Antilisterial Activity Research Articles

Antilisterial effect of alkyl polyglycosides biosurfactant and modes of action

Several outbreaks of Listeria monocytogenes originated mainly from contaminated contact surfaces. Thus, this study investigated the antilisterial effect of natural surfactants in terms of their use as a 2-in-1 sanitizing washer on a food contact surface and evaluated their modes of action. The antilisterial activity of alkyl polyglycosides (APGs), namely capryl glucoside (CA), coco glucoside (CG), and decyl glucoside (DG), was evaluated based on the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) using broth dilution assay. The results showed that CG had the strongest antilisterial activity. Therefore, CG was selected for further investigation. The time-kill assay showed a lethal effect of 0.5 % (w/w) CG by inactivating 4 Log reduction (99.99 %) of L. monocytogenes within 3 s. Furthermore, 1 % (w/w) CG with slight mechanical force in washing (by shaking) was efficient for sanitizing a stainless-steel coupon surface based on its ability to cause a total reduction of deposited L. monocytogenes (99.9 %) within 10 min. Scanning electron microscopy and applying Fourier-transform infrared spectroscopy revealed that CG chemically disrupted the cell wall and plasma membrane of L. monocytogenes within 5 min after a gentle wash. The results showed it had potent antimicrobial activity and was bactericidal against L. monocytogenes. Overall, our results supported the use of CG as a natural antibacterial surfactant to alter the chemical sanitizer and the possibility of its practical use in the food industry and for household use.

Bacteriocin characterization of Enterococcus faecium isolates and evaluation of their in situ anti-Listerial activity in Beyaz cheese

Nowadays, as the interest in natural preservatives increases, research on bioprotective cultures that produce bacteriocins and studies on the use of these cultures in foods are also increasing day by day. Present study aimed to investigate bacteriocin characterization and safety of Enterococcus faecium H108, H206 isolates and their potential utilization in the reduction of Listeria monocytogenes ATCC7644 in Beyaz cheese. Enterocin producing E. faecium H108 and H206 strains were isolated from raw cow's milk and identified by 16S rRNA sequencing. Enterocin containing supernatants of E. faecium H108 and H206 were found to be stable to heat various chemicals used in food industry, wide pH range, lysozyme and catalase enzymes. On the other hand, activity of enterocins were inactivated by proteolytic enzymes. Maximum activity of E. faecium H108 was 6400 AU/mL, while that of H206 was 12800 AU/mL, and the activities remained stable for 24 h. Supernatant of both E. faecium H108 and H206 inhibited L. monocytogenes growth in vitro. E. faecium H108 and H206 did not show haemolytic and DN-ase activities and were susceptible to most of the antibiotics tested. The isolates were used in Beyaz cheese and were found to significantly, but not completely inhibit L. monocytogenes growth. The findings of this research suggest that E. faecium H108 and H206 isolates, after a detailed safety assessment, might be employed as bioprotective cultures in Beyaz cheese with starter culture or hurdle technologies.

Single and combined application of bacteriophage and cinnamon oils against pathogenic Listeria monocytogenes in milk and smoked salmon

Nowadays, the discovery of alternative natural antimicrobial substances such as bacteriophages, essential oils, and other physical and chemical agents is developing in the food industry. In this study, nine bacteriophages were isolated from various parts of raw chickens and exhibited lytic activities against L. monocytogenes and various Listeria spp. The characterization of phage vB_LmoS-PLM9 was stable at 4 to 50 °C and pH range from 4 to 10. Phage vB_LmoS-PLM9 had a circular, double-stranded genomic DNA with 38,345 bp having endolysin but no antibiotic resistance or virulence genes. Among the eight essential oils tested at 10 %, cinnamon bark, and cassia oils showed the strongest antilisterial activities. The combined use of phage vB_LmoS-PLM9 and cinnamon oils indicated higher efficiency than single treatments. The combination of phage (MOI of 10) and both cinnamon oils (0.03 %) reduced the viable counts of L. monocytogenes and inhibited the regrowth of resistant cell populations in broth at 30 °C. Furthermore, treatment with the combination of phage (MOI of 100) and cinnamon oil (0.125 %) was effective in milk, especially at 4 °C by reducing the viable count to less than lower limit of detection. These results suggest combining phage and cinnamon oil is a potential approach for controlling L. monocytogenes in milk.

Production of bacterial cellulose-based peptidopolysaccharide BC-L with anti-listerial properties using a co-cultivation strategy

Bacterial cellulose (BC) has been found extensive applications in diverse domains for its exceptional attributes. However, the lack of antibacterial properties hampers its utilization in food and biomedical sectors. Leucocin, a bacteriocin belonging to class IIa, is synthesized by Leuconostoc that demonstrates potent efficacy against the foodborne pathogen, Listeria monocytogenes. In the current study, co-culturing strategy involving Kosakonia oryzendophytica FY-07 and Leuconostoc carnosum 4010 was used to confer anti-listerial activity to BC, which resulted in the generation of leucocin-containing BC (BC-L). The physical characteristics of BC-L, as determined by X-ray diffraction (XRD), Fourier-transform infrared spectroscopy (FTIR), and thermogravimetric analysis (TGA), were similar to the physical characteristics of BC. Notably, the experimental results of disc diffusion and growth curve indicated that the BC-L film exhibited a potent inhibitory effect against L. monocytogenes. Scanning electron microscopy (SEM) showed that BC-L exerts its bactericidal activity by forming pores on the bacterial cell wall. Despite the BC-L antibacterial mechanism, which involves pore formation, the mammalian cell viability remained unaffected by the BC-L film. The measurement results of zeta potential indicated that the properties of BC changed after being loaded with leucocin. Based on these findings, the anti-listerial BC-L generated through this co-culture system holds promise as a novel effective antimicrobial agent for applications in meat product preservation and packaging.

Antilisterial activity of olive-derived polyphenols: an experimental study on meat preparations

Pork meat and processed pork products have been linked to multiple listeriosis outbreaks worldwide during the past years. Specifically, it has been highlighted that minced pork meat is easily perishable and may increase the growth of Listeria monocytogenes, which could be harmful to the general public's health. This study aimed to investigate the potential application of olive oil mill wastewater polyphenolic and red beet extracts as natural antimicrobial agents for L. monocytogenes growth control in burgers. The minced pork meat was mixed with the extracts and experimentally inoculated with L. monocytogenes, then molded into vacuum-packaged and cold-stored (4±1°C) burgers kept under alternating exposure to fluorescent light. The L. monocytogenes enumeration was performed on burgers at 0, 2, 5, and 10 days of shelf life. In uninoculated burgers, physicochemical (pH, water activity, color) and sensory determination (descriptive sensory analysis) were also conducted. At the end of storage, the samples treated with olive-derived extract showed the lowest value of L. monocytogenes (approximately 1.3 Log CFU/g). The physicochemical and sensory traits of burgers have benefited from the addition of both olive-derived and red beet extracts. Results suggest that olive mill wastewater polyphenolic extracts could be added to minced pork meat products to act as a natural antimicrobial agent.

Inactivation of Listeria monocytogenes on a laboratory medium and enoki mushrooms using organic acid and essential oil vapors

The aim of this study was to determine the antimicrobial activity of organic acid (OA) and essential oil (EO) vapors against Listeria monocytogenes on a laboratory medium and enoki mushrooms. Among 14 OA and 8 EO vapors tested, 3 OA vapors (acetic, formic, and propionic acid) and 3 EO vapors (cinnamon bark, citronella, and lemongrass) showed relatively strong antilisterial activity. On tryptic soy agar (TSA), all three OA vapors had a minimum inhibitory concentration (MIC) of 12.5 mg/L, while the minimum lethal concentration (MLC) of formic acid vapor was the lowest, at 50 mg/L, followed by acetic acid vapor (200 mg/L) and propionic acid vapor (400 mg/L). The MIC values of lemongrass, cinnamon bark, and citronella EO vapors were 25, 100, and 100 mg/L, respectively. The MLC of lemongrass EO vapor was 100 mg/L and MLCs of other EO vapors were >400 mg/L. Formic acid and lemongrass EO vapors showed a partial synergism on TSA (fractional inhibitory concentration index of 0.7500). For L. monocytogenes inoculated on enoki mushrooms, formic acid vapor at a concentration of ≥50 mg/L, as well as formic acid and lemongrass vapors in combination (1:2 vol ratio) at ≥ 75 mg/L, inactivated L. monocytogenes. These results indicate that formic acid vapor is effective at controlling the growth of L. monocytogenes on enoki mushrooms.

A Mini-Review of Anti-Listerial Compounds from Marine Actinobacteria (1990-2023).

Among the foodborne illnesses, listeriosis has the third highest case mortality rate (20-30% or higher). Emerging drug-resistant strains of Listeria monocytogenes, a causative bacterium of listeriosis, exacerbate the seriousness of this public health concern. Novel anti-Listerial compounds are therefore needed to combat this challenge. In recent years, marine actinobacteria have come to be regarded as a promising source of novel antimicrobials. Hence, our aim was to provide a narrative of the available literature and discuss trends regarding bioprospecting marine actinobacteria for new anti-Listerial compounds. Four databases were searched for the review: Academic Search Ultimate, Google Scholar, ScienceDirect, and South African Thesis and Dissertations. The search was restricted to peer-reviewed full-text manuscripts that discussed marine actinobacteria as a source of antimicrobials and were written in English from 1990 to December 2023. In total, for the past three decades (1990-December 2023), only 23 compounds from marine actinobacteria have been tested for their anti-Listerial potential. Out of the 23 reported compounds, only 2-allyoxyphenol, adipostatins E-G, 4-bromophenol, and ansamycins (seco-geldanamycin B, 4.5-dihydro-17-O-demethylgeldanamycin, and seco-geldanamycin) have been found to possess anti-Listerial activity. Thus, our literature survey reveals the scarcity of published assays testing the anti-Listerial capacity of bioactive compounds sourced from marine actinobacteria during this period.

Attenuation of biofilm formation of Listeria monocytogenes by bacteriocin producing Pediococcus pentosaceus DS1 on selected biotic and abiotic surfaces

The present study explores the antilisterial activities of potentially probiotic strain Pediococcus pentosaceus DS1 isolated from fermented bamboo shoot. 16s rRNA gene sequence analysis confirmed the identity of the strains. Class IIA bacteriocin pediocin expressing YGNGV motif was identified and its antilisterial activity was validated by well diffusion assay. When cocultured with P. pentosaceus DS1, in case of L. monocytogenes AMDK2, maximum biofilm exclusion (55.54 ± 3.44%) was observed which could be visually observed by Scanning electron micrographs. P. pentosaceus DS1 showed maximum adhesion inhibition to Caco-2 cell line in case of L. monocytogenes AMDK2 (91.8%) whereas maximum decrease in invasion was observed in case of L. monocytogenes MTCC 839 (52.9%). Coculture in UHT milk also significantly decreased microbial growth parameters of Listeria suggesting the efficiency of P. pentosaceus DS1 in the removal of Listeria contamination from food.

Anti-Listerial Activity of Bacteriocin-like Inhibitory Substance Produced by Enterococcus lactis LBM BT2 Using Alternative Medium with Sugarcane Molasses.

Listeria monocytogenes is a foodborne pathogen that contaminates food-processing environments and persists within biofilms on equipment, thus reaching final products by cross-contamination. With the growing demand for clean-label products, the search for natural antimicrobials as biopreservants, such as bacteriocins, has shown promising potential. In this context, this study aimed to evaluate the anti-listerial action of bacteriocins produced by Enterococcus lactis LBM BT2 in an alternative medium containing sugarcane molasses (SCM). Molecular analyses were carried out to characterize the strain, including the presence of bacteriocin-related genes. In the kinetic study on SCM medium E. lactis, LBM BT2 showed biomass and bacteriocin productions similar to those observed on a sucrose-based medium (control), highlighting the potential of the sugarcane molasses as a low-cost substrate. Stability tests revealed that the molecule remained active in wide ranges of pH (4-10) and temperature (60-100 °C). Furthermore, the proteolytic treatment reduced the biomolecule's antimicrobial activity, highlighting its proteinaceous nature. After primary purification by salting out and tangential flow filtration, the bacteriocin-like inhibitory substance (BLIS) showed bacteriostatic activity on suspended L. monocytogenes cells and against biofilm formation at a concentration of 0.625 mg/mL. These results demonstrate the potential of the produced BLIS as a biopreservative in the food industry.

Enterocins Produced by Enterococci Isolated from Breast-Fed Infants: Antilisterial Potential.

Enterocins are bacteriocins synthesized by Enterococcus strains that show an interesting antimicrobial effectiveness against foodborne pathogens such as Listeria monocytogenes. The objectives of this study were to identify and analyze the expression of enterocin genes of Enterococcus isolated from breast-fed infants and evaluate their ability to inhibit three human isolates of virulent L. monocytogenes, as well as some probiotic bacteria. The susceptibility of the strains of L. monocytogenes to fifteen antibiotics was tested, detecting their resistance to cefoxitin (constitutively resistant), oxacillin, and clindamycin. The production of enterocins A, B, and P was observed in Enterococcus faecium isolates, while enterocin AS-48 was detected in an Enterococcus faecalis isolate. AS-48 showed antilisterial activity by itself, while the joint action of enterocins A and B or B and P was necessary for inhibiting L. monocytogenes, demonstrating the synergistic effect of those combinations. The presence of multiple enterocin genes does not assure the inhibition of L. monocytogenes strains. However, the expression of multiple enterocin genes showed a good correlation with the inhibition capacity of these strains. Furthermore, the potential beneficial strains of lactobacilli and bifidobacteria examined were not inhibited by any of the enterocins produced individually or in combination, with the exception of Bifidobacterium longum BB536, which was inhibited by enterocin AS-48 and the joint production of enterocins A and B or B and P. The enterocins studied here could be candidates for developing alternative treatments against antibiotic-resistant bacterial infections. Moreover, these selected enterocin-producing E. faecium strains isolated from breast-fed infants could be used as probiotic strains due to their antilisterial effect, as well as the absence of virulence factors.

Isolation and characterization of lactic acid bacteria producing a potent anti-listerial bacteriocin-like inhibitory substance (BLIS) from chhurpi, a fermented milk product.

Nowadays, the bacteriocin industries have seen significant growth, supplanting chemical preservatives in its ability to improve the shelf-life and safety of food. The increasing customer desire to use natural preservatives has fueled advancing bacteriocin research. The objective of this study was to identify lactic acid bacteria (LAB) that produce bacteriocin-like inhibitory substance (BLIS) and have strong anti-listerial activity. We have identified and analyzed a LAB obtained from chhurpi samples, a popular milk-derived product in the Himalayan regions of India and Nepal. The strain was studied and identified based on its morphological, biochemical, and physiological characteristics. Furthermore, the molecular 16s-rDNA analysis suggests that the strain was Lactococcus sp. RGUAM1 (98.2% similar to Lactococcus lactis subsp. hordniae NBRC 100931T). The isolated strain can produce a potent BLIS, which has shown efficacy against three gram-positive bacteria responsible for food spoilage, such as Listeria monocytogenes (MTCC 657), Staphylococcus aureus subsp. aureus (MTCC 87), Lactobacillus plantarum (MTCC 1407), Lactobacillus paraplantarum (MTCC 12904). The scanning electron microscope (SEM) image illustrates that the crude cell-free supernatant (CFS) disrupts the cell envelope, leading to the release of cellular contents and the clustering of cells. In addition, this BLIS can easily withstand a wide range of pH (2-12), temperature (up to 100°C for 15min), bile salt (0.3% W/V), salinity (4% W/V), and enzyme activity of 1600AU/ml against Listeria monocytogenes. Our research offers a robust framework and valuable insights into bio-preservation and its potential applications in diverse food products.

Thymbra capitata L. hydrolate as a washing solution for controlling Listeria monocytogenes and spoilage microorganisms on rocket salad

Ready-to-eat (RTE) vegetables, including fresh-cut ones, are exposed to the risk of the foodborne pathogen Listeria monocytogenes. Nowadays, fresh-cut products are widely consumed because of their convenience. Nevertheless, effective washing treatments are required for their preparation, to maintain freshness and microbiological safety. Currently, natural substances are increasingly demanded as alternatives to traditional synthetic washing agents. Therefore, this study investigates the application of Thymbra capitata hydrolate as washing solution for rocket salad (Eruca sativa) stored at 4 °C for 48 h, to evaluate a potential application both during the production process of fresh-cut vegetables, and also at the domestic level. The hydrolate applied at a concentration of 500 μL/mL for 5 min allowed to reduce L. monocytogenes load of about 1 log CFU/g at each time of analysis, and to determine a significant reduction of microbial groups normally found on rocket salad, particularly total mesophilic and psychrotrophic counts. The treatment showed no negative effects on pH, aw, and colour of the rocket salad. At a sensory level, the differences among control and treated samples were minimally perceived only on the first day of analysis but disappeared over time. Therefore, this alternative washing treatment revealed to counteract L. monocytogenes growth and to help maintaining the quality of rocket salad. Hence, it could be optimized for a concrete application, also considering the enhancement of the antilisterial activity, the sustainability and low cost of the hydrolate, which represents a by-product from the production of essential oils.

Antilisterial activity of raw sheep milk from two native Epirus breeds: Culture-dependent identification, bacteriocin gene detection and primary safety evaluation of the antagonistic LAB biota

Raw milk from native small ruminant breeds in Epirus, Greece, is a valuable natural source of autochthonous lactic acid bacteria (LAB) strains with superior biotechnological properties. In this study, two bulk milks (RM1, RM2) from two local sheep yards, intended for traditional Kefalotyri cheese production, were preselected for bacteriocin-like antilisterial activity by in vitro tests. Their antagonistic LAB biota was quantified followed by polyphasic (16S rRNA gene sequencing; IGS for Enterococcus; a multiplex-PCR for Leuconostoc) identification of 42 LAB (RM1/18; RM2/24) isolates further evaluated for bacteriocin encoding genes and primary safety traits. Representative isolates of the numerically dominant mesophilic LAB were Leuconostoc mesenteroides (10) in both RMs, Streptococcus parauberis (7) in RM2, and Lactococcus lactis (1) in RM1; the subdominant thermophilic LAB isolates were Enterococcus durans (8), E. faecium (6), E. faecalis (3), E. hirae (1), E. hermanniensis (1), Streptococcus lutetiensis (2), S. equinus (1) and S. gallolyticus (1). Based on their rpoB, araA, dsr and sorA profiles, six Ln. mesenteroides strains (8 isolates) were atypical lying between the subspecies mesenteroides and dextranicum, whereas two strains profiled with Ln. mesenteroides subsp. jonggajibkimchii that is first-time reported in Greek dairy food. Two RM1 E. faecium strain biotypes (3 isolates) showed strong, enterocin-mediated antilisterial activity due to entA/entB/entP possession. One E. durans from RM1 possessed entA and entP, while additional nine RM2 isolates of the E. faecium/durans group processed entA or entP singly. All showed direct (cell-associated) antilisterial activity only, as also both S. lutetiensis strains from RM2 did strongly. Desirably, no LAB isolate was β-hemolyrtic, or cytolysin-positive, or possessed vanA, vanB for vancomycin resistance, or agg, espA, hyl, and IS16 virulence genes. However, all three E. faecalis from RM2 possessed gelE and/or ace virulence genes. In conclusion, all Ln. mesenteroides strains, the two safe, enterocin A-B-P-producing E. faecium strains, and the two antilisterial S. lutetiensis strains should be validated further as potential costarter or adjunct cultures in Kefalotyri cheese. The prevalence of α-hemolytic pyogenic streptococci in raw milk, mainly S. parauberis in RM2, requires consideration in respect to subclinical mastitis in sheep and the farm hygiene overall.

Modulation of the Gut Microbiota by the Plantaricin-Producing Lactiplantibacillus plantarum D13, Analysed in the DSS-Induced Colitis Mouse Model.

Lactiplantibacillus plantarum D13 shows antistaphylococcal and antilisterial activity, probably due to the synthesis of a presumptive bacteriocin with antibiofilm capacity released in the cell-free supernatant (CFS), whose inhibitory effect is enhanced by cocultivation with susceptible strains. An in silico analysis of the genome of strain D13 confirmed the pln gene cluster. Genes associated with plantaricin biosynthesis, structure, transport, antimicrobial activity, and immunity of strain D13 were identified. Furthermore, the predicted homology-based 3D structures of the cyclic conformation of PlnE, PlnF, PlnJ, and PlnK revealed that PlnE and PlnK contain two helices, while PlnF and PlnJ contain one and two helices, respectively. The potential of the strain to modulate the intestinal microbiota in healthy or dextran sulphate sodium (DSS)-induced colitis mouse models was also investigated. Strain D13 decreased the disease activity index (DAI) and altered the gut microbiota of mice with DSS-induced colitis by increasing the ratio of beneficial microbial species (Allobaculum, Barnesiella) and decreasing those associated with inflammatory bowel disease (Candidatus Saccharimonas). This suggests that strain D13 helps to restore the gut microbiota after DSS-induced colitis, indicating its potential for further investigation as a probiotic strain for the prevention and treatment of colitis.

A Novel Two-Component Bacteriocin, Acidicin P, and Its Key Residues for Inhibiting Listeria monocytogenes by Targeting the Cell Membrane.

Listeria monocytogenes is an important pathogen which easily contaminates food and causes fatal systemic infections in human. Bacteriocins have received much attention regarding their natural methods of controlling health-related pathogens. Here, we investigated and characterized a novel two-component bacteriocin named acidicin P from Pediococcus acidilactici LAC5-17. Acidicin P showed obvious antimicrobial activity to L. monocytogenes. Through a sequence similarity network analysis for two-component bacteriocin precursors mined in the RefSeq database, acidicin P was observed to belong to an unusual group of two-component bacteriocins. Acidicin P contains two peptides designated Adpα and Adpβ which are assessed to interact with each other and form a helical dimer structure which can be inserted into the lipid bilayer of target cell membrane. We demonstrate that A5, N7, and G9 in the A5xxxG9 motif of Adpα and S16, R19, and G20 in the S16xxxG20 motif of Adpβ played crucial roles in stabilizing the helix-helix interaction of Adpα and Adpβ and were essential for the antilisterial activity of acidicin P by site-directed mutagenesis. A positive residue, R14, in Adpα and a negative residue, D12, in Adpβ are also important for acidicin P to fight against L. monocytogenes. These key residues are supposed to form hydrogen bonding, which is crucial for the interaction of Adpα and Adpβ. Furthermore, acidicin P induces severe permeabilization and depolarization of the cytoplasmic membrane and causes dramatic changes in L. monocytogenes cell morphology and ultrastructure. Acidicin P has the potential to be applied to inhibit L. monocytogenes efficiently both in the food industry and medical treatments. IMPORTANCE L. monocytogenes can cause widespread food contamination and severe human listeriosis, which amount to a large proportion of the public health and economic burdens. Today, L. monocytogenes is usually treated with chemical compounds in the food industry or antibiotics for human listeriosis. Natural and safe antilisterial agents are urgently required. Bacteriocins are natural antimicrobial peptides that have comparable narrow antimicrobial spectra and are attractive potentials for precision therapy for pathogen infection. In this work, we discover a novel two-component bacteriocin designated acidicin P, which shows obvious antilisterial activity. We also identify the key residues in both peptides of acidicin P and demonstrate that acidicin P is inserted into the target cell membrane and disrupts the cell envelop to inhibit the growth of L. monocytogenes. We believe that acidicin P is a promising lead for further development as an antilisterial drug.

Bacteriocin-like inhibitory substances as green bio-preservatives; nanoliposomal encapsulation and evaluation of their in vitro/in situ anti-Listerial activity

In this study, predominant lactic acid bacteria (LAB) isolated from a traditional pickle (Persian pickled Litteh) were screened based on their ability to produce bacteriocin-like inhibitory substances (BLIS) with inhibitory effects on foodborne bacteria tested. The isolated BLIS was also characterized using liquid chromatography-mass spectrometry (LC-MS), and loaded into nanoliposomes; then, it's in vitro and in situ inhibitory effects were evaluated. Sequencing results of the PCR products led to the identification of Lactococcus lactis LI04 as the selected BLIS-producing LAB. The highest inhibitory activity of the BLIS was observed against Listeria monocytogenes PTCC 1298, which was significantly (P < 0.05) higher than the other foodborne bacteria studied. According to LC-MS analysis, presence of two peptides with retention times of 1.45 and 1.76 min corresponding to mass to charge ratio of 456.45 and 194.79 was approved in the BLIS. Produced empty and BLIS-loaded nanoliposomes had diameters of 92.99 and 139.10 nm, respectively. Results of Fourier transform infrared analysis showed that BLIS loading induced high degree of hydration in nanoliposomes. The increased Tm of loaded nanoliposomes (verified by differential scanning calorimetry) implied that the rigidity of liposomal membrane was enhanced. The loaded nanoliposomes were also able to maintain their inhibitory activity after treatment in simulated gastrointestinal conditions and commercial sterilization; whereas, the free (non-encapsulated) BLIS lost its effect. Furthermore, anti-Listerial activity of nanoliposomal BLIS after pasteurization was higher than the free BLIS. The highest release of the BLIS from nanoliposomes at 4 °C was observed after 16 days' storage. After this period, anti-Listerial activity of nanoliposomal BLIS was significantly higher than the free BLIS in the produced homemade beef steak.

Selection of lactic acid bacteria as biopreservation agents and optimization of their mode of application for the control of Listeria monocytogenes in ready-to-eat cooked meat products

In order to meet consumers´ demands for more natural foods and to find new methods to control foodborne pathogens in them, research is currently being focused on alternative preservation approaches, such as biopreservation with lactic acid bacteria (LAB). Here, a collection of lactic acid bacteria (LAB) isolates was characterized to identify potential biopreservative agents. Six isolates (one Lactococcus lactis, one Lacticaseibacillus paracasei and four Lactiplantibacillus plantarum) were selected based on their antimicrobial activity in in vitro assays. Whole genome sequencing showed that none of the six LAB isolates carried known virulence factors or acquired antimicrobial resistance genes, and that the L. lactis isolate was potentially a nisin Z producer. Growth of L. monocytogenes was successfully limited by L. lactis ULE383, L. paracasei ULE721 and L. plantarum ULE1599 throughout the shelf-life of cooked ham, meatloaf and roasted pork shoulder. These LAB isolates were also applied individually or as a cocktail at different inoculum concentrations (4, 6 and 8 log10 CFU/g) in challenge test studies involving cooked ham, showing a stronger anti-Listerial activity when a cocktail was used at 8 log10 CFU/g. Thus, a reduction of up to ~5.0 log10 CFU/g in L. monocytogenes growth potential was attained in cooked ham packaged under vacuum, modified atmosphere packaging or vacuum followed by high pressure processing (HPP). Only minor changes in color and texture were induced, although there was a significant acidification of the product when the LAB cultures were applied. Remarkably, this acidification was delayed when HPP was applied to the LAB inoculated batches. Metataxonomic analyses showed that the LAB cocktail was able to grow in the cooked ham and outcompete the indigenous microbiota, including spoilage microorganisms such as Brochothrix. Moreover, none of the batches were considered unacceptable in a sensory evaluation.Overall, this study shows the favourable antilisterial activity of the cocktail of LAB employed, with the combination of HPP and LAB achieving a complete inhibition of the pathogen with no detrimental effects in physico-chemical or sensorial evaluations, highlighting the usefulness of biopreservation approaches involving LAB for enhancing the safety of cooked meat products.

Antilisterial activity of cinnamon essential oil, pomegranate extract, or strawberry tree extract against Listeria monocytogenes in slices of dry-cured ham and pork loin.

Owing to concerns about the antimicrobial resistance of agents that can prevent the growth of Listeria monocytogenes in meat, researchers have investigated natural preservatives with antilisterial effects. However, in vivo application of essential oils and plant extracts usually results in reduced antimicrobial activity in meat products when compared to in vitro studies. This study aimed to evaluate the in vivo antimicrobial activity of cinnamon essential oil, pomegranate, and strawberry tree extracts in slices of dry-cured ham and pork loin against L. monocytogenes. Fragments of sterile dry-cured ham were inoculated with 100 μL cinnamon oil 0.5%, pomegranate, or strawberry crude extract. After 10 min, 100 μL of L. monocytogenes serotype 4b (104 colony-forming unit [CFU]/mL) was inoculated, and samples were incubated at 7 °C for 7 d to simulate the processing and storage temperature conditions of dry-cured meat products. L. monocytogenes was detected and quantified. Only strawberry extract presented significant differences (P < 0.05) from the control; thus, it was selected for the assay with 2% and 4% salt-treated pork loin. The strawberry tree extract significantly (P < 0.05) reduced the growth of L. monocytogenes in dry-cured ham. However, it could not reduce L. monocytogenes growth in pork loin, regardless of the salt concentration. This is the first report on the antimicrobial effect of strawberry tree leaf extract against L. monocytogenes in dry-cured ham.

Anti-Listerial Activity Control of Postbiotic in Cow Milk Lump Cheese Processing

Anti-Listerial Activity Control of Postbiotic in Cow Milk Lump Cheese Processing

Potential of Essential Oils in the Control of Listeria monocytogenes.

Listeria monocytogenes is a foodborne pathogen, the causative agent of listeriosis. Infections typically occur through consumption of foods, such as meats, fisheries, milk, vegetables, and fruits. Today, chemical preservatives are used in foods; however, due to their effects on human health, attention is increasingly turning to natural decontamination practices. One option is the application of essential oils (EOs) with antibacterial features, since EOs are considered by many authorities as being safe. In this review, we aimed to summarize the results of recent research focusing on EOs with antilisterial activity. We review different methods via which the antilisterial effect and the antimicrobial mode of action of EOs or their compounds can be investigated. In the second part of the review, results of those studies from the last 10 years are summarized, in which EOs with antilisterial effects were applied in and on different food matrices. This section only included those studies in which EOs or their pure compounds were tested alone, without combining them with any additional physical or chemical procedure or additive. Tests were performed at different temperatures and, in certain cases, by applying different coating materials. Although certain coatings can enhance the antilisterial effect of an EO, the most effective way is to mix the EO into the food matrix. In conclusion, the application of EOs is justified in the food industry as food preservatives and could help to eliminate this zoonotic bacterium from the food chain.