Multi-functional polyvinyl alcohol/tannin acid composite films incorporated with lignin nanoparticles loaded by potassium sorbate.

The biocompatible, biodegradable and strong polyvinyl alcohol-based films have been widely investigated and used in the field of active packaging. To endow with diverse function, this paper firstly prepared lignin nanoparticles loaded with potassium sorbate (LNP@PS) as additives to exploit additional antibacterial, UV blocking, oxygen barrier, and water barrier properties. Besides, tannin acid (TA) was incorporated for compensating and further enhancing mechanical properties. Results showed that the PVA-based composite films containing 3 % LNP@PS and 5 % TA could achieve the optimal tensile strength at 74.51 MPa, water vapor permeability at 7.015·10-13·g·cm/cm2·s·Pa and oxygen permeability at 1.93 cm3/m2·24 h MPa, which was an 165 % of increase, 47 % and 112 % of reduction respectively compared to pure PVA films. Additionally, the composite films exhibited apparently superior bacteria and oxygen resistance properties evidenced by microbial infection and free radical scavenging performance. In addition, the slow-release effect of PS assisted the strawberry preservation with an extension of 3 days, which provided a promising novel route to prepare active food packaging material.

Utilizing functional genomics in Saccharomyces cerevisiae to characterize food preservative compounds: A pilot study.

Chemical preservatives are ubiquitously used to suppress the growth of or kill microorganisms across numerous industries, including the food industry. Utilizing yeast functional genomic techniques, genes and their functions can be observed at a genomic scale to elucidate how environmental stressors (e.g., chemical preservatives) impact microbial survival. These types of chemical genomics approaches can reveal genetic mutations that result in preservative resistance or sensitivity, assist in identification of preservative mechanism of action, and can be used to compare different preservatives for rational design of preservative mixtures. In this proof-of-concept study, we performed deletion and high-copy genetic expression screens to identify mutants that confer drug resistance to sodium benzoate, potassium sorbate, rosemary extract, and Natamax. By observing overlapping mutant genes between genetic screens, we were able to identify functional overlap between chemical preservatives and begin to explain mechanisms of action for these compounds.

Effects of potassium sorbate on systemic inflammation and gut microbiota in normal mice: A comparison of continuous intake and washout period.

Potassium sorbate (PS) is a widely used food preservative in the field of food industry. However, the effects of continuous intake and washout period of PS on host health are still unclear. In this study, to investigate long-term effect and after-effect of different concentrations and time points of PS, healthy mice were orally exposed to 150 mg/kg, 500 mg/kg and 1000 mg/kg of PS for 10 weeks, and washout treatment for another 5 weeks, respectively. The results indicated that PS intake for 10 weeks had no obvious effects on organs and adipose tissue, nor did it noteworthily interfere with glucolipid metabolism in the serum. However, it caused inflammatory cell infiltration in the liver, increased serum interleukin (IL)-1ß level, changed abundances of gut microbiota but failed to promote the production of short chain fatty acids in the gut. After washout period for 5 weeks, liver inflammation and IL-1ß level were decreased, and gut environment developed towards a healthier condition. Specifically, PS washout significantly increased abundance of Lachnospiraceae_NK4A136_group and the production of isobutyric acid. This study confirmed washout period eliminated negative effects from continuous intake of PS, which provided positive evidence for its safety.

Inactivation of Bacillus subtilis spores by a combination of high-pressure thermal treatment and potassium sorbate.

Combining High-pressure Thermal Treatment (HPTT) and Potassium Sorbate (PS) may have a stronger spore inactivation effect. Spores of Bacillus subtilis were subjected to HPTT at 600 MPa-65 °C/75 °C and a combination of HPTT and PS of 0.1% and 0.2% concentrations. After these treatments, different procedures and techniques were employed to investigate the spore's inactivation. The results revealed that 4.92 ± 0.05 log spores were inactivated after treatment at 600 MPa-75 °C, while 5.97 ± 0.09 log spores were inactivated when the HPTT treatment was combined with 0.2% PS. Changes in permeability of the spore's inner membrane were characterized by OD600 value and release rates of nucleic acids, protein, and dipicolinic acid (DPA). Compared with HPTT treatment at 600 MPa-75 °C, the OD600 value of spores decreased further by about 50% after treatment with a combination of HPTT and 0.2% PS. Additionally, the combined treatments resulted in a significant increase in the OD260 and OD280 values, as well as the DPA release. The spore size analysis indicated a significant decrease in the size of spores treated with a combination of HPTT at 600 MPa-75 °C and PS of 0.2% concentration. Furthermore, the flow cytometry analysis and confocal laser scanning microscopy (CLSM) analysis indicated that the inner membrane damage of spores was higher after combined treatments than that after HPTT treatment alone. A significant reduction was also found in the Na+/K+-ATPase activity after the combined treatments. Also, the FTIR analysis revealed that the combined treatments resulted in significant adverse changes in the spores' inner membrane, cell wall, cortex, and nucleic acid. Therefore, the combination of HPTT and PS has a stronger inactivation effect and can be suggested as a promising strategy for the inactivation of Bacillus subtilis spores.

Evaluating the potential of natural product combinations with sorbic acid for improving preservative action against food-spoilage yeasts.

Fungal control methods commonly involve the use of antifungals or preservatives, which can raise concerns about broader effects of these stressors on non-target organisms, spread of resistance and regulatory hurdles. Consequently, control methods enabling lower usage of such stressors are highly sought, for example chemical combinations that synergistically inhibit target-organisms. Here, we investigated how well such a principle extends to improving efficacy of an existing but tightly controlled food preservative, sorbic acid. A screen of ∼200 natural products for synergistic fungal inhibition in combinations with sorbic acid, in either 2% or 0.1% (w/v) glucose to simulate high or reduced-sugar foods, did not reveal reproducible synergies in either of the spoilage yeast species Saccharomyces cerevisiae or Zygosaccharomyces bailii. Potentially promising screen candidates (e.g. lactone parthenolide, ethyl maltol) or a small additional panel of rationally-selected compounds (e.g. benzoic acid) all gave Fractional Inhibitory Concentration Indices (FICI) ≥ 0.5 in combinations with sorbic acid, corroborating absence of synergy in either glucose condition (although FICI values did differ between the glucose conditions). Synergies were not achieved either in a tripartite combination with screen candidates or in a soft-drink formulation as matrix. In previous work with other stressors synergy 'hits' have been comparatively frequent, suggesting that sorbic acid could be unusually resistant to forming synergies with other potential inhibitors and this may relate to the weak acid's known multifactorial inhibitory-actions on cells. The study highlights a challenge in developing appropriate natural product or other chemical combinations applicable to food and beverage preservation.

Evaluation of Sodium Benzoate and Potassium Sorbate Preservative Concentrations in Different Sauce Samples in Urmia, Iran.

Sodium benzoate and potassium sorbate are relatively common preservatives used in a wide range of foods including flavoring products like sauces. The potential health risks arising from these preservatives along with the high-consumption rate of these flavoring products worldwide highlight the importance of the quality and safety assurance of these products. So, this study aimed to evaluate the concentrations of these two common preservatives (i.e., sodium benzoate and potassium sorbate) in different sauce samples, including mayonnaise, salad dressings, Caesar sauce, Italian dressing, Ranch dressing, French dressing, using high-performance liquid chromatography (HPLC) and to compare them with the acceptable level of Codex standard. For this purpose, 49 samples, including three to five samples of each type of different brands of sauce samples, were randomly collected from supermarkets in Urmia, Iran. Based on the results, the mean concentrations ± standard deviation of sodium benzoate and potassium sorbate in the collected samples were found to be 249.9 ± 157 and 158.0 ± 131 ppm, respectively, which were lower than the general standard of the Codex Alimentarius and the European legislation. Due to the importance of hazardous side effects of these preservatives for consumers, regular and accurate evaluation of these preservatives in sauces as highly consumed food products is still recommended for consumer safety.

Overcoming lemon postharvest molds caused by Penicillium spp. multiresistant isolates by the application of potassium sorbate in aqueous and wax treatments.

Penicillium digitatum and Penicillium italicum are the main causal agents of postharvest diseases in lemon. Over the last decades, the appearance of isolates resistant to the main commercial fungicides has been considered one of the most serious problems for the citrus industry. In this work, potassium sorbate (KS) was evaluated as an alternative to chemical fungicides to control postharvest diseases caused by Penicillium isolates resistant to imazalil, thiabendazol, and pyrimethanil. In vitro assays showed that 1% KS inhibited conidia germination and mycelial growth of sensitive and resistant P. digitatum and P. italicum isolates, being this effect stronger at pH 5 than at pH 9. In curative treatments, the immersion of inoculated lemons in 1% KS aqueous solution for 30 s reduced green and blue molds incidences by around 80%. No wound protection effect was observed when wounded lemons were immersed in 3% salt solution before inoculation. Noteworthy, the inclusion of KS in a commercial wax coating effectively controlled green and blue molds, even in decays caused by fungicide resistance isolates. Together, results encourage the use of KS in lemon postharvest treatments to contribute to the management of resistant strains, which represent a major challenge in packinghouses worldwide. PRACTICAL APPLICATION: The use of KS in citrus postharvest treatments would help producers to reduce spoilage caused by Penicillium fungicide-resistant strains. The inclusion of this generally recognized as safe compound in wax coatings improves its persistence on the fruit surface, keeping product quality during long-term overseas transport. In sum, KS constitutes an affordable and eco-friendly option for controlling postharvest molds in lemon fruit.

Antimicrobial effect of sorbic acid-loaded chitosan/tripolyphosphate nanoparticles on Pseudomonas aeruginosa.

Sorbic acid-loaded chitosan/tripolyphosphate nanoparticles (SANs) have previously been shown to exert both antibacterial and antioxidant effects on Chinese sausage. In this study, the minimum inhibitory concentrations (MICs) of SANs against two Pseudomonas aeruginosa strains were determined. The blank control group (BC) served as the negative control, while the chitosan/tripolyphosphate nanoparticles (CTNs) group and free sorbic acid (SA) group served as the positive controls. Tests conducted under five different pH conditions (5/6/7/8/9) revealed that the SANs exhibited a good bacteriostatic effect against P. aeruginosa. Variations in the metabolism, cell membrane or cell wall integrity, and morphology of P. aeruginosa were measured to evaluate the effects of SANs on their intracellular and extracellular components. The MIC of SANs for the two P. aeruginosa strains was determined to be 150 µg/mL. SANs delayed the growth of P. aeruginosa and severely damaged both its inner and outer cell membranes. The heteromorphism of the bacteria as observed by field emission scanning electron microscopy (FESEM), verified the aforementioned results. The results showed SANs could effectively inhibit the growth of P. aeruginosa and exert antibacterial ability in a wider range of acid-base environments. This study broadens the application of SANs in food processing and provides experimental basis.

Prevention of microbes-induced spoilage in sodium chloride-free cucumber fermentations employing preservatives.

This study evaluated preservatives to stabilize sodium chloride (NaCl)-free-cucumber fermentations. The brining of air-purged laboratory cucumber fermentations with 100.0 mM calcium chloride (CaCl2 ) and 25.0 mM acetic acid resulted in immediate rises in pH, the chemical reduction of the medium, and malodors. Supplementation with 3.0 mM sodium benzoate or 3.0 mM potassium sorbate enabled a decline in pH, a continuous oxidative state of the medium, and delayed rising pH spoilage. However, lactic and acetic acids eventually disappeared in fermentations supplemented with preservatives. The amount of preservatives needed to suppress growth of brined-cucumber-spoilage microbes was determined in Fermented Cucumber Juice Medium (FCJM). Supplementation of FCJM with 10.0 mM sodium benzoate was inhibitory for the spoilage yeasts, Issatchenkia occidentalis and Pichia manshurica, and the lactobacilli, Lentilactobacillus buchneri and Lentilactobacillus parafarraginis, but not of Zygosaccharomyces globiformis. Potassium sorbate inhibited the spoilage yeasts at 15.0 mM in FCJM but not the lactobacilli. Supplementation of FCJM with 20.0 mM fumaric acid had a bactericidal effect on the spoilage-associated lactobacilli. As expected, NaCl-free-commercial cucumber fermentations brined with 100 mM CaCl2 , no acetic acid, and 6 mM potassium sorbate resulted in complete fermentations, but supported rising pH, microbially induced spoilage during long-term storage. Post-fermentation supplementation with 12 mM sodium benzoate, 10 mM fumaric acid, a combination of the two, or 10 mM fumaric acid and 2 mM AITC prevented microbial activity during long-term bulk storage. PRACTICAL APPLICATION: Several preservative-based strategies for stabilizing NaCl-free cucumber fermentation in a commercial production setting were developed, enabling the implementation of a processing technology that reduces wastewater volumes and environmental impact.

Efficacy of weak acid preservatives on spoilage fungi of bakery products.

Organic acids and their salts are usually the first choice in the bread industry to restrict fungal spoilage, but their efficacy is pH-dependent and spoilage by fungi remains as a common threat. Therefore, this study aimed to evaluate the susceptibility of spoilage fungi of bakery products to acetic, sorbic, and propionic acids at different pH. Penicillium roqueforti, Penicilium paneum, Aspergillus pseudoglaucus, Aspergillus montevidensis and Hyphopichia burtonii strains isolated from spoiled products had their minimum inhibitory concentration (MIC) defined by macrodilution. The concentrations tested were: (i) sorbic acid up to 32 mM; (ii) propionic acid up to 1024 mM and (iii) acetic acid up to 800 mM with pH adjusted in 4.5, 5.0, 5.0 and 6.0 after setting the agent concentration. The lowest MICs for all agents were obtained at pH 4.5, usually doubling with every 0.5 pH increase. P. roqueforti strains isolated from spoiled products were the most resistant to all tested preservatives; while strains of the related species P. paneum, showed similar tolerance to acetic and propionic acids but was double more susceptible to sorbic acid. Strains of A. pseudoglaucus and A. montevidensis were indistinctly susceptible to the preservatives and were the most susceptible species to propionic and acetic acids. H. burtonii strains demonstrated the most variable behaviour in comparison to the other strains being the most susceptible to sorbic acid, were like Aspergillus strains regarding propionic acid, but tolerate well acetic acid. Propionic acid concentrations usually allowed in baked goods are lower than the concentrations required to inhibit the most tolerant isolates tested in this study. The same is true for sorbic acid at higher pH levels. Spoilage species of bakery ware presents a distinct susceptibility profile to the preservatives commonly used in this sector, but the high tolerance observed is a cause of concern.

High sorbic acid resistance of Penicillium roqueforti is mediated by the SORBUS gene cluster.

Penicillium roqueforti is a major food-spoilage fungus known for its high resistance to the food preservative sorbic acid. Here, we demonstrate that the minimum inhibitory concentration of undissociated sorbic acid (MICu) ranges between 4.2 and 21.2 mM when 34 P. roqueforti strains were grown on malt extract broth. A genome-wide association study revealed that the six most resistant strains contained the 180 kbp gene cluster SORBUS, which was absent in the other 28 strains. In addition, a SNP analysis revealed five genes outside the SORBUS cluster that may be linked to sorbic acid resistance. A partial SORBUS knock-out (>100 of 180 kbp) in a resistant strain reduced sorbic acid resistance to similar levels as observed in the sensitive strains. Whole genome transcriptome analysis revealed a small set of genes present in both resistant and sensitive P. roqueforti strains that were differentially expressed in the presence of the weak acid. These genes could explain why P. roqueforti is more resistant to sorbic acid when compared to other fungi, even in the absence of the SORBUS cluster. Together, the MICu of 21.2 mM makes P. roqueforti among the most sorbic acid-resistant fungi, if not the most resistant fungus, which is mediated by the SORBUS gene cluster.

Synergistic activity of Stryphnodendron adstringens and potassium sorbate against foodborne bacteria.

Stryphnodendron adstringens is a medicinal plant that has a broad spectrum of action, including antibacterial activity. The aim of the present study was to evaluate the effect of S. adstringens alone and in combination with potassium sorbate (PS) against foodborne bacteria. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) were determined and, for most of the bacteria tested, the crude extract (CE), aqueous fraction (AQF), and ethyl-acetate fraction (EAF) of S. adstringens had a MIC and MBC ranging from 500 to ≥ 1000 µg/mL. The AQF and EAF showed greater activity against S. aureus strains (MIC = 125 to 250 µg/mL; MBC = 500 to 1000 µg/m). Quantitative cell viability was determined and was observed reductions ranging from 3.0 to 5.8 log10 CFU/ml.The combination of S. adstringens and PS against seven S. aureus isolates was determined by the checkerboard method at neutral and acid pH. In a neutral medium, the AQF + PS combination presented synergistic or additive interactions against six S. aureus strains. The combination of EAF + PS resulted in additive interactions against four bacterial isolates. In an acidic medium, the AQF + PS combination was synergistic or additive against all S. aureus, while EAF + PS presented the same effect against six S. aureus strains S. adstringens showed important antibacterial effects against foodborne S. aureus strains. Moreover, the combination of S. adstringens fractions and PS improved the antibacterial activity compared to the compounds utilized individually. The combined use of these compounds may be an alternative to reduce bacterial food contamination and improve food safety.

Adaptation of Campylobacter field isolates to propionic acid and sorbic acid is associated with fitness costs.

AIMS: To reduce the burden of Campylobacter at different stages of the food chain, recent studies have shown the effectiveness of organic acids as a risk mitigation strategy. However, very little is known about possible adaptation responses of Campylobacter that lead to reduced susceptibility to organic acids. Here we investigated the adaptive responses of Campylobacter field isolates to organic acids and estimated the fitness costs. METHODS AND RESULTS: Exposure of two Campylobacter jejuni and one Campylobacter coli isolate to subinhibitory concentrations of propionic acid or sorbic acid resulted in twofold to fourfold increased minimal inhibitory concentration values for the adapted variants. With one exception, the decreased susceptibility was stable in at least 10 successive subcultures without selection pressure. Growth competition experiments revealed a reduced fitness of adapted variants compared to the wild-type isolates. A linear regression model allowed an estimation of the fitness cost. Growth kinetics experiments showed significantly prolonged lag phases in five of six adapted isolates while there was not a direct correlation in the maximum growth rates compared to the wild-type isolates. CONCLUSIONS: The results of the study showed that a stepwise adaptation of Campylobacter to organic acids is possible, but at the detriment of changes in growth behaviour and reduced fitness. SIGNIFICANCE AND IMPACT OF THE STUDY: The study contributes to the understanding of adaptive responses of Campylobacter to organic acids treatments, for example, as part of risk mitigation strategies.

Efficacy of combinations of lactic acid and potassium sorbate against Listeria monocytogenes in chicken stored under modified atmospheres.

The combined effect of lactic acid and potassium sorbate on the growth of L. monocytogenes on chicken legs packaged under modified atmospheres (MAP) and stored at 4 °C was evaluated. An extended lag phase and a lower maximum growth rate for psychrotrophs and mesophiles was found in those samples packaged in 20%CO2/80%N2 and washed with different combinations of lactic acid and potassium sorbate compared to those non-treated with organic acids. Legs packaged in 20%CO2/80%N2 and washed with 3.75% lactic acid- 3.75% potassium sorbate showed a significant (p < 0.05) reduction in L. monocytogenes compared to untreated chicken legs packaged in MAP, which were approximately 2.63 log units lower in the first ones after 8 days of storage. Moreover, this treatment was the most effective in decreasing the maximum growth rate of L. monocytogenes. The chicken legs packaged in atmospheres containing 20%CO2/80%N2, had an extended shelf life, but these atmospheres were not able to reduce L. monocytogenes, thus underlining the need for preventive measures so as to control this pathogen. The immersion of chicken legs in a solution containing 3.75% lactic acid- 3.75% potassium sorbate can reduce L monocytogenes populations on fresh chicken packaged in a modified atmosphere.

Effect of sorbic acid and dual-purpose inoculants on the fermentation quality and aerobic stability of high dry matter rice straw silage.

AIMS: This study aimed to evaluate the feasibility of sorbic acid (SA) as a silage additive and its effects on fermentation quality and aerobic stability of high dry matter (DM) silage. METHODS AND RESULTS: High DM rice straw was ensiled with distilled water (C), 1 × 106  CFU per gram fresh weight (FW) Lactobacillus plantarum and 1 × 106  CFU per gram FW Lactobacillus buchneri (LP+LB) or SA for 45 days with a subsequent aerobic stability test. After ensiling, LP+LB silage had the highest lactic acid (LA) content and the lowest pH value, whereas SA silage had the highest DM and water-soluble carbohydrate (WSC) contents, and the lowest ethanol and ammonia nitrogen (NH3 -N) contents among all silages (P < 0·001). Compared to C silage, SA significantly (P < 0·01) reduced the counts of yeasts but not lactic acid bacteria (LAB). During 6-day aerobic exposure, the continuous pH increase and LA decrease were observed in C and LP+LB silages, and there was no significant change in pH, DM, NH3 -N and WSC contents of SA silage over the whole aerobic exposure. The SA addition slowed the decline of LA and acetic acid (AA) contents as well as the growth of yeasts and aerobic bacteria under aerobic exposure. CONCLUSION: In this study, L. buchneri could not function in high DM rice straw silage while SA effectively improved both the fermentation quality and aerobic stability. SIGNIFICANCE AND IMPACT OF THE STUDY: The SA was more effective than dual-purpose inoculants to improve the aerobic stability of high DM rice straw silage. Thus, SA can be served as a potential antifungal additive for silage with high DM.

Inhibition of the lemon brown rot causal agent Phytophthora citrophthora by low-toxicity compounds.

BACKGROUND: Phytophthora spp., soil-borne oomycetes, cause brown rot (BR) on postharvest lemons. The management of this disease is based on cultural practices and chemical control using inorganic salts of limited efficacy. In the search for new alternatives, the aim of this work was to evaluate the effect of low-toxicity compounds to inhibit the growth of P. citrophthora and to control BR disease on lemons. Sodium bicarbonate, potassium sorbate, polyhexamethylene guanidine, Ascophyllum nodosum extract and a formulation containing phosphite salts plus A. nodosum (P+An) were evaluated. RESULTS: All tested products inhibited mycelial growth, sporangia formation and zoospore germination of P. citrophthora in vitro. In postharvest applications on artificially inoculated lemons, only P+An exhibited a BR curative effect, with incidence reduction of around 60%. When this formulation was applied in field treatments, BR incidence was reduced by 40% on lemons harvested and inoculated up to 30 days post application. CONCLUSION: Our results demonstrate the in vitro direct anti-oomycete effect of low-toxicity compounds and the in vivo efficacy of P+An formulation to control BR, encouraging the incorporation of the latter in the management of citrus BR. © 2020 Society of Chemical Industry.

Nature-Identical Compounds and Organic Acids Ameliorate and Prevent the Damages Induced by an Inflammatory Challenge in Caco-2 Cell Culture.

Bioactive compounds, such as organic acids (OA) and nature-identical compounds (NIC), can exert a role in the protection of intestinal mucosa functionality due to their biological properties. The aim of this study was to understand the role of 2 OA (citric and sorbic acid) and 2 NIC (thymol and vanillin), alone or combined in a blend (OA + NIC), on intestinal barrier functionality, either during homeostatic condition or during an inflammatory challenge performed with pro-inflammatory cytokines and lipopolysaccharides (LPS). The study was performed on the human epithelial cell line Caco-2, a well-known model of the intestinal epithelial barrier. The results showed how OA and NIC alone can improve transepithelial electrical resistance (TEER) and mRNA levels of tight junction (TJ) components, but OA + NIC showed stronger efficacy compared to the single molecules. When an inflammatory challenge occurred, OA + NIC blend was able both to ameliorate, and prevent, damage caused by the pro-inflammatory stimulus, reducing or preventing the drop in TEER and improving the TJ mRNA expression. The data support the role of OA + NIC in modulating gut barrier functionality and reducing the negative effects of inflammation in intestinal epithelial cells, thereby supporting the gut barrier functionality.

Antimicrobial activity of organic acids against Campylobacter spp. and development of combinations-A synergistic effect?

Contaminated poultry meat is considered to be the main source of human infection with Campylobacter spp., a pathogen that asymptomatically colonizes broiler chickens during fattening and contaminates carcasses during slaughter. To prevent or reduce the colonization of broiler flocks with Campylobacter spp., applying different organic acids, especially in combinations, via feed or drinking water seems to be a promising approach. However, only very few combinations of organic acids have been tested for their antibacterial efficacy against Campylobacter spp. Therefore, the in vitro susceptibility of 30 Campylobacter spp. isolates (20 C. jejuni and ten C. coli) to ten organic acids and ten combinations was determined. The testing of minimum inhibitory concentration (MIC) values was performed at pH 6.0 and 7.3 by using the broth microdilution method and included the following organic acids: Caprylic acid, sorbic acid, caproic acid, benzoic acid, ascorbic acid, propionic acid, acetic acid, formic acid, fumaric acid and tartaric acid and combinations thereof. The lowest MIC values were seen for caprylic acid (MIC range at pH 7.3: 0.5-2 mmol/L) and sorbic acid (MIC range at pH 7.3: 1-4 mmol/L). One to two dilution steps lower MIC values were determined at the lower pH value of 6.0. Furthermore, ten combinations consisting of three to five organic acids were developed. In addition to the tested antibacterial activity, other criteria were included such as approval as feed additives, reported synergistic effects and chemical properties. For nine of ten combinations, the MIC90 values of the organic acids decreased 1.25- to 241.5-fold compared to the MIC90 values for the individual substances. Furthermore, nine of ten combinations exhibited synergistic activities against two or more of the tested C. jejuni and C. coli isolates. A combination of caprylic acid, sorbic acid and caproic acid exhibited synergistic activities against the largest number of Campylobacter spp. isolates (six C. jejuni and four C. coli) with fractional inhibitory concentration (FIC) indices (∑FIC) ranging from 0.33 to 1.42. This study shows in vitro synergistic activities of different organic acids in combinations against the major Campylobacter species and could therefore be a promising basis for reducing Campylobacter spp. in vivo.

Effects of a chemical additive on the fermentation, microbial communities, and aerobic stability of corn silage with or without air stress during storage.

We evaluated the effects of a chemical additive on the microbial communities, fermentation profile, and aerobic stability of whole-plant corn silage with or without air stress during storage. Whole-plant corn was either untreated or treated with a chemical additive containing sodium benzoate, potassium sorbate, and sodium nitrite at 2 or 3 liters/t of fresh forage weight. Ten individually treated and replicated silos (7.5 liters) were made for each treatment. Half of the silos remained sealed throughout a 63-d storage period, and the other half was subjected to air stress for 2 h/wk. The composition of the bacterial and fungal communities of fresh forage and silages untreated or treated with 2 liters/t of fresh forage weight was analyzed by Illumina Miseq sequencing. Treated silage had greater (P < 0.05) aerobic stability than untreated, even when subjected to air stress during storage, but the numbers of yeasts culturable on selective agar were not affected. However, the additive reduced the relative abundance (RA) of the lactating-assimilating yeast Candida tropicalis (P < 0.01). In air-stressed silages, untreated silage had a greater (P < 0.05) RA of Pichia kudriavzevii (also a lactate assimilator) than treated silage, whereas treated silage was dominated by Candida humilis, which is usually unable to assimilate lactate or assimilates it slowly. The additive improved the aerobic stability by specifically preventing the dominance of yeast species that can consume lactate and initiate aerobic spoilage. To the best of our knowledge, this is the first work that identifies the specific action of this additive on shifting the microbial communities in corn silage.

Antibacterial Effect of Caprylic Acid and Potassium Sorbate in Combination against Listeria monocytogenes ATCC 7644.

ABSTRACT: Listeria monocytogenes is a common foodborne pathogen that cause life-threatening infection with high mortality rates. Biofilm development of L. monocytogenes decreases its sensitivity to antibiotics, which has long attracted attention globally. Caprylic acid (CA) and potassium sorbate (PS) are both widely used food preservatives, but their synergistic effect against L. monocytogenes has not been described. This study explored the antibacterial activities of the CA-PS combination against L. monocytogenes ATCC 7644 grown in planktonic or biofilm cultures. The fractional inhibitory concentration index values, determined by the checkerboard microdilution method, were 0.37 ± 0.03 and 0.31 ± 0.04, showing their synergistic antimicrobial effects against L. monocytogenes ATCC 7644 in planktonic and biofilm cultures, respectively. CA-PS effectively eradicated the biofilm biomass to 10.8% by crystal violet assay and to 8.63% by fluorescence microscopic analysis compared with the control. The apoptosis rates of microbial cells embedded within biofilm significantly increased to 51.4%. Subsequent analysis revealed that the combination inhibited biofilm formation by affecting extracellular DNA release and polysaccharide intercellular adhesion expression, which was decreased from 8.93 to 1.04 ng of extracellular DNA per relative biomass and to 54.7% of the control, respectively. In addition, the combination inhibited the growth of L. monocytogenes ATCC 7644 by up to 0.67 ± 0.05 and 0.30 ± 0.03 log CFU/cm2 in planktonic and biofilm modes on a carrot surface, respectively. The synergistic antibacterial effects of CA-PS against L. monocytogenes ATCC 7644 were statistically significant, and the combination is an excellent candidate to be a novel food preservative.