Sylvie Cloutier, Kim Wahl, Chelsea Baker, and Ruth C. Newberry

ABSTRACT: Handling small animals for veterinary and experimental procedures can negatively affect animal wellbeing. We hypothesized that playful handling (tickling) would decrease stress associated with repeated injections in adult laboratory rats, specially those with prior tickling experience. We compared responses of 4 groups of male Sprague–Dawley rats to intraperitoneal injection of saline daily for 10 d. Rats either tickled or not tickled as juveniles (2 min/d for 21 d) were exposed as adults to either a passive hand or tickling for 2 min immediately before and after injections. Rates of vocalization (22- and 50-kHz ultrasonic vocalizations (USV), indicative of negative and positive affective states, respectively, and audible calls indicative of pain and discomfort) were quantified before, during, and after injection. Tickling before and after injection, especially when combined with juvenile tickling experience (ending 40 to 50 d earlier), increased 50-kHz USV rates before and after injection, reduced audible call rate during injection, and decreased the duration of the injection procedure. The treatments did not affect indicators of physiologic stress (body weight change; fecal corticosteroid levels). We conclude that playful handling performed in association with a mildly aversive procedure serves as a useful refinement by inducing a positive affective state that mitigates the aversiveness of the procedure and makes rats easier to handle, specially when they have been accustomed to tickling as juveniles.