The use of plant extracts and bacteriophages as an alternative therapy approach in combatting bacterial infections: the study of lytic phages and Stevia rebaudiana.

Introduction: In the light of the problem of antibiotic resistance, the use of combined alternative therapies in combatting bacteria-related disorders has gained popularity. Bacteriophages are one element implemented in new combination therapy. Stevia rebaudiana is known to have antimicrobial activity and regarded as potentially having a synergistic effect with bacteriophages. Therefore, possible interactions of lytic bacteriophages (MS2, T4 and Phi6) with acetone and methanol S. rebaudiana extracts (SRa and SRm) in the bacterial environment were examined. Material and Methods: The interactions were tested using a microdilution method, phage-extract co-incubation assay, static interaction (synography) and dynamic growth profile experiments in a bioreactor. Results: The interactions of the tested factors in a static environment differed from those in a dynamic environment. Dynamic conditions altered the effect of the extracts in a concentration-dependent manner. How different the effect of the SRa extract was to that of the SRm extract on bacterial growth in a dynamic environment depended on the species of the phage and bacterial host. The greatest differences were observed for E. coli strains and their phages, whereas Pseudomonas syringae and the Phi6 phage reacted very similarly to both extracts. Differences also emerged for the same extract in different E. coli strains and their phages. Conclusion: Every extract type should be tested on a case-by-case basis and experiment outcomes should not be generalised before gathering data. Moreover, many varied experiments should be performed, especially when examining such multifactorial mixtures. The tested mixtures could potentially be used in multidrug-resistant bacterial infection treatments.

Polyethylene Films Containing Plant Extracts in the Polymer Matrix as Antibacterial and Antiviral Materials.

Low density polyethylene (LDPE) films covered with active coatings containing mixtures of rosemary, raspberry, and pomegranate CO2 extracts were found to be active against selected bacterial strains that may extend the shelf life of food products. The coatings also offer antiviral activity, due to their influence on the activity of Φ6 bacteriophage, selected as a surrogate for SARS-CoV-2 particles. The mixture of these extracts could be incorporated into a polymer matrix to obtain a foil with antibacterial and antiviral properties. The initial goal of this work was to obtain active LDPE films containing a mixture of CO2 extracts of the aforementioned plants, incorporated into an LDPE matrix via an extrusion process. The second aim of this study was to demonstrate the antibacterial properties of the active films against Gram-positive and Gram-negative bacteria, and to determine the antiviral effect of the modified material on Φ6 bacteriophage. In addition, an analysis was made on the influence of the active mixture on the polymer physicochemical features, e.g., mechanical and thermal properties, as well as its color and transparency. The results of this research indicated that the LDPE film containing a mixture of raspberry, rosemary, and pomegranate CO2 extracts incorporated into an LDPE matrix inhibited the growth of Staphylococcus aureus. This film was also found to be active against Bacillus subtilis. This modified film did not inhibit the growth of Escherichia coli and Pseudomonas syringae cells; however, their number decreased significantly. The LDPE active film was also found to be active against Φ6 particles, meaning that the film had antiviral properties. The incorporation of the mixture of CO2 extracts into the polymer matrix affected its mechanical properties. It was observed that parameters describing mechanical properties decreased, although did not affect the transition of LDPE significantly. Additionally, the modified film exhibited barrier properties towards UV radiation. Modified PE/CO2 extracts films could be applied as a functional food packaging material with antibacterial and antiviral properties.

Double-Layer Agar (DLA) Modifications for the First Step of the Phage-Antibiotic Synergy (PAS) Identification.

The research carried out so far for phage-antibiotic synergy (PAS) differs as regards the technique of modifying the double-layer agar (DLA) method to show the PAS effect on Petri plates, which may contribute to non-uniform research results. Therefore, there is a need to unify the method to effectively detect the PAS effect, at its most basic in vitro test. In this study, bacteriophage T45 and 43 antibiotics belonging to different antibiotic classes were used. Seven different DLA method modifications were tested, in terms of antibiotic addition placement and presence or absence of the base agar. The overall number of phage plaques per plate mainly depended on the antibiotic used. Differences in plaque quantity depended on the type of the DLA method modification. The largest total number of plaques was obtained by the addition of an antibiotic to a bottom agar with the presence of a top agar. This indicates that even though an antibiotic could manifest the PAS effect by a standard disk method, it would be worth examining if the effect is equally satisfactory when applying antibiotics directly into the agar, with regards to using the same bacteriophage and bacterial host.

Zinc Oxide Nanoparticles Enhanced Biomass and Zinc Content and Induced Changes in Biological Properties of Red Perilla frutescens.

The above-ground parts of plants, including leaves, constitute an important part of a human diet. Their mineral and biological composition can be modified by proper preparation of the soil substrate, i.e., supplying it with nutrients. The objective of this study was to assess the influence of zinc oxide nanoparticles (ZnO NPs) at 0, 50, 100 and 200 mg L-1 concentrations on red perilla (Perilla frutescens var. crispa f. purpurea) leaf yield and quality. Plants were grown in 2 L pot under a plastic greenhouse condition. The exposure to ZnO NPs increased leaf fresh and dry weight and leaf Zn content as compared with untreated control. Fresh weight boost was the most pronounced at 50 mg L-1 ZnO NPs. The lowest concentration of ZnO NPs also enhanced the content of total polyphenols, antioxidant activity, and antiradical activity. Treatments with 50 or 100 mg L-1 ZnO NPs boosted the level of total anthocyanins and bacteriostatic activity of 25% extracts. Overall, this study demonstrated that ZnO NPs at low rates is useful as a biostimulant and nanofertilizer for red perilla production.

Packaging Covered with Antiviral and Antibacterial Coatings Based on ZnO Nanoparticles Supplemented with Geraniol and Carvacrol.

The purpose of the study was to obtain an external coating based on nanoparticles of ZnO, carvacrol, and geraniol that could be active against viruses such as SARS-Co-V2. Additionally, the synergistic effect of the chosen substances in coatings was analyzed. The goal of the study was to measure the possible antibacterial activity of the coatings obtained. Testing antiviral activity with human pathogen viruses, such as SARS-Co-V2, requires immense safety measures. Bacteriophages such as phi 6 phage represent good surrogates for the study of airborne viruses. The results of the study indicated that the ZC1 and ZG1 coatings containing an increased amount of geraniol or carvacrol and a very small amount of nanoZnO were found to be active against Gram-positive and Gram-negative bacteria. It is also important that a synergistic effect between these active substances was noted. This explains why polyethylene (PE) films covered with the ZC1 or ZG1 coatings (as internal coatings) were found to be the best packaging materials to extend the quality and freshness of food products. The same coatings may be used as the external coatings with antiviral properties. The ZC1 and ZG1 coatings showed moderate activity against the phi 6 phage that has been selected as a surrogate for viruses such as coronaviruses. It can be assumed that coatings ZG1 and ZC1 will also be active against SARS-CoV-2 that is transmitted via respiratory droplets.

Effects of Gellan Oligosaccharide and NaCl Stress on Growth, Photosynthetic Pigments, Mineral Composition, Antioxidant Capacity and Antimicrobial Activity in Red Perilla.

The growing market demand for plant raw materials with improved biological value promotes the extensive search for new elicitors and biostimulants. Gellan gum derivatives may enhance plant growth and development, but have never been used under stress conditions. Perilla (Perilla frutescens, Lamiaceae) is a source of valuable bioproducts for the pharmaceutical, cosmetic, and food industries. However, there is not much information on the use of biostimulators in perilla cultivation. In this work we investigated the effects of oligo-gellan and salt (100 mM NaCl) on the yield and quality of red perilla (P. frutescens var. crispa f. purpurea) leaves. Plants grown under stress showed inhibited growth, smaller biomass, their leaves contained less nitrogen, phosphorus, potassium, total polyphenol and total anthocyanins, and accumulated considerably more sodium than control plants. Treatment with oligo-gellan under non-saline conditions stimulated plant growth and the fresh weight content of the above-ground parts, enhanced the accumulation of nitrogen, potassium, magnesium and total polyphenols, and increased antioxidant activity as assessed by DPPH and ABTS assays. Oligo-gellan applied under saline conditions clearly alleviated the stress effects by limiting the loss of biomass, macronutrients, and total polyphenols. Additionally, plants pretreated with oligo-gellan and then exposed to 100 mM NaCl accumulated less sodium, produced greater amounts of photosynthetic pigments, and had greater antioxidant activity than NaCl-stressed plants. Irrespective of the experimental treatment, 50% extract effectively inhibited growth of Escherichia coli and Staphylococcus aureus. Both microorganisms were the least affected by 25% extract obtained from plants untreated with either NaCl or oligo-gellan. In conclusion, oligo-gellan promoted plant growth and enhanced the quality of red perilla leaves and efficiently alleviated the negative effects of salt stress.

The Effect of UV Aging on Antimicrobial and Mechanical Properties of PLA Films with Incorporated Zinc Oxide Nanoparticles.

The aim of this study was to examine the influence of accelerated UV-aging on the activity against chosen microorganisms and the mechanical properties of poly-lactic acid (PLA) films enhanced with ZnO nanoparticles. The pure PLA films and tri-layered PLAZnO1%/PLA/PLAZnO1% films of 150 µm thickness were extruded. The samples were treated with UV-A and Q-SUN irradiation. After irradiation the antimicrobial activity and mechanical properties of the films were analyzed. The results of the study demonstrated that PLA films did not inhibit the growth of Staphylococcus aureus, Bacillus cereus, Escherichia coli, Bacillus atrophaeus, and Candida albicans cells. PLA films with incorporated zinc oxide nanoparticles decreased the number of analyzed microorganisms. Accelerated UV aging had no negative effect on the activity of the film containing nano-ZnO against Gram-positive bacteria, but it influenced the activity against Gram-negative cells and C. albicans. Q-SUN irradiation decreased the antimicrobial effect of films with incorporated nanoparticles against B. cereus. UV-A and Q-UV irradiation did not influence the mechanical properties of PLA films containing incorporated ZnO nanoparticles.

A Comparison of the Effects of Packaging Containing Nano ZnO or Polylysine on the Microbial Purity and Texture of Cod (Gadus morhua) Fillets.

Portions of fresh Baltic cod fillets were packed into cellulose boxes (control samples), which were covered with Methyl Hydroxypropyl Celluloses (MHPC) coating with 2% polylysine. The cellulose boxes had square PE films and were enclosed in MHPC coating containing ZnO nanoparticles. The cod fillets were stored at 5 °C and examined after 72 h and 144 h storage times. Results obtained in this study showed that the textural parameters of the cod fillets increased, with both Springiness and Cohesiveness found greater after 144 h of storage for all analysed packaging materials. The Gumminess of fillets increased after storage, but the lowest increase was noted in cod samples that were stored in boxes containing PE films with ZnO nanoparticles. It was found that water loss from the cod fillets in these boxes was also lowest. The Adhesiveness of the fish samples stored in boxes devoid of active coatings also increased. In contrast to the packaging material devoid of active coatings, the storage of fillets in active coating boxes resulted in a decrease of adhesiveness. Microbial analysis showed that packaging material containing nano-ZnO was found to be more active against mesophilic and psychotropic bacterial cells than the coatings with polylysine after 72 h and 144 h of storage.

Exopolysaccharide Gellan Gum and Derived Oligo-Gellan Enhance Growth and Antimicrobial Activity in Eucomis Plants.

One of the visible trends in the cultivation of plants, particularly of medicinal ones, is the increasing interest of researchers in polysaccharides and their derivatives that show biostimulatory properties and are also safe to use. In the current study, we evaluated the effects of gellan gum and its depolymerized form oligo-gellan, on growth and antimicrobial activity of two ornamental species Eucomis bicolor and Eucomis comosa used in natural medicine. The biopolymers were applied in the form of bulb coating prepared by using polyelectrolyte complexes. In both species investigated, gellan gum and oligo-gellan enhanced the fresh weight of leaves and bulbs, the performance of the photosynthetic apparatus, and the leaf content of basic macronutrients. In comparison with the control, the plants treated with oligo-gellan accumulated more biomass, were first to flower, and had the highest leaf content of potassium. The extracts from the bulbs treated with gellan gum and oligo-gellan showed higher effectiveness in reducing the count of Bacillus atrophaeus, Escherichia coli, and Staphylococcus aureus than those from the bulbs not treated with the polysaccharides. The research described here largely expands our current knowledge on the effects of gellan gum derivatives and has a huge practical potential in agriculture production.

New Poly(lactic acid) Active Packaging Composite Films Incorporated with Fungal Melanin.

In this work, fungal melanin was used for the first time to prepare poly(lactic acid)-based composites. The films of various melanin concentrations (0.025%, 0.05% and 0.2% w/w) were prepared using an extrusion method. The mechanical, antioxidant, antimicrobial, water vapor and UV-Vis barrier properties, as well as available polyphenolics on the surface, were studied. FT-IR and Raman spectroscopy studies were carried out to analyze the chemical composition of the resulting films. The hydrophobicity, color response, thermal, optical properties, and opacity values were also determined. The results of this study show that the addition of fungal melanin to poly(lactic acid) (PLA) as a modifier influenced mechanical and water vapor barrier properties depending on melanin concentration. In low concentration, melanin enhanced the mechanical and barrier properties of the modified films, but in larger amounts, the properties were decreased. The UV-Vis barrier properties of PLA/melanin composites were marginally improved. Differential Scanning Calorimetry (DSC) analysis indicated that crystallinity of PLA increased by the addition of melanin, but this did not affect the thermal stability of the films. Modified PLA/melanin films showed good antioxidant activity and were active against Enterococcus faecalis, Pseudomonas aeruginosa and Pseudomonas putida. The addition of melanin caused changes in color values, decreasing lightness and increasing the redness and yellowness of films. Based on the results of this study, fungal melanin has good potential to be exploited as a value-added modifier that can improve the overall properties of PLA.

The Influence of Accelerated UV-A and Q-SUN Irradiation on the Antibacterial Properties of Hydrophobic Coatings Containing Eucomis comosa Extract.

The purpose of this research was to examine the antimicrobial properties against Gram-positive bacteria, as well as the water vapour characteristic of polylactic acid (PLA) films covered with a methyl⁻hydroxypropyl⁻cellulose (MHPC)/cocoa butter carrier containing Eucomis comosa extract as an active substance. The second purpose of the study was to evaluate the influence of accelerated UV-A and Q-SUN irradiation (UV-aging) on the antimicrobial properties and the barrier characteristic of the coatings. The results of the study revealed that MHPC/cocoa butter coatings had no influence on the growth of Staphylococcus aureus, Bacillus cereus, and Bacillus atrophaeus. MHPC/cocoa butter coatings containing E. comosa extract reduced the number of bacterial strains. MHPC/cocoa butter coatings also decreased the water vapour permeability of PLA. It was shown that accelerated UV-A and Q-SUN irradiations altered the chemical composition of the coatings containing cocoa butter. Despite the alteration of the chemical composition of the layers, the accelerated Q-SUN and UV-A irradiation had no influence on the antimicrobial properties of E. comosa extract coatings against S. aureus and B. cereus. It was found that only Q-SUN irradiation decreased the coating activity with an extract against B. atrophaeus, though this was to a small degree.

Antimicrobial activity of water and acetone extracts of some Eucomis taxa.

OBJECTIVE: To evaluate the antibacterial and antifungal properties of acetone and water extracts of selected Eucomis taxa. METHODS: The bulbs of Eucomis bicolor, Eucomis comosa (E. comosa) and E. comosa cv. were dried and examples from each experimental treatment were ground separately into powder. Each sample was divided into two groups with equal amounts of powder, and respectively extracted with water and a 70% solution of aqueous acetone. The crude water extracts were then filtered through a 0.2 µm filter. The 70% aqueous acetone extracts were next concentrated at 40 °C. After the evaporation of the acetone, the samples were additionally filtered through a 0.2 µm filter. The antibacterial and antifungal activities of the extracts against chosen microorganisms were then determined. RESULTS: The results of the study demonstrated that the water and acetone extracts of Eucomis bulbs have an influence on the viability of Staphylococcus aureus and Bacillus atrophaeus strains. In the case of mediums containing E. comosa and E. comosa cv. extracts, a decrease in the number of gram-positive bacteria was dependent on the extract concentration. The best results were obtained in the case of the 25% extracts. The water and acetone extracts of Eucomis bulbs did not cause a decrease in the number of Escherichia coli cells. Additionally, antifungal activity against Aspergillus niger, Botrytis cinerea, Stachybotrys chartarum, Mucor circinelloides and Rhizopus oryzae cells were also not observed. CONCLUSIONS: The tested gram-negative and fungi microorganisms show resistance towards acetone and water extracts of Eucomis bulbs. The highest activity is found in the case of Aspergillus clavatus, Staphylococcus aureus and Bacillus atrophaeus strains, which shows water and acetone extracts of 25%.

The Influence of Accelerated UV-A and Q-SUN Irradiation on the Antimicrobial Properties of Coatings Containing ZnO Nanoparticles.

The influence of accelerated UV-A and Q-SUN irradiation on the antimicrobial properties of coatings containing ZnO nanoparticles was investigated using a polyethylene (PE) film covering. The results of the study showed that Methyl Hydroxypropyl Celluloses (MHPC) coatings did not influence the growth of S. aureus, B. cereus, E. coli, P. aeruginosa or C. albicans cells. MHPC coatings containing ZnO nanoparticles inhibited the growth of bacterial strains and reduced the number of C. albicans strains. Accelerated Q-SUN and UV-A irradiation had no influence on the antimicrobial effect of nano ZnO coatings against S. aureus, B. cereus and E. coli. Q-SUN irradiation decreased the activity of MHPC coatings containing nanoparticles against P. aeruginosa and C. albicans. An FT-IR analysis clearly showed that ZnO nanoparticles shielded the MHPC coating during Q-SUN irradiation.