Bitumen rejuvenators are used to improve or restore the physical and mechanical performance of aged bitumen. Traditional bitumen rejuvenators are a product of crude oil. As crude oil production declines and the environmental and financial costs of crude oil increase, there is an urgent need to identify more environmentally sustainable bitumen rejuvenator alternatives. Bitumen rejuvenators generated from biological sources offer an environmentally friendly and economically viable alternative to the crude oil-based bitumen rejuvenators. This paper describes a study wherein microalgae oil was used as an aged bitumen rejuvenator. The chemical, physical, and mechanical effects of microalgae oil on 70/100pen aged bitumen were investigated. The results indicate that microalgae oil has the potential to be used as an aged bitumen rejuvenator.