“…With the ongoing employment of integrative approaches using multiple species delimitation methods, it became clear that neither of these always perfectly matches the morphospecies entities (e.g., Ahrens et al, 2013Ahrens et al, , 2016Dalstein et al 2019;Eberle et al, 2019;Lukic et al, 2021) nor do they rarely ever match among each other (Ranasinghe et al, 2022a). Although metabarcoding approaches use mainly distance based clustering algorithms with predefined thresholds for species circumscription (Callahan et al, 2017;Piper et al, 2019), several different pipelines or "cluster parameter values" are in use that may reveal alternative outcome (e.g., Potter et al, 2017;Alberdi et al, 2018;Creedy et al 2019;Bailt et al, 2020) The threshold values used for sequence filtering and the number of reads for the identified MOTUs have an effect on the assessment accuracy of data (Edgar & Flyvbjerg, 2015;Potter et al, 2017;Meyer et al, 2021;Piper et al, 2019Piper et al, , 2022. The choice of the clustering threshold at 3% pairwise distance, as applied by the majority of studies (Elbrecht et al, 2017;Yu et al, 2012;Alberdi et al, 2018), or at 2% for similar taxa (Beentjes et al, 2019;Rossini et al, 2016;Smith et al, 2005) can have a significant impact on taxonomic inferences.…”