Enhancement of Biomethane Yield from Spent Mushroom Substrate: Biological Pretreatment with the Chlamydospores of Trichoderma viride

Fungal chlamydospores are asexual spores formed by fungi under adverse conditions and could be used in biological pretreatment for biogas projects fed by lignocellulosic substrates. In this study, Trichoderma viride (Tv) chlamydospores were used as the pretreatment agent to enhance the methane yield of spent mushroom substrates (SMSs). Lignocellulosic composition, methanogenesis performance, and anaerobic microbial communities were investigated for different Tv pretreatment durations (0 h, 12 h, 24 h, 48 h, 96 h, and 192 h). The results showed that the optimal Tv pretreatment duration was 24 h, and the cumulative methane yield reached 173.4 mL/gVS, which was 16.8% higher than that of the control. A pretreatment duration longer than 48 h was not conducive to methanogenesis. Sequencing analysis of anaerobic microbial communities showed that the pretreatment duration was directly proportional to the relative abundance of Tv at the beginning of digestion. When the initial Tv abundance was higher than 50%, Trichoderma became the absolute dominant fungus with an abundance higher than 97% in fungal communities in the later stage of digestion. The correlation network among fungi, bacteria, and archaea showed that Tv was directly related to 11 genera, and through these taxa, Tv affected 58% of the taxa in the whole microbial network. Cost accounting showed that Tv pretreatment has a net income of 45.5 CNY/1000 kg SMS, and is a promising technology. This study provides important guidance for the use of fungal chlamydospores in pretreatment and also promotes the understanding of fungi in anaerobic digestion.