Search

Preventing the introduction of Listeria monocytogenes, subsequent biofilm formation, and persistence in food processing environments is important for reducing the risk of cross-contamination of ready-to-eat foods. This study determined the effect of Ɛ-poly-lysine (EPL), hydrogen peroxide (HP), and lauric arginate (LAE) on L. monocytogenes biofilm formation and the inactivation of mature biofilms. For inhibition studies, biofilms of L. monocytogenes Scott A (serotype 4b) and 2014L-6025 (serotype 1/2b) were developed separately at 37 °C for 48 h in the presence of sub-inhibitory concentrations (SIC) of either EPL (10 ppm), HP (2 ppm), or LAE (1.5 ppm) on polystyrene plates and stainless-steel rounds. Inactivation was determined by exposing mature biofilms on each surface to each antimicrobial at their minimum bactericidal concentration (MBC), 10xMBC, or 100xMBC for 24 h at 37 °C. The presence of these antimicrobials at SIC did not inhibit biofilm formation on either surface and their effect on mature biofilms varied by strain and surface. Application of EPL at 1xMBC (100 ppm) for 24 h resulted in greater reductions in counts of both strains on polystyrene than HP (40 ppm) and LAE (5 ppm) under the same conditions at 1xMBC (P ≤ 0.0243). Exposure of mature biofilms to LAE at 10xMBC (50 ppm) for 1 h was more effective in reducing counts on polystyrene than HP at 10xMBC (400 ppm) for the same duration (P ≤ 0.0136), and both HP and LAE applied at 100xMBC (4,000 and 500 ppm, respectively) for 24 h more effectively inactivated mature biofilms of L. monocytogenes Scott A on polystyrene compared to EPL (10,000 ppm) (P ≤ 0.0307). Application of LAE at 10xMBC for 24 h was more effective at inactivating strain Scott A on stainless steel compared to 10xMBC of EPL (1,000 ppm) or HP (P ≤ 0.0430). Future studies are needed to determine the efficacy of these and other antimicrobials on additional strains and serotypes of L. monocytogenes at temperatures relevant to food production and storage.

Recent studies on the use of plant-derived and other bioactive compounds and antimicrobials in food have challenged the idea that exposure to antimicrobials at sublethal or subinhibitory concentrations (SICs) increases the virulence potential of bacterial pathogens including Listeria monocytogenes. The objective of this study was to determine the effect of exposure to SICs of Ɛ-polylysine (EPL), hydrogen peroxide (HP), and lauric arginate (LAE) on L. monocytogenes virulence. For all assays, L. monocytogenes strains Scott A and 2014L-6025 were grown to mid-log phase in the presence of SICs of EPL, HP, or LAE. Motility was determined by spot inoculating cultures on soft brain heart infusion agar (0.3% agar). Cultures grown in SICs of antimicrobials were also inoculated onto Caco-2 cells (10:1 MOI) to determine the effects on subsequent adhesion and invasion. Last, the relative expression of key virulence genes (prfA, plcB, hlyA, actA, inlA, inlB, sigB, and virR) following growth in SICs was determined by RT-qPCR. Results indicate that L. monocytogenes growth in the presence of SICs of EPL, HP, or LAE did not affect the motility, adhesion, or invasion capacity of either strain. Changes in gene expression were observed for both L. monocytogenes strains. More specifically, SICs of EPL and LAE reduced hlyA expression in Scott A, whereas SICs of EPL and HP increased the expression of virR. The upregulation of sigB and actA in the presence of EPL and LAE, respectively, was observed in strain 2014L-6025. These findings indicate that exposure to SICs of these antimicrobials has varying effects on L. monocytogenes that differ by strain. Although no phenotypic effects were observed in terms of motility, adhesion, and invasion, the observed changes in virulence gene expression warrant further investigation.

Microalgae, known for their land-sparing cultivation and remarkable photosynthetic efficiency, play a crucial role in advancing sustainable development goals. This study explored the use of cost-effective inorganic carbon (the ionic form of CO2) and organic carbon sources (lactose, a primary sugar in whey wastewater) to support the growth of Chlorella sorokiniana UTEX 1230, and improved protein content through nitrogen supply optimization. NaHCO3 proved to be more effective than Na2CO3, resulting in a 3.4-fold increase in biomass concentration, with an initial 5 mM HCO3‾ concentration compared to the basal medium. Further addition of 0.75 % lactose led to a 4.05-fold increase in biomass concentration, with the maximum specific growth rate at 1.28/d, maximum biomass concentration at 695.88 mg/L, and maximum biomass productivity at 250.91 mg/L/d. Nitrogen supplementation greatly increased protein concentration from 16.41 % (250 mg/L NaNO3) to 52.95 % (1000 mg/L NaNO3). These results support the potential for utilizing whey wastewater bioremediation to produce high-value protein via microalgal cultivation.

ABSTRACT Prior to the advent of milk pasteurization and the use of defined-strain starter cultures, the production and ripening of cheese relied on the introduction and growth of adventitious microbes from the environment. This study characterized microbial community structures throughout a traditional farmstead cheese production continuum and evaluated the role of the environment in microbial transfer. In total, 118 samples (e.g., raw milk, cheese, and environmental surfaces) were collected from milk harvesting through cheese ripening. Microbial communities were characterized based on amplicon sequencing of bacterial 16S rRNA and fungal internal transcribed spacer genes using the Illumina MiSeq platform. Results indicated that the environment in each processing room harbored unique microbial ecosystems and consistently contributed microbes to milk, curd, and cheese. The diverse microbial composition of milk was initially attributed to milker hands and cow teats and then changed substantially following overnight ripening in a wooden vat to one dominated by lactic acid bacteria, including Lactococcus lactis, Lactobacillus, and Leuconostoc, as well as fungi such as Exophiala, Kluyveromyces, and Candida. Additional microbial contributions were attributed to processing tools, but the composition of the cheese paste remained relatively stable over 60 days of ripening. In contrast, rind communities that were largely influenced by direct contact with bamboo aging mats showed a distinct succession pattern compared to the interior sections. Overall, these findings highlight the critical role of traditional tools and practices in shaping the microbial composition of cheese and broaden our understanding of processing environments as important sources of microbes in food. IMPORTANCE Throughout the 20th century, especially in the United States, sanitation practices, pasteurization of milk, and the use of commercial defined-strain starter cultures have enhanced the safety and consistency of cheese. However, these practices can reduce cheese microbial diversity. The rapid growth of the artisanal cheese industry in the United States has renewed interest in recapturing the diversity of dairy products and the microbes involved in their production. Here, we demonstrate the essential role of the environment, including the use of wooden tools and cheesemaking equipment, as sources of dominant microbes that shape the fermentation and ripening processes of a traditional farmstead cheese produced without the addition of starter cultures or direct inoculation of any other bacteria or fungi. These data enrich our understanding of the microbial interactions between products and the environment and identify taxa that contribute to the microbial diversity of cheese and cheese production.

Throughout the 20th century, especially in the United States, sanitation practices, pasteurization of milk, and the use of commercial defined-strain starter cultures have enhanced the safety and consistency of cheese. However, these practices can reduce cheese microbial diversity.