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Because of their persistence in aquatic environments and ability to impair reproduction and other critical physiological processes, organochlorine (OC) contaminants pose significant health risks to marine organisms. Despite such concerns, few studies have investigated levels of OC exposure in sharks, which are fish particularly threatened by anthropogenic pollution because of their tendency to bioaccumulate and biomagnify environmental contaminants. The present study examined concentrations of 29 OC pesticides and total polychlorinated biphenyls (PCBs) in the bonnethead shark (Sphyrna tiburo), an abundant species for which evidence of reproductive impairment has been observed in certain Florida populations. Quantifiable levels of PCBs and 22 OC pesticides were detected via gas chromatography and mass spectrometry in liver of 95 S. tiburo from four estuaries on Florida's Gulf coast: Apalachicola Bay, Tampa Bay, Florida Bay, and Charlotte Harbor. In general, OC concentrations were significantly higher in Apalachicola Bay, Tampa Bay, and Charlotte Harbor S. tiburo in relation to the Florida Bay population. Because the rate of infertility has been shown to be dramatically higher in Tampa Bay versus Florida Bay S. tiburo, the present findings allude to a possible relationship between OC exposure and reproductive health that requires further investigation. Pesticide and PCB concentrations did not appear to significantly increase with growth or age in S. tiburo, suggesting limited potential for OC bioaccumulation in this species compared with other sharks for which contaminant data are available. Concentrations of OCs in serum and muscle were not correlated with those in liver, indicating that these tissues are poor surrogates for measuring internal OC burden in this species via nonlethal sampling procedures.