We studied white-nosed coatis (Nasua narica) in Monteverde, Costa Rica to evaluate their potential as a reservoir for the vector-borne zoonotic parasite Trypanosoma cruzi and other selected microorganisms. We live-trapped 20 coatis in July and August 2011 and tested them for T. cruzi by blood smear, molecular analysis of blood, culture of blood and anal gland secretions, and serology. Seven coatis (35%) were polymerase-chain-reaction–positive for T. cruzi and one coati was also culture positive. We did not detect T. cruzi in anal gland secretions. All coatis were positive for Mycoplasma and Babesia, but were negative for Baylisascaris, Anaplasma, Candidatus Neoehrlichia lotoris, Ehrlichia, Bartonella, and several apicomplexan parasites. The possible pathogenicity of T. cruzi, Babesia, and Mycoplasma in coatis and their transmission potential to humans and domestic animals warrants further investigation.