Babesia bigemina is the cause of bovine babesiosis. B. bigemina-infected red blood cells (iRBCs) were passaged in vitro for the attenuation. Methods: We cultured Pakistani isolate of the parasite in three different culture media. Whole blood was collected from splenectomized and intact crossbred young calves. The parameters included were hematological profile, catalase activity, osmotic fragility and lipid profile. Cell culture media (M-199, RPMI 1640 and DMEM) were used to find out the

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... y of parasites iRBCs.Result: Highest PPE level was found up to 6.0% on 72 h post-culture in M-199 medium. Furthermore, no significant difference in catalase activity while significant difference in osmotic fragility were observed. However, lipid profile was significantly less in infected animals except in Babesia-infected. M-199 was the most appropriate medium for the in vitro cultivation of B. bigemina. Our findings would help us for in vitro cultivation of babesia-infected RBCs for its attenuation.