Envenomation by snakes of the genus Bothrops is characterized by local pain, systemic bleeding, tachycardia, acute renal failure, as well as swelling and redness caused by the enzymatic and proteolytic action of venom. The toxicological and histological effects of the venom of Bothrops itapetiningae, one of the smallest pit vipers in Brazil, were characterized using BALB/c male mice as models. Venom impacts on liver, kidney, and plasma enzymes that were evaluated at six concentrations. We observed lethality, toxicity, edema forming, and hemorrhagic symptoms such as polymorphonuclear infiltration, hydropic degeneration, and ischemic necrosis. Moreover, increases of alanine aminotransferase, aspartate aminotransferase, alkaline phosphatise, and creatinine levels were observed in the presence of venom. We assessed LD50, minimal hemorrhagic dose, minimal edematogenic activity, and platelet aggregation, improving our knowledge of B. itapetiningae venom toxic activity.