Integrated analysis of microbiome and host transcriptome reveals the damage/protective mechanism of corn oil and olive oil on the gut health of grouper (♀ Epinephelus fuscoguttatus × â™‚ E. lanceolatu).

As digestive and immune organs of animals, the gut was frequently used to evaluate the health status of aquatic animals. In previous oil source alternatives study, corn oil (CO) had been found to induce gut inflammation, while olive oil (OO) had been found to be effective in protecting intestinal health. Three diets with different oil sources (fish oil, CO, OO) were formulated for an 8-week culture experiment, and it was proposed to combine 16S sequencing and transcriptome sequencing analysis to preliminarily elucidate the damage/protection mechanism of CO and OO on the gut health of grouper (♀ Epinephelus fuscoguttatus × â™‚ E. lanceolatu). We found that CO indeed damaged to gut health and destroyed the gut structure, while OO had a positive outcome in protecting the gut structure, promoting digestibility and relieving enteritis. Photobacterium, Romboutsia and Epulopiscium were significantly enriched in OO group and Staphylococcus were significantly enriched in CO group. Transcriptome sequencing further revealed CO could activated Complement and coagulation cascades, Staphylococcus aureus infection, Systemic lupus erythematosus, and Tuberculosis pathways; conversely, OO activated B-cell signaling receptors, promoted B-cell proliferation and apoptosis, and thus activated B-cell signaling pathways to enhance immunity, whereas OO can regulate IL17 signaling pathway and TNF signaling pathway to inhibit NF-κB signaling pathway to reduce pro-inflammatory response. By integrating the microbiome and transcriptome, further identified all differential microorganisms were directly and significantly correlated with differential genes, and Clostridium_sensu_stricto_1, Romboutsia, Staphylococcus might as the core regulates the expression of differential gene in the organism. These results reveal that different oil sources alter gut gene expression mainly by modulating the composition and abundance of gut microbiota, further regulating the health status of the gut. Gut microbiota could be used as biomarkers to provide reference and solutions for the mitigation of inflammation in aquatic animals.

Effects of choline supplementation on growth performance, liver histology, nonspecific immunity and related genes expression of hybrid grouper (♀ Epinephelus fuscoguttatus × â™‚ E. lanceolatu) fed with high-lipid diets.

This study was conducted to evaluate the effect of dietary choline levels on growth performance, liver histology, nonspecific immunity and related gene expression of hybrid grouper (♀ Epinephelus fuscoguttatus × â™‚ E. lanceolatus) fed with high-lipid diets. The fish (initial body weight 6.86 ± 0.01 g) were fed diets containing different choline levels (0, 5, 10, 15, and 20 g/kg, named D1, D2, D3, D4, and D5, respectively) for 8 weeks. The results showed that:(1) dietary choline levels had no significant effect on final body weight (FBW), feed conversion rate (FCR), visceral somatic index(VSI) and condition factor (CF) compared with the control group (P > 0.05). However, the hepato somatic index (HSI) in the D2 group was significantly lower than that in the control group and the survival rate (SR) in the D5 group was significantly lower (P < 0.05). (2) with dietary choline level increasing, alkaline phosphatase (AKP) and superoxide dismutase (SOD) of serum showed a tendency to increase and then decrease, and the maximum values were obtained in the D3 group, but the contents of alanine aminotransferase (ALT) and aspartate aminotransferase (AST) decreased significantly (P < 0.05). (3) Immunoglobulin M (IgM), lysozyme (LYZ), catalase (CAT), total antioxidative capacity (T-AOC), and SOD in the liver all showed a trend of first increase and then decrease with the dietary choline level increased, and all of them achieved the maximum value at D4 group (P < 0.05), while reactive oxygen species (ROS) and malondialdehyde (MDA) in the liver decreased significantly (P < 0.05). (4) results from liver sections suggest that appropriate levels of choline can improve cell structure, compared with the control group, the damaged histological morphology of the liver was relieved and even returned to normal in D3 group. (5) in the D3 group, choline significantly upregulated the expression of hepatic sod and cat mRNA, whereas the expression of cat in the D5 group was significantly lower than that in the control group (P < 0.05); And the supply of choline stimulated a significant down-regulation of interleukin 6 (il6), myeloid differentiation factor 8 (myd88), toll-like receptor 22 (tlr22) mRNA expression levels in liver, while the expression of cellular tumor antigen p53 (p53) and interleukin 10 (il10) showed an upward and then downward trend (P < 0.05). In general, choline can improve the immunity of hybrid grouper by regulating non-specific immune-related enzyme activity and gene expression and reducing oxidative stress induced by high-lipid diet.

Substituting fish meal with castor meal in diets of hybrid grouper (Epinephelus fuscoguttatus♀ × E. lanceolatus♂): Effects on growth performance, immune response, antioxidant and digestive enzyme activities, gut morphology, and inflammatory-related gene expression.

The present study was conducted to investigate the effects of replacing fishmeal (FM) with castormeal (CM) on the growth performance, immune response, antioxidant and digestive enzyme activities, intestinal morphology, and expression of inflammatory-related genes in juvenile hybrid grouper (Epinephelus fuscoguttatus♀ ×E. lanceolatus♂). Six iso-nitrogenous (50% crude protein) and iso-lipidic (10% crude lipid) diets were formulated; namely, a reference diet (FM) containing 50% FM and five experimental diets (4% (CM4), 8% (CM8), 12% (CM12), 16% (CM16), and 20% (CM20)) in which FM protein was substituted with CM at varying levels to feed fish (initial weight: 9.12 ± 0.01 g) for 8 weeks. The results showed that the final weight, weight gain rate, and specific growth rate were highest in the FM, CM4, and CM8 groups, whereas the feed conversion ratio, hepatosomatic and viscerosomatic indexes were significantly enhanced in the CM4 group in comparison to the others. The CM4 and CM12 groups were observed to show the highest intestinal length index values compared to the other groups, with the CM20 revealing the worst growth performance. The serum total protein content first increased (P < 0.05) in the CM4 group and decreased (P < 0.05) afterward. Nonetheless, a decreasing significant (P < 0.05) cholesterol and triglyceride contents were witnessed with the increasing replacement of FM with CM. Compared to the control group, a significant increase (P < 0.05) in the activities of serum and liver immunoglobulin-M, superoxide dismutase, glutathione peroxidase, total antioxidant capacity, and complement-3 (except serum activity for CM12 group); liver lysozyme; intestinal amylase, and lipase, was witnessed in the CM groups. However, the serum lysozyme activity was highest (P < 0.05) in the CM4 group and lowest in the CM20 group. While the least serum malondialdehyde contents were observed in the CM4 group, that of the liver malondialdehyde was least witnessed in the FM, CM4, CM8, CM12, and CM16 groups as compared to the CM20. The intestinal histological examination revealed a significantly decreasing trend for villi height and villi width with increasing replacement levels. However, the muscle thickness, crypt depth, and type II mucus cells first increased upto 4% replacement level and later decreased. The increasing of dietary replacement levels significantly up-regulated pro-inflammatory (il-1ß, tnf-α, myd88, ifn-γ, tlr-22, and il-12p40) and down-regulated anti-inflammatory (il-10, tgf-ß, mhc-iiß) and anti-bacterial peptide (epinecidin and hepcidin) mRNA levels in the intestine. The mRNA levels of il-6 was up-regulated firstly upto 4 and 8% replacement levels, and later down-regulated with increasing replacement. These results suggested that, although higher dietary CM replacement enhances the immune, antioxidant and digestive enzymes, it aggravates intestinal inflammation. Replacing 4 and 8% of FM with CM could enhance the growth performance of fish.

Impacts of tea polyphenols on growth, antioxidant capacity and immunity in juvenile hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂) fed high-lipid diets.

The experiment aimed to investigate the alteration of tea polyphenols (TP) in growth and immunity for hybrid grouper (Epinephelus fuscoguttatus ♀ × E. lanceolatus ♂) fed high-lipid diets. Six concentrations of TP (0, 0.01, 0.02, 0.04, 0.08, 0.16%, named TP1 (basic diet control), TP2, TP3, TP4, TP5, TP6) were supplied in isonitrogenous (51%) and isolipidic (16.7%) experimental diets. These diets were fed to the juvenile grouper (8.68 ± 0.22 g) for 8 weeks. The results showed that dietary TP significantly increased the weight gain rate and specific growth rate (P < 0.05), compared with the control group. The protein efficiency ratio in TP4 group was significantly higher than that of the control group (P < 0.05). TP supplement in high-lipid diets increased antioxidant capacity in the serum (CAT, GSH-Px, T-AOC) and liver (SOD, CAT, GSH-Px, T-AOC). Additionally, dietary TP decreased oxidative stress (ROS, MDA) and improved immunity (ACP, AKP, LYS, IgM) in the liver. The histology of hepatic tissue indicated that dietary TP alleviated pathological symptoms caused by high-lipid diets. Compared with the control group, appropriate dietary TP significantly up-regulated expression of sod, cat, gsh-px, nrf2, keap1, hsp70, hsp90, myd88, tnfα and down-regulated expression of tlr22, il8, il1ß, il10 in the liver (P < 0.05). In the head kidney, expression of myd88, il1ß, tnfα and il6 were significantly up-regulated and expression of tlr22 and il10 were significantly down-regulated by dietary TP (P < 0.05). After the challenge of Vibrio harveyi, survival rate in higher doses of TP group (TP4 âˆ¼ TP6) was evidently higher, compared with the control group. In conclusion, TP supplement in high-lipid diets improved antioxidant capacity and enhanced immunity of grouper. We speculate that TP may play the role of an immunostimulant, enhancing immunity and disease resistance by cytokine-medicated immune responses. Based on the second-order regression, 0.092-0.106% tea polyphenols were recommended in juvenile grouper high-lipid diets.

The positive effects of dietary inositol on juvenile hybrid grouper (♀ Epinephelus fuscoguttatus × â™‚ E. lanceolatu) fed high-lipid diets: Growthperformance, antioxidant capacity and immunity.

The objective of the present research was to assess the influence of inositol supplementation on growth performance, histological morphology of liver, immunity and expression of immune-related genes in juvenile hybrid grouper (♀ Epinephelus fuscoguttatus × â™‚ E. lanceolatu). Hybrid grouper (initial weight 6.76 ± 0.34 g) were fed isonitrogenous and isolipidic diets (16%) with various inositol levels of 0.17 g/kg (J1, the control group), 0.62 g/kg (J2), 1.03 g/kg (J3), 1.78 g/kg (J4), 3.43 g/kg (J5), 6.59 g/kg (J6), respectively. The growth experiment lasted for 8 weeks. The results indicated that dietary inositol had a significant promoting effect on final mean body weight of the J5 and J6 groups and specific growth rate (SGR) of the J3, J4, J5 and J6 groups (P < 0.05). In the serum, superoxide dismutase (SOD) of the J4 group became significantly active compared with that of the control group (P < 0.05), while aspartate transaminase (AST), alanine transaminase (ALT) and alkaline phosphatase (AKP) activities in the inositol-treated groups showed distinctly decreased compared with those of the control group (P < 0.05). In the liver, dietary inositol could significantly increase the activities of SOD, catalase (CAT), lysozyme (LYZ) and the contents of total antioxidative capacity (T-AOC) and immunoglobulin M (IgM) (P < 0.05), and distinctly reduce the content of malondialdehyde (MDA) as well as reactive oxygen species (ROS) (P < 0.05). Compared with the control group, the damaged histological morphology of the liver was relieved and even returned to normal after an inositol increase (0.4-3.2 g/kg). In the liver, the remarkable up-regulation of SOD, CAT, glutathione peroxidase (GPX), heat shock protein70 (HSP70) and heat shock protein90 (HSP90) expression levels were stimulated by supply of inositol, while interleukin 6 (IL6), interleukin 8 (IL8) and transforming growth factor ß (TGF-ß) expression levels were down-regulated by supply of inositol. In head kidney, the mRNA of toll-like receptor 22 (TLR22), myeloid differentiation factor 88 (MyD88) and interleukin 1ß (IL1ß) expression levels were significantly down-regulated (P < 0.05), which could further lead to remarkable down-regulation of IL6 and tumor necrosis factor α (TNF-α) expression (P < 0.05). These results indicated that high-lipid diets with supply of inositol promoted growth, increased the antioxidant capacity, and suppressed the inflammation of the liver and head kidney by inhibiting the expression of pro-inflammation factors (IL6, IL8, TGF-ß and TNF-α). In conclusion, these results indicated that dietary inositol promoted growth, improved antioxidant capacity and immunity of hybrid grouper fed high-lipid diets. Based on SGR, broken-line regression analysis showed that 1.66 g/kg inositol supply was recommended in high-lipid diets of juvenile grouper.