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Poly-β-hydroxybutyrate (PHB) can promote growth and enhance immunity of aquatic animals such as gibel carp, Carassius auratus gibelio. However, the optimal dietary supplementation level and feeding duration for gibel carp are still unclear. In this study, effects of various PHB supplementation levels on gibel carp growth performance, digestive enzyme activities, immune function, disease resistance, and intestinal mucosal barrier were evaluated. Six experimental diets differing in PHB levels were formulated, that is, 0% (P0 as control), 0.5% (P0.5), 1% (P1), 2% (P2), 4% (P4), and 8% (P8). Results indicated that PHB promoted fish growth performance significantly (p < 0.05). Compared to control group P0, the intestinal lipase, protease, and amylase activities in group P4 were significantly increased after 30 days (p < 0.05). Meanwhile, dietary PHB supplementation at 4% level for 30 days and 0.5% for 60 days upregulated the mRNA expression of immune-related

Cyprinid herpesvirus 2 (CyHV-2) infects gibel carp (Carassius auratus gibelio) and causes severe losses. Microbiota in animal guts involves nutrition intake, development, immunity, and disease resistance. However, the relationship between gibel carp gut microbiota and CyHV-2 infection is not well known. Herein, we analyzed the gut microbiota composition and metabolite profiles in CyHV-2-infected and -uninfected fish using high-throughput sequencing and gas chromatography/mass spectrometry. Results showed that CyHV-2 infection significantly changed gut microbiota and metabolite profiles (p &lt; 0.05). High-throughput sequencing demonstrated that the relative abundance of Aeromonas in the midgut increased dramatically while Cetobacterium decreased. Time-course analysis showed that the number of Aeromonas in the midgut of infected fish increased more than 1,000 times within 5 days post infection. Metabolome analysis illustrated that CyHV-2 infection significantly altered 24 metabolites in the midgut of gibel carp, annotating to the anomaly of digestion and metabolisms of amino acids, carbohydrates, and lipids, such as tryptophan (Trp) metabolism. The Mantel test demonstrated that gut microbiota and metabolite profiles were well related (r = 0.89). Furthermore, Trp metabolism responded to CyHV-2 infection closely was taken as one example to prove the correlation among CyHV-2 infection, metabolites and microbiota in the midgut, and host immunity. Results showed that modulating Trp metabolism could affect the relative abundance of Aeromonas in the midgut of fish, transcription of antiviral cytokines, and CyHV-2 infection. Therefore, we can conclude that CyHV-2 infection significantly perturbed the gut microbiome, disrupted its’ metabolic functions, and caused the proliferation of the opportunistic pathogen Aeromonas. This study also suggests that modulation of the gut microbiome will open a therapeutic opportunity to control CyHV-2 infection in gibel carp.