![]() This protocol was followed by two common research procedures: subcutaneous injection and tail vein blood sampling. The training protocol was designed to habituate the mice to the procedures involved in subcutaneous injection, e.g., removal from the cage, skin pinch. Here, two groups of CD1 mice were exposed to either traditional laboratory handling methods or a training protocol with cup lifting for 3 and 5 weeks. Furthermore, simple methods to evaluate distress in mice are lacking, and specialized behavioral or biochemical tests are often required. Mice are the most commonly used laboratory animal, yet there are limited studies which investigate the effects of repeated handling on their welfare and scientific outcomes. Based on our findings, we hypothesize that clicker training is an effective tool to motivate mice and increase performance on the CatWalk test without potentially impairing their welfare (e.g., by puffing them). ![]() Clicker training lowered anxiety-like behaviors in our mice, especially in females, where a significant difference was observed between trained and untrained ones. In addition, clicker training reduced the number of runs required by mice, which was more pronounced in males. In the CatWalk test, we observed that clicker training improved the running speed of the mice. In this study, we trained male and female mice to complete the CatWalk protocol while assessing the effects of training on their well-being (Open Filed and Elevated Plus Maze). We have already demonstrated the positive effects of clicker training. Positive reinforcement training of laboratory animals is a thriving tool for refinement and contributes to meeting the demands instituted by Directive 2010/63/EU. ![]() ![]() However, such stimuli are usually aversive and can impair welfare. Mice usually require a stimulus to walk to the end of the catwalk. The CatWalk test relies on the run of mice across the platform to measure a constant speed with low variation. By developing a reliable protocol that can be easily integrated into the daily routine of the keeping of laboratory mice, the lifetime experience of welfare in the mice can be improved substantially. This study has identified that clicker training for mice strongly correlates with reduced fear in the mice during human-mice interactions, as shown by reduced anxiety-related behaviors (e.g., defecation, vocalization, and urination) and fewer depression-like behaviors (e.g., floating). The mice learned rather quickly when challenged with tasks of the clicker training protocol, and almost all trained mice overcame the challenges they were given (100% of female mice and 83% of male mice). The effective implementation of the clicker training protocol with a cohort of 12 BALB/c inbred mice of each sex proved to be uncomplicated. Clicker training is a form of positive reinforcement training using a conditioned secondary reinforcer, the "click" sound of a clicker, which serves as a time bridge between the strengthened behavior and an upcoming reward. By introducing clicker training to the keeping of mice, not only do we promote the amicable treatment of mice, but we also enable them to experience cognitive enrichment. Previous research determined a profound impact of gentle handling protocols on the well-being of laboratory mice. ![]() Establishing new refinement strategies in laboratory animal science is a central goal in fulfilling the requirements of Directive 2010/63/EU. ![]()
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