Standard Operating Procedure for Spray Test
Grooming behavior is assessed with a spray test between 12pm and 1pm on observation days. Mice are removed from their cages and sprayed with a single mist of water. They are placed in clean cages with ad-lib food and water available. Their behavior is recorded for 15 minutes following the spray in the cage using digital video system. Behavior from the 15 minutes following the spray is assessed in the video recordings by trained viewers. Focal sampling and continuous recording methods are used. Grooming behaviors are focused in on the ethogram and were adapted from Berridge et. al. 2005 and are described as follows:
Manual Grooming - the animal grooms its muzzle face and head using its front paws, alternating left and right. The animal typically performs this behavior while stationary and seated, but occasionally while rearing. This behavior is often followed by oral grooming.
Oral Grooming - The animal grooms its body by licking usually beginning on the upper back and neck, then extending down to more caudal areas of the body, including the tail.
Scratching - the animals uses its hind-limbs to scratch its head, neck, and back. These scratches are very fast and bouts are usually short-lived. Record the number of scratches and the region that was scratched in addition to the typical behavioral data.
Only grooming behaviors involving 2 or more grooming movements in succession were counted. Non-grooming behaviors were also assessed, including active (all other behavior), inactive (sleeping/rest) and not visible (mouse was performing behavior but the viewer was unable to assess grooming behavior). Pauses lasting less than 3 seconds are not counted.