The West Indian manatee (Trichechus manatus) is exposed to a number of anthropogenic influences, including metals, as they inhabit shallow waters with close proximity to shore. While maintaining homeostasis of many metals is crucial for health, there is currently no baseline reference range that can be used to make clinical and environmental decisions for this endangered species. In this study, whole blood samples from 151 manatees were collected during health assessments performed in Florida and Belize from 2008 through 2011. Whole blood samples (n = 37) from managed care facilities in Florida and Belize from 2009 through 2011 were also used in this study. The concentrations of 17 metals in whole blood were determined, and the data were used to derive a baseline reference range. Impacts of capture location, age, and sex on whole blood metal concentrations were examined. Location and age were related to copper concentrations as values were significantly higher in habitats near urban areas and in calves. Copper may also be a husbandry concern as concentrations were significantly higher in managed manatees (1.17 ± 0.04 ppm) than wild manatees (0.73 ± 0.02 ppm). Zinc (11.20 ± 0.30 ppm) was of special interest as normal concentrations were two to five times higher than other marine mammal species. Arsenic concentrations were higher in Belize (0.43 ± 0.07 ppm), with Placencia Lagoon having twice the concentration of Belize City and Southern Lagoon. Selenium concentrations were lower (0.18 ± 0.09 ppm) than in other marine mammal species. The lowest selenium concentrations were observed in rehabilitating and managed manatees which may warrant additional monitoring in managed care facilities. The established preliminary baseline reference range can be used by clinicians, biologists, and managers to monitor the health of West Indian manatees.