Difenoconazole causes spleen tissue damage and immune dysfunction of carp through oxidative stress and apoptosis

As the use of pesticides increases year after year, so does the level of residual pesticides in the aquatic environment, posing a serious threat to non-target organisms. Difenoconazole (DFZ), a class of long-lasting fungicides and residues in the marine environment, has been shown to cause damaging effects on different organs of aquatic organisms. However, there is no research on the damage of DFZ to carp spleen tissue. This study aimed to investigate the acute toxic effects of DFZ on the spleen tissue of carp (Cyprinus carpio) by exposing juvenile carp to environmentally relevant concentrations of DFZ. We randomly selected 30 carp, divided them into the Control, Low, and High groups, and then exposed the three groups to 0, 0.488 mg/L DFZ, and 1.953 mg/L DFZ for 96 h respectively. We then investigated the toxic effects caused by DFZ on carp and spleen tissues by detecting changes in spleen histopathologic damage, apoptosis, oxidative stress, inflammation, and blood biochemical parameters. We found that DFZ causes severe histopathology in spleen tissue, including ballooning, structural relaxation, and giant mitochondria. In addition, we found that DFZ caused excessive apoptosis in spleen tissue by TUNEL staining and expression levels of apoptosis-related genes (caspase3, caspase8, caspase9, fas, bax, bcl-2, and p53). The activities and transcript levels of the antioxidant enzymes SOD, CAT, and GSH-Px were significantly down-regulated. In addition, DFZ led to a significant increase in activation of the NF-kappa B signaling pathway and mRNA levels of pro-inflammatory cytokines il-6, il-1 beta, and tnf-alpha, and a substantial decrease in mRNA levels of anti-inflammatory cytokines il-10 and tgf-beta 1 in spleen tissue. Blood biochemical parameters showed that DFZ exposure significantly reduced erythrocyte, leukocyte, hemoglobin, C3, and IgM levels. Collectively, DFZ exposure induced apoptosis, immunosuppression, oxidative stress, and inflammatory responses in the spleen tissue of carp, resulting in spleen tissue damage.

Automated summary

With the increasing use of pesticides, there is a growing concern about the levels of residual pesticides in the aquatic environment and their impact on non-target organisms. This study focused on the acute toxic effects of difenoconazole (DFZ), a class of long-lasting fungicides, on the spleen tissue of carp. The results showed that DFZ caused severe histopathology, apoptosis, oxidative stress, and inflammation in the spleen tissue of carp, leading to tissue damage. These findings highlight the importance of ensuring the safety of aquatic organisms in pesticide usage.