Environmental DNA (eDNA) metabarcoding emerges as a powerful method, allowing a more exhaustive investigation of fish fauna than any other methods. Yet, the general use of eDNA as a replacement of traditional methods such as visual surveys or physical sampling remains debatable. Therefore, a prior understanding of eDNA’s spatial resolution is necessary. This study aimed to compare the assessments of fish diversity at three spatial scales by eDNA, underwater visual census (UVC), and diver-operated video (DOV) surveys across 21 reef sites in northern Taiwan. The specific objectives were to explore the regional species pool (γ-diversity), reveal spatial patterns of fish assemblages (β-diversity), and disentangle the relationships between fish assemblages and benthic composition (α-diversity). At the γ-diversity level, a total of 438 marine fish species were detected across methods. eDNA exhibits an extraordinary power to explore the regional species pool given sufficient replication, a power which is unachievable by DOV and UVC. At the β-diversity level, all the methods successfully revealed the same spatial patterns of beta diversity and the distance decay of similarity in fish assemblages. At the α-diversity level, none of the methods is capable of investigating the entire resident fish fauna, but visual surveys are more suitable for scrutinizing interactions between fish and benthos. Instead of undiscriminatingly recommending a combination of eDNA with traditional survey methods, we suggest implementing specific surveys in accordance with the ecological questions of interest.
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