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A study was carried out in order to investigate the influence of Moringa oleifera leaf extract on reducing of methane production in sheep. The experiment by in vitro methode was done from June 2018 until October 2018. The treatments were ration with addition of three level of Moringa oleifera leaf extract of 0%, 0.25%, and 0.50% from dry matter (DM) of diet, respectively. Moringa oleifera leaves are dried in an oven at 60ºC for 2 x 24 hours, then ground to make extracted using ethanol. A Completely Randomized Design with six replications was applied in this experiment. The Rumen fluid was obtained from three thin-tailed sheep and was used as a source of inoculum. The diet consists of concentrate and ammoniated rice straw ratio of 60:40 based on DM and the concentrate consists of two parts of rice bran and one part of coconut meal. The results of the variance analysis and the orthogonal polynomial test indicated that the level of 0.5% Moringa oleifera leaf extract lowest of the number of protozoa and methane production, and the highest number of bacteria and microbial protein synthesis.