Methionine supply during the peripartum period and early lactation alter immunometabolic gene expression in cytological smear and endometrial tissue of holstein cows.

The objective of the present study was to evaluate the effect of feeding rumen-protected methionine (RPM) during the peripartal period and early lactation on mRNA gene expression profiles of uterine cytological smear and endometrial samples of Holstein cows (n = 20). Treatments consisted of a supplementation with RPM [MET; n = 11; RPM at a rate of 0.08 % of DM: Lys:Met = 2.8:1, (Smartamine® M Adisseo, Alpharetta, GA, USA)] and no supplementation (CON; n = 9; Lys:Met = 3.5:1). Uterine cytology smears and endometrial samples were collected at 15, 30, and 73 days in milk (DIM) and analyzed for expression of genes related with metabolism, inflammation, and methionine metabolism. Regarding the cytological smear samples, RPM supplementation tended to increase mRNA expression of methionine adenosyltransferase 1 alpha (MAT1A) and increased the mRNA expression of fibroblast growth factor 7 (FGF7), with an effect of time for the latter. On the other hand, RPM decreased mRNA expression for glucose transporter 4 (GLUT4), interleukin 1 beta (IL-1ß), interleukin 6 (IL-6), interleukin 8 (IL-8), prostaglandin E synthase 3 (PTGES3), translocator protein 18 kDa (TSPO), mucin 1 (MUC1) and superoxide dismutase (SOD1) in cytological smear samples. There was an effect of time for all variables except MAT1A, with decreasing expression over time. There was a TRT × TIME interaction for GLUT4 mRNA expression, with higher GLUT4 mRNA expression for cows fed CON than for cows fed RPM at time 15 and a tendency to higher expression for cows fed CON on time 30 when compared with cows fed RPM. For uterine tissue samples, feeding RPM increased the mRNA expression of lecithin-cholesterol acyltransferase (LCAT), S-adenosyl-l-homocysteine hydrolase (SAAH), FGF7, GLUT4, and apolipoproteins 3 (APOL3), with an effect of time for APOL3 where its expression increased over time. There was a tendency for cows fed RPM to have decreased IL1ß mRNA expression. In conclusion, feeding RPM during transition period and early lactation is beneficial for uterine immune response and metabolism in early lactation as indicated by the favorable expressions of genes affecting the uterine immunometabolism during such a challenging period.

Improved uterine immune mediators in Holstein cows supplemented with rumen-protected methionine and discovery of neutrophil extracellular traps (NET).

During the transition from prepartum to early lactation, dairy cows often experience negative energy balance (NEB) that may result in reproductive stress and decreased fertility. The objective of this study was to observe the effects 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. 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). Uterine 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 percentage. Cows in CON had lower methionine plasma concentrations (18.05 ±â€¯2.0 µM) than cows in MET (30.39 ±â€¯1.6 µM). Cows in CON had greater cystine plasma concentrations (3.62 ±â€¯0.3 µM) than cows in MET (2.8 ±â€¯0.3 µM). No treatment differences were observed for SOD or GPX in the endometrium. Cows in CON tended to have a high score for positively immunolabeled GPX cells at 15 DIM than cows in MET. No treatment differences were observed for the percentage of PMN in uterine cytology, number of neutrophils, or extent of NET formation in the endometrium. A treatment by time interaction was observed for PMN percentage and the number of neutrophils: cows in MET tended to have greater PMN percentages 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.