Rumen-protected Methionine Research Articles

Effects of rumen-protected methionine or methionine analogs in starter on plasma metabolites, growth, and efficiency of Holstein calves from 14 to 91 d of age

During weaning, methionine (Met) supply decreases as liquid feed intake is reduced and ruminal function is developing. During this transition, the calf starter should both promote ruminal development and provide adequate nutrients post-ruminally. In mature ruminants, rumen-protected Met (RPM) and the Met analogs, 2-hydroxy-4-(methylthio)-butanoic acid (HMTBa) and HMTBa isopropyl ester (HMBi), are used to increase Met supply, stimulate ruminal fermentation, or exert both effects, respectively. To evaluate the effects of these forms of Met on calf performance during development of ruminal function, 74 Holstein calves were raised until 91 d of age, in 2 enrollment periods. Calves were individually housed from birth and, at 14 d of age, balanced for sex and randomly assigned to receive a starter with no added Met (CTRL, n = 20) or one supplemented with RPM (Smartamine M, Adisseo USA Inc., Alpharetta, GA; n = 16), HMTBa (RumenSmart, Adisseo; n = 19), or HMBi (MetaSmart, Adisseo; n = 19). Milk replacer [28% crude protein (CP), 15% fat] was offered up to 1.6 kg of dry matter (DM)/d and fed 3 times daily. Weaning was facilitated from d 49 to 63. The 4 starters (25% CP, 2.5 Mcal of metabolizable energy/kg of DM) were offered ad libitum, and supplement inclusion was set to provide an additional 0.16% DM of Met equivalents, and equal amounts of HMTBa within the analogs. Body weight and stature were measured, and blood was collected and analyzed for plasma urea nitrogen, β-hydroxybutyrate, and free AA on a weekly basis. Supplementation of RPM and HMBi increased free plasma Met, but no differences in growth or feed efficiency compared with calves fed the CTRL starter could be attributed to the additional Met supply alone. The addition of HMBi in the starter increased feed intake and body weight during the last weeks of the experiment. On the contrary, HMTBa failed to increase plasma Met and depressed intake and growth after weaning, likely because the level included in the diet was too high and intake was greater than previous studies, exacerbating the level of HMTBa ingested. No differences were observed in stature, feed efficiency, or non-AA plasma measurements among groups. These results demonstrate that RPM and HMBi are effective sources of metabolizable Met; however, Met was apparently not limiting calves fed the basal diet in this study. The increased feed intake observed with the inclusion of HMBi in the starter during the weaning and early postweaning period might be mediated by its metabolism in the rumen, and further research is needed to determine the mechanisms involved.

Preconditioning beef cattle for long-duration transportation stress with rumen-protected methionine supplementation: A nutrigenetics study.

In general, beef cattle long-distance transportation from cow-calf operations to feedlots or from feedlots to abattoirs is a common situation in the beef industry. The aim of this study was to determine the effect of rumen-protected methionine (RPM) supplementation on a proposed gene network for muscle fatigue, creatine synthesis (CKM), and reactive oxygen species (ROS) metabolism after a transportation simulation in a test track. Angus × Simmental heifers (n = 18) were stratified by body weight (408 ± 64 kg; BW) and randomly assigned to dietary treatments: 1) control diet (CTRL) or 2) control diet + 8 gr/hd/day of top-dressed rumen-protected methionine (RPM). After an adaptation period to Calan gates, animals received the mentioned dietary treatment consisting of Bermuda hay ad libitum and a soy hulls and corn gluten feed based supplement. After 45 days of supplementation, animals were loaded onto a trailer and transported for 22 hours (long-term transportation). Longissimus muscle biopsies, BW and blood samples were obtained on day 0 (Baseline), 43 (Pre-transport; PRET), and 46 (Post-transport; POST). Heifers’ average daily gain did not differ between baseline and PRET. Control heifer’s shrink was 10% of BW while RPM heifers shrink was 8%. Serum cortisol decreased, and glucose and creatine kinase levels increased after transportation, but no differences were observed between treatments. Messenger RNA was extracted from skeletal muscle tissue and gene expression analysis was performed by RT-qPCR. Results showed that AHCY and DNMT3A (DNA methylation), SSPN (Sarcoglycan complex), and SOD2 (Oxidative Stress-ROS) were upregulated in CTRL between baseline and PRET and, decreased between pre and POST while they remained constant for RPM. Furthermore, CKM was not affected by treatments. In conclusion, RPM supplementation may affect ROS production and enhance DNA hypermethylation, after a long-term transportation.

Bioavailability of rumen-protected methionine, lysine and histidine assessed by fecal amino acid excretion

The objective of this experiment was to assess a new method to determine the bioavailability (BA) of rumen-protected (RP)His, RPLys and RPMet products by determining rumen escape (RE) and fecal excretion of undigested AA from RPAA. Eight lactating Holstein cows (79 ± 21 days in milk, 53.0 ± 7.0 kg/d milk yield), four of which rumen-cannulated, were used in a replicated 4 × 4 Latin square design experiment with four, 26-day experimental periods and a preceding 22-day background (BG) period. Four combinations of nine commercial and experimental RPAA (HisA, HisB, LysA, LysB, LysC, MetA, MetB, MetC, MetD) were fed daily to supply 20, 25, and 35 g/day of digestible (d)His, dLys and dMet, respectively. The following treatment combinations were used: (1) HisALysAMetA, (2) HisBLysBMetB, (3) LysCMetC and (4) LysCMetD. Spot sampling of feces was performed during the BG period to establish basal levels of AA in feces. Total fecal collection and blood sampling were performed during the last three days of each experimental period. Rumen escape fraction of each RPAA product was determined in situ and was greater for HisA (0.90) than for HisB (0.64), ranged from 0.33 to 0.85 (SEM = 0.017) for RPLys and from 0.53 to 0.95 (SEM = 0.017) for RPMet. Apparent post-ruminal digestibility of AA from RPAA was calculated as [(RE of AA, g/day – BG corrected fecal AA output, g/day) ÷ RE of AA, g/day]. Digestibility was similar between the two RPHis products (0.85 and 0.90). Apparent post-ruminal digestibility of Lys from RPLys products varied from 0.30 to 0.79 and digestibility for RPMet varied from 0.85 to 0.96. Bioavailability was calculated as: (RE, g/g × AA digestibility, g/g) × 100 and was greater for HisA compared with HisB (76.1 and 57.9 % respectively), varied from 9.63 (LysC) to 67.0 % (LysA) for the RPLys and was lowest for MetC (49.3 %) and greatest for MetD (91.9 %) among the RPMet products. Plasma His concentration was greater for HisA than for HisB and plasma Met concentration was greater for MetD compared with the other three RPMet products, reflecting estimated BA of the RPAA products. In contrast, plasma Lys concentration did not differ among RPLys products. This study showed that a method using fecal AA output in combination with RE of RPAA can reveal differences in BA between RPAA products. This is a first step in establishing the fecal AA excretion technique for determination of BA of RPAA and warrants further validation.

Effects of rumen-protected methionine supplementation on growth performance, nitrogen balance, carcass characteristics, and meat quality of lambs fed diets containing buckwheat straw

This study was conducted to determine the effects of rumen-protected methionine (RPM) supplementation on growth performance, nitrogen (N) balance, carcass traits, and meat quality of lambs fed diets containing buckwheat straw. Twenty male Tan lambs from Ningxia were housed in individual pens and were randomly assigned to five dietary treatments that were supplemented with RPM at levels of 0 (control), 1.5 (T1), 3.0 (T2), 4.5 (T3), and 6.0 g d−1 (T4). Dry matter intake was higher in T4 than control (P < 0.05). The feed:gain ratio generally reduced with increasing dietary RPM, but the lowest ratio was found in T3 (P < 0.05). Dry matter digestibility and N retention were highest in T3 (P < 0.05) while N excretion was the lowest. Carcass fat weight and grid reference tissue depth increased while drip loss decreased by supplemental RPM, and significant differences were found between T4 and control (P < 0.05). However, pH, color, cooking loss, and shear force were not affected. The results from this study demonstrated that dietary RPM supplementation may be able to improve the feed efficiency, N utilization, and meat quality of lambs; however, the optimal dose of RPM needs to be explored further.

The impact of rumen-protected amino acids on the expression of key- genes involved in the innate immunity of dairy sheep.

Rumen protected amino acids inclusion in ewes’ diets has been proposed to enhance their innate immunity. The objective of this work was to determine the impact of dietary supplementation with rumen-protected methionine or lysine, as well as with a combination of these amino acids in two different ratios, on the expression of selected key-genes (NLRs, MyD88, TRIF, MAPK-1, IRF-3, JunD, TRAF-3, IRF-5, IL-1α, IL-10, IKK-α, STAT-3 and HO-1). Thus, sixty Chios dairy ewes (Ovis aries) were assigned to one of the following five dietary treatments (12 animals/ treatment): A: basal diet consist of concentrates, wheat straw and alfalfa hay (control group); B: basal diet +6.0 g/head rumen-protected methionine; C: basal diet + 5.0 g/head rumen-protected lysine; D: basal diet +6.0 g/head rumen-protected methionine + 5.0 g/head rumen-protected lysine and E: basal diet +12.0 g/head rumen-protected methionine + 5.0 g/head rumen-protected lysine. The results revealed a significant downregulation of relative transcript level of the IL-1α gene in the neutrophils of C and in monocytes of D ewes compared with the control. Significantly lower mRNA transcript accumulation was also observed for the MyD88 gene in the neutrophils of ewes fed with lysine only (C). The mRNA relative expression levels of JunD gene were highly induced in the monocytes, while those of IL-10 and HO-1 genes were declined in the neutrophils of ewes fed with the C and D diets compared with the control. Lower transcript levels of STAT-3 gene were observed in the neutrophils of ewes fed with either C or with E diets in comparison with the control. In conclusion, our results suggest that the dietary supplementation of ewes with rumen-protected amino acids, down regulate the expression of some genes involved in the pro-inflammatory signalling.

Body Temperature and Reproductive Performance of Beef Heifers Supplemented With Rumen-Protected Methionine

This study aimed to assess the reproductive performance and body temperature of beef heifers supplemented with rumen-protected methionine (RPM) in an extensive grazing system and mated during the summer. One hundred 5/8 Angus 3/8 Brahman heifers were homogeneously divided into the control group (n=48, CG) supplemented with mineral salt ad libitum and methionine group (n=52, MG) who received the same supplementation plus 4 g of methionine per 100 g of mineral salt. The groups were fixed-time artificial inseminated (FTAI) 45 days after starting the supplementation. On day 0 of the FTAI protocol, ten animals per group received data-logger thermometers attached in progesterone implants recording the body temperature of the animals every 30 minutes. Pregnancy diagnosis was performed on days 30 and 60 after the artificial insemination to determine the rates of conception, pregnancy, and pregnancy loss. At the beginning of the supplementation, both groups had similar weights (MG was 304.6 kg and CG 304.4 kg), while on days 0 and 30 about the FTAI protocol, the MG was higher weight (365.9 vs. 346.7 and 384.8 vs. 371.5 Kg). During the FTAI protocol, the MG had lower body temperatures (38.77 vs. 38.99 °C) and a tendency to have larger dominant follicles than the CG (11.76 mm x 10.8 mm). Although it did not impact reproductive performance, the observed higher weight, dominant follicle sizes, and lower body temperatures during FTAI protocols show the potential benefits of methionine supplementation in beef heifers mated during the summer.

Effects of Dietary Rumen-Protected Betaine on Lactation Performance and Serum Metabolites of Mid-lactation Holstein Dairy Cows

This experiment was conducted to investigate the effects of dietary rumen-protected betaine (RPB) supplementation, as partial replacement for methionine, on the lactation performance of mid-lactation dairy cows. A total of 36 Holstein dairy cows were randomly assigned to three groups [control, 20 g/day RPB, or 15 g/day rumen-protected methionine (RPM)]. The experiment was conducted over 9 weeks, with the first week for adaptation. Blood metabolites were analyzed with metabolomics in the control and RPB groups. The results revealed that the milk yield and milk protein content were higher in cows fed RPB and RPM compared to those in the control group. Concentrations of nine metabolites differed between cows in the RPB and control groups. These metabolites were mainly concentrated in six pathways, such as arginine synthesis and proline degradation and cyanoamino acid synthesis. This study revealed that RPB can spare methionine and improve lactation performance of dairy cows fed with diets moderately deficient in methionine.

Production and nitrogen metabolism in lactating dairy cows fed finely ground field pea plus soybean meal or canola meal with or without rumen-protected methionine supplementation

We showed previously that dairy cows fed [diet dry matter (DM) basis] 25% finely ground field pea (GFP) plus rumen-protected (RP)-Met and RP-Lys had greater milk true protein yield and plasma Met concentration, but lower plasma His, compared with those fed GFP without rumen-protected AA supplementation. The goal of the present study was to investigate the effects of diets containing soybean meal (SBM) or canola meal (CM) with or without a source of RP-Met on production, nutrient digestibility, and N metabolism in cows fed 25% GFP. Sixteen mid-lactation Holstein cows were used in a replicated 4 × 4 Latin square with a 2 × 2 factorial arrangement of treatments (21-d periods). Cows were fed (DM basis) 35% corn silage, 14% grass-legume haylage, 25% GFP, and 11% SBM or 13.5% CM with or without 0.095% RP-Met supplementation as Smartamine M (Adisseo USA Inc., Alpharetta, GA). Protein source effects were observed for most production variables; cows fed diets containing CM plus GFP had greater DM intake, yields of milk and milk fat and true protein, and milk N efficiency than those offered SBM plus GFP. Feeding CM plus GFP also reduced the concentrations of milk urea N and plasma urea N, and the urinary excretion of NH3 N and urea N, suggesting improved N use efficiency. Moreover, replacing SBM with CM increased the concentrations of all essential AA (except Arg) in plasma collected from the coccygeal blood vessels. A protein source × RP-Met interaction was observed for the concentration of His in coccygeal plasma, with circulating His decreasing only when RP-Met was supplemented to the diet containing SBM plus GFP. Based on the arteriovenous difference method, Lys was the first limiting AA overall, with Met being the first limiting AA in diets that did not receive RP-Met. Apparent total-tract digestibilities of DM, organic matter, N, and acid detergent fiber increased with feeding SBM plus GFP versus CM plus GFP. Most variables were not affected by RP-Met supplementation, except plasma Met concentration, which increased by 63%. Collectively, our results indicate that CM appears to be a better companion rumen-degradable protein source to GFP than SBM, due to improved yields of milk and milk protein and N use efficiency.

Effects of rumen-protected methionine on lactation performance and physiological variables during a heat stress challenge in lactating Holstein cows

Milk yield, content, and composition are altered by heat stress. Thirty-two multiparous, lactating Holstein cows [balanced by days in milk (mean ± standard deviation; 184 ± 59); body surface area (5.84 ± 0.34 m2)] were randomly assigned to 1 of 2 dietary treatments [total mixed ration with rumen-protected Met (RPM; Smartamine M; Adisseo Inc., Antony, France; 1.05 g of RPM/kg of dry matter intake) or total mixed ration without RPM (CON)], and within each dietary treatment group cows were randomly assigned to 1 of 2 environmental treatment groups in a split-plot crossover design. The study was divided into 2 periods with 2 phases per period. In phase 1 (9 d), all cows were in thermoneutral conditions and fed ad libitum. In phase 2 (9 d), group 1 (n = 16) was exposed to a heat stress challenge (HSC) using electric heat blankets. Group 2 (n = 16) remained in thermoneutral conditions but was pair-fed (PFTN) to HSC counterparts. After a 21-d washout period, the study was repeated (period 2) and the environmental treatments were inverted relative to treatments from phase 2 of period 1, whereas dietary treatments (RPM or CON) remained the same for each cow. Cows were milked 3× per day and samples were taken on d 1, 5, and 9 of each phase. Vaginal temperature was measured every 10 min, rectal temperature and skin temperature were measured 3× per day, and respiration rate and heart rate were recorded once per day. Cow activity was measured using an accelerometer. Paired difference values were calculated for each cow for each period based on the difference between phase 1 baseline means and phase 2 values for each variable. Cows in HSC had a greater increase in vaginal temperature and respiration rate (+0.2°C and +13.7 breaths/min, respectively) compared with cows in PFTN (0.0°C and -1.6 breaths/min, respectively). Cows in PFTN had a greater decrease in dry matter intake and milk yield (-3.9 and -2.6 kg/d, respectively) compared with cows in HSC (-3.2 and -0.9 kg/d, respectively). Cows in CON had a greater decrease in milk protein concentration for PFTN (-0.10 percentage units) and HSC (-0.06 percentage units) compared with cows in RPM for PFTN (0.00 percentage units) and HSC (-0.02 percentage units). Cows in CON for HSC had greater decrease in milk fat concentration compared with cows in RPM for HSC (-0.10 and +0.12 percentage units, respectively). In conclusion, HSC altered physiological and production parameters of cows. Additionally, RPM helped maintain milk protein and fat concentration during HSC, whereas dry matter intake, milk yield, and feed efficiencies were not affected by RPM.

Dietary supplementation of rumen-protected methionine decreases the nitrous oxide emissions of urine of beef cattle through decreasing urinary excretions of nitrogen and urea.

Two consecutive trials were carried out to study the effects of dietary supplementation of rumen-protected methionine (RPM) on nutrient digestibility, nitrogen (N) metabolism (Trial 1), and consequently the nitrous oxide (N2 O) emissions from urine in beef cattle (Trial 2). Eight 24-month-old castrated Simmental bulls with liveweights of 494 ± 28 kg, and four levels of dietary supplementation of RPM at 0, 10, 20, and 30 g head-1 d-1 , were allocated in a replicated 4 × 4 Latin square for Trial 1 and the N2 O emissions from the urine samples collected in Trial 1 were measured using a static incubation technique in Trial 2. Supplementation of RPM at 0, 10, 20, and 30 g head-1 d-1 to a basal ration deficient in methionine (Met) did not affect the apparent digestibility of dry matter, organic matter, neutral detergent fiber, or acid detergent fiber (P > 0.05), but decreased the urinary excretions of total N (P < 0.05) and urea (P < 0.001), increased the ratio of N retention / digested N (P < 0.05) in beef cattle, and decreased the estimated cattle urine N2 O-N emissions by 19.5%, 23.4%, and 32.6%, respectively (P < 0.001). Supplementation of RPM to Met-deficient rations was effective in improving the utilization rate of dietary N and decreasing the N2 O emissions from urine in beef cattle. © 2019 Society of Chemical Industry.

PSXIV-4 Fetal programming effect of rumen-protected methionine on Angus-Simental heifer offspring’s performance and adipogenesis, oxidative stress and DNA methylation gene expression regulation

Abstract An extensive body of literature reported positive effects of Rumen-Protected Methionine (RPM) on dairy cattle; however, its effects are unknown on beef cattle. The objective of this study was to assess the effect of RPM supplementation on Angus-Simmental heifer’s offspring during late gestation, lactation, and after weaning (~65d). Ninety days prior to calving, 22 Angus-Simmental heifers were stratified by body weight (454 ± 85 kg; BW) and randomly assigned to dietary treatments: 1) Control diet (CTRL), and 2) Control diet + RPM (RPM). Dams had access to Bermuda hay ad libitum, and a soy hull:corn gluten based supplement. After weaning, calves were allocated by dietary treatments based on their dam’s treatment. Calves received Bermuda hay ad libitum and early-wean feed supplement after weaning. Offspring’s BW and Longissimus dorsi (LM) muscle samples were obtained on days 0, 25, 50, 100 after weaning. Messenger RNA was extracted from LM for RT-qPCR analysis. There was a tendency for RPM calves to be heavier at birth compared to control calves (P = 0.07). Although, no differences in BW were detected at weaning and after weaning (P = 0.20). Calves average daily gain between birth and weaning was greater for RPM calves as compared to CON (P = 0.02) suggesting a greater milk production, with no differences after weaning (P = 0.53). RT-qPCR results showed a treatment × time interaction for PPARG, LPL, CEBPG, DNMT1, SOD2 and NOS3 (P &amp;lt; 0.05). Furthermore, PPARG had a treatment and a time effect, which was upregulated in RPM calves (P &amp;lt; 0.01) from 50 to 100 days after weaning. In conclusion, dam’s RPM supplementation during late gestation increased offspring’s BW when inside the womb and probably their milk production. Offspring’s RPM supplementation after weaning potentially stimulates an early differentiation of adipocytes in LM samples, detected by PPARG upregulation.

197 Gene-regulation control of muscle fatigue during transportation with rumen-protected methionine supplementation

Abstract Different participants generally located distant among them compose the U.S. beef production chain, therefore shipping is required for cattle at least once in their lifespan. The aim of this study was to determine the effect of Rumen-Protected Methionine (RPM) supplementation on muscle fatigue gene network, creatine synthesis (CKM), and Reactive Oxygen Species (ROS) metabolism after a transportation simulation in a 1.7-miles oval test track. Angus heifers (n = 18) were stratified by body weight (408 ± 64 kg; BW) and randomly assigned to dietary treatments: 1) control diet (CTRL), and 2) control diet + rumen-protected methionine (RPM). After a successful adaptation period to Calan gates, animals received a common diet of Bermuda hay ad libitum and a soy hulls and corn gluten feed based supplement. After 45 days under supplementation, animals were loaded onto a 32 × 7 ft. trailer and transported for 22 hours. Skeletal muscle biopsies, BW and blood samples were obtained on day 0 (Baseline), 43 (Pre-transport, “PRET”), and 45 (Post-transport, “POST”). Heifer’s average daily gain did not differ between Baseline and PRET (P = 0.41). Control heifer’s shrink was 8% of BW while RPM heifers shrink was 10% (P = 0.29). Cortisol level decreased after transportation, but no differences were observed between treatments (P = 0.94). Messenger RNA was extracted from skeletal muscle tissue and gene expression analysis was performed by RT-qPCR. Results showed that AHCY (Creatine synthesis pathway), SSPN (Sarcoglycan complex), DNMT3A (DNA Methylation), and SOD2 (Oxidative Stress-ROS) were upregulated (P &amp;lt; 0.05) in CTRL between baseline and PRET and, decreased between PRET and POST (P &amp;lt; 0.05) while they remained constant for RPM. Furthermore, CKM was not affected by treatments (P = 0.11). In conclusion, muscle fatigue related genes were not affected by RPM. Although, RPM could affect ROS production, after a long-term transportation.

398 Effects of protein level and supplemental methionine in late-gestation on colostrum quality and passive immunity transfer in beef cattle

Abstract The objective of this study was to evaluate the influence of metabolizable protein (MP) level and supplemental rumen-protected methionine (RPM) on colostrum quality and passive immunity transfer from beef cows fed during late-gestation. One hundred and forty-seven pregnant Angus crossbred cows and heifers were randomly assigned to one of six dietary treatments based on a 3 x 2 factorial arrangement. Diets included feeding to 90%, 100% and 110% MP requirements (NRC, 2016), offered with (without) 9 g/d of RPM. Cattle were individually fed a partially mixed ration (60% haylage/40% straw for cows; 70% haylage/30% straw for heifers) with supplements top-dressed daily for approximately 56 d prior to calving. Immediately after calving, a colostrum sample was collected from each dam for analysis of milk components, total protein, and IgG concentrations. Two d post-calving, serum samples were collected from each calf for analysis of total protein and IgG concentrations. A refractometer was used to estimate total protein as a preliminary estimate of the transfer of maternal IgG and total IgG concentrations were later determined using Radial Immunodiffusion (RID). Data were analyzed as a completely randomized block design using Proc GLIMMIX in SAS. Level of MP, RPM, and their interaction did not influence concentrations of fat, protein, BHB or SCC in colostrum (P ≥ 0.12). Supplementation of RPM decreased concentrations of MUN (45.92 vs. 54.51 ±3.17 mg/dl; P = 0.04), but did not change IgG (14.66 vs. 15.90 ±0.87 g/dl; P = 0.29). While MP level during gestation did not affect total protein or IgG concentrations in calf serum (P ≥ 0.64), providing supplemental RPM decreased total protein (5.79 vs. 6.33 ±0.13 g/dl; P = 0.002) and IgG concentrations in calf serum (3.44 vs. 4.30 ±0.25 g/dl; P = 0.01). These results suggest that providing supplemental methionine for beef cows during gestation may alter colostrum quality and decrease serum protein and IgG in offspring.

394 Exploring the relationships between cow-calf management systems as affected by protein/methionine nutrition during gestation on the performance of cow-calf pairs prior to weaning

Abstract A three-way factorial arrangement was used to assess impacts of protein/methionine nutrition during gestation and two lactational management systems on performance of cow-calf pairs pre-weaning. 140 crossbred beef cows were managed in drylot and randomly assigned to one of six nutritional treatments during the third trimester of gestation. Cows were fed to meet 110% (HP), 100% (MP), or 90% (LP) of metabolizable protein requirements for late-gestation (NRC, 2016), with(without) rumen-protected methionine (RPM) for 8 weeks prior to parturition. Post-partum, cow-calf pairs were managed in drylot (DL) and fed ad libitum, a ration formulated to meet lactational nutrient requirements, or rotationally grazed on pasture (PAS). Cow body weights (BW) and body condition scores (BCS) and calf BW were recorded at regular intervals. All data were analyzed using PROC GLIMMIX (SAS v9.4). HP cows were heaviest at calving (P = 0.02), but calf birthweight was unaffected by gestational nutrition (P &amp;gt; 0.19). Gestational nutrition did not impact cow conception rate (CR), or calf BW and ADG (P &amp;gt; 0.17). Lactational management system (PAS, DL) did not affect cow BCS or CR (P &amp;gt; 0.13), but impacted cow and calf BW and ADG. PAS calves were heavier, had greater ADG, and heavier weaning BW than DL calves (P &amp;lt; 0.02). This came at the expense of BW loss throughout lactation and lower ADG for PAS cows, versus BW gains and greater ADG for DL cows (P &amp;lt; 0.0001). An interaction between protein level and RPM supplementation was present for cow BW at calving and weaning, and BW change throughout lactation (P &amp;lt; 0.04), with tendencies for interactions for cow ADG and BCS (P &amp;lt; 0.09). Overall, prepartum protein/methionine supplementation did not affect calf performance prior to weaning, but may have affected cow performance during lactation. Rotationally grazing cow-calf pairs on pasture improved pre-weaning calf gains, but reduced cow gains throughout lactation.

PSIV-B-25 Late-Breaking: Maternal supply of polyunsaturated fatty acids and methionine during late- gestation alters amino acid transporters and global DNA methylation in the lamb small intestine

Abstract The intestine plays a crucial role in nutrient digestion and absorption, and its function is critical for animal growth and health. However, the effect of maternal nutrition on offspring’s intestine is not well documented. The objective of this study was to evaluate the effects of maternal supply of polyunsaturated fatty acids, methionine or both during late-gestation on offspring’s intestine amino acids transporters and global DNA methylation. Twenty ewes were used in a randomized block design and treatments were arranged in a 2×2 factorial design. Treatments were: 1) control diet (CTR) without polyunsaturated fatty acids (PUFA) or methionine (MET) supplementation; or supplemented with 2) Ca salts of PUFA (1% DMI; Strata G113®, Virtus Nutrition); rumen-protected MET (0.1% DMI; Smartamine®, Adisseo); and 4) PUFA and MET (PUFA+MET). Treatments were fed from day 100 to 145 of gestation. On day 145, ewes were euthanized and fetuses’ intestines were sampled. Protein quantification of amino acid transporters was measured using Simple Western System and global DNA methylation was determined using a commercial kit. No interactions between PUFA×MET were observed for the variables analyzed. PUFA increased (P ≤ 0.03) global DNA methylation and protein expression of SLC7A5, SLC38A2, and SLC38A10 compared with CTR. Furthermore, PUFA tended to increase (P = 0.07) SLC38A1 protein expression compared with CTR, whereas no difference (P &amp;gt; 0.05) was observed for SLC6A9. Compared with CTR, MET increased (P ≤ 0.05) global DNA methylation and SLC38A10 protein expression, and tented to increase (P ≤ 0.10) SLC6A19 and SLC38A2. MET did not affect SLC7A5 and SLC38A1 compared with CTR. Maternal supply of PUFA and MET led to greater protein expression of amino acid transporters and global DNA methylation on offspring’ intestine; however, the effects of these nutrients were not additive. Further research is necessary to better understand the role of DNA methylation on intestine’s amino acid transporters.

76 Does supplemental protein and rumen-protected methionine improve performance and digestibility during late-gestation in beef cows?

Abstract Methionine (MET) is likely the first limiting amino acid in low-protein forages fed to beef cattle during late-gestation. Therefore, the objective of this study was to determine if supplemental protein and rumen-protected MET improve cow performance and apparent tract digestibility (TTD) during late-gestation. This study used 147 late-gestation Angus crossbred cows and heifers in a 3x2 factorial arrangement for dietary treatments. The cattle were randomly assigned to one of six diets formulated to 90, 100 or 110% of metabolizable protein (MP) requirements (NRC, 2016), with (without) 9 g/d of rumen-protected MET (MetaSmart, Adisseo Inc.). These diets were fed for approximately 8 wks before calving. All data was analyzed as a randomized block design using SAS 9.4 PROC GLIMMIX procedure. Cows fed at 90% MP requirements lost body weight (BW), while cows fed at 100% and 110% MP requirements maintained and (or) gained BW over the trial (P = 0.02). Similarly, cows fed at 90% MP requirements lost more pregnancy corrected BW than cows fed at 100% and 110% MP requirements (P = 0.01). However, supplemental MET did not affect body weight gains (P &amp;gt; 0.07). Cows and heifers fed at 90% MP requirements had reduced TTD for crude protein compared to cattle fed to 110% MP requirements (P &amp;lt; 0.001), MET supplementation did not impact TTD (P &amp;gt; 0.20). Additionally, cattle fed at 90% MP requirements had increased serum cholesterol and reduced urea concentrations compared to cows fed to 100% and 110% MP requirements (P &amp;lt; 0.0001). MET supplementation increased serum concentrations for glucose, isoleucine, leucine, lysine, serine, threonine and valine (P &amp;lt; 0.02). Calf birth weights were not significantly impacted by dietary treatment (P &amp;gt; 0.31). Feeding cows above their MP requirements may improve late-gestation performance and CP digestibility. Supplemental MET may increase amino acid utilization but did not improve beef cow performance or digestibility parameters measured in late-gestation.

PSVI-5 Rumen-protected methionine supplementation during the peripartal period reduces the expression of LGALS-1, -3 and -4 in polymorphonuclear leukocytes (PMNL) and secretion of Gal-2 and Gal-12 in plasma of Holstein cows

Abstract The objective of this study was to evaluate the effect of methionine(Met) supplementation on genes encoding Galectins (LGALS) and secretion in plasma during the periparturient period. Dairy cows are susceptible to increased incidence and severity of disease during the periparturient period. Impaired polymorphonuclear leukocyte (PMNL) functions such as phagocytosis and pathogen killing capacity contribute to immune dysfunction and disease. Galectins (GALS) are an evolutionarily conserved family of animal lectins important in disease and homeostasis. They behave like pathogen-associated molecular patterns and cell-surface receptors whose activation often results in signaling cascades to activate cells such as PMNL. Fourteen Holstein cows supplemented with either rumen-protected Met (n = 7) or no Met (CON; n = 7) were used. Diets were fed from −21 ± 2 before expected calving date and continued until 30 days postpartum. To evaluate LGALS expression and secretion of Gal, PMNL were isolated from blood and plasma was harvested at -10, +7, and +30 d relative to expected calving. The expression of LGALS1 (P = 0.09), LGALS3 (P = 0.02), and LGALS4 (P = 0.02) was lower in response to Met, but there was no effect on LGALS8, LGALS9, and LGALS12. Met supplementation reduced Gal-2 and Gal-12 secretion in plasma at -10, +7, and +30 d (P &amp;lt; 0.0005 and P &amp;lt; 0.0001 respectively). Methionine supplementation modulates the expression of GAL involved in inflammation and lipolysis in cow blood. Modulation of GAL in PMNL and plasma in response to Met may have implications for the use of dietary interventions to target gene expression in PMNL for improved function and needs further study.

Hepatic Cystathionine β-Synthase Activity Is Increased by Greater Postruminal Supply of Met during the Periparturient Period in Dairy Cows

ABSTRACTBackgroundPostruminal supply of Met during the periparturient period enhances production efficiency (feed conversion to milk) in dairy cows partly through alleviation of oxidant and inflammatory status. Whether alterations in hepatic 1-carbon metabolism (major contributor of antioxidants) and/or energy metabolism contribute to these beneficial effects is unknown.ObjectivesTo investigate alterations in hepatic 1-carbon and energy metabolism and associations with plasma amino acids (AAs) and production efficiency in response to enhanced postruminal supply of Met.MethodsHolstein cows (n = 30 per group) were fed during the last 28 d of pregnancy a control diet (CON) or the control plus ethylcellulose rumen-protected Met (MET; 0.9 g/kg of dry matter intake). Plasma (n = 15 per group) and liver tissue (n = 10 per group) were collected throughout the periparturient period to evaluate AA profiles, activity of the tricarboxylic acid cycle, and 1-carbon metabolism via mRNA abundance, enzyme activity, and targeted metabolomics.ResultsCows in the MET group had greater overall (27%, P = 0.027) plasma Met concentrations, but had similar total plasma AA concentrations. Although mRNA abundance of 1-carbon metabolism enzymes did not differ, hepatic activity of cystathionine β-synthase (CBS) (51.2 compared with 44.4 mmol/h/mg protein; P = 0.032) and concentration (19%, P = 0.048) of the cellular antioxidant glutathione were greater overall in the MET group. mRNA abundance of aconitase 2 and fumarate hydratase was greater overall (P = 0.049), and phosphoenolpyruvate carboxykinase 1 tended (P = 0.093) to be greater overall in cows fed MET. There was a tendency (P ≤ 0.093) for greater overall hepatic concentrations of malic acid, α-ketoglutaric acid, and isocitric acid in cows fed MET.ConclusionsGreater activity of CBS in response to enhanced postruminal supply of Met likely contributes to alleviating oxidant status by increasing concentrations of glutathione. Hence, transsulfuration plays an important role in the observed improvements in production efficiency of dairy cows during the periparturient period.

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.

Humoral immune response in beef heifers supplemented with mineral salt with or without the addition of rumen-protected methionine

The aim of this study was to evaluate the humoral immune response in beef heifers supplemented with mineral supplementation with or without the addition of rumen-protected methionine. Forty-eight Brangus nulliparous heifers were distributed into four experimental groups with three replications each: control group without supplementation and without vaccination (CG01), control group without supplementation and with vaccination (CG02), treatment group with mineral supplementation and vaccination (TG01), and treatment group with mineral supplementation added with protected methionine and vaccination (TG02). The animals were maintained under native pasture with access to water ad libitum and the supplementation was available in high-consumption covered troughs. A supplementation period of 60 days prior to vaccinations was adopted until the first dose of a monovalent experimental vaccine inactivated for BoHV-5 was applied as a method of stimulating the immune response to evaluate the supplementation effects. After a 21-day interval, blood samples were collected to evaluate the humoral response and the second vaccine booster dose was applied following the 21-day interval for new blood samples in order to evaluate the immune response against the two-vaccination protocol. From the beginning of the experiment, the animals were weighed on the days ?60, ?10, 0, 21, and 42 in relation to the vaccine protocol. The experimental groups did not differ for body weight, mean daily weight gain, and body condition score after 102 days of supplementation regardless of the treatment. No animals belonging to CG01 seroconverted throughout the experiment, proving that there was no introduction of the agent (BoHV) in the studied area. When vaccinated animals were compared to the CG01 control group, statistically higher levels of neutralizing antibodies (P ? 0.0001) and IgG (P ? 0.0001) were verified 21 days after the second vaccine dose. Among the animals of the three vaccinated groups, there was no difference in seroconversion and IgG production. Therefore, no benefits of mineral supplementation or enriched with protected methionine were observed for the humoral immune response of the studied animals.