MONITORING THE DISTRIBUTION OF APIS MELLIFERA MITOCHONDRIAL DNA LINEAGES OVER EIGHT YEARS IN THE EMILIA-ROMAGNA REGION USING HONEY EDNA

Honey contains DNA derived from all organisms involved in its production, including honey bees (Apis mellifera), making it a non-invasive and valuable source of genetic information for honey bee population genetic analysis. In this study, we used DNA assays we previously developed to identify the mitochondrial DNA lineages of the honey bees that produced honey. These assays differentiate three mitochondrial DNA (mtDNA) lineages (A, C, and M) of Apis mellifera and distinguish two mitotypes within the C lineage: C1 and C2, predominantly associated with A. m. ligustica and A. m. carnica, respectively. This study included a total of approximately 1200 honey samples produced from 2018 to 2025 in the Emilia-Romagna region (in the North of Italy). The C1 mitotype was the most frequent, indicating the ongoing prevalence of A. m. ligustica in regional beekeeping. Some samples carried the C2 mitotype, and a few contained both C1 and C2, suggesting genetic heterogeneity. Additionally, a small number of honey samples included A and/or M lineages in addition to the C lineage. Geographical data from the honey samples allowed us to map mitotype distribution across the Emilia-Romagna region and evaluate the overtime modifications in local honey bee populations. These findings offer valuable information for developing conservation strategies for honey bee genetic resources and preserve the genetic integrity of local A. mellifera populations.