Lignocellulosic biomass from the agricultural sector including crop residues and animal manure is an abundantly available resource in Pakistan. Anaerobic digestion (AD) can valorize this biomass to produce much-needed bioenergy in the form of biogas as well as the biofertilizer for crop production. However, the anaerobic microorganisms cannot efficiently degrade the lignin resulting in poor biogas yield and quality. A suitable pretreatment to degrade biomass lignin and the use of co-substrates with cattle manure during AD, to balance the nutrients, can improve both the biogas yield and quality. Therefore, the objectives of the proposed project are to 1. determine the potential of microaerobic pretreatment to degrade lignin of the biomass, and 2. subsequently establish the biogas potential of pre-treated agro-industrial co-substrates and animal manure both at laboratory and pilot scale. The metagenomic analysis will be performed to study the shift in microbial ecology during aerobic and anaerobic treatment of biomass. Since the biogas plants already operational in the community use buffalo manure, therefore, the primary substrate for this research will be buffalo manure. The co-substrates used in this research will be poultry manure, wheat straw, corn cobs and stalks, rice husk, and sugarcane bagasse. The co-substrates are selected because of their availability in Pakistan and suitability for the AD process. The results of this research will help in establishing the biogas/biomethane potential of locally available organic substrates, measures to improve the performance of currently operational biogas plants and develop guidelines for future biogas development initiatives in Pakistan.