The fishing catPrionailurus viverrinus is a medium-sized wetland specialist felid threatened primarily by habitat loss, human–wildlife conflict, and road mortality. Limited information is available on their life history in human-modified environments. We investigated the dietary habits of fishing cats within a human-dominated landscape in southern Nepal to inform conservation strategies that promote co-existence with local communities. We carried out a comprehensive analysis of fishing cat scats through microscopic analysis and DNA metabarcoding. We found fishing cats to have a diverse diet including fishes, mammals, birds, reptiles, amphibians, mollusks, and insects. The trophic niche breadth of 0.68 suggests a generalist feeding behavior (> 0.5). Remains of domestic or commercially farmed animals (including fish) were present in approximately one-half of the scat samples. A large contribution of small mammals and birds to the fishing cat diet further indicates their role in regulating rodents and other pests in agricultural areas. We suggest that highlighting these findings through community education and awareness can change farmers’ negative perceptions, reducing threats of retaliatory killing and increasing support for conservation efforts.