Salmonella isolated from ready-to-eat pasteurized liquid egg products: Thermal resistance, biochemical profile, and fatty acid analysis.

The Egg Products Inspection Act of 1970 requires that egg products in the U.S. must be pasteurized prior to release into commerce. The USDA Food Safety and Inspection Service (FSIS) is responsible for regulating egg products. Salmonellae are infrequently isolated from pasteurized egg products by food manufacturers or the FSIS and may be present as a result of either pasteurization-resistant bacteria or post-processing contamination. In this study, seventeen strains of Salmonella isolated from pasteurized egg products and three heat-resistant control strains were compared for the following attributes: thermal resistance in liquid whole egg (LWE) at 60 °C, enzymatic profiles, and serotyping and phage typing, antibiotic susceptibility, fatty acid analysis and strain morphological variation evaluated by scanning electron microscopy. Isolates were serotyped as Heidelberg (4 isolates), Widemarsh, Mbandaka, Cerro, Thompson, 4,12:i:-, and Enteritidis (8 isolates). All 20 isolates were sensitive to all 14 antibiotics tested for. The D60 values in LWE ranged from 0.34 to 0.58 min. All 20 strains were recovered from LWE inoculated with 8.5 logCFU/mL of Salmonella and pasteurized at 60 °C for 3.5 min; however, some isolates were not recovered from pasteurized LWE that had been inoculated with only 4.5 logCFU/mL Salmonella and treated at 60 °C for 3.5 min. Although some strains exhibited atypical enzymatic activity (e.g., reduction of adonitol, hydrolysis of proline nitroanilide or p-n-p-beta-glucuronide, and nonreduction of melibiose), differences in biochemical reactions could not be correlated with differences in thermal resistance. Furthermore, fatty acid analysis revealed that differences insaturate/unsaturated profiles may be correlated with differences in heat resistance, in two instances. One heat resistant strain (#13, Enteritidis) had the statistically lowest unsaturated/saturate ratio at 39%. However, one heat sensitive strain (#3, serovar 4,12:i:-) had the highest unsaturated/saturate ratio at 81%, and also the lowest concentration of stearic acid. This data represents the first steps in determining whether Salmonella contamination in pasteurized egg products may be the result of either thermally-resistant isolates or post-processing contamination. Contamination of LWE by Salmonella strains with higher heat resistance, (e.g., isolate #'s 2, 6, 10 and 12) may indicate the ability of Salmonella to survive pasteurization, while contamination of LWE strains with lower heat resistance (e.g., isolate #'s 1, 3, 5, 7, 8, 11, and 15) may indicate post-processing contamination of LWE by this foodborne pathogen.

Influence of mycorrhizal fungi on fate of E. coli O157:H7 and Salmonella in soil and internalization into Romaine lettuce plants.

The objectives of this study were to determine the influence of a symbiotic arbuscular mycorrhizal (AM) fungus on persistence of Salmonella and enterohemorrhagic Escherichia coli O157:H7 (EHEC) within soil, and survival within Romaine lettuce. Romaine seedlings were grown with or without AM fungi. Soil surrounding plants was inoculated with ca. 8 log CFU/plant of either Salmonella enterica or E. coli EHEC composites. Samples (soil, root, and shoot) were analyzed on days 1, 8, 15 and 22 for Salmonella and EHEC by direct plating and selective enrichment. Twenty-four hours after inoculation, populations of Salmonella and EHEC, respectively, were 4.20 and 3.24 log CFU/root, 2.52 and 1.17 log CFU/shoot, and 5.46 and 5.17 log CFU/g soil. By selective enrichment, samples tested positive for Salmonella or EHEC at day 22 at rates of 94 and 68% (shoot), 97 and 56% (root), and 100 and 75% (soil), respectively, suggesting that Salmonella has a greater propensity for survival than EHEC. Salmonella populations in soil remained as high as 4.35 log CFU/g by day 22, while EHEC populations dropped to 1.12 log CFU/g in the same amount of time. Ninety-two percent of all Romaine leaves in our study were positive for internalized Salmonella from days 8 to 22 and remained as high as 1.26 log CFU/shoot on day 22 in AM fungi+Romaine plants. There were no differences (P>0.05) between the survival of either pathogen based on the presence or absence of mycorrhizal fungi. Results of this study suggest that AM fungi do not affect the internalization and/or survival of either S. enterica or E. coli O157:H7 in Romaine lettuce seedlings. Our results should provide Romaine lettuce farmers confidence that the presence and/or application of AM fungi to crop soil is not a contributing factor to the internalization and survival of Salmonella or E. coli O157:H7 within Romaine lettuce plants.

Reduction of an E. coli O157:H7 and Salmonella composite on fresh strawberries by varying antimicrobial washes and vacuum perfusion.

A 2011 outbreak of hemorrhagic colitis, which resulted in the death of two individuals, was associated with contaminated strawberries. A study was conducted to identify antimicrobial washes effective at reducing E. coli O157:H7 and Salmonella enterica from the surface of fresh whole strawberries during two-minute immersion washes. Twenty-seven antimicrobial treatments were tested. Vacuum perfusion was applied to strawberries during chlorine and peracetic acid treatments to promote infiltration of sanitizer into porous strawberry tissue. Strawberries were inoculated to 7.1logCFU/strawberry with a seven-strain bacterial composite, consisting of three strains of E. coli O157:H7 and four serovars of Salmonella enterica. Berries were air-dried for 2h and immersed in circulating antimicrobial solutions for 120s at 22°C. Four treatments reduced ≥3.0logCFU/strawberry, including (a) 1% acetic acid+1% H2O2, (b) 30% ethanol+1% H2O2, (c) 90ppm peracetic acid, and (d) 1% lactic acid+1% H2O2. Two additional treatments that reduced 2.8logCFU/strawberry were (a) 40% ethanol, and (b) 1% each of phosphoric+fumaric acids. Eight treatments reduced 2.0-2.6logCFU/strawberry. Five treatments reduced <1.45CFU/strawberry, including (a) 1% citric acid, (b) 1% lactic acid, (c) 1% acetic acid, (d) 0.5% each of acetic+citric acids and (e) 0.5% each of acetic+lactic acids. The use of vacuum perfusion with 200ppm chlorine or 90ppm peracetic acid did not reduce greater populations of pathogens than did the same treatments without vacuum perfusion. Fourteen treatments reduced no more pathogens (p<0.05) than did sterile deionized water. Results from this study provide some options for end-point decontamination of strawberries for retail operations just prior to serving to customers.

Kinetics model comparison for the inactivation of Salmonella serotypes Enteritidis and Oranienburg in 10% salted liquid whole egg.

The goal of this study was to determine the inactivation kinetics of Salmonella in commercial 10% salted liquid whole egg (LWE) to assist the U.S. Department of Agriculture in writing new liquid egg pasteurization guidelines. Current data are not sufficient for predicting thermal inactivation kinetics of Salmonella spp. for use in updating pasteurization guidelines for many types of liquid egg products, including salted LWE (SLWE). This is, in part, due to variations in Salmonella strains and changes in the processing of liquid egg products that have arisen in the past 40 years. Pasteurization guidelines are currently being reevaluated in light of recent risk assessments. Heat-resistant Salmonella serovars Enteritidis and Oranienburg were composited and mixed into 10% SLWE, resulting in final populations of approximately 5.7-7.8 log colony-forming units (CFU)/mL. Inoculated egg was injected into glass capillary tubes, flame-sealed, and heated in a water bath at 60, 62.2, 63.3, 64.3, or 66°C. Contents were surface-plated and incubated at 37°C for 24 h. Survival curves were not log-linear (log levels versus time), but decreased rapidly, and after initial periods became linear. Asymptotic decimal reduction values at each temperature were calculated from survivor curves with a minimum inactivation of 5.0 log CFU/mL. The asymptotic thermal D-values for SLWE were 3.47, 2.23, 1.79, 1.46, and 1.04 min at 60, 62.2, 63.3, 64.3, or 66°C, respectively. The calculated thermal z-value was 11.5°C. A model that predicts lethality for given times and temperatures that was developed predicted that the current pasteurization requirements for 10% SLWE (i.e., 63.3°C for 3.5 min, or 62.2°C for 6.2 min) are not sufficient to inactivate 7 log CFU/mL of Salmonella and only achieve approximately 4 log CFU/mL inactivation. This model will assist egg-products manufacturers and regulatory agencies in designing pasteurization processes to ensure product safety.