Rumen-protected Methionine Supplementation Research Articles

Rumen-protected methionine supplementation during the peripartal period alters the expression of galectin genes associated with inflammation in peripheral neutrophils and secretion in plasma of Holstein cows.

The work described in this research communication aimed to investigate whether rumen-protected methionine (Met) supplementation during the periparturient period would affect the expression of galectins in blood-derived neutrophils, and secretion of galectins, IL (interleukin)-1β, IL-6, myeloperoxidase (MPO), and glucose in plasma. Because supplementation of rumen-protected Met would alleviate inflammation and oxidative stress during the peripartal period, we hypothesized that enhancing Met supply would benefit the innate immune response at least in part by altering the expression of galectin genes associated with neutrophil activity and inflammation. Galectins (Gal) have an immuno-modulating effect acting like cell-surface receptors whose activation often results in signaling cascades stimulating cells such as neutrophils. This study revealed an association between Met supplementation and galectin expression and secretion. This implies that galectin expression and secretion can be modulated by Met supplementation. Further studies are needed to evaluate the regulation of galectin gene expression for therapeutic and dietary intervention in the peripartal cow.

Beneficial effects of rumen-protected methionine on nitrogen-use efficiency, histological parameters, productivity and reproductive performance of ruminants

Providing essential amounts of balanced nutrients is one of the most vital aspects of livestock production. Among nutrients, protein has an essential role in many physiological functions of animals. Amino acids in needs for both high and medium yielding ruminant animals are not fully covered by microbial degraded feed sources in the rumen of animals, and they must be met by protecting the proteins from being broken down in the rumen; hence, the dietary supplementation of rumen-protected proteins (RPP), including mainly rumen-protected methionine (RPM), became imperative. Many researchers are interested in studying the role of (RPM) in ruminant animals concerning its effect on milk yield, growth performance, digestibility, dry matter intake and nitrogen utilization efficiency. Unfortunately, results obtained from several investigations regarding RPM indicated great fluctuation between its useful and useless effects in ruminant nutrition particularly during early and late lactation period; therefore, this review article may be helpful for ruminant farm owners when they decide to supplement RPM in animal’s diet. Conclusively, supplementation of RPM often has a balanced positive influence, without any reported negative impact on milk yield, growth performance and blood parameters especially in early lactating ruminant animals and when used with the low crude protein diet.

Supplementing rumen-protected methionine to lactating multiparous dairy cows did not improve reproductive performance.

There is evidence that supplementing methionine has positive effects on uterine environment, oocyte quality and embryo development in cattle. Thus, the objective of this study was to evaluate reproductive traits of cows supplemented with rumen‐protected methionine (RPM) during early to mid‐lactation in comparison with an untreated control group (CON). An additional focus was on the effect of puerperal diseases on reproductive performance parameters in RPM‐supplemented group MET and in CON. A total of 1,709 multiparous Holstein‐Friesian cows were enrolled in this field trial conducted on a commercial dairy farm in Slovakia. Cows were allocated at approximately 12 days post‐partum (dpp) to either CON or MET, the latter supplemented with 25.0 g–27.2 g RPM per cow per day incorporated into the total mixed ration (TMR) until leaving the study pen at approximately 140 dpp. The amount of RPM was calculated based on individual feed ingredients analysis and adjusted during the study period when TMR changed. Cows were monitored during the post‐partum period by vaginal examination (day 5 pp), measuring of beta‐hydroxybutyrate in blood (3, 5, and 8 dpp) and by vaginal examination, uterine cytology and measuring of back fat thickness by ultrasound (all at 31 ± 3 dpp).Compared with CON, cows supplemented with RPM did not show better reproduction performance parameters (first service submission rate, days to first service, conception risk, days open 140). Results from binary logistic regression model for the risk of conception showed that metritis had a significant effect, but the supplementation of methionine had not. Results of Cox regression analysis for the odds of conception within 140 dpp revealed only metritis and clinical endometritis as significant factors. In conclusion, supplementation of RPM had no beneficial effect on reproductive performance in this study farm compared with an untreated control group.

Effect of rumen-protected methionine supplementation to diets of Holstein dairy cows on their lactation performance

The objective of the present study was to evaluate the effect of rumen-protected methionine supplementation to diets of high producing Holstein dairy cows on milk yield and milk chemical composition. Ninety-nine pregnant Holstein dairy cows were randomly assigned to three nutritional groups to receive a total mixed ration and supplemented with rumen protected methionine (RPM) as; Smartamin (T1) or Met55 (T2) or non-supplemented (the control), at 260 days late gestation. Although, the two treated groups, (T1 and T2) were daily supplemented with RPM for 30 days pre- and another 30 days post-partum; different experimental groups including the control one were daily provided with 60 g/h choline chloride (Reashour) and 300 cm3 propylene glycol/ h, as an additional supply, during the same treating periods. Different experimental groups were fed a basal ration (16.2 CP % and 70 TDN %) during pregnancy and (16.4 CP % and 74 TDN %) during lactation. Results obtained showed that, supplementing different experimental groups with either Smartamin or Met55 did not improve (p<0.05) cows actual milk yield, 3.5%, fat corrected milk (FCM), energy corrected milk (ECM), milk fat yield nor protein yield kg.h.d-1. Supplementing Holstein dairy cows with (RPM) did not lead to any significant improvement (p<0.05), neither in animals feed conversion ratio, nor milk chemical composition traits. Animals body condition score (BCS) differed (p<0.05) among different experimental groups due to their yielding productivity in the former breeding season. However, lower milking group tended to have an improved BCS (3.79), but without significant difference with highly producing ones. In conclusion, Smartamin as (RPM) supplement was favored than that of Met55, if there were a necessity to be provided to lactating cows under the local environment.

Changes in growth performance, carcass characteristics, and meat properties of late fattening Hanwoo steers according to supplementation of rumen protected methionine and lysine

Changes in growth performance, carcass characteristics, and meat properties of late fattening Hanwoo steers according to supplementation of rumen protected methionine and lysine

Maternal supply of methionine during late pregnancy is associated with changes in immune function and abundance of microRNA and mRNA in Holstein calf polymorphonuclear leukocytes

Pregnancy and early life are critical periods during which environmental factors such as nutrition can affect development. Rumen-protected methionine (Met; RPM) supplementation during the prepartum period improves not only performance but immune responses in dairy cows. We investigated the effects of enhanced maternal supply of Met via feeding RPM on whole-blood in vitro lipopolysaccharide (LPS; 0, 0.01, or 5 μg/mL of blood) challenge and targeted microRNA and mRNA abundance in calf blood polymorphonuclear leukocytes (PMNL). Calves (n = 12/maternal diet) born to cows fed RPM at 0.08% of diet dry matter (DM)/d (MET) for the last 21 ± 2 d before calving or fed a control diet with no added Met (CON) were used. The PMNL were isolated at birth (before colostrum feeding) and d 1 (24 h after colostrum intake), 14, 28, and 50 of age. Maternal blood was collected at -10 ± 1.3 d relative to calving. Cows in the MET group had greater DM intake and lower prepartal haptoglobin concentration. In CON cows, haptoglobin was positively correlated with proinflammatory and host-defense mRNA abundance in CON calves. Except for NOS2 and NFE2L2, abundance of CASP8, MPO, ZBP1, and TNF was lower at birth in MET calves. Interleukin 1β concentration in response to LPS challenge in CON and MET calves was greatest at birth, underscoring the role of this cytokine for lymphocyte activation. Compared with 1 d of age, the interleukin-1β response to incremental doses of LPS was greater at 14 through 28 d, suggesting that the neonatal calf can mount a robust response to inflammatory stimuli. Greater abundance in CON calves of NOS2, CADM1, and TLR2 coupled with lower SELL from 1 through 50 d of age suggested a chronic activation of the PMNL. There was a marked upregulation over time of MIR125b, MIR146a, MIR155, and MIR9 in both CON and MET calves, suggesting that these microRNA could affect gene transcription associated with differentiation and inflammatory function in PMNL. Regardless of maternal diet, the gradual downregulation of MIR223 (the most abundant microRNA in PMNL) is in line with the progressive increase over time in the proinflammatory signature of the PMNL. Data revealed the potential for maternal supply of Met during late pregnancy through either greater DM intake or Met to elicit some changes in PMNL function during early postnatal life, partly through changes in mRNA expression encompassing cell adhesion and chemotaxis, oxidative stress, Toll-like receptor signaling, and Met metabolism.

Post-ruminal effects of rumen-protected methionine supplementation with low protein diet using long-term simulation and in vitro digestibility technique

Microbial degradation in the rumen and dietary availability of methionine amino acid have been reported as limiting in dairy ruminants. The aim of the present study was to examine the post-ruminal effects of feeding ruminants different concentrations of rumen-protected methionine (RPM) in low crude protein diets using the long-term rumen simulation method (Rusitec) followed by in vitro abomasum and ileum digestibility technique. The experiment contained four treatment groups: (1) high protein, without RPM supplementation (HP); (2) low protein, without RPM supplementation (LP); (3) low protein supplementation with low RPM (LPLM); and (4) low protein supplementation with high RPM (LPHM) mixed per 20 ± 0.04 g basal diet in every fermenter. The results showed that the LPLM and LPHM groups had significantly higher disappearance of crude protein and neutral detergent fiber in the abomasum and ileum than the HP treatment (P < 0.05) and were the same as the LP group (P > 0.05). The proportions of short-chain fatty acids and total volatile fatty acids in the abomasum and ileum were the same between the LPHM and HP groups (P > 0.05); however, the LPLM group was found to be significantly (P < 0.05) lower than the HP group and similar to the LP group (P > 0.05). Rusitec pH before or after changing feed bags and daily ammonia nitrogen production in the abomasum and ileum were non significantly (P > 0.05) different among all groups. In conclusion, RPM supplementation with low crude protein diets promoted post-ruminal digestibility and production of volatile fatty acids.

Lactation response to soybean meal and rumen-protected methionine supplementation of corn silage-based diets

Corn silage, an important forage fed to dairy cows in the United States, is energy rich but protein poor. The objectives of this experiment were to investigate the effects on production of milk and milk components of feeding corn silage-based diets with 4 levels of dietary crude protein (CP) plus rumen-protected methionine (RPM). Thirty-six cows were blocked by days in milk into 9 squares and randomly assigned to 9 balanced 4 × 4 Latin squares with four 4-wk periods. All diets were formulated to contain, as a percent of dry matter (DM), 50% corn silage, 10% alfalfa silage, 4% soyhulls, 2.4% mineral-vitamin supplement, and 30% neutral detergent fiber. Supplemental RPM (Mepron, Evonik Corp., Kennesaw, GA) was added to all diets to maintain a Lys:Met ratio of 3.1 in digested AA. Ground high-moisture corn was reduced and soybean meal (SBM) plus RPM increased to give diets containing, on average, 11% CP (28% corn, 31% starch, 6% SBM, 4 g of RPM/d), 13% CP (23% corn, 29% starch, 10% SBM, 8 g of RPM/d), 15% CP (19% corn, 26% starch, 15% SBM, 10 g of RPM/d), and 17% CP (14% corn, 24% starch, 19% SBM, 12 g of RPM/d). Data from the last 14 d of each period were analyzed using the mixed procedures in SAS (SAS Institute Inc., Cary, NC). With the exception of milk fat and milk lactose content, we found no significant effects of diet on all production traits. We did note linear responses to dietary CP concentration for intake, production of milk and milk components, and MUN. Cows fed the 11% CP diet had reduced DM intake, lost weight, and yielded less milk and milk components. Mean separation indicated that only true protein yield was lower on 13% CP than on 17% dietary CP, but not different between 15 and 17% CP. This indicated no improvement in production of milk and milk components above 15% CP. Quadratic trends for yield of milk, energy-corrected milk, and true protein suggested that a dietary CP concentration greater than 15% may be necessary to maximize production or, alternately, that a plateau was reached and no further CP was required. Although diet influenced apparent digestibility of DM, organic matter, and neutral detergent fiber, digestibility did not increase linearly with dietary CP. However, we observed linear and quadratic effects of dietary CP on acid detergent fiber digestibility. As expected, we found a linear effect of dietary CP on apparent N digestibility and on fecal and urinary N excretion, but no effect of diet on estimated true N digestibility. Ruminal concentrations of ammonia, total AA, peptides, and branched-chain volatile fatty acids also increased linearly with dietary CP. Quadratic responses indicated that 14.0 to 14.8% CP was necessary to optimize digestion and energy utilization. Overall results indicated that, when RPM was added to increase Lys:Met to 3.1, 15% CP was adequate for lactating dairy cows fed corn silage diets supplemented with SBM and secreting about 40 kg of milk/d; N excretion was lower than at 17% CP but with no reduction in yield of milk and milk components.

EFFECT OF PROTECTED METHIONINE SUPPLEMENTATION ON PRODUCTIVE PERFORMANCE OF GROWING BUFFALO CALVES

Fifteen male buffalo calves with an average of initial live body weight ranged between 250 and 267 kg were used to study the effect of rumen-protected methionine (RPM) supplementation on growth performance, nutrients digestibility and some ruminal parameters of growing male buffalo calves. Calves were randomly assigned into three nutritional groups (each of five animals) to receive one of the following experimental rations: the first group served as a control (T1) which received the basal ration (without methionine supplementation); (T2) received the basal ration (T1) and was supplemented with 15 g RPM /h /d; and (T3) received the basal ration (T1) plus 25 g RPM /h /d. Results obtained indicated that different experimental rations had in general almost similar chemical composition. Adding RPM to basal ration of buffalo calves, led to significant improvement (P 0.05) in different ruminal parameters, due to RPM supplement to the basal ration, while time of sampling indicated significant differences (P<0.05) for pH, NH3-N and TVFA's values at different times. Data obtained pointed out also to positive insignificant impact of RPM supplementation on accelerating calves daily gain and FCR. Efficiencyof feed utilization (FCR) was highest in T3 followed by T2in compare with T1, however, differences were statistically non-significant (P<0.05).

Effects of rumen-protected methionine and choline supplementation on vaginal discharge and uterine cytology of Holstein cows

Methionine is one of the most limiting amino acids in dairy diets and low feed intake around the time of calving could lead to decreased synthesis of phosphatidylcholine. An alternative pathway for phosphatidylcholine is to have choline as a precursor. The objective of this study was to determine the effects of feeding rumen-protected methionine and choline pre – and postpartum on reproduction of Holstein cows. Seventy-two Holstein cows were randomly assigned to four treatments from 21days before calving to 30days in milk (DIM): supplementation with rumen-protected methionine (MET; n=20, received 0.08% of the dry matter (DM) of the diet/d as methionine, Smartamine M® to a Lys:Met=2.9:1), rumen-protected choline (CHO; n=17, received 60g/d choline, Reassure), both rumen protected methionine and choline (MIX; n=19, received 0.08% of the DM of the diet/d as methionine to a Lys:Met=2.9:1 and 60g/d choline), or no supplementation to serve as control (CON; n=16, fed total mixed ration with a Lys:Met=3.5:1). Cows were evaluated at 4, 7, 10, 13, 15, 17, and 30d after calving for the presence of secretion using the Metricheck® device. On 15, 30, and 72d after calving, the uterine endometrium of all cows was sampled using a cytological brush and streaked onto slides for analysis of the presence of polymorphonuclear neutrophils (PMN). We hypothesized that cows supplemented with methionine would have lower metricheck smell scores and lower rates of PMN than non-supplemented cows. On d 30, a treatment difference was detected using the metricheck score and smell (P<0.04), with treatment MIX (score=0.38) having a lower score than CHO (score=2.11). Supplementing cows with rumen-protected methionine may have a beneficial effect on cows’ uterine health.

Effects of rumen-protected methionine and choline supplementation on steroidogenic potential of the first postpartum dominant follicle and expression of immune mediators in Holstein cows

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.

Expression of fatty acid sensing G-protein coupled receptors in peripartal Holstein cows

BackgroundG-protein coupled receptors (GPCR), also referred as Free Fatty Acid Receptors (FFAR), are widely studied within human medicine as drug targets for metabolic disorders. To combat metabolic disorders prevalent in dairy cows during the transition period, which co-occur with negative energy balance and changes to lipid and glucose metabolism, it may be helpful to identify locations and roles of FFAR and other members of the GPCR family in bovine tissues.ResultsQuantitative RT-PCR (qPCR) of subcutaneous adipose, liver, and PMNL samples during the transition period (-10, +7, and +20 or +30 d) were used for expression profiling of medium- (MCFA) and long-chain fatty acid (LCFA) receptors GPR120 and GPR40, MCFA receptor GPR84, and niacin receptor HCAR2/3. Adipose samples were obtained from cows with either high (HI; BCS ≥ 3.75) or low (LO; BCS ≤ 3.25) body condition score (BCS) to examine whether FFAR expression is correlated with this indicator of health and body reserves. Supplementation of rumen-protected methionine (MET), which may improve immune function and production postpartum, was also compared with unsupplemented control (CON) cows for liver and blood polymorphonuclear leukocytes (PMNL) samples. In adipose tissue, GPR84 and GPR120 were differentially expressed over time, while GPR40 was not expressed; in PMNL, GPR40 was differentially expressed over time and between MET vs. CON, GPR84 expression differed only between dietary groups, and GPR120 was not expressed; in liver, GPCR were either not expressed or barely detectable.ConclusionsThe data indicate that there is likely not a direct role in liver for the selected GPCR during the transition period, but they do play variable roles in adipose and PMN. In future, these receptors may prove useful targets and/or markers for peripartal metabolism and immunity.

Methionine and Choline Supply during the Periparturient Period Alter Plasma Amino Acid and One-Carbon Metabolism Profiles to Various Extents: Potential Role in Hepatic Metabolism and Antioxidant Status.

The objective of this study was to profile plasma amino acids (AA) and derivatives of their metabolism during the periparturient period in response to supplemental rumen-protected methionine (MET) or rumen-protected choline (CHOL). Forty cows were fed from −21 through 30 days around parturition in a 2 × 2 factorial design a diet containing MET or CHOL. MET supply led to greater circulating methionine and proportion of methionine in the essential AA pool, total AA, and total sulfur-containing compounds. Lysine in total AA also was greater in these cows, indicating a better overall AA profile. Sulfur-containing compounds (cystathionine, cystine, homocystine, and taurine) were greater in MET-fed cows, indicating an enriched sulfur-containing compound pool due to enhanced transsulfuration activity. Circulating essential AA and total AA concentrations were greater in cows supplied MET due to greater lysine, arginine, tryptophan, threonine, proline, asparagine, alanine, and citrulline. In contrast, CHOL supply had no effect on essential AA or total AA, and only tryptophan and cystine were greater. Plasma 3-methylhistidine concentration was lower in response to CHOL supply, suggesting less tissue protein mobilization in these cows. Overall, the data revealed that enhanced periparturient supply of MET has positive effects on plasma AA profiles and overall antioxidant status.

1629 Endocannabinoid network and proopiomelanocortin gene expression in peripartal bovine liver in response to rumen-protected methionine supplementation

1629 Endocannabinoid network and proopiomelanocortin gene expression in peripartal bovine liver in response to rumen-protected methionine supplementation

0759 Rumen-protected methyl donors during the transition period: Circulating plasma amino acids in response to supplemental rumen-protected methionine or choline

0759 Rumen-protected methyl donors during the transition period: Circulating plasma amino acids in response to supplemental rumen-protected methionine or choline

Effect of long-term rumen-protected methionine supplementation on performance of Shami goats and growth performance of their kids

An experiment was conducted to study the effect of supplementing the diets of Shami goats with rumen-protected methionine (RPM) in late pregnancy (last 60 days) and early lactation on milk production, composition, fatty acid profile in the first 60 days of lactation, and on growth performance of their kids. Three groups were used (25 goats/group) in a completely randomised design. Groups were a control with no supplements (0RPM) or supplemented with either 2.5 (2.5RPM), or 5.0 g/head.day RPM (5RPM). Birth and weaning weights for kids were recorded and analysed. Milk production was measured and sampled biweekly from each dam and analysed for milk composition. Results showed that RPM had no effect on birth and weaning weights, or average daily gain of Shami kids. However, milk to gain ratio of kids was significantly (P &amp;lt; 0.05) improved. Milk production increased significantly (P &amp;lt; 0.05) in 5RPM group compared with 0RPM and 2.5RPM groups. Milk protein content was the highest (P &amp;lt; 0.05) in the 2.5RPM group, followed by the 5RPM and 0RPM groups, whereas milk fat content was not different among the groups. Meanwhile, yields of both components were significantly (P &amp;lt; 0.05) higher in milk of treated groups. Casein and energy-corrected milk were increased (P &amp;lt; 0.05) with treatment. No effect on dry-matter intake was recorded, while feed to milk ratio was better (P &amp;lt; 0.05) for RPM groups than for 0RPM group. Milk fatty acids composition did not show significant changes after RPM treatment. In conclusion, results showed that supplementing RPM to Shami goats in late pregnancy did not affect birthweight or growth of suckling kids but improved milk production and milk protein, although no clear dose response to RPM was detected.

Maternal rumen-protected methionine supplementation and its effect on blood and liver biomarkers of energy metabolism, inflammation, and oxidative stress in neonatal Holstein calves

In nonruminants, nutrition during pregnancy can program offspring development, metabolism, and health in later life. Rumen-protected Met (RPM) supplementation during the prepartum period improves liver function and immune response in dairy cows. Our aim was to investigate the effects of RPM during late pregnancy on blood biomarkers (23 targets) and the liver transcriptome (24 genes) in neonatal calves from cows fed RPM at 0.08% of diet dry matter/d (MET) for the last 21 d before calving or controls (CON). Blood (n=12 calves per diet) was collected at birth before receiving colostrum (baseline), 24 h after receiving colostrum, 14, 28, and 50 d (post-weaning) of age. Liver was sampled (n=8 calves per diet) via biopsy on d 4, 14, 28, and 50 of age. Growth and health were not affected by maternal diet. The MET calves had greater overall plasma insulin concentration and lower glucose and ratios of glucose-to-insulin and fatty acids-to-insulin, indicating greater systemic insulin sensitivity. Lower concentration of reactive oxygen metabolites at 14 d of age along with a tendency for lower overall concentration of ceruloplasmin in MET calves indicated a lesser degree of stress. Greater expression on d 4 of fructose-bisphosphatase 1 (FBP1), phosphoenolpyruvate carboxykinase 1 (PCK1), and the facilitated bidirectional glucose transporter SLC2A2 in MET calves indicated alterations in gluconeogenesis and glucose uptake and release. The data agree with the greater expression of the glucocorticoid receptor (GR). Greater expression on d 4 of the insulin receptor (INSR) and insulin-responsive serine/threonine-protein kinase (AKT2) in MET calves indicated alterations in insulin signaling. In that context, the similar expression of sterol regulatory element-binding transcription factor 1 (SREBF1) in CON and MET during the preweaning period followed by the marked upregulation regardless of diet after weaning (d 50) support the idea of changes in hepatic insulin sensitivity during early postnatal life. Expression of carnitine palmitoyltransferase 1A (CPT1A) was overall greater and acyl-CoA oxidase 1 (ACOX1) was lower in MET calves, indicating alterations in fatty acid oxidation. Except forkhead box O1 (FOXO1), all genes changed in expression over time. Transcriptome results indicated that calves from MET-supplemented cows underwent a faster maturation of gluconeogenesis and fatty acid oxidation in the liver, which would be advantageous for adapting to the metabolic demands of extrauterine life.

Peripartal rumen-protected methionine supplementation to higher energy diets elicits positive effects on blood neutrophil gene networks, performance and liver lipid content in dairy cows.

BackgroundMain objectives were to determine to what extent Smartamine M (SM) supplementation to a prepartal higher-energy diet could alter neutrophil (PMN) and liver tissue immunometabolic biomarkers, and whether those responses were comparable to those in cows fed a prepartal lower-energy diet (CON).ResultsTwenty-eight multiparous Holstein cows were fed CON (NEL = 1.24 Mcal/kg DM) during d −50 to d −22 relative to calving. From d −21 to calving, cows were randomly assigned to a higher-energy diet (OVE, n = 9; NEL = 1.54 Mcal/kg DM), OVE plus SM (OVE + SM, n = 10; SM = 0.07 % of DM) or remained on CON (n = 9). All cows received the same basal lactation diet (NEL = 1.75 Mcal/kg DM). Supplementation of SM (OVE + SM) continued until 30 d postpartum. Liver biopsies were harvested at d −10, 7, and 21 relative to parturition. Blood PMN isolated at −10, 3, and 21 d relative to calving was used to evaluate gene expression. As expected, OVE increased liver lipid content postpartum; however, cows fed OVE + SM or CON had lower concentrations than OVE. Compared with OVE, cows in CON and OVE + SM had greater DMI postpartum and milk production. Furthermore, cows fed OVE + SM had the greatest milk protein and fat percentage and lowest milk SCC despite having intermediate PMN phagocytic capacity. Adaptations in PMN gene expression in OVE + SM cows associated with the lower SCC were gradual increases from −10 to 21 d in genes that facilitate migration into inflammatory sites (SELL, ITGAM), enzymes essential for reducing reactive oxygen metabolites (SOD1, SOD2), and a transcription factor(s) required for controlling PMN development (RXRA). The greater expression of TLR4 on d 3, key for activation of innate immunity due to inflammation, in OVE compared with CON cows suggests a more pronounced inflammatory state. Feeding OVE + SM dampened the upregulation of TLR4, despite the fact that these cows had similar expression of the pro-inflammatory genes NFKB1 and TNF as OVE. Cows in CON had lower overall expression of these inflammation-related genes and GSR, which generates reduced glutathione, an important cellular antioxidant.ConclusionsAlthough CON cows appeared to have a less stressful transition into lactation, SM supplementation was effective in alleviating negative effects of energy-overfeeding. As such, SM was beneficial in terms of production and appeared to boost the response of PMN in a way that improved overall cow health.Electronic supplementary materialThe online version of this article (doi:10.1186/s40104-016-0077-9) contains supplementary material, which is available to authorized users.

Effects of rumen-protected methionine and choline supplementation on the preimplantation embryo in Holstein cows

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.

Effect of pre-partum supplementation of rumen protected methionine plus lysine and choline on the performance of crossbred cows

The study was conducted to determine rumen escape potential (REP) of commercial rumen protected methionine (RPM), lysine (RPL) and choline (RPC) products and to examine the effect of supplementing RPM plus RPL and RPC on the performance of preparturient crossbred cows. Crossbred cows (27; 2nd to 4th lactation) were randomly assigned to 3 groups of 9 animals each, on the basis of most probable production ability (MPPA). Cows in control group (T1), were fed basal diet as per NRC (2001). The cows in second group (T2), were supplemented with 5 g RPM and 20 g RPL, and those in third group (T3), were fed 54 g RPC per cow/day for 40 days pre-partum. In sacco study revealed that REP of RPM, RPL and RPC was 75.20, 54.97 and 72.89%, respectively. Supplementation of RPM plus RPL and RPC resulted in higher body condition score (BCS) in crossbred cows on the day of parturition. Nutrient intake was similar among the groups, except that duodenal supply of methionine and lysine was higher in T2 group. Plasma triglycerides, VLDL (very low density lipoproteins) and phosphatidylcholine levels were higher in T2 and T3 groups while vitamin E level was higher in T3 group, on the day of parturition, indicating better immune status. It was concluded that fortification of ration with RPM plus RPL and RPC resulted in improved body condition score, duodenal supply of methionine and lysine and better immune status in cows during prepartum period.