Characterizing the spatial and temporal occurrence patterns of the endangered botiid loach Parabotia curtus by environmental DNA analysis using a newly developed species-specific primer set

We report a novel species-specific primer set for detection of the endangered botiid loach Parabotia curtus in environmental waters. A primer test using DNA extracted from tissues of P. curtus and four other loach species indicated its high specificity to P. curtus . Application of the primer set to field samples collected throughout the year in a natural habitat confirmed its ability to characterize spatial and temporal occurrence patterns of this rare, small fish that often inhabits turbid waters.