Invertebrates are used in environmental, biology, and science education. However, they can elicit disgust, which can be detrimental for motivational and learning outcomes. In addition, practical work including hands-on interaction with living invertebrates could be a viable way to reduce invertebrate disgust and strengthen state intrinsic motivation. Moreover, Big-Five personality may explain the students' reaction to the exposure with live invertebrates. Therefore, the present study tested the effects of an intervention with living invertebrates on disgust and state intrinsic motivation in a sample of 1861 students (age 10-12 years). Moreover, we tested whether an in-school or an out-of-school intervention was more successful in reducing disgust as compared to the control group. While disgust was measured at pre-and post-test, trait motivation was measured at pre-test and state intrinsic motivation was measured at post-test. Results show that while both treatment conditions reduced disgust more than the control condition, the out-of-school condition had a more pronounced disgust reduction than the school condition. Disgust was negatively related to state intrinsic motivation, but state intrinsic motivation was equally high in both treatment conditions (school and university). Extraversion was negatively and neuroticism was positively related to disgust. Big-Five personality was generally associated with state intrinsic motivation. In conclusion, invertebrate disgust can be successfully reduced by an intervention with live invertebrates. The workstation-based intervention with live invertebrates is useful in different settings, both in and out of school and effective with invertebrate species, especially with snails and mealworms, because children's disgust for these species was significantly reduced and lower disgust was related to higher state intrinsic motivation.