Implementation of the 3Rs (Replacement, Refinement and Reduction) in animal studies is a legal requirement in many countries. In The Netherlands, animal welfare officers (AWOs) are appointed to monitor the welfare of laboratory animals. As part of this task, AWOs give advice to researchers and can therefore have an influential role in implementing 3R methods in research. A national survey was conducted to gain more insight into how Dutch AWOs obtain and apply 3R information in their daily work. Nearly half of the AWO population filled out the questionnaire (15/32; a response rate of 46.9%). Two-thirds of the respondents pointed out that finding 3R information is not an easy task and more than half of the respondents believed that information on possibilities to implement the 3Rs is regularly being missed. The respondents indicated that most 3R information is obtained directly from colleagues and other AWOs. Special online 3R databases are rarely used. All the responding AWOs feel that they contribute to Refinement (15/15), nearly one-third of the respondents feel they contribute to Reduction (4/15), and one AWO feels he/she contributes to Replacement (1/15). According to the respondents, better exchange of knowledge can contribute to more successful implementation of the 3Rs. How this knowledge exchange can best be established and facilitated needs further exploration. To this end, the authors make suggestions for a 3R-integrated evidence-based approach.