Clinico-pathological findings and expression of inflammatory cytokines, apoptosis, and oxidative stress-related genes draw mechanistic insights in Nile tilapia reared under ammonia-N exposure and Aeromonas hydrophila challenge

Abstract

Fish diseases have a “stress-related” nature, whereas fish exposure to stressors will increase their susceptibility to infections. It was also noted that fish exposure to biotic and abiotic stressors would exaggerate the disease signs, elicit high mortalities, and cause severe economic losses. Motile aeromonad septicemia (MAS) is a major bacterial disease affecting a variety of finfish species throughout the globe and is caused by Aeromonas hydrophila. Herein, we have evaluated the impacts of ammonia-N stress and/or Nile tilapia challenge with pathogenic A. hydrophila on the clinical picture of MAS disease. Clinical signs, postmortem lesions, histoarchitectural changes, and gene transcription analysis were studied. Fish experimentally infected with A. hydrophila were exophthalmic and showed darkened skin. Moreover, opercular hyperemia, petechial hemorrhages, and gill congestion alongside dermal ulcerations were noticed in ammonia-exposed fish. On the other side, fish exposed to both stressors exhibited exophthalmia, corneal opacity, severe dropsy, and hemorrhagic dermal ulcerations. At the tissue levels, the histopathological lesions were exaggerated in the fish group exposed to ammonia stress and challenged with A. hydrophila than fish group exposed to each one alone. At the molecular levels, the mRNA expression analysis reveals significant upregulation of inflammatory cytokines such as interleukin-1 beta, CXC chemokine, and tumor necrosis factor-alpha in the kidney tissues of Nile tilapia exposed to ammonia and/or challenged with A. hydrophila. In a similar trend, the mRNA expression values of heat shock protein 70 (HSP70), oxidative stress related genes (SOD and CAT) and apoptosis-related genes (caspase 3, BAX, and cytochrome P450) were also increased in the hepatic tissues of fish exposed to singular or dual stressors. Interestingly, the highest expression levels of the above-mentioned genes were found in the fish group exposed to both stressors. Taken together, these findings indicate the occurrence of severe inflammatory and apoptotic changes in fish exposed to ammonia and infected with A. hydrophila more than each one alone. In contrast, there was a significant decrease in the expression values of the antioxidant enzyme glutathione-S-transferase (GST) in stressed fish, suggesting the occurrence of oxidative stress. This study will be helpful to draw mechanistic insights into the exposure of fish to ammonia stress and infection with A. hydrophila.