Histopathology and transcriptome analysis reveals the gills injury and immunotoxicity in gibel carp following acute deltamethrin exposure

Abstract

More and more evidences proved that deltamethrin (Del) exposure induced adverse effects and damaged immune function to the aquatic animals in the parasite killing process with increasing insecticide application. However, little is currently known of the negative effect on mucosal immunity, especially in gills tissue. Therefore, this study was aimed to reveal the tissue injury and immunotoxicity in the gill of gibel carp following acute deltamethrin exposure. The LC50 of deltamethrin on gibel carp at 96 h was determined to be 6.194 μg/L, and then juvenile gibel carp (Carassius auratus gibelio) (8.8 ± 1.0 g) were exposed to four Del exposure groups (0.61, 1.22, 2.44, and 4.88 μg/L) for 12 h and 24 h. We measured the lysozyme (LYZ) contents and myeloperoxidase (MPO) activities and found that with increased concentration of Del exposure, the LYZ contents were found to increase in the 1.22 μg/L Del group initially significantly and then gradually significantly decrease in the 4.88 μg/L Del group. And the activities of MPO were significantly lifted in a dose-dependent manner. The histological analysis showed that Del exposure caused serious desquamation and necrosis in the surface of epithelial cells, accompanied by interlamellar cellular mass degenerative. In addition, the mucous cells were significantly decreased in the high Del concentration group (2.44 μg/L and 4.88 μg/L Del group) by AB-PAS staining. Additionally, totally 2857 DEGs (including 1624 up-regulated and 1233 down-regulated genes) were identified between the control group and 4.88 μg/L Del exposure group using transcriptional analysis. Among these, some genes involved in innate immune molecules, complement activation, apoptosis-related molecules, cytokine, and adaptive immune molecules, were also down-regulated. Importantly, we found immune system process and tumor necrosis factor receptor (superfamily) binding pathways were downregulated based on the GO and KEGG enrichment analysis. Meanwhile, we detected the expression of pro-inflammatory cytokines (TNF-α, IFN-γ, IL-1β, and IL-8), anti-inflammatory cytokines (TGF-β), LYZ, IgM, and Hsp70 in the gills tissue at 12 h and 24 h after Del exposure, which were consistent with our sequencing results. Collectively, these results demonstrated that the gills injury and immunotoxicity were induced by Del exposure and provided novel insight for explaining to some extent why Del-exposure fish are more susceptible to concurrent or secondary viral or bacterial infections.