Hepatic Gene Expression Changes of Zebrafish Fed Yeast Prebiotic, Yeast Probiotic, Black Soldier Fly Meal, and Butyrate

Abstract

As global fish consumption rises, improving fish health through immunomodulatory feed ingredients shows promise while also supporting growth performance. This study investigated the effects of yeast prebiotics, probiotics, a postbiotic (butyrate), and black soldier fly larvae (BSFL) meal on fish immune responses. Zebrafish were fed diets containing these ingredients for 63 days and then exposed to either Pseudomonas aeruginosa lipopolysaccharide (LPS) or live Flavobacterium psychrophilum to assess hepatic candidate gene expression and weight gain. No mortalities were observed post-immune challenges, and weight gains were not significantly different across treatments. Liver samples were collected for mRNA analysis, and real-time qPCR was used to evaluate the expression of immune-related genes such as TNF-ɑ, IL-1β, hepcidin, and NF-κB/p65. NF-κB/p65 was upregulated in response to immune challenges, indicating a reaction to both LPS and pathogen exposure. Fish on the BSFL diet showed decreased NF-κB/p65 expression after the pathogen challenge, while probiotic-fed fish had reduced angiopoietin-like 4 (angptl4) levels following LPS exposure. Butyrate supplementation had the most significant impact, downregulating pro-inflammatory cytokines and other immune-related genes, suggesting a protective effect. These findings support the health benefits of BSFL and sodium butyrate during an immune challenge.