Development of a suite of functional immune assays and initial assessment of their utility in wild smallmouth bass health assessments

Methods were developed for measuring immune function in Micropterus dolomieu (smallmouth bass). The ultimate objective is to monitor and evaluate changes over time in immune status and disease resistance in conjunction with other characteristics of fish health and environmental stressors. To test these methods for utility in ecotoxicological studies, 192 smallmouth bass, age 2 years and older, were collected from three sites within the Susquehanna River Basin and one site in the Ohio River Basin during spring and fall 2016 and 2017. The anterior kidney was aseptically removed and homogenized for leukocyte isolation. Leukocytes were tested for bactericidal activity against two species of bacteria; respiratory burst activity when stimulated with phorbol 12-myristate 13-acetate; and mitogenesis activity when stimulated with concanavalin A, phytohemagglutinin, and lipopolysaccharide. Tissues were preserved for histopathological analyses.

Two of the sites were part of a monitoring program at which surface-water samples were collected monthly (bimonthly in spring) for chemical contaminants. Significant seasonal and (or) site differences in all three immune function tests were observed. Interpretations of seasonal trends in immune function of wild fish or correlations with environmental variables and other factors are difficult to make owing to the complex nature of the immune response and the environment. Differences in immune function could potentially be related to a variety of confounding factors; therefore, additional endpoints and repeated sampling over an extended period are essential to draw conclusions on the immune status of wild fish.