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Mercury is being released in the Amazon in an abusive way due to goldmining activities. The Tapajós river basin was the first to be intensively exploited in the modern Amazon gold rush. Fish and hair samples as the best indicators of human methylmercury contamination were investigated in the main cities and villages along the Tapajós river basin. The upper basin has typical fish fauna with much larger carnivorous fish with higher mercury levels reaching an average value of 0.69 μg·g⁻¹ wet wt. in 43 fish. This was accompanied by high levels in hair of the human population living in the same area. The maximum hair value reach 151 μg·g⁻¹ dry wt. with two villages presenting an average value close to 25 μg·g⁻¹ dry wt. An analytical laboratory intercalibration exercise was performed between Japanese and Brazilian laboratories for total mercury analysis. Critical fish, areas, and more exposed human groups are …