An 8-week feeding experiment was carried out to explore the effects of dietary n-3/n-6 polyunsaturated fatty acid (PUFA) ratio on growth performance, lipid metabolism, hepatic antioxidant status, and gut flora of spotted seabass (Lateolabrax maculatus). Six experimental diets were formulated to contain different levels of two purified oil sources including docosahexaenoic and eicosapentaenoic acids enriched oil (n-3) and linoleic acid-enriched oil (n-6) leading to n-3/n-6 PUFA ratios of 0.04, 0.35, 0.66, 1.35, 2.45 and 16.17. Each diet was fed to triplicate groups of juvenile L.maculatus (11.06 ± 0.20g, 30 fish/tank). Final body weight (FBW), weight gain (WG), specific growth rates (SGR), protein efficiency ratio (PER) and feed utilization efficiency increased as n-3/n-6 PUFA ratio increased up to a certain level, and then decreased thereafter. Fish fed the diet with n-3/n-6 PUFA ratio of 0.66 exhibited the highest FBW, WG, SGR and PER and the lowest feed conversion ratio. Lower n-3/n-6 PUFA ratios induced up-regulated expression of lipid synthesis-related genes (fas,acc2 and srebp-1c) and down-regulated expression of lipolysis related genes (atgl, pparα, cpt-1 and aox). Higher expression of lipolysis-related genes (atgl, pparα and cpt-1) was recorded at moderate n-3/n-6 PUFA ratios (0.66 to 1.35). Moreover, inappropriate n-3/n-6 PUFA ratios triggered up-regulation of pro-inflammatory genes (il-6 and tnf-α) and down-regulation of anti-inflammatory genes (il-4 and il-10) in the intestine. The diet with n-3/n-6 PUFA ratio of 0.66 inhibited intestine inflammation, improved intestinal flora richness, increased the abundance of beneficial bacteria such as Lactobacillus, Alloprevotella and Ruminococcus, and reduced the abundance of harmful bacteria including Escherichia-Shigella and Enterococcus. In summary, it could be suggested that a dietary n-3/n-6 PUFA ratio of 0.66 can improve growth performance and feed utilization in L.maculatus, as is deemed to be mediated through regulation of lipid metabolism and intestinal flora.

Fish gut barrier damage under intensive culture model is a significant concern for aquaculture industry. This study aimed to investigate the effects of bile acids (BAs) on gut barriers in Micropterus salmoides. A germ-free (GF) zebrafish model was employed to elucidate the effects of the direct stimulation of BAs and the indirect regulations mediated by the gut microbiota on gut barrier functions. Four diets were formulated with BAs supplemented at 0, 150, 300 and 450 mg/kg, and these 4 diets were defined as control, BA150, BA300 and BA450, respectively. After 5 weeks of feeding experiment, the survival rate of fish fed with BA300 diet was increased (P < 0.05). Histological analysis revealed an improvement of gut structural integrity in the BA150 and BA300 groups. Compared with the control group, the expression of genes related to chemical barrier (mucin, lysozyme and complement 1) and physical barrier (occludin and claudin-4) was increased in the BA150 and BA300 groups (P < 0.05), and the expression of genes related to immunological barrier (interleukin [IL]-6, tumor growth factor β,IL-10, macrophage galactosetype lectin and immunoglobulin M [IgM]) was significantly increased in the BA300 group (P< 0.05), but the expression of genes related to chemical barrier (hepcidin) and immunological barrier (IL-1β, tumor necrosis factor-α, IL-6 and arginase) was significantly decreased in the BA450 group (P< 0.05). Gut microbiota composition analysis revealed that the abundance of Firmicutes was augmented prominently in the BA150 and BA300 groups (P< 0.05), while that of Actinobacteriota and Proteobacteria showed a downward trend in the BA150 and BA300 groups (P> 0.05). The results of the gut microbiota transferring experiment demonstrated an upregulation of gut barrier-related genes, including immunoglobulin Z/T (IgZ/T), IL-6, IL-1β and IL-10, by the gut microbiota transferred from the BA300 group compared with the control (P< 0.05). Feeding the BA300 diet directly to GF zebrafish resulted in enhanced expression of IgM, IgZ/T, lysozyme, occludin-2, IL-6 and IL-10 (P< 0.05). In conclusion, BAs can improve the gut barriers of fish through both direct and indirect effects mediated by the gut microbiota.

Antibiotic resistance of pathogens, which is caused by the abuse of in-feed antibiotics, threatens the sustainable development of livestock production. The present study aimed to investigate the efficiency of porcine intestinal antimicrobial peptide (PIAP) as an alternative to in-feed antibiotics in terms of growth performance, intestinal morphology, digestive enzymes and immunity, and microbiota community of the post-weaning piglets. A total of 204 piglets (Duroc × Landrace × Yorkshire, weaned at 28 d age) with a similar body weight of 7.97 ± 1.04 kg were randomly allocated to 4 groups (51 piglets per group): (1) control group: basal diet; (2) AB group: antibiotic, basal diet þ chlortetracycline (1000mg/kg from d 1 to 24; 500mg/kg from d 25 to 37); (3) P1 group: basal diet þ a relatively low dose of PIAP (400mg/kg from d 1 to 24; 300mg/kg from d 25 to 37); (4) P2 group, basal diet þ a relatively high dose of PIAP (600mg/kg from d 1 to 24; 500mg/kg from d 25 to 37). The results showed that serum indicators of hepatocyte damage and relative organ weight were not affected by these treatments (P > 0.05). Compared with the AB treatment, the P1 treatment remarkably decreased jejunal crypt depth and increased jejunal and ileal villus height:crypt depth ratio (P < 0.05). The values of jejunal maltase, lactase, sucrase, intestinal alkaline phosphatase, and secretory immunoglobulin A (SIgA) in the P1 group were sharply increased compared with those in the control and P2 groups (P < 0.05). Compared with the control group, the P1 group decreased serum concentrations of D-lactate, diamine oxidase, and endotoxin (P < 0.05), and increased the abundance of Lactobacillus reuteri (P < 0.05) in the colonic feces. Furthermore, there was a positive correlation between the abundance of L. reuteri and the concentrations of maltase, lactase, sucrase, and SIgA (P < 0.05). Collectively, dietary supplementation with a relatively low dose of PIAP (400 mg/kg from d 1 to 24; 300 mg/kg from d 25 to 37) demonstrates beneficial effects on intestinal morphology, digestive enzymes, immunity, and permeability by shaping the gut microbiota composition in weaned piglets. This study will provide a valuable reference for using PIAP as an in-feed antibiotic alternative in swine production.

Carbohydrates have a protein sparing effect, but long-term feeding of a high-carbohydrate diet (HCD) leads to metabolic disorders due to the limited utilization efficiency of carbohydrates in fish. How to mitigate the negative effects induced by HCD is crucial for the rapid development of aquaculture. Uridine is a pyrimidine nucleoside that plays a vital role in regulating lipid and glucose metabolism, but whether uridine can alleviate metabolic syndromes induced by HCD remains unknown. In this study, a total of 480 Nile tilapia (Oreochromis niloticus) (average initial weight 5.02 ± 0.03g) were fed with 4 diets, including a control diet (CON), HCD, HCD + 500 mg/kg uridine (HCUL) and HCD + 5,000 mg/kg uridine (HCUH), for 8 weeks. The results showed that addition of uridine decreased hepatic lipid, serum glucose, triglyceride and cholesterol (P < 0.05). Further analysis indicated that higher concentration of uridine activated the sirtuin1 (sirt1)/adenosine 5-monophosphate-activated protein kinase (AMPK) signaling pathway to increase lipid catabolism and glycolysis while decreasing lipogenesis (P < 0.05). Besides, uridine increased the activity of glycogen synthesis-related enzymes (P < 0.05). This study suggested that uridine could alleviate HCD-induced metabolic syndrome by activating the sirt1/AMPK signaling pathway and promoting glycogen synthesis. This finding reveals the function of uridine in fish metabolism and facilitates the development of new additives in aquatic feeds.

Rabbit breeding has many critical aspects related to reproduction, production, and animal welfare, which reduce its profitability as well as consumer attractiveness. Dietary supplementation with n-3 polyunsaturated fatty acids (PUFA) seems to be a good nutritional strategy to improve several aspects of rabbit breeding, enhance animal welfare and produce a new functional food considered healthy for human consumption. For this reason, the main available scientific research regarding the physiological effects of n-3 PUFA rich products supplemented to the rabbit diet will be reviewed. In particular, consequences on the reproductive performances of both doe and buck, the productive parameters, and the meat quality will be analysed.

Alfalfa (Medicago sativa L.) is a legume forage that is widely cultivated owing to its high biomass yield and favorable nutrient values. However, alfalfa contains relatively high lignin, which limits its utilization. Downregulation of two transcriptional factors, Transparent Testa8 (TT8) and Homeobox12 (HB12), has been proposed to reduce lignin content in alfalfa. Therefore, silencing of TT8 (TT8i) and HB12 (HB12i) in alfalfa was achieved by RNAi technology. The objective of this project was to determine effect of gene modification through silencing of TT8 and HB12 genes in alfalfa plants on lignin and phenolic content, bioenergic value, nutrient supply from rumen degradable and undegradable fractions, and in vitro ammonia production in response to the silencing of TT8 and HB12 genes in alfalfa. All gene silenced alfalfa plants (5 TT8i and 11 HB12i) were grown under greenhouse conditions with wild type as a control. Samples were analyzed for bioactive compounds, degradation fractions, truly digestible nutrients, energetic values and in vitro ammonia productions in ruminant systems. Furthermore, relationships between physiochemical, metabolic and fermentation characteristics and molecular spectral parameters were determined using vibrational molecular spectroscopy. Results showed that the HB12i had higher lignin, while TT8i had higher phenolics. Both silenced genotypes had higher rumen slowly degraded carbohydrate fractions and truly digestible neutral detergent fiber, but lower rumen degradable protein fractions. Moreover, the HB12i had lower truly digestible crude protein, energetic values and ammonia production compared with other silenced genotypes. In addition, in relation to the nutritive values of alfalfa, structural carbohydrate parameters were negatively correlated, whereas alpha/beta ratio in protein structure was positively correlated. Furthermore, good predictions were obtained for degradation of protein and carbohydrate fractions and energy values from molecular spectral parameters. In conclusion, silencing of the TT8 and HB12 genes decreased protein availability and increased fiber availability. Silencing of the HB12 gene also increased lignin and decreased energy and rumen ammonia production. Moreover, nutritional alterations were closely correlated with molecular spectral parameters. Therefore, gene modification through silencing the TT8 and HB12 genes in alfalfa influenced physiochemical, metabolic and fermentation characteristics.

Butyrate promotes the growth and gastrointestinal development of calves. But, the mechanisms behind its effects on signaling pathways of the gastrointestinal tract and rumen microbiome is unclear. This study aimed to reveal transcriptomic pathways of gastrointestinal epithelium and microbial community in response to butyrate supplementation in calves fed a high fiber starter. Fourteen Holstein bull calves (39.9 ± 3.7 kg, 14 d of age) were assigned to 2 groups (sodium butyrate group, SB; control group, Ctrl). The SB group received 0.5% SB supplementation. At d 51, the calves were slaughtered to obtain samples for analysis of the transcriptome of the rumen and jejunum epithelium as well as ruminal microbial metagenome. Sodium butyrate supplementation resulted in a higher performance in average daily gain and development of jejunum and rumen papillae. In both the rumen and jejunum epithelium, SB down-regulated pathways related to inflammation including NF-κB (PPKCB, CXCL8, CXCL12), interleukin-17 (IL17A, IL17B, MMP9), and chemokine (CXCL12, CCL4, CCL8) and up-regulated immune pathways including the intestinal immune network for immunoglobulin A (IgA) production (CD28). Meanwhile, in the jejunum epithelium, SB regulated pathways related to nutritional metabolism including nitrogen metabolism (CA1, CA2, CA3), synthesis and degradation of ketone bodies (HMGCS2, BDH1, LOC100295719), fat digestion and absorption (PLA2G2F, APOA1, APOA4), and the PPAR signaling pathway (FABP4, FABP6, CYP4A11). The metagenome showed that SB greatly increased the relative abundance of Bacillus subtilis and Eubacterium limosum, activated ruminal microbial carbohydrate metabolism pathways and increased the abundance of carbohydrate hydrolysis enzymes. In conclusion, butyrate exhibited promoting effects on growth and gastrointestinal development by inhibiting inflammation, enhancing immunity and energy harvesting, and activating microbial carbohydrate metabolism. These findings provide new insights into the potential mechanisms behind the beneficial effects of butyrate in calf nutrition.

This experiment aimed to study the effects of supplemental methionine sources, 2-hydroxy-4 methyl(thio) butanoic acid (HMTBa) and DL-Methionine (DL-Met), on productive performance, egg quality, and redox status of laying ducks. A total of 792 healthy 25-wk-old Longyan laying ducks with similar body weights were randomly allotted to 11 treatment groups. Each treatment group had 6 replicates of 12 ducks. The trial lasted for 16 wk. Ducks were fed a basal deficient diet (Met: 0.24%; Met + Cys: 0.51%) or supplemented with DL-Met or HMTBa at 0.05%, 0.12%, 0.19%, 0.26%, and 0.33% of diet, respectively. Compared with the basal diet, supplementation with either DL-Met or HMTBa increased the average egg weight, egg mass, and decreased feed to egg ratio during the whole trial period (P < 0.05). Albumen weight and its ratio to total egg weight were increased, but yolk and shell ratio, albumen height, Haugh unit and shell breaking strength were decreased (P < 0.05). Dietary DL-Met or HMTBa supplementation increased taurine, methionine, leucine, tryptophan and arginine content, and decreased serine and lysine content in plasma (P < 0.05). The redox status of laying ducks was improved by enhancing the glutathione peroxidase and catalase activities, glutathione content and its ratio relative to glutathione (oxidized) content and decreasing malondialdehyde content and increasing mRNA expression of superoxide dismutase-1, glutathione peroxidase-1, hemeoxygenase-1 and nuclear factor-like 2 in liver and ileum with the supplementation of DL-Met or HMTBa (P < 0.05). Liver health status measured by average area proportion lipid droplet was improved with supplementation of DL-Met or HMTBa (P < 0.05). Villus height and villus height to crypt depth ratio in the ileum and the ileal gene expression of tight junction protein and occludin were increased with DL-Met or HMTBa supplementation (P < 0.05). Taken together, these results suggested that the efficacy of dietary supplementation of HMTBa was similar to DL-Met, and it ranged from 98% to 100% for productive performance and egg albumen ratio in laying ducks (25 to 41 wk).

This study explored the variation of ileal endogenous amino acid (IEAA) losses and its influencing factors in chickens offered nitrogen-free diets (NFD) containing different ratios of amylose to amylopectin (AM/AP). A total of 252 broiler chickens at 28d old were randomly allocated into 7 treatment groups for a 3d trial. The dietary treatments included a basal diet (control), a NFD containing corn starch (CS), and 5 NFD with AM/AP ratios of 0.20, 0.40, 0.60, 0.80, and 1.00, respectively. As the AM/AP ratio increased, the IEAA losses of all AAs, starch digestibility and maltase activity linearly decreased (P < 0.05), but the DM digestibility linearly and quadratically decreased (P < 0.05). Compared with the control, the NFD increased the number of goblet cells and its regulatory genes mucin-2 and krüppel-like factor 4 (KLF-4) while decreasing serum glucagon and thyroxine concentrations, ileal villus height, and crypt depth (P < 0.05). Additionally, NFD with lower AM/AP ratios (0.20 and 0.40) decreased the ileal microbiota species richness (P < 0.05). In all NFD groups, the number of Proteobacteria increased whereas the abundance of Firmicutes dropped (P < 0.05). However, the broilers in the AM/AP 0.60 group were closer to the digestive physiological state of chickens fed the control diet, with no significant change in maltase activity and mucin-2 expression (P < 0.05). In conclusion, increasing AM/AP ratio in a NFD decreased the IEAA losses and the apparent ileal digestibility of starch but inevitably resulted in malnutrition and disruption of gut microbiota homeostasis. This study recommends AM/AP in NFD at 0.60 to measure IEAA of broiler chickens.