High‐Throughput <scp>DNA</scp> Sequencing Reveals Gastric Content Composition and Inter‐Specific Variation in <i>Pampus</i> Fishes

<i>Pampus</i> species play a significant role in offshore food webs and are regarded as representative economically important fish. Owing to variations in their geographical distribution, distinct differences in feeding habits exist among pomfret species. However, the presence of an esophageal sac in pomfrets complicates accurate identification of their dietary composition through conventional stomach content analysis. Consequently, research on the feeding ecology of pomfrets remains limited and imprecise. This study employed environmental DNA (eDNA) high-throughput sequencing technology to analyze the composition and relative abundance of gastric contents in three <i>Pampus</i> species (<i>P. argenteus</i>, <i>P. punctatissimus</i>, and <i>P. cinereus</i>) collected from Qingdao, Shandong Province and Wenchang, Hainan Province. Key findings include: (1) Analysis revealed 21, 7, and 2 fish species in the gastric contents of <i>P. argenteus</i>, <i>P. punctatissimus</i>, and <i>P. cinereus</i> respectively, with anchovy (<i>Engraulis japonicus</i>) demonstrating the highest relative abundance in the former two species, while <i>Thryssa hamiltonii</i> predominated in <i>P. cinereus</i>. (2) Crustacean communities comprised 49, 42, and 26 species across the respective species, constituting over 60% of total identified taxa and indicating their trophic importance. <i>Acetes japonicus</i> exhibited maximal relative abundance in <i>P. argenteus</i> and <i>P. punctatissimus</i>, whereas <i>Acetes chinensis</i> dominated in <i>P. cinereus</i>. (3) Cephalopod analysis identified 8 and 6 species in <i>P. argenteus</i> and <i>P. punctatissimus</i> respectively, with <i>Loligo beka</i> and <i>Sepiola birostrata</i> showing peak abundance in each species, while no cephalopod species were detected in <i>P. cinereus</i>. Comparative analysis with traditional dissection methods and stable carbon-nitrogen isotope techniques confirmed crustaceans as the most diverse dietary component across <i>Pampus</i> species. This investigation demonstrates the efficacy of eDNA metabarcoding for elucidating feeding ecology in <i>Pampus</i> spp., providing critical insights into their trophic interactions and ecological significance within marine food webs.