Tramadol modulates Lucilia sericata development: clinical larval therapy and forensic insights

Entomotoxicology can inform both forensic reconstructions and clinical larval therapy, yet the consequences of patient-level drug exposure on therapeutic flies remain unexplored. We reared Lucilia sericata on bovine mince fortified with tramadol at therapeutic (0.05 mg/100 g) and lethal (2.0 mg/100 g) levels, alongside blank controls, and assessed development time, wing morphometrics, MALDI-TOF protein profiles, and targeted toxicology of adult tissues. Development time did not differ significantly among treatments, whereas morphometrics showed strong effects of treatment, sex, and their interaction: lethal exposure produced smaller wings, therapeutic exposure larger wings relative to controls, and females exceeded males across treatments. MALDI-TOF PCA primarily separated samples by tissue and experiment (batch), but adult leg spectra consistently distinguished lethal exposure from blank/therapeutic groups, indicating subtle treatment-linked molecular variation. LC-MS/MS detected tramadol in adults exposed to lethal tramadol concentrations in two of three experiments while O-desmethyltramadol was not detected; both analytes were undetectable at therapeutic levels. Collectively, tramadol induced pronounced morphological shifts without measurable developmental delay, a combination that could bias PMI_min estimates in forensic entomology and, in clinical settings, possibly influence maggot therapy performance. These findings support integrating morphometrics with targeted proteomics and toxicology in entomotoxicological evaluations.