, and leaves of one plant per treatment were sampled. All tomatoes were negative for Salmonella Montevideo; five root samples tested positive. For fertilizer studies, a commercially available fertilizer, two custom mixed and 1.0% dilutions of each (total of six solutions), and sterile water were inoculated with 8 log CFU/ml of Salmonella Montevideo and stored at 25 degrees C. Solutions were sampled at 24, 48, and 72 h. There were no differences (P > or = 0.05) between survival of Salmonella Montevideo in diluted fertilizers and the control. Results indicate Salmonella Montevideo is unable to contaminate tomato fruit via irrigation water and seed stock but can survive in fertilizer solutions.