Cattle slaughterhouses generate wastewater that is rich in organic contaminant and nutrients, which is considered as high strength wastewater with a high potential for energy recovery. Work was undertaken to evaluate the efficiency of the 12 L laboratory scale conventional and a modified upflow anaerobic sludge blanket (UASB) reactors (conventional, R1 and modified, R2), for treatment of cattle slaughterhouse wastewater (CSWW) under mesophilic condition (35 ± 1 °C). Both reactors were acclimated with synthetic wastewater for 30 days, then continuous study with real CSWW proceeds. The reactors were subjected to the same loading condition of OLR, starting from 1.75, 3, 5 10, 14, and 16 g L−1d−1, corresponding to 3.5, 6, 10, 20, 28, and 32 g COD/L at constant hydraulic retention time (HRT) of 24 h. The performance of the R1 reactor drastically dropped at OLR 10 g L−1d−1, and this significantly affected the subsequent stages. The steady-state performance of the R2 reactor under the same loading condition as the R1 reactor revealed a high COD removal efficiency of 94% and biogas and methane productions were 27 L/d and 89%. The SMP was 0.21 LCH4/gCOD added, whereas the NH3-N alkalinity ratio stood at 651 mg/L and 0.2. SEM showed that the R2 reactor was dominated by Methanosarcina bacterial species, while the R1 reactor revealed a disturb sludge with insufficient microbial biomass.