Differentially expressed genes involved in immune pathways following Vibro parahaemolyticus challenge: A study on red swamp crayfish Procambarus clarkii

Abstract

This study conducted a comprehensive analysis of the damage mechanism of Vibrio parahaemolyticus (V. parahaemolyticus) on crayfish, utilizing pathological section observations, biochemical indicator detection, and transcriptomics analysis. Histopathological examination showed fragmentation of the gill lamellae, spotted pigment deposition on the branchial membrane, degeneration of epithelial cells, and disruption of the normal structure of gill tissues. Biochemical tests indicated a significant decrease in the activity of antioxidant-related enzymes in crayfish injected with V. parahaemolyticus compared with the phosphate-buffered saline group. Transcriptomics analysis identified 2307 upregulated and 1721 downregulated differentially expressed genes (DEGs). Gene ontology enrichment analysis highlighted that DEGs were primarily involved in the regulation of lipid metabolic processes, cellular nitrogen compound biosynthesis and metabolic processes, and the negative regulation of neuron death. Kyoto Encyclopedia of Genes and Genomes pathway analysis revealed significant enrichment in N-glycan biosynthesis, fatty acid metabolism, and the mTOR signaling pathway. Gene set enrichment analysis indicated pathways related to immunity and oxidative stress, including oxidative phosphorylation and tyrosine metabolism. Quantitative reverse transcription–polymerase chain reaction results for selected genes involved in the oxidative phosphorylation pathway and immune-related genes corroborated the RNA-Seq trends, confirming the transcriptomics data's reliability. The findings suggest that infection with V. parahaemolyticus leads to excessive oxidative stress due to bacterial proliferation, overwhelming the crayfish's antioxidative defense. This causes a continuous increase in reactive oxygen species levels, leading to irreversible damage to the oxidative phosphorylation and innate immune systems. This study elucidated the mechanism of oxidative stress damage in crayfish following V. parahaemolyticus infection, providing valuable insights for reducing bacterial and viral diseases in aquaculture. A deeper understanding of pathogen affects the physiology and immune systems of aquatic organisms can help develop more effective prevention and control strategies, thereby improving the efficiency and sustainability of aquaculture.