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Evaluation of circulating leukocyte transcriptome and its relationship with immune function and blood markers in dairy cows during the transition period


Dairy cows during the transition period are faced with important physiological changes which include a dysfunctional immune system and an increased inflammatory state. New data are necessary to understand the key factors involved in the immune system regulation. Six dairy cows were sampled during transition period to investigate the leukocyte transcriptome changes and its relationship with blood biomarkers. Blood samples were collected at − 20 ± 2, − 3 ± 1, 3, and 7 days from parturition (DFP). Leukocyte transcriptome was analyzed by deep sequencing technology (Hiseq1000 Illumina, USA). Plasma was analyzed for metabolic biomarkers. Differentially expressed genes (DEG) were used to run an enrichment analysis through the Dynamic Impact Approach (DIA). Considering − 20 DFP as references time, the main KEGG impacted pathways were activated before calving (− 3 DFP) and were connected to lipid metabolism, lipid transport in plasma, and phagosome. The greatest differences were found after parturition with 281 DEG (179 upregulated and 102 downregulated). The activated pathways were mainly related to immunity and endocrine aspects, while metabolic pathways related to lipid and amino acid metabolism were inhibited. Plasma BHBA had a substantial inhibitory impact on KEGG pathways related to DNA replication and cell cycle, while plasma IL-1β had an inhibitory impact on fatty acid elongation in mitochondria and an activated impact in several pathways related to cellular energy metabolism. Overall, this study confirmed that many changes in lipid metabolism and immune competence of the circulating leukocytes occurred in dairy cow around calving. Interestingly, BHBA and IL-1β connected with the transcriptome.

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Availability of data and material

The datasets generated and analyzed during the current study are available from the corresponding author on reasonable request. The DIA software is not publicly available but can be requested from the developer. KEGG databases are free and available online.



alkaline phosphatase


body condition score




differentially expressed genes


days from parturition


Dynamic Impact Approach


γ-glutamil transferase


aspartate aminotransferase


Kyoto Encyclopedia of Genes and Genomes




negative energy balance


non-esterified fatty acids

NO2 :


NO3 :


NOx :

total nitric oxide metabolites




total reactive oxygen metabolites.


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The authors gratefully acknowledge Dott.ssa Annarita Ferrari for her support during biochemical chemistry analysis and Prof. Giuseppe Bertoni (Department of Animal Sciences, Food and Nutrition, Università Cattolica del Sacro Cuore, Piacenza, Italy) for his help and suggestions in result interpretation


This study was funded by the “Romeo ed Enrica Invernizzi foundation,” Milan, Italy.

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Correspondence to Andrea Minuti.

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Minuti, A., Jahan, N., Lopreiato, V. et al. Evaluation of circulating leukocyte transcriptome and its relationship with immune function and blood markers in dairy cows during the transition period. Funct Integr Genomics 20, 293–305 (2020). https://doi.org/10.1007/s10142-019-00720-0

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  • Immune system
  • Transition period
  • Dairy cows
  • Transcriptomics
  • Plasma metabolites
  • RNA-Seq