Amphimerus, a liver fluke, is the causative agent of amphimeriasis, a foodborne disease acquired thought the consumption of infected raw or undercooked river fish—a practice embedded in traditional culinary customs. Amphimeriasis represents a significant public health issue and has been endemic in Ecuador since 2011, particularly among the Chachi Amerindians and Montubios populations residing in tropical ecoregions. By employing a mixed-methods research design, we conducted a community-based, cross-sectional study. A survey comprising of 63 questions on KAP was administrated in person to community members, health personnel, and academic staff in the two populations. Additionally, 67 semi-structured interviews were performed. Microscopy was achieved on 273 human and 80 dog fecal samples to detect Amphimerus eggs. A total of 86 questionnaires (54 Chachi) and 67 interviews (44 Chachi), out of 300 residents, were completed. Among the respondents, 31.4% were aware of Amphimerus, locally referred to as “liver worm”. Although 79.1% reported not consuming raw fish, most admitted eating raw fish with lime juice and salt, a preparation known as “curtido”, and 59.3% reported consuming smoked fish. Here, 86.1% of participants considered “liver worm” a serious disease, and 55.8% recognized raw or marinated fish as a potential transmission route. The Chachi showed a preference for smoked fish, whereas the Montubios favoured “curtido”. The prevalence of Amphimerus infection was 23% in humans and 16.2% in dogs. Differences in KAP were observed between infected and non-infected individuals. Local health and academic personnel demonstrated insufficient knowledge about amphimeriasis. Some religious individuals refrained from participating, stating that they were “with God”. Despite the high prevalence of Amphimerus infection in both humans and dogs, knowledge about the parasite, the disease, and its transmission routes remains limited. Health education initiatives should be designed to modify the population’s KAP. It is crucial for national and local health authorities, as well as religious leaders, to be informed and actively involved in the prevention and control of amphimeriasis.
