The 2023 iteration of the Global Burden of Diseases, Injuries, and Risk Factors Study (GBD) estimated prevalence, incidence, and health burden for 375 diseases and injuries, including 12 mental disorders. We assess past, current, and emerging trends in the prevalence and burden of mental disorders across sexes and age groups, for 21 regions, 204 countries and territories, and by Socio-demographic Index (SDI) quintile, from 1990 to 2023. Mental disorders included in GBD 2023 were anxiety disorders, major depressive disorder, dysthymia, bipolar disorder, schizophrenia, autism spectrum disorders, conduct disorder, attention-deficit hyperactivity disorder, anorexia nervosa, bulimia nervosa, idiopathic developmental intellectual disability, and a residual category of other mental disorders. A literature review identified epidemiological data for each disorder. These were analysed via a Bayesian meta-regression to estimate prevalence by disorder, sex, age, location, and year. Disorder-specific prevalence was multiplied by disability weights representing the severity of health loss associated with each disorder to estimate years lived with disability (YLDs). Deaths due to anorexia nervosa were assessed with a Cause of Death Ensemble modelling strategy to estimate deaths by sex, age, location, and year, and then multiplied by the standard life expectancy at age of death to estimate years of life lost (YLLs). YLDs equalled disability-adjusted life-years (DALYs) for all mental disorders except anorexia nervosa (the only mental disorder considered as an underlying cause of death in GBD), for which DALYs represented the sum of YLDs and YLLs. We presented prevalence, deaths, YLDs, YLLs, and DALYs as counts, age-specific rates per 100 000 population, and age-standardised rates per 100 000 population. We estimated 1·17 billion (95% uncertainty interval 1·06-1·31) prevalent cases of mental disorders globally in 2023, equivalent to an age-standardised prevalence rate of 14 210·7 cases (12 849·5-15 940·1) per 100 000 population. These estimates represented a 95·5% (75·0-121·2) increase in prevalent cases and 24·2% (11·4-41·4) increase in age-standardised prevalence rate between 1990 and 2023. All mental disorders showed increases in prevalent cases between 1990 and 2023, while notable increases were seen in age-standardised prevalence rates for anxiety disorders, major depressive disorder, dysthymia, anorexia nervosa, bulimia nervosa, schizophrenia, and conduct disorder. There were an estimated 171 million (127-228) DALYs due to mental disorders globally across sex and age in 2023, equivalent to an age-standardised DALY rate of 2070·5 DALYs (1519·1-2750·5) per 100 000 population. Mental disorders contributed to 6·1% (4·8-7·6) of all-cause DALYs in 2023, making them the fifth leading cause of global DALYs (up from 12th in 1990). DALYs were almost entirely composed of YLDs. Mental disorders were the leading cause of YLDs in 2023 (up from second in 1990), explaining 17·3% (14·8-20·6) of all-cause global YLDs. Leading causes of mental disorder DALYs were anxiety disorders (ranked 11th among the 304 diseases and injuries at Level 4 of the GBD cause hierarchy), major depressive disorder (15th), and schizophrenia (41st). Globally in 2023, mental disorder age-standardised DALY rates were higher among females (2239·6 [1643·7-3014·1] per 100 000) than among males (1900·2 [1399·8-2510·8] per 100 000), and peaked in the 15-19 years age group (2617·3 [1850·6-3696·8] per 100 000). All locations showed increased mental disorder DALY rates in 2023 compared with 1990, ranging across countries and territories from 1302·4 (952·7-1683·7) per 100 000 in Viet Nam to 3555·8 (2661·9-4715·0) per 100 000 in the Netherlands. Across SDI quintiles, DALY rates ranged from 1853·0 (1352·1-2469·3) per 100 000 for middle SDI to 2184·1 (1606·1-2890·3) per 100 000 for high SDI. A significant health burden was imposed by mental disorders in all countries and territories in 2023, irrespective of the health resources available. In some instances, this burden has increased over time and is unevenly distributed across populations. Stronger surveillance systems, particularly in low-income and middle-income countries, are required. Additionally, we need more coordinated and inclusive policies to reduce the burden through early treatment and prevention, tailored to sex and age differences across locations. Responding to the mental health needs of our global population, especially those most vulnerable, is an obligation, not a choice. Gates Foundation, Queensland Health, and University of Queensland.
Improving milk yield and feed efficiency is pivotal for climate-smart dairy systems, as rumen mediated fermentation governs energy and nitrogen utilization and thereby greenhouse-gas emission intensity. Soybean isoflavones (SIF) may modulate rumen fermentation, yet their effects on rumen function, microbiome features, host endocrine/metabolic responses, and lactation performance-particularly across cows with divergent milk-yield phenotypes-remain unclear. Fifty‑six lactating Holstein cows (28 high‑yield cows, HY; 28 low‑yield cows, LY) were divided into two categories by milk yield. Within each yield category, cows were randomly assigned to one of two dietary treatments: a basal diet (Control) or the basal diet supplemented with SIF at 0.01% of dry matter. This yielded a 2 × 2 factorial design with four experimental groups (n = 14 per group): high‑yield control (HCON), high‑yield SIF (HSIF), low‑yield control (LCON), and low‑yield SIF (LSIF). SIF increased milk yield by 8.75% and improved fat-corrected milk (+ 7.20%), dry matter intake (+ 3.20%), and feed efficiency (+ 3.26%), with larger gains in HY cows (milk yield + 8.89%; feed efficiency + 4.55%). Rumen fermentation shifted toward a more energetically favorable profile, with lower acetate (- 2.70%), higher propionate (+ 4.55%), and a reduced acetate-to-propionate ratio (- 7.02%), accompanied by increased microbial crude protein (+ 21.53%) without changes in pH or NH3-N. SIF altered endocrine status irrespective of phenotype, increasing estradiol and progesterone while decreasing prolactin and growth hormone, and reduced blood ALP, lactate, and triglycerides. Metagenomics indicated phenotype-dependent microbial and functional responses to SIF: HY cows showed enrichment of taxa (e.g., Caudoviricetes sp., Eubacterium sp., and Butyrivibrio sp.) associated with amino-acid, cofactor metabolism and propionate pathways, whereas LY cows exhibited enrichment of Prevotella sp. and Bacteroides sp. with functions favoring carbohydrate degradation. The HCON group exhibited greater abundances of Prevotella sp. and Hallella spp. with enhanced carbohydrate degradation functions, whereas the LCON group was enriched in Ruminococcus sp. and Methanobrevibacter sp., associated with methane metabolism. In conclusion, this study highlights the potential of SIF supplementation to improve lactation efficiency, modulate rumen microecology and endocrine function in dairy cows. These findings establish a theoretical framework for achieving efficient and precise feeding management on large-scale dairy farms.
Citrulline (Cit), an effective precursor of arginine (Arg), escapes hepatic catabolism to be almost completely absorbed into the systemic circulation, thereby being efficiently converted to Arg in the kidneys to enhance its systemic bioavailability. This study investigated the effects of dietary Cit supplementation on lactation performance in sows, as well as the underlying mechanisms related to intestinal health in their suckling piglets, using multi-omics analyses. Dietary Arg and Cit supplementation significantly increased average daily feed intake of lactating sows. Milk fat content and plasma nitric oxide (NO) concentration increased significantly in the Arg group and the 40%Cit group (P < 0.05), while milk threonine content increased slightly (P = 0.084). Consequently, the average daily gain of suckling piglets over the 21-day lactation period was also significantly improved. Furthermore, maternal 40%Cit supplementation improved the intestinal health of offspring by enhancing jejunal morphology and upregulating the expression of the tight junction protein occludin (P < 0.05), indicating a strengthened intestinal barrier. Mechanistically, this was achieved by activating the mTOR/S6 pathway in the piglets' jejunum. Maternal 40%Cit supplementation upregulated the expression of proteins related to mitochondrial fusion and fission (MFN2 and MFF, P < 0.05), and the protein expression of OPA1 showed an increasing trend (P = 0.097), indicating the structural and functional status of mitochondria was improved. Maternal 40%Cit supplementation also modulated the gut microbiota of piglets, increasing the abundance of beneficial bacteria (Lachnoclostridium). Metabolomic analysis of sow milk identified 58 differential metabolites. Among these metabolites, palmitic acid levels were significantly increased and positively correlated with the abundance of Lachnoclostridium in the intestine (P < 0.05). Dietary Cit supplementation enhanced sow lactation performance and improved intestinal barrier function in their offspring via activation of the jejunal mTOR/S6 pathway and improved mitochondrial structure and function in the piglet jejunum. These benefits were further supported by modulation of the gut microbiota and alterations in the milk fat and metabolome, ultimately promoting piglet growth.
Breast muscle yield is a key economic trait in broilers, directly affecting carcass value and profitability, and has been significantly improved by intensive selection and exploiting heterosis through crossbreeding. Our previous work showed that synergy between the gut microbiota and host genome underlies breast muscle heterosis in crossbred progeny (CR) derived from Cornish (CC) and White Plymouth Rock (RR) lines. However, the molecular mechanisms by which the gut microbiota contributes to heterosis in breast muscle yield remain poorly understood. Here, we integrated cecal microbiome, metabolome, and transcriptome data from 266 birds at 42 days of age to elucidate the potential gut microbiota-mediated molecular mechanisms underlying breast muscle yield heterosis. To assess whether heterosis extends beyond productive traits to the gut microbiota and their metabolites in broilers, we compared the cecal microbial and metabolic profiles of CR with those of their parental lines. The gut microbiota of CR were clearly distinct from those of both parental lines and exhibited heterosis characteristics, with 88 genera displaying heterotic patterns that collectively accounted for approximately 85% of the total microbial abundance. Heterosis was also evident in the cecal metabolites of CR birds. Differential abundance analysis across groups identified 868 cecal metabolites, and abundance-pattern classification showed that approximately 75% exhibited nonadditive patterns in the crossbred progeny. These nonadditive metabolites were predominantly host-microbiota co-metabolites and were mainly enriched in amino acid and lipid metabolic pathways. Importantly, seven of the nine genera previously identified in association with breast muscle yield exhibited heterosis in the crossbred progeny. At the metabolomic level, yield-associated genera were linked to a distinct set of 35 cecal metabolites, dominated by sphingolipids, ether-linked phospholipids, and acyl-homoserine lactones. These metabolites formed coordinated associations with the expression of 269 host genes, which were functionally enriched in MAPK signaling and focal adhesion pathways. These findings suggest that heterosis exists not only in productive traits but also in gut microbiota and their metabolites, the latter in turn contributed to breast muscle yield, which offers valuable guidance for elucidating the molecular basis of heterosis in animals.
High levels of zinc oxide (ZnO) and copper sulfate are widely used as alternative growth promoters in postweaning pig diet. However, excessive exposure to these metals may drive co-selection for heavy metal (HMR) and antibiotic resistance (AMR). Nursery diets also contain abundant iron to offset the low bioavailability of plant-derived iron, yet how dietary iron influence gut dysbiosis and microbial resistance in postweaning pigs remains unclear. This exploratory study examined the effects of dietary iron and metal-based growth promoters on the fecal resistome of postweaning pigs using shotgun metagenomics and whole-genome sequencing (WGS). Fifty weanling pigs were stratified and randomly assigned to five dietary treatments for 24 d. Experimental diets included a control diet (Con) containing 25, 139, and 141 mg/kg of Cu, Fe, and Zn, respectively, a low-iron diet (LFe, 19 mg Fe/kg), a high-iron diet (HFe, 1,219 mg Fe/kg), a high-copper diet (HCu, 257 mg Cu/kg), and a high-zinc diet (HZn, 2,631 mg Zn/kg, including 2,490 mg Zn/kg from ZnO). All pigs were orally administered with F18 enterotoxigenic Escherichia coli (ETEC) on d 13-16. Metagenome sequencing were performed on d 24 fecal DNA (n = 24) to identify HMR genes (BacMet Predicted database) and AMR genes (CARD database). Functional annotation was performed using HUMAnN3. Whole genome sequencing (WGS) was conducted on 120 E. coli isolates from fecal cultures on d 1, 12, and 24, and AMR and virulence genes were identified from contig assemblies using ABRicate. Dietary metal treatments significantly altered β-diversity of HMR genes compared with Con, with HZn differing from both HCu and LFe (P < 0.05). Fecal iron levels correlated with sodB (ρ = 0.64, P = 0.075), an iron-containing superoxide dismutase, while fecal copper levels correlated with pcoC (ρ = 0.66, P = 0.075), a plasmid-mediated copper resistance gene. Across metagenomes, 172 AMR genes were identified, dominated by glycopeptide and tetracycline resistance. While dietary iron had minimal effects on fecal AMR profile, HZn induced the largest shifts in resistome, including increases of ant(9)-la, conferring aminoglycoside resistance on mobile genetic elements, and adeF, encoding a multidrug efflux pump (P < 0.05). Functional profiling revealed enrichment of carbohydrate metabolism pathways in HZn group (P < 0.05). WGS of E. coli isolates showed distinct AMR profiles under HZn on d 24 and distinct virulence profile under LFe on d 12, exhibiting increased prevalence of exotoxin and T3SS genes (P < 0.05). Dietary iron restriction enhanced E. coli virulence genes, whereas excessive ZnO induced the most pronounced changes in the gut resistome and microbial metabolism, highlighting a risk for AMR co-selection and marked influence on gut microbiota.
Gamma-aminobutyric acid (GABA) influences metabolic homeostasis, immune function, and lactation performance. Typically, GABA is administered exogenously, but this approach is limited by intake variability and cost. Alternatively, silage inoculated with high GABA-producing Lentilactobacillus buchneri YM9 enriches the silage with GABA and ensures more uniform dietary delivery. However, the effects of silage-based GABA enrichment and delivery on ruminant performance, immunity, and health remain unclear. Hence, we conducted a feeding trial with 36 dairy goats assigned to three TMR treatments containing alfalfa silage: 1) CK (uninoculated control), 2) AH35 (inoculated with non-GABA-producing Lent. buchneri AH35), and 3) YM9 (inoculated with GABA-producing Lent. buchneri YM9). Feed intake and lactation performance, rumen fermentation and microbiota, blood GABA and lactation hormones, blood cytokines and interleukins as well as milk GABA and mammary gland gene expression were evaluated. YM9-fed goats had lower DMI (1.66 kg/d) compared with CK (1.84 kg/d) and AH35 (1.84 kg/d) but maintained milk yield comparable to CK and higher than AH35 (P < 0.05). GABA intake, and milk yield, fat, protein, lactose, total solids, and total nitrogen per kilogram DMI were higher in YM9 (P < 0.05). Milk GABA was greater in YM9 (152 μmol/L) than CK (114 μmol/L) and AH35 (111 μmol/L) (P < 0.01). Serum GABA (1.44 μmol/L), prolactin (7.65 ng/mL), and oxytocin (4.75 pg/mL) were elevated in YM9 (P < 0.05). Immunoglobulins were higher in AH35 overall, but YM9 exceeded CK (P < 0.05), with cytokine profiles in the YM9 group reflecting moderate pro-inflammatory activation. YM9 upregulated GSR and downregulated NOX4, TNF, and IFNG (P < 0.05). Microbiota analysis showed comparable alpha diversity between YM9 and CK with Prevotella dominance in the YM9 group, and significant correlations among GABA, microbial taxa, hormones, and mammary gland genes. Feeding dairy goats with GABA-enriched silage was associated with improved feed and lactation efficiency. It also enhanced lactation hormones, immune responses, and expression of antioxidant-related gene alongside reduced inflammatory related genes. These findings provide alternative approach for dietary GABA delivery to ruminants for enhanced productivity and health, although further studies are required to verify the mechanisms.
Subclinical mastitis (SCM) is a major constraint in dairy production and is driven by complex host-pathogen interactions. Although transcriptional responses associated with SCM have been widely investigated, the epigenetic mechanisms that stably regulate these programs remain less well characterized, particularly in crossbred cattle populations. This study aimed to characterize DNA methylation-based regulatory networks by integrating whole-genome methylation and transcriptome data from milk somatic cells of Vrindavani (Bos taurus × Bos indicus) cattle. Whole-genome methylation (n = 6) and corresponding transcriptome profiling (n = 6) were performed on milk somatic cells from SCM-affected and healthy control cows. Differential methylation analysis (q-value < 0.05) identified 62,940 differentially methylated cytosines (DMCs), 7,706 differentially methylated regions (DMRs), and 6,203 differentially methylated genes (DMGs), with a predominant bias toward hypomethylation in SCM. Integrative analysis using stringent thresholds for both methylation (≥ 10%) and expression change (|log₂ fold change| ≥ 1; P of GMM < 0.001) identified 1,407 differentially methylated and expressed genes (DMEGs). Functional enrichment analysis revealed 47 KEGG pathways and 30 Gene Ontology biological process terms (FDR < 0.05), primarily associated with immune signaling and inflammatory responses. In contrast, a subset of DMEGs showed methylation-associated repression of lactation- and metabolism-related genes. Selected genes were experimentally validated by qPCR, including upregulation of the inflammatory mediator S100A8 and downregulation of CSN3 (κ-casein), a key milk protein gene. These findings provide an integrated view of the DNA methylation and transcriptional landscape of SCM in milk somatic cells and demonstrate that epigenetic remodeling is associated with coordinated activation of immune pathways alongside repression of lactation-associated functions. The results contribute to understanding the molecular basis of subclinical mastitis and may inform future efforts toward biomarker development and epigenetically informed strategies for improving disease resilience in dairy cattle.
The fact that feeding pigs with probiotic-fermented agricultural by-products improves pork quality has been repeatedly demonstrated and widely applied, but the underlying mechanisms remain unclear. This study explored the effects of fermented extruded brewers' spent grain (FEBSG) on meat quality in growing-finishing pigs, as well as its regulatory mechanisms. Sixty Duroc × Landrace × Yorkshire pigs (52.25 ± 2.10 kg) were randomly assigned to five dietary treatments, in which FEBSG replaced 0, 5%, 10%, 15%, and 20% of soybean meal (SBM). The experiment spanned 10 weeks. Compared with the control, 20% FEBSG significantly increased final body weight, average daily feed intake, and average daily gain, while decreasing feed to gain ratio (P < 0.05). Both 15% and 20% FEBSG improved carcass characteristics and meat quality, including higher carcass weight, loin eye area, and intramuscular fat content, along with lower drip loss and shear force (P < 0.05). These treatments also enhanced flavor-related amino acids and unsaturated fatty acids (P < 0.05), and improved umami and sweet taste profiles. Moreover, 20% FEBSG increased muscle fiber density and reduced fiber diameter, upregulated MyHC I, MyHC IIa, PGC-1α, AMPKα1, TFAM, and SDH activity, and downregulated MyHC IIb and LDH activity (P < 0.05). Proteomic analysis identified 69 differentially expressed proteins, with enrichment in AMPK and PPAR signaling pathways. Metagenomic analysis revealed increased abundance of short-chain fatty acid-producing bacteria, including Clostridium, Lactobacillus, Prevotella, and Bartonella. Correlation analysis demonstrated associations between gut microbiota diversity and meat quality traits, as well as between dominant microbial genera and differentially expressed proteins, volatile fatty acids, muscle fiber characteristics, and the AMPK/PGC-1α/TFAM signaling pathway. Partial replacement of SBM with FEBSG positively influenced growth performance and pork quality in pigs, with the underlying mechanisms may involve the activation of the AMPK/PGC-1α/TFAM signaling pathway via the gut-muscle axis, thereby enhancing mitochondrial biogenesis, muscle development, and metabolism.
Female reproductive aging is a fundamental biological process characterized by a progressive decline in ovarian function, oocyte quality, and endocrine homeostasis, ultimately leading to reduced fertility and increased susceptibility to age-related diseases. Accumulating evidence indicates that reproductive aging is not merely a passive consequence of time but rather a tightly regulated process governed by complex genetic, epigenetic, and metabolic mechanisms. However, mechanistic dissection and translational exploration of female reproductive aging remain constrained by the limited availability of suitable animal models that faithfully recapitulate the human reproductive trajectory. In this review, we synthesize the current advances in understanding the molecular regulatory networks underlying female reproductive aging, with particular emphasis on key signaling pathways, cellular senescence, epigenetic regulation, hormonal control, and mitochondrial dysfunction coupled with oxidative stress. We highlight how the dysregulation of these interconnected mechanisms contributes to ovarian reserve depletion, follicular atresia, and declining oocyte competence across species. We propose that laying hens are a powerful and underutilized model for studying female reproductive aging. Laying hens exhibit a well-defined and highly reproducible reproductive lifespan characterized by distinct phases of peak and declining reproductive output, closely paralleling the age-related fertility decline in women. At the molecular level, hens share conserved regulatory features with humans, including hormonal signaling via the hypothalamic-pituitary-ovarian axis, age-associated oxidative stress, mitochondrial dysfunction, and epigenetic modulation of reproductive tissues. The daily ovulation cycle, measurable reproductive output, and responsiveness to metabolic and environmental interventions in hens further facilitate high-resolution and high-throughput investigations into aging-related mechanisms. By integrating evidence from human studies, mammalian models, and avian systems, this review highlights the translational value of laying hens in elucidating conserved genetic and epigenetic drivers of female reproductive aging. We discuss the current limitations and future perspectives for cross-species validation and multi-omics integration, aiming to facilitate the identification of actionable targets for delaying reproductive aging and improving female reproductive health.
Genomic prediction is widely used in pig breeding, but phenotypic prediction of complex traits such as disease resilience remains limited because genotypes alone do not capture infection-induced regulatory responses, environmental and management effects, or their interactions. Blood molecular profiles measured in young healthy pigs reflect both genetic and non-genetic influences and may improve prediction of performance under disease challenge. We evaluated whether integrating multiple blood-based omics layers with genomic data improves prediction of production and disease resilience phenotypes in pigs exposed to a polymicrobial disease challenge. Data were from 836 healthy pigs from 15 batches with transcriptomic, proteomic, and metabolomic profiles measured in blood collected at ~27 days of age, before transfer into a natural polymicrobial disease challenge at ~40 days of age. Pigs were also genotyped using a commercial 650 K marker array. We analyzed 21 traits related to growth, health scores, antibiotic treatments, mortality, feed efficiency, and carcass traits using best linear unbiased prediction (BLUP) animal models with random animal effects based on relationship matrices constructed from genomic (G), transcriptomic (T), proteomic (P), and metabolomic (M) data. Across traits, G-BLUP explained the largest proportion of phenotypic variance for most traits. However, T-, P-, or M-BLUP explained similar or greater variance than G-BLUP for several growth and health traits recorded before challenge. Adding T and/or M to G-BLUP generally increased variance explained and improved prediction accuracy for pre-challenge growth rate and health scores, and for mortality and carcass weight after challenge. Models combining G, T, and M often yielded the highest accuracies, whereas adding P did not consistently improve accuracy. For later grow-finish traits, gains from multi-omics were smaller and less consistent. Blood multi-omics profiles from healthy young pigs can improve prediction of performance and disease resilience beyond genomic data alone. Gains were greatest for traits recorded before challenge and for some resilience traits expressed soon after pathogen exposure, suggesting that pre-challenge molecular profiles capture latent resilience potential. These findings support the use of pre-challenge blood multi-omics as biomarkers for precision management and as a basis for breeding and management strategies targeting disease resilience in pigs.
Baicalin is a bioactive flavonoid from Scutellaria baicalensis Georgi with antioxidant, anti‑inflammatory and antibacterial properties. However, its bitter taste and susceptibility to ruminal degradation limit its practical use in mammals. Enteric coating technology might overcome these limitations by enabling targeted intestinal release. This study investigated effects of dietary supplementation with baicalin and coated baicalin on rumen fermentation, gastrointestinal microbiota, immune function and growth performance in Hu sheep. Thirty-six lambs with similar body weight (33.01 ± 2.68 kg) were randomly assigned to three groups (4 replicates per group, 3 sheep per replicate). The control group was fed basal diet (CON) while treatment I (BAI) and treatment II (C-BAI) groups were fed a basal diet supplemented with 0.1% baicalin and coated baicalin, respectively. After a 60-d feeding trial, baicalin and coated baicalin supplementation improved total weight gain and average daily gain compared with CON group (P < 0.05). In addition, BAI and C-BAI groups exhibited higher total antioxidant capacity (P < 0.05) and catalase activity (P < 0.05) with associated lower malondialdehyde levels (P < 0.05). Immunoglobulin G and anti-inflammatory cytokines interleukin-4 (IL-4) were also increased (P < 0.001). Notably, IgM, IL-10 and IL-4 in the C-BAI group exceeded those of the BAI group (P < 0.001). Microbiome analysis revealed that baicalin supplementation enriched abundance of beneficial bacterial taxa including Firmicutes and Lachnoclostridium (P < 0.05) and reduced potential pathogen abundance, e.g., Treponema and Ralstonia (P < 0.05). The C-BAI group also showed increased abundance of the beneficial Bradyrhizobium compared with CON (P < 0.05). Metabolomic analysis revealed that baicalin altered propionate and tyrosine metabolic pathways (P < 0.05), while coated baicalin modulated penicillin metabolism and glyceride metabolism in jejunum (P < 0.05) increasing ATP production. Overall, these results indicated enhanced nutrient metabolism and gut health in the presence of dietary baicalin. Dietary supplementation with baicalin and coated baicalin improved growth performance, antioxidant status, immunity and beneficially modulated the microbiome-metabolome crosstalk in Hu sheep. Notably, uncoated baicalin exerted more pronounced effects on growth performance and supported a role for baicalin as a potential and functional feed additive.
Cancer is a major global issue threatening the whole world, especially developing countries. Treatment of cancer using chemotherapy and radiotherapy has several consequences that negatively affect the quality of life of cancer patients. Bee products have numerous pharmacological effects and clinical impacts due to their extraordinary chemical composition. The objective of the current work is to shed light on preclinical studies and clinical trials of bee products, particularly propolis, honey, and royal jelly, with special emphasis on their role in reducing the complications of chemotherapy and radiotherapy by employing a variety of databases. The search used specific keywords, including "bee products", "propolis", "honey", "royal jelly", "cancer", "clinical trials", "radiotherapy", and "chemotherapy". Only peer-reviewed randomized controlled trials (RCTs) and published research papers were included. According to the literature review, bee-generated propolis, honey, and royal jelly have been used in animal models to reduce the adverse effects of radiotherapy and chemotherapy. Depending on the kind of cancer, different dosages and treatment times were used for certain bee products. Bee products are used in various forms, such as crude, in capsules, mouthwashes, tablets, and oils. Propolis, royal jelly, and honey are used at dosages up to 400 mg, 1 g, and 50 g, respectively. Clinical trials have further confirmed their efficacy in cancer treatment, either as standalone therapies or as supplements to conventional treatments. It is crucial to investigate the active mechanisms of these products further and to include them in additional clinical trials as potential cancer treatments.
As global demand for milk and meat increases, young ruminants need rapid gut development and sufficient mucosal immunity immediately after birth. However, their growing intestines face oxidative and microbial challenges that can harm long-term performance. This review systematically summarizes recent advances in how host genetics, early nutrition, and emerging microbiota interact to influence intestinal development and mucosal immunity under standard rearing conditions. We explore how the host regulates the gut microbiota, the microbiota's role in maintaining gut integrity, barrier function, development, intestinal cell health, mucosal immunity, and how diet modulates gut microbiota composition in young ruminants. Microbial signals promote increased villus length and better epithelial functions. Conversely, dysbiosis delays gut closure, weakens barrier integrity, and skews immunity toward pro-inflammatory responses. Feeding strategies such as colostrum timing, milk replacers, and the addition of starter fiber and probiotics can alter microbial communities within days. Nonetheless, challenges remain in standardizing neonatal feeding practices, identifying microbe metabolite indicators of gut health, and integrating precision feeding technologies. By mapping the three-way interaction among host, microbiota, and diet, this review offers a blueprint for neonatal ruminant feeding that enhances both animal welfare and the productivity of future ruminant systems. This review also provides a novel mechanistic integration of host genetics, gut microbiota succession, and dietary interventions in young ruminants, filling the gap of fragmented multi-omics analyses in existing literature and offering targeted insights for optimizing gastrointestinal development and production efficiency.
Aflatoxin B1 (AFB1), a potent mycotoxin, commonly contaminates feeds like maize and soybean, jeopardizing animal reproduction. Although AFB1 exposure is known to cause oxidative stress, immune activation, and cell death in oocytes and early embryos of several species, its effects on sheep remain unclear. This study aimed to investigate AFB1-induced damage in ovine oocytes and its underlying mechanisms. Quercetin (QT), a cost-effective flavonoid with antioxidant and anti-inflammatory properties, is potential to improve this damage. The mechanisms may involve non-canonical ferroptosis, an iron-dependent, lipid peroxidation-driven cell death pathway independent of canonical regulators GPX4 and TFR1. 4D Fast DIA-based micro-scale quantitative proteomics was conducted to identify the target proteins, and then immunofluorescence, qPCR, and parallel reaction monitoring (PRM) were conducted for the expression validation of the target proteins, and a series of analyses combined with the inhibitor experiments were conducted for the functional validation of the target proteins, including assessments of mitochondrial function (membrane potential ΔΨm, distribution, ATP levels), mitochondrial morphology observation by transmission electron microscopy (TEM), lipid peroxidation detection (LPO imaging, ROS and GSH detection), Fe2+ detection, endoplasmic reticulum staining, and early apoptosis signaling detection. OPA1 and ACSL4 are screened as the target proteins by micro-scale quantitative proteomics analysis, and immunofluorescence, qPCR and PRM validate their expression. Molecular docking reveals that there is an interaction of OPA1 and ACSL4, and QT exhibits stronger binding affinity to both OPA1 and ACSL4 than AFB1. AFB1 induces aberrant upregulation of OPA1 and ACSL4, disrupting mitochondrial cristae and membrane structure, impairing mitochondrial function and energy metabolism (including decreased mitochondrial membrane potential, abnormal distribution, and reduced ATP synthesis), promoting lipid peroxidation and Fe2+ accumulation, exacerbating endoplasmic reticulum stress and altering ROS/GSH levels, ultimately leading to ferroptosis in oocytes. Addition of QT ameliorates the AFB1-induced abnormal expression of OPA1 and ACSL4. QT alleviates AFB1-induced damage on ovine oocytes by suppressing non-canonical ferroptosis via the OPA1/ACSL4 pathway. These findings elucidate a novel mechanism underlying AFB1-mediated reproductive toxicity and provide a theoretical foundation for applying QT to mitigate AFB1-induced reproductive impairment and improve livestock health.
Reproductive efficiency in Nellore heifers is fundamental to the profitability and sustainability of beef production in tropical regions, where environmental stress can cause genotype-environment (G×E) interactions that affect fertility. Using 200,258 and 299,885 phenotypic records for heifer early pregnancy (HP) and heifer rebreeding (HR), respectively, we investigated the genetic basis of reproductive plasticity via single-step genomic reaction norms across a continuous environmental gradient (EG) defined from yearling weight records as a proxy for environmental quality. Genomic analyses included 22,556 animals (21,456 females and 1,100 sires) with genotypes imputed to 409,617 single-nucleotide polymorphisms (SNPs). We then performed genome-wide association analyses of the reaction norm intercept (genetic merit) and slope (environmental sensitivity), followed by multi-trait summary analyses and Bayesian fine-mapping of significant loci using imputed whole-genome sequence variants within ±100 kb windows around lead SNPs. Heritability for both traits increased with environmental quality, indicating environment-dependent expression of genetic variance. Genetic correlations for HP and HR across the environmental gradient ranged from 0.15 to 0.98, supporting substantial G×E interactions and reranking between low and high environmental conditions. Multi-trait analyses of reaction norm parameters identified 482 significant signals for the intercept and 700 for the slope. Intercept-associated loci were enriched for lipid metabolism, embryonic development, estrous regulation, and hypothalamic-pituitary signaling, whereas slope-associated loci highlighted endocrine signaling, metabolic plasticity, and neuroendocrine feedback responsive to contrasting post-weaning nutritional and management conditions captured by the EG. Fine-mapping refined associations to 146 (intercept) and 149 (slope) putative loci for HP, and 117 (intercept) and 167 (slope) for HR, supported by high posterior probabilities and Bayes factors. Candidate variants mapped to endocrine and metabolic regulators, including IGF1, LEP, GHRL, GNRHR, KISS1, MAPK3, PLAG1, INSR, and LHCGR. G × E interactions play a key role in shaping the genetic architecture of reproductive efficiency in Nellore heifers. Integrating reaction norm, multi-trait GWAS, and fine-mapping highlighted loci, affecting both genetic merit and environmental sensitivity of fertility, providing targets to select more resilient Nellore females for tropical systems.
Weaning piglets are highly susceptible to enterotoxigenic Escherichia coli (ETEC) infections, which can cause intestinal barrier function dysfunction and death. However, there is still a lack of efficient, economical, and safe nutritional interventions. This study aimed to investigate the effects of combining butyrate with niacin on intestinal barrier function repair and resistance to ETEC infection in weaned piglets. In this study, two 14-d animal experiments were designed to observe the optimal butyrate-to-niacin ratio and assess their responses to the ETEC challenge. Supplementation with butyrate and niacin at a ratio of 100:2 (2,000 mg/kg butyrate and 40 mg/kg niacin, BN2) increased the average daily gain (ADG) and reduced the diarrhea incidence. We also observed an increase in the levels of nicotinamide adenine dinucleotide (NAD) in the colon of weaned piglets. Notably, BN2 promoted amino acid anabolism in the colon and enhanced glycolysis and the tricarboxylic acid (TCA) cycle by increasing the acetylation of key enzymes in the TCA. Furthermore, BN2 enhanced the expression of indispensable genes for the colonic mucosal barrier, including antimicrobial peptides such as porcine β defensin 1 (pBD1), porcine β defensin 2 (pBD2), and proline-arginine rich 39-amino acid peptide (PR39), tight junction proteins, and improved colonic microbiome composition. Based on these findings, we found that BN2 alleviated growth restriction and diarrhea, and modulated the expression of antimicrobial peptides, tight junction proteins, and cytokines to reduce colonic barrier function dysfunction in weaned piglets challenged with ETEC. Mechanistically, we confirmed that BN2 elevated the protein expression of acetylation of histone 3 lysin 27 (H3K27ac) and enhanced the binding of acH3K27 to the promoter regions of pBD1 and PR39. Supplementation with BN2 improved growth performance, supported colonic barrier function repair, and enhanced disease resistance in weaned piglets challenged with ETEC. This offers new insights into nutritional strategies for intestinal barrier function repair of piglets infected with ETEC.
Under high-concentrate feeding conditions, ruminants often experience rumen microecological imbalance and dysfunction, which can impair growth performance and increase the risk of antibiotic resistance gene (ARG) dissemination. To evaluate the ameliorative effects of Aspergillus niger (A. niger) cultures, fattening sheep were randomly allocated into the following five groups: a control group (CON), a control diet supplemented with 250, 500, or 1,000 mg/kg A. niger cultures (designated as LA, MA, and HA, respectively); and an antibiotic group supplemented with 5,000 mg/kg chlortetracycline premix (AN). Microbial community analysis indicated that several bacterial taxa, including Succinivibrio sp900317105, Prevotella sp002353485, Quinella sp017515635, Quinella sp015206805, and Prevotella sp900320255, were significantly enriched in the A. niger culture-supplemented groups (P < 0.05). ARG profiling showed that the abundance of tetracycline resistance genes was significantly lower in all A. niger groups compared with the CON and AN groups (P < 0.05), while β-lactam resistance genes were significantly reduced in the HA group (P < 0.05). Furthermore, the abundances of Rank I and Rank II ARGs were significantly higher in the AN group than in the other groups, whereas the abundances of Rank II and Rank IV ARGs were significantly lower in the A. niger culture groups than in the CON and AN groups. Metabolomic analysis further demonstrated that supplementation with A. niger cultures significantly decreased the concentration of N-decanoyl-L-homoserine lactone (P < 0.05) while increasing the levels of N-3-oxotetradec-7Z-enoyl-L-homoserine lactone, indole-3-methyl acetate, and indole-3-propionic acid (P < 0.05). These findings suggest that A. niger cultures can reduce the abundance of ARGs and mitigate the risk of ARG dissemination by modulating the rumen microbial community and associated metabolites.
Late gestation represents an important developmental window for fetal skeletal muscle formation, which is associated with postnatal body growth, metabolic health and mobility in offspring. Although elevated circulating non-esterified fatty acids (NEFA) in dairy cows have been extensively studied in relation to milk production, their associations with fetal and neonatal skeletal muscle development remain unclear. Sixty healthy Holstein cows with similar body weight, parity and calving dates were enrolled in this study after dry-off, and retrospectively assigned to low NEFA (n = 30, 263 ± 8.8 μmol/L) and high NEFA group (n = 30, 379 ± 9.1 μmol/L) according to serum NEFA concentrations at 1, 3, 5, and 7 weeks after dry-off. Skeletal muscle was collected from calves at birth and at one month of age to assess the impacts on offspring muscle mass, fiber morphology and metabolic functions. C2C12 myoblasts were also used to assess the NEFA addition on myogenesis in vitro. Cows with high NEFA (within the physiological prepartum range) in the dry period had no effect on calf birth weight (P = 0.15), but exhibited reduced biceps femoris (P = 0.05) and semitendinosus muscle mass (P < 0.01). Moreover, calves in the high-NEFA group shifted the muscle fiber composition, characterized by a lower proportion of fast-twitch fibers (P < 0.01) and a higher proportion of slow-twitch fibers (P = 0.01). The protein abundance of mitochondrial fission-related markers dynamin-related protein 1 (DRP1) and mitochondrial fission protein 1 (FIS1), as well as stimulator of interferon genes (STING)-associated inflammatory markers, was increased in skeletal muscle tissue of calves born to high-NEFA cows (P < 0.01). These alterations were persistently observed at one month of age. In vitro, NEFA supplementation reduced myogenic differentiation and fast-twitch myofiber formation (P < 0.01), while increasing the expression of mitochondrial dynamics-related genes and inflammatory markers (P < 0.01). Overall, this study reveals that the calves born to mothers with elevated NEFA during the dry period are associated with altered skeletal muscle development and changes in the expression of mitochondrial dynamics- and inflammation-related markers.
The overuse of antibiotics in both veterinary and human medicine has resulted in the emergence of antibiotic-resistant bacteria, prompting a search for effective alternatives. Antimicrobial peptides (AMPs) are short, often cationic, peptide-based molecules with antimicrobial and immunomodulatory activity, which makes them promising alternatives to conventional antibiotics in poultry production. From a prior machine-learning-guided screen of 875 candidate AMPs against a wide bacterial panel, 62 exhibited activity against avian pathogenic Escherichia coli (APEC) and low in vitro hemolytic and cytotoxic activity. We selected three lead AMPs from this list (named TeRu4, TeBi1, and PeNi4), and evaluated their in vitro and in vivo efficacy, safety, and immunomodulatory potential for use in poultry farming. In animal experiments, AMPs were administered via in ovo injection on d 18 of embryonic development. In APEC challenge trials, yolk sacs were inoculated with APEC post-hatch to assess early chick mortality, while in pen trials, birds were raised in a commercial production setting for 35 d. For challenged birds, TeBi1 (10 μg/egg) significantly reduced culture-positive rates for APEC in the air sac and pericardium, increased body weight by 50% and reduced cytokine transcript levels by 10%-30% on d 7 post hatch. In HD11 chicken macrophage-like cultured cells, TeRu4 (16 μg/mL) suppressed lipopolysaccharide (LPS)-induced pro-inflammatory cytokine transcript levels. In pen trials, TeRu4 (20 μg/egg) increased the survival probability of female birds by 4.9%, while TeBi1 (20 μg/egg) increased the survival probability of all birds by 4.4%, by d 35. Gene expression analysis revealed AMP- and sex-specific cytokine responses. In pen trials, no significant differences were observed in mean weights, feed conversion ratio (FCR), and flock uniformity on d 35. By integrating high-throughput in ovo automation with large-scale commercial pen trials, this study provides a systematic translational bridge from in silico AI discovery to field-relevant poultry production interventions. These findings demonstrate that TeBi1 and TeRu4 are promising antibiotic alternatives that improve survival, modulate immune responses, and maintain normal growth performance in broiler chickens in this experimental setting.
Intestinal epithelial cells are supported by dynamic cellular heterogeneity, which is critical for maintaining intestinal homeostasis. Recently, intestinal organoid models have gained attention as in vitro platforms because they can recapitulate the structural, functional, and cellular complexity of the small intestine. In this study, we developed porcine intestinal organoids and investigated time-dependent transcriptomic changes using single-cell RNA sequencing. Furthermore, to assess their applicability as an in vitro toxicity model, the organoids were exposed to the mycotoxins deoxynivalenol and zearalenone. The established organoids exhibited stable long-term culture up to passage 10 (32 d) and showed high genetic similarity to native small intestinal tissue across three regions: the duodenum, jejunum, and ileum. In various intestinal epithelial cell types, including transit-amplifying cells, enteroendocrine cells, goblet cells, Paneth cells, and other epithelial cell types, were identified in the organoids. Single-cell RNA sequencing classified the organoids into multiple distinct cell populations, including stem cells, transit-amplifying cells, secretory progenitors, enterocytes, enteroendocrine cells, goblet cells, and Paneth cells, demonstrating dynamic cellular heterogeneity. The organoids also recapitulated key intestinal functions, such as nutrient absorption and epithelial barrier formation, similar to those of the native small intestinal epithelium. Under these conditions, the cytotoxic effects of deoxynivalenol and zearalenone were evaluated. Treatment with these mycotoxins resulted in decreased cell viability, impaired intestinal barrier function, and altered rates of proliferation and differentiation, including those of enteroendocrine, goblet, and Paneth cell populations. This study provides fundamental insights into the growth and differentiation of small intestinal epithelial cells by analyzing timeline-specific organoids using single-cell sequencing. Additionally, it evaluates the toxicity of mycotoxins under conditions that closely resemble those of the small intestine, providing more physiologically relevant data than existing in vitro models and serving as a reliable toxicity assessment model.