OBJECTIVES: To assess use and effect of hand antiseptics in veterinary clinical practice. MATERIALS AND METHODS: Veterinary practice nurses were questioned concerning their use of hand antiseptics, in particular waterless hand rubs. Subsequent clinical trials assessed the effect of single applications of alcohol-based and quaternary ammonium compound-based hand rubs at reducing bacterial counts on the hands of theatre nurses in a neutering clinic. RESULTS: The majority of responding practices used waterless hand rubs (alcohol-based, 67·5% and quaternary ammonium compound-based, 9·5%) as their primary hand hygiene agent and believed them to be effective. 23% of practices favoured an antiseptic hand-wash. In clinical trials, alcohol-based rubs were potentially more effective at reducing bacterial counts than quaternary ammonium compound-based rubs especially in the period immediately after application. However, over 3 hours there was no significant change between these and a control group. There were more adverse skin effects in the group using alcohol-based than in the quaternary ammonium compound-based and control groups. The bacterial counts after application were unaffected by variables such as the number of animals contacted. CLINICAL SIGNIFICANCE: The studies provide useful baseline data for evaluation of efficacy of more frequent applications of the most common antiseptic hand rubs used in veterinary practice.