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Abstract Background Evaluation of insect succession patterns is a scientific method to estimate the time elapsed since death. Several studies have shown that ante-mortem intake of opioids affect maggot growth rate. However, there are few published data that investigate the effect of ante-mortem opioid use on insect succession patterns. Therefore, the main purpose of this research was to investigate the effect of methadone on the succession patterns of insects on rabbit carcasses during the spring and winter of 2019. In the present study, the H-null hypothesis represents the dissimilarity between the successional waves of species from testing and control carcasses (H0: P: 0). Results During this study, 15 and 13 insect species were collected from carcasses during the spring and winter, respectively. The most dominant species during the both seasons were Chrysomya albiceps and Calliphora vicina. These two species preferred to lay eggs on the control carcasses earlier than the treated carcasses. Lucilia cuprina was observed only from the remains of untreated carcasses, while Saprinus chalcites and Necrobia rufipes were recorded only from the remains of treated rabbits. Samples indicate that 11.8% of the insects were members of the Coleoptera. Permutation analyzes based on the Mantel test were 0.647 ± 0.16 (P = 0.009) and 0.693 ± 0.16 (P = 0.003) for the similarity of the species between treated and untreated carcasses in the spring and winter, respectively. Permutation analyzes for the two most dominant fly species (Ch. albiceps and C. vicina) between the treated and untreated carcasses in the spring and winter were 0.515 ± 0.15 (P = 0.05) and 0.491 ± 0.14 (P = 0.09), respectively. Conclusion The results revealed that the overall pattern of insect succession was similar between the treated and untreated rabbit carcasses. However, the patterns of succession of Chrysomya albiceps and Calliphora vicina differed slightly between treated and untreated carcasses, and this could have an effect on the PMI min estimate.