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The transition period is a critical time for dairy cows as the animal is subjected to the physiological stress accompanying parturition. Immunosuppression and health status were examined during this period in 80 Holstein cows. Blood samples were taken from each cow 3, 2 and 1 week before and after calving, and at calving (0 day). RNA was extracted and subjected to real‐time PCR to determine mRNA levels for the immune‐related genes TLR 2, 4, 6, 7 and β‐defensin 5 in addition to the reproduction‐related genes prolactin and IGF‐I. Results showed significant up‐regulation of pro‐inflammatory‐selected genes, TLR 4, 6 7 and β‐defensin 5 at the third‐week post‐calving; however, earlier periods had lower expression of such genes. In contrast, the immunosuppression biomarker TLR2 gene was up‐regulated at calving and 1 week after parturition and then down‐regulated again at second and third …