High temperature exposure causes sexual dimorphism in immune responses against largemouth bass ranavirus (LMBV) in Micropterus salmoides

Largemouth bass (Micropterus salmoides) is an important freshwater fish species in aquaculture. Largemouth bass ranavirus (LMBV) infection at high temperature results in a considerably higher mortality rate in females than that in males. This study compared the spleen transcriptomes and analyzed the impact of sexual dimorphism on immunity and metabolism of largemouth bass infected with LMBV at high temperatures to gain a comprehensive understanding of the sex-based differences in immunity and metabolism of this species. (1) Compared with the ambient temperature group (W20), the high temperature group (W30) exhibited multiple significant differences in metabolic and immune terms, with several immunity-related pathways, including phosphatidylinositol 3-kinase-protein kinase B (PI3K-Akt) signaling pathway. Most identified pathways were associated with the estrogen signaling pathway, which also differed significantly. (2) Differences in glycolipid metabolism, immunity, and the endocrine and complex adaptive systems were distinctly different between male groups (W20M vs. W30M), whereas the female groups (W20F vs. W30F) were not significantly different. The male groups had a significantly higher number of enriched immunity-related KEGG pathways and differentially expressed genes (DEGs) and pathways related to the estrogen signaling pathway. (3) When comparing male and female samples infected with LMBV at the same temperature, no significant difference was observed between the ambient temperature groups (W20F vs. W20M). The high-temperature groups (W30F vs. W30M) had 18 immunity-related pathways among the top 25 enriched KEGG pathways, and gene expression correlation analysis of the DEGs showed positive correlations. This study demonstrates that the combined effects of high temperature and LMBV affect multiple metabolic and immune pathways in largemouth bass, thereby modulating immune responses by regulating the estrogen signaling pathway and influencing the expression of estrogen and its receptors. Under high-temperature conditions, male fish may have stronger estrogen signaling pathway-associated immune responses, which regulate immune pathways, including antigen processing and presentation, complement system, and phagocytosis. Consequently, males may have higher disease resistance than that of females. Our result contribute to mono-sex breeding and antiviral drug therapies available in largemouth bass, thereby markedly improving breeding efficiency.