The Analysis of Goldfish (Carassius auratus L.) Innate Immune Responses After Acute and Subchronic Exposures to Oil Sands Process-Affected Water

We examined the immunotoxic effects of acute and subchronic exposures of goldfish to aged, fresh, and ozonated oil sands process-affected water (OSPW) using a flow-through exposure apparatus. We measured the expression of proinflammatory cytokine genes, the antimicrobial responses of primary macrophages isolated from OSPW-exposed fish, and the ability of the goldfish to control infection with a protozoan parasite, Trypanosoma carassii. After acute (1 week) exposure to aged OSPW, we observed upregulation in the expression of interferon gamma (IFN-), tumor necrosis factor alpha-2 (TNF-2) in the kidney and spleen but not in gills of the fish. After subchronic (12 weeks) exposure to aged OSPW, we observed significant increases in mRNA levels of proinflammatory genes in the gill (IFN-, interleukin-1 beta 1 [IL1-1], TNF-2), kidney (IL1-1, TNF-2), and spleen (IL1-1). An upregulation of immune gene expression in the gill and kidney (IFN-, IL1-1, TNF-2) and spleen (IL1-1, TNF-2) was observed after acute exposure of fish to diluted fresh OSPW. Following subchronic exposure to diluted fresh OSPW, we observed high mRNA levels of IL1-1 in all tissues examined. However, there were significant decreases in the mRNA levels of IFN- and TNF-2 in the kidney and spleen and gill and spleen (IL-12p35 and IL-12p40) of exposed fish. There were no changes in the expression of anti-inflammatory cytokine IL-10 after both acute and subchronic exposures to diluted fresh OSPW. In fish exposed to ozonated fresh OSPW, immune gene expression was similar to nonexposed control fish in all organs examined, with exception of IL1-1. The ability of primary kidney macrophages to generate reactive oxygen and nitrogen intermediates was significantly reduced in fish exposed to fresh OSPW. The enhanced proinflammatory response after acute exposure to diluted fresh OSPW was confirmed by the parasite challenge experiments, where OSPW-exposed fish controlled the infection better than nonexposed fish.