ABSTRACT
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.
ABSTRACT
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.
ABSTRACT
Pathogens, such as Salmonella and Listeria monocytogenes, can survive under the dry environment of flour for extended periods of time and could multiply when flour is hydrated to prepare batter or dough. Therefore, inactivation of these pathogens during the cooking/baking step is vital to ensure the microbiological safety of bakery products such as brownies. The aim of this research was to validate a simulated commercial baking process as a kill-step for controlling Salmonella and L. monocytogenes in brownies and to determine thermal inactivation parameters of these pathogens in brownie batter. Independent studies were conducted in a completely randomized design for each pathogen. All-purpose flour was inoculated with a 5-serovar Salmonella and 3-strain L. monocytogenes cocktails. For baking validation, brownie batters were prepared from inoculated flour, and cooked in the oven set at 350°F (176.7°C) for 40 min followed by 15 min of ambient air cooling. For calculating D-values, brownie batter was transferred into thermal-death-time disks, sealed, and placed in hot-water baths. The samples were held for pre-determined time intervals in hot-water baths and immediately transferred to cold-water baths. Microbial populations were enumerated using injury-recovery media. At the end of baking, Salmonella and L. monocytogenes populations decreased by 6.3 and 5.9 log CFU/g, respectively. D-values of Salmonella and L. monocytogenes cocktails were 53.4 and 37.5 min at 64°C; 27.2 and 16.9 min at 68°C; 10.7 and 9.1 min at 72°C; and 4.6 and 7.3 min at 76°C; respectively. The z-values of Salmonella and L. monocytogenes cocktails were 11.1 and 16.4°C, respectively. This study can be used as a supporting document for the validation of similar brownie baking processes to control Salmonella and L. monocytogenes. The data from this study can also be employed for developing basic prediction models for the survival and thermal resistance of these pathogens during brownie baking step.
ABSTRACT
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.
