Abstract
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.
Similar content being viewed by others
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.
Abbreviations
- ALP:
-
alkaline phosphatase
- BCS:
-
body condition score
- BHBA:
-
β-hydroxybutyrate
- DEG:
-
differentially expressed genes
- DFP:
-
days from parturition
- DIA:
-
Dynamic Impact Approach
- GGT:
-
γ-glutamil transferase
- GOT:
-
aspartate aminotransferase
- KEGG:
-
Kyoto Encyclopedia of Genes and Genomes
- LPS:
-
lipopolysaccharide
- NEB:
-
negative energy balance
- NEFA:
-
non-esterified fatty acids
- NO2 :
-
nitrites
- NO3 :
-
nitrates
- NOx :
-
total nitric oxide metabolites
- PON:
-
paraoxonase
- ROMs:
-
total reactive oxygen metabolites.
References
ADAS (1986) Condition scoring of dairy cows. Publ. 612
Aitken SL, Karcher EL, Rezamand P et al (2009) Evaluation of antioxidant and proinflammatory gene expression in bovine mammary tissue during the periparturient period. J Dairy Sci 92:589–598. https://doi.org/10.3168/jds.2008-1551
Batistel F, Osorio JS, Tariq MR, Li C, Caputo J, Socha MT, Loor JJ (2017) Peripheral leukocyte and endometrium molecular biomarkers of inflammation and oxidative stress are altered in peripartal dairy cows supplemented with Zn, Mn, and Cu from amino acid complexes and Co from Co glucoheptonate. J Anim Sci Biotechnol 8:33. https://doi.org/10.1186/s40104-017-0163-7
Bertoni G, Trevisi E, Han X, Bionaz M (2008) Effects of inflammatory conditions on liver activity in puerperium period and consequences for performance in dairy cows. J Dairy Sci 91:3300–3310. https://doi.org/10.3168/jds.2008-0995
Bionaz M, Trevisi E, Calamari L, Librandi F, Ferrari A, Bertoni G (2007) Plasma paraoxonase, health, inflammatory conditions, and liver function in transition dairy cows. J Dairy Sci 90:1740–1750. https://doi.org/10.3168/jds.2006-445
Bionaz M, Periasamy K, Rodriguez-Zas SL et al (2012a) A novel dynamic impact approach (DIA) for functional analysis of time-course omics studies: validation using the bovine mammary transcriptome. PLoS One 7:e32455. https://doi.org/10.1371/journal.pone.0032455
Bionaz M, Periasamy K, Rodriguez-Zas SL, Everts RE, Lewin HA, Hurley WL, Loor JJ (2012b) Old and new stories: revelations from functional analysis of the bovine mammary transcriptome during the lactation cycle. PLoS One 7:e33268. https://doi.org/10.1371/journal.pone.0033268
Calamari L, Ferrari A, Minuti A, Trevisi E (2016) Assessment of the main plasma parameters included in a metabolic profile of dairy cow based on Fourier transform mid-infrared spectroscopy: preliminary results. BMC Vet Res 12:4. https://doi.org/10.1186/s12917-015-0621-4
Celi P, Gabai G (2015) Oxidant/antioxidant balance in animal nutrition and health: the role of protein oxidation. Front Vet Sci 2:48. https://doi.org/10.3389/fvets.2015.00048
Chomczynski P, Sacchi N (2006) The single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction: twenty-something years on. Nat Protoc 1:581–585. https://doi.org/10.1038/nprot.2006.83
Crookenden MA, Heiser A, Murray A et al (2016) Parturition in dairy cows temporarily alters the expression of genes in circulating neutrophils. J Dairy Sci 99:6470–6483. https://doi.org/10.3168/jds.2015-10877
Danussi C, Petrucco A, Wassermann B, Pivetta E, Modica TM, del Bel Belluz L, Colombatti A, Spessotto P (2011) EMILIN1–α4/α9 integrin interaction inhibits dermal fibroblast and keratinocyte proliferation. J Cell Biol 195:131–145. https://doi.org/10.1083/jcb.201008013
DeGaris PJ, Lean IJ (2008) Milk fever in dairy cows: a review of pathophysiology and control principles. Vet J 176:58–69. https://doi.org/10.1016/j.tvjl.2007.12.029
Drackley JK (1999) Biology of dairy cows during the transition period: the final frontier? J Dairy Sci 82:2259–2273. https://doi.org/10.3168/jds.S0022-0302(99)75474-3
Duffield TF, Lissemore KD, McBride BW, Leslie KE (2009) Impact of hyperketonemia in early lactation dairy cows on health and production. J Dairy Sci 92:571–580. https://doi.org/10.3168/jds.2008-1507
Goff JP, Horst RL (1997) Physiological changes at parturition and their relationship to metabolic disorders. J Dairy Sci 80:1260–1268. https://doi.org/10.3168/jds.S0022-0302(97)76055-7
Gordon JL, Leblanc SJ, Duffield TF (2013) Ketosis treatment in lactating dairy cattle. Vet Clin North Am Food Anim Pract 29:433–445. https://doi.org/10.1016/j.cvfa.2013.03.001
Granger DN, Kubes P (1994) The microcirculation and inflammation: modulation of leukocyte-endothelial cell adhesion. J Leukoc Biol 55:662–675
Hammon DS, Evjen IM, Dhiman TR, Goff JP, Walters JL (2006) Neutrophil function and energy status in Holstein cows with uterine health disorders. Vet Immunol Immunopathol 113:21–29. https://doi.org/10.1016/j.vetimm.2006.03.022
Hassan SS, Romero R, Haddad R, Hendler I, Khalek N, Tromp G, Diamond MP, Sorokin Y, Malone J Jr (2006) The transcriptome of the uterine cervix before and after spontaneous term parturition. Am J Obstet Gynecol 195:778–786. https://doi.org/10.1016/j.ajog.2006.06.021
Hodgkinson AJ, Carpenter EA, Smith CS et al (2007) Adhesion molecule expression in the bovine mammary gland. Vet Immunol Immunopathol 115:205–215. https://doi.org/10.1016/j.vetimm.2006.10.020
Jahan N, Minuti A, Trevisi E (2015) Assessment of immune response in periparturient dairy cows using ex vivo whole blood stimulation assay with lipopolysaccharides and carrageenan skin test. Vet Immunol Immunopathol 165:119–126. https://doi.org/10.1016/j.vetimm.2015.04.003
Kehrli ME, Goff JP (1989) Periparturient hypocalcemia in cows: effects on peripheral blood neutrophil and lymphocyte function. J Dairy Sci 72:1188–1196. https://doi.org/10.3168/jds.S0022-0302(89)79223-7
Kehrli ME, Nonnecke BJ, Roth JA (1989a) Alterations in bovine neutrophil function during the periparturient period. Am J Vet Res 50:207–214
Kehrli ME, Nonnecke BJ, Roth JA (1989b) Alterations in bovine lymphocyte function during the periparturient period. Am J Vet Res 50:215–220
Kimura K, Reinhardt TA, Goff JP (2006) Parturition and hypocalcemia blunts calcium signals in immune cells of dairy cattle. J Dairy Sci 89:2588–2595. https://doi.org/10.3168/jds.S0022-0302(06)72335-9
Kremer WDJ, Noordhuizen-Stassen EN, Grommers FJ, Schukken YH, Heeringa R, Brand A, Burvenich C (1993) Severity of experimental Escherichia coli mastitis in ketonemic and nonketonemic dairy cows. J Dairy Sci 76:3428–3436. https://doi.org/10.3168/jds.S0022-0302(93)77681-X
Lacetera N, Scalia D, Bernabucci U, Ronchi B, Pirazzi D, Nardone A (2005) Lymphocyte functions in overconditioned cows around parturition. J Dairy Sci 88:2010–2016. https://doi.org/10.3168/jds.S0022-0302(05)72877-0
Li B, Ruotti V, Stewart RM, Thomson JA, Dewey CN (2010) RNA-Seq gene expression estimation with read mapping uncertainty. Bioinformatics 26:493–500. https://doi.org/10.1093/bioinformatics/btp692
Littell RC, Henry PR, Ammerman CB (1998) Statistical analysis of repeated measures data using SAS procedures. J Anim Sci 76:1216–1231
Liu Y, Shaw SK, Ma S et al (2004) Regulation of leukocyte transmigration: cell surface interactions and signaling events. J Immunol 172:7–13
Loor JJ, Bertoni G, Hosseini A et al (2013) Functional welfare – using biochemical and molecular technologies to understand better the welfare state of peripartal dairy cattle. Anim Prod Sci 53:931–953. https://doi.org/10.1071/AN12344
Mallard BA, Dekkers JC, Ireland MJ, Leslie KE, Sharif S, Vankampen CL, Wagter L, Wilkie BN (1998) Alteration in immune responsiveness during the peripartum period and its ramification on dairy cow and calf health. J Dairy Sci 81:585–595
Morimoto T, Devora GA, Mibe M et al (1997) Parathyroid hormone-related protein and human myometrial cells: action and regulation. Mol Cell Endocrinol 129:91–99
Muller WA (2003) Leukocyte-endothelial-cell interactions in leukocyte transmigration and the inflammatory response. Trends Immunol 24:327–334
Nakamura MT, Yudell BE, Loor JJ (2014) Regulation of energy metabolism by long-chain fatty acids. Prog Lipid Res 53:124–144. https://doi.org/10.1016/j.plipres.2013.12.001
Pepino MY, Kuda O, Samovski D, Abumrad NA (2014) Structure-function of CD36 and importance of fatty acid signal transduction in fat metabolism. Annu Rev Nutr 34:281–303. https://doi.org/10.1146/annurev-nutr-071812-161220
Robinson MD, Oshlack A (2010) A scaling normalization method for differential expression analysis of RNA-seq data. Genome Biol 11:R25. https://doi.org/10.1186/gb-2010-11-3-r25
Sartorelli P, Paltrinieri S, Agnes F (1999) Non-specific immunity and ketone bodies. I: in vitro studies on chemotaxis and phagocytosis in ovine neutrophils. Zentralbl Veterinarmed A 46:613–619
Sartorelli P, Paltrinieri S, Comazzi S (2000) Non-specific immunity and ketone bodies. II: in vitro studies on adherence and superoxide anion production in ovine neutrophils. J Vet Med A Physiol Pathol Clin Med 47:1–8
Sheldon IM, Williams EJ, Miller ANA, Nash DM, Herath S (2008) Uterine diseases in cattle after parturition. Vet J 176:115–121. https://doi.org/10.1016/j.tvjl.2007.12.031
Silver DL, Tall AR (2001) The cellular biology of scavenger receptor class B type I. Curr Opin Lipidol 12:497–504
Sordillo LM, Aitken SL (2009) Impact of oxidative stress on the health and immune function of dairy cattle. Vet Immunol Immunopathol 128:104–109. https://doi.org/10.1016/j.vetimm.2008.10.305
Sordillo LM, Contreras GA, Aitken SL (2009) Metabolic factors affecting the inflammatory response of periparturient dairy cows. Anim Health Res Rev 10:53–63. https://doi.org/10.1017/S1466252309990016
Spessotto P, Cervi M, Mucignat MT, Mungiguerra G, Sartoretto I, Doliana R, Colombatti A (2003) β1 integrin-dependent cell adhesion to EMILIN-1 is mediated by the gC1q domain. J Biol Chem 278:6160–6167. https://doi.org/10.1074/jbc.M208322200
Spessotto P, Bulla R, Danussi C, Radillo O, Cervi M, Monami G, Bossi F, Tedesco F, Doliana R, Colombatti A (2006) EMILIN1 represents a major stromal element determining human trophoblast invasion of the uterine wall. J Cell Sci 119:4574–4584. https://doi.org/10.1242/jcs.03232
Suriyasathaporn W, Daemen AJ, Noordhuizen-Stassen EN, Dieleman SJ, Nielen M, Schukken YH (1999) Beta-hydroxybutyrate levels in peripheral blood and ketone bodies supplemented in culture media affect the in vitro chemotaxis of bovine leukocytes. Vet Immunol Immunopathol 68:177–186
Targowski SP, Klucinski W (1983) Reduction in mitogenic response of bovine lymphocytes by ketone bodies. Am J Vet Res 44:828–830
Targowski SP, Klucinski W, Littledike ET, Hoy DA (1985) Suppression of mitogenic response of bovine lymphocytes during experimental ketosis in calves. Am J Vet Res 46:1378–1380
Trapnell C, Pachter L, Salzberg SL (2009) TopHat: discovering splice junctions with RNA-Seq. Bioinformatics 25:1105–1111. https://doi.org/10.1093/bioinformatics/btp120
Trevisi E, Minuti A (2018) Assessment of the innate immune response in the periparturient cow. Res Vet Sci 116:47–54. https://doi.org/10.1016/j.rvsc.2017.12.001
Trevisi E, Amadori M, Bakudila AM, Bertoni G (2009) Metabolic changes in dairy cows induced by oral, low-dose interferon-alpha treatment. J Anim Sci 87:3020–3029. https://doi.org/10.2527/jas.2008-1178
Trevisi E, Zecconi A, Bertoni G, Piccinini R (2010) Blood and milk immune and inflammatory profiles in periparturient dairy cows showing a different liver activity index. J Dairy Res 77:310–317. https://doi.org/10.1017/S0022029910000178
Trevisi E, Amadori M, Cogrossi S, Razzuoli E, Bertoni G (2012) Metabolic stress and inflammatory response in high-yielding, periparturient dairy cows. Res Vet Sci 93:695–704. https://doi.org/10.1016/j.rvsc.2011.11.008
Trevisi E, Jahan N, Bertoni G et al (2015) Pro-inflammatory cytokine profile in dairy cows: consequences for new lactation. Ital J Anim Sci 14. https://doi.org/10.4081/ijas.2015.3862
Valacchi G, Sticozzi C, Lim Y, Pecorelli A (2011) Scavenger receptor class B type I: a multifunctional receptor. Ann N Y Acad Sci 1229:E1–E7. https://doi.org/10.1111/j.1749-6632.2011.06205.x
van Knegsel ATM, Hammon HM, Bernabucci U, Bertoni G, Bruckmaier RM, Goselink RMA, Gross JJ, Kuhla B, Metges CC, Parmentier HK, Trevisi E, Tröscher A, van Vuuren AM (2014). Metabolic adaptation during early lactation: key to cow health, longevity and a sustainable dairy production chain. CAB Reviews: Perspectives in Agriculture, Veterinary Science, Nutrition and Natural Resources, 9. https://doi.org/10.1079/PAVSNNR20149002
Wankhade PR, Manimaran A, Kumaresan A et al (2017) Metabolic and immunological changes in transition dairy cows: a review. Vet World 10:1367–1377. https://doi.org/10.14202/vetworld.2017.1367-1377
Wight TN, Raugi GJ, Mumby SM, Bornstein P (1985) Light microscopic immunolocation of thrombospondin in human tissues. J Histochem Cytochem 33:295–302. https://doi.org/10.1177/33.4.3884704
Wu WX, Zhang Q, Ma XH, Unno N, Nathanielsz PW (1999) Suppression subtractive hybridization identified a marked increase in thrombospondin-1 associated with parturition in pregnant sheep myometrium. Endocrinology 140:2364–2371. https://doi.org/10.1210/endo.140.5.6697
Zarrin M, Wellnitz O, van Dorland HA, Bruckmaier RM (2014) Induced hyperketonemia affects the mammary immune response during lipopolysaccharide challenge in dairy cows. J Dairy Sci 97:330–339. https://doi.org/10.3168/jds.2013-7222
Zhou Z, Bu DP, Vailati Riboni M, Khan MJ, Graugnard DE, Luo J, Cardoso FC, Loor JJ (2015) Prepartal dietary energy level affects peripartal bovine blood neutrophil metabolic, antioxidant, and inflammatory gene expression. J Dairy Sci 98:5492–5505. https://doi.org/10.3168/jds.2014-8811
Zimin AV, Delcher AL, Florea L, Kelley DR, Schatz MC, Puiu D, Hanrahan F, Pertea G, van Tassell C, Sonstegard TS, Marçais G, Roberts M, Subramanian P, Yorke JA, Salzberg SL (2009) A whole-genome assembly of the domestic cow, Bos taurus. Genome Biol 10:R42. https://doi.org/10.1186/gb-2009-10-4-r42
Acknowledgments
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
Funding
This study was funded by the “Romeo ed Enrica Invernizzi foundation,” Milan, Italy.
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Ethics approval and consent to participate
All procedures performed in studies involving animals were in accordance with the ethical standards of the institution at which the studies were conducted (DL No. 116, 27/01/1992)
Conflict of interest
The authors declare that they have no conflict of interest.
Additional information
Publisher’s note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
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
Received:
Revised:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10142-019-00720-0