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The Y-shaped maze consists of a transparent acrylic tank with dimensions designed as follows: the starting arm measures 18 cm in length and 10 cm in width, while each of the goal arms extends to 30 cm in length and 5 cm in width. The entire structure stands at a height of 20 cm. All three arms are fitted with 1 cm diameter polyethylene tubing at their intake (goal arms) and outflow (start arm) openings. To regulate and measure fluid flow rates, a flow meter is positioned at the intake opening. The inflow of fluid is achieved through the use of elevated water cylinders (utilizing gravity), which are connected to the tubing system.
The experimental subjects should be housed in tanks that are consistently maintained at a stable temperature and pH level. These environmental conditions should be rigorously upheld throughout the experimental sessions to ensure the well-being and minimize stress for the fish. Adequate aeration of the water and maintaining its quality at optimal levels are equally crucial.
The Y-Maze experimental tank should be filled with water up to a depth of 7 cm to ensure minimal stress on the fish during trials. Position the Y-Maze tank inside a 50 cm tall opaque plastic enclosure placed on Styrofoam sheets to eliminate external visual cues and vibrations. Illuminate the tank from above for clear observation. For automated tracking and video recording of fish behaviors and movements, utilize specialized software like Noldus Ethovision XT installed on a ceiling-mounted system above the tank.
Between each experimental subject, ensure thorough emptying and rinsing of the tank to maintain cleanliness and eliminate residual influences.
The subjects are to undergo individual exposure to the maze for a duration of 10 minutes each day over a period of 3 consecutive days. To commence the behavioral testing, position the subject behind a mesh sliding door located at the start arm of the maze. The subject should be released into the maze within a 30-second timeframe, coinciding with the initiation of water flow. Testing should conclude after a duration of 5 minutes, following which the subject is promptly returned to its original tank habitat.
In assessing preferences and avoidance using the Y-Maze, subjects underwent testing involving both positive and aversive stimuli across separate experiments. The positive stimulus consisted of a tank containing conspecific shoals, matched in size and weight to the subjects, positioned adjacent to one goal arm wall. Denatonium benzoate served as the aversive stimulus, administered at a concentration of 2.5nM with a maintained flow rate of 0.4 cm/sec.
Subjects were divided into experimental and control groups for both conditions. Results indicated that subjects spent significantly more time in the arm adjacent to the shoal-containing tank compared to when no shoals were present. Conversely, in the aversive stimulus experiment, subjects displayed a preference for the freshwater arm over the arm containing denatonium benzoate, spending more time than expected by chance (Grella et al., 2010).
The following data can be observed in the flow-through Y-Maze tasks,
Grella, S. L, Kapur, N., & Gerlai, R. (2010). A Y-maze Choice Task Fails to Detect Alcohol Avoidance or Alcohol Preference in Zebrafish. International Journal of Comparative Psychology, 23(1)
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