Adaptive Laboratory Evolution of Probiotics toward Oxidative Stress Using a Microfluidic-Based Platform.

Adaptive laboratory evolution (ALE) can be used to make bacteria less susceptible to oxidative stress. An alternative to large batch scale ALE cultures is to use microfluidic platforms, which are often more economical and more efficient. Microfluidic ALE platforms have shown promise, but many have suffered from subpar cell passaging mechanisms and poor spatial definition. A new approach is presented using a microfluidic Evolution on a Chip (EVoc) design which progressively drives microbial cells from areas of lower H2O2 concentration to areas of higher concentration. Prolonged exposure, up to 72 h, revealed the survival of adaptive strains of Lacticaseibacillus rhamnosus GG, a beneficial probiotic often included in food products. After performing ALE on this microfluidic platform, the bacteria persisted under high H2O2 concentrations in repeated trials. After two progressive exposures, the ability of L. rhamnosus to grow in the presence of H2O2 increased from 1 mm H2O2 after a lag time of 31 h to 1 mm after 21 h, 2 mm after 28 h, and 3 mm after 42 h. The adaptive strains have different morphology, and gene expression compared to wild type, and genome sequencing revealed a potentially meaningful single nucleotide mutation in the protein omega-amidase.

Survival of Salmonella enterica in Military Low-Moisture Food Products during Long-Term Storage at 4, 25, and 40°C.

ABSTRACT: Salmonella enterica has been increasingly implicated in foodborne outbreaks involving low-moisture foods (LMF) during the recent decade. This study aimed to investigate the potential for persistence of S. enterica in a range of LMF during storage at three temperatures. LMF products, boil-in-bag eggs (freeze-dried product), chocolate protein drink, cran-raspberry First Strike bars, mocha dessert bar, and peanut butter, were inoculated with a five-strain cocktail of S. enterica and stored at 4, 25, or 40°C for 36 months. Salmonella populations remained above 7 log CFU/g in all products stored at 4°C and above 6 log CFU/g in products stored at 25°C, excluding the cran-raspberry First Strike bars. Storage at 40°C resulted in Salmonella populations above 5.5 log CFU/g in boil-in-bag eggs after 36 months and demonstrated survivability for 12 months or less in the other five products. Additionally, a mocha bar production temperature profile study identified rapid cooling of bars in which the temperatures reached would have no measurable impact on Salmonella populations. The results indicate the ability of Salmonella to survive in a variety of LMF category foods, even under adverse storage conditions and identifies how the food matrix may affect Salmonella survivability. The data indicate the importance of establishing food processing procedures that adequately mitigate the presence of Salmonella throughout food processing systems, while also increasing comprehensive understanding of Salmonella survivability mechanisms.

Ultrasonic agglomeration of model flour systems: Process parameter-product physico-thermal property relationships.

Ultrasonic compression was applied to wheat flour to create an agglomerated and compacted model food system. This novel process combines physical compression with ultrasonic vibration to permanently weld particles together, thereby producing a robust compact. Fundamental relationships among operating parameters, energy imparted to the specimens, and physical properties of the agglomerated products were developed. Integrated agglomeration energy was determined to be a linear function of percent maximum horn amplitude and ultrasonication time, and integrated energy in turn influenced product physical properties, such as density and fracture strength, in highly significant linear relationships. Microscopic, computer tomographic, and differential scanning calorimetric analyses confirmed progressive compaction of and thermal stability changes in the flour matrix with increased processing energy. Specimens agglomerated at higher energy levels were furthermore demonstrated to have higher fracture strength than conventionally (pressure-only) compressed specimens despite similar densities, due to the robust interparticle bridging produced by agglomeration. PRACTICAL APPLICATION: Ultrasonic agglomeration represents a potential improvement over standard compression in the manufacturing of meal-supplementing energy bars. Ultrasonic agglomeration effectively adheres particles together without incorporation of low-nutrient- density syrups and binders. Agglomerated and compacted products also have superior mechanical stability, which can improve texture and physical stability during product handling.

Efficacy of a Sonicating Swab for Removal and Capture of Microorganisms from Experimental and Natural Contaminated Surfaces.

Enhancements in swabbing technology to increase sample collection efficacy would benefit the food industry. Specifically, these enhancements would assist the food industry in implementing the FDA Food Safety Modernization Act (FSMA) requirements by improving environmental monitoring effectiveness. A sonicating swab device, an example of an enhanced swabbing technology, was demonstrated previously to remove biofilm from stainless steel more efficiently than a standard cotton swab. Within this study, the performance of the sonicating swab was compared to that of the standard cotton swab for the recovery of Listeria monocytogenes from inoculated surfaces (plastic cutting board, wood cutting board, vinyl floor tile, and quarry clay floor tile). Additionally, we demonstrate the sonicating swab performance for collection of a microbiological sample from used commercial plastic cutting boards (noninoculated) in comparison to cotton swabs, foam swabs, and sponges. The sonicating swab captured significantly (P ≤ 0.05) more L. monocytogenes than the cotton swab for both the quarry tile and wood cutting board, while no significant differences were observed for the plastic cutting board or the vinyl floor tile. The sonicating swab consistently recovered significantly (P ≤ 0.05) more bacteria from the used cutting boards than did the standard cotton swab or the 3M Enviro swab, and it recovered significantly (P ≤ 0.05) more bacteria than the sponge swab for a majority of the time (4 of 6 trials). The results of this study indicate that swab technology can still be improved and that the sonicating swab is a viable technological enhancement which aids microbiological sample collection.IMPORTANCE Swabbing of surface areas for microbial contamination has been the standard for the detection and enumeration of microorganisms for many years. Inadequate surface sampling can result in foodborne illness outbreaks due to biotransfer of harmful microorganisms from food contact surfaces to foods. Swab material type, surface characteristics, and swabbing method used are a few of the factors associated with swabbing that can result in the variability of bacterial cell recovery for detection and enumeration. A previous study highlighted a sonicating swab prototype and its ability to recover cells from a stainless steel surface more efficiently and reliably than a standard swab method (T. A. Branck, M. J. Hurley, G. N. Prata, C. A. Crivello, and P. J. Marek, Appl Environ Microbiol 83:e00109-17, 2017, https://doi.org/10.1128/AEM.00109-17). This study expands upon the capabilities of the sonicating swab technology to recover cells from multiple surface types with increased performance over traditional swabbing methods as a tool to further assist in the prevention of foodborne illness outbreaks.