Effect of Repeated Exposure to Isoflurane on Nest Building and Burrowing in Mice

Nest building and burrowing are highly motivated natural behaviors in rodents, and changes in these behaviors can serve as welfare assessment tools. In this study, we investigated: 1) the limits of agreement between 2 observers for a refined scoring method for nest-building behavior; 2) the effect of repeated exposure to 15 min of isoflurane on nest-building behavior; 3) the effect of 24 h of grid-floor housing, repeated exposure to 15 min isoflurane, and daily intraperitoneal injection of 0.2 mL 0.9% isotonic saline for 3 d on burrowing behavior; and 4) the effect of exposure to grid-floor housing, isoflurane, and intraperitoneal injections on fecal corticosterone metabolites, body weight, fur status, and sucrose preference in mice. SPF C57BL/6NTac female mice (n = 27) were included in the study and were assessed first for burrowing behavior, followed by 2 wk of rest and then for nesting behavior. The refined scoring method for nest-building activity had good inter observer agreement. According to this method, a single exposure to anesthesia with isoflurane led to a decrease in nest-building activity and sucrose preference; a second exposure to anesthesia with isoflurane had no effect on nest building. Neither grid-floor housing nor repeated exposure to isoflurane anesthesia had any effect on burrowing behavior in mice. In contrast, intraperitoneal injections increased burrowing behavior. In conclusion, a refined scoring method for nest-building activity test that we developed for this study proved to be objective and sensitive to the effect of an initial exposure to anesthesia with isoflurane.