Cetacean watching is an increasingly popular economic activity in Ecuador for tourism operators. Despite government regulations, cetacean watching can have a negative impact on observed animal populations. To enhance good dolphin-watching practices, a course was carried out in Puerto El Morro, Ecuador about sensitisation activities promoting empathy towards and knowledge about bottlenose dolphins, local avifauna and mangrove ecosystems. The course provided tourism operator staff with theoretical and practical knowledge on dolphin physiology and ecology, with a focus on empathy towards the species and on regulations to be respected during the watching tours. The course included interactive workshops, didactic materials, advice, video screening and structured conversations. Two innovative questionnaires were implemented on the boat during 68 dolphin-watching tours to evaluate whether the training course had improved the regulation compliance and on-tour guiding quality of the tour guides and boat drivers. The results showed a statistically significant improvement in dolphin-watching practices in compliance with the “Regulations for the Whale and Dolphin Watching of Ecuador” (p = .0002) and in guidance quality (p = .0004) after the training course. Boat drivers were identified as influential actors in compliance with regulations and should also be included in the environmental awareness training courses. The study showed that empathy-based sensitisation and knowledge reinforcement positively affect policy compliance and can generate new sustainable approaches for future dolphin-watching activities.