This research article published by AIMS Press, 2020The performance of experimental batch-reactor loaded with slaughterhouse waste at mesophilic temperature was investigated as well as the inhibition of both ammonia and sulfide concentration in the aqueous phase. The digester was operated for 68 days by evaluating the process stability basing on controlling parameters such as pH, volatile fatty acids and alkalinity in relation to the methane produced. The maximum CH4 content of 69.6% was achieved at 0.37 VFA/Alkalinity ratio and pH of 7.51 during day 37 of anaerobic digestion. However, a sudden increase of ammonia nitrogen in the digester from day 44 to day 68 decreased the methane content about 62.15% from 65% to 24.6%. Similarly, as the amount of sulfide content decreased in the liquid phase, gaseous H2S was elevated up to 132 ppm in the 68th day. During this time, it was observed that the ratio of VFA/Alkalinity decreased to 0.16, with a very low concentration of VFA, which was 150.92 mg/L. This phenomenon indicated that all the acids produced were consumed by methanogens and ammonia inhibition was at the highest rate due to the increase of ammonia nitrogen concentration in the last days of digestion. Furthermore, among of peculiar characteristic shown by slaughterhouse waste is the ability to maintain the pH above 7 without the addition of any buffering agent throughout the AD process. Meanwhile, the evaluation of the level of both ammonia and sulfide in the aqueous phase revealed that the inhibitory effect of ammonia concentration was higher than sulfide concentration.