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Biological pre-treatment of lignocellulosic biomass has an advantage of low chemical and energy use. The feasibility of
biological pretreatment of wheat straw with fungal co-cultures of Trichoderma viride, Aspergillus niger and Fusarium
oxysporum for efficient biogas and methane production was investigated in the present study. Firstly, physicochemical
characteristics of wheat straw in addition to analysis of cow dung, well known to contain methanogenic consortia. Pretreated
wheat straw with the investigated fungal co-cultures for 7 days was conducted. Changes in TOC%, TKN%, C/N, pH, TDS, TP
and TK further demonstrated that fungal pretreatment was effective. Subsequently, cumulative biogas production 51.8 l/KgVS
compared to untreated substrate which produced 35.7 l/KgVS. Moreover, the maximum methane production was found to
be 28.01 l/KgVS which was 0.9 l/KgVS in the wheat straw without pre-treatment. By using VIT�® gene probe technology, the
methanogenic bacteria were identified as members of the genus Methanosarcina. The present study suggested the potential role
of using the investigated fungal co-cultures for wheat straw crop residues pre-treatment for high-yield methane and biogas.
In this sense, the use of cheap digesting microflora starter for biofuel production might reduce the high cost of other pretreatment
methods.