Acute exposure to chlorpyrifos induces reversible changes in health parameters of Nile tilapia (Oreochromis niloticus)

Abstract

Chlorpyrifos (CPF) is one of the most common insecticides found in freshwater ecosystems, and has been detected in agricultural and fishery products worldwide. This study focused on comprehensive panel of hematological, immunotoxic and pathology changes in Nile tilapia (Oreochromis niloticus) during and after exposure to CPF at 15 μg/L (0.043 μM) (1/10 LC₅₀, group CPF1), or 75 μg/L (0.21 μM) (1/2 LC₅₀, group CPF2) for 14 days, followed by 2 weeks recovery. Different endpoints were used to determine effects of CPF on fish health: hematological parameters; antioxidant levels in liver and gills; innate immune parameters; expression levels of pro-inflammatory cytokine genes at mRNA level in anterior kidney and spleen; and histopathological assessment of gills, liver, and kidney tissues. RBCs were significantly decreased in CPF1 group compared to other groups only at day 3. Blood packed cell volume (PCV) and mean corpuscular volume (MCV) showed significant increase at day 3 and 14 of CPF exposure. TLC (Total Leukocytic Counts), neutrophil counts were significantly increased in CPF exposed groups at days 3, 7, 14 compared to the control. While, lymphocytes counts were significantly increased at CPF1 group compared to other groups at day 14. Antioxidant enzyme activity in liver and gills showed significant increase of Malondialdehyde (MDA) and glutathione (GSH), and significant decrease in (catalase/CAT/, glutathione S-transferase/GST/, and superoxide dismutase/SOD/); in CPF exposed groups. Serum bactericidal and lysozyme activity was nominally and significantly decreased, respectively, and whole blood respiratory burst was significantly increased in CPF2 group. The cytokine expression levels showed complex changes in expression patterns. In kidney, cytokine interleukin-8 (IL-8) was significantly upregulated at day 1 in both exposed group. Interleukin-1β (IL-1β) and Tumor necrosis factor-α (TNFα) were significantly upregulated at day 1 in CPF1 group, and then IL-8 and TNFα downregulated at day 3 in same group. At day 7, only TNFα was up and downregulated in CPF1 and CPF2, respectively compared to control. All gene expression levels in spleen were upregulated on day 7 of exposure in the high exposed group. Histopathology showed dose-dependent changes in CPF treated groups, indicating gill, liver, and posterior kidney changes associated with oxidative stress damages. Following recovery period, all measured parameters showed varying degrees in their reversibility to the control level. These findings provide important insights about the acute toxic effects of CPF on fish and show potential to be used as biomarkers in further toxicological evaluation studies.

Keywords: Fish health; Hematology; Innate immunity; Oxidative damage; Pesticides; Pro-inflammatory cytokines.