The data are inconsistent about the ability of dietary omega-3 fatty acids to prevent age-associated cognitive decline. Indeed, most clinical trials have failed to demonstrate a protective effect of omega-3 fatty acids against cognitive decline, and methodological issues are still under debate. In contrast to human studies, experiments performed in adult rodents clearly indicate that omega-3 fatty acids supplement can improve behavioural and cognitive functions. The inconsistent observations between human and rodent studies highlight the importance of the use of non-human primate models. The aim of the present study was to address the impact of omega-3 fatty acids (given in the form of dietary fish oil) on exploratory activity, emotional status and spatial reference memory in the aged mouse lemur, a non-human primate.Aged animals fed fish oil exhibited decreased exploratory activity, as manifested by an increase in the latency to move and a reduced distance travelled in an open-field. The fish oil-supplemented animals exhibited no change in the anxiety level, but they were more reactive to go into the dark arms of a light/dark plus-maze. In addition, we found that fish oil supplementation did not significantly improve the spatial memory performance in the Barnes maze task.This study demonstrated for the first time that a fish oil diet initiated late in life specifically modifies the exploratory behaviour without improving the spatial memory of aged non-human primates. Omega-3 fatty acid supplementation may be effective when started early in life but less effective when started at later ages.