Mercury (Hg) is a chemical element that poses risks to human health due to its high toxicity and environmental persistence. We determined the total Hg (THg) and methyl Hg (MeHg) concentrations in hair samples from residents of the Demarcação District (Porto Velho, Rondônia) in the Brazilian Amazon, as well as in water and fish samples, to evaluate factors influencing human exposure. The average THg concentration in human hair was 7.86 ± 6.78 mg kg−1 and it was significantly higher in men, with an increasing trend related to age. There was no significant difference between female age groups. Human exposure to Hg through water was negligible compared to fish consumption. The average weekly intake estimates in the community varied between 1.54 and 4.62 μg kg−1, substantially higher than the recommended limit. The fish species with the highest amounts safe for daily consumption were herbivores and detritivores. Our results contribute to an understanding of how exposure to Hg affects the health of riverside populations and provide insights for new research to develop methods to mitigate such exposure and thus improve the quality of life of Amazonian people.