Effects of Clicker Training on Behavioral and Stress Markers of Welfare in the F1 Generation of CD1 Mice: A Pilot Study

Training in laboratory animals is important to ensure that scientific studies are reliable, reproducible, and ethically acceptable. Well-trained animals experience less stress and exhibit fewer unwanted behaviors, improving both welfare and research outcomes. Clicker training is widely used in animal training as a positive reinforcement method to reduce distress. The present study was designed as a pilot study to examine the effects of a four-day clicker training protocol applied to both dams and their offspring, with behavioral outcomes assessed exclusively in the offspring. The results indicated that clicker training potentially increased voluntary interaction with the experimenter and promoted body weight gain during the training period. No significant effects of offspring training were found for classical anxiety-related measures (EPM open arm time, OF center time, Nest Building Test, Sucrose Preference Test, Forced Swim Test) or plasma corticosterone. A potential sex effect was observed across locomotor, center-zone, and corticosterone measures. Maternal training effects on offspring outcomes should be interpreted as preliminary exploratory observations, as only two dams per maternal group were available. Given the exploratory nature and limited sample size of this pilot study, the findings should be interpreted with caution. Under the present condition, clicker training produced a context-specific improvement in human–animal interaction without evidence of harm, but did not produce generalized reductions in anxiety-like behaviors as assessed by standard paradigms. Further studies with larger sample sizes are needed to confirm these findings.