Environmental pH and compound structure affect the activity of short-chain carboxylic acids against planktonic growth, biofilm formation, and eradication of the food pathogen Salmonella enterica.

Short-chain carboxylic acids (SCCAs) that are naturally produced by microbial fermentation play an essential role in delaying microbial spoilage. SCCAs are structurally diverse, but only a few of them are routinely used in food biopreservation. This study investigated the effects of environmental pH and intrinsic properties of 21 structurally different SCCAs on the antimicrobial and antibiofilm activity against Salmonella enterica. Inhibition of SCCA toward planktonic and biofilm growth of S. enterica was higher in an acidic environment (pH 4.5) that is common in fermented products, and for SCCA that possessed both a high acid dissociation strength (pKa) (>4.0) and a positive hydrophobicity [octanol/water partition coefficient (log Kow)]. Crotonic and caproic acids were identified as SCCAs with potential as biopreservatives even at near-neutral pH. SCCA with hydrophilic groups such as lactic acid did not inhibit S. enterica at concentrations up to 50 mM, while SCCA with benzene or methyl groups or a double bond prevented S. enterica growth and biofilm formation. Stimulation of biofilm formation was observed for formic, acetic, and propionic acid close to the minimum inhibitory concentration to reduce 50% of cell density (MIC50) of planktonic cells, and for citric and isocitric acid with an MIC50 of ≥50 mM. The presence of low concentrations of formic and propionic acids during biofilm formation conferred protection during eradication possibly due to a pre-adaptation effect, yet two consecutive acid treatments were successful in eradicating biofilms if the first acid treatment was two- to threefold of the MIC50.IMPORTANCEThis study provides a systematic comparison on the antimicrobial and antibiofilm activity of more than 20 structurally different SCCAs against a common food pathogen. We tested the antimicrobial activity at controlled pH and identified the structure-dependent antimicrobial effects of SCCA without the confounding influence of acidification. The combined effect of pKa and log Kow was identified as an important feature that should be considered when deciding for a specific SCCA in the application as antimicrobial. Our results imply that additional phenomena such as the use of SCCA as substrate and cellular pre-adaption effects have to be taken into consideration. We finally present a two-step treatment as an efficient approach to eradicate biofilms, which can be applied for the disinfection of contact surfaces and manufacturing equipment. Results obtained here can serve as guidelines for application of SCCA to avoid the growth of food pathogens and/or to develop biopreserved food systems.

Alginate coatings applied on apple cubes as a vehicle for Lacticaseibacillus casei: probiotic viability and overall quality of a new functional product.

BACKGROUND: Worldwide, vegan and vegetarian lifestyles, as well as food allergies and intolerance (e.g. lactose intolerance and milk protein allergy) demand the development of alternatives to dairy-based probiotic foods. In the present study, probiotic Lacticaseibacillus casei CECT 9104 was added to alginate-based edible coatings enriched with inulin and oligofructose and applied to fresh-cut apple. Microbiological, physicochemical and sensory quality parameters of the apple cubes were monitored during 8 days of refrigerated storage. Lacticaseibacillus casei was tested for its antagonistic effect against inoculated Listeria innocua and Escherichia coli O157:H7. The viability of the probiotic strain during refrigerated storage and after simulated gastrointestinal digestion (GID) was evaluated. RESULTS: After 8 days of storage, 9.52-9.64 log colony-forming units (CFU) g-1 of L. casei were detected in apple samples. The functional apple cubes retained 8.31-8.43 log CFU g-1 of the probiotic after GID, without a significant effect of prebiotic addition. The microbiological quality and nutritional properties were maintained by the use of active coatings, whereas the sensory quality decreased after 8 days of storage. A bactericidal effect was exerted by the probiotic strain loaded in the coating against L. innocua artificially inoculated on apple cubes. Escherichia coli O157:H7 counts were reduced by 2.5 log after 8 days. CONCLUSION: This study has demonstrated the suitability of apple cubes as an alternative matrix to milk for carrying probiotic L. casei CECT 9104 and prebiotics, offering a promising alternative for the development of plant-based functional foods. © 2024 Society of Chemical Industry.

Novel Lactobacillaceae strains and consortia to produce propionate-containing fermentates as biopreservatives.

Biopreservation refers to the use of natural or controlled microbial single strains or consortia, and/or their metabolites such as short-chain carboxylic acids (SCCA), to improve the shelf-life of foods. This study aimed at establishing a novel Lactobacillaceae-driven bioprocess that led to the production of the SCCA propionate through the cross-feeding on 1,2-propanediol (1,2-PD) derived from the deoxyhexoses rhamnose or fucose. When grown as single cultures in Hungate tubes, strains of Lacticaseibacillus rhamnosus preferred fucose over rhamnose and produced 1,2-PD in addition to lactate, acetate, and formate, while Limosilactobacillus reuteri metabolized 1,2-PD into propionate, propanol and propanal. Loigolactobacillus coryniformis used fucose to produce 1,2-PD and only formed propionate when supplied with 1,2-PD. Fermentates collected from batch fermentations in bioreactor using two-strain consortia (L. rhamnosus and L. reuteri) or fed-batch fermentations of single strain cultures of L. coryniformis with rhamnose contained mixtures of SCCA consisting of mainly lactate and acetate and also propionate. Synthetic mixtures that contained SCCA at concentrations present in the fermentates were more antimicrobial against Salmonella enterica if propionate was present. Together, this study (i) demonstrates the potential of single strains and two-strain consortia to produce propionate in the presence of deoxyhexoses extending the fermentation metabolite profile of Lactobacillaceae, and (ii) emphasizes the potential of applying propionate-containing fermentates as biopreservatives.

Effects of combined application of high-pressure processing and active coatings on phenolic compounds and microbiological and physicochemical quality of apple cubes.

BACKGROUND: In recent years the use of high-pressure processing (HPP) of fruit products has steadily increased due to its antimicrobial effectiveness and the retention of nutritional and quality attributes compared to conventional thermal technologies. Edible coatings are already being used to enhance the quality of minimally processed fruits. Thus, apple cubes (AC) and alginate-vanillin-coated apple cubes (AVAC) were subjected to HPP (400 MPa/5 min/35 °C). The microbiological and physicochemical parameters were evaluated and the bioactive compounds were monitored before and after HPP of apple cubes. Also, an in vitro gastrointestinal digestion (GID) was conducted. RESULTS: HPP left L. monocytogenes counts below the detection limit (2 log UFC g-1 ), regardless of the presence of coating. For E. coli, HPP + active coating showed a synergism affording the greatest reduction (>5 log) for AVAC-HPP. Firmness was maintained in AVAC-HPP samples, while AC-HPP samples suffered reductions of 35%. Colour attributes were also better retained in AVAC-HPP samples. In general, HPP led to a decrease in phenolic compounds. Regarding the effects of GID, vanillin-based active coating exerted a protective effect on some phenolics. Thus, p-coumaroylquinic acid concentration was maintained for AVAC and AVAC-HPP during GID. Epigallocatechin, the compound with the highest concentration in apple cubes, increased for AVAC (106%) and AVAC-HPP (57%). Also, phloridzin concentration increased for AVAC-HPP (17%). At the end of GID, procyanidin B1 and epigallocatechin were the main phenolic compounds for all samples, AVAC showing the highest concentration. CONCLUSIONS: This work demonstrates that the combined application of HPP and active coatings on apple cubes could be used to obtain a safe and good-quality product. © 2021 Society of Chemical Industry.

Novel functional blueberries: Fructo-oligosaccharides and probiotic lactobacilli incorporated into alginate edible coatings.

Minimally processed fruits are an alternative to dairy products to deliver probiotics. Bio-protection against several factors that affect their viability has been proposed in the food industry. In this study, probiotic Lactobacillus rhamnosus CECT 8361 was added to alginate-based coatings enriched with inulin and oligofructose and applied on fresh-blueberries. Probiotic viability, microbiological, physicochemical and sensory quality parameters of blueberries were monitored during 21 days of refrigerated storage. Also, L. rhamnosus CECT 8361 was tested for its antagonistic effect against inoculated Listeria innocua and E. coli O157:H7. Advantageously, prebiotic compounds allowed improving probiotic viability with counts above 6.2 log CFU/g for the entire period. Native microbiota counts remained under safe levels. Overall visual quality, odor and flavor were acceptable up to day 14 of storage. Regarding antimicrobial activity, L. rhamnosus CECT 8361 was able to reduce L. innocua counts by 1.7 log in inoculated blueberries. These results encourage further implementation of new fruit-based foods with multifunctional properties.