Department of Animal Sciences, University of Illinois, Urbana, IL, USA; The Colombian Corporation for Agricultural Research (CORPOICA), Bogotá, Colombia; Department of Clinics, Faculty of Veterinary Medicine, Universidade Federal de Pelotas, Pelotas, RS, Brazil.
Department of Animal Sciences, University of Illinois, Urbana, IL, USA.
Theriogenology. 2017 Jul 1;96:1-9. doi: 10.1016/j.theriogenology.2017.03.022. Epub 2017 Mar 27.
Multiparous Holstein cows were assigned in a randomized complete block design into four treatments from 21 d before calving to 30 d in milk (DIM). Treatments were: MET [n = 19, fed the basal diet + rumen-protected methionine at a rate of 0.08% (w/w) of the dry matter, Smartamine M], CHO (n = 17, fed the basal diet + choline 60 g/d, Reashure), MIX (n = 21, fed the basal diet + Smartamine M at a rate of 0.08% (w/w) of the dry matter and 60 g/d Reashure), and CON (n = 20, no supplementation, fed the close-up and fresh cow diets). Follicular development was monitored via ultrasound every 2 d starting at 7 DIM until ovulation (n = 37) or aspiration (n = 40) of the first postpartum dominant follicle (DF). Follicular fluid from 40 cows was aspirated and cells were retrieved immediately by centrifugation. Gene expression of TLR4, TNF, IL1-β, IL8, IL6, LHCGR, STAR, 3β-HSD, P450scc, CYP19A1, IRS1, IGF, MAT1A, and SAHH, was measured in the follicular cells of the first DF. Cows in CON had higher TNF, TLR4, and IL1-β mRNA expression (11.70 ± 4.6, 21.29 ± 10.4, 6.28 ± 1.4, respectively) than CHO (2.77 ± 0.9, 2.16 ± 0.9, 2.29 ± 0.7, respectively), and MIX (2.23 ± 0.7, 1.46 ± 0.6, 2.92 ± 0.8, respectively). Cows in CON had higher IL1-β expression (6.27 ± 1.4) than cows in MET (3.28 ± 0.6). Expression of IL8 mRNA was lower for cows in CHO (0.98 ± 0.3) than cows in CON (4.90 ± 0.7), MET (6.10 ± 1.7), or MIX (5.05 ± 1.8). Treatments did not affect mRNA expression of LHCGR, STAR, P450scc, CYP19A, SAHH, MAT1A, or IL6 however, 3β-HSD expression was higher for cows in MET (1.46 ± 0.3) and MIX (1.25 ± 0.3) than CON (0.17 ± 0.04) and CHO (0.26 ± 0.1). Supplementation of methionine, choline, and both methionine and choline during the transition period did not affect days to first ovulation or number of cows that ovulated the first follicular wave. Plasma and follicular fluid estradiol and progesterone concentrations were not different among treatments. Methionine concentrations in the follicular fluid of the first postpartum DF was higher for cows in MET (18.2 ± 0.1 μM) than cows in CON (11.1 ± 0.9 μM). In conclusion, supplementing choline and methionine during the transition period changed mRNA expression in follicular cells and dietary methionine supplementation increased plasma and follicular fluid concentrations of methionine of the first postpartum DF in Holstein cows.
多胎荷斯坦奶牛在分娩前 21 天至泌乳后 30 天（DIM）期间，根据随机完全区组设计分为四组：MET[ n = 19，饲喂基础日粮+保护型蛋氨酸，添加量为干物质的 0.08%（w/w），Smartamine M]、CHO[ n = 17，饲喂基础日粮+60g/d 胆碱，Reashure]、MIX[ n = 21，饲喂基础日粮+0.08%（w/w）的 Smartamine M 和 60g/d Reashure]和 CON[ n = 20，不补充，饲喂接近牛和新产牛日粮]。从 7DIM 开始，每隔 2d 通过超声监测卵泡发育，直到排卵（n = 37）或抽吸（n = 40）第一头产后优势卵泡（DF）。从 40 头牛中抽吸卵泡液，立即通过离心回收细胞。测量第一头 DF 中卵泡细胞 TLR4、TNF、IL1-β、IL8、IL6、LHCGR、STAR、3β-HSD、P450scc、CYP19A1、IRS1、IGF、MAT1A 和 SAHH 的基因表达。与 CHO（2.77 ± 0.9、2.16 ± 0.9、2.29 ± 0.7）和 MIX（2.23 ± 0.7、1.46 ± 0.6、2.92 ± 0.8）相比，CON 中的奶牛 TNF、TLR4 和 IL1-β mRNA 表达更高（分别为 11.70 ± 4.6、21.29 ± 10.4 和 6.28 ± 1.4）。与 MET（3.28 ± 0.6）相比，CON 中的奶牛 IL1-β 表达更高（6.27 ± 1.4）。与 CON（4.90 ± 0.7）、MET（6.10 ± 1.7）或 MIX（5.05 ± 1.8）相比，CHO 中的奶牛 IL8 mRNA 表达较低（0.98 ± 0.3）。然而，LHCGR、STAR、P450scc、CYP19A、SAHH、MAT1A 或 IL6 的 mRNA 表达不受处理影响，但 MET（1.46 ± 0.3）和 MIX（1.25 ± 0.3）中的奶牛 3β-HSD 表达高于 CON（0.17 ± 0.04）和 CHO（0.26 ± 0.1）。在过渡期补充蛋氨酸、胆碱和蛋氨酸和胆碱都没有影响第一排卵或排卵第一卵泡波的牛的数量。处理之间的血浆和卵泡液雌二醇和孕酮浓度没有差异。MET 中的奶牛第一产后 DF 中的蛋氨酸浓度较高（18.2 ± 0.1 μM）比 CON 中的奶牛（11.1 ± 0.9 μM）。总之，在过渡期补充胆碱和蛋氨酸改变了卵泡细胞的 mRNA 表达，而日粮蛋氨酸补充增加了荷斯坦奶牛产后第一 DF 的血浆和卵泡液蛋氨酸浓度。
