Sensitivity and specificity of the BAX for screening/Listeria monocytogenes assay: internal validation and independent laboratory study.

In this study to certify the BAX for Screening/Listeria monocytogenes assay (DuPont Qualicon, Wilmington, DE), an internal evaluation was conducted on 16 food types that were simultaneously analyzed with the BAX system (BAX), and the ISO method for the detection of L. monocytogenes (ISO). No statistically significant difference in performance between the BAX and ISO methods was observed. Inclusivity/exclusivity testing showed that the BAX system was able to detect 97 of 97 (100%) of L. monocytogenes strains tested. None of 56 other Listeria species or non-Listeria tested gave a reproducible positive BAX result. Ruggedness testing demonstrated that performance of the assay was not affected by reasonable variability in the operating parameters. BAX was then submitted for independent laboratory validation. In this phase, BAX was compared with standard culture methods for the detection of L. monocytogenes in chicken (USDA-FSIS), crab meat (BAM), and milk (AOAC). This study validated product claims of sensitivity and specificity >98% in accordance with AOAC Performance Tested Method requirements.

Comparative survival of Salmonella typhimurium DT 104, Listeria monocytogenes, and Escherichia coli O157:H7 in preservative-free apple cider and simulated gastric fluid.

This study compared the survival of three-strain mixtures (ca. 10(7) CFU ml(-1) each) of Salmonella typhimurium DT104, Listeria monocytogenes, and Escherichia coli O157:H7 in pasteurized and unpasteurized preservative-free apple cider (pH 3.3-3.5) during storage at 4 and 10 degrees C for up to 21 days. S. typhimurium DT104 populations decreased by <4.5 log10 CFU ml(-1) during 14 days storage at 4 and 10 degrees C in pasteurized cider, and by > or =5.5 log10 CFU ml(-1) during 14 days in unpasteurized cider stored at these temperatures. However, after 7 days at 4 degrees C, the S. typhimurium DT104 populations had decreased by only about 2.5 log10 CFU ml(-1) in both pasteurized and unpasteurized cider. Listeria monocytogenes populations decreased below the plating detection limit (10 CFU ml(-1)) within 2 days under all conditions tested. Survival of E. coli O157:H7 was similar to that of S. typhimurium DT104 in pasteurized cider at both 4 and 10 degrees C over the 21-days storage period, but E. coli O157:H7 survived better (ca. 5.0 log10 CFU ml(-1) decrease) than S. typhimurium DT104 (> 7.0 log10 CFU ml(-1) decrease) after 14 days at 4 degrees C in unpasteurized cider. In related experiments, when incubated in simulated gastric fluid (pH 1.5) at 37 degrees C, S. typhimurium DT104 and L. monocytogenes were eliminated (5.5-6.0 log10 CFU ml(-1) decrease) within 5 and 30 min, respectively, whereas E. coli O157:H7 concentrations decreased only 1.60-2.80 log10 CFU ml(-1) within 2 h.

Survival of Escherichia coli O157:H7 in full- and reduced-fat pepperoni after manufacture of sticks, storage of slices at 4 degrees C or 21 degrees C under air and vacuum, and baking of slices on frozen pizza at 135, 191 and 246 degrees C.

Pepperoni batter was prepared with fat contents of about 15, 20, and 32% (wt/wt) and inoculated with a pediococcal starter culture and > or = 2.0 x 10(7) CFU/g of a five-strain inoculum of Escherichia coli O157:H7. The batter was fermented at 96 degrees F (ca. 36 degrees C and 85% relative humidity (RH) to pH < or = 4.8 and then dried at 55 degrees F (ca. 13 degrees C) and 65% RH to a moisture/protein ratio of < or = 1.6:1. For storage, slices were packaged under air or vacuum and stored at 39 degrees F (ca. 4 degrees C) and 70 degrees F (ca. 21 degrees C). For baking, frozen slices were placed on retail frozen cheese pizzas that were subsequently baked at 275 degrees F (ca. 135 degrees C), 375 degrees F (ca. 191 degrees C), or 475 degrees F (ca. 246 degrees C) for 0 to 20 min. Appreciable differences related to fat levels were observed after drying; pathogen numbers decreased by 1.04, 1.31 and 1.62 log10 units in sticks prepared from batter at initial fat levels of 15, 20, and 32%, respectively. During storage, the temperature rather than the atmosphere had the greater effect on pathogen numbers, with similar viability observed among the three fat levels tested. At 70 degrees F (ca. 21 degrees C), compared to original levels, pathogen numbers decreased by > or = 5.56 and > or = 4.53 log10 units within 14 days in slices stored under air and vacuum, respectively, whereas at 39 degrees F (ca. 4 degrees C) numbers decreased by < or = 2.43 log10 CFU/g after 60 days of storage under either atmosphere. Baking, as expected, resulted in greater reductions in pathogen numbers as the temperature and/or time of baking increased. However, it was still possible to recover the pathogen by enrichment after baking frozen slices on frozen pizza at 475 degrees F (ca. 246 degrees C) for 10 min or at 375 degrees F (ca. 191 degrees C) for 15 min. The calculated D values for all three temperatures tested increased as the fat content of the batter increased from 15 to 20 to 32%. The present study confirmed that fermentation and drying were sufficient to reduce levels of E. coli O157:H7 in pepperoni sticks by < 2.0 log10 CFU/g. Storage of slices for at least 14 days at ambient temperature under air resulted in a > 5.5-log10-unit total reduction of the pathogen. Baking slices on frozen pizza for at least 15 min at 475 degrees F (ca. 246 degrees C) or 20 min at 375 degrees F (ca. 191 degrees C) was necessary to reduce pathogen numbers to below detection by both direct plating and enrichment.

Behavior of Salmonella typhimurium DT104 during the manufacture and storage of pepperoni.

Pepperoni batter (ca. 70% pork:30% beef) was prepared and subsequently inoculated with a six-strain cocktail (ca. 4.4 x 10(7) per gram batter) of Salmonella typhimurium DT104. After fermentation at 36 degrees C and 92% relative humidity (RH) to < or = pH 4.8, counts of the pathogen decreased by about 1.3 log10 units. An additional 1.6 log10 unit decrease was observed following drying at 13 degrees C and 65% RH to a moisture protein ratio (M/Pr) of 1.6:1. After storage of pepperoni sticks for 56 days under vacuum at 4 or 21 degrees C, counts of the pathogen were about 4.6 and 6.6 log10 units lower, respectively, compared with starting levels in the batter. These data establish that fermentation and drying result in about a 3.0 log10 reduction in numbers of S typhimurium DT104 in pepperoni sticks and that storage of pepperoni sticks under vacuum at ambient temperature is more severe on the pathogen than refrigerated storage.