What we can find in what's left behind: DNA metabarcoding of semi-aquatic insect exuviae

Semi-aquatic insects contribute to critical ecological functioning of freshwater habitats in both the aquatic and terrestrial phase of their life. Traditional methods of studying their emergence rely on the capture of insects as they emerge, and morphological identification with taxonomic keys. This is not only time consuming but can have large impacts on the study population, obstacles that can be removed by the use of DNA. This study investigated the potential of using exuviae collected from the water surface as DNA source. Both emergence trap samples and insect exuviae were collected from a constructed wetland and a small creek in southern Ontario. Metabarcoding provided a total of 40 samples with 254 distinct taxa from emergence traps (26 samples), and 135 from exuviae (14 samples). There were many similarities between both two sample types, especially for the rich chironomid diversity. Nonetheless, the exuviae samples were able to identify more orders containing semi-aquatic insects. Furthermore, they showed a higher level of diversity within these orders. This higher level of diversity seen in exuviae samples may be due to limitations of emergence traps, such as they only account for a small defined surface area. In contrast, exuviae are representing a much larger area and are free floating, thus collection of emerging taxa is not limited to the emerging site. We were able to show that identification of emerging aquatic insects through metabarcoding of exuviae is a useful method for the study of insect emergence.