A farm-scale biogas desulfurization system was designed and tested for H2S removal efficiency from livestock biogas. This work assesses the H2S removal efficiency of a novel farm-scale biogas bio-desulfurization system (BBS) operated for 350 days on a 1,000-head pig farm. Experimental data demonstrated that suitable humidity and temperature can help sulfur-oxidizing bacteria to form active bio-films on the bio-carriers. The daily average removal rate increased to 879.16 from 337.75 g-H2S/d with an average inlet H2S concentration of 4,691 ± 1,532 mg/m3 in biogas. Thus, the overall (0–350 days) average H2S removal efficiency exceeded 93%. The proposed BBS overcomes limitations of H2S in biogas when utilizing pig farm biogas for power generation and other applications.