In vitro immune function in laboratory-reared age-0 smallmouth bass (Micropterus dolomieu) relative to diet

Smallmouth bass (Micropterus dolomieu) are used as an indicator species in environmental monitoring and assessment studies. However, laboratory-based studies for methods development and effector assessment are limited for this species. Nutrition, a known modulator of teleost physiological responses including immune function, is a critical knowledge-gap sometimes overlooked in the design of laboratory studies. We report the results from a study evaluating a commercially available artificial pelleted diet for bass and live feed (fathead minnows). Following a six-month diet-acclimation period, age-0 smallmouth bass were assessed using morphometric measures, histologic and immune-function end points using conventional methods, miniaturized cell isolation and assay methods as well as imaging flow cytometry. Fish on the two diets did not significantly differ in length, weight, or condition factor, indicating that growth was similar in the two groups. Histologic examination revealed relatively higher levels of macrophage aggregates and accumulation of ceroid/lipofuscin in the spleen as well as hepatocyte changes in the pellet-fed group. Leukocytes from the pellet-fed group exhibited significantly elevated bactericidal activity and significantly depressed mitogen response compared to fish fed live feed. Following exposure to a known inducer of inflammatory responses, bacterial lipopolysaccharide, responses including the transition of leukocytes to an apoptotic/necrotic condition differed significantly based on diet. Histologic findings were consistent with the occurrence of diet-related oxidative stress in the pellet-fed fish. Oxidative stress can be induced by multiple factors including environmental pollutants. For a diet to be useful in laboratory-based studies, it cannot elicit response that could also be induced by experimental treatments. To do so greatly complicates the detection of experimental effects. Until an artificial diet is developed for smallmouth bass that does not produce potentially confounding conditions for laboratory-based studies, use of a live feed appears to be the best option.