Poloxamer-Based Mixed Micelles Loaded with Thymol or Eugenol for Topical Applications.

Poloxamers (P184, P188, and P407) have been investigated as the carrier system for eugenol or thymol. A synergic effect of mixed Poloxamers was proved by enhanced micellar parameters, with a lower critical micelle concentration (about 0.06 mM) and the highest surface adsorption of 9 × 10-7 mol m-2 for P188/P407. Dynamic light scattering revealed a decrease in micellar size after loading with biomolecules. Three mathematical models were applied to study the release kinetics, of which Korsmeyer-Peppas was the best fitted model. Higher relative release was observed for Poloxamer/eugenol samples, up to a value of 0.8. Poloxamer micelles with thymol were highly influential in bacterial reduction. Single P407/eugenol micelles proved to be bacteriostatic for up to 6 h for S. aureus or up to 48 h for E. coli. Mixed micelles were confirmed to have prolonged bacteriostatic activity for up to 72 h against both bacteria. This trend was also proven by the modified Gompertz model. An optimized P188/P407/eugenol micelle was successfully used as a model system for release study with a particle size of less than 30 nm and high encapsulation efficiency surpassing 90%. The developed mixed micelles were proved to have antibiofilm activity, and thus they provide an innovative approach for controlled release with potential in topical applications.

Non-Alcohol Hand Sanitiser Gels with Mandelic Acid and Essential Oils.

Antimicrobial hand gels have become extremely popular in recent years due to the COVID-19 pandemic. Frequent use of hand sanitising gel can lead to dryness and irritation of the skin. This work focuses on the preparation of antimicrobial acrylic acid (Carbomer)-based gels enhanced by non-traditional compounds-mandelic acid and essential oils-as a substitute for irritating ethanol. Physicochemical properties (pH and viscosity), stability and sensory attributes of the prepared gels were investigated. Antimicrobial activity against representative Gram-positive and Gram-negative bacteria and yeasts was determined. The prepared gels with mandelic acid and essential oil (cinnamon, clove, lemon, and thyme) proved to have antimicrobial activity and even better organoleptic properties than commercial ethanol-based antimicrobial gel. Further, results confirmed that the addition of mandelic acid had a desirable effect on gel properties (antimicrobial, consistency, stability). It has been shown that the essential oil/mandelic acid combination can be a dermatologically beneficial hand sanitiser compared to commercial products. Thus, the produced gels can be used as a natural alternative to alcohol-based daily hand hygiene sanitisers.

Genotypic and Phenotypic Detection of Polyhydroxyalkanoate Production in Bacterial Isolates from Food.

Polyhydroxyalkanoates (PHAs) are widely used in medical and potentially in other applications due to their biocompatibility and biodegradability. Understanding PHA biosynthetic pathways may lead to the detection of appropriate conditions (substrates) for producing a particular PHA type by a specific microbial strain. The aim of this study was to establish a method enabling potentially interesting PHA bacterial producers to be found. In the study, all four classes of PHA synthases and other genes involved in PHA formation (fabG, phaA, phaB, phaG, and phaJ) were detected by PCR in 64 bacterial collection strains and food isolates. Acinetobacter, Bacillus, Cupriavidus, Escherichia, Klebsiella, Lelliottia, Lysinibacillus, Mammaliicoccus, Oceanobacillus, Pantoea, Peribacillus, Priestia, Pseudomonas, Rahnella, Staphylococcus, and Stenotrophomonas genera were found among these strains. Fructose, glucose, sunflower oil, and propionic acid were utilized as carbon sources and PHA production was detected by Sudan black staining, Nile blue staining, and FTIR methods. The class I synthase and phaA genes were the most frequently found, indicating the strains' ability to synthesize PHA from carbohydrates. Among the tested bacterial strains, the Pseudomonas genus was identified as able to utilize all tested carbon sources. The Pseudomonas extremorientalis strain was determined as a prospect for biotechnology applications.

Bioactive zein/chitosan systems loaded with essential oils for food-packaging applications.

BACKGROUND: There has recently been increased interest in biodegradable and sustainable packaging within the food industry. Biopolymer materials based on renewable biomass can be used as alternatives to conventional plastic packaging. A corn protein, zein, possesses excellent film-forming properties because of its hydrophobic nature. It can be used for making edible films and for producing nanofibrous layers. Combination with polysaccharides like chitosan offers promising prospects for the production of delivery systems for the controlled release of active substances. The current trend is to minimize the content of chemical additives; thus essential oils are suitable alternatives to synthetic antimicrobials. RESULTS: This study aimed to develop various zein/chitosan-based film-forming solutions, films, and coatings with antimicrobial substances to prepare active food packaging. Thymol and three essential oils (thyme, cinnamon, oregano) were applied as bioactive ingredients against bacteria, yeasts, and fungi. The incorporation of these natural active compounds led to a decrease in particle size in most film-forming solutions and a reduction of zeta potential compared to controls. Release of the bioactive compound into an aqueous environment was proved by antimicrobial test. A zein/chitosan-based coating with thymol was applied on fresh strawberries. Microbiological analysis over 10 days confirmed the efficient control of bacterial and fungal growth. CONCLUSION: Zein/chitosan (7:1) systems are suitable as bioactive compound carriers to make barriers and to prevent moisture loss, ensuring microbial food quality and prolonging the shelf life of fruits. These systems can serve as sustainable active food packaging. © 2022 Society of Chemical Industry.

Antibacterial and Antifouling Efficiency of Essential Oils-Loaded Electrospun Polyvinylidene Difluoride Membranes.

Nanofibers have become a promising material in many industries in recent years, mainly due to their various properties. The only disadvantage of nanofibers as a potential filtration membrane is their short life due to clogging by bacteria in water treatment. The enrichment of nanofibers with active molecules could prevent these negative effects, represented by essential oils components such as Thymol, Eugenol, Linalool, Cinnamaldehyde and Carvacrol. Our study deals with the preparation of electrospun polyvinylidene difluoride (PVDF)-based nanofibers with incorporated essential oils, their characterization, testing their antibacterial properties and the evaluation of biofilm formation on the membrane surface. The study of the nanofibers' morphology points to the nanofibers' diverse fiber diameters ranging from 570 to 900 nm. Besides that, the nanofibers were detected as hydrophobic material with wettability over 130°. The satisfactory results of PVDF membranes were observed in nanofibers enriched with Thymol and Eugenol that showed their antifouling activity against the tested bacteria Escherichia coli ATCC 25922 and Staphylococcus aureus ATCC 25923. Therefore, these PVDF membranes could find potential applications as filtration membranes in healthcare or the environment.

Antibacterial Filtration Membranes Based on PVDF-co-HFP Nanofibers with the Addition of Medium-Chain 1-Monoacylglycerols.

The efficiency of filtration membranes is substantially lowered by bacterial attachments and potential fouling processes, which reduce their durability and lifecycle. The antibacterial and antifouling properties exhibited by the added materials play a substantial role in their application. We tested a material poly(vinylidene fluoride)-co-hexafluoropropylene (PDVF-co-HFP) based on an electrospun copolymer, where an agent was incorporated with a small amount of ester of glycerol consecutively with caprylic, capric, and lauric acids. Each of these three materials differing in the esters (1-monoacylglycerol, 1-MAG) used was prepared with three weighted concentrations of 1-MAG (1, 2, and 3 wt %). The presence of 1-MAG with an amphiphilic structure resulted in the hydrophilic character of the prepared materials that contributed to the filtration performance. The tested materials (membranes) were characterized with rheological, optical (scanning electron microscopy, SEM), Fourier transform infrared (FTIR) spectroscopy, X-ray diffraction (XRD), and other methods to evaluate antibacterial and antifouling activities. The pure water flux was 6 times higher than that of the neat PVDF-co-HFP membrane when the added 1-MAG attained only 1 wt %. It was experimentally shown that the PVDF-co-HFP/1-MAG membrane with high wettability improved antibacterial activity and antifouling ability. This membrane is highly promising for water treatment due to the safety of antibacterial 1-MAG additives.

Zein-Based Films Containing Monolaurin/Eugenol or Essential Oils with Potential for Bioactive Packaging Application.

Zein is renewable plant protein with valuable film-forming properties that can be used as a packaging material. It is known that the addition of natural cross-linkers can enhance a film's tensile properties. In this study, we aimed to prepare antimicrobial zein-based films enriched with monolaurin, eugenol, oregano, and thyme essential oil. Films were prepared using the solvent casting technique from ethanol solution. Their physicochemical properties were investigated using structural, morphological, and thermal techniques. Polar and dispersive components were analyzed using two models to evaluate the effects on the surface free energy values. The antimicrobial activity was proven using a disk diffusion method and the suppression of bacterial growth was confirmed via a growth kinetics study with the Gompertz function. The films' morphological characteristics led to systems with uniform distribution of essential oils or eugenol droplets combined with a flat-plated structure of monolaurin. A unique combination of polyphenolic eugenol and amphiphilic monoglyceride provided highly stretchable films with enhanced barrier properties and efficiency against Gram-positive and Gram-negative bacteria, yeasts, and molds. The prepared zein-based films with tunable surface properties represent an alternative to non-renewable resources with a potential application as active packaging materials.

Reuse of Textile Waste to Production of the Fibrous Antibacterial Membrane with Filtration Potential.

Wasted synthetic fabrics are a type of textile waste source; the reuse of them brings environmental protection and turns waste into a valuable material. In this work, the used nylon (polyamide) stockings were transmuted into a fine fibrous membrane via an electrospinning process. In addition, the safety antibacterial agent, monoacylglycerol (MAG), was incorporated into a recycled fibrous membrane. The results revealed that the neat, recycled polyamide (rPA) fibers with a hydrophobic surface could be converted into hydrophilic fibers by blending various amounts of MAG with rPA solution prior to electrospinning. The filtration efficiency and air/water vapor permeability of the two types of produced membranes, neat rPA, and rPA/MAG, were tested. Their filtration efficiency (E100) was more than 92% and 96%, respectively. The membranes were classified according to Standard EN1822, and therefore, the membranes rPA and rPA/MAG were assigned to the classes E10 and E11, respectively. The air permeability was not affected by the addition of MAG, and water vapor permeability was slightly enhanced. Based on the obtained data, prepared rPA/MAG fibrous membranes can be evaluated as antifouling against both tested bacterial strains and antimicrobial against S. aureus.

Biofilm Formation Reduction by Eugenol and Thymol on Biodegradable Food Packaging Material.

Biofilm is a structured community of microorganisms adhering to surfaces of various polymeric materials used in food packaging. Microbes in the biofilm may affect food quality. However, the presence of biofilm can ensure biodegradation of discarded packaging. This work aims to evaluate a biofilm formation on the selected biodegradable polymer films: poly (lactic acid) (PLA), poly (butylene adipate-co-terephthalate) (PBAT), and poly (butylene succinate) (PBS) by selected bacterial strains; collection strains of Escherichiacoli, Staphylococcusaureus; and Bacillus pumilus, Bacillussubtilis, Bacillustequilensis, and Stenotrophomonasmaltophilia isolated from dairy products. Three different methods for biofilm evaluation were performed: the Christensen method, 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay, and fluorescence microscopy. High biofilm formation was confirmed on the control PBS film, whereas low biofilm formation ability was observed on the PLA polymer sample. Furthermore, the films with incorporated antimicrobial compounds (thymol or eugenol) were also prepared. Antimicrobial activity and also reduction in biofilm formation on enriched polymer films were determined. Therefore, they were all proved to be antimicrobial and effective in reducing biofilm formation. These films can be used to prepare novel active food packaging for the dairy industry to prevent biofilm formation and enhance food quality and safety in the future.

Novel Polyvinyl Butyral/Monoacylglycerol Nanofibrous Membrane with Antifouling Activity.

Monoacylglycerols (MAGs) have proven of great interest to the foodstuffs industry due to the promising antibacterial activity they show for controlling microbial contamination. Prior to this paper, this antibacterial agent had not been incorporated in a nanofibrous membrane. This study details convenient fabrication of nanofibrous membranes based on polyvinyl butyral (PVB) containing various concentrations of monocaprin (MAG 10) by an electrospinning process. Increasing the concentration of MAG 10 caused differences to appear in the shape of the nanofibers, in addition to which the level of wettability was heightened. Besides exhibiting antibacterial properties, the functional membranes demonstrated especially good antifouling activity. The novel and efficient nanofibrous membranes described have the potential to find eventual application in medical or environmental fields.

Sensitivity to Enterocins of Biogenic Amine-Producing Faecal Enterococci from Ostriches and Pheasants.

Enterococci are widespread bacteria forming the third largest genus among lactic acid bacteria. Some possess probiotic properties or they can produce beneficial proteinaceous antimicrobial substances called enterocins. On the other hand, some enterococci produce biogenic amines (BAs), so this study is focused on the sensitivity to enterocins of biogenic amine-producing faecal enterococci from ostriches and pheasants. Altogether, 60 enterococci isolated from faeces of ostriches and pheasants were tested for production of BAs. This target of the identified enterococci involved 46 strains selected from 140 ostriches and 17 from 60 pheasants involving the species Enterococcus hirae, E. faecium, E. faecalis, and E. mundtii. Although BAs histamine, cadaverine, putrescine, and tryptamine were not detected in the enterococci tested, in general high BA production by the tested enterococci was noted. The species E. hirae formed the majority of the enterococcal strains from ostrichs faeces (34 strains). High production of tyramine (TYM) was measured with an average amount of 958.16 ± 28.18 mg/ml. Among the enterococci from pheasants, the highest was production of TYM compared to phenylethylamine, spermidine, and spermine. Enterococci featured high BA production; however, they were sensitive to seven enterocins with inhibition activity ranging from 100 up to 25,600 AU/ml.

Effect of lantibiotic gallidermin against biogenic amine-producing faecal staphylococci from ostriches and pheasants.

In ostriches and pheasants, there is still limited information relating to staphylococci and their properties. Biogenic amines (BAs) are nitrogenous low-molecular-weight substances with biological functions in animals, plants and microorganisms. In this study, we focused on BA production by targeted faecal staphylococci from ostriches and pheasants and their sensitivity to lantibiotic bacteriocin gallidermin. Gallidermin belongs in a group of polycyclic proteinaceous antimicrobial substances. Thirty-six faecal staphylococci (24 strains from 140 ostriches, 12 from 60 pheasants) comprising different species were tested. Staphylococci from ostriches and pheasants did not produce tryptamine-TRYP, putrescine-PUT, cadaverine-CAD or histamine-HIS. Production of tyramine-TYM, phenylethylamine-PEA was high or very high (100-1000 mg/L). Production of spermine-SPM and spermidine-SPD by staphylococci was very low or low although in the case of staphylococci from pheasants medium production of SPM was found. Because of the risk posed by BAs for consumers, the control of BA-producing bacteria is important from the points of view not only of safety assessment of food-producing animals but also of human health safety. The sensitivity to gallidermin in biogenic amine-producing staphylococci from ostriches and pheasants detected here is the most promising indication for further application of gallidermin for veterinary purposes. The novelty of our study lies in testing the ability of faecal staphylococci from ostriches and pheasants to produce BAs and in their treatment with gallidermin which has so far not been tested in this way.

Decarboxylase-positive Enterococcus faecium strains isolated from rabbit meat and their sensitivity to enterocins.

BACKGROUND: The objective of the study was to determine sensitivity of Enterococcus faecium strains from rabbit meat to enterocins. RESULTS: Twenty-five decarboxylase-positive strains (from rabbit meat) allotted to the species E. faecium by genotypization and by MALDI TOF MS spectrometry identification (evaluation score value range 2.104-2.359; in the range for highly probable species identification-score value 2.300-3.000 and secure probable species identification/probable species identification-2.000-2.299) were studied. Seventeen strains were gelatinase positive. Although they did not produce histamine (HIS), spermidine, and spermine, they produce at least one among seven tested biogenic amines (BAs) in small amounts (2-10 mg/L) or up to very high amounts (>1000 mg/L). Putrescine was produced by two strains. These decarboxylase-positive strains were sensitive to enterocins (Ents). All strains were sensitive to Ent 2019 and Ent 55 (inhibitory activity from 200 to 819 200 AU/mL). Twenty-two strains were inhibited by Ent A(P) and Ent 4231; 20 strains were sensitive to Ent M. CONCLUSION: Our results have spread the basic knowledge related to inhibitory spectrum of enterocins showing sensitivity of decarboxylase-positive strains to enterocins. Protective possibilities of enterocins in meat processing were also indicated.

Decarboxylation activity of enterococci isolated from rabbit meat and staphylococci isolated from trout intestines.

The aim of the study was to explore production of seven biogenic amines (phenylethylamine, histamine, cadaverine, tyramine, putrescine, spermine and/or spermidine) by selected staphylococci and enterococci. Thirty three enterococcal strains isolated from rabbit meat (Oryctolagus cuniculus f. domesticus) and 21 staphylococcal strains isolated from intestinal content of trout (Salmo trutta morpha fario) were tested. Production of biogenic amines was evaluated after cultivation of the tested microorganisms in the de Man, Rogosa and Sharpe Broth (enterococci) or in the Brain Heart Infusion Broth (staphylococci). Both the above cultivation media were enriched with selected amino acids (histidine, tyrosine, arginine, ornithine and lysine; 2g/L each) serving as precursors of biogenic amines. After cultivation, levels of the monitored biogenic amines in broths were analysed by a high performance liquid chromatography equipped with a UV/VIS DAD detector. Among 21 staphylococci, 18 strains produced tyramine or cadaverine, 13 strains formed putrescine or phenylethylamine and only one strain generated histamine. Two staphylococcal strains produced cadaverine levels above 1000 mg/L. Among 33 enterococcal strains, 27 formed cadaverine, 18 strains produced tyramine, 10 strains generated phenylethylamine, and 2 strains gave putrescine. Most of the tyramine producing enterococci generated more than 1000 mg/L of this biogenic amine. Production of spermine or spermidine by the studied strains was not proved.