Surface properties of mycoparasitic Pythium species and their interaction with model materials.

Pythium oligandrum, a soil-born oomycete, is an effective biological control agent exhibiting antagonistic and parasitic activity against pathogenic fungi. This study is the first attempt to characterize its surface properties and to apply models of physicochemical interactions (thermodynamic, DLVO and XDLVO) to quantify its adhesion properties to a model material, represented by magnetic beads (MB). The predictions of interaction models were based on experimental data (contact angles, zeta potentials, size). Adhesion intensities (AI) were determined experimentally taking advantage of MB with different surface properties. The role of weak physicochemical interactions was estimated by comparing experimental AI with model predictions. The results revealed that the surface properties of the three Pythium spp. studied were very similar and fell within the range for hydrophilic microorganisms (ΔGTOT > 0) with a predominantly negative surface charge. The most reliable description of AI was obtained using the DLVO model, including Lifshitz-van der Waals and electrostatic interactions. The highest AI between Pythium spp. and all three MB was observed at pH 3, which was supported by the DLVO prediction. The greater agreement between the sphere-sphere geometric version of the DLVO model and experiment suggests that the surface protrusions of the oospores increase the efficiency of adhesion. The surface properties of the pathogenic fungi, characterized in this work, fell within the range defined by MB and therefore it can be expected that their physicochemical interactions with Pythium spp. will also be favourable.

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.

Exploring Non-Thermal Plasma and UV Radiation as Biofilm Control Strategies against Foodborne Filamentous Fungal Contaminants.

In recent years, non-thermal plasma (NTP) has emerged as a promising tool for decontamination and disinfection within the food industry. Given the increasing resistance of microbial biofilms to conventional disinfectants and their adverse environmental effects, this method has significant potential for eliminating biofilm formation or mitigating the metabolic activity of grown biofilms. A comparative study was conducted evaluating the efficacy of UV radiation and NTP in eradicating mature biofilms of four common foodborne filamentous fungal contaminants: Alternaria alternata, Aspergillus niger, Fusarium culmorum, and Fusarium graminearum. The findings reveal that while UV radiation exhibits variable efficacy depending on the duration of exposure and fungal species, NTP induces substantial morphological alterations in biofilms, disrupting hyphae, and reducing extracellular polymeric substance production, particularly in A. alternata and F. culmorum. Notably, scanning electron microscopy analysis demonstrates significant disruption of the hyphae in NTP-treated biofilms, indicating its ability to penetrate the biofilm matrix, which is a promising outcome for biofilm eradication strategies. The use of NTP could offer a more environmentally friendly and potentially more effective alternative to traditional disinfection methods.

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.

Taxonomic analysis reveals host preference of rare fungi in endophytes of Vitis vinifera from the Czech Republic.

This paper represents the results of screening a diversity of fungal endophytes associated with Vitis vinifera leaves and canes in the Czech Republic. The characterization of strains is based on morphological and phylogenetic analyses of ITS, EF1α and TUB2 sequence data. Our strain selection covers 16 species and seven orders belonging to Ascomycota and Basidiomycota. Together with ubiquitous fungi, we report on several poorly known plant-associated fungi, Angustimassarina quercicola (= A. coryli, a synonym proposed in this study) and Pleurophoma pleurospora. Other species, such as Didymella negriana, D. variabilis, Neosetophoma sp. (species identical or sister to N. rosae), Phragmocamarosporium qujingensis and Sporocadus rosigena, have so far been little known and rarely found, but are frequent on V. vinifera in different parts of the world and obviously belong to a microbiota with a strong preference for this plant. Detailed taxonomical identification allowed us to identify species with apparent stable associations with V. vinifera, for which further interactions with V. vinifera can be expected. Our study is the first to focus on V. vinifera endophytes in Central Europe and expands the knowledge about their taxonomy, ecology and geography.

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.

Boswellic Acids as Effective Antibacterial Antibiofilm Agents.

Boswellic acids are biologically active pentacyclic terpenoid compounds derived from Boswellia sp. plants. Extracts containing these acids have a number of positive effects on human health, especially in the treatment of inflammation, arthritis, or asthma. With increasing resistance to common antibiotics, boswellic acid-containing extracts could serve as an alternative or work in synergy with commonly available preparations. This study aims to determine the effect of boswellic acids on suspension cells and biofilms of Staphylococcus epidermidis, Enterococcus faecalis, and Escherichia coli. The antimicrobial and antibiofilm effect found was compared with commonly available antibiotics to control these undesirable microorganisms. The synergistic effect of boswellic acids and common antibiotics on the growth of these microorganisms was also determined. All tested microorganisms showed a positive additive effect of antibiotics and boswellic acid extract. The most significant effect was found in Enterococcus faecalis ATCC 29212 in a combination of 0.2 × MIC80 erythromycin (0.2 mg/L) and 0.8 × MIC80 boswellic acid extract (16 mg/L).

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.