Removal of the cytostatic drugs bleomycin and vincristine by white-rot fungi under various conditions, and determination of enzymes involved, degradation by-products, and toxicity

Anticancer drugs show recalcitrance to conventional wastewater treatments; thus, they are present in aquatic systems and pose an environmental threat. Fungi represent a promising biological alternative for wastewater treatments. Therefore, the goals of this work were to assess the potential of white-rot fungi (Fomes fomentarius (CB13), Hypholoma fasciculare (CB15), Phyllotopsis nidulans (CB14), Pleurotus ostreatus (BWPH), and Trametes versicolor (CB8)) for removing bleomycin and vincristine, and to investigate the impacts of various conditions (shaking, aeration, or biomass immobilization) on the process. The removal capacities were measured using Ultra-Performance Liquid Chromatography (UPLC) coupled with Mass Spectrometry (MS) and preceded by Solid Phase Extraction (SPE). We further identified major drugs degradation products; determined the fungi's main enzyme activity profiles (laccase, manganese and lignin peroxidases); and examined the toxicities of post-processed samples against Lemna minor, Daphnia magna and Pseudomonas putida. In just 2 days, all strains (except for P. nidulans) removed >90 % of vincristine, nearly completely eliminating the drug over time. Bleomycin content reduction occurred with T. versicolor or H. fasciculare, respectively reaching 55 % and 83 % drug elimination after 9 days. Oxygen was found to be crucial for cytostatics degradation, with their highest removal rates occurring in samples with air supply (aeration or agitation). Laccase was the only tested enzyme associated with cytostatics elimination. Drug biodegradation was followed by detoxification, demonstrating the utility of fungi in cytostatics removal.