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The present study was designed to assess the invivo effect of stem bark methanol extract of Acacia nilotica in 40 wistar rats infected with Trypanosoma brucei brucei. The rats weighing abouthe aim of this study was to determine the invivo effect of stem bark methanol extract fractions of Acacia nilotica in 40 wistar rats infected with Trypanosoma brucei brucei. The rats weighing about 110grams were randomly allotted to different groups of 4 rats each. The groups were treated with 300, 400 and 500mg/kg doses of each of the fractions. The methanol extract was further fractionated into different fractions using 100% ethyl acetate, 100% n- butanol, 100% n- hexane and water. The yield shows ethyl acetate with highest percentage of fraction followed by butanol. N-hexane with the lowest percentage. Three (3) days pre patent period was observed after inoculating of the parasite. Parasiteamia was monitored daily while Parked Cell Volume (PCV) was determined at one-day interval during the course of infection. The 100% ethyl acetate and 100% n- butanol fractions showed antitrypanosomal activities against T. b. b. in infected experimental animals. The Butanol fraction (500mg/kg dose) cured the rats after 5 days’ treatment period while Ethyl acetate fraction at 400mg/kg and 500mg/kg doses cleared the parasite after 6 days’ treatment. Water and n-hexane fractions shows little or no effects against Trypanosoma brucei brucei in experimental animals. The study concluded that butanol and ethyl acetate faction contains active ingredients with therapeutic potentials against trypanosomiasis. Keywords: Trypanosoma brucei brucei, Acacia nilotica, Stem bark, anti-trypanosomal.