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The pilot study in Bhutan's Mangde Chhu river basin demonstrates that eDNA metabarcoding can detect a wide array of vertebrate species, including rare and IUCN-listed threatened species, often complementing or exceeding the detections of traditional camera trapping. While the study identified 134 species and 201 unique vertebrate taxa, it highlighted significant gaps in the NCBI reference databases for birds and reptiles. The findings suggest eDNA is a cost-effective tool for monitoring high-value species like the tiger and Golden mahseer, though the authors recommend establishing national laboratories and expanding DNA databases to fully operationalize a national biodiversity monitoring framework.