Biological activity of silver nanoparticles synthesized using viticultural waste.

This research paper presents a novel approach to the green synthesis of silver nanoparticles (AgNPs) using viticultural waste, allowing to obtain NP dispersions with distinct properties and morphologies (monodisperse and polydisperse AgNPs, referred to as mAgNPs and pAgNPs) and to compare their biological activities. Our synthesis method utilized the ethanolic extract of Vitis vinifera pruning residues, resulting in the production of mAgNPs and pAgNPs with average sizes of 12 ± 5 nm and 19 ± 14 nm, respectively. Both these AgNPs preparations demonstrated an exceptional stability in terms of size distribution, which was maintained for one year. Antimicrobial testing revealed that both types of AgNPs inhibited either the growth of planktonic cells or the metabolic activity of biofilm sessile cells in Gram-negative bacteria and yeasts. No comparable activity was found towards Gram-positives. Overall, pAgNPs exhibited a higher antimicrobial efficacy compared to their monodisperse counterparts, suggesting that their size and shape may provide a broader spectrum of interactions with target cells. Both AgNP preparations showed no cytotoxicity towards a human keratinocyte cell line. Furthermore, in vivo tests using a silkworm animal model indicated the biocompatibility of the phytosynthesized AgNPs, as they had no adverse effects on insect larvae viability. These findings emphasize the potential of targeted AgNPs synthesized from viticultural waste as environmentally friendly antimicrobial agents with minimal impact on higher organisms.

Synthesis and Characterization of Lignin-Silver Nanoparticles.

Metal nanoparticle synthesis via environmentally friendly methods is gaining interest for their potential advantages over conventional physico-chemical approaches. Herein, we propose a robust green synthesis route for lignin-modified silver nanoparticles, utilizing the recovery of lignin as a renewable raw material and exploring its application in valuable areas. Through a systematic approach combining UV-Vis spectroscopy with AAS and DLS, we identified repeatable and scalable reaction conditions in an aqueous solution at pH 11 for homogeneous silver nanoparticles with high uniformity. The TEM median sizes ranged from 12 to 15 nm with circularity between 0.985 and 0.993. The silver nanoparticles yield exceeded 0.010 mol L-1, comparable with traditional physico-chemical methods, with a minimal loss of silver precursor ranging between 0.5 and 3.9%. Characterization by XRD and XPS revealed the presence of Ag-O bonding involving lignin functional groups on the pure face-centered cubic structure of metallic silver. Moreover, the lignin-modified silver nanoparticles generated a localized thermal effect upon near-infrared laser irradiation (808 nm), potentially allowing for targeted applications in the biomedical field. Our study showcases the potential of lignin as a renewable reducing and capping agent for silver nanoparticle synthesis, addressing some shortcomings of green synthesis approaches and contributing to the development of suitable nanomaterials.

Use of non-thermal plasma pre-treatment to enhance antibiotic action against mature Pseudomonas aeruginosa biofilms.

Non-thermal plasma (NTP), generated at atmospheric pressure by DC cometary discharge with a metallic grid, and antibiotics (gentamicin-GTM, ceftazidime-CFZ and polymyxin B-PMB), either alone or in combination, were used to eradicate the mature biofilm of Pseudomonas aeruginosa formed on Ti-6Al-4V alloy. Our aim was to find the conditions for NTP pre-treatment capable of enhancing the action of the antibiotics and thus reducing their effective concentrations. The NTP treatment increased the efficacy of relatively low concentrations of antibiotics. Generally, the highest effect was achieved with GTM, which was able to suppress the metabolic activity of pre-formed P. aeruginosa biofilms in the concentration range of 4-9 mg/L by up to 99%. In addition, an apparent decrease of biofilm-covered area was confirmed after combined NTP treatment and GTM action by SYTO®13 staining using fluorescence microscopy. Scanning electron microscopy confirmed a complete eradication of P. aeruginosa ATCC 15442 mature biofilm from Ti-6Al-4V alloy when using 0.25 h NTP treatment and subsequent treatment by 8.5 mg/L GTM. Therefore, NTP may be used as a suitable antibiofilm agent in combination with antibiotics for the treatment of biofilm-associated infections caused by this pathogen.

Aspergillus fumigatus DBM 4057 biofilm formation is inhibited by chitosan, in contrast to baicalein and rhamnolipid.

The biofilms of filamentous-forming fungi are a novel and still insufficiently understood research topic. We have studied Aspergillus fumigatus, an ubiquitous opportunistic pathogenic fungus, as a representative model for a study of biofilm formation by filamentous fungi and for assessing the potential anti-biofilm activity of natural substances. The activity of antibiotic amphotericin B and selected natural substances: baicalein, chitosan and rhamnolipid was studied. The minimum suspension inhibitory concentrations (MIC) were determined and the biofilm susceptibility was investigated by determining the metabolic activity of sessile cells (XTT assay) and total biofilm biomass (crystal violet staining). Significant time-dependent differences in substances' anti-biofilm activity were observed. Images of A. fumigatus biofilm were obtained by Cellavista automatic light microscope and spinning disc confocal microscopy. Baicalein and rhamnolipid were not found as suitable substances for inhibition of the A. fumigatus biofilm formation, as neither of the substances inhibited the sessile cells metabolic activity or the total biofilm biomass even at tenfold MIC after 48 h. In contrast, chitosan at 10 × MIC (25 µg mL-1), suppressed the biofilm metabolic activity by 90 % and the total biofilm biomass by 80 % even after 72 h of cultivation. Amphotericin B inhibited only 14 % of total biofilm biomass (crystal violet staining) and 35 % of metabolic activity (XTT assay) of adherent cells under the same conditions. Our results therefore suggest chitosan as potential alternative for treating A. fumigatus biofilm-associated infections.

Influencing fatty acid composition of yeasts by lanthanides.

The growth of microorganisms is affected by cultivation conditions, concentration of carbon and nitrogen sources and the presence of trace elements. One of the new possibilities of influencing the production of cell mass or lipids is the use of lanthanides. Lanthanides are biologically non-essential elements with wide applications in technology and industry and their concentration as environmental contaminants is therefore increasing. Although non-essential, lanthanides have been proposed (and even used) to produce beneficial effects in plants but their mechanisms of action are unclear. Recently, it was suggested that they may replace essential elements or operate as potent blockers of Ca(2+) channels. We tested the effect of low concentrations of lanthanides on traditional biotechnologically useful yeast species (Kluyveromyces polysporus, Saccharomyces cerevisiae, Torulospora delbrueckii), and species capable of high accumulation of lipids (Rhodotorula glutinis, Trichosporon cutaneum, Candida sp., Yarrowia lipolytica). Low concentrations of lanthanum and monazite were conducive to an increase in cell mass and lipids and also higher production of palmitoleic acid, commonly used in cosmetics and medicine, and ω6-linoleic acid which is a precursor of thromboxanes, prostaglandins and leucotrienes.

Exploring the antimicrobial potential of chitosan nanoparticles: synthesis, characterization and impact on Pseudomonas aeruginosa virulence factors.

The escalating antibiotic resistance observed in bacteria poses a significant threat to society, with the global prevalence of resistant strains of Pseudomonas aeruginosa on the rise. Addressing this challenge necessitates exploring strategies that would complement existing antimicrobial agents, e.g. by substances mitigating bacterial virulence without eliciting selective pressure for resistance emergence. In this respect, free-form chitosan has demonstrated promising efficacy, prompting our investigation into reinforcing its effects through nanoparticle formulations. Our study focuses on the preparation of chitosan nanoparticles under suitable conditions while emphasizing the challenges associated with stability that can affect biological activity. These challenges are mitigated by introducing quaternized chitosan, which ensures colloidal stability in the culture media. Our approach led to the production of trimethylchitosan nanoparticles with a median size of 103 nm, circularity of 0.967, and a charge of 14.9 ± 3.1 mV, stable within a one-month period in a water stock solution, showing promising attributes for further valorization. Furthermore, the study delves into the antimicrobial activity of trimethylchitosan nanoparticles on Pseudomonas aeruginosa and confirms the benefits of both nanoformulation and modification of chitosan, as our prepared nanoparticles inhibit 50% of the bacterial population at concentration ≥160 mg L-1 within tested strains. Additionally, we identified a concentration of 5 mg L-1 that no longer impedes bacterial growth, allowing reliable verification of the effect of the prepared nanoparticles on Pseudomonas aeruginosa virulence factors, including motility, protease activity, hemolytic activity, rhamnolipids, pyocyanin, and biofilm production. Although trimethylchitosan nanoparticles exhibit promise as an effective antibiofilm agent (reducing biofilm development by 50% at concentrations ranging from 80 to 160 mg L-1) their impact on virulence manifestation is likely not directly associated with quorum sensing. Instead, it can probably be attributed to non-specific interactions with the bacterial surface. This exploration provides valuable insights into the potential of quaternized chitosan nanoparticles in addressing Pseudomonas aeruginosa infections and underscores the multifaceted nature of their antimicrobial effects.

Isolation and characterization of multiple-stress tolerant bacteria from radon springs.

Radon springs, characterized by their high concentrations of radon gas (Rn222), are extreme environments with unique physicochemical conditions distinct from conventional aquatic ecosystems. Our research aimed to investigate microbial life in radon springs, focusing on isolating extremophilic bacteria and assessing their resistance to adverse conditions. Our study revealed the prevalence of Actinomycetia species in the radon spring environment. We conducted various tests to evaluate the resistance of these isolates to oxidative stress, irradiation, desiccation, and metal ion content. These extremophilic bacteria showed overall higher resistance to these stresses compared to control strains. Lipidomic analysis was also employed to provide insights into the adaptive mechanisms of these bacteria which were found mainly in the correlations among individual clusters and changes in content of fatty acids (FA) as well as differences between content and type of FAs of environmental isolates and type strains.

Comparative assessment of UV-C radiation and non-thermal plasma for inactivation of foodborne fungal spores suspension in vitro.

Fungal contamination poses a persistent challenge to industries, particularly in food, healthcare, and clinical sectors, due to the remarkable resilience of fungi in withstanding conventional control methods. In this context, our research delves into the comparative efficacy of UV radiation and non-thermal plasma (NTP) on key foodborne fungal contaminants - Alternaria alternata, Aspergillus niger, Fusarium culmorum, and Fusarium graminearum. The study examined the impact of varying doses of UV radiation on the asexual spores of all mentioned fungal strains. Simultaneously, the study compared the effects of UV radiation and NTP on the metabolic activity of cells after spore germination and their subsequent germination ability. The results revealed that UV-C radiation (254 nm) did not significantly suppress the metabolic activity of cells after spore germination. In contrast, NTP exhibited almost 100% effectiveness on both selected spores and their subsequent germination, except for A. niger. In the case of A. niger, the effectiveness of UV-C and NTP was nearly comparable, showing only a 35% decrease in metabolic activity after 48 hours of germination, while the other strains (A. alternata, F. culmorum, F. graminearum) exhibited a reduction of more than 95%. SEM images illustrate the morphological changes in structure of all tested spores after both treatments. This study addresses a crucial gap in existing literature, offering insights into the adaptation possibilities of treated cells and emphasizing the importance of considering exposure duration and nutrient conditions (introduction of fresh medium). The results highlighted the promising antimicrobial potential of NTP, especially for filamentous fungi, paving the way for enhanced sanitation processes with diverse applications.

Detection of microscopic filamentous fungal biofilms - Choosing the suitable methodology.

Microscopic filamentous fungi are ubiquitous microorganisms that adapt very easily to a variety of environmental conditions. Due to this adaptability, they can colonize a number of various surfaces where they are able to start forming biofilms. Life in the form of biofilms provides them with many benefits (increased resistance to desiccation, UV radiation, antimicrobial compounds, and host immune response). The aim of this study is to find a reliable and reproducible methodology to determine biofilm growth of selected microscopic filamentous fungi strains. Several methods (crystal violet staining, MTT assay, XTT assay, resazurin assay) for the determination of total biofilm biomass and its metabolic activity were tested on four fungi - Alternaria alternata, Aspergillus niger, Fusarium culmorum and Fusarium graminearum, and their biofilm was also imaged by spinning disc confocal microscopy using fluorescent dyes. A reproducible biofilm quantification method is essential for the subsequent testing of the biofilm growth suppression using antifungal agents or physical methods. Crystal violet staining was found to be a suitable method for the determination of total biofilm biomass of selected strains, and the MTT assay for the determination of metabolic activity of the biofilms. Calcofluor white and Nile red fluorescent stains successfully dyed the hyphae of microscopic fungi.

Silver Nanoparticle Production Mediated by Vitis vinifera Cane Extract: Characterization and Antibacterial Activity Evaluation.

The ever-growing range of possible applications of nanoparticles requires their mass production. However, there are problems resulting from the prevalent methods of nanoparticle production; physico-chemical routes of nanoparticle synthesis are not very environmentally friendly nor cost-effective. Due to this, the scientific community started exploring new methods of nanoparticle assembly with the aid of biological agents. In this study, ethanolic Vitis vinifera cane extract combined with silver nitrate was used to produce silver nanoparticles. These were subsequently characterized using UV-visible (UV-Vis) spectrometry, transmission electron microscopy, and dynamic light-scattering analysis. The antimicrobial activity of produced nanoparticles was tested against the planktonic cells of five strains of Gram-negative bacterium Pseudomonas aeruginosa (PAO1, ATCC 10145, ATCC 15442, DBM 3081, and DBM 3777). After that, bactericidal activity was assessed using solid medium cultivation. In the end, nanoparticles' inhibitory effect on adhering cells was analyzed by measuring changes in metabolic activity (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay-MTT). Our results confirmed that ethanolic Vitis vinifera cane extract is capable of mediating silver nanoparticle production; synthesis was conducted using 10% of extract and 1 mM of silver nitrate. The silver nanoparticles' Z-average was 68.2 d nm, and their zeta potential was -30.4 mV. These silver nanoparticles effectively inhibited planktonic cells of all P. aeruginosa strains in concentrations less than 5% v/v and inhibited biofilm formation in concentrations less than 6% v/v. Moreover, minimum bactericidal concentration was observed to be in the range of 10-16% v/v. According to the results in this study, the use of wine agriculture waste is an ecological and economical method for the production of silver nanoparticles exhibiting significant antimicrobial properties.

Cobalt Bis-Dicarbollide Enhances Antibiotics Action towards Staphylococcus epidermidis Planktonic Growth Due to Cell Envelopes Disruption.

The emergence of antibiotic resistance in opportunistic pathogens represents a huge problem, the solution for which may be a treatment with a combination of multiple antimicrobial agents. Sodium salt of cobalt bis-dicarbollide (COSAN.Na) is one of the very stable, low-toxic, amphiphilic boron-rich sandwich complex heteroboranes. This compound has a wide range of potential applications in the biological sciences due to its antitumor, anti-HIV-1, antimicrobial and antibiofilm activity. Our study confirmed the ability of COSAN.Na (in the concentration range 0.2-2.48 µg/mL) to enhance tetracycline, erythromycin, and vancomycin action towards Staphylococcus epidermidis planktonic growth with an additive or synergistic effect (e.g., the combination of 1.24 µg/mL COSAN.Na and 6.5 µg/mL TET). The effective inhibitory concentration of antibiotics was reduced up to tenfold most efficiently in the case of tetracycline (from 65 to 6.5 µg/mL). In addition, strong effect of COSAN.Na on disruption of the cell envelopes was determined using propidium iodide uptake measurement and further confirmed by transmission electron microscopy. The combination of amphiphilic COSAN.Na with antibiotics can therefore be considered a promising way to overcome antibiotic resistance in Gram-positive cocci.

Production and Characterization of Rhamnolipids Produced by Pseudomonas aeruginosa DBM 3774: Response Surface Methodology Approach.

Rhamnolipids are extensively studied biosurfactants due to their potential in many industrial applications, eco-friendly production and properties. However, their availability for broader application is severely limited by their production costs, therefore the optimization of efficacy of their cultivation gains significance as well as the information regarding the physio-chemical properties of rhamnolipids resulting from various cultivation strategies. In this work, the bioprocess design focused on optimization of the rhamnolipid yield of Pseudomonas aeruginosa DBM 3774 utilizing the response surface methodology (RSM). Six carbon sources were investigated for their effect on the rhamnolipid production. The RSM prediction improved the total rhamnolipid yield from 2.2 to 13.5 g/L and the rhamnolipid productivity from 11.6 to 45.3 mg/L/h. A significant effect of the carbon source type, concentration and the C/N ratio on the composition of the rhamnolipid congeners has been demonstrated for cultivation of P. aeruginosa DBM 3774 in batch cultivation. Especially, changes in presence of saturated fatty acid in the rhamnolipid congeners, ranging from 18.8% of unsaturated fatty acids (carbon source glycerol; 40 g/L) to 0% (sodium citrate 20 g/L) were observed. This demonstrates possibilities of model based systems as basis in cultivation of industrially important compounds like biosurfactants rhamnolipids and the importance of detailed study of interconnection between cultivation conditions and rhamnolipid mixture composition and properties.

Antimicrobial properties and applications of metal nanoparticles biosynthesized by green methods.

There is a growing interest in the potential and application of metal nanoparticles across many fields. A vast array of techniques for metal nanoparticle synthesis has been discovered; however, sustainability, cost-effectiveness, and environmental concerns favor the green biological approach, using various plant and microbial sources. This review describes the diversity in green methods for nanoparticle biosynthesis, antimicrobial properties of metal nanoparticles and their potential applications. Metal nanoparticle biosynthesis by extracts and solutions obtained from plants, bacteria, fungi and templates such as viruses are discussed. As biosynthesized nanoparticles have been proven to possess antibacterial, antifungal, and even antiviral properties, these are discussed in detail, with silver and gold nanoparticles as the most studied and with the highest potential for medical application. The focus on prospective antimicrobial applications of nanoparticles stems from the arising resistance of many serious pathogens to traditional disinfectants and antibiotics. Other fields for the application of biosynthesized nanoparticles are also stated briefly, such as in agriculture as pesticides, in wastewater treatment and bioremediation. Finally, the limitations and safety issues connected with widespread use of nanoparticles are discussed.

Antibiofilm activity of silver nanoparticles biosynthesized using viticultural waste.

Green methods have become vital for sustainable development of the scientific and commercial sphere; however, they can bring new challenges, including the need for detailed characterization and elucidation of efficacy of their products. In this study, green method of silver nanoparticles (AgNPs) production was employed using an extract from grapevine canes. The aim of the study was to contribute to the knowledge about biosynthesized AgNPs by focusing on elucidation of their antifungal efficiency based on their size and/or hypothesized synergy with bioactive substances from Vitis vinifera cane extract. The antifungal activity of AgNPs capped and stabilized with bioactive compounds was tested against the opportunistic pathogenic yeast Candida albicans. Two dispersions of nanoparticles with different morphology (characterized by SEM-in-STEM, DLS, UV-Vis, XRD, and AAS) were prepared by modification of reaction conditions suitable for economical production and their long-term stability monitored for six months was confirmed. The aims of the study included the comparison of the antifungal effect against suspension cells and biofilm of small monodisperse AgNPs with narrow size distribution and large polydisperse AgNPs. The hypothesis of synergistic interaction of biologically active molecules from V. vinifera extracts and AgNPs against both cell forms were tested. The interactions of all AgNPs dispersions with the cell surface and changes in cell morphology were imaged using SEM. All variants of AgNPs dispersions were found to be active against suspension and biofilm cells of C. albicans; nevertheless, surprisingly, larger polydisperse AgNPs were found to be more effective. Synergistic action of nanoparticles with biologically active extract compounds was proven for biofilm cells (MBIC80 20 mg/L of polydisperse AgNPs in extract), while isolated nanoparticles suspended in water were more active against suspension cells (MIC 20 mg/L of polydisperse AgNPs dispersed in water). Our results bring new insight into the economical production of AgNPs with defined characteristics, which were proven to target a specific mode of growth of significant pathogen C. albicans.

Analysis of Bacteriohopanoids from Thermophilic Bacteria by Liquid Chromatography-Mass Spectrometry.

Background: Hopanoids modify plasma membrane properties in bacteria and are often compared to sterols that modulate membrane fluidity in eukaryotes. In some microorganisms, they can also allow adaptations to extreme environments. Methods: Hopanoids were identified by liquid chromatography-mass spectrometry in fourteen strains of thermophilic bacteria belonging to five genera, i.e., Alicyclobacillus, Brevibacillus, Geobacillus, Meiothermus, and Thermus. The bacteria were cultivated at temperatures from 42 to 70 °C. Results: Regardless of the source of origin, the strains have the same tendency to adapt the hopanoid content depending on the cultivation temperature. In the case of aminopentol, its content increases; aminotetrol does not show a significant change; and in the case of aminotriol the content decreases by almost a third. The content of bacteriohopanetetrol and bacteriohopanetetrol glycoside decreases with increasing temperature, while in the case of adenosylhopane the opposite trend was found. Conclusions: Changes in hopanoid content can be explained by increased biosynthesis, where adenosylhopane is the first intermediate in the biosynthesis of the hopanoid side chain.

Low-molecular weight chitosan enhances antibacterial effect of antibiotics and permeabilizes cytoplasmic membrane of Staphylococcus epidermidis biofilm cells.

This study evaluated the effect of low-molecular weight chitosan on Staphylococcus epidermidis, a common colonizer of joint implants and other prosthetic devices. We have also attempted to elucidate its mechanism of action. Chitosan was found to be effective against both the planktonic and biofilm cells (MIC80 35-40 mg/L; MBIC80 40-150 mg/L), in contrast to the antibiotics erythromycin and tetracycline with no antibiofilm activity (MBIC80 not found). In combination, chitosan had an additive effect with antibiotics on suspension growth of S. epidermidis (FICi 0.7-1.0), and the combinatory action caused a complete inhibition of biofilm metabolic activity in some cases. In addition, chitosan caused rapid cellular damage and enhanced antihaemolytic activity of tetracycline in combination towards S. epidermidis biofilm cells. Chitosan efficiently inhibited S. epidermidis growth acting via cell membrane damage, yet the extent of antimicrobial and antibiofilm activities was quite strain-specific. It was proved to be a very efficient antimicrobial agent worth further examination as a potent candidate in pharmaceutical research. Apart from antimicrobial activity, it also acted as antivirulence enhancing agent which is a very promising strategy for alternative infectious diseases treatment.

Composition and Biological Activity of Vitis vinifera Winter Cane Extract on Candida Biofilm.

Vitis vinifera canes are waste material of grapevine pruning and thus represent cheap source of high-value polyphenols. In view of the fact that resistance of many pathogenic microorganisms to antibiotics is a growing problem, the antimicrobial activity of plant polyphenols is studied as one of the possible approaches. We have investigated the total phenolic content, composition, antioxidant activity, and antifungal activity against Candida biofilm of an extract from winter canes and a commercially available extract from blue grapes. Light microscopy and confocal microscopy imaging as well as crystal violet staining were used to quantify and visualize the biofilm. We found a decrease in cell adhesion to the surface depending on the concentration of resveratrol in the cane extract. The biofilm formation was observed as metabolic activity of Candida albicans, Candida parapsilosis and Candida krusei biofilm cells and the minimum biofilm inhibitory concentrations were determined. The highest inhibition of metabolic activity was observed in Candida albicans biofilm after treatment with the cane extract (30 mg/L) and blue grape extract (50 mg/L). The composition of cane extract was analyzed and found to be comparatively different from blue grape extract. In addition, the content of total phenolic groups in cane extract was three-times higher (12.75 gGA/L). The results showed that cane extract was more effective in preventing biofilm formation than blue grape extract and winter canes have proven to be a potential source of polyphenols for antimicrobial and antibiofilm treatment.

Rhamnolipids as a Tool for Eradication of Trichosporon cutaneum Biofilm.

Microbial biofilms formed by pathogenic and antibiotic-resistant microorganisms represent a serious threat for public health in medicine and many industrial branches. Biofilms are involved in many persistent and chronic infections, the biofouling of water and food contamination. Therefore, current research is involved in the development of new treatment strategies. Biofilm is a complex system, and thus all aspects of the measurement and monitoring of its growth and eradication in various conditions, including static and dynamic flow, are issues of great importance. The antibiofilm character of rhamnolipid mixtures produced by four Pseudomonas aeruginosa strains was studied under different conditions. For this purpose, the biofilm of opportunistic pathogen Trichosporon cutaneum was used and treated under static conditions (microscope glass coverslip in a Petri dish) and under dynamic conditions (a single-channel flow cell). The results show that the biological activity of rhamnolipids depends both on their properties and on the conditions of the biofilm formation. Therefore, this aspect must be taken into account when planning the experimental or application design.

A novel approach for the production of green biosurfactant from Pseudomonas aeruginosa using renewable forest biomass.

The rising demand for surfactants by the pharmaceuticals and cosmetic industries has generated vast amounts of petroleum-based synthetic surfactants, which are often toxic and non-degradable. Owing to their low toxicity, stability in extreme conditions, and biodegradability, biosurfactants could represent a sustainable alternative. The present study aimed to maximize the production of rhamnolipids (RL) from Pseudomonas aeruginosa by optimizing glucose concentration, temperature, and C/N and C/P ratios. After 96 h of cultivation at 37 °C, the final RL concentration was 4.18 ± 0.19 g/L with a final yield of 0.214 ± 0.010 g/gglucose when pure glucose was used as a carbon source. At present, the main obstacle towards commercialization of RL production is economic sustainability, due to the high cost of downstream processes and media components. For this reason, a renewable source such as wood hydrolysates (from birch and spruce woodchips) was examined here as a possible source of glucose for RL production. Both hydrolysates proved to be adequate, resulting in 2.34 ± 0.17 and 2.31 ± 0.10 g/L of RL, respectively, and corresponding yields of 0.081 ± 0.006 and 0.089 ± 0.004 g/gsugar after 96 h. These results demonstrate the potential of using renewable biomass for the production of biosurfactants and, to the best of our knowledge, they constitute the first report on the use of wood hydrolysates for RL production.

Effect of resveratrol and Regrapex-R-forte on Trichosporon cutaneum biofilm.

Microorganisms that cause chronic infections exist predominantly as surface-attached stable communities known as biofilms. Microbial cells in biofilms are highly resistant to conventional antibiotics and other forms of antimicrobial treatment; therefore, modern medicine tries to develop new drugs that exhibit anti-biofilm activity. We investigated the influence of a plant polyphenolic compound resveratrol (representative of the stilbene family) on the opportunistic pathogen Trichosporon cutaneum. Besides the influence on the planktonic cells of T. cutaneum, the ability to inhibit biofilm formation and to eradicate mature biofilm was studied. We have tested resveratrol as pure compound, as well as resveratrol in complex plant extract-the commercially available dietary supplement Regrapex-R-forte, which contains the extract of Vitis vinifera grape and extract of Polygonum cuspidatum root. Regrapex-R-forte is rich in stilbenes and other biologically active substances. Light microscopy imaging, confocal microscopy, and crystal violet staining were used to quantify and visualize the biofilm. The metabolic activity of biofilm-forming cells was studied by the tetrazolium salt assay. Amphotericin B had higher activity against planktonic cells; however, resveratrol and Regrapex-R-forte showed anti-biofilm effects, both in inhibition of biofilm formation and in the eradication of mature biofilm. The minimum biofilm eradicating concentration (MBEC80) for Regrapex-R-forte was found to be 2222 mg/L (in which resveratrol concentration is 200 mg/L). These methods demonstrated that Regrapex-R-forte can be employed as an anti-biofilm agent, as it has similar effect as amphotericin B (MBEC80 = 700 mg/L), which is routinely used in clinical practice.