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Abstract As many ecosystems are under increasing pressure from invasive species, habitat degradation, overgrazing and overharvesting, pollution, and climate change, dietary niche monitoring is gaining importance. The Malai Mahadeshwara Wildlife Sanctuary (MMH) in southern India is home to several long‐standing ethnic and tribal groups and supports a considerable number of domestic herbivores (cattle, goats and water buffalo) as well as a range of wildlife (including several species of deer, bonnet macaque, and Asian elephant). We reconstructed dietary niche partitioning of the herbivores occurring in MMH using eDNA metabarcoding to quantify diet richness, composition, and overlap. In total, we distinguish 134 diet items (molecular operational taxonomic units), covering 31 plant families. Overall, our results indicate 35% overlap in domestic and wild herbivore diet items. The greatest overlap is found for the dietary niches of cattle and sambar deer (Pianka's niche overlap index: 0.68), and the dietary niche of cattle also overlaps considerably with those of Indian hare (0.65) and Asian elephant (0.46). This suggests that these herbivores may compete for these food plants in the case of limited availability, which could lead to exclusion of some herbivore species. Particular concern should go to bonnet macaque and Asian elephant as their below average dietary richness could make them vulnerable to changes in their environment. With increasing pressures on local wildlife from a range of different factors, DNA metabarcoding of fecal samples is a non‐invasive method for monitoring changes in animal diets, providing valuable information for the management of biodiversity in mosaic natural and anthropogenic landscapes.