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Title:Uterine immune mediators and lipid profiles of the preimplantation embryo and endometrial tissue in Holstein cows supplemented with rumen-protected methionine and observed neutrophil extracellular trap (NET) formation in the bovine endometrial tissue
Author(s):Stella, Stephanie Lynn
Advisor(s):Cardoso, Phil
Contributor(s):Drackley, James; Wheeler, Matthew; Steelman, Andrew
Department / Program:Animal Sciences
Discipline:Animal Sciences
Degree Granting Institution:University of Illinois at Urbana-Champaign
Degree:M.S.
Genre:Thesis
Subject(s):Methionine
neutrophil extracellular trap
uterine immune function
Abstract:Future reproductive success of the dairy industry continues to decline as the push for high-producing dairy cows increases. The transition from gestation into early lactation can be stressful for the dairy cow and result in low fertility in the future. Implementation of feed strategies can improve the transition from parturition to lactation for the cow, both metabolically and physically, and possibly reduce the risk of postpartum uterine diseases and early embryonic loss. Two experiments were conducted in order to observe the effects of nutritional supplementation on reproductive status and future embryonic development from the transition period to early lactation in Holstein cows. To investigate the effect of rumen-protected methionine (RPM) on plasma amino acid concentrations, uterine cytology, immunohistochemistry (IHC) of glutathione peroxidase 1 (GPX) and superoxide dismutase 1 (SOD), and to confirm neutrophil extracellular trap (NET) formation in the endometrial tissue, multiparous Holstein cows (n = 20) were randomly assigned to two treatments starting at 21 d before calving until 73 days in milk (DIM). Treatments were: CON (n = 9, no supplementation, TMR with a Lys:Met = 3.5:1) and MET (n = 11, TMR + Smartamine® M with a Lys:Met = 2.8:1). Endometrial biopsies, uterine cytology, and blood samples from the coccygeal artery or vein were collected at 15, 30, and 73 DIM. Blood plasma samples were analyzed for amino acids and metabolites. Uterine biopsies were analyzed for NET formation, neutrophil numbers, as well as GPX and SOD by IHC. Additionally, uterine cytology was analyzed for polymorphonuclear neutrophil (PMN) to epithelial cell proportion. Cows in CON had lower (P < 0.01) methionine plasma concentrations than cows in MET. Cows in CON had greater (P < 0.01) cystine plasma concentrations than cows in MET. No treatment differences (P > 0.10) were observed for SOD or GPX in the endometrium. Cows in CON tended to have a higher score (P = 0.08) for positively immunolabeled GPX cells at 15 DIM than cows in MET. No treatment differences (P > 0.10) were observed for the proportion of PMN in uterine cytology, number of neutrophils, or extent of NET formation in the endometrium. A treatment by time interaction (P = 0.06) was observed for PMN proportion and the number of neutrophils: cows in MET tended to have greater PMN proportion than cows in CON at 15 DIM which decreased for subsequent days and cows in MET had greater neutrophil numbers in the endometrium at 30 DIM than cows in CON. In conclusion, dietary supplementation of RPM altered plasma amino acid concentrations and increased neutrophil infiltration in the postpartum period, suggesting improved uterine immunity. Furthermore, we examined the effects of rumen-protected methionine on the lipid profiles of the preimplantation embryo and endometrial tissue. Multiparous Holstein cows (n = 20) were randomly assigned to two treatments starting at 21 d before calving until 73 days in milk (DIM). Treatments were: CON (n = 9, no supplementation, TMR with a Lys:Met = 3.5:1) and MET (n = 11, TMR + Smartamine® M with a Lys:Met = 2.8:1). Endometrial biopsies were obtained at 15, 30, and 73 DIM and preimplantation embryos were harvested at 73 DIM. Endometrial lipid profiles were analyzed using multiple reaction monitoring-profiling and lipid profiles of embryos were analyzed using matrix assisted laser desorption/ionization mass spectrometry. Cows in MET had increased (P ≤ 0.05) polyunsaturated lipid concentrations in embryos than cows in CON. Cows in MET had decreased (P < 0.10) monounsaturated lipid concentrations in embryos than cows in CON. Cows in CON had increased (P < 0.14) concentrations of saturated lipids in uterine tissue at 15 DIM than cows in MET. Cows in CON had increased (P < 0.13) concentrations of un/monounsaturated lipids in uterine tissue at 15 DIM than cows in MET. Cows in MET had increased (P < 0.10) saturated lipid concentrations in uterine tissue at 30 DIM than cows in CON. Cows in MET had increased (P ≤ 0.05) concentrations of unsaturated lipids in uterine tissue at 30 DIM than cows in CON. Cows in MET had increased (P ≤ 0.05) monounsaturated lipid concentrations in uterine tissue at 30 DIM than cows in CON. Concentrations of saturated lipids in uterine tissue at 73 DIM were increased (P = 0.06) for cows in MET than cows in CON. Cows in MET had increased (P = 0.10) concentrations of unsaturated lipids in uterine tissue at 73 DIM than cows in CON. Finally, cows in MET had increased (P ≤ 0.05) monounsaturated lipid concentrations in uterine tissue at 73 DIM than cows in CON. In conclusion, lipid profiles of the preimplantation embryo and endometrial tissue of cows fed RPM were altered, which may be indicative of improved embryonic vitality. The supplementation of RPM may improve reproductive success of dairy cows during and after the transition period, as well as reduce embryonic death in the future.
Issue Date:2017-07-19
Type:Thesis
URI:http://hdl.handle.net/2142/98398
Rights Information:© 2017 Stephanie Lynn Stella
Date Available in IDEALS:2017-09-29
Date Deposited:2017-08


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