Open Field Test
The open-field test was used to analyze the locomotor activity, pain, and anxiety-like behaviors, such as grooming (s), rearing (s), sniffing (s), time into the peripheral zone, and crossings (number of times the line of a square is crossed with all 4 legs)[4]. The apparatus is made of light-grey polyvinyl chloride (PVC) (50 x 50 x 25 cm). This test was performed for 30 minutes in the 1, 7, and 14 days after stress induction and in the first h after the treatment on day 7 post-stress. Illumination (about 40 lux) was provided by a light bulb hanging 60 cm above the apparatus. The temperature of the room was maintained at 22 °C and was a recorded test using AnyMaze® 7.0 software. Between each trial, the apparatus was cleaned with a 30% ethanol solution to avoid odor cues.