Contrary to the limited research on the effect of the testing history on performance, there is a significantly larger number of studies focusing on the effect of test-retest on the reliability of anxiety tests. Exposing 5 strains of mouse to repeated testing on three consecutive days in an elevated zero-maze, showed that anxiety increased over the three trials.[8] An interesting finding of this study was that only a repeated-trial zero maze managed to detect anxiety in strains with visual impairment. Since some of the strains use visual or sensory cues to avoid the open arms of the zero-maze; repeated testing in this apparatus is needed in strains with visual impairment.

Recent studies have focused on another issue concerning the batteries of behavioral tests and this is the intra- and inter-specific correlations between the various tasks.[9] Using the multivariate approach, scientists can analyze the vast volume of behavioral data and draw conclusions regarding any effects between the different behavioral tests. It has been shown that correlations exist between the performance in various independent tests, therefore, the behavioral pattern in one behavioral measure can be predictive of the pattern observed in the following ones.

In what order should I perform my behavioral tests?

The golden rule when it comes to the battery test order is to start with the least stressful test and leave the most stressful of all for last. Tests for anxiety-like and exploratory behavior should always be performed first since they can be significantly affected by any previous experience. On the other hand, any task focusing on the cognitive abilities of the animal should be carried out at later stages. This is crucial since the animal’s cognitive performance can be seriously affected if the animal is not properly adapted to the handling procedure and the experimenter. Most of the published behavioral studies abide by this rule of using the least invasive tests before anything else and leaving the more invasive ones for last.[3][6][10][11] Another very important consideration is to allow multiple days of resting time between tests to decrease carryover effects from prior tests.