Fighting resistance to antibiotics and chemotherapeutics has brought bioactive peptides to the fore. Peptaibols are short α-aminoisobutyric acid-containing peptides produced by Trichoderma species. Here, we studied the production of peptaibols by Trichoderma atroviride O1 and evaluated their antibacterial and anticancer activity against drug-sensitive and multidrug-resistant bacterium and cancer cell lines. This was substantiated by an analysis of the activity of the peptaibol synthetase-encoding gene. Atroviridins, 20-residue peptaibols were detected using MALDI-TOF mass spectrometry. Gram-positive bacteria were susceptible to peptaibol-containing extracts of T. atroviride O1. A synergic effect of extract constituents was possible, and the biolo-gical activity of extracts was pronounced in/after the peak of peptaibol synthetase activity. The growth of methicillin-resistant Staphylococcus aureus was reduced to just under 10% compared to the control. The effect of peptaibol-containing extracts was strongly modulated by the lipoteichoic acid and only slightly by the horse blood serum present in the cultivation medium. Peptaibol-containing extracts affected the proliferation of human breast cancer and human ovarian cancer cell lines in a 2D model, including the multidrug-resistant sublines. The peptaibols influenced the size and compactness of the cell lines in a 3D model. Our findings indicate the molecular basis of peptaibol production in T. atroviride O1 and the potential of its peptaibol-containing extracts as antimicrobial/anticancer agents.

Understanding the interplay between antifungal resistance, stress adaptation, and virulence in Aspergillus fumigatus is critical for more effective treatment outcomes. In this study, we investigate six clinical isolates of A. fumigatus from the hospitals of the Czech Republic, focusing on their resistance profiles, stress responses, and survival mechanisms under antifungal pressure. Notably, we have shown that azole-susceptible strains were able to form persister cells under supra-MIC concentrations, highlighting an emerging non-genetic survival mechanism. Stress response profiling demonstrated differential susceptibility to agents targeting signal transduction pathways, as principal component analysis proved that even azole-resistant strains might rely on these pathways. Combinatorial treatment with posaconazole and dithiothreitol enhanced antifungal efficacy regardless of the susceptibility of the strains. Fitness assays revealed that azole resistance imposed a competitive disadvantage in azole-free conditions. In vivo virulence assessment in Galleria mellonella larvae revealed strain-specific pathogenicity that did not directly correlate with resistance. Together, our findings illustrate the multifactorial nature of fungal survival and emphasize that stress adaptation, tolerance, and persistence significantly affect treatment efficacy and outcomes, even in the absence of classical resistance mechanisms. Targeting stress response pathways emerges as a promising strategy to enhance the efficacy of existing antifungal agents and manage resistance in A. fumigatus.

This work was focused on the characterization of novel isolate of Trichoderma atroviride O1, found in the forest around the village of Zázrivá (the Northern Slovakia, region Orava). The isolate was identified by sequencing its internal transcribed spacer (ITS) region of rDNA. T. atroviride O1 stimulated the development of lateral roots of model plant Lepidium sativum . Simultaneously, the isolate has proved its high mycoparasitic potential as it displayed the ability to attack colonies of phytopathogenic fungi ( Alternaria alternata, Fusarium culmorum, Botrytis cinerea ). This isolate produced secondary metabolites, which were isolated and tested for the antimicrobial activity against gram-positive bacteria Staphylococcus epidermidis and Staphylococcus aureus. The growth of these bacteria was suppressed to 10 % and 40 %, respectively. The suppression of the growth of two Candida species was also strong (10 % growth). However, growth parameters of three phytopathogenic fungi (Alternaria alternata, Botrytis cinerea and Fusarium culmorum ) were less affected (75 % growth in comparison with the control). Attempts were made to characterize secondary metabolites isolated from T. atroviride O1. Known peptaibols, 20—21 amino acid long, but also shorter peptides, were detected by MALDI-TOF mass spectrometry. Thus, this study demonstrates the plant growth promotion, strong mycoparasitic potential and antimicrobial activity of the isolate T. atroviride O1, which could be in part ascribed to the production of secondary metabolites. This isolate does have a potential in the biocontrol in eco-farming. Further study, particularly, the identification of produced secondary metabolites, is needed.