Vultures are long-lived species sensitive to human-caused mortality that has already determined a widespread collapse in Asian and African populations. They provide significant ecosystem services (regulatory and cultural) consuming livestock carcasses and saving greenhouse gas emissions, favouring nutrient recycling, environmental sanitation, and providing financial revenue. Appraising the incidence and causes of mortality could help to improve management and conservation actions. We compiled records of reported mortalities for the reintroduced Griffon Vulture Gyps fulvus population of the central Apennines in Italy (123 cases, July 1994–December 2020). The average mortality was 4.69 vultures per year (± 1.14 SE), with no significant temporal trend. The peak of mortality events, estimated by harmonic regression analysis, was in March, while the minimum occurred in October. No differences were found among age classes and sex ratio mortality was established at 1.43:1 (M:F, N = 68). Out of 103 (83.7%) vultures which underwent a post-mortem and toxicological screening, 53% were poisoned, mainly by carbamates, and 27% died of unknown causes. Overall, direct or indirect anthropogenic mortality caused 67% of deaths. Even considering an inherent bias associated with reported mortality as to the prevalence of causes of death and estimation of mortality rates, the overwhelming relevance of poisoning highlights that existing anti-poisoning efforts should be refined and incorporated into a coordinated multidisciplinary strategy. A standardised approach, from vulture carcass discovery to post-mortem procedures and toxicological analysis, should be applied to reduce uncertainty in the determination of causes of death, increasing effectiveness in the prosecution of wildlife crimes. As most of the poisoning cases affecting the Griffon Vulture population in the central Apennines likely represent a side (though illegal) effect of retaliatory efforts to defeat livestock predators, effective strategies in reducing human–wildlife conflicts should be applied.