Biogas plants have been started to expand recently in Greece and their positive contribution to the economy is evident. A typical case study is presented which focuses on the long-term monitoring (lasting for one year) of a 500 kW mesophilic biogas plant consisting of an one-stage digester. The main feedstock used was cow manure, supplemented occasionally with chicken manure, corn silage, wheat/ray silage, glycerine, cheese whey, molasses and olive mill wastewater. The mixture of the feedstocks was adjusted based on their availability, cost and biochemical methane potential. The organic loading rate (OLR) varied at 3.42 ± 0.23 kg COD m−3 day−1 (or 2.74 ± 0.18 kg VS m−3 day−1) and resulted in a stable performance in terms of specific biogas production rate (1.27 ± 0.12 m3 m−3 day−1), biogas yield (0.46 ± 0.05 m3 kg−1 VS, 55 ± 1.3% in methane) and electricity production rate (12687 ± 1140 kWh day−1). There were no problems of foaming, nor was there a need for trace metal addition. The digestate was used by the neighboring farmers who observed an improvement in their crop yield. The profit estimates per feedstock indicate that chicken manure is superior to the other feedstocks, while molasses, silages and glycerin result in less profit due to the long distance of the biogas plant from their production source. Finally, the greenhouse gas emissions due to the digestate storage in the open air seem to be minor (0.81% of the methane consumed).