Short communication: Decimal log reductions of Salmonella Senftenberg 775 W and other Salmonella serovars in nonfat milk and powder.

This study aimed to compare the thermal resistance of Salmonella Senftenberg 775 W with other serovars of Salmonella in nonfat dry milk (NDM) and hydrated NDM. The scientific literature suggests that Salmonella Senftenberg 775 W is the most heat-resistant serovar in high-water-activity foods such as milk, but little is known about the heat resistance of Salmonella Senftenberg 775 W compared with other Salmonella serovars in low-water-activity foods such as NDM. The 5 serovars of Salmonella used in this study were Enteritidis, Montevideo, Newport, Senftenberg, and Typhimurium. The hydration of NDM was conducted at 13% (wt/vol) total solids. The NDM was inoculated with the 5 individual serovars of Salmonella and dried again to its original pre-inoculation water activity. Hydrated NDM was prepared from individually inoculated NDM. The surviving Salmonella population at predetermined time-temperature intervals were enumerated using injury-recovery medium, and the average log reductions for the individual serovars were calculated. As expected, Salmonella Senftenberg 775 W was the most heat-resistant serovar in hydrated NDM at 59°C and 65°C. However, the heat resistance of Salmonella Senftenberg 775 W was found to be lower than or comparable to that of other serovars in low-water-activity NDM at 80°C and 90°C.

Hyperspectral imaging of foodborne pathogens at colony and cellular levels for rapid identification in dairy products.

This study evaluated the efficacy of hyperspectral imaging (HSI) for the rapid identification of pathogens in dairy products at the colony and cellular levels. The colony and cellular levels studies were designed as completely randomized with six replications. Three strains of Listeria monocytogenes, four strains of Escherichia coli O157: H7, Big Six Shiga toxin-producing E. coli, three strains of Staphylococcus aureus, and ten serovars of Salmonella were used in this study. Pure cultures were streaked for isolation on respective selective media, and hyperspectral data (400-1100 nm wavelength) at the colony and cellular levels were collected and stored as reference libraries. Whole milk and whole milk powder were artificially inoculated (<10 CFU/g or mL) with individual pathogenic strains/serovars. All milk and milk powder samples were enriched using brain heart infusion (BHI) broth at 37°C for 24 h, streaked for isolation on the respective selective media, and hyperspectral data for individual pathogenic strains/serovars at the colony and cellular levels were acquired and treated as test samples data. The acquired colony or cellular images were imported into ENVI software and three regions of interest were selected for each image to obtain hyperspectral data for reference libraries and test samples. Using the kNN classifier and cross-validation technique, overall classification accuracies of 90.38% and 34% were obtained for the colony- and cellular-level identification, respectively. The individual classification accuracies of pathogens in dairy products at the colony level varied between 77.5% to 100%, whereas the accuracy varied between 2.78% and 49.17% for the cellular level.

Developing an affordable hyperspectral imaging system for rapid identification of Escherichia coli O157:H7 and Listeria monocytogenes in dairy products.

The objective of this foundational study was to develop and evaluate the efficacy of an affordable hyperspectral imaging (HSI) system to identify single and mixed strains of foodborne pathogens in dairy products. This study was designed as a completely randomized design with three replications. Three strains each of Escherichia coli O157:H7 and Listeria monocytogenes were evaluated either as single or mixed strains with the HSI system in growth media and selected dairy products (whole milk, and cottage and cheddar cheeses). Test samples from freshly prepared single or mixed strains of pathogens in growth media or inoculated dairy products were streaked onto selective media (PALCAM and/or Sorbitol MacConkey agar) for isolation. An isolated colony was selected and mixed with 1 ml of HPLC grade water, vortexed for 1 min, and spread over a microscope slide. Images were captured at 2000× magnification on the built HSI system at wavelengths ranging from 400 nm to 1100 nm with 5-nm band intervals. For each image, three cells were selected as regions of interest (ROIs) to obtain hyperspectral signatures of respective bacteria. Reference pathogen libraries were created using growth media, and then test pathogenic cells were classified by their hyperspectral signatures as either L. monocytogenes or E. coli O157:H7 using k-nearest neighbor (kNN) and cross-validation technique in R-software. With the implementation of kNN (k = 3), overall classification accuracies of 58.97% and 61.53% were obtained for E. coli O157:H7 and L. monocytogenes, respectively.

Heat resistance comparison of Salmonella and Enterococcus faecium in cornmeal at different moisture levels.

Adequate surrogate identification is critical for validating in-plant thermal process controls for Salmonella inactivation in different food matrices. This study compared the thermal inactivation parameters (D- and z-values) and evaluated the heat resistance of Enterococcus faecium (8459) as a surrogate for a 5-serovar Salmonella cocktail in cornmeal. The cornmeal was spray inoculated with the respective bacteria to achieve ~9 log CFU/g population and set to the desired moisture contents (16, 22, and 28% w.b.). The inoculated cornmeal was then heat-treated at pre-determined temperatures (60, 64, and 68 °C) in sealed aluminum thermal-death-time disks in hot water baths for pre-determined time intervals. Injury-recovery media [brain heart infusion (BHI) agar overlaid with xylose lysine deoxycholate (XLD) agar for Salmonella or BHI agar overlaid m-enterococcus agar for E. faecium] were used for microbial enumeration to account for thermally injured bacterial cells. The D-values of Salmonella in cornmeal at 16, 22, and 28% moisture content were 37.5, 8.4, and 2.4 min at 60 °C, 19.9, 3.5, and 1.1 min at 64 °C, and 10.1, 1.4, and 0.5 min at 68 °C, respectively. The D-values of E. faecium in cornmeal at 16, 22, and 28% moisture content were 140.4, 18.9, and 3.3 min at 60 °C, 78.4, 7.1, and 1.6 min at 64 °C, and 37.3, 2.8, and 0.8 min at 68 °C, respectively. The z-values of E. faecium and Salmonella in cornmeal at 16, 22, and 28% moisture content were 13.9, 9.7, and 12.5 °C, and 14.0, 10.4, and 11.7 °C, respectively. These results indicated similar or higher thermal resistance (D-values) and equivalent thermal sensitivity (z-values) of E. faecium compared to Salmonella at different moisture contents and respective temperatures (P ≤ 0.05). Therefore, E. faecium could be used as a surrogate for Salmonella during thermal process validation of cornmeal processing.

Survival and thermal resistance of Salmonella in dry and hydrated nonfat dry milk and whole milk powder during extended storage.

Foodborne pathogens such as Salmonella can endure dry environments of milk powders for extended periods due to the increased adaptability at a low water activity (aw) and proliferate when powders are hydrated. This study compared the survivability and the thermal resistance of a 5-serovar Salmonella cocktail in dry and hydrated nonfat dry milk (NFDM) and whole milk powder (WMP) stored for 180 days at ambient temperature (~20 °C). This study was designed as two factorial (storage days and milk powder type) randomized complete block design with three replications as blocks. The milk powders were spray inoculated with 5-serovar Salmonella cocktail and dried back to the original pre-inoculation aw. The D-values of Salmonella in inoculated NFDM and WMP were determined periodically (every 30 days, starting from day one). The milk powders were also individually hydrated on each analysis day to determine D- and z-values of Salmonella in hydrated powders. The D-values were determined using thermal-death-time disks and hot-water baths at 80, 85 and 90 °C for milk powders, and 59, 62 and 65 °C for hydrated powders. The D- and z-values of Salmonella at specific temperatures within dry or hydrated powders during the storage period were compared at P ≤ 0.05 using two-way ANOVA and Tukey's Test. The D-values of Salmonella in WMP on day 1 were 18.9, 9.9 and 4.4 min at 80, 85 and 90 °C, respectively, which increased to 29.4, 13.6 and 6.5 min at 80, 85 and 90 °C, respectively, on day 180. Whereas, D-values of Salmonella in NFDM on day 1 were 17.9, 9.2 and 4.4 min at 80, 85 and 90 °C, respectively, and stayed similar during the storage. The D-values of Salmonella in milk powder remained similar throughout the storage once hydrated. The overall z-value of Salmonella in NFDM and WMP was 16.3 °C, whereas in hydrated NFDM and WMP, the overall z-value was 6.4 °C.