The present study deals with the impact of choline chloride supplementation, applied directly into the pond water in a semi-intensive farm culture system for 90 d on two Indian Major Carps (IMCs) (Catla catla, Labeo rohita) and two air-breathing teleostean fishes (Anabas testudineus, Clarias batracus).The experiment was conducted in two seasons (breeding and dry) under choline (treated) and non-choline (control) exposure. Biochemical parameters, viz., PRO (total protein) content gained maximum significantly (p< 0.01) in treated-breeding with the lowest in control-dry in intestine, liver, muscle, brain. IMCs showed significant (p< 0.01) increased amylase activity in the liver and intestine in treated-dry and treatment-breeding conditions, while liver amylase activity in both the seasons increased manifolds significantly (p< 0.01) in air-breathing fishes, showing a sharp decline only in the intestine. Protease and lipase activity in the intestine and liver of choline-fed fishes disclosed declining trend in the breeding season significantly (p< 0.01), but treated (choline-fed) IMCs under dry revealed the significant (p< 0.01) increasing and decreasing trend of protease and lipase activity respectively; the reverse trend at dry season was found in air-breathing fishes under choline-fed condition. ALP (alkaline phosphatase) in intestine indicated significant (p< 0.01) lowest activity in treatment-breeding, highest in treatment-dry. This study explored the higher activity towards total protein generation, especially muscle; higher carbohydrate digestion; elevated lipid digestion in choline-fed fishes. So, the fishes from this farm-based pisciculture system are healthy and its yield can be prescribed as a rich source of quality fish food for human health.