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.