2.3.2. Elevated Plus Maze (EPM)

The EPM was applied to detect anxiety [24,25]. The device consists of a 51.3 cm-high wooden maze. The dimensions of the open arms were 30 cm of length to 5 cm of width, and a small 0.5 cm-high edge. The dimensions of the closed arms were 30 cm of length to 5 cm of width, and a 15 cm-high wall. At the beginning, the mouse was placed in the middle field facing an open arm, and the middle field was defined as a separate area. The test was recorded for five minutes. The first analyses for all groups started with the following parameters: number of total entries, number of closed-arm entries, percent of open-arm entries (vs. total number of entries), and percent of open-arm time (vs. total tested time). To enhance the level of validity, a second analysis was performed, including the following ethological parameters: rearing, percent of protected head dip, amount of total head dip, and number of piles of feces. Protected head dip is a risk assessment behavior. The mouse leans from the center or closed-arm area forward to the center or open-arm area. It can be used to detect an anxiogenic effect even in the absence of changes in conventional open-arm measurement [24,25,26].