The document provides protocols for experiments involving rats using a running wheel over 3 weeks and a Morris water maze over 2 additional weeks. For the running wheel experiment, rats will individually spend 10 minutes in the wheel with their laps recorded, completing the task in around 124 minutes total. The water maze experiment involves placing rats in a pool to locate a hidden platform within 60 seconds, recording their times over 6 trials each, taking around 143 minutes in total to complete.

1. Protocol for weeks 1-3: Aerobic Intensity With Running Wheel
Total experiment time given:160 min
Materials:
● Running wheel
● clipboard
● pens or pencils
● Transfer cage (1) with lid
● stopwatch
● observation sheets
Set-up (Will take 10 min on day of experiment)
● Obtain running wheel from the other room.
● Set up the wheel next to the transfer cages for easy transfer.
● Place observation sheets in clipboard and have pens and pencils ready.
Instructions:
1. Assign one group member to be rat handler/transferer, another to record the number of
laps taken, another to record rat observations (e.g., levels of stress, anxiety, etc.) Other
group members can help with rat organization (keeping track of which rat is up next),
note taking on observations, and keeping time.
2. Administer the independent variables, water and water+supplement. Wait 10 minutes to
allow rats to digest the substance given.
3. Have the rat handler place two rats in a transfer cage.
4. Place the rat in the running wheel.
5. Have the group member with the stopwatch begin the time.
6. Allow rat to run on running wheel for 10 minutes.
7. After 10 minutes is over, record the number of laps taken.
8. Remove rat from running wheel and place back in transfer cage (allow 2-3 minutes
between time trials to transfer rats to and from the running wheel)
9. Repeat steps 3-8 for each rat (10 min per rat X 8 rats = 80 min).
10.
Total time per experiment day: 10 min + 10 min + 24 min + 80 min = 124 min
Note: The instructions above are for the first three lab periods. The protocol for weeks 4-5 are
on the subsequent page.

2. Protocol for weeks 4-5: Cognition With Morris Water Maze
Total experiment time given:160 min.
Materials:
● water tank filled with water and adjusted to room temperature
● water maze platform, non-toxic white paint
● 4 clean towels
● transfer cages (4) with lids
● paper towels to line transfer cages
● stopwatch
● clipboard
● pencils or pens
● observation sheets
● net and container for pooper-scooping
● broom for stirring
● laptop with Any-Maze software
● pool thermometer
Set up (Will take 15 min on day of experiment):
● Place water maze platform in the tank at the desired position, and fill tank to one inch
above the platform the day before experimentation begins. Filling the tank will take 30-45
min.
● Check water temperature right before experimentation and adjust as necessary - should
be 24 degrees C. Remove thermometer from tank.
● Add approximately ¾ bottle of non-toxic white paint to the tank and stir with the broom to
distribute evenly.
● Line 4 transfer cages with paper towels.
● Set up and turn on the laptop, open Any-Maze, and load the experiment file.
● Place observation sheets in clipboard and have pens or pencils ready.
Instructions:
1. Assign one group member to be the rat handler, another to manage the computer, and a
third to take notes on paper and use the stopwatch to time the trials. Other group
members may help with keeping the rats organized, cleaning, supplies, and pooper-
scooping.
2. Place rats in the transfer cages in the order in which they will complete their water maze
trials. Allow rats to habituate for 10 min.
3. Place rat on platform in the water tank for 30 seconds.
4. Place the rat in the water at the starting position indicated by the observation sheet. The
rat should be place in the water facing the wall of the tank. Let go of the rat and move
away from the side of the tank immediately. The group member managing the computer
should start the tracker while the rat is still at the starting position but after the rat

3. handler has left the field of view of the camer. The group member with the observation
sheet and stopwatch should independently time how long it takes the rat to find the
platform.
5. Allow the rat 60 seconds to swim in the tank and try to find the platform. If the rat cant
find the platform in 60 seconds, the rat handler should guide him to the swim to the
platform and help it climb onto the platform.
6. Record how long it took the rat to find the platform. Record 60 seconds if the rat did not
find the platform on its own.
7. Leave the rat on the platform for 15 seconds before removing him onto a towel with a rat
handler, and then placed into a transfer cage to dry off.
8. Repeat steps 3-7 for each rat (2 min per rat X 8 rats = 16 min) This completes 1 trial
9. Repeat steps 4-7 for each rat to complete a total of 6 trials (2 min per rat X 8 rats X 5
more trials = 80 min).
NOTE: Allow 1-2 min between rats in the maze for grabbing, recording or data, and setup of rat.
(Approximately 48 min total).
Total time per experiment day: 15 min + 80 min + 48 min = 143 min