This study examines the evidence regarding the effectiveness of interventions aimed at the prevention of harmful alcohol and drug use in nightlife settings. A literature search was conducted in 2009. Of 161 studies, 17 experimental studies were included in the review, 15 alcohol-related and two drug-related studies. The results show that preventive substance abuse interventions in nightlife settings can effectively reduce high-risk alcohol consumption, alcohol-related injury, violent crimes, access to alcohol by underage youth, and alcohol service to intoxicated people. A combination of approaches such as enforcement activities seem to be facilitating factors. However, results should be considered cautiously as more gold standard (cost-) effectiveness research is required, in particular directed at drug prevention and educational interventions in nightlife settings.