ABSTRACT Emaciation syndrome presents a major challenge to aquaculture by reducing growth performance and survival in cultured fish. However, its underlying physiological and molecular mechanisms remain poorly understood. This study investigated the pathological characteristics and transcriptomic alterations associated with severe emaciation in the leopard coral grouper ( Plectropomus leopardus ). Histological examination revealed extensive intestinal villus atrophy, hepatic lipid depletion, and pronounced skeletal muscle fiber shrinkage in emaciated fish. RNA‐seq analysis across the intestine, liver, and muscle identified 3327, 4482, and 5496 differentially expressed genes (DEGs), respectively, between the emaciated and control groups. Functional enrichment analysis indicated that suppression of digestive enzyme genes (e.g., cela1 , cela3b , and cpa5 ) in the intestine, activation of lipid catabolism and gluconeogenic pathways (including pck1 ) in the liver, and upregulation of proteasome subunit genes ( psma1 , psmb1 , and psmc3 ) in the muscle were key molecular alterations associated with emaciation syndrome. Quantitative reverse transcription polymerase chain reaction (qRT‐PCR) and in situ hybridization (ISH) validated the tissue‐specific transcriptional patterns of these representative DEGs. Collectively, these findings suggest coordinated tissue‐specific alterations involving intestinal digestive and absorptive function, hepatic metabolic regulation, and muscle protein turnover in emaciated groupers. This integrative analysis provides new insights into the molecular responses of the intestine, liver, and muscle to emaciation and offers a useful molecular reference for future studies on early diagnosis and health‐management strategies in aquaculture.
ABSTRACT Dietary fiber (DF), long recognized for its contributions to digestive health and satiety regulation, is now acknowledged as a crucial modulator of meat quality in food‐producing animals through its significant impact on the gut‐muscle axis. In this brief review, we synthesize recent research findings, focusing on the mechanisms by which DF influences key meat quality attributes, including tenderness, juiciness, color, flavor, and nutritional profile. Specifically, these mechanisms comprise the regulation of gut microbiota composition, modulation of metabolites produced via fermentation pathways (e.g., short‐chain fatty acids, SCFAs), enhancement of intestinal barrier function, mitigation of systemic inflammation, and optimization of nutrient partitioning. We further explain how different types of DF, characterized by solubility, source, and structure, elicit distinct microbial responses and metabolic effects that influence muscle fiber composition, metabolic activity, antioxidant capacity, and post‐mortem biochemical processes. The review emphasizes a shift in perspective, transitioning from viewing DF as a simple bulking agent to recognizing it as a prebiotic that regulates host‐microbe crosstalk and exerts direct effects on muscle physiology. Comprehending these intricate interactions provides sustainable methodologies for improving meat quality through customized DF interventions, moving beyond conventional nutritional strategies. Additionally, this review addresses critical challenges, such as the optimization of DF inclusion levels, species‐specific effects of supplementation, and future research directions in the field.
ABSTRACT Porcine epidemic diarrhea virus (PEDV) is an enteric coronavirus that causes acute gastrointestinal infection in neonatal piglets, characterized by severe diarrhea, high transmissibility, and elevated mortality rates. As a major threat to the global swine industry, PEDV outbreaks incur substantial economic losses in pork production. After extensive circulation over prolonged periods, PEDV has accumulated a series of mutations, resulting in strains with enhanced transmissibility and pathogenicity. Currently, available vaccines provide insufficient protection against PEDV, and no commercial antiviral drugs or PEDV‐resistant pig breeds are available for clinical prevention and treatment. This review summarizes the structural and functional characteristics of PEDV, as well as the current understanding of antiviral compounds, host factors, and virus–host interactions during PEDV infection. Gene‐editing technologies have been successfully applied to generate antiviral pigs, offering valuable insights for the development of PEDV‐resistant pigs. Collectively, this review provides critical information for deciphering the PEDV infection process and associated host factors and facilitates the prevention and control of PEDV infection.
ABSTRACT Milk oligosaccharides (MOs) are an abundant, diverse, and complex class of carbohydrates found in breast milk that are increasingly recognized as key determinants of intestinal health during early life. Their composition and structural diversity differ markedly across mammalian species, shaping host–microbe interactions and contributing to intestinal barrier homeostasis. This review systematically summarizes the compositional and structural variations of MOs across mammals and discusses their regulatory roles in intestinal homeostasis within a structure‐based classification framework. These roles are mediated through their coordinated effects on gut microbiota composition and function, the intestinal epithelial barrier, and immune function. By integrating these dimensions, this review outlines a conceptual framework for MO‐host interactions and highlights the potential of MO‐derived nutritional strategies to support mammalian intestinal health in early life.
ABSTRACT Fungal infections, including saprolegniasis and opportunistic mycoses, pose persistent challenges in aquaculture, driving the search for effective and safe antifungal agents. Clotrimazole (CLO), a clinically established imidazole, exhibits broad‐spectrum antifungal activity by inhibiting sterol 14α‐demethylase, disrupting ergosterol biosynthesis, and compromising fungal cell membrane integrity. This review synthesizes current knowledge on CLO's potential role in fungal disease, particularly saprolegniasis management, highlighting mechanistic plausibility alongside significant limitations largely based on in vitro and mechanistic data. Key challenges include the paucity of replicated in vivo efficacy and toxicokinetic data in representative aquaculture species, unresolved residue and food‐safety considerations, regulatory constraints, and potential environmental impacts, including selection for antifungal resistance. This review examines pharmacokinetic and pharmacodynamic considerations, environmental fate, and ecotoxicological risks, emphasizing the need for closed‐system testing and targeted delivery to mitigate off‐target exposure. Future research directions focus on species‐specific in vivo trials, good laboratory practice‐compliant residue and tissue distribution studies, advanced formulation strategies such as mucoadhesive carriers and nanoparticle delivery, and integrated One Health‐oriented stewardship frameworks, including resistance monitoring. By framing CLO as a conditional candidate for controlled aquaculture use, this review provides a comprehensive blueprint for rational evaluation, highlighting the balance between therapeutic promise and precautionary risk management in aquatic veterinary pharmacology, and offers a scientific basis for future research priorities and informed policy decisions.
ABSTRACT Nipah virus (NiV) is a zoonotic paramyxovirus classified as a WHO priority pathogen, which causes severe respiratory disease and encephalitis in humans with high mortality. Currently, no licensed vaccines or specific antiviral therapies are available. A major obstacle in NiV research is the lack of immunocompetent rodent models that endogenously express human‐compatible entry receptors Ephrin‐B2 (EFNB2) and Ephrin‐B3 (EFNB3). Here, we report the generation and characterization of two novel knock‐in mouse models using the CRISPR/Cas9 system. The human EFNB2 or EFNB3 gene, driven by the human keratin 18 (K18) promoter, was site‐specifically integrated into the mouse Hipp11 ( H11 ) safe‐harbor locus. Founder mice exhibited stable germline transmission without obvious developmental abnormalities. These established models provide a physiologically relevant and genetically engineered platform for studying NiV entry mechanisms, pathogenesis, and the development of vaccines and antiviral therapeutics.
This study was conducted to evaluate the effects of an organic acid blend on growth performance, redox homeostasis, and intestinal microbiota of broilers raised in an antibiotic-free system during the winter season in China, with low ambient temperatures and reduced ventilation. A total of 1080 1-day-old male broilers were randomly assigned to one of three treatment groups, with 18 replicates of 20 broilers each. All broilers were fed the same basal diet and provided either water (control, CON), water with antibiotics (ABx), or water with an organic acid blend (OA) for 5 weeks. The results showed that both ABx (+2.69-2.75%) and OA (+2.95-3.98%) treatments significantly increased body weight gain from Days 1 to 35 and body weight on Day 35 compared to the CON group. OA treatment also upregulated the mRNA expression of key immune-related markers in the jejunum, including cytokines and transcription factors (IL-1 beta and NF-kappa B), tight junction proteins (ZO-1 and occludin), and host defense peptides (AvBD-1 and cathelicidin-1). Furthermore, OA treatment significantly reduced serum LPS concentrations (p < 0.05). Microbial analysis using principal coordinates analysis (PCoA) revealed significant differentiation (p < 0.05) between the CON and the ABx or OA groups. LEfSe analysis identified 22 discriminative microbial features across the groups. Both ABx and OA treatments significantly increased the abundance of Intestinimonas, Bryobacter, and Rhodanobacter compared to the CON group. Additionally, ABx treatment significantly increased Lachnospiraceae FCS020, whereas OA treatment significantly increased Lachnospiraceae NK4A136 compared to ABx, with no significant difference observed compared to the CON group. In conclusion, the organic acid blend positively influenced broiler growth performance and microbial composition, suggesting its potential as a viable alternative to antibiotics for promoting growth and health in antibiotic-free production systems.
Riboflavin is a crucial micronutrient essential for maintaining cellular homeostasis, acting as an important precursor for flavoproteins that utilize flavin mononucleotide and flavin adenine dinucleotide as cofactors. Some flavoproteins play roles in the folding of newly synthesized proteins within the endoplasmic reticulum. However, the specific proteins whose folding is influenced by riboflavin remain unidentified. Disulfide bond-modified proteomics to identify proteins whose folding is influenced by riboflavin. Our findings identified 34 proteins in HepG2 cells, a highly riboflavin-dependent model, with riboflavin-dependent disulfide bond modifications. These proteins are primarily involved in metabolic pathways such as endoplasmic reticulum protein processing, antigen processing and presentation, glycolysis/gluconeogenesis, lysosomal function, and amino acid biosynthesis. Furthermore, the accumulation of these unfolded or misfolded proteins in the endoplasmic reticulum enhanced the activity of the CHOP promoter, leading to increased expression of the CHOP gene and protein, thereby triggering the cellular apoptosis pathway. For the first time, we have identified riboflavin-dependent protein folding substrates using disulfide bond-modified proteomics. These findings may provide new perspectives for future therapies targeting endoplasmic reticulum stress-related diseases.
Ostreid herpesvirus 1 (OsHV-1) poses a major threat to bivalve aquaculture, but its impact on blood clams (Anadara broughtonii) remains poorly understood. This study integrated epidemiological and genomic data from 72 batches (1358 samples) collected across Chinese production systems (2019-2025). OsHV-1 was detected in hatcheries, holding facilities, and wild populations, with high viral loads linked to mass mortalities. Prevalence was significantly higher in land-based facilities (72%-74%) than in open-water systems (0%-50%; p < 0.001). Asymptomatic carriers, including wild populations and clams imported from the Republic of Korea, harbored lower viral loads than diseased individuals, acting as cryptic reservoirs. Temperature analysis identified disease onset at 13 degrees C and a viral load peak at 18.9 degrees C (95% CI: 16.4 degrees C-22.3 degrees C), approximately 3 degrees C-5 degrees C lower than those reported for oysters, reflecting host-specific viral adaptation. Effluent from land-based facilities tested virus-positive, indicating a potential transmission route. Random forest analysis identified source type as the strongest predictor of infection (AUC = 0.911). Mortality risk was strongly associated with temperature (OR = 2.65, p = 0.017) and viral load (OR = 4.23, p = 0.006). Genomic analysis revealed a distinct, host-specific lineage with no clear temporal progression. These findings demonstrate that effluent discharge, asymptomatic wild carriers, and international trade sustain OsHV-1 endemicity in blood clam aquaculture, highlighting the need for enhanced biosecurity measures calibrated to this cold-adapted pathosystem.
Ketotic cows undergo severe negative energy balance, in which increased circulating non-esterified fatty acids, specifically palmitic acid (PA), serve as primary drivers of liver injury. Yet, effective strategies for alleviating this lipotoxic injury remain limited. Although tryptophan metabolite indole-3-acetic acid (3-IAA) has been documented to exert hepatoprotective effects in non-ruminants, its role in alleviating lipotoxicity-induced injury in bovine hepatocytes remains unclear. In the present study, ketotic cows exhibited increased serum beta-hydroxybutyrate and non-esterified fatty acid concentrations and hepatic triacylglycerol accumulation and decreased serum glucose and 3-IAA concentrations. Notably, serum 3-IAA concentration negatively correlated with the hepatic triacylglycerol content. In primary bovine hepatocytes, treatment with 200 mu mol/L 3-IAA improved viability and reduced cytotoxicity under PA challenge. Transcriptomic analysis further indicated that 3-IAA mainly regulated pathways related to lipid metabolism and endoplasmic reticulum (ER) stress. Further analysis showed that 3-IAA attenuated PA-induced lipid accumulation and downregulated sterol regulatory element-binding protein 1c protein abundance and its downstream target gene expression, while upregulating the expression of fatty acid oxidation-related molecules, indicating reduced lipogenesis and enhanced fatty acid oxidation. In addition, 3-IAA markedly alleviated PA-induced ER stress, as evidenced by reduced expression of ER stress-related markers. Notably, 3-IAA increased sirtuin 3 (SIRT3) protein and mRNA expression in PA-treated hepatocytes, whereas SIRT3 knockdown partially abrogated the protective effects of 3-IAA. Collectively, these findings demonstrate that 3-IAA mitigates PA-induced lipid accumulation and ER stress in bovine hepatocytes, at least in part through a SIRT3-dependent mechanism, highlighting its potential as an endogenous regulator of hepatic homeostasis in ketotic cows.
Avian salmonellosis, caused by Salmonella species, represents a significant zoonotic disease that substantially impacts both the poultry industry and public health security. Although domestic poultry breeds exhibit distinct interbreed variations in resistance to Salmonella typhimurium (ST) infection, the underlying molecular mechanisms remain poorly understood. In this study, we employed integrated immunophenotypic (including H/L ratio, IFN-gamma, IL-1 beta, and IL-8) and splenic transcriptomic analyses to identify candidate genes associated with differential resistance to Salmonella infection across chicken breeds. To this end, 28-day-old Guang Ming No. 2 broiler line B (GM) and Beijing-You (BY) chickens were challenged with Salmonella typhimurium (ST) at a dose of 2.5 & times; 1010 CFU/mL. Through an integrated approach combining differential expression analysis and WGCNA, we identified three key candidate genes-EGR1, FOS, and DUSP1-associated with differential resistance to Salmonella infection between chicken breeds. Functional validation confirmed that FOS and DUSP1 play significant roles in modulating immune cell responses to ST infection. Furthermore, we uncovered a novel regulatory mechanism whereby EGR1 negatively regulates DUSP1. These findings provide potential candidate targets for poultry disease resistance research.
Skeletal muscle development not only determines the growth performance and meat quality of livestock, but is also closely associated with human metabolic health. ZBED6 has been identified as a transcriptional repressor of IGF2, yet its role and molecular mechanism in skeletal muscle development remain unclear. In this study, we employed a ZBED6 knockout (KO) Bama pig model and found that loss of ZBED6 led to increased muscle mass and muscle fiber hypertrophy, suggesting that ZBED6 plays an inhibitory role in muscle growth. To further elucidate the underlying mechanisms, we performed integrated transcriptomic and functional analyses to investigate the ZBED6-CDKN1A axis. RNA sequencing identified differentially expressed genes across seven tissues from 8-month-old KO and wild-type pigs, and WGCNA revealed a skeletal muscle-specific module enriched in myogenic pathways. By integrating this module with transcriptome data from longissimus dorsi at 5 and 8 months of age and ChIP-seq data, CDKN1A was identified as a central hub gene. Dual-luciferase reporter assays demonstrated that ZBED6 binds to a conserved GCTCG motif in the CDKN1A promoters of pigs and mice to repress its transcription. Consistently, in C2C12 cells, ZBED6 knockdown upregulated the expression of myogenic markers (MyoD, MyoG, and MyHC) and promoted myotube formation, whereas silencing CDKN1A attenuated this effect. Conversely, ZBED6 overexpression reduced CDKN1A expression and suppressed differentiation, confirming its bidirectional regulatory role. Collectively, our data identify CDKN1A as a transcriptional target of ZBED6 and show that ZBED6 constrains skeletal muscle development by repressing CDKN1A.
ABSTRACT Plant‐based beverages that attempt to mimic milk are increasingly popular with consumers, but there are profound differences in nutritional composition and microstructure compared to milk. A structural nutrition approach is needed to understand the effect on digestibility of microstructures, bioaccessibility and bioavailability of components, and ultimately on nutritional delivery and human health.
ABSTRACT Dairy cattle's lactation efficiency has increased over the last decades, which is mainly attributed to high milk production and elevated nutrient requirements, especially protein and amino acids (AA) requirements. The transition period is considered one of the most crucial periods of a dairy cow's life, with the largest number of health disorders happening within the first 14 days in milk. Changes in dairy cow metabolic conditions might have had a severe impact on dairy cow health, fertility, and production, particularly milk and milk component yields. One of the applied strategies to prevent these negative impacts is balancing amino acids profile in dairy cattle feeds, primarily, rumen bypass methionine and lysine (RPM and RPL) to meet the requirements through increasing the metabolizable amino acid (AA) for intestinal absorption. NASEM (2021) presented a novel concept based on considering 5 indispensable amino acid (IAA) rather than the more aggregated metabolizable protein (MP) and the Lys–Met ratio. Continued attempts to improve the understanding of AA pathways make the possibility of new strategies to minimize the negative impacts on dairy cow metabolic status changes. Previous research observed beneficial impacts of supplementing RPM and RPL in transition dairy cow diets either/both (pre‐and postpartum periods) on dry matter intake, milk performance, plasma AA concentration, the occurrence rate of metabolic disorders, and immune response; however, the findings were inconsistent. Therefore, this review focused on the inconsistencies in the data and sought research gaps in this area.
ABSTRACT Sustainable dairy development is crucial for global food security, as it converts feed resources into nutrient‐dense human food while supporting soil fertility through manure recycling. Milk provides high‐quality protein and essential micronutrients, yet rising production demands have intensified challenges related to animal welfare and environmental impacts. Addressing these issues requires integrated strategies, including precision monitoring technologies, enteric methane inhibitors, and improved manure management. A balanced approach aligning productivity with animal health and ecological integrity is essential for future dairy systems.
ABSTRACT Circulating concentrations of nonesterified fatty acids (NEFAs) are elevated due to lipid mobilization from adipose tissue in periparturient dairy cows. Although this metabolic adaptation facilitates energy homeostasis under the negative energy balance condition, sustained systemic NEFA overload induces profound hepatic impairment. Emerging evidence identifies excessive NEFAs to be the pathophysiological cornerstone of periparturient disorders; however, the precise molecular mechanisms underlying NEFA‐induced hepatotoxicity remain incompletely characterized, hindering the development of effective preventive and therapeutic strategies. This literature review synthesizes contemporary insights into key cellular pathways implicated in NEFA‐mediated hepatotoxicity: disorders in lipid and carbohydrate metabolism, impairment of autophagy, excessive inflammatory response, mitochondrial dysfunction, oxidative stress, endoplasmic reticulum stress, and finally, cell death. Critical analysis reveals two underexplored dimensions in current research paradigms: (1) The dynamic composition of circulating NEFAs modulates hepatotoxic potency through differential membrane incorporation and signaling pathway activation, suggesting that improving blood NEFA composition through dietary fat supplementation offers a potential strategy; and (2) the periparturient inflammatory milieu potentiates NEFA toxicity, suggesting targeted anti‐inflammatory interventions ameliorate transition period adaptation. Consequently, this review advances our mechanistic understanding while providing translational frameworks for improving periparturient management through precision nutrition and therapeutic development.
Depressive disorders are associated with accelerated tumor progression and poor clinical outcomes. Accumulating evidence indicates that psychiatric conditions and tumor development are tightly interconnected, yet the mechanisms by which depression remodels the tumor immune microenvironment remain poorly defined. To investigate how chronic stress influences tumor progression and host immunity, we established murine models of depression-like behavior and depression-tumor multimorbidity. Depression-like phenotypes were validated through behavioral tests and hormone measurements, followed by comprehensive analyses of tumor growth and immune landscapes using transcriptomic sequencing, bioinformatic analyses, flow cytometry, and immunohistochemistry. Our results demonstrated that depressive states significantly promoted tumor growth. Transcriptomic profiling revealed widespread downregulation of multiple immune-related pathways in depression-associated tumors. Immunophenotyping further showed that depression markedly remodeled the tumor immune microenvironment, characterized by reduced intratumoral CD8+ T cell abundance, enhanced functional exhaustion, and increased PD-1 expression, collectively contributing to impaired antitumor immunity. These findings indicated that depression accelerates tumor progression in parallel with the disruption of immune homeostasis and induction of CD8+ T cell dysfunction. This study underscores the importance of systemic crosstalk between behavioral states and tumor immunity in the context of depression-tumor multimorbidity and provides a theoretical basis for integrating psychosocial interventions into cancer therapy. Furthermore, this work highlights the correlation between psychological state and tumor progression accompanied by systemic immune modulation, supporting the broader One Health perspective on the interconnection between physical and psychological multimorbidity.
Horns are emblematic ruminant cranial appendages, yet their molecular basis and phenotypic diversity remain incompletely understood. Here we built a systematic framework for sheep horn biology by integrating multi-tissue transcriptomics, cross-species conservation, fine-grained tissue comparisons, horn-type contrasts, and allele-specific expression (ASE). We first profiled 160 RNA-seq samples across 16 tissues and identified 1608 horn tissue-specific genes, revealing horn-distinct programs strongly enriched in keratinization and epidermal differentiation (e.g., LORICRIN and KRT2). Next, cross-species analyses (sheep, goat, deer, and cattle) defined conserved horn-associated gene sets that couple epidermal modules with osteogenic components (e.g., COL1A1 and SOX9), supporting a composite epidermal-skeletal identity. We then compared horn with skin and periosteum to partition shared versus horn-specific programs, highlighting both keratinocyte-related regulators shared with skin and bone-matrix signals linked to periosteum, alongside horn-specific metabolic remodeling signatures. Furthermore, differential expression between large-horned (SHE) and scurred (SCU) individuals identified candidate regulators of horn size variation, including remodeling factors and transcriptional regulators (e.g., MMP9 and SNAI1). Finally, ASE analysis uncovered cis-regulated candidate drivers, including horn-enriched genes and a horn/skin-specific enhancer-linked variant at ABHD5. Collectively, our study defines a coherent transcriptional blueprint for sheep horn development and provides prioritized genes and regulatory loci underlying horn identity and size diversity.
