An originally isolated baculovirus, Spodoptera litura multiple nucleopolyhedrovirus (SpltMNPV) was serially passed through the S. litura larvae for upto four generations to determine the mean number of occlusion bodies (OBs) harvested per larva and their efficacy in terms of infectivity, feeding cessation and speed of kill of host larvae. The results revealed that the mean number of OBs harvested per larva increased significantly with increase in the dose of SpltMNPV at each passage and the yield was significantly lower in original stock wild-type SpltMNPV (P0) as compared to serially passed SpltMNPV (P1, P2, P3 and P4). Laboratory bioassays indicate that median lethal doses (LD50), median times to feeding cessation (FT50) and median survival times (ST50) of P0, P1, P2, P3 and P4 were significantly different from each other. The OBs of each passage when tested for their cross-infectivity against Spodoptera exigua and Spilarctia obliqua revealed significant reduction in their mortality. These results indicate that serially passed SpltMNPV is more host specific and more effective biocontrol agent than the original stock wild-type virus and can be adopted for mass production as a viral pesticide for control of the S. litura.