DNA Barcoding Reveals a Distinct Mediterranean Mitochondrial Lineage of Lasiocampa tripolitania from Libya Based on COI Sequences: Implications for Taxonomy, Conservation, and Forensic Identification

Abstract

DNA barcoding is widely used for organism identification because the mitochondrial cytochrome c oxidase subunit I gene, COI, usually separates closely related species and geographic lineages. In this study, we examined the COI barcode position of the Libyan moth Lasiocampa tripolitania using the GenBank sequence AM397633.1, reported from near Zuwara, Libya. The final aligned dataset included a 576 bp COI fragment, three closely related Mediterranean Lasiocampa sequences from Malta, 32 European sequences assigned to Lasiocampa quercus and Lasiocampa trifolii. Phylogenetic trees were reconstructed in MEGA 10 using Neighbor-Joining and Maximum Likelihood under the Tamura 3-parameter model with 1000 bootstrap replicates. Pairwise Tamura 3-parameter distances and ASAP delimitation were also used. The Libyan sequence clustered within a Mediterranean Lasiocampa lineage and was most closely related to Maltese Lasiocampa sequences, but it formed a distinct Libyan branch. Mean distance between Mediterranean Lasiocampa and European L. quercus was 0.163, while the distance from European L. trifolii was 0.119. These results support a distinct Mediterranean mitochondrial lineage, but broader sampling is required before formally finalizing the taxonomic revision