Effects of bacterial haemorrhagic septicemia on the immune response of Leiocassis longirostris by RNA-Seq and microRNA-Seq

Leiocassis longirostris is a common fish variety that is widely cultivated in China, during the breeding process however, it is highly susceptible to bacterial haemorrhagic septicemia, which can cause great economic loss for farmers. To understand the immune responses of L. longirostris to Aeromonas hydrophila infection, Illumina sequencing was employed to identify changes in the mRNA and miRNA in spleen tissue. In this study, a total of 92.16 and 95.61 million (M) high-quality transcriptome reads were generated from the control group (CG) and experimental group (EG) spleen samples, respectively, and 207 up-regulated and 185 down-regulated genes were identified. These genes were enriched in 29 GO terms and 30 KEGG pathways (P ≤ 0.05), including cytokine-cytokine receptor interaction and complement and coagulation cascades, with 17 up-regulated genes and 12 down-regulated genes related to immune responses in the EG relative to the CG. Based on the zebrafish genome, miRNA-seq identified a total of 343 miRNAs, of which 15 were up-regulated and 10 were down-regulated (fold-change ≥2 or ≤0.5 and P ≤ 0.05). Target gene prediction and KEGG enrichment analysis revealed that all of the target genes were concentrated in 13 pathways associated with immune response, including the mTOR signaling pathway and the TGF-beta signaling pathway. The expression patterns of 8 differentially expressed genes and 4 miRNAs involved in immune response were validated by quantitative real-time RT-PCR. These results have provided valuable insights into the molecular mechanisms underlying the immune response of L. longirostris to bacterial haemorrhagic septicemia.