A Preliminary Study on Species Identification of Immature Necrophagous Phorid Flies Based on FTIR Spectroscopy

Phorid flies serve as the main colonizers of human remains in both indoor and burial environments. Their developmental patterns can be utilized to estimate the minimum postmortem interval (minPMI). Accurate species identification, particularly for immature stages, is essential before utilizing their developmental data to estimate minPMI. This study employed Fourier transform infrared spectroscopy (FTIR) coupled with principal components analysis (PCA) and partial least squares-discriminant analysis (PLS-DA) to investigate species identification of eggs (0 h, 8 h, 16 h), larvae (12 h, 60 h, 84 h), and pupae (1 d, 5 d, 10 d) of three necrophagous Phoridae species, <i>Dohrniphora cornuta</i>, <i>Diplonevra funebris</i>, and <i>Megaselia scalaris</i> at 24 °C. The results showed that the FTIR spectra within the fingerprint region (1800-900 cm^-1) differed among the three immature phorid flies. These differences were primarily manifested in absorption peak intensities. The PLS-DA analysis successfully distinguished the three species at the same developmental stage. This study demonstrated the feasibility of utilizing FTIR spectroscopy coupled with chemometric methods to both rapidly identify the species of immature small flies and simultaneously estimate their age.