This study evaluated the effect of dietary inclusions of Garcinia kola (GK) seed on geotactic behavior and some oxidative stress markers in wildߚtype fruit flies (Drosophila melanogaster). Flies were raised on diet supplement with GK seed for 5 days. The negative geotactic behavior of flies which was used to evaluate their locomotor performance was thereafter evaluated. The flies were subsequently homogenized and the reactive oxygen species (ROS) level, acetylcholinesterase (AChE), catalase and glutathioneߚSߚtransferase (GST) activities, as well as nitric oxide (NO) and total thiol contents were assayed. The phytochemical constituents of GK seed were also determined. It was observed that higher dietary inclusions of GK seed reduced the survival rate of D. melanogaster more significantly compared to control flies. Also, higher dietary inclusions of GK seed significantly reduced locomotor performance and AChE activity, while the ROS level was increased compared to the control. Activities of GST and catalase were significantly increased in flies fed diet supplemented with higher GK seed inclusions but their NO content was significantly reduced compared to control. Phytochemical analysis of GK seed revealed abundance of saponin > glycosides > alkaloids > phenols > flavonoids. These results have shown that dietary inclusion of GK seed at higher concentrations reduced survival rate of D. melanogaster and impaired cholinergic system, with elevated activities of some antioxidant enzymes under acute exposure. These observations could be associated with bioactivities of predominant phytochemicals in GK seed such as saponin and glycosides which have been reportedly toxic at high concentration. Therefore, this study suggests that high consumption of GK seed could induce some toxicological effects and moderate consumption is hence recommended.