 The study aimed to develop universal PCR primers for meta-barcoding environmental DNA from fishes. Primers were designed using aligned whole mitochondrial genome sequences from 880 species, supplemented by partial mitogenome sequences from 160 elasmabrunx, sharks and rays. The primers target a hypervariable region of the 12 SRNA gene, 163-185 BP, which contains sufficient information to identify fishes to taxonomic family, genus and species except for some closely related congeners. To test the versatility of the primers across a diverse range of fishes, Edna was sampled from four tanks in the Okinawa Churomi Aquarium with known species compositions, prepared dual-indexed libraries and performed paired-end sequencing of the region using high-throughput next-generation sequencing technologies. Out of the 180 marine fish species contained in the four tanks with reference sequences in a custom database, 168 species, 93.3%, were detected distributed across 59 families and 123 genera. The meta-barcoding approach presented here is non-invasive, more efficient, more cost-effective and more sensitive than the traditional survey methods. It has the potential to serve as an alternative, or complementary, tool for biodiversity monitoring that revolutionizes natural resource management and ecological studies of fish communities on larger spatial and temporal scales. This article was offered by MMEA, WISOTO, T. Fukunaga, and others. We are article.tv, links in the description below.