Health evaluations of brown pelican (Pelecanus occidentalis) nestlings from three colonies along the Atlantic coast of the southeastern United States were performed in 2005, 2007, and 2008. The primary objective of this study was to establish baseline data for hematologic, biochemical, and serologic values from a relatively healthy population of free-living pelicans during early chick development. Relationships among health variables and colony site, ectoparasite infestation, sex, and body condition index were also evaluated. Reference intervals are presented for health variables, including novel analytes for the species, as well as a comparison of these results with previously published values for wild pelicans. No significant relationships were found between health variables and nestling sex or body condition; however, differences between colony sites and the presence of ectoparasites were detected. The inclusion of health assessments as a regular component of management programs for seabirds can provide data to better understand the effect to species of concern when drastic changes occur to the population and its environment.