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Background Mercury is a global contaminant of concern though little is known about exposures in México. Objectives To characterize mercury levels in pregnant women, children, and commonly consumed seafood samples. Methods Use resources of the Early Life Exposures in Mexico to Environmental Toxicants (ELEMENT) birth cohorts to measure total mercury levels in archived samples from 348 pregnant women (blood from three trimesters and cord blood), 825 offspring (blood, hair, and urine) and their mothers (hair), and 91 seafood and canned tuna samples from Mexico City. Results Maternal blood mercury levels correlated across three trimesters and averaged 3.4 μg/L. Cord blood mercury averaged 4.7 μg/L and correlated with maternal blood from trimester 3 (but not trimesters 1 and 2). In children, blood, hair and urine mercury levels correlated and averaged 1.8 μg/L, 0.6 μg/g, and 0.9 μg/L, respectively. Hair mercury was 0.5 μg/g in mothers and correlated with child׳s hair. Mean consumption of canned tuna, fresh fish, canned sardine, and shellfish was 3.1, 2.2, 0.5, and 1.0 times per month respectively in pregnant women. Mean mercury content in 7 of 23 seafood species and 5 of 9 canned tuna brands purchased exceeded the U.S. EPA guidance value of 0.3 μg/g. Conclusions Mercury exposures in pregnant women and children from Mexico City, via biomarker studies, are generally 3–5 times greater than values reported in population surveys from the U.S., Canada, and elsewhere. In particular, mercury levels in 29–39% of the maternal participants exceeded the biomonitoring guideline associated with the U.S. EPA reference dose for mercury. [ABSTRACT FROM AUTHOR]