Antioxidant and antimicrobial extracts from grape stalks obtained with subcritical water. Potential use in active food packaging development.

In order to valorise winemaking grape stalks, subcritical water extraction at 160 and 180 °C has been carried out to obtain phenolic-rich extracts useful for developing active food packaging materials. Red (R) and white (W) varieties (from Requena, Spain) were used, and thus, four kinds of extracts were obtained. These were characterised as to their composition, thermal stability and antioxidant and antibacterial activity. The extracts were incorporated at 6 wt% into polylactic acid (PLA) films and their effect on the optical and barrier properties of the films and their protective effect against sunflower oil oxidation was analysed. Carbohydrates were the major compounds (25-38%) in the extracts that contained 3.5-6.6% of phenolic compounds, the R extracts being the richest, with higher radical scavenging capacity. Every extract exhibited antibacterial effect against Escherichia coli and Listeria innocua, while PLA films with extracts preserved sunflower oil against oxidation.

Antimicrobial activity of Tetradenia riparialeaf essential oil / Actividad antimicrobiana del aceite esencial de hoja de Tetradenia riparia

Food spoilage is a widely neglected problem and the constant use of synthetic fungicides could develop resistant fungi. The objective of this study was to evaluate the chemical composition and antimicrobial activity of Tetradenia riparialeaf essential oil against foodborne disease microorganisms. Leaf essential oil was obtained by hydrodistillation and identified by gas chromatography coupled to mass spectrometry. The antimicrobial activity was studied by broth microdilution. The major compounds identified were oxygenated sesquiterpenes (43.6%): 14-hydroxy-9-epi-(E)-cariophylene (20.8%) and τ-cadinol (18.4%); followed by oxygenated diterpenes (24.6%): 6,7-dehydroroyleanone (12.6%) and 9ß, 13ß-epoxy-7-abiethene (10.6%); sesquiterpenic hydrocarbons (17.1%) and oxygenated monoterpenes (7.4%): fenchone (5.6%). The essential oil had broad antibacterial and antifungal activity, mainly against A. versicolor and P. ochrochloron with fungistatic and fungicidal activities and B. cereus, L. monocytogenes, and S. aureuswith bacteriostatic and bactericidal activities. T. riparialeaf essential oil is a potential alternative to control microorganisms-

El deterioro de los alimentos es un problema ampliamente desatendido y el uso constante de fungicidas sintéticos podría desarrollar hongos resistentes. El objetivo de este estudio fue evaluar la composición química y la actividad antimicrobiana del aceite esencial de hoja de Tetradenia riparia contra microorganismos patógenos transmitidos por los alimentos. El aceite esencial de hoja se obtuvo por hidrodestilación y se identificó mediante cromatografía de gases acoplada a espectrometría de masas. La actividad antimicrobiana estudiada fue por microdilución en caldo. Los compuestos principales del aceite esencial se identificaron como sesquiterpenos oxigenados (43,6%): 14-hidroxi-9-epi-(E)-cariofileno (20,8%) y τ-cadinol (18,4%); seguido de diterpenos oxigenados (24,6%): 6-7-deshidroroileanona (12,6%) y 9ß, 13ß-epoxi-7-abieteno (10,6%); hidrocarburos sesquiterpénicos (17,1%) y monoterpenos oxigenados (7,4%): fenchona (5,6%). Tenía amplia actividad antibacteriana y antifúngica, principalmente contra A. versicolor y P. ochrochloron con actividades fungistáticas y fungicidas, y principalmente contra B. cereus, L. monocytogenes y S. aureus con actividades bacteriostáticas y bactericidas. El aceite esencial de hoja de T. riparia es una alternativa potencial para controlar microorganismos.

Proteomic Characterization of Antibiotic Resistance in Listeria and Production of Antimicrobial and Virulence Factors.

Some Listeria species are important human and animal pathogens that can be found in contaminated food and produce a variety of virulence factors involved in their pathogenicity. Listeria strains exhibiting multidrug resistance are known to be progressively increasing and that is why continuous monitoring is needed. Effective therapy against pathogenic Listeria requires identification of the bacterial strain involved, as well as determining its virulence factors, such as antibiotic resistance and sensitivity. The present study describes the use of liquid chromatography-electrospray ionization tandem mass spectrometry (LC-ESI-MS/MS) to do a global shotgun proteomics characterization for pathogenic Listeria species. This method allowed the identification of a total of 2990 non-redundant peptides, representing 2727 proteins. Furthermore, 395 of the peptides correspond to proteins that play a direct role in Listeria pathogenicity; they were identified as virulence factors, toxins and anti-toxins, or associated with either antibiotics (involved in antibiotic-related compounds production or resistance) or resistance to toxic substances. The proteomic repository obtained here can be the base for further research into pathogenic Listeria species and facilitate the development of novel therapeutics for these pathogens.

Influence of molecular weight fractionation on the antimicrobial and anticancer properties of a fucoidan rich-extract from the macroalgae Fucus vesiculosus.

The objective of this study was to investigate the antimicrobial and anticancer properties of a fucoidan extract and subsequent fractions isolated from the macroalgae Fucus vesiculosus. The fractions obtained (>300 kDa, <300 kDa, <100 kDa, <50 kDa and <10 kDa) could inhibit the growth of B. subtilis, E. coli, L. innocua and P. fluorescens when assayed at concentrations between 12,500 and 25,000 ppm. The bacterial growth was monitored by optical density (OD) measurements (600 nm, 24 h) at 30 °C or 37 °C, depending upon on the strain used. The extracted fractions were also tested for cytotoxicity against brain glioblastoma cancer cells using the Alamar Blue assay for 24 h, 48 h and 6 days. The >300 kDa fraction presented the lowest IC50 values (0.052% - 24 h; 0.032% - 6 days). The potential bioactivity of fucoidan as an antimicrobial and anticancer agent was demonstrated in this study. Hence, the related mechanisms of action should be explored in a near future.

Incorporation of Antimicrobial Bio-Based Carriers onto Poly(vinyl alcohol-co-ethylene) Surface for Enhanced Antimicrobial Activity.

A biobased rechargeable antimicrobial modification approach was developed using a covalent immobilization of food grade yeast cell wall particles on a model plastic film. We demonstrate the applications of this modification approach on poly(vinyl alcohol-co-ethylene) surface to inactivate inoculated bacteria with or without the presence of organic content, reducing the cross-contamination between food contact surface and model fresh produce, and inhibiting the growth of biofilms on the film surface. These biobased cell wall particle modified plastic films can enhance the binding of chlorine to the plastic surface in the form of N-halamine, extend the stability of chlorine against high organic content and ambient storage, and improve the rechargeability of the plastic films. Upon charging with chlorine, these modified plastic films inactivated 5 log of model Gram-negative bacteria (Escherichia coli O157:H7) and Gram-positive bacteria (Listeria innocua used as a surrogate of pathogenic Listeria monocytogenes) within 2 min of surface inoculation in water and within 20 min in an organic-rich aqueous environment. The modified plastic films prevented the transfer of bacteria and eliminated cross-contamination from the contaminated films to a spinach leaf surface, while 3 log CFU/leaf of bacteria were transferred from a contaminated native film to a noninoculated spinach surface. In addition, these modified plastic films reduced the adhesion of L. innocua cells by 2.7-3.6 log CFU/cm2 compared with control films during extended incubation for biofilm formation. Overall, this study demonstrates the feasibility of this biobased food grade modification approach to reduce microbial contamination and improve produce safety in the food processing industry.

Synergistic inactivation of bacteria based on a combination of low frequency, low-intensity ultrasound and a food grade antioxidant.

This study evaluated a synergistic antimicrobial treatment using a combination of low frequency and a low-intensity ultrasound (LFU) and a food-grade antioxidant, propyl gallate (PG), against a model gram-positive (Listeria innocua) and the gram-negative bacteria (Escherichia coli O157:H7). Bacterial inactivation kinetic measurements were complemented by characterization of biophysical changes in liposomes, changes in bacterial membrane permeability, morphological changes in bacterial cells, and intracellular oxidative stress upon treatment with PG, LFU, and a combination of PG + LFU. Combination of PG + LFU significantly (>4 log CFU/mL, P < 0.05) enhanced the inactivation of both L. innocua and E. coli O157:H7 compared to PG or LFU treatment. As expected, L. innocua had a significantly higher resistance to inactivation compared to E. coli using a combination of PG + LFU. Biophysical measurements in liposomes, bacterial permeability measurements, and scanning electron microscope (SEM)-based morphological measurements show rapid interactions of PG with membranes. Upon extended treatment of cells with PG + LFU, a significant increase in membrane damage was observed compared to PG or LFU alone. A lack of change in the intracellular thiol content following the combined treatment and limited effectiveness of exogenously added antioxidants in attenuating the synergistic antimicrobial action demonstrated that oxidative stress was not a leading mechanism responsible for the synergistic inactivation by PG + LFU. Overall, the study illustrates synergistic inactivation of bacteria using a combination of PG + LFU based on enhanced membrane damage and its potential for applications in the food and environmental systems.

Identification and Characterization of a Novel Genomic Island Harboring Cadmium and Arsenic Resistance Genes in Listeria welshimeri.

Listeria monocytogenes, the bacterial foodborne pathogen responsible for the severe disease listeriosis, frequently exhibits heavy metal resistance. Concurrent resistance to cadmium and arsenic in L. monocytogenes is strongly associated with the 35-kb chromosomal island LGI2. LGI2 has been encountered repeatedly among L. monocytogenes serotype 4b hypervirulent clones but, surprisingly, not among non-pathogenic Listeria spp. Here we describe a novel LGI2 variant, LGI2-3, in two L. welshimeri strains from an urban aquatic environment. Whole genome sequence analysis revealed that the genomes were closely related except for one prophage region and confirmed a chromosomally integrated LGI2-3. It harbored a cystathionine beta-lyase gene previously only encountered in LGI2-1 of L. monocytogenes clonal complex 1 but was otherwise most closely related to LGI2. LGI2-3 harbored a novel cadAC cassette (cadA7C7) that, like LGI2's cadA4C4, was associated with lower-level tolerance to cadmium (MIC 50 µg/mL) than other cadAC cassettes (MIC ≥ 140 µg/mL). CadA sequence analysis identified two amino acids that may be important for mediating different levels of cadmium tolerance. Our findings clearly demonstrated the potential for LGI2-like islands to be harbored by non-pathogenic Listeria spp. and generate intriguing hypotheses on the genetic diversity mediated by this island and its transfer among Listeria spp.

Combined Antimicrobial Effect of Bio-Waste Olive Leaf Extract and Remote Cold Atmospheric Plasma Effluent.

A novel strategy involving Olive Leaf Extract (OLE) and Cold Atmospheric Plasma (CAP) was developed as a green antimicrobial treatment. Specifically, we reported a preliminary investigation on the combined use of OLE + CAP against three pathogens, chosen to represent medical and food industries (i.e., E. coli, S. aureus and L. innocua). The results indicated that a concentration of 100 mg/mL (total polyphenols) in OLE can exert an antimicrobial activity, but still insufficient for a total bacterial inactivation. By using plain OLE, we significantly reduced the growth of Gram positive S. aureus and L. innocua, but not Gram-negative E. coli. Instead, we demonstrated a remarkable decontamination effect of OLE + CAP in E. coli, S. aureus and L. innocua samples after 6 h. This effect was optimally maintained up to 24 h in S. aureus strain. E. coli and L. innocua grew again in 24 h. In the latter strain, OLE alone was most effective to significantly reduce bacterial growth. By further adjusting the parameters of OLE + CAP technology, e.g., OLE amount and CAP exposure, it could be possible to prolong the initial powerful decontamination over a longer time. Since OLE derives from a bio-waste and CAP is a non-thermal technology based on ionized air, we propose OLE + CAP as a potential green platform for bacterial decontamination. As a combination, OLE and CAP can lead to better antimicrobial activity than individually and may replace or complement conventional thermal procedures in food and biomedical industries.

Sodium alginate-based edible coating containing nanoemulsion of Citrus sinensis essential oil eradicates planktonic and sessile cells of food-borne pathogens and increased quality attributes of tomatoes.

There is a growing interest from the worldwide scientific community in formulating edible- biodegradable coatings to replace non-biodegradable and expensive commercial wax-based coatings for preserving postharvest quality attributes of vegetables including tomatoes. Postharvest tomatoes are a suspected vehicle for both Salmonella and Listeria in food poisoning incidents. In this work, the effectiveness of edible nano-emulsion coatings containing sweet orange essential oil and sodium alginate were prepared and characterized, then evaluated antibacterial and antibiofilm activity against Salmonella and Listeria and simultaneously, examined its coating effect on various quality characteristics of tomatoes at 22 ± 2 °C over a 15 days storage period. DLS (Dynamic light scattering) study revealed stable nanoemulsion formulation with 43.23 nm particle size. The high whiteness index of nanoemulsion has a positive impact on product marketability and desirability. Antibacterial and antibiofilm studies revealed nanoemulsion effectively eradicate both sessile and planktonic forms of Salmonella and Listeria in both single and multi-species culture conditions. Tomatoes coated with edible coating significantly enhanced firmness up to 33%, decreased total mesophilic bacteria including Salmonella and Listeria, and reduced weight loss up to 3 fold lower than uncoated one. Sensory analysis revealed that the use of the edible coating increased the total acceptance scores of tomatoes.

Non-thermal atmospheric gas plasma for decontamination of sliced cheese and changes in quality.

The atmospheric-pressure non-thermal dielectric barrier discharge (DBD) plasma has recently emerged as an efficient decontamination method for the food safety enhancement. Thus the objective of this study was to evaluate the effectiveness of a simple DBD plasma treatment, with a relatively low-frequency power supply operating at 60 Hz, for microbial inactivation. A parametric study of operating conditions for bacterial inactivation was conducted using nutrient agar inoculated with Escherichia coli (2.28--6.28 log CFU/ml). The microbial log reduction was enhanced with increasing input power (30, 50, 70 W) and plasma exposure time (0, 1, 3, 5, 7 min). The inactivation effect was increased by decreasing inter-electrode gap (2, 1.5, 1 cm) and by reducing the initial microorganism concentration. Accordingly, a DBD plasma treatment at 50 W for 10 min could lead to complete killing of E. coli and partial inactivation of Listeria innocua on cheese (mean log reduction: 4.75 ± 0.02 and 0.72 ± 0.01, respectively). The decontamination efficacy of DBD plasma was affected by the types of microorganisms. The changes in hardness and color of cheese were unnoticeable after 10 min treatment with a power of 50 W. Overall, the results suggested that the DBD plasma can be potentially exploited to improve the food safety.

Oxysterols provide innate immunity to bacterial infection by mobilizing cell surface accessible cholesterol.

Cholesterol 25-hydroxylase (CH25H) is an interferon-stimulated gene that converts cholesterol to the oxysterol 25-hydroxycholesterol (25HC). Circulating 25HC modulates essential immunological processes including antiviral immunity, inflammasome activation and antibody class switching; and dysregulation of CH25H may contribute to chronic inflammatory disease and cancer. Although 25HC is a potent regulator of cholesterol storage, uptake, efflux and biosynthesis, how these metabolic activities reprogram the immunological state of target cells remains poorly understood. Here, we used recently designed toxin-based biosensors that discriminate between distinct pools of plasma membrane cholesterol to elucidate how 25HC prevents Listeria monocytogenes from traversing the plasma membrane of infected host cells. The 25HC-mediated activation of acyl-CoA:cholesterol acyltransferase (ACAT) triggered rapid internalization of a biochemically defined fraction of cholesterol, termed 'accessible' cholesterol, from the plasma membrane while having little effect on cholesterol in complexes with sphingomyelin. We show that evolutionarily distinct bacterial species, L. monocytogenes and Shigella flexneri, exploit the accessible pool of cholesterol for infection and that acute mobilization of this pool by oxysterols confers immunity to these pathogens. The significance of this signal-mediated membrane remodelling pathway probably extends beyond host defence systems, as several other biologically active oxysterols also mobilize accessible cholesterol through an ACAT-dependent mechanism.

Incorporation of natural antioxidants from rice straw into renewable starch films.

This study showed that rice straw waste is a valuable source for the extraction of water-soluble phenolic compounds that can be successfully incorporated into bioactive starch-based films. The major phenolic compounds in the extract were identified as ferulic, p-coumaric and protocatechuic acid using UHPLC-MS. Homogeneous films with antioxidant properties were produced by melt blending and compression molding and the changes in the physico-chemical properties were evaluated. The produced antioxidant starch films were slightly reddish-colored and exhibited good in-vitro antiradical scavenging activity against DPPH*. The addition of the antioxidant extract improved the oxygen barrier properties without negatively affecting the thermal and the water vapor barrier properties. However, antioxidant starch films turned more brittle with increasing amount of the antioxidant extract, which was probably due to interactions of phenolic compounds with the starch chains. The film forming process induced chain scission of starch molecules in all films, shown in a decrease in molecular weight of native starch from 9.1 × 106 Da to values as low as 1.0-3.5 × 106 Da. This study aids a circular economy by recycling rice straw for the production of bioactive food packaging.

Antioxidant and antimicrobial evaluation of rice bran (Oryza sativa L.) extracts in a mayonnaise-type emulsion.

The objective of this study was to evaluate the in vitro antimicrobial and antioxidant activities of different bran extracts and concentrations, and their influence on the parameters of a mayonnaise-type emulsion. To that end, first ethanol and then water were used to extract two rice bran extracts (RBE) from rice bran. Both these extracts were then added at two different concentrations (0.5 and 2%) to the emulsions that were subsequently analysed after seven days under two different storage temperatures, 4 °C and 20 °C. The antioxidant and antimicrobial ability of the extracts were evaluated, along with a control and a synthetic antioxidant. Results indicate the positive effect of rice bran extracts as additives in the food matrix. Ethanolic rice bran extract (EE) at 2% decreased the oxidation as well as mould and yeast proliferation and preserved the emulsion structure, while the other treatments acted in a similar way although their effect was less pronounced.

Cumin essential oil: Phytochemical analysis, antimicrobial activity and investigation of its mechanism of action through scanning electron microscopy.

In this study, the antimicrobial effects of cumin essential oil (CEO) and its mechanism of action through scanning electron microscopy (SEM) against Escherichia coli and Listeria innocua were investigated. The SEM images were taken at 0, 12 and 24 h at the minimum inhibitory concentration (MIC). The chemical composition of CEO was identified through gas chromatography/mass spectrometry (GC-MS). The antimicrobial effects of CEO were evaluated by the methods of Kirby-Bauer, well diffusion agar, microdilution broth and minimum bactericidal/fungicidal concentration (MBC/MFC). Antioxidant activity was examined by the methods of ß-carotene/linoleic acid inhibition and 2,2-diphenyl-1-picrylhydrazyl. Total phenol content (TPC) was measured by Folin-Ciocalteu method. The subsequent analysis of CEO through GC-MS revealed that cuminal (28.28%) was the major compound of CEO. CEO showed a high TPC of 89.45 ±â€¯0.78 mg GAE/g. The free radical scavenging activity of CEO (based on IC50) was equal to 9.10 ±â€¯0.63 µg mL-1. In addition, CEO showed a remarkably high inhibitory effect (63%) on ß-carotene bleaching via neutralizing hydroperoxides, which are responsible for the oxidation of highly unsaturated ß-carotene. The antimicrobial effect increased as a function of essential oil concentration. However, there were no inhibitory effects on E. coli at 5 mg mL-1. The electron micrographs demonstrated that CEO caused an increase in the permeabilization of the cells and disrupted the membrane integrity.

Encapsulation of eugenol by spray-drying using whey protein isolate or lecithin: Release kinetics, antioxidant and antimicrobial properties.

The encapsulation of eugenol (E) by spray-drying using whey protein (WP) or soy lecithin (LE) and maltodextrin in combination with oleic acid (OA) and chitosan (CH) was analysed in order to obtain antioxidant and antimicrobial powders for food applications. Formulations with only WP or LE showed higher encapsulation efficiencies (EE) (95-98%) and antibacterial effect against E. coli and L. innocua due to their greater E load. Incorporation of OA or CH promoted lower EE, which negatively affected the antimicrobial and antioxidant activities of the powders. Furthermore, the addition of CH implied less thermal protection against the E losses. The eugenol release was not notably affected by pH or polarity of the food simulant, but the release rate significantly decreased when incorporating OA and CH. The E-LE formulations better retained the eugenol than E-WP powders when heated above 200 °C, this being relevant for the powder inclusion in thermally treated products.

Antilisterial activity of dromedary lactoferrin peptic hydrolysates.

The aim of this study was to explore the antibacterial peptides derived from dromedary lactoferrin (LFc). The LFc was purified from colostrum using a batch procedure with a cation exchange chromatography support and was hydrolyzed with pepsin to generate peptic digest. This peptic digest was fractionated by cation exchange chromatography, and the antilisterial activity of LFc, peptic digest, and obtained fractions was investigated using the bioscreen method. The growth of Listeria innocua ATCC 33090 and LRGIA 01 strains was not inhibited by LFc and its hydrolysates. Two fractions of dromedary lactoferrin peptic hydrolysate were active against both strains. A tandem mass spectroscopy analysis revealed that the 2 active fractions comprised at least 227 different peptides. Among these peptides, 9 found in the first fraction had at least 50% similarity with 10 known antimicrobial peptides (following sequence alignments with the antimicrobial peptide database from the University of Nebraska Medical Center, Omaha). Whereas 9 of these peptides presented homology with honeybee, frog, or amphibian peptides, the 10th peptide, F152SASCVPCVDGKEYPNLCQLCAGTGENKCACSSQEPYFGY192 (specifically found in 1 separated fraction), exibited 54% homology with a synthetic antibacterial peptide (AP00481) derived from human lactoferrin named kaliocin-1. Similarly, the second fraction contained 1 peptide similar to lactoferrampin B, an antibacterial peptide derived from bovine milk. This result suggests that peptic hydrolysis of LFc releases more active antimicrobial peptides than their protein source and thus provides an opportunity for their potential use to improve food safety by inhibiting undesirable and spoilage bacteria.

Antioxidant starch films containing sunflower hull extracts.

This study explores the preparation of antioxidant starch food packaging materials by the incorporation of valuable phenolic compounds extracted from sunflower hulls, which are an abundant by-product from food industry. The phenolic compounds were extracted with aqueous methanol and embedded into starch films. Their effect on starch films was investigated in terms of antioxidant activity, optical, thermal, mechanical, barrier properties and changes in starch molecular structure. The starch molecular structure was affected during thermal processing resulting in a decrease in molar mass, smaller amylopectin molecules and shorter amylose branches. Already 1-2% of extracts were sufficient to produce starch films with high antioxidant capacity. Higher amounts (4-6%) of extract showed the highest antioxidant activity, the lowest oxygen permeability and high stiffness and poor extensibility. The phenolic extracts affected predominantly the mechanical properties, whereas other changes could mainly be correlated to the lower glycerol content which was partially substituted by the extract.

Identification of Peptides Implicated in Antibacterial Activity of Snow Crab Hepatopancreas Hydrolysates by a Bioassay-Guided Fractionation Approach Combined with Mass Spectrometry.

Snow crab (Chionoecetes opilio) by-products are a rich source of biomolecules, such as lipids, proteins, and chitin, which have not been extensively investigated. This study aims to identify antibacterial peptides to enhance the value of C. opilio by-products. After hydrolysis of different component parts using Protamex®, and concentration by solid-phase extraction, the resulting fractions were tested for antibacterial activity against Escherichia coli, Listeria innocua, and Vibrio parahaemolyticus. Hepatopancreas was the only tissue to display antibacterial activity detected using this protocol. Four fractions obtained with and without enzymatic hydrolysis of hepatopancreas followed by SPE C18 fractionation and elution with 50 and 80% acetonitrile demonstrated bacteriostatic activity against L. innocua HPB13, from concentrations of 0.30 to 43.05 mg/mL of peptides/proteins. Eleven peptides sharing at least 80% amino acid homology with four antimicrobial peptides were identified by mass spectrometry. Two peptides had homology to crustin-like and yellowfin tuna GAPDH antimicrobial peptides belonging to the marine organisms Penaeus monodon and Thunnus albacares, respectively. Other peptide sequence homologies were also identified: Odorranain-C7 from the frog Odorrana grahami and a predicted antibacterial peptide in the Asian ladybeetle Harmonia axyridis. These active peptides may represent a novel group of bioactive peptides deserving further investigation as food preservatives.

Prevalence of plasmid-borne benzalkonium chloride resistance cassette bcrABC and cadmium resistance cadA genes in nonpathogenic Listeria spp. isolated from food and food-processing environments.

The sixty-seven nonpathogenic Listeria spp. strains isolated from food and food processing environments in Poland were examined for the presence of benzalkonium chloride (BC) resistance cassette (bcrABC) and four different variants of cadmium resistance determinants (cadA1-cadA4). All the strains were phenotypically resistant to cadmium and 22 among them were also resistant to BC. PCR-based analysis revealed that bcrABC cassette was harbored by 95.5% of the strains phenotypically resistant to BC. All of them harbored also either cadA1 or cadA2 genes (none carried cadA3 or cadA4), which corresponded to the presence of plasmids with two restriction patterns. The strains resistant to cadmium but susceptible to BC harbored only the cadA1 gene variant. DNA-DNA hybridization analysis showed that all the identified bcrABC, cadA1 and cadA2 genes were located within plasmids, classified into 11 groups of RFLP profiles. Only one of the plasmids - pLIS1 of Listeria welshimeri (carrying bcrABC and cadA2) - was capable of efficient conjugal transfer from nonpathogenic Listeria isolates to a pathogenic Listeria monocytogenes strain. Analysis of the complete nucleotide sequence of pLIS1 (the first sequenced plasmid of L. welshimeri species) revealed the presence of genes involved in plasmid replication, stabilization and transfer as well as genes conferring resistance phenotypes. Comparative analysis showed that pLIS1 genome is highly similar to a group of plasmids originating from L. monocytogenes strains. A common feature of pLIS1 and its relatives, besides the presence of the resistance genes, is the presence of numerous transposable elements (TEs). The analysis revealed the important role of TEs in both promoting genetic rearrangements within Listeria spp. plasmids and the acquisition of resistance determinants.

Small Synthetic Peptides Bioconjugated to Hybrid Gold Nanoparticles Destroy Potentially Deadly Bacteria at Submicromolar Concentrations.

Synthetic antibacterial peptides are advanced weapons that scientists design and produce to confront current threats of harmful and mortal pathogens, which could affect humans in everyday life. Recently, many small amino acid sequences, greatly efficient in their antibacterial action, have been reported in the literature. To date, only a few synthetic peptides, acting at micromolar or even tenths of micromolar concentrations, are on the market as commercial products, mainly because of their high cost of production. In this context, materials science can provide fundamental help by engineering small synthetic peptides, powered by hybrid gold nanoparticles, which have been found to strongly enhance antimicrobial activity against bacterial infections. Submicromolar concentrations of the 1018K6 peptide, bioconjugated to hybrid polymer-gold nanoparticles, kill almost 100% of pathogen bacteria, such as Listeria and Salmonella genera, paving the way for economically sustainable commercial products based on this synthetic nanocomplex.