The health of resident fishes serves as a biologically relevant barometer of aquatic ecosystem integrity. Here, the health of the Ottawa River and Ashtabula River (both within the Lake Erie Basin) were assessed using morphological and immunological biomarkers in brown bullheads (Ameiurus nebulosus) and largemouth bass (Micropterus salmoides). Biomarker metrics were compared to fish collected from a reference site (Conneaut Creek). Data utilized for analyses were collected between 2003 and 2011. Fish collected from all three river systems had markedly different contaminant profiles. Total PCBs were the dominant contaminant class by mass. In bullhead, PCBs were highest in fish from the Ashtabula River and there were no differences in fish collected pre- or post-remediation of Ashtabula Harbor (median = 4.6 and 5.5 mg/kg respectively). Excluding PCBs, the Ottawa River was dominated by organochlorine pesticides. Liver tumor prevalence exceeded the 5% trigger level at both the Ashtabula (7.7%) and Ottawa Rivers (10.2%), but was not statistically different than that at the reference site. There was no statistically significant association between microscopic lesions, gross pathology and contaminant body burdens. Collectively, contaminant body burdens were generally negatively correlated with functional immune responses including bactericidal, cytotoxic-cell and respiratory burst activity in both species. Exceptions were positive correlations of HCB and heptachlor epoxide with respiratory burst activity in largemouth bass, and HCB with respiratory burst activity in bullhead and ΣBHC for all three functional assays in bullhead. Data here provide additional support that organochlorine contamination is associated with immunomodulation, and that species differences exist within sites.