Between 2005 and 2008, wild Bradypus variegatus and Choloepus hoffmanni inhabiting an agricultural landscape and captive animals from a rescue center in Northeastern Costa Rica were studied to assess exposure to pesticides. A total of 54 animals were sampled: 42 wild sloths captured at an agricultural landscape and 12 captive animals from a rescue center. Pesticides' active ingredients were determined in three sample matrices: hair, aqueous mixture (paws' wash) and cotton gauze (mouth clean) based on multi-residue gas chromatography methods. Recoveries tests ranged from 73 to 146% and relative standard deviations were less than 20% throughout all the recovery tests. Active ingredients detected in sloths samples were ametryn, chlorothalonil, chlorpyrifos, diazinon, difenoconazole, ethoprophos and thiabendazole. These active ingredients were used in intensive agricultural production for bananas, pineapples and other crops. Blood plasma cholinesterase activity (PChE) was determined by the Ellman method modified for micro plates. Enzyme activity determination was normalized to protein content in the samples according to Bradford method. Wild sloth PChE activity was similar for both species while sloths in captivity showed differences between species. Enzyme activity was significantly lower for two-toed sloths. This study showed that sloths were exposed to pesticides that caused acute and chronic effect in mammals and can also be a threat to other wildlife species. There is a need to better understand the potential effects of exposure to pesticides in sloths and other wild mammal populations, especially those threatened or endangered. More studies in this field must be carried out on the wildlife fauna inhabiting the agricultural landscape and its surroundings.

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