Cross-Laboratory Comparative Study of the Impact of Experimental and Regression Methodologies on Salmonella Thermal Inactivation Parameters in Ground Beef

ABSTRACT Isothermal inactivation studies are commonly used to quantify thermal inactivation kinetics of bacteria. Meta-analyses and comparisons utilizing results from multiple sources have revealed large variations in reported thermal resistance parameters for Salmonella, even when in similar food materials. Different laboratory or regression methodologies likely are the source of methodology-specific artifacts influencing the estimated parameters; however, such effects have not been quantified. The objective of this study was to evaluate the effects of laboratory and regression methodologies on thermal inactivation data generation, interpretation, modeling, and inherent error, based on data generated in two independent laboratories. The overall experimental design consisted of a cross-laboratory comparison using two independent laboratories (Michigan State University and U.S. Department of Agriculture, Agricultural Research Service, Eastern Regional Research Center [ERRC] laboratories), both conducting isothermal Salmonella inactivation studies (55, 60, 62°C) in ground beef, and each using two methodologies reported in prior studies. Two primary models (log-linear and Weibull) with one secondary model (Bigelow) were fitted to the resultant data using three regression methodologies (two two-step regressions and a one-step regression). Results indicated that laboratory methodology impacted the estimated D60°C- and z-values (α = 0.05), with the ERRC methodology yielding parameter estimates ~25% larger than the Michigan State University methodology, regardless of the laboratory. Regression methodology also impacted the model and parameter error estimates. Two-step regressions yielded root mean square error values on average 40% larger than the one-step regressions. The Akaike Information Criterion indicated the Weibull as the more correct model in most cases; however, caution should be used to confirm model robustness in application to real-world data. Overall, the results suggested that laboratory and regression methodologies have a large influence on resultant data and the subsequent estimation of thermal resistance parameters.

Effect of Refrigerating Delayed Shipments of Raw Ground Beef on the Detection of Salmonella Typhimurium†

In eight separate trials, four groups of raw ground beef samples were inoculated with 0.04 to 0.3 CFU/g of Salmonella Typhimurium (DT 104). Each group consisted of four 25-g samples (three inoculated and one uninoculated). After inoculation, these samples were shipped by overnight courier in shipping containers with ice packs from the U.S. Department of Agriculture (USDA), Eastern Regional Research Center, in Wyndmoor, Pa., to the U.S. Food Safety and Inspection Service (FSIS), Eastern Laboratory, in Athens, Ga. A total of 128 samples (32 in each of four groups) were shipped. A temperature data logger was placed inside each shipping container to record the temperature during shipping and storage. The first group of ground beef samples was analyzed within approximately 1 h of arrival. The second group of samples was left in the original containers, with a gel ice pack, for 24 h before processing. The third and fourth groups of samples were removed from the original shipping containers and stored at room temperature (21 ± 2°C) for 6 h and then in a refrigerator at 4 ± 2°C for 24 and 48 h, respectively, before analysis. The samples were analyzed for the presence of Salmonella according to the USDA/FSIS Microbiological Laboratory Guidebook, chapter 4.02. There was no significant difference in the presence and levels of Salmonella in ground beef among the four test groups. These data show that it is acceptable to process the late-arriving ground beef samples for the detection of Salmonella if they are kept in a refrigerator (4 ± 2°C) for 24 to 48 h or when the shipments arrive late (24 h in the container with ice pack).

Recovery Rate of Listeria monocytogenes from Commercially Prepared Frankfurters during Extended Refrigerated Storage†

To assess the prevalence of Listeria monocytogenes in vacuum-sealed packages of frankfurters, about 33,000 packages (1 lb each) were obtained by a third-party contractor from 12 volunteer commercial manufacturers over a 2-year period. The 12 producers, each of which contributed about 2,700 packages of frankfurters from one production run, comprised 9 large and 3 small plants located in eight U.S. Department of Agriculture/Food Safety and Inspection Service (USDA/FSIS) districts in 10 states. Five days after manufacture, 500 packages were sampled at the USDA/Agricultural Research Service (ARS) Eastern Regional Research Center (ERRC) in Wyndmoor, Pa., by the USDA/ARS package rinse method. At regular intervals during subsequent storage at 4 and 10°C, an additional 200 packages were tested for the pathogen at each sampling point. From a statistical perspective, L. monocytogenes was not recovered from any of the products of nine of the producers, whereas the pathogen was recovered at rates of 1.5% (plant 367), 2.2% (plant 439), and 16% (plant 133) from the products of the remaining three plants. In total, 532 of 32,800 (1.6%) packages of frankfurters tested positive for the pathogen. The recovery rates did not change appreciably over time, there was no appreciable difference in L. monocytogenes recovery rates with respect to frankfurter storage temperature (4 or 10°C), and the seasonality of manufacture had no influence on recovery rate. Molecular subtyping of multiple L. monocytogenes–positive isolates from each plant revealed that profile A (serotype 1/2a) was displayed by about 90% of the 1,105 isolates tested. However, in some cases it was also possible to recover more than one profile from a given plant. This study provides estimates of the prevalence, types, and viability of L. monocytogenes associated with commercially prepared frankfurters during extended refrigerated storage.

Listeria Methods Development Research at the Eastern Regional Research Center, U.S. Department of Agriculture

Listeria methods research at the U.S. Department of Agriculture, Eastern Regional Research Center, has concentrated on 2 areas during the past year. The first was development of techniques for assessing isolation methods for their ability to detect sublethally stressed cells. It appears that a number of widely used media do not accurately detect Listeria that have been injured by thermal processing or acidification. The second was development of improved plating media. One, modified Vogel-Johnson agar, shows promise; it is highly selective and quantitative, and eliminates the need to select colonies on the basis of a blue color when illuminated with reflected light

Wiley Led the Way: A Century of Federal Honey Research

Jonathan W. White, Jr, received the AOAC Harvey W. Wiley Award at the 100th AOAC Annual International Meeting and Exposition in Scottsdale, AZ, September 15, 1986. In 1942, after earning his Ph.D. at Purdue University, White became assistant chemist at the Eastern Regional Research Center (ERRC) of the U.S. Department of Agriculture at Wyndmoor, PA, where he spent almost his entire professional career. In 1948, he began the extensive research on honey for which he was selected as the 1986 Wiley Award recipient. He was named chief of the ERRC Plant Products Laboratory in 1965; 10 years later he requested reassignment to conduct research on the detection of adulteration of honey by high-fructose corn syrup. He retired from USDA in 1978 and until recently headed a honey testing service called Honeytech, Inc. White was elected a Fellow of AOAC in 1969 and was named Associate Referee of the Year in 1977

Comparison of Three Methods for Determination of N-NitrosopyrroIidine in Fried Dry-Cured Bacon

The recently developed Eastern Regional Research Center (ERRC) dry column chromatographic procedure for determining A'-nitrosopyrrolidine (NPYR) in fried cure-pumped bacon was evaluated for its applicability to fried dry-cured bacon. The method was then compared with 2 established procedures for volatile nitrosamine analysis in cured meat products: the multidetection thermal energy analyzer (MD) method and the mineral oil distillation (MOD) screening procedure. No significant difference (P < 0.05) in NPYR values was found between the ERRC and MD procedures, but significant differences were found between the ERRC and MOD procedures and between the MOD and MD procedures. No artifactual nitrosamine formation was found in the ERRC procedure, but significant amounts were found in samples analyzed by the MOD procedure. The ERRC method was demonstrated to be rugged and very rapid. It is proposed that the ERRC method replace the MOD method as the official screening procedure for NPYR in fried bacon.