In this study, we evaluated the clinical, biochemical, and pathological changes induced by oral administration of Acacia nilotica aqueous extract (ANAE) in broilers. A total of 71-day-old broilers were separated into one control group and six groups of 10 broiler chicks, which were subjected to a challenge. Various amounts of ANAE (1, 3, 5, 7.5, 10, and 15 g/kg b.wt) were given orally to broilers. Over a period of 10 days, indicators such as consumption of feed, alterations in body weight, and occurrences of poisoning or death were monitored. Blood samples were taken on day 5 after treatment for the purpose of evaluating biochemical parameters. Histopathological examination was performed on liver and kidney samples taken. The group that received ANAE at a dose of 15 g/kg showed decreased appetite, decreased locomotion, and increased hypersensitivity to touch, ultimately resulting in 100% mortality. The major lesions of histological liver tissue were cholestasis and adipose changes associated with significant increases in aspartate aminotransferase (AST), alanine aminotransferase (ALT), and alkaline phosphatase (ALP); Indicating liver impairment. Renal tissues exhibit marked inflammation, accompanied by glomerular atrophy and changes in urea levels. Based on the above data, Acacia nilotica is toxic to broilers only at 15 g/kg, demonstrating the safety profile of ANAE in broilers.