Morris Water Maze

The Morris Water maze test (MWM) was performed with a slightly modified protocol.^36,37 The MWM equipment comprised a circular water pool (80 cm in diameter with a 40 cm high wall) that was partitioned into four quadrants ie, Q1, Q2, Q3, and Q4 with external visual signs on the laboratory walls for navigation. The pool was filled with water to a depth of 30 cm and milk whitener was added to make the water opaque and it was maintained at a temperature of 26.0 ± 1.0 ℃. A platform, 6 cm in diameter was positioned in the center of Q3 and it was maintained in position throughout the test activity. MWM was used to estimate the effect of ASEE and ASB extracts of Acacia stenophylla bark on changes in spatial memory. On the first day of the protocol, each mouse was allowed 60 seconds in the pool to swim and explore and then transferred to the home cage. The animals were then administered test extracts daily for five consecutive days. Thirty minutes after dosing with bark extract, scopolamine or diazepam were injected in all but the control group. Thirty minutes later, each animal underwent two consecutive platform location trials of 120s with an interval of 30 minutes on four consecutive days. The mouse platform location time or escape latency (EL) was noted. Animals were then permitted to reside on the platform for the 20s and then transferred to the home cage. Animal failure to locate the platform during the 120s trial meant that it was guided manually to the platform and then transferred to the home cage. On the fifth day, each mouse underwent a 120s probe trial in the absence of the platform and after administration of bark extract and/or scopolamine/diazepam. This determined the time each animal spent in the correct quadrant (Q3) as an indication of memory retention.