Energy production and recycling from chicken manure (CM) is a sustainable and environmentally friendly approach. The CM is considered a valuable bioresource for methane production. The commercial applications of colimoxin for the poultry bird health; its traces are being ejected in CM and it becomes toxic for the methanogenic archaea during the anaerobic digestion process. The influences of the colimoxin were investigated on the anaerobic co-digestion performance of the CM and wheat straw (WS). The CM is co-digested with WS at carbon to nitrogen ratio of 25 to reduce the nitrogen contents of the CM. To optimize the colimoxin dose for methane generation; eight different colimoxin treatments were applied with varying colimoxin dose. The lower colimoxin dose was found healthy for the microbes and Treatment-4 resulted in 23% methane enhancement, while the higher colimoxin doses inhibited the methane production. The synergistic influences of the colimoxin dose on the volatile fatty acids generation and utilization were deeply monitored. The regression models were developed by using second degree polynomial regression models to optimize the colimoxin dose for the commercial biogas facilities and for the commercial poultry control sheds. The onsite methane generation would result in in-situ electrical energy production for the rural community.