Normally, proper fermentation can be an efficient and widely used method to improve feed quality in animal rearing; however, the studies on crustaceans, especially Eriocheir sinensis, remain limited. This study aimed to investigate whether feed fermentation could meliorate dietary nutritional value and benefit E. sinensis rearing. First, non-fermented feed (NFD) and fermented feed (FD) were produced and assessed, respectively. Then, the "Y" maze feed choice behavior test (180 times; 30 times, 6 rounds) was conducted to assess the attractiveness of these 2 feeds for crabs. Finally, a total of 80 crabs (44.10 ± 0.80 g) were randomly assigned into 2 groups with 4 replicates, and fed the experimental diets for 8 weeks to evaluate the effects of each feed on growth, antioxidant capacity, meat flavor, and intestinal microbiota. In this study, FD showed higher levels of crude protein (P < 0.01), soluble protein (P < 0.01), amino acids (P < 0.05), lactic acid (P < 0.001), and lower levels of crude fiber (P < 0.05) and antinutritional factors (agglutinin, trypsin inhibitor, glycinin, and β-conglycinin) (P < 0.001) than NFD. Additionally, FD was more attractive to crabs than NFD (P < 0.01) and it stimulated the appetite of crabs more than NFD (P < 0.05). The growth performance, feed efficiency, and digestive enzyme activity of FD-fed crabs were significantly higher than those of NFD-fed crabs (P < 0.05). The electronic sensory measurements and free amino acid profiles revealed that the FD diet had positive impacts on the meat flavor of crabs, particularly in "sweet" and "umami" tastes. Moreover, the antioxidant capacity of FD-fed crabs was significantly higher than that of NFD-fed crabs (P < 0.05). Fermented feed also affected the diversity and composition of intestinal microflora. The functional prediction of microbial communities showed that crabs fed FD had a better microecological environment in the intestine. In conclusion, the fermentation of aquafeed could be an effective approach to enhance feed quality and therefore benefit E. sinensis rearing.

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).

Oxidative stress is a common phenomenon in poultry production. Several molecules, including antioxidant genes, miRNAs, and gut microbiota metabolites, have been reported to participate in redox regulation. Lactiplantibacillus plantarum P8 (P8) was shown to improve the antioxidant capacity of chickens, but the specific molecular mechanisms remain unclear. In this study, 400 broilers were allocated to 4 treatment groups: control diet (Con group), control diet ＋ dexamethasone injection (DEX group), control diet containing 1 × 10⁸ CFU/g P8 (P8 group), and control diet containing 1 × 10⁸ CFU/g P8 + DEX injection (DEX_P8 group). Integrated analysis of the microbiome, metabolomics, and miRNAomics was conducted to investigate the roles of P8 in oxidative stress in broilers. Results demonstrated that P8 supplementation significantly improved growth performance, jejunal morphology, and antioxidant function in DEX-treated broilers. Analysis of the gut microbiota revealed a higher abundance of Barnesiella (P = 0.01) and Erysipelatoclostridium (P = 0.05) in the DEX_P8 group than in the DEX group. Functional prediction indicated that certain pathways, including the phenylacetate degradation pathway, were enriched in the DEX_P8 group compared to the DEX group. Metabolites in the cecal contents were distinct between the groups. P8 supplementation increased the content of metabolites with antioxidant capacity, e.g., urobilinogen (P < 0.01), and decreased that of metabolites related to oxidative stress, e.g., genistein (P < 0.01). Functional prediction indicated that metabolites that differed between the DEX_P8 and DEX groups were enriched in pathways including “tryptophan metabolism” and “primary bile acid biosynthesis”. The miRNAomics analysis further showed that, compared to the DEX group, several miRNAs in the jejunum, such as gga-miR-21-3p (P = 0.03), were increased, whereas gga-miR-455-3p (P = 0.02) was decreased in the DEX_P8 group. The PI3K-Akt, Ras, and Rap1 signaling pathways were enriched in the DEX_P8 group compared to the DEX group through KEGG analysis. Correlation analysis revealed potential interactions between growth performance, oxidation/antioxidation, jejunal morphology, gut microbiota, cecal content metabolites, and jejunal miRNAs. Overall, our results indicate that P8 supplementation may improve the growth performance, jejunal morphology and antioxidant capacity of DEX-treated broilers by regulating gut microbiota, its metabolites, and intestinal miRNAs.

The alterations in feed ingredients and the nutrient matrix to produce reduced-protein diets may affect bone morphology and mineralization in laying hens. This study was implemented to determine the effects of L-arginine (Arg), guanidinoacetic acid (GAA), and L-citrulline (Cit) supplementation to Arg-deficient reduced-protein diets on bone morphology, strength, and mineralization status of laying hens. Individually housed Hy-Line Brown laying hens were evenly distributed to five dietary treatments with 25 replicates per treatment from 20 to 40 wk of age. Treatments consisted of a standard protein diet (17% crude protein, SP), a reduced-protein diet deficient in Arg (13% crude protein, RP), and RP supplemented with Arg (0.35% Arg, RP-Arg), GAA (0.46% GAA equivalent to 0.35% Arg, RP-GAA), or Cit (0.35% Cit equivalent to 0.35% Arg, RP-Cit) to reach the Arg level of SP diets. Birds fed the SP diet had similar bone weight, ash, length, width, Seedor index, breaking strength, and serum mineral concentration, but higher toe B level (P < 0.001) compared to those fed the RP diet at wk 40. Birds fed the SP diet consumed more but also excreted more K and B compared to those fed the RP diet (P < 0.01). Birds fed the SP diet had lower Cu digestibility (P = 0.01) and higher B retention (P < 0.01) compared to those offered the RP diet. Supplementation of Arg, GAA, and Cit to the RP diet increased relative femur weight and length (P < 0.001). Citrulline supplementation also increased relative tibia and femur ash, and Zn digestibility (P < 0.05). Supplementation of GAA to the RP diet decreased serum Ca, P, and Mg levels, decreased tibia Fe and Mg levels and toe Mg level, but increased Al, Fe, Zn, and Mn digestibility (P < 0.05). The current findings demonstrated the capacity of laying hens to adapt to low mineral intake by increasing mineral utilization. Overall, bone morphology and breaking strength, and serum mineral level in laying hens were not influenced by dietary CP levels. Dietary Arg, GAA, or Cit supplementation were effective in improving bone morphology and mineralization in laying hens fed Arg-deficient RP diets.

Several reports have revealed the vital role that probiotics play in fish growth and health. However, few works are available for host gut-derived probiotics on the growth, immunity, and gut microbiota of fish, especially in hybrid grouper (♀Epinephelus fuscoguttatus × ♂Epinephelus lanceolatus) due to their isolation difficulty and functional verification. This study aimed at assessing 3 host gut-derived Bacillus species' effects on the growth, immune and antioxidant-biochemical responses, haematological parameters, intestinal morphology, immune-related gene expression, gut microbiota, and disease resistance against Vibrio harveyi in hybrid grouper. A total of 480 hybrid grouper (initial weight = 9.03 ± 0.02 g) were randomly allotted into 4 groups, namely, the group fed a basal diet without probiotic inclusion (control, B0), the group fed the basal diet with Bacillus velezensis GPSAK4 (BV), the group fed the basal diet with Bacillus subtilis GPSAK9 (BS), and the group fed the basal diet with Bacillus tequilensis GPSAK2 (BT) strains at 1.0 × 10⁹ CFU/g. After a 6-week feeding trial, the results revealed significant improvements (P < 0.05) in the growth performance, whole fish-body proximate composition, blood haematological parameters, serum, liver, and intestinal biochemical indexes, intestinal morphology, and protection against V. harveyi pathogen in the probiotic-treated groups compared with the untreated. Additionally, the expressions of intestinal tight junction genes (occludin and ZO1), pro- and anti-inflammatory genes, including IL1β, IL6, IL8, TNFα, MyD88, IL10, and TGFβ, were upregulated (P < 0.05) after Bacillus species administration. Host gut-derived Bacillus supplementation shaped the gut microbiota by significantly increasing (P < 0.05) the relative abundance of Proteobacteria, Bacteroidetes, Actinobacteria (except the BS group), Acidobacteria (except the BT group), Cyanobacteria (except the BV and BT groups), and Verrucomicrobia phyla, as well as known beneficial genera (Romboutsia, Turicibacter, Epulopiscium, Clostridium_sensu_stricto 1 and 13, Lactobacillus, and Bacillus), but significantly decreased (P < 0.05) the abundance of Firmicutes, Chloroflexi, and Fusobacteria phyla, and purported pathogenic genera (Staphylococcus and Photobacterium) compared with the control group. Collectively, the results suggest that B. velezensis GPSAK4, B. subtilis GPSAK9 (especially this strain), B. tequilensis GPSAK2 dietary supplementation at 1.0 × 10⁹ CFU/g has positive effects on the intestinal health of hybrid grouper via microbial composition modulation, thus enhancing the assimilation and absorption of nutrients to boost fish growth, immunity, and disease resistance.

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.

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.

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.

Lysozyme (LZ) is a purely natural, nonpolluting and nonspecific immune factor, which has beneficial effects on the healthy development of animals. In this study, the influences of LZ on the growth performance and intestinal barrier of weaned piglets were studied. A total of 48 weaned piglets (Landrace × Yorkshire, 22 d old) were randomly divided into a control group (basal diet) and a LZ group (0.1% LZ diet) for 19 d. The results showed that LZ could significantly improve the average daily gain (ADG, P < 0.05) and average daily feed intake (ADFI, P < 0.05). LZ also improved the intestinal morphology and significantly increased the expression of occludin in the jejunum (P < 0.05). In addition, LZ down-regulated the expression of interleukin-1β (IL-1β, P < 0.05) and tumor necrosis factor-a (TNF-α, P < 0.05), and inhibited the expression of the genes in the nuclear factor-k-gene binding (NF-κB, P<0.05) signaling pathway. More importantly, the analysis of intestinal flora showed LZ increased the abundance of Firmicutes (P < 0.05) and the ratio of Firmicutes to Bacteroidota (P = 0.09) at the phylum level, and increased the abundance of Clostridium_sensu_stricto_1 (P < 0.05) and reduced the abundance of Olsenella and Prevotella (P < 0.05) at the genus level. In short, this study proved that LZ could effectively improve the growth performance, relieve inflammation and improve the intestinal barrier function of weaned piglets. These findings provided an important theoretical basis for the application of LZ in pig production.

Vitamin A and its metabolite, retinoic acid (RA) play important roles in regulating skeletal muscle development. This study was conducted to investigate the effects of early intramuscular vitamin A injection on the muscle growth of lambs. A total of 16 newborn lambs were given weekly intramuscular injections of corn oil (control group, n = 8) or 7,500 IU vitamin A palmitate (vitamin A group, n = 8) from birth to 3 wk of age (4 shots in total). At 3 wk of age and weaning, biceps femoris muscle samples were taken to analyze the effects of vitamin A on the myogenic capacity of skeletal muscle cells. All lambs were slaughtered at 8 months of age. The results suggest that vitamin A treatment accelerated the growth rate of lambs and increased the loin eye area (P < 0.05). Consistently, vitamin A increased the diameter of myofibers in longissimus thoracis muscle (P < 0.01) and increased the final body weight of lambs (P < 0.05). Vitamin A injection did not change the protein kinase B/mammalian target of rapamycin and myostatin signaling (P > 0.05). Moreover, vitamin A upregulated the expression of PAX7 (P < 0.05) and the myogenic marker genes including MYOD and MYOG (P < 0.01). The skeletal muscle-derived mononuclear cells from vitamin A-treated lambs showed higher expression of myogenic genes (P < 0.05) and formed more myotubes (P < 0.01) when myogenic differentiation was induced in vitro. In addition, in vitro analysis showed that RA promoted myogenic differentiation of the skeletal muscle-derived mononuclear cells in the first 3 d (P < 0.05) but not at the later stage (P > 0.05) as evidenced by myogenic gene expression and fusion index. Taken together, neonatal intramuscular vitamin A injection promotes lamb muscle growth by promoting the myogenic potential of satellite cells.