Metagenomics of pasteurized and unpasteurized gouda cheese using targeted 16S rDNA sequencing.

BACKGROUND: The microbiome of cheese is diverse, even within a variety. The metagenomics of cheese is dependent on a vast array of biotic and abiotic factors. Biotic factors include the population of microbiota and their resulting cellular metabolism. Abiotic factors, including the pH, water activity, fat, salt, and moisture content of the cheese matrix, as well as environmental conditions (temperature, humidity, and location of aging), influence the biotic factors. This study assessed the metagenomics of commercial Gouda cheese prepared using pasteurized or unpasteurized cow milk or pasteurized goat milk via 16S rDNA sequencing. RESULTS: Results were analyzed and compared based on milk pasteurization and source, spatial variability (core, outer, and under the rind), and length of aging (2-4 up to 12-18 months). The dominant organisms in the Gouda cheeses, based on percentage of sequence reads identified at the family or genus levels, were Bacillaceae, Lactococcus, Lactobacillus, Streptococcus, and Staphylococcus. More genus- or family-level (e.g. Bacillaceae) identifications were observed in the Gouda cheeses prepared with unpasteurized cow milk (120) compared with those prepared with pasteurized cow milk (92). When assessing influence of spatial variability on the metagenomics of the cheese, more pronounced differences in bacterial genera were observed in the samples taken under the rind; Brachybacterium, Pseudoalteromonas, Yersinia, Klebsiella, and Weissella were only detected in these samples. Lastly, the aging length of the cheese greatly influenced the number of organisms observed. Twenty-seven additional genus-level identifications were observed in Gouda cheese aged for 12-18 months compared with cheese only aged 2-4 months. CONCLUSIONS: Collectively, the results of this study are important in determining the typical microbiota associated with Gouda cheese and how the microbiome plays a role in safety and quality.

Control of Listeria monocytogenes in Caramel Apples by Use of Sticks Pretreated with Potassium Sorbate.

A multistate listeriosis outbreak associated with caramel apples from 2014 to 2015 prompted research on the survival of Listeria monocytogenes in fresh apples and caramel apples. Research indicated that stem end-inoculated caramel apples with stick insertion allowed for the survival and growth of L. monocytogenes at both refrigeration and ambient temperatures. This study aimed to assess the effectiveness of chemical preservatives as pretreatments for the wooden stick component to reduce L. monocytogenes loads in stem end-inoculated caramel apples during storage. Wooden sticks were pretreated with 1, 3, or 5% ascorbic acid (vitamin C), Nisaplin (2.5% nisin), potassium sorbate, and sodium benzoate and then inoculated with L. monocytogenes at 7 log CFU per stick. After storage at 25°C, the pathogen was reduced most effectively by the ascorbic acid pretreatments. At all three ascorbic acid concentrations tested, L. monocytogenes levels were reduced below the level of enumeration (2.5 log CFU per apple) at 24 h and were no longer detectable by enrichment after 72 h. Ascorbic acid (5, 10, and 20%) and potassium sorbate (10, 20, 30, and 40%) were further tested as wooden stick pretreatments for pathogen reduction on stem end-inoculated caramel apples stored at 5 and 25°C. The 40% potassium sorbate solution at 25°C was the most effective pretreatment condition in caramel apples and demonstrated a 3.1-log CFU per apple overall decrease in L. monocytogenes population levels after 216 h. Pretreatment of the wooden stick component of a caramel apple with potassium sorbate may be a viable preventive measure to reduce postprocess L. monocytogenes population levels and hence reduce consumer risk associated with caramel apple consumption.

Listeria monocytogenes Growth Kinetics in Milkshakes Made from Naturally and Artificially Contaminated Ice Cream.

This study assessed the growth of Listeria monocytogenes in milkshakes made using the process-contaminated ice cream associated with a listeriosis outbreak in comparison to milkshakes made with artificially contaminated ice cream. For all temperatures, growth kinetics including growth rates, lag phases, maximum populations, and population increases were determined for the naturally and artificially derived contaminants at 5, 10, 15, and 25°C storage for 144 h. The artificially inoculated L. monocytogenes presented lower growth rates and shorter lag phases than the naturally contaminated populations at all temperatures except for 5°C, where the reverse was observed. At 25°C, lag phases of the naturally and artificially contaminated L. monocytogenes were 11.6 and 7.8 h, respectively. The highest increase in population was observed for the artificially inoculated pathogen at 15°C after 96 h (6.16 log CFU/mL) of storage. Growth models for both contamination states in milkshakes were determined. In addition, this study evaluated the antimicrobial effectiveness of flavoring agents, including strawberry, chocolate and mint, on the growth of the pathogen in milkshakes during 10°C storage. All flavor additions resulted in decreased growth rates of L. monocytogenes for both contamination states. The addition of chocolate and mint flavoring also resulted in significantly longer lag phases for both contamination states. This study provides insight into the differences in growth between naturally and artificially contaminated L. monocytogenes in a food product.

Fate of Listeria monocytogenes in Fresh Apples and Caramel Apples.

An outbreak of listeriosis in late 2014 and early 2015 associated with caramel apples led to questions about how this product became a vector for Listeria monocytogenes. This investigation aimed to determine information about the survival and growth of L. monocytogenes in both fresh apples and caramel apples, specifically examining the effects of site and level of inoculation, inoculum drying conditions, and storage temperature. At a high inoculation level (7 log CFU per apple), L. monocytogenes inoculated at the stem end proliferated on Gala caramel apples at both 5 and 25°C and on Granny Smith caramel apples at 25°C by as much as 3 to 5 log CFU per apple. Fresh apples and caramel apples inoculated at the equatorial surface supported survival but not growth of the pathogen. Growth rates (µmax) for apples inoculated at the stem end, as determined using the Baranyi and Roberts growth model, were 1.64 ± 0.27 and 1.38 ± 0.20 log CFU per apple per day for Gala and Granny Smith caramel apples, respectively, stored at 25°C. At a low inoculation level (3 log CFU per apple), L. monocytogenes inoculated at the stem end and the equatorial surface survived but did not grow on fresh Gala and Granny Smith apples stored at 25°C for 49 days; however, on caramel apples inoculated at the stem end, L. monocytogenes had significant growth under the same conditions. Although certain conditions did not support growth, the pathogen was always detectable by enrichment culture. The inoculation procedure had a significant effect on results; when the inoculum was allowed to dry for 24 h at 5°C, growth was significantly slowed compared with inoculum allowed to dry for 2 h at 25°C. Variation in stick materials did affect L. monocytogenes survival, but these differences were diminished once sticks were placed into caramel apples.