Diaphanosoma excisum, the fresh water cladocera was cultured with different concentrations of cow urine distillate (CUD) to evaluate its growth performance, nutritional composition and the antibacterial potential of hemolymph. The D.excisum showed a dose dependent increase in growth and concentration of biomolecules with the treatment of CUD. The LC50 concentration of CUD against D.excisum was 0.683 μg/ml. The hemolymph isolated from the D.excisum culture with CUD showed effective antibacterial activity. The study thus suggests that CUD could be utilized for the enhanced of growth and nutritional composition of D.excisum under optimized growth conditions and thus can be mass cultured to provide the live food in aquaculture.