Trypanosoma cruzi is an important cause of disease and death in humans and dogs, and although wildlife infections are common, less is known about disease manifestations. A 12-week-old male American black bear (Ursus americanus) cub with mild lethargy and anorexia presented to a wildlife rehabilitation center in Lake Tahoe, California. The cub continued to become increasingly weak and showed decreasing interest in play and other activities. The cub was anemic and had increased γ-glutamyltransferase (GGT) liver enzymes. A large number of trypanosomes were noted on a thin blood smear. Trypanosoma cruzi was isolated in culture from a subsequent blood collection. Proliferative bony lesions were noted on radiographs, but this finding was considered unrelated to the T. cruzi infection. The number of parasites observed in thin blood smears dramatically dropped over time, but it remained PCR positive until at least nine months. The cub continued to gain weight and became increasingly active. Serum samples from the cub were positive with three different serologic assays (IFA, ELISA, and ICT). The bear was not treated because of the decreasing parasitemia and the improvement in activity and appetite. Although the bear could not be released due to issues unrelated to T. cruzi, it remains healthy in a captive facility. Sequence analysis of the DHFR-TS and COII-ND1 gene sequences confirmed the bear was infected with DTC TcIV. Following the detection of this clinical case, a serologic survey was conducted to determine the prevalence of T. cruzi exposure of black bears in California, North Carolina, and Pennsylvania. Because no serologic assay has been validated for use in bears, three different assays were used. Marked differences in apparent seroprevalence range from 1% (requiring all three assays to be positive) to ∼20.7% (requiring only one assay to be positive). Black bears are naturally exposed to T. cruzi across the United States. Future studies using PCR testing of tissues or blood would be needed to better understand the prevalence of T. cruzi in wild black bears, lineages most commonly associated with infection, and if T. cruzi represents a health threat to bears.