Paramphistomosis is one of the major pathogenic diseases in domestic animals and responsible for heavy economic loss in terms of reduced milk, meat and wool production. Cotylophoron cotylophorum is more prevalent in Tamilnadu. In the present investigation, the effect of Syzygium aromaticum ethanol extract on the enzymes of carbohydrate metabolism viz. pyruvate kinase, phosphoenolpyruvate carboxykinase, lactate dehydrogenase, malate dehydrogenase, fumarate reductase and succinate dehydrogenase of Cotylophoron cotylophorum was studied in vitro. The parasites were incubated in five different sub-lethal concentrations of ethanol extract of Syzygium aromaticum viz. 0.005, 0.01, 0.05, 0.1 and 0.5mg/ml for 2, 4 and 8 h. The activity of the enzymes was assayed using standard procedures. The enzyme activity was expressed in terms of protein. The data obtained were analyzed statistically. Ethanol extract of Syzygium aromaticum significantly inhibited the enzymes of carbohydrate metabolism and the percentage of inhibition was dose and time dependent. Inhibition of these enzymes leads to decreased ATP production which may be fatal to the parasites. The present study validates the anthelmintic property of ethanol extract of Syzygium aromaticum against C. cotylophorum.