Chlorpyrifos (CPF) is one of the popular crop pests that is widely used in the agriculture sector. In the aquaculture sector, several strategies were applied to mitigate the negative impacts of CPF on aquatic animals through the application of feed additives. Spirulina platensis (SP) is well reported as a functional feed additive to enhance the physiological, immunological, and anti-oxidative status in farmed fish. In the current study, fish were randomly stocked in five groups where the first and second groups reared in clean water without CPF toxicity and were fed the basal diet without SP or with SP at 1%, respectively. Meanwhile, the third, fourth, and fifth groups were exposed to CPF (15 μg/L) in rearing water and fed SP at 0, 0.5, and 1%, respectively. Under CPF exposure, SP displayed preventive and restorative impacts against toxicity in Nile tilapia. Fish fed SP-supplemented diet showed decreased alanine aminotransferase (ALT), aspartate aminotransferase (AST), alkaline phosphatase (ALP), cholesterol, urea, and creatinine as well as increased total protein, albumin, superoxide dismutase (SOD), and catalase (CAT) activities. The outcomes suggested that SP is efficient in protecting Nile tilapia from CPF toxicity by increasing the antioxidative response.