title: Effects of rumen-protected methionine and choline supplementation on the preimplantation embryo in Holstein cows.
abstract: Our objective was to determine the effects of supplementing methionine and choline during the prepartum and postpartum periods on preimplantation embryos of Holstein cows. Multiparous cows were assigned in a randomized complete-block design into four treatments from 21 days before calving to 30 days in milk (DIM). Treatments (TRT) were MET (n = 9, fed the basal diet + rumen-protected methionine at a rate of 0.08% [w:w] of the dry matter [DM], Smartamine M), CHO (n = 8, fed the basal diet + choline 60 g/d, Reashure), MIX (n = 11, fed the basal diet + Smartamine M and 60 g/d Reashure), and CON (n = 8, no supplementation, fed the close-up and fresh cow diets). Cows were randomly reassigned to two new groups (GRP) to receive the following diets from 31 to 72 DIM; control (CNT, n = 16, fed a basal diet) and SMT (n = 20, fed the basal diet + 0.08% [w:w] of the dry matter intake as methionine). An progesterone intravaginal insert (CIDR) device was inserted in all cows after follicular aspiration (60 DIM) and superovulation began at Day 61.5 using FSH in eight decreasing doses at 12-hour intervals over a 4-day period. On Days 63 and 64, all cows received two injections of PGF2α, and CIDR was removed on Day 65. Twenty-four hours after CIDR removal, ovulation was induced with GnRH. Cows received artificial insemination at 12 hours and 24 hours after GnRH. Embryos were flushed 6.5 days after artificial insemination. Global methylation of the embryos was assessed by immunofluorescent labeling of 5-methylcytosine, whereas lipid content was assessed by staining with Nile red. Nuclear staining was used to count the total number of cells per embryo. There was no difference between TRT, GRP, or their interaction (P > 0.05) for embryo recovery, embryos recovered, embryo quality, embryo stage, or cells per embryo. Methylation of the DNA had a TRT by GRP interaction (P = 0.01). Embryos from cows in CON-CNT had greater (P = 0.04) methylation (0.87 ± 0.09 arbitrary units [AU]) than embryos from cows in MET-CNT (0.44 ± 0.07 AU). The cytoplasmic lipid content was not affected (P > 0.05) by TRT or their interaction, but lipid content was greater (P = 0.04) for SMT (7.02 ± 1.03 AU) than that in CNT (3.61 ± 1.20 AU). In conclusion, cows in MET-CNT had embryos with lower methylation, and SMT cows had a higher lipid content than CNT. Methionine supplementation seems to impact the preimplantation embryo in a way that enhances its capacity for survival because there is strong evidence that endogenous lipid reserves serve as an energy substrate.
authors.affiliation: Department of Animal Sciences, University of Illinois, Urbana, Illinois, USA; Faculty of Veterinary Medicine, Department of Clinics, Universidade Federal de Pelotas, Pelotas, Rio Grande do Sul, Brazil.
authors.forename: D A V
authors.initials: DAV
authors.lastname: Acosta
authors.affiliation: Department of Biology, Northeastern University, Boston, Massachussets, USA; Department of Animal Science, University of Florida, Gainesville, Florida, USA.
authors.forename: A C
authors.initials: AC
authors.lastname: Denicol
authors.affiliation: Department of Animal Science, University of Florida, Gainesville, Florida, USA.
authors.forename: P
authors.initials: P
authors.lastname: Tribulo
authors.affiliation: Department of Animal Sciences, University of Illinois, Urbana, Illinois, USA.
authors.forename: M I
authors.initials: MI
authors.lastname: Rivelli
authors.affiliation: Department of Animal Sciences, University of Illinois, Urbana, Illinois, USA.
authors.forename: C
authors.initials: C
authors.lastname: Skenandore
authors.affiliation: Department of Animal Sciences, University of Illinois, Urbana, Illinois, USA.
authors.forename: Z
authors.initials: Z
authors.lastname: Zhou
authors.affiliation: Adisseo NACA, Alpharetta, Georgia, USA.
authors.forename: D
authors.initials: D
authors.lastname: Luchini
authors.affiliation: Faculty of Veterinary Medicine, Department of Clinics, Universidade Federal de Pelotas, Pelotas, Rio Grande do Sul, Brazil.
authors.forename: M N
authors.initials: MN
authors.lastname: Corrêa
authors.affiliation: Department of Animal Science, University of Florida, Gainesville, Florida, USA.
authors.forename: P J
authors.initials: PJ
authors.lastname: Hansen
authors.affiliation: Department of Animal Sciences, University of Illinois, Urbana, Illinois, USA. Electronic address: cardoso2@illinois.edu.
authors.forename: F C
authors.initials: FC
authors.lastname: Cardoso
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000187:drug effects*; D002417:Cattle / Q000196:embryology*; D002794:Choline / Q000627:therapeutic use*; D019175:DNA Methylation / Q000187:drug effects; D019587:Dietary Supplements; D047108:Embryonic Development / Q000187:drug effects; D050356:Lipid Metabolism; D008715:Methionine / Q000627:therapeutic use*
chemical_list: D008715:Methionine; D002794:Choline
keywords: Choline; Embryo; Lipid; Methionine; Methylation