The effectiveness of an enzymatic cocktail, collagenase and trypsin, on the absorptive function of ileal segment was studied to evaluate the changes within groups after performing ILEOCYSTOPLASTY in an animal model. Twenty-one female mixed-bred Persian dogs were randomized into 7 groups. The groups included: 1) negative control group (NCG) in which no ILEOCYSTOPLASTY was performed (n = 3); 2) positive control group (PCG) in which routine ILEOCYSTOPLASTY was performed (n = 3) and groups 3 to 7) those underwent ILEOCYSTOPLASTY with the ileal segment being treated with collagenase and trypsin for 5, 10, 15, 20 or 25 min (groups, 5 minutes enzymatic treatment group (5METG), 10METG, 15METG, 20METG and 25METG; each containing three dogs). To perform the glucose absorption test, 50% dextrose was instilled into the bladder five weeks after surgery. The blood glucose level (BGL) was measured in each group before the study (T1), two weeks after the surgery in PCG and other treatment groups (T2), before anaesthesia (T3), after anaesthesia at 5-minute intervals up to 25 min (T4 to T9, respectively). There was no significant difference in BGL in T1 to T3 and T4 to T9 in 25METG and NCG, however, BGL was found significantly higher in T1 to T3 and T4 to T9 in PCG, 5METG, 10METG, 15METG and 20METG. It can be concluded that collagenase and trypsin can affect absorptive function of the neobladder and are able to reduce the absorptive function, particularly in early days after the surgery. Moreover, these results also confirm that 50% dextrose instilled into the neobladder and native urinary bladder will not increase BGL in 25METG and NCG even with increasing the time of exposure of neo- or native urinary bladder to enzymatic cocktail till 25 min.