Towards routine DNA metabarcoding of macroinvertebrates using bulk samples for freshwater bioassessment: Effects of debris and storage conditions on the recovery of target taxa

• DOI: 10.1111/fwb.13443
• Publication Date: 2019-12-20T05:30:18Z
• Authors: Susan J. Nichols; Ben J. Kefford; Catriona D. Campbell; Jonas Bylemans; Eloise Chandler; Jonathan P. Bray; Michael Shackleton; Katie L. Robinson; Melissa E. Carew; Elise M. Furlan
• Keywords: Environmental DNA

Abstract Macroinvertebrates are commonly sampled for bioassessment of freshwater ecosystems. However, current protocols involve laborious sorting the animals from debris (sample matrix) and morphological identification, where species level identifications often difficult. DNA metabarcoding has potential to improve by reducing time taken process samples accuracy speed macroinvertebrate identification. In this study, we evaluated samples, which include macroinvertebrates collected in sample nets, test if bulk, unsorted can be used assess diversity. First, tested matrix prevented detection six target taxa when metabarcoding. Second, storage influenced same macroinvertebrates. We also explored different levels replication at sample, sub‐sample, polymerase chain reaction compared overall families detected using those identified morphologically. found that presence did not interfere with or inhibit taxa. Furthermore, various methods affect detection. The reliability improved as hierarchical were combined. strong overlap between family diversity comparing Extracting bulk included could removing need samples. was dependent on up 1 year 95% ethanol, room temperature after heating. had advantage identifying species, but good barcode libraries needed widespread identifications. Further investigation should focus including multiple composition densities refine standardise processing protocols, building comprehensive aquatic