Heat stress poses serious threats to human and animal health by inducing systemic inflammation, oxidative stress, and intestinal barrier damage, yet the potential of functional food components in mitigating heat stress-associated health impairments remains insufficiently explored. This study used a chronic heat stress model in C57BL/6 J mice to compare the protective effects of β-glucan (BG) and mannan oligosaccharides (MOS) against heat stress-induced injury. The underlying mechanisms of each supplement were also systematically investigated. The results demonstrated that both BG and MOS effectively attenuated heat stress-induced body weight loss, elevated liver index, and systemic inflammatory responses, significantly reduced serum levels of interleukin-1β (IL-1β), interleukin-6 (IL-6), tumor necrosis factor- α (TNF-α), and heat shock protein 70 (HSP70), and restored antioxidant enzyme activity. Notably, BG exhibited superior efficacy in suppressing pro-inflammatory cytokines and restoring serum immunoglobulin A (IgA) levels. Regarding intestinal barrier integrity, both oligosaccharides markedly upregulated the colonic expression of tight junction proteins zonula occludens-1 (ZO-1), Claudin-1, and Occludin, decreased serum diamine oxidase (DAO) and lipopolysaccharide (LPS) levels, and partially alleviated heat stress-induced intestinal barrier disruption. Hepatic tissue analysis revealed that both BG and MOS ameliorated heat stress-induced hepatic inflammation and lipid metabolism dysfunction. This was achieved by suppressing TLR4 and iNOS expression while restoring the balance of CD36 and PPARα expression. Furthermore, fecal microbial diversity analysis revealed that MOS was associated with increased abundance of Lachnospiraceae-related taxa, which are known to include short-chain fatty acid-producing bacteria. These microbial changes may contribute to the maintenance of gut microecological homeostasis via distinct microbiota-associated pathways. Collectively, these findings suggest that BG and MOS may alleviate multi-level heat stress-induced damage, potentially in association with improved intestinal barrier-related markers, altered gut microbiota composition, and modulation of gut–liver axis-related responses, thereby providing preliminary evidence for their potential application as functional food components to address heat stress-related health challenges.

Background: Long-term high-fat diet (HFD) consumption induces hepatic lipid accumulation and metabolic dysregulation, but its integrated effects on the hepatic proteome and phosphoproteome remain incompletely understood. This study investigated HFD-induced hepatic molecular alterations using quantitative proteomics and phosphoproteomics. Methods: Eight-week-old male C57BL/6J mice were fed a control diet (CON) or HFD for 16 weeks, followed by phenotypic assessment, serum redox assays, and hepatic proteomic and phosphoproteomic analyses. Results: HFD-fed mice exhibited increased body weight, fat mass, liver weight, and marked hepatic lipid accumulation. Blood glucose levels and total pyruvate tolerance test area under the curve (AUC) were increased, whereas baseline-adjusted incremental AUC did not differ significantly. Serum malondialdehyde levels were elevated, whereas glutathione peroxidase activity was reduced, indicating impaired glucose homeostasis and redox balance. Proteomic analysis identified 15 differentially expressed proteins (4 upregulated and 11 downregulated), associated with lipid metabolism, biological oxidation, and xenobiotic metabolism. Gene set enrichment analysis identified significant positive enrichment of Class I peroxisomal membrane protein import and significant negative enrichment of cholesterol biosynthesis, gluconeogenesis, and xenobiotics. Phosphoproteomic analysis identified 8593 phosphoproteins and 28,180 phosphorylation sites, including 2045 differentially phosphorylated sites (1044 increased and 1001 decreased). Enrichment and exploratory network analyses implicated mitophagy, glucose metabolism, mTOR signaling, autophagy, oxidative phosphorylation, and cytoskeletal organization. Conclusions: Long-term HFD feeding induced obesity, glucose dysregulation, hepatic steatosis, and redox imbalance. Fatty acid oxidation and peroxisomal abundance patterns may reflect metabolic adaptation to lipid loading, while phosphosite abundance changes link nutrient sensing and autophagy regulation to the hepatic response.

This preprint has been withdrawn at the request of the authors, as they have identified major flaws that cannot be adequately resolved by releasing an updated version.

Heat stress, particularly during the prepartum period, compromises dairy cow health and has lasting effects on offspring. This study aimed to investigate the effects of maternal yeast culture (YC) supplementation during heat stress on the intestinal health of newborn calves, with a specific focus on elucidating the associated changes in jejunal morphology, microbial community, and metabolic profiles. Cows receiving YC had lower rectal temperatures, confirming mitigated heat stress. Their calves showed enhanced immune status (higher IgA, IgG) and reduced oxidative stress and metabolic markers. Jejunal structure and barrier integrity were improved. While microbial diversity was similar, specific bacterial abundances differed (e.g., Enterococcus spp. enriched in YC calves). Metabolomic analysis revealed modulated bile acid pathways and metabolites in YC calves, correlating with the microbial shifts. In conclusion, maternal YC supplementation during heat stress supports calf early-life gut health and development through maternal-offspring programming, optimizing gut microbiota, metabolism, and immunity.