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Medium-chain monoglycerides (MG) have been reported to affect the productive performance, gut microbiota and health of broiler chickens reared in ideal experimental conditions at home and abroad. However, the effects of MG on performance, intestinal development and gut microbiota of chickens in large-scale farms during different feed stages remain unknown. The present study was conducted on a modern farm with a total of 12,000 yellow feathered broiler chicks that were randomly allotted to 2 groups (1000 chicks/replicate, 6 replicates/group) for a 70-day trial. The control group (CON group) received a basal diet, and the treated group (MG group) was fed a basal diet containing 300 mg/kg mixed MG. The results revealed that dietary MG significantly (P < 0.05) increased the body weight and average feed intake, but notably reduced the feed conversion and mortality of chickens in large-scale production during the starter phase. The villus height of the duodenum in the MG group at 1, 2 and 7 wk of age increased notably, and the villus height to crypt depth ratio at 1, 2, 5 and 10 wk of age was improved. Dietary MG decreased the serum insulin content of chickens at 5, 7 and 10 wk of age, and decreased the serum lipopolysaccharide at 3 and 7 wk of age. The triglyceride level of chickens at 3, 5 and 10 wk of age and the low-density lipoprotein cholesterol level of chickens at 7 and 10 wk of age in the MG group decreased notably, while the high-density lipoprotein cholesterol increased significantly. Moreover, MG supplementation selectively increased the relative abundance of genus Bacteroides(family Bacteroidaceae) and Lachnospiraceae_NK4A136_group, but decreased the content of genus Rikenellaceae_RC9_gut_group, Collinsellaand family Barnesiellaceae in the cecum of chickens at 3, 7 and 10 wk of age. Conclusively, these findings showed that dietary MG notably enhanced chicken performance, health and feed nutrient utilization at early ages by regulating gut microbiota, intestinal development and serum biochemical indices.

As the population ages, the pursuit of extended lifespan and improved healthspan has gained significance. This study assessed the anti-aging properties of sea cucumber peptides derived from Acaudina leucoprocta(SCP) through microbiome and metabolome analyses in Drosophila melanogaster. Supplementation with 0.2%, 0.4%, 0.6%, and 0.8% of SCP significantly boosted the survival rate and extended the mean and median lifespans in both male and female flies. Among them, 0.6% SCP treatment extended the mean and median lifespans of male flies by 21.01% and 25.48 %, which showed the most prominent lifespan-prolonging effect. Moreover, SCP enhanced the healthspan of male flies by improving their climbing ability, intestinal barrier function, and memory performance. Additionally, SCP boosted the survival rate of flies subjected to oxidative, starvation, and heat stress, with no significant effect observed under cold stress conditions. The lifespan-extension effect of SCP remained unaffected by dietary restriction, but was partially inhibited when provided with antibiotic-supplemented food. SCP modulated microbiota with increased the abundance of Lactobacillus, Rhodococcus, and Propioniciclavaand decreased Acetobacter, Achromobacter, Pedobacter, and norank_f_Saprospiraceae, which closely related with lifespan-extension effect. Furthermore, SCP induced a metabolic profile in 30-day-old flies more akin to that of 3-day-old flies, impacted 13 significantly regulated metabolites associated with amino acid, nucleotide, and carbohydrate metabolisms. These findings highlight SCP intervention as a promising anti-aging strategy.

Given the growing worldwide focus on bone health, exploring safe and effective interventions to maintain skeletal integrity is imperative. This study involved intragastrically administering hydrolyzed egg yolk powder (HEYP) to 3-week-old Sprague‒Dawley (SD) rats over a period of six weeks. The effects of HEYP on bone growth and the underlying mechanisms were comprehensively investigated by evaluating known bone growth parameters, bone microstructural features, serum biochemical parameters, the metabolome, and the gut microbiome. HEYP administration significantly increased longitudinal bone growth while concurrently increasing the serum levels of osteogenic growth factors. Subsequent metabolomic analyses revealed significant alterations in the serum levels of metabolites associated with bone metabolism, highlighting the profound impact of HEYP on amino acid metabolism. Additionally, gut microbiome analyses revealed HEYP-induced changes in the gut microbiota (GM), identifying key bacterial genera, including Blautia, Parasutterella, Marvinbryantiaand Prevotella, that were potentially beneficial for bone formation and linked to bone growth. Correlation analyses further highlighted associations between significantly altered serum metabolites, distinct bacterial genera, and bone growth-related parameters. These findings emphasized that HEYP might promote bone growth through various pathways, offering insights into the potential development and application of functional foods that promote bone health and potentially aid in osteoporosis prevention.

Hepatoprotective peptides are potential therapeutic substances used for curing and protecting the liver from diseases. In addition to the nutrition value, evidence indicates that these peptides have a potential role in the body's physiological process. Despite wider practicability of these peptides, their molecular structures still need to be further elucidated, to analyze and better understand its mechanism of action. Their structural variations are mainly attributed to their ultimate processing of different hydrolysates. This study summarizes potential sources, production, structure, and sequence of hepatoprotective peptides, specifically emphasizing on their protective mechanism(s) against alcoholic- and non-alcoholic liver disease. This study also describes an interaction between intestinal microbiota and hepatoprotective peptides. Preliminary research indicated that treatment with bioactive peptide promotes the balance of gut microbiota (GM), mitigates liver inflammation in mice, reverses neutrophil phagocytosis in individuals with alcoholic cirrhosis, and improves liver enzyme levels in excessive drinkers. Besides, it describes hepatoprotective peptides mechanism of action on intestinal microbiota. This work intends to add knowledge in the field of peptides and their impact on hepatoprotection.

Glycerol monolaurate (GML), a member of medium-chain α-monoglycerides (MG), is proved to be beneficial for productive performance, feed efficiency, and health of broilers based on recent research. The present study aims to evaluate the effect of MG mixture rich in GML and glycerol monodecanoate on performance, intestinal development, serum parameters, carcass yield, and muscle composition in broilers. A total of 528 chicks were weighed and randomly assigned into 4 groups (22 chicks/replicate, 6 replicates/group) for a 56-d experiment. The control group received a basal diet containing 0 mg/kg MG (CON), and the treated groups fed basal diets containing 300 (MG300), 450 and 600 mg/kg MG. The results revealed that the BW (P < 0.05), ADG, and ADFI were notably increased in MG-containing groups during the finisher phase compared with the CON group. Remarkable intestinal improvements were observed in the duodenum and jejunum of MG-treated groups, but no statistical differences were obtained. Dietary MG significantly (P < 0.05) increased the serum high-density lipoprotein cholesterol, total protein, and superoxide dismutase content in broilers. Inclusion of 300 mg/kg MG in diet increased the eviscerated yield (P = 0.066), leg muscle (P < 0.01) and breast muscle yield (P = 0.083), and improved the fresh meat quality with reduced drip loss (P < 0.01) and pH decline (P < 0.01) compared with the CON group. Moreover, the saturated fatty acid (P = 0.073), flavor amino acid (P < 0.05), and total amino acid (P < 0.05) content was notably higher in the muscle of the MG300 group than that in the CON group. In summary, these findings revealed that mixed MG can be used as an effective and novel feed supplement to improve productive performance and quality of broilers.

The present study aimed to evaluate the effects of medium-chain α-monoglycerides (MG) on productive performance, egg quality, serum biochemical indices, and gut microbiota in laying hens. A total of 252 40-wk-old Hy-Line Brown laying hens were randomly allotted into two groups (21 hens per replicate, 6 replicates per group) and fed with a basal diet (CON group) or a basal diet containing 300 mg/kg of MG (MG300 group). The eggs laid were recorded daily on a replicate basis, and egg quality was measured at 48, 56, and 64 wk of age. At the end of this trial, three randomly selected hens from each replicate were slaughtered, and the serum and cecal digesta were collected for analysis of serum biochemical indices and sex hormones and gut microbiota composition determination. The results revealed that the laying rate was significantly (P< 0.05) increased in the MG300 group, and the feed conversion ratio was decreased (P < 0.01) during 40–64 wk of age. The eggshell strength at 56 wk of age and eggshell thickness at 56 and 64 wk of age were significantly (P < 0.05) increased in the MG300 group. In addition, dietary MG significantly (P < 0.05) increased levels of serum follicle-stimulating hormone, luteinizing hormone, estradiol, glucose, Ca, serum total cholesterol, triglycerides, and high-density lipoprotein cholesterol, but decreased the lipopolysaccharide level. Notably, MG supplementation increased (P< 0.05) the relative abundance of genera Lachnospiraceae_NK4A136_group, Romboutsia, Syntrophomonas, Victivallis, Ruminiclostridium_6, and Family_XIII_UCG_001(P < 0.01) and simultaneously decreased the abundances of Proteobacteria, Faecalibacterium, Alistipes, Cerasicoccus, Schlegelella, and Treponema_2.Spearman's correlation analysis indicated that the differentiated genera were significantly associated with the serum biochemical indices and sex hormone. In summary, the present study revealed that dietary supplementation with MG can improve productive performance and egg quality by modulating gut microbiota, suggested that MG may act as an efficient feed supplement in aged hens.

The main purpose of the present study was to investigate the browning effect of 6-gingerol (6G), one of the main functional compounds in the ethyl acetate extract of ginger (ginger ethyl acetate fraction, GEF), and its underlying mechanisms. In this study, we first discovered that GEF stimulated brown adipocyte differentiation by upregulating the expression levels of browning-specific transcription makers (UCP1, PRDM16, and PGC-1α), thereby reducing lipogenesis transcriptional regulator (C/EBPα) expression in 3T3-L1-differentiated adipocytes. Then, 6G (47.81 ± 0.62 mg/g) was identified as one of the main functional compounds in GEF using high-performance liquid chromatography. 6G promoted adipocyte browning, as evidenced by an increase in some brown/beige fat-specific genes (PGC-1α, Cidea, Prdm16, Cited1, SIRT1, Tmem26, and Ucp1) and proteins (UCP1, CEBP/β, PGC-1α, and PRDM16) expression levels. Moreover, 6G greatly improved mitochondrial respiration and energy metabolism by upregulating the expression levels of some mitochondrial biogenesis markers (Tfam, Nrf1, SIRT1, and p-AMPK/AMPK) and increasing the uncoupled oxygen consumption rate of protons leaked in 3T3-L1 cells. Comparison of the experimental results obtained with an inhibitor (dorsomorphin) and an activator (5-aminoimidazole-4-carboxamide ribonucleotide) suggested that the 6G-associated regulation of the energy metabolism effect was mediated partly through the AMPK signaling pathway. This study provides new insight into the promotion of fat browning and regulation of lipid metabolism by 6G and suggests that 6G likely has potential therapeutic effects on obesity.

Butyric acid and lauric acid are short chain and medium chain fatty acids, respectively. Their anti-inflammatory and lipid metabolism regulating functions are reported, while their structural lipid functional activity still needs to be verified. In this study, obese mice induced by high-fat diet (HFD) were used to investigate the effects of butyric-lauric acid structural lipid (SBL) on inflammation and small intestinal microbes. In vivo, although SBLdid not significantly reduce body weight (P > 0.05), it might have a positive effect on mouse liver health. Compared to the corresponding physical mixture (MBL), SBLtreatment could better downregulate hepatic TNF-α, IL-1βand IL-6expression and stimulate IL-10expression. Similarly, compared with the HF group, the liver IL-1β and TNF-α levels in the LSBLtreatment group decreased by 15.2% and 22.9%, respectively (P < 0.05). In addition, HSBLtreatment increased the content of IL-10 in the liver of obese mice (increased by 26.8%, P < 0.05). Moreover, SBLmore significantly upregulated Bifidobacteriumabundance and inhibited the high-fat diet-induced increase in Faecalibaculumabundance. In all, SBLameliorated HFD-induced liver inflammation and these improvements are closely associated with its positive effects on small intestinal microbiota. This study aimed to provide support and new perspectives for the development and functional exploration of short-and medium-chain fatty acid structural lipids.

Constipation is a widespread gastrointestinal issue. This work aimed to investigate anti-constipation capacities of 9 novel strains of Lactobacillusand Bifidobacteriumusing in vitroexperiments for initial exploration and then further confirmed through in vivomodels including both zebrafish and mice. The in vitroexperimental results revealed that L. plantarumT34 and L. paracaseiYJ10 could survive under the simulated gastrointestinal tract conditions as standard strain L. rhamnosus(LGG) did. L. paracaseiYJ10 and L. caseiLc showed the highest 22 h's aggregation rate. L. rhamnosusW1 showed the highest coaggregation rate with E. coliand with S. aureus. T34 had the largest inhibition zones against S. typhimuriumand L. Monocytogenes.The highest adhesion rates with HT-29 cells were L. caseiLc, LGG and W1. Principal component analysis (PCA) and Fuzzy synthetic evaluation (FSE) were performed to evaluate the in vitroprobiotic potential of the 9 strains comprehensively. The order of scores on in vitroprobiotic property from high to low was as follows: T34, W1, YJ10, Lc, LGG, SLT, SS, Lb, YN4 and LQSQ. The subsequent in vivoexperiment of zebrafish further confirmed T34, W1 and YJ10 have better anti-constipation ability compared to other strains. Furthermore, T34 relieve loperamide-induced constipation by promoting gut transit in the mice model. In conclusion, T34 was demonstrated to prevent constipation and may be suggested for use in food and pharmaceutical goods.

The ethylcellulose/gelatin solutions containing various concentrations of zinc oxide (ZnO) nanoparticles were electrospun, and the resultant nanofibers were characterized by scanning electron microscopy, energy dispersive X-ray, X-ray photoelectron spectrometer, X-ray diffraction, Fourier transform infrared spectroscopy, mechanical testing, water contact angle, and water stability. Results indicated that ZnO nanoparticles acting as fillers interacted with polymers, resulting in the enhanced surface hydrophobicity and water stability of nanofibers. The antibacterial assay showed a concentration-dependent effect of ZnO on the viabilities of Escherichia coliand Staphylococcus aureus. Notably, the antimicrobial efficiency of the 1.5 wt % ZnO-containing fibers against Staphylococcus aureuswas 43.7% but increased to 62.5% after UV irradiation at 364 nm, possibly due to the significantly increased amounts of intracellular reactive oxygen species. These results suggested that the ZnO-containing nanofibers with excellent surface hydrophobicity, water stability, and antimicrobial activity exhibited potential uses in food packaging.

In this study, chitosan/poly(ethylene oxide) (PEO)/lauric arginate (LAE) composite nanofibrous films were fabricated via electrospinning. The addition of LAE did not change the physical properties of chitosan/PEO in acetic aqueous solutions, but increased the fluorescent intensity of chitosan by electrostatic interactions, resulting in uniform and bead-free nanofibers with an average diameter of 150 nm. The Fourier transform infrared spectra and thermal analysis indicated that the LAE molecules were homogeneously dispersed within the chitosan/PEO nanofibers. The formation of electrostatic and hydrogen bonding interactions induced by the LAE addition changed the inter- and intramolecular interactions between PEO and chitosan and further affected the mobility of the polymer molecules, leading to the increased crystallinity and decreased melting point. The hydrophilicity of the nanofibrous films was significantly increased by the incorporation of LAE, as indicated by the decreasing water contact angle from 39° to 10°. Meanwhile, the chitosan/PEO/LAE nanofibrous films showed LAE concentration dependent antimicrobial activity against Escherichia coliand Staphylococcus aureus, suggesting enhanced antimicrobial activity. The fluorescent staining experiments demonstrated that the antimicrobial mechanism of the nanofibrous films was cell membrane damage.

Constipation has become more common in recent decades, affecting people’s quality of life. Natural bioactive substances could be effective in alleviating constipation. The goal of this study was to assess and compare the efficacy of low and high hydrolysis degree wheat peptides (WP-L and WP-H) at a dose of 1 mg/g.bw on relieving loperamide-induced constipation in mice and the underlying mechanisms. Here, we proved that both WP-L and WP-H treatment showed constipation relieving effect by improving defecation function and small intestinal propulsion rate while the different hydrolysis degree did not affect the efficacy of WP. Moreover, WP-L and WP-H could balance the secretion of excitatory and inhibitory factors, and enhance the activity of antioxidant enzyme (SOD and GSH-Px). Importantly, both of them could accelerate intestinal motility, improve water-salt metabolism and intestinal barrier, and ameliorate oxidative stress. In summary, our findings indicated that 1 mg/g.bw WP-L and WP-H effectively relieved loperamide-induced constipation by affecting multiple targets in mice.

Medium- and long-chain triglycerides (MLCT) have been widely concerned. In this study, lauric acid (La) was incorporated into flaxseed (Linum usitatissimumL.) oil (FO, aLna-riched) by lipase-catalyzed acidolysis. The synthesis efficiency of four immobilized lipases (Lipozyme RM IM, TL IM, AOAB8 and Novozym 435) was compared, while Lipozyme AOAB8 reaction for 2 h was the best scheme. Further, the optimal conditions for synthesizing structural lipids (aLSLs, rich in aLna and La) were obtained by response surface methodology: the reaction temperature was 44.6 °C, substrate molar ratio was 12 (La/FO), and enzyme loading was 9.11%. After purification by molecular distillation, the triglycerides (TAGs) of aLSLs contained 75.35% of TAGs with α-linolenic acid (aLna), 40.82% of MLCTs with La, and 26.75% (65.53% of the total MLCTs) of TAGs with both aLna and La (ɑ-MLCT). Meanwhile, the content of La and aLna in aLSLs were 22.38% and 39.78%, respectively. The chemical structure and microstructure of aLSLs were similar to FO. In addition, the thermo-oxidative decomposition behavior of aLSLs was significantly improved compared with physical mixture oil. This improvement might be related to the insertion of La and the structure of TAG. In all, this study firstly developed a novel thermo-oxidatively stable MLCT with high levels of aLna and La.

Background: Propeptides of lipases have been demonstrated to influence many properties of their mature proteins as intramolecular chaperones. However, the working mechanism of propeptides may be different. Objectives: The main objective of this study was to determine the role of pentapeptide prosequence of Thermomyces lanuginosus lipase (TLL) through its effect on the recombinant expression of TLL in P. pastoris, and explore the possible function mechanism with its hydrophobicity. Methods: We have synthesized a codon-optimized TLL gene coTLL with a Kex2 cleavage site “-KREAEA-” directly after the propeptide “SPIRR-”, and obtained expression vector pP-kTL through cloning it into pPIC9K. TL gene without the propeptide and pTL gene with a propeptide directed linked to mature TLL lipase, were also amplified and cloned into pPIC9K to obtain the expression vector pP-TL and pP-pTL. pTL-P and pTL-VP gene variants with mutation in pentapeptide prosequence of TLL were obtained used the site-directed mutation with the pP-pTL plasmid as the template. The recombinant proteins were expressed in P. pastoris GS115. Lipase activity was determined by a spectrophotometric method previously reported using para-nitrophenyl palmitate (pNPP) as the substrate and the thermostability of lipases of TLL was analyzed by incubating at different temperatures (70°C and 80°C) for 5h and molecular dynamics simulation. Results: The average lipase activity of recombinant strains GS-pTL reached 434.32 U/mL, higher than that of GS-TL 377.71 U/mL. The fermentation result of the recombinant strains with modified propeptide showed that the extracellular lipase activity of GS-pTL-VP variant reached 483.29 U/mL, increasing by 11.27% compared with that of GS-pTL (434.32 U/mL). Further analysis performed on the lipase stabilities with propeptide variants by molecular dynamics simulation showed that the RMSD of variant pTL-VP was similar to that of pTL. Conclusion: This study revealed that the propeptide “SPIRR-” sequence is beneficial for enhancing TLL expression. In addition, the function of TLL propeptide was identified to be related to its hydrophobicity, implying that propeptide might play a role in assisting the formation of the hydrophobic protein core and accelerate the protein folding process. This work inspired us to attach more emphasis on the propeptide of other lipases for improving their recombinant expression, structure folding and enzymatic properties.

Constipation is a common gastrointestinal problem. This study aimed to identify a combination of prebiotics and wheat peptides (WP) to promote intestinal peristalsis and explore the underlying mechanism. First, we screened four prebiotics (galacto-oligosaccharides (GOS), fructose oligosaccharides (FOS), inulin (IN), and resistant dextrin (RD)) using a zebrafish model and found that GOS was the most synergistic with WP. We then tested the GOS-WP combination in a mouse model of loperamide (Lop)-induced constipation and found that it significantly improved constipation symptoms. Specifically, GOS-WP shortened the first black stool time, increased the number of defecation particles, accelerated small intestinal propulsion rate, and increased fecal water content. GOS-WP also normalized the colonic water transport system by inhibiting colonic AQP4/8expression. Additionally, GOS-WP reversed Lop-induced morphological damage in the ileum and colon, increased colonic barrier, and reduced inflammation in mice. Furthermore, 16S rRNA gene sequence analysis showed that GOS-WP reversed the gut microbiota structure changes induced by Lop. GOS-WP significantly increased butyric acid and decreased BCFA levels, indicating improved gut health. Our findings suggest that the GOS-WP combination has potential as a novel intervention for constipation relief. This study highlights the potential of combining prebiotics and WP as a therapeutic strategy for constipation.

Glycerol monolaurate (GML) has the potential of antibacterial and growth-promoting in food. Recent research reported that GML could significantly increase the body index of fish, but whether GML can exert growth-promoting ability via regulating bone metabolism remains unknown. This study explored the effects of GML on the bone growth of fish models and its mechanisms by analyzing the expression of insulin-like growth factor 1(IGF-1). The micro-CT scanning results showed that the dietary supplementation of 750 mg/kg GML significantly increased the body length and bone mineral density, and improved the bone microarchitecture parameters of the caudal vertebrae of zebrafish (Danio rerio). The centrum part of the vertebra was easier to respond to the regulatory effect of GML than the neural arch and the haemal arch. Mechanistically, the content of serum IGF-1, the expression of osteogenic differentiation-related genes, and genes involved in IGF-1/PI3K/Akt signaling pathway were significantly increased. The liver proteomic analysis showed the consistent results that GML supplementation could up-regulate the expression of hepatic IGF-1, and that energy metabolism and growth and development-related metabolic pathways were implicated in the GML bone growth boosting effects. These findings highlight that GML could be used as a functional food additive to improve bone growth and health.

•This study evaluated the nutritional effects of dietary GML using zebrafish as a model animal.•Dietary supplementation of GML significantly improved the weight gain and feed utilization in zebrafish.•GML showed promising potentials by increasing the intestinal enzymes and anti-inflammatory activities.•GML supplementation positively altered and reconstructed the gut microbiota of zebrafish.

•A first nutritional study on the effect of dietary GML in the field of aquaculture.•Treatment with 700 and 1050 mg/kg GML significantly improved the growth, lipase and protease activities in intestine.•The addition of 350 and 700 mg/kg GML significantly enhanced the antioxidant and antibacterial capacity of shrimp.•The optimal dietary supplementation dosage is 700 mg/kg in diet.

Obesity and associated metabolic disorders are worldwide public health issues. The gut microbiota plays a key role in the pathophysiology of diet-induced obesity. Glycerol monolaurate (GML) is a widely consumed food emulsifier with antibacterial properties. Here, we explore the anti-obesity effect of GML (1,600 mg/kg of body weight) in high-fat diet (HFD)-fed mice. HFD-fed mice were treated with 1,600 mg/kg GML. Integrated microbiome, metabolome, and transcriptome analyses were used to systematically investigate the metabolic effects of GML, and antibiotic treatment was used to assess the effects of GML on the gut microbiota.