COI Insights into Diversity and Species Delimitation of Immature Stages of Non-Biting Midges (Diptera: Chironomidae).

The diversity of non-biting midges (Chironomidae, Diptera) remains an unresolved topic, with estimates of species numbers ranging from 6000 to 15,000 according to various authors. To assess Chironomidae diversity in Lithuania, we evaluate the effectiveness of COI gene-based species delimitation methods for providing rapid diversity estimates. Nevertheless, differences between tree-based and distance-based approaches can result in varying group classifications, which may cause species numbers to be overestimated or underestimated. For our study, we analyzed a dataset of 109 specimens sampled from six Lithuanian streams. By applying multiple methods, such as Assemble Species by Automatic Partitioning (ASAP), Automatic Barcode Gap Discovery (ABGD), the generalized mixed Yule-coalescent (GMYC) model, and the Bayesian implementation of the Poisson Tree Processes (bPTP) model, we found that species estimates ranged from 28 to 58. Among these methods, ASAP proved to be the most effective for our dataset, identifying 58 putative species. These results reinforce our assumption that the current understanding of Chironomidae species diversity is incomplete.