Transportation is frequently employed as required travel practice for laboratory animals today.Transportation, however, is a type of stressor known to influence physiological parameters and wellbeing of animals.To obtain a better understanding of molecular mechanisms associated with transport stress, pre-and post-transport serum and urine samples were collected from cynomolgus macaques to look for potential differences and metabolomic biomarkers.Proton nuclear magnetic resonance spectroscopy (1H-NMR)-base metabolomics strategy with orthogonal projection to latent structure with discriminant analysis (OPLS-DA) method was used to analize the biospecimen.Samples of serum and urine at pre-transport, arrival and 5 weeks subsequent were collected.Thirty-eight and 26 metabolites in urine and serum, respectively, were unambiguously assigned.Increased level of lactate, alanine, glutamate, glutamine, pyruvate, myo-Inositol, creatinine and urea, and decreased level of isoleucine and proline (P < 0.05) in serum were observed post-transport.Metabolic trajectories indicted the greatest changes were noted between pre-transport and on arrival.In conclusion, our investigations showed significant metabolic differences in serum and urine in different time point in cynomulgus monkeys after combined air and ground transport.These differences included altered energy metabolisms, amino acid metabolisms, lipid metabolisms and perturbations in gut microbiota.Moreover, NMR-based metabolomics approach is a promising tool in order to obtain insight into physiological status and to explore stress to affect health and welfare.