In the present study the detailed histochemical localization of alkaline phosphatase at the tissue level of Cotylophoron cotylophorum, a stomach fluke of cattle, has been described. Cotylophoron cotylophorum were collected from the rumen of cattle in local abattoirs. Frozen sections were cut and histochemical localization of Alkaline phosphatase was done by using Calcium Cobalt method [17] in which alkaline phosphatase deposits appear black or brownish black. In the present study substantial amount of alkaline phosphatase was observed in the various structures of Cotylophoron cotylophorum. Intense reaction for alkaline phosphatase was observed in intestinal caeca, excretory vesicle, tunica of testes and ovary. Moderate reaction for alkaline phosphatase was observed in oral sucker, acetabulam, vitellaria, tegument and sub-tegument of Cotylophoron cotylophorum. Parenchyma showed weak reaction for alkaline phosphatase in the present study. The possible functional role of alkaline phosphatase in different tissues has been discussed.