摘要Heat stress(HS)damages livestock by adversely affecting physiological and immunological functions.However,fundamental understanding of the metabolic and immunological mechanisms in animals under HS remains elusive,particularly in steers.To understand the changes on metabolic and immune responses in steers under HS condition,we performed RNA-sequencing and proton nuclear magnetic resonance spectroscopy-based metabolomics on HS-free(temperature humidity index[THI]value:64.92±0.56)and HS-exposed(THI value:79.13±0.56)Jersey steer(n=8,body weight:559.67±32.72 kg).This study clarifies the metabolic changes in 3 biofluids(rumen fluid,serum,and urine)and the immune responses observed in the peripheral blood mononuclear cells of HS-exposed steers.This integrated approach allowed the discovery of HS-sensitive metabolic and immunological pathways.The metabolomic analysis indicated that HS-exposed steers showed potential HS biomarkers such as isocitrate,formate,creatine,and riboflavin(P＜0.05).Among them,there were several inte-grative metabolic pathways between rumen fluid and serum.Furthermore,HS altered mRNA expression and immune-related signaling pathways.A meta-analysis revealed that HS decreased riboflavin meta-bolism and the expression of glyoxylate and dicarboxylate metabolism-related genes.Moreover,meta-bolic pathways,such as the hypoxia-inducible factor-1 signaling pathway,were downregulated in immune cells by HS(P＜0.05).These findings,along with the datasets of pathways and phenotypic differences as potential biomarkers in steers,can support more in-depth research to elucidate the inter-related metabolic and immunological pathways.This would help suggest new strategies to ameliorate the effects of HS,including disease susceptibility and metabolic disorders,in Jersey steers.