An analysis of juvenile hormone bisepoxide III in three species of forensically important flies

Abstract Forensic entomology utilizes the age of the larvae to estimate the minimum post-mortem interval (mPMI), a critical aspect of death where improving the accuracy and reliability of developmental age estimated is essential. Traditional method often rely on larval length, which can be influenced by preservation method, environmental and genetic factors. This study investigates the changes in juvenile hormone III bisepoxide (JHB III) concentrations during the larval development of Chrysomya megacephala , Chrysomya rufifacies , and Musca domestica– three fly species of forensic importance in Southeast Asia and other tropical regions. JHB III was quantified using High-Performance Liquid Chromatography (HPLC). Pearson’s r correlation analysis examined relationships among larval weight, raw and normalized JHB III concentrations, and larval developmental age. Linear regression models were generated forsignificant correlations( p < 0.05) between normalized JHB III and larval developmental age. A fluctuating pattern of normalized JHB III concentrations throughout development was observed, with moderate negative correlations to larval age ( r = − 0.55 to − 0.67). These suggest that quantifying JHB III concentrations could supplement traditional morphological measurements like larval length, potentially increasing the accuracy and reliability of larval age estimates for mPMI calculation within the geographical range where these species are the forensic indicators.