Abstract

Background:
Wax moths represent major pests in honeybee colonies, damaging comb structures and reducing colony strength. Morphological and molecular analyses enable precise species identification and clarify their diversity and distribution in Kurdistan.

Aim:
This study aimed to morphologically and molecularly identify wax moth species collected from different locations in Kurdistan, Iraq.

Methods:
Comparison of larval weight, adult body weight, and wing length of Galleria mellonella and Achroia grisella across different locations in the Kurdistan region. Significant geographical variations were observed, indicating that environmental conditions strongly influence the morphological traits of both wax moth species and also molecularly identify wax moth species collected from different locations in Kurdistan, Iraq, using mitochondrial cytochrome coxidase subunit I (COI) gene analysis. Polymerase chain reaction (PCR) amplification using species-specific primers successfully produced ~550 bp fragments from all insect samples, which were confirmed by agarose gel electrophoresis. Partial COI sequences from 12 specimens were generated and edited using the Bio Edit software, and then compared with reference sequences in the National Center for Biotechnology Information using BLAST.

Results:
Morphological traits of wax moths varied significantly across Kurdistan, with Halabja populations showing greater body weight and size, while Penjwen consistently exhibited lower measurements, also the results showed a sequence identity of 99.5%–100% with G. melonella and A. grisella, confirming accurate species identification. All validated sequences were submitted to GenBank and assigned accession numbers. Phylogenetic analysis using the maximum likelihood method in MEGA 12 grouped samples into two distinct clades corresponding to the two species, demonstrating close genetic similarity within each genus. These findings confirm the effectiveness of COI-based DNA barcoding for reliable identification of wax moth species and provide baseline genetic data for future studies on population structure and pest management in the region.

Conclusion:
Significant morphological variation occurred across Kurdistan, with larger wax moths in Halabja and smaller in Penjwen, while COI barcoding confirmed species identity, enabling robust future population studies.

Key words: Achroia grisella; COI region; Galleria mellonella; Wax moth.