Validation of an Effective Protocol for Latreille (Diptera: Ceratopogonidae) Detection Using eDNA Metabarcoding.

eDNA metabarcoding is an effective molecular-based identification method for the biosurveillance of flighted insects. An eDNA surveillance approach maintains specimens for secondary morphological identification useful for regulatory applications. This study identified species using eDNA metabarcoding and compared these results to morphological identifications of trapped specimens. Insects were collected using ultraviolet (UV) lighted fan traps containing a saturated salt (NaCl) solution from two locations in Guelph, Ontario, Canada. There were forty-two specimens collected in total. Molecular identification detected four species, , , and Using morphological identification, two out of these four taxonomic ranks were confirmed at the species level ( and ) and one was confirmed at the subgenus level ( []). No molecular detection of species occurred in traps with an abundance of less than three individuals per taxon. The inconsistency in identifying specimens to the species level punctuates the need for curated DNA reference libraries for . In conclusion, the saturated salt (NaCl) solution preserved the morphological characteristics and the eDNA.