HIGH-RESOLUTION MELTING AS A MOLECULAR TOOL FOR CETACEAN SPECIES IDENTIFICATION IN THE MEDITERRANEAN SEA

ANALISI DI MELT AD ALTA RISOLUZIONE COME STRUMENTO MOLECOLARE PER L'IDENTIFICAZIONE DELLE SPECIE DI CETACEI NEL MAR MEDITERRANEO

Abstract

Effective species identification underpins cetacean conservation in the Mediterranean Sea, where several populations are threatened. We developed a High-Resolution Melting (HRM) protocol to rapidly and cost-effectively identify four representative cetacean species: the bottlenose dolphin (Tursiops truncatus), the striped dolphin (Stenella coeruleoalba), the sperm whale (Physeter macrocephalus), and the fin whale (Balaenoptera physalus). Species-specific primers targeting mitochondrial cytochrome b and D-loop regions produced distinct melting profiles. Validation on tissue and fecal samples confirmed high sensitivity, reproducibility, and discrimination, with an accuracy of over 95% in identifying unknown samples. Compared to sequencing, HRM is faster, cheaper, portable, and effective on degraded material, including samples from stranded individuals. This method represents a valuable tool for non-invasive genetic surveys and real-time monitoring, enhancing conservation efforts and enforcement of regulations against illegal trade.