RESUMO
Study objectives were to evaluate the effects of feeding rumen-protected Met (RPM) in pre- and postpartum total mixed rations (TMR) on health disorders and the interactions of health disorders with lactation and reproductive performance. Multiparous Holstein cows [470; 235 cows at University of Wisconsin (UW) and 235 cows at Cornell University (CU)] were enrolled at approximately 4 wk before parturition and housed in close-up dry cow (n = 6) and replicated lactation pens (n = 16). Pens were randomly assigned to treatment diets (pre- and postpartum, respectively): (1) control (CON): basal diet = 2.30% and 2.09% Met as % of metabolizable protein (MP) (UW) or 2.22% and 2.19% Met as % of MP (CU); (2) RPM: basal diet fed with RPM with 2.83% and 2.58% Met (Smartamine M, Adisseo Inc.; 12 g prepartum and 27 g postpartum), as % of MP (UW) or 2.85% and 2.65% Met (Smartamine M; 13 g prepartum and 28 g postpartum), as % of MP (CU). Total serum Ca was evaluated at the time of parturition and on d 3 ± 1 postpartum. Daily rumination was monitored from 7 d before parturition until 28 d postpartum. Health disorders were recorded during the experimental period until the time of first pregnancy diagnosis (32 d after timed artificial insemination; 112 ± 3 d in milk). Uterine health was evaluated on d 35 ± 3 postpartum. Time to pregnancy and herd exit were evaluated up to 350 d in milk. Treatment had no effect on the incidence of most health disorders and did not alter daily rumination. Cows fed RPM had reduced subclinical hypocalcemia (13.6 vs. 22%; UW only) on day of parturition relative to CON. Percentage of cows culled (13.1 vs. 19.3%) and hazard of herd exit due to culling [hazard ratio = 0.65, 95% confidence interval (CI): 0.42-1.02] tended to be reduced for cows fed RPM compared with CON. Moreover, cows fed RPM had greater milk protein concentration and protein yield overall, although retrospective analysis indicated that RPM only significantly increased protein yield in the group of cows with one or more health disorders (1.47 vs. 1.40 kg/d), not in cows without health disorders (1.49 vs. 1.46 kg/d) compared with CON. Overall, treatment had no effect on pregnancy per timed artificial insemination; however, among cows with health disorders, those fed RPM had reduced time to pregnancy compared with CON (hazard ratio = 0.71, 95% CI: 0.53-0.96). Thus, except for subclinical hypocalcemia on the day of parturition, feeding RPM in pre- and postpartum TMR did not reduce the incidence of health disorders, but our retrospective analysis indicated that it lessened the negative effects of health disorders on milk protein production and time to pregnancy.
Assuntos
Doenças dos Bovinos , Hipocalcemia , Gravidez , Feminino , Bovinos , Animais , Metionina/metabolismo , Rúmen/metabolismo , Hipocalcemia/veterinária , Estudos Retrospectivos , Período Pós-Parto , Reprodução , Lactação , Dieta/veterinária , Proteínas do Leite/análise , Racemetionina/metabolismo , Suplementos Nutricionais , Doenças dos Bovinos/metabolismoRESUMO
Our objective is to evaluate the effects of feeding rumen-protected Met (RPM) throughout the transition period and early lactation on the lipid profile of the preimplantation embryos and the endometrial tissue of Holstein cows. Treatments consisted of feeding a total mixed ration with top-dressed RPM (Smartamine® M, Adisseo, Alpharetta, GA, United States; MET; n = 11; RPM at a rate of 0.08% of DM: Lys:Met = 2.8:1) or not (CON; n = 9, Lys:Met = 3.5:1). Endometrial biopsies were performed at 15, 30, and 73 days in milk (DIM). Prior to the endometrial biopsy at 73 DIM, preimplantation embryos were harvested via flushing. Endometrial lipid profiles were analyzed using multiple reaction monitoring-profiling and lipid profiles of embryos were acquired using matrix assisted laser desorption/ionization mass spectrometry. Relative intensities levels were used for principal component analysis. Embryos from cows in MET had greater concentration of polyunsaturated lipids than embryos from cows in CON. The endometrial tissue samples from cows in MET had lesser concentrations of unsaturated and monounsaturated lipids at 15 DIM, and greater concentration of saturated, unsaturated (specifically diacylglycerol), and monounsaturated (primarily ceramides) lipids at 30 DIM than the endometrial tissue samples from cows in CON. In conclusion, feeding RPM during the transition period and early lactation altered specific lipid classes and lipid unsaturation level of preimplantation embryos and endometrial tissue.
RESUMO
The first objective was to investigate the effects of feeding rumen-protected methionine (RPM) during a heat stress (HS) challenge on abundance and phosphorylation of mechanistic target of rapamycin (mTOR)-related signaling proteins in mammary gland. The second objective was to investigate how HS and RPM may modulate the response of mammary gland explants to an inflammatory challenge using lipopolysaccharide (LPS). Thirty-two multiparous, lactating Holstein cows (184 ± 59 DIM) were randomly assigned to 1 of 2 environmental treatment groups, and 1 of 2 dietary treatments [TMR with RPM (Smartamine M; Adisseo Inc.; 0.105% DM as top dress) or TMR without RPM (CON)] in a crossover design. There were two periods with two phases per period. In phase 1 (9 d), all cows were in thermoneutral conditions (TN) and fed ad libitum. During phase 2 (9 d), group 1 (n = 16) cows were exposed to HS using electric heat blankets, whereas group 2 cows (n = 16) remained in TN but were pair-fed to HS counterparts to control for DMI decreases associated with HS. After a washout period (14 d), the study was repeated (period 2). Environmental treatments were inverted in period 2 (sequence), whereas dietary treatments remained the same. Mammary tissue was harvested via biopsy at the end of both periods. Tissue was used for protein abundance analysis and also for incubation with 0 or 3 µg/mL of LPS for 2 h and subsequently used for mRNA abundance. Data were analyzed using PROC MIXED in SAS. Analysis of protein abundance data included the effects of diet, environment and their interaction, and period and sequence to account for the crossover design. The explant data model also included the effect of LPS and its interaction with environment and diet. Abundance of phosphorylated mTOR and ratio of phosphorylated eukaryotic translation elongation factor 2 (p-EEF2) to total EEF2 in non-challenged tissue was greater with RPM supplementation (P = 0.04 for both) and in both cases tended to be greater with HS (P = 0.08 for both). Regardless of RPM supplementation, incubation with LPS upregulated mRNA abundance of IL8, IL6, IL1B, CXCL2, TNF, NFKB1, and TLR2 (P < 0.05). An environment × LPS interaction was observed for NFKB1 (P = 0.03); abundance was greater in LPS-treated explants from non-HS compared with HS cows. Abundance of CXCL2, NFKB1, NOS2, NOS1, and SOD2 was lower with HS (P < 0.05). Although LPS did not alter mRNA abundance of the antioxidant transcription factor NFE2L2 (P = 0.59), explants from HS cows had lower abundance of NFE2L2 (P < 0.001) and CUL3 (P = 0.04). Overall, RPM supplementation may alter mTOR activation in mammary tissue. Additionally, although HS reduced explant immune and antioxidant responses, RPM did not attenuate the inflammatory response induced by LPS in vitro.
Heat stress (HS) is an environmental issue worldwide and occurs when animals experience a heat load that exceeds their thermoregulatory capacity. Milk protein synthesis and overall production often decrease when cows are exposed to HS conditions, in part due to lower feed intake and a limit in the mammary supply of amino acids. Increasing post-ruminal supply of methionine to late-lactation cows upregulated abundance of p-mTOR in mammary tissue, providing a link with the greater milk protein production. Exposure of cows to a HS challenge also increased abundance of p-mTOR, but did not alter milk protein suggesting this response might have been associated with synthesis of other proteins. Further work at a translational level is needed to understand potential mechanisms whereby methionine may modulate mammary metabolism during periods of HS.
Assuntos
Doenças dos Bovinos , Transtornos de Estresse por Calor , Animais , Antioxidantes/metabolismo , Bovinos , Doenças dos Bovinos/metabolismo , Dieta/veterinária , Suplementos Nutricionais , Feminino , Transtornos de Estresse por Calor/metabolismo , Transtornos de Estresse por Calor/veterinária , Resposta ao Choque Térmico , Lactação , Lipopolissacarídeos/metabolismo , Metionina/farmacologia , Leite/metabolismo , RNA Mensageiro/metabolismo , Rúmen/metabolismo , Serina-Treonina Quinases TOR/metabolismoRESUMO
Multiparous, lactating Holstein cows (n = 32) were randomly assigned to one of two dietary treatments [TMR with rumen-protected Met (RPM) or TMR without RPM (CON)], and within each dietary treatment group cows were randomly assigned to one of two environmental treatment groups in a split-plot crossover design. In phase 1 (9 d), all cows were fed ad libitum and in thermoneutral conditions (TN). In phase 2 (9 d), group 1 (n = 16) was exposed to a heat stress (HS) challenge (HSC). Group 2 cows (n = 16) were pair-fed (PFTN) to HSC counterparts and remained in TN. 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, while dietary treatments remained the same for each cow. During phase 1, cows in RPM had greater plasma Met concentration compared with cows in CON (59 and 30 µM, respectively; P < 0.001). Cows in PFTN had a greater decrease (P < 0.05) in plasma insulin than cows in HSC at 4 h (-2.7 µIU/mL vs. -0.7 µIU/mL) and 8 h (-7.7 µIU/mL vs. -0.4 µIU/mL) during phase 2. Compared with cows in PFTN, cows in HSC had an increase (P < 0.05) in plasma serum amyloid A (-59 µg/mL vs. +58 µg/mL), serum haptoglobin (-3 µg/mL vs. +33 µg/mL), plasma lipopolysaccharide binding protein (-0.27 and +0.11 µg/mL), and plasma interleukin-1ß (-1.9 and +3.9 pg/mL) during phase 2. In conclusion, HSC elicited immunometabolic alterations; however, there were limited effects of RPM on cows in HSC.
Assuntos
Doenças dos Bovinos , Transtornos de Estresse por Calor , Animais , Bovinos , Dieta/veterinária , Suplementos Nutricionais , Feminino , Transtornos de Estresse por Calor/veterinária , Resposta ao Choque Térmico , Lactação , Metionina , Leite , RúmenRESUMO
Our primary objective was to evaluate the effect of feeding rumen-protected Met (RPM) in the pre- and postpartum total mixed ration (TMR) on pregnancy per artificial insemination (AI) and pregnancy loss in multiparous Holstein cows. We also evaluated multiple secondary reproductive physiological outcomes before and after AI, including uterine health, ovarian cyclicity, response to synchronization of ovulation, and markers of embryo development and size. A total of 470 multiparous Holstein cows [235 at the University of Wisconsin (UW) and 235 at Cornell University (CU)] were used for this experiment. Experimental treatment diets were applied at the pen level (2 and 4 close-up pens at CU and UW, respectively, and 12 and 6 postfresh pens at CU and UW, respectively); thus, pen was the experimental unit, and cow was the observational unit. Cows were enrolled and randomly assigned to be fed the experimental treatment diets at approximately 4 wk before parturition until 67 d of gestation [147 d in milk (DIM)] after their first service. Close-up dry cow and replicated lactation pens were randomly assigned to treatment diets: RPM, prepartum = 2.83% (UW) and 2.85% (CU), postpartum = 2.58% (UW) and 2.65% (CU); and control (CON), prepartum = 2.30% (UW) and 2.22% (CU), postpartum = 2.09% (UW) and 2.19% (CU; Met as percentage of metabolizable protein). Vaginal discharge and uterine cytology (percentage of polymorphonuclear leucocytes) were evaluated at 35 ± 3 DIM. Cows received timed AI (TAI) at 80 ± 3 DIM after synchronization of ovulation with the Double-Ovsynch protocol. Ovarian cyclicity status, response to synchronization of ovulation, and luteal function were determined by measuring circulating concentrations of progesterone at 35 and 49 ± 3 DIM, 48 and 24 h before TAI, and 8, 18, 22, 25, and 29 d after TAI. Interferon-stimulated gene expression in white blood cells were compared on 18 d after TAI (CU only) and pregnancy-specific protein B concentrations at 22, 25, 29, 32, and 67 d after TAI. Pregnancy status was determined using pregnancy-specific protein B at 25 and 29 d after TAI, and by transrectal ultrasonography at 32, 39, and 67 d after TAI. Embryo and amniotic vesicle size were determined at 32 and 39 d after TAI. Pregnancy per AI (25 d: 64.7 vs. 64.0%, 32 d: 54.3 vs. 55.1% for CON and RPM, respectively) and pregnancy loss (25 to 67 d: 22.6 vs. 19.2% for CON and RPM, respectively) for synchronized cows did not differ. The proportion of cows with purulent vaginal discharge (CON = 7.7 vs. RPM = 4.6%) and cytological endometritis (CON = 20.8 vs. RPM = 23.6%) did not differ. Cyclicity status, ovarian responses to the synchronization protocol, and synchronization rate also did not differ. In addition, fold change for interferon-stimulated genes, concentrations of pregnancy-specific protein B, and embryo size were not affected by treatments. In conclusion, feeding RPM in the pre- and postpartum TMR at the amounts used in this experiment did not affect uterine health, cyclicity, embryo development, or reproductive efficiency in dairy cows.
Assuntos
Sincronização do Estro , Rúmen , Animais , Bovinos , Dinoprosta , Feminino , Hormônio Liberador de Gonadotropina , Inseminação Artificial/veterinária , Lactação , Metionina , Período Pós-Parto , Gravidez , ProgesteronaRESUMO
Objectives were to evaluate the effect of feeding rumen-protected methionine (RPM) in pre- and postpartum total mix ration (TMR) on lactation performance and plasma AA concentrations in dairy cows. A total of 470 multiparous Holstein cows [235 cows at University of Wisconsin (UW) and 235 cows at Cornell University (CU)] were enrolled approximately 4 wk before parturition, housed in close-up dry cow and replicated lactation pens. Pens were randomly assigned to treatment diets (pre- and postpartum, respectively): UW control (CON) diet = 2.30 and 2.09% of Met as percentage of metabolizable protein (MP) and RPM diet = 2.83 and 2.58% of Met as MP; CU CON = 2.22 and 2.19% of Met as percentage of MP, and CU RPM = 2.85 and 2.65% of Met as percentage of MP. Treatments were evaluated until 112 ± 3 d in milk (DIM). Milk yield was recorded daily. Milk samples were collected at wk 1 and 2 of lactation, and then every other week, and analyzed for milk composition. For lactation pens, dry matter intake (DMI) was recorded daily. Body weight and body condition score were determined from 4 ± 3 DIM and parturition until 39 ± 3 and 49 DIM, respectively. Plasma AA concentrations were evaluated within 3 h after feeding during the periparturient period [d -7 (±4), 0, 7 (±1), 14 (±1), and 21 (±1); n = 225]. In addition, plasma AA concentrations were evaluated (every 3 h for 24 h) after feeding in cows at 76 ± 8 DIM (n = 16) and within 3 h after feeding in cows at 80 ± 3 DIM (n = 72). The RPM treatment had no effect on DMI (27.9 vs. 28.0 kg/d) or milk yield (48.7 vs. 49.2 kg/d) for RPM and CON, respectively. Cows fed the RPM treatment had increased milk protein concentration (3.07 vs. 2.95%) and yield (1.48 vs. 1.43 kg/d), and milk fat concentration (3.87 vs. 3.77%), although milk fat yield did not differ. Plasma Met concentrations tended to be greater for cows fed RPM at 7 d before parturition (25.9 vs. 22.9 µM), did not differ at parturition (22.0 vs. 20.4 µM), and were increased on d 7 (31.0 vs. 21.2 µM) and remained greater with consistent concentrations until d 21 postpartum (d 14: 30.5 vs. 19.0 µM; d 21: 31.0 vs. 17.8 µM). However, feeding RPM decreased Leu, Val, Asn, and Ser (d 7, 14, and 21) and Tyr (d 14). At a later stage in lactation, plasma Met was increased for RPM cows (34.4 vs. 16.7 µM) consistently throughout the day, with no changes in other AA. Substantial variation was detected for plasma Met concentration (range: RPM = 8.9-63.3 µM; CON = 7.8-28.8 µM) among cows [coefficient of variation (CV) > 28%] and within cow during the day (CV: 10.5-27.1%). In conclusion, feeding RPM increased plasma Met concentration and improved lactation performance via increased milk protein production.
Assuntos
Metionina , Rúmen , Animais , Bovinos , Dieta/veterinária , Suplementos Nutricionais , Feminino , Lactação , Leite , Período Pós-PartoRESUMO
The objectives of this experiment were to evaluate and compare underivatized (UND) and precolumn derivatized (DER) methods for quantification of bovine plasma AA by isotope dilution ratio via liquid chromatography-electrospray ionization (ESI)-single quadrupole mass spectrometry. Linearity of the mass-to-charge ratio signal and area signal sensitivity of 12C were evaluated for each AA with 5-point standard curves (range: 1.1-500 µM). Plasma from lactating dairy cows was isolated by centrifugation and deproteinized using 1 N perchloric acid with a final concentration of 0.5 N. Deproteinized plasma was filtered and injected into a 50 × 2-mm column (Imtakt) or extracted, derivatized, and injected into a 250 × 3-mm column (EZ:faast, Phenomenex) and analyzed via liquid chromatography-ESI-single quadrupole mass spectrometry. Coefficients of variation and recovery rates were evaluated using 4 replicates of pooled plasma samples spiked with each AA at concentrations of 10, 20, and 50 µM. In addition, a subset of 24 plasma samples was used to directly compare methods using linear regression, correlation coefficient (r), concordance correlation coefficient (CCC), and Bland-Altman plot test. Both methods showed linearity within the dynamic range analyzed for all essential AA (coefficient of determination, R2 ≥ 0.995) and most other AA, although the UND samples had poor linearity (R2 ≤ 0.990) or peak resolution problems for Asp, Gly, Tyr, and Ser. Moreover, area signal sensitivity for 12C AA was greater for DER samples than for UND samples [range: 2.2× (Pro) to 309.5× (Ala)]. Both methods had recovery rates ranging from 85.7 to 119.8.0%, and none differed from 100% except Gln [20 µM (85.7%) and 50 µM (87.6%)] and Val [50 µM (119.8%)] using the UND method. The UND method had a coefficient of variation ranging from 0.9% (Val) to 7.8% (His), whereas for the DER method the range was 2.2% (Glu) to 8.8% (Asp). The highest correlation coefficient (>0.90) and CCC (>0.90) were observed for Arg, Ile, Leu, Met, Thr, Trp, Val, and Gln, with the Bland-Altman plot test showing minimal mean bias for these AA. Lowest values were observed for His (r = 0.46; CCC = 0.45), Lys (r = 0.76; CCC = 0.75), Ala (r = 0.83; CCC = 0.73), and Glu (r = 0.65; CCC = 0.42). The UND method showed linearity, precision, and accurate recovery rates for most AA, with most essential AA having comparable values between the 2 methods. However, the DER method had greater 12C AA area signal sensitivity, linearity, and recovery rates.
RESUMO
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, ß-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.
Assuntos
Bovinos/crescimento & desenvolvimento , Dieta/veterinária , Metionina/administração & dosagem , Rúmen/metabolismo , Ácido 3-Hidroxibutírico/sangue , Aminoácidos/sangue , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Nitrogênio da Ureia Sanguínea , Peso Corporal , Butiratos/administração & dosagem , Butiratos/metabolismo , Bovinos/sangue , Doenças dos Bovinos/metabolismo , Suplementos Nutricionais , Feminino , Fermentação/efeitos dos fármacos , Masculino , Metionina/metabolismo , Leite , DesmameRESUMO
The objectives of this study were to evaluate the effects of daily top-dressing (individually feeding on the top of the total mixed ration) with rumen-protected methionine (RPM) from 30 ± 3 until 126 ± 3 Days in milk on productive and reproductive performance in lactating dairy cows. A total of 309 lactating dairy Holstein cows (138 primiparous and 171 multiparous) were randomly assigned to treatment diets containing either RPM (21.2 g of RPM + 38.8 g of dried distillers grain; 2.34% Methionine [Met] of metabolizable protein [MP]) or Control (CON; 60 g of dried distillers grain; 1.87% Met of MP). Plasma amino acids were evaluated at the time of artificial insemination (AI) and near pregnancy diagnosis. Milk production and milk composition were evaluated monthly. Pregnancy was diagnosed on Day 28 (by Pregnancy-specific protein B [PSPB]), 32, 47, and 61 (by ultrasound) and sizes of embryonic and amniotic vesicle were determined by ultrasound on Day 33 after AI. Feeding RPM increased plasma Met at 6, 9, 12, and 18 hours after top-dressing with a peak at 12 hours (52.4 vs 26.0 µM; P < 0.001) and returned to basal by 24 hours. Cows fed RPM had a small increase in milk protein percentage (3.08 vs 3.00%; P = 0.04) with no differences on milk yield and milk protein yield. Additionally, in multiparous cows, RPM feeding increased milk protein (3.03 vs 2.95%; P = 0.05) and fat (3.45 vs 3.14%; P = 0.01) percentages, although no effects were observed in primiparous cows. In multiparous cows fed RPM, pregnancy loss was lower between Days 28 to 61 (19.6 [10/51] vs. 6.1% [3/49]; P = 0.03) or between Days 32 to 61 (8.9 [4/45] vs. 0 [0/0] %; P = 0.03), although, there was no effect of treatment on pregnancy loss in primiparous cows. Consistent with data on pregnancy loss, RPM feeding increased embryonic abdominal diameter (P = 0.01) and volume (P = 0.009) and amniotic vesicle volume (P = 0.04) on Day 33 of pregnancy in multiparous cows but had no effect on embryonic size in primiparous cows. Thus, the increase in plasma Met concentrations after feeding RPM was sufficient to produce a small increase in milk protein percentage and to improve embryonic size and pregnancy maintenance in multiparous cows. Further studies are needed to confirm these responses and understand the biological mechanisms that underlie these responses as well as the timing and concentrations of circulating Met that are needed to produce this effect.
Assuntos
Ração Animal , Indústria de Laticínios , Metionina/administração & dosagem , Rúmen/metabolismo , Aminoácidos/sangue , Animais , Bovinos , Feminino , Metionina/sangue , Metionina/metabolismo , Leite/metabolismo , Gravidez , ReproduçãoRESUMO
BACKGROUND: Feeding higher-energy prepartum is a common practice in the dairy industry. However, recent data underscore how it could reduce performance, deepen negative energy balance, and augment inflammation and oxidative stress in fresh cows. We tested the effectiveness of rumen-protected methionine in preventing the negative effect of feeding a higher-energy prepartum. Multiparous Holstein cows were fed a control lower-energy diet (CON, 1.24 Mcal/kg DM; high-straw) during the whole dry period (~50 d), or were switched to a higher-energy (OVE, 1.54 Mcal/kg DM), or OVE plus Smartamine M (OVE + SM; Adisseo NA) during the last 21 d before calving. Afterwards cows received the same lactation diet (1.75 Mcal/kg DM). Smartamine M was top-dressed on the OVE diet (0.07% of DM) from -21 through 30 d in milk (DIM). Liver samples were obtained via percutaneous biopsy at -10, 7 and 21 DIM. Expression of genes associated with energy and lipid metabolism, hepatokines, methionine cycle, antioxidant capacity and inflammation was measured. RESULTS: Postpartal dry matter intake, milk yield, and energy-corrected milk were higher in CON and OVE + SM compared with OVE. Furthermore, milk protein and fat percentages were greater in OVE + SM compared with CON and OVE. Expression of the gluconeogenic gene PCK1 and the lipid-metabolism transcription regulator PPARA was again greater with CON and OVE + SM compared with OVE. Expression of the lipoprotein synthesis enzyme MTTP was lower in OVE + SM than CON or OVE. Similarly, the hepatokine FGF21, which correlates with severity of negative energy balance, was increased postpartum only in OVE compared to the other two groups. These results indicate greater liver metabolism and functions to support a greater production in OVE + SM. At 7 DIM, the enzyme GSR involved in the synthesis of glutathione tended to be upregulated in OVE than CON-fed cows, suggesting a greater antioxidant demand in overfed cows. Feeding OVE + SM resulted in lower similar expression of GSR compared with CON. Expression of the methionine cycle enzymes SAHH and MTR, both of which help synthesize methionine endogenously, was greater prepartum in OVE + SM compared with both CON and OVE, and at 7 DIM for CON and OVE + SM compared with OVE, suggesting greater Met availability. It is noteworthy that DNMT3A, which utilizes S-adenosylmethionine generated in the methionine cycle, was greater in OVE and OVE + SM indicating higher-energy diets might enhance DNA methylation, thus, Met utilization. CONCLUSIONS: Data indicate that supplemental Smartamine M was able to compensate for the negative effect of prepartal energy-overfeeding by alleviating the demand for intracellular antioxidants, thus, contributing to the increase in production. Moreover Smartamine M improved hepatic lipid and glucose metabolism, leading to greater liver function and better overall health.
RESUMO
BACKGROUND: Compared with choline, Met enhances milk yield and feed intake, and elicits a better immuno-metabolic status in periparturient cows. It is unknown whether hepatic activity and transcription of betaine-homocysteine methyltransferase (BHMT), 5-methyltetrahydrofolate-homocysteine methyltransferase (MTR), and cystathionine ß-synthase (CBS) are responsive to Met and choline supply. OBJECTIVE: This study sought to characterize hepatic BHMT, MTR, and CBS transcription and activity in response to Met and choline supplementation. METHODS: Forty multiparous cows were used in a 2 × 2 factorial design from -21 d through 30 d around parturition to assess effects of dietary rumen-protected Met (0% or 0.08% dry matter basis) or rumen-protected choline (0 or 60 g · cow-1 · d-1). Liver tissue obtained on days -10, 7, 20, and 30 was used for analyses. RESULTS: Met-supplemented cows had greater methionine adenosyltransferase 1A (MAT1A) (0.38 compared with 0.27; SEM = 0.05; P = 0.02) and phosphatidylethanolamine methyltransferase (PEMT) (0.74 compared with 0.58; SEM = 0.08; P = 0.05) expression. Greater S-adenosylhomocysteine hydrolase (SAHH) (0.93 compared with 0.74; SEM = 0.05; P = 0.01) and CBS (1.16 compared with 1.02; SEM = 0.07; P = 0.04), as well as lower MTR activity (23.4 compared with 29.7 nmol product · h-1 · mg protein-1; SEM = 2.9; P = 0.04), also were detected in Met- but not choline-supplemented cows. Although BHMT and MTR expression and BHMT enzyme activity did not change (P > 0.05), MTR enzyme activity was lower in choline-supplemented cows (23.5 compared with 29.6 nmol product · h-1 · mg protein-1; SEM = 2.9; P = 0.05). CONCLUSIONS: These findings indicate that greater synthesis of phosphatidylcholine and antioxidants contribute to the better performance and immuno-metabolic status in Met-supplemented cows. Failure to generate a comparable amount of endogenous Met from choline could be one reason that choline-fed cows fail to achieve comparable performance and health benefits during the periparturient period.
Assuntos
5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase/metabolismo , Betaína-Homocisteína S-Metiltransferase/metabolismo , Bovinos/fisiologia , Colina/administração & dosagem , Cistationina beta-Sintase/metabolismo , Metionina/administração & dosagem , 5-Metiltetra-Hidrofolato-Homocisteína S-Metiltransferase/genética , Ração Animal/análise , Fenômenos Fisiológicos da Nutrição Animal , Animais , Betaína-Homocisteína S-Metiltransferase/genética , Cistationina beta-Sintase/genética , Dieta/veterinária , Feminino , Regulação Enzimológica da Expressão Gênica , Fígado/enzimologia , Fígado/metabolismo , Período PeripartoRESUMO
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 21 days before calving to 30 days 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 60 g/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 60 g/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 30 d after calving for the presence of secretion using the Metricheck® device. On 15, 30, and 72 d 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.
RESUMO
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.
Assuntos
Aminoácidos/sangue , Fenômenos Fisiológicos da Nutrição Animal , Carbono/metabolismo , Colina/administração & dosagem , Metionina/administração & dosagem , Aminoácidos Essenciais/sangue , Ração Animal/análise , Animais , Antioxidantes/metabolismo , Bovinos , Colina/sangue , Cistationina/sangue , Cistina/sangue , Dieta/veterinária , Suplementos Nutricionais , Feminino , Homocistina/sangue , Fígado/metabolismo , Metionina/sangue , Metilistidinas/sangue , Parto , Gravidez , Prenhez , Rúmen/metabolismo , Taurina/sangue , Triptofano/sangueRESUMO
BACKGROUND: Main 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). RESULTS: Twenty-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. CONCLUSIONS: Although 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.
RESUMO
Maternal nutrition exclusively during the periconceptional period can induce remarkable effects on both oocyte maturation and early embryo development, which in turn can have lifelong consequences. The objective of this study was to evaluate the effect of maternal methionine supplementation on the transcriptome of bovine preimplantation embryos. Holstein cows were randomly assigned to one of two treatments differing in level of dietary methionine (1.89 Met vs. 2.43 Met % of metabolizable protein) from calving until embryo flushing. High quality preimplantation embryos from individual cows were pooled and then analyzed by RNA sequencing. Remarkably, a subtle difference in methionine supplementation in maternal diet was sufficient to cause significant changes in the transcriptome of the embryos. A total of 276 genes out of 10,662 showed differential expression between treatments (FDR <0.10). Interestingly, several of the most significant genes are related to embryonic development (e.g., VIM, IFI6, BCL2A1, and TBX15) and immune response (e.g., NKG7, TYROBP, SLAMF7, LCP1, and BLA-DQB). Likewise, gene set enrichment analysis revealed that several Gene Ontology terms, InterPro entries, and KEGG pathways were enriched (FDR <0.05) with differentially expressed genes involved in embryo development and immune system. The expression of most genes was decreased by maternal methionine supplementation, consistent with reduced transcription of genes with increased methylation of specific genes by increased methionine. Overall, our findings provide evidence that supplementing methionine to dams prior to conception and during the preimplantation period can modulate gene expression in bovine blastocysts. The ramifications of the observed gene expression changes for subsequent development of the pregnancy and physiology of the offspring warrant further investigation in future studies.
Assuntos
Blastocisto , Suplementos Nutricionais , Metionina/administração & dosagem , Transcriptoma , Animais , Sequência de Bases , Bovinos , Primers do DNA , Feminino , Regulação da Expressão Gênica no Desenvolvimento , GravidezRESUMO
The early embryo's nutritional environment plays an important role in establishing its developmental potential. However, little is known about the specific nutrient requirements of the embryo. The objective of the present study was to determine requirements of the in vitro produced bovine embryo for the essential amino acid methionine. In addition to serving as a precursor for polypeptides, methionine plays roles in regulation of translation, DNA methylation, and antioxidant balance. In the first experiment, embryos were cultured in potassium simplex optimized medium - bovine embryo modification 2 containing 0, 35, 50, 100, 200 or 400 µmol/l L-methionine for 8 days. There was no effect of methionine concentration on cleavage rate. The percent of oocytes that developed to blastocyst was lower for embryos without methionine at Day 7 and 8 than other groups but was similar for embryos cultured with 35-400 µmol/l. Neither total cell number, allocation of cells to trophectoderm or inner cell mass, or frequency of apoptosis was affected by methionine concentration. In the second experiment, embryos were cultured with 0, 7, 14, 21, 28 or 35 µmol/l methionine. There was no effect of methionine concentration on cleavage rate. The percent of oocytes that developed to blastocyst was lower for embryos without methionine at Day 7 and 8 but was not different between embryos cultured with 7-35 µmol/l methionine. However, the proportion of blastocysts that were expanded, hatching or hatched on Day 7 was reduced at lower concentrations of methionine (7 and 14). DNA methylation of blastocyst nuclei was unaffected by methionine concentration but intracellular glutathione content was higher for embryos cultured without methionine. In conclusion, the methionine requirement for preimplantation development is between 14 and 21 µmol/l. These concentrations are lower or similar to those found in the reproductive tract and suggest that methionine deficiency is not a common cause of embryonic mortality.