Identification of Necrophagous Beetles (Coleoptera) Using Low-Resolution Real-Time PCR in the Buffer Zone of Kampinos National Park

Forensic entomology is concerned with the study of arthropods that assist in criminal investigations. Among them, necrophagous beetles (Coleoptera) play a crucial role as they are found in all stages of decomposition of corpses, which makes them valuable for forensic analysis. Accurate species identification is crucial for the effective use of entomological evidence. Efficient, cost-effective, and rapid methods are essential for this purpose. In the present study, we developed a novel assay that enables simple, inexpensive, and rapid identification of necrophagous beetle species. Using two primer sets targeting the cytochrome c oxidase 1 (COI) gene and analyzing the resulting SYBR Green I melting curves, we were able to identify fourteen beetle species: <i>Aleochara curtula</i> (Goeze, 1777), <i>Anoplotrupes stercorosus</i> (Scriba, 1791), <i>Creophilus maxillosus</i> (Linnaeus, 1758), <i>Dermestes undulatus</i> (Brahm, 1790), <i>Hister unicolor</i> (Linnaeus, 1758), <i>Margarinotus brunneus</i> (Fabricius, 1775), <i>Necrodes littoralis</i> (Linnaeus, 1758), <i>Nicrophorus investigator</i> (Zetterstedt, 1824), <i>Ontholestes murinus</i> (Linnaeus, 1758), <i>Oiceoptoma thoracicum</i> (Linnaeus, 1758), <i>Philonthus cognatus</i> (Stephens, 1832), <i>Saprinus planiusculus</i> (Motschulsky, 1849), <i>Silpha tristis</i> (Illiger, 1798), and <i>Thanatophilus rugosus</i> (Linnaeus, 1758). Data on the melting temperature of the amplicon can be obtained within 60-90 min after DNA extraction. This study expands the understanding of the species composition of necrophagous Coleoptera, which is crucial for forensic entomology methods based on insect succession. Furthermore, it provides the first comprehensive data on necrophagous beetles in the vicinity of Kampinos National Park.