Terrestrial Compound Protein Replacing Dietary Fishmeal Improved Digestive Enzyme Activity, Immune Response, Intestinal Microflora Composition, and Protein Metabolism of Golden Pompano (Trachinotus ovatus)

Abstract

Terrestrial compound protein (Cpro) can be potentially used to replace fishmeal (FM) in the marine carnivorous teleost, golden pompano (Trachinotus ovatus). Four isonitrogenous (45%) and isolipidic (12%) diets named FM30, AP80, PP80, and CP80 were formulated. FM30 (control) contained 30% FM and 25% basic protein, while AP80, PP80, and CP80 only contained 6% FM, where 80% FM and 25% basic protein of control diet were completely replaced by animal protein, plant protein, and Cpro, respectively. After golden pompano juveniles (initial weight: 10.32 ± 0.09 g) were, respectively, fed the four diets in floating sea cages for 10 weeks, the growth performance, intestinal digestive enzyme activity, and immune responses, protein metabolism indices of the CP80 group were similar to or better than those of the FM30 group (P > 0.05), and significantly better than those of the AP80 and PP80 groups. Specifically, the weight gain (WG), feed conversion ratio (FCR), activity of alanine transaminase (ALT), growth hormone (GH), and insulin-like growth factor-1 (IGF-1) contents of serum, mRNA level of interleukin-10 (il-10), zonula occludens-2 (zo-2), claudin-3, claudin-12, and eukaryotic translation initiation factor 4G (eif4g) were significantly higher, and the activity of α-amylase (AMS), lipase (LPS) in the foregut and midgut, interleukin-8 (il-8) expression in the intestine was significantly lower than that in the CP80 group, compared with those in AP80 and PP80 groups (P < 0.05). Moreover, the intestinal microflora composition of golden pompano fed with the CP80 diet was improved. Specifically, at the phylum level, the relative abundance of harmful bacterial strains cyanobacteria and TM7 of CP80 group was similar to those of FM30 group (P > 0.05), but was significantly lower than those of AP80 and PP80 groups (P < 0.05). At the genus level, the beneficial bacterial strains Agrobacterium and Blantia of CP80 group were also similar to those of FM30 group (P < 0.05), which were significantly higher than those of AP80 and PP80 groups, but the beneficial bacterial strains Bifidobacterium and Devosia of CP80 group were significantly higher than that in the other groups (P < 0.05). Besides, in diet CP80, the contents of amino acids and anti-nutritional factor, as well as the in vitro digestion rate were comparable to those of FM30, and the anti-nutritional factor content was between AP80 and PP80; total essential amino acids (EAAs) and methionine contents were higher than those in AP80, the glycine content was higher than that in PP80. Taken together, these results indicated that the CP80 diet had better amino acid composition and relatively low content of anti-nutritional factors, as well as high-digestion rate, and thus leads to the fish fed CP80 displaying improved effects in digestive enzyme activity, immune response, protein metabolism, and intestinal microbiota composition, which may be the important reasons to explain why that 80% of FM can be replaced by Cpro in the diet of golden pompano.

Figure 1

β Diversity of intestinal microbiota of juvenile golden pompano at the phylum (a) and genus (b) levels principal component analysis (PCA) against PC1 versus PC2 axes based on OTUs.

Figure 2

Heatmap of the abundance of golden pompano intestinal bacteria at phylum (a) and genus (b) levels in different experiment groups. Phylogenetic positions are projected by the OTUs, and the taxa of OTUs are listed on the right. Color intensity indicates the relative enrichment of OTUs.

Figure 3

Intestinal bacterial composition at the phylum (a) and genus (b) levels in different experiment groups. Top 10 most abundant bacterial phyla and genera were shown in the figures, respectively. Other phyla and genera were all assigned as “Others”.

Figure 4

Phylum (a) and genus (b) level relative abundances of intestinal flora of juvenile golden pompano fed different diets for 10 weeks. Values presented as means ± SEM (n = 6). Bars assigned by different letters are significantly differed at P < 0.05.

Figure 5

Relative mRNA expression of inflammation-related genes (a) and tight junction protein genes (b) in the intestine juvenile golden pompano fed different diets for 10 weeks. Values presented as means ± SEM (n = 6). Bars assigned by different letters are significantly differed at P < 0.05. il-8, interleukin-8; tnf-α, tumor necrosis factor-α; il-10, interleukin-10; tgf-β1, transforming growth factor-β1; zo-1, zonula occludens-1; zo-2, zonula occludens-2.

Figure 6

Serum growth hormone (μg/L) and insulin-like growth factor-1 (μg/L) content of juvenile golden pompano fed different diets for 10 weeks. Values presented as means ± SEM (n = 6). Bars assigned by different letters are significantly differed at P < 0.05.

Figure 7

Relative expression of the mammalian target of rapamycin (mtor), ribosomal protein S6 kinase 1 (s6k1), and eukaryotic translation initiation factor 4G (eif4g) in the liver of juvenile golden pompano fed different diets for 10 weeks. Values presented as means ± SEM (n = 6). Bars assigned by different letters are significantly differed at P < 0.05.