Aqueous extract of antidiarrhoeal formulation (ADF) containing Holarrhena antidysentrica, Aeglemarmelos and Punica granatum was investigated for antidiarrhoeal activity against charcoal-induced gut transit, serotonin-induced diarrhoea and PGE2-induced small intestine enteropoolingin rats. The control, standard and test groups of experimental animals were administered withnormal saline (p.o.), diphenoxylate hydrochloride (5 mg/kg, p.o.) and ADF (250 mg and 500 mg/kg, p.o.) respectively except the control group of PGE2-induced small intestine enteropoolingwhich received only 5% ethanol in normal saline (i.p.). Charcoal (10 ml/kg, p.o.) and serotonin(600 µg/kg, i.p.) were administered after 30 min, while PGE2 (100 µg/kg, p.o.) was administeredimmediately afterwards. The distance traveled by charcoal in small intestine was measured after15 and 30 min of charcoal administration, diarrhoea was observed every 30 min for six hour afterserotonin administration and the volume of intestinal fluid was measured after 30 min of PGE2administration. Oral administration of ADF significantly inhibited the frequency of defaecationand decreased the propulsion of charcoal meal through the gastrointestinal tract, reduced thewetness of faecal dropping in serotonin-induced diarrhoea and also reduced the PGE2-inducedsmall intestine enteropooling. ADF may have potential to reduce the diarrhoea in rats.