Mercury (Hg) contamination in fish has raised global concerns for decades. The Hg biotransformation can be manipulated by gut microbiome and it is found to have a substantial impact on the speciation and final fate of Hg in fish. However, the contribution of intestinal microbiota in geographical and interspecies variations in fish Hg levels has not been thoroughly understood. The present study compared the Hg levels in wild marine fish captured from two distinct regions in South China sea. We observed a quite “ironic” phenomenon that MeHg levels in carnivorous fish from a region with minimal human impacts (Xisha Islands, 92 ± 7.2 ng g−1 FW) were much higher than those from a region with severe human impacts (Daya Bay, 19 ± 0.41 ng g−1 FW). Furthermore, the results showed that gut microbiome determined Hg biotransformation and played a crucial role in the variances in fish Hg levels across different geographical locations and species. The intestinal methylators, rather than demethylators, were more significant in affecting Hg biotransformation in fish. The carnivorous species in Xisha Islands exhibited a higher abundance of intestinal methylators, leading to higher MeHg accumulation. Besides, the gut microbiome could be shaped in response to the elevated Hg levels in these fish, which may benefit their adaptation to Hg toxicity and overall health preservation. However, anthropogenic activities (particularly overfishing) in Daya Bay have severely affected the fish population, disrupting the reciprocal relationships between fish and intestinal microbiota and rendering them more susceptible to pathogenic microbes. Overall, this study provided a comprehensive understanding of the role of gut microbiome in Hg bioaccumulation in fish and offered valuable insights into the co-evolutionary dynamics between fish and gut microbiome in the presence of Hg exposure.