Supply of palmitic, stearic, and oleic acid changes rumen fiber digestibility and microbial composition.

The concept that fat supplementation impairs total-tract fiber digestibility in ruminants has been widely accepted over the past decades. Nevertheless, the recent interest in the dietary fatty acid profile to dairy cows enlightened the possible beneficial effect of specific fatty acids (e.g., palmitic, stearic, and oleic acids) on total-tract fiber digestibility. Because palmitic, stearic, and oleic acids are the main fatty acids present in ruminal bacterial cells, we hypothesize that the dietary supply of these fatty acids will favor their incorporation into the bacterial cell membranes, which will support the growth and enrichment of fiber-digesting bacteria in the rumen. Our objective in this experiment was to investigate how dietary supply of palmitic, stearic, and oleic acid affect fiber digestion, bacterial membrane fatty acid profile, microbial growth, and composition of the rumen bacterial community. Diets were randomly assigned to 8 single-flow continuous culture fermenters arranged in a replicated 4 × 4 Latin square with four 11-d experimental periods. Treatments were (1) a control basal diet without supplemental fatty acids (CON); (2) the control diet plus palmitic acid (PA); (3) the control diet plus stearic acid (SA); and (4) the control diet plus oleic acid (OA). All fatty acid treatments were included in the diet at 1.5% of the diet (dry matter [DM] basis). The basal diet contained 50% orchardgrass hay and 50% concentrate (DM basis) and was supplied at a rate of 60 g of DM/d in 2 equal daily offers (0800 and 1600 h). Data were analyzed using a mixed model considering treatments as fixed effect and period and fermenter as random effects. Our results indicate that PA increased in vitro fiber digestibility by 6 percentage units compared with the CON, while SA had no effect and OA decreased fiber digestibility by 8 percentage units. Oleic acid decreased protein expression of the enzymes acetyl-CoA carboxylase compared with CON and PA, while fatty acid synthase was reduced by PA, SA, and OA. We observed that PA, but not SA or OA, altered the bacterial community composition by enhancing bacterial groups responsible for fiber digestion. Although the dietary fatty acids did not affect the total lipid content and the phospholipid fraction in the bacterial cell, PA increased the flow of anteiso C13:0 and anteiso C15:0 in the phospholipidic membrane compared to the other treatments. In addition, OA increased the flow of C18:1 cis-9 and decreased C18:2 cis-9,cis-12 in the bacterial phospholipidic membranes compared to the other treatments. Palmitic acid tended to increase bacterial growth compared to other treatments, whereas SA and OA did not affect bacterial growth compared with CON. To our knowledge, this is the first research providing evidence that palmitic acid supports ruminal fiber digestion through shifts in bacterial fatty acid metabolism that result in changes in growth and abundance of fiber-degrading bacteria in the microbial community.

Alterations in skeletal muscle abundance of protein turnover, stress, and antioxidant proteins during the periparturient period in dairy cows fed ethyl-cellulose rumen-protected methionine.

Skeletal muscle turnover helps support the physiological needs of dairy cows during the transition into lactation. We evaluated effects of feeding ethyl-cellulose rumen-protected methionine (RPM) during the periparturient period on abundance of proteins associated with transport AA and glucose, protein turnover, metabolism, and antioxidant pathways in skeletal muscle. Sixty multiparous Holstein cows were used in a block design and assigned to a control or RPM diet from -28 to 60 d in milk. The RPM was fed at a rate of 0.09% or 0.10% of dry matter intake (DMI) during the prepartal and postpartal periods to achieve a target Lys:Met ratio in the metabolizable protein of ∼2.8:1. Muscle biopsies from the hind leg of 10 clinically healthy cows per diet collected at -21, 1, and 21 d relative to calving were used for western blotting of 38 target proteins. Statistical analysis was performed using the PROC MIXED statement of SAS version 9.4 (SAS Institute Inc.) with cow as random effect, whereas diet, time, and diet × time were the fixed effects. Diet × time tended to affect prepartum DMI, with RPM cows consuming 15.2 kg/d and controls 14.6 kg/d. However, diet had no effect on postpartum DMI (17.2 and 17.1 ± 0.4 kg/d for control and RPM, respectively). Milk yield during the first 30 d in milk was also not affected by diet (38.1 and 37.5 ± 1.9 kg/d for control and RPM, respectively). Diet or time did not affect the abundance of several AA transporters or the insulin-induced glucose transporter (SLC2A4). Among evaluated proteins, feeding RPM led to lower overall abundance of proteins associated with protein synthesis (phosphorylated EEF2, phosphorylated RPS6KB1), mTOR activation (RRAGA), proteasome degradation (UBA1), cellular stress responses (HSP70, phosphorylated MAPK3, phosphorylated EIF2A, ERK1/2), antioxidant response (GPX3), and de novo synthesis of phospholipids (PEMT). Regardless of diet, there was an increase in the abundance of the active form of the master regulator of protein synthesis phosphorylated MTOR and the growth-factor-induced serine/threonine kinase phosphorylated AKT1 and PIK3C3, whereas the abundance of a negative regulator of translation (phosphorylated EEF2K) decreased over time. Compared with d 1 after calving and regardless of diet, the abundance of proteins associated with endoplasmic reticulum stress (XBP1 spliced), cell growth and survival (phosphorylated MAPK3), inflammation (transcription factor p65), antioxidant responses (KEAP1), and circadian regulation (CLOCK, PER2) of oxidative metabolism was upregulated at d 21 relative to parturition. These responses coupled with the upregulation of transporters for Lys, Arg, and His (SLC7A1) and glutamate/aspartate (SLC1A3) over time were suggestive of dynamic adaptations in cellular functions. Overall, management approaches that could take advantage of this physiological plasticity may help cows make a smoother transition into lactation.

Unique adaptations in neonatal hepatic transcriptome, nutrient signaling, and one-carbon metabolism in response to feeding ethyl cellulose rumen-protected methionine during late-gestation in Holstein cows.

BACKGROUND: Methionine (Met) supply during late-pregnancy enhances fetal development in utero and leads to greater rates of growth during the neonatal period. Due to its central role in coordinating nutrient and one-carbon metabolism along with immune responses of the newborn, the liver could be a key target of the programming effects induced by dietary methyl donors such as Met. To address this hypothesis, liver biopsies from 4-day old calves (n = 6/group) born to Holstein cows fed a control or the control plus ethyl-cellulose rumen-protected Met for the last 28 days prepartum were used for DNA methylation, transcriptome, metabolome, proteome, and one-carbon metabolism enzyme activities. RESULTS: Although greater withers and hip height at birth in Met calves indicated better development in utero, there were no differences in plasma systemic physiological indicators. RNA-seq along with bioinformatics and transcription factor regulator analyses revealed broad alterations in 'Glucose metabolism', 'Lipid metabolism, 'Glutathione', and 'Immune System' metabolism due to enhanced maternal Met supply. Greater insulin sensitivity assessed via proteomics, and efficiency of transsulfuration pathway activity suggested beneficial effects on nutrient metabolism and metabolic-related stress. Maternal Met supply contributed to greater phosphatidylcholine synthesis in calf liver, with a role in very low density lipoprotein secretion as a mechanism to balance metabolic fates of fatty acids arising from the diet or adipose-depot lipolysis. Despite a lack of effect on hepatic amino acid (AA) transport, a reduction in metabolism of essential AA within the liver indicated an AA 'sparing effect' induced by maternal Met. CONCLUSIONS: Despite greater global DNA methylation, maternal Met supply resulted in distinct alterations of hepatic transcriptome, proteome, and metabolome profiles after birth. Data underscored an effect on maintenance of calf hepatic Met homeostasis, glutathione, phosphatidylcholine and taurine synthesis along with greater efficiency of nutrient metabolism and immune responses. Transcription regulators such as FOXO1, PPARG, E2F1, and CREB1 appeared central in the coordination of effects induced by maternal Met. Overall, maternal Met supply induced better immunometabolic status of the newborn liver, conferring the calf a physiologic advantage during a period of metabolic stress and suboptimal immunocompetence.

Effect of feeding a palmitic acid-enriched supplement on production responses and nitrogen metabolism of mid-lactating Holstein and Jersey cows.

This study evaluated the effect of feeding a palmitic acid-enriched supplement on production responses and nitrogen metabolism of mid-lactating Holstein and Jersey cows. Eighty mid-lactating dairy cows, 40 Holstein and 40 Jersey, were used in a randomized complete block design with a split-plot arrangement; the main plot was breed and the subplot was fatty acid treatment. Cows within each breed were assigned to 1 of 2 treatments: (1) control diet with no fat supplement or (2) control diet plus a palmitic acid-enriched supplement dosed at 1.5% of diet dry matter (PA treatment). The treatment period was 6 wk with the final 3 wk used for data and sample collection. There were no treatment × breed interactions for the variables analyzed. Compared with control, PA treatment increased milk fat yield (1.36 vs. 1.26 kg/d) and tended to increase 3.5% fat-corrected milk (35.6 vs. 34.0 kg/d) and energy-corrected milk (35.7 vs. 34.1 kg/d). There was no effect of PA treatment on dry matter intake, milk yield, milk protein yield, milk lactose yield, body condition score, body weight (BW) change, nitrogen intake, and variables related to nitrogen metabolism and excretion. Compared with Holstein cows, Jersey cows had greater dry matter intake as a percent of BW (4.90 vs. 3.37% of BW) and lower milk production (29.6 vs. 32.7 kg/d) and milk lactose yield (1.58 vs. 1.42 kg/d), but tended to have greater milk fat yield (1.36 vs. 1.26 kg/d). There was a breed effect on BW change; Holstein cows gained 0.385 kg/d during the experiment, and Jersey cows gained 0.145 kg/d. Jersey cows had lower nitrogen intake (636 vs. 694 g/d), blood urea nitrogen (12.6 vs. 13.8 mg/dL), urine total nitrogen (125 vs. 145 g/d), and urine total nitrogen as a percent of nitrogen intake (19.5 vs. 21.1%). Overall, feeding a palmitic acid-enriched supplement increased milk fat yield as well as dry matter and fiber digestibility in both Holstein and Jersey cows. The PA treatment did not have any major effects on nitrogen metabolism in both Holstein and Jersey cows. In addition, our results indicated that Jersey cows had lower urinary nitrogen excretion (g/d) than Holstein cows.

Methionine Supply During Late-Gestation Triggers Offspring Sex-Specific Divergent Changes in Metabolic and Epigenetic Signatures in Bovine Placenta.

Background: Nonruminant male and female offspring respond differently to gestational nutrition, with placenta contributing to the underlying mechanisms. However, similar data are lacking in large ruminants. Objectives: The aim of this study was to investigate the impact of methionine supply during late-gestation on metabolism and DNA methylation in placenta from cows carrying male or female calves. Methods: During the last 28 d of pregnancy, cows were individually fed a control diet (CON) or the control diet plus rumen-protected d,l-methionine (MET; 0.9 g/kg dry matter intake). Placentomes collected at term were classified according to cow dietary treatment and offspring sex as follows: Male CON (n = 7), Male MET (n = 7), Female CON (n = 8), and Female MET (n = 8). Calf growth was measured until 9 wk of age. Targeted metabolomics, RT-PCR, global DNA methylation, and activity of selected enzymes in one-carbon metabolism and transsulfuration pathways were performed. Statistics were conducted via ANOVA using MIXED models. Results: At birth, Male MET calves were heavier than Male CON calves (7.6%, P = 0.02), but body mass was similar at 9 wk of age. In contrast, compared with Female CON, Female MET calves had greater body mass at 9 wk of age (6.3%, P = 0.03). Compared with Male CON, placenta from Male MET calves had greater concentrations of tricarboxylic acid (TCA) cycle and transsulfuration intermediates (23-100%, P < 0.05), along with greater 5-methyltetrahydrofolatehomocysteine methyltransferase activity (67%, P = 0.03). Compared with Female CON, placenta from Female MET calves had greater concentrations of one-carbon metabolism intermediates (13-52%, P < 0.05). DNA methyltransferase 3A (DNMT3A) was upregulated (43%, P < 0.01) in placenta from Female MET compared with Female CON calves. Global DNA methylation was lower in placenta from Female MET compared with Female CON calves (45%, P = 0.06). Conclusions: Methionine supply affects placental metabolism, DNA methylation, and body mass of the calf in a sex-specific manner, underscoring its importance as dietary methyl-donor for pregnant cows.

Placentome Nutrient Transporters and Mammalian Target of Rapamycin Signaling Proteins Are Altered by the Methionine Supply during Late Gestation in Dairy Cows and Are Associated with Newborn Birth Weight.

Background: To our knowledge, most research demonstrating a link between maternal nutrition and both fetal growth and offspring development after birth has been performed with nonruminants. Whether such relationships exist in large ruminants is largely unknown.Objective: We aimed to investigate whether increasing the methionine supply during late pregnancy would alter uteroplacental tissue nutrient transporters and mammalian target of rapamycin (mTOR) and their relation with newborn body weight.Methods: Multiparous Holstein cows were used in a randomized complete block design experiment. During the last 28 d of pregnancy, cows were fed a control diet or the control diet plus ethylcellulose rumen-protected methionine (0.9 g/kg dry matter intake) (Mepron; Evonik Nutrition & Care GmbH) to achieve a 2.8:1 ratio of lysine to methionine in the metabolizable protein reaching the small intestine. We collected placentome samples at parturition and used them to assess mRNA and protein expression and the phosphorylation status of mTOR pathway proteins.Results: Newborn body weight was greater in the methionine group than in the control group (44.1 kg and 41.8 kg, respectively; P ≤ 0.05). Increasing the methionine supply also resulted in greater feed intake (15.8 kg/d and 14.6 kg/d), plasma methionine (11.9 µM and 15.3 µM), and plasma insulin (1.16 µg/L and 0.81 µg/L) in cows during late pregnancy. As a result, mRNA expression of genes involved in neutral amino acid transport [solute carrier (SLC) family members SLC3A2, SLC7A5, SLC38A1, and SLC38A10], glucose transport [SLC2A1, SLC2A3, and SLC2A4], and the mTOR pathway [mechanistic target of rapamycin and ribosomal protein S6 kinase B1] were upregulated (P ≤ 0.07) in methionine-supplemented cows. Among 6 proteins in the mTOR pathway, increasing the methionine supply led to greater (P ≤ 0.09) protein expression of α serine-threonine kinase (AKT), phosphorylated (p)-AKT, p-eukaryotic elongation factor 2, and the p-mTOR:mTOR ratio.Conclusion: Supplemental methionine during late gestation increases feed intake and newborn body weight in dairy cows, and this effect may be mediated by alterations in the uteroplacental transport of nondispensable and dispensable amino acids and glucose at least in part through changes in gene transcription and mTOR signaling.

Effect of sources of calcium salts of fatty acids on production, nutrient digestibility, energy balance, and carryover effects of early lactation grazing dairy cows.

The objective of our study was to investigate the effects of sources of calcium salts of fatty acids (FA) on production, nutrient digestibility, energy balance, and carryover effects of early lactation grazing dairy cows. Treatment diets were offered from 3 to 16 wk postpartum (the treatment period), in which all cows grazed elephantgrass (Pennisetum purpureum 'Cameroon') and treatments were added to a concentrate supplement. The treatments were (1) control (concentrate without supplemental fat); (2) concentrate with calcium salts of soybean FA (CSSO); and (3) concentrate with calcium salts of palm FA (CSPO). From 17 to 42 wk postpartum (the carryover period), all cows received a common diet fed as a total mixed ration. During the treatment period, CSPO increased milk yield, milk fat yield, 3.5% fat-corrected milk, energy-corrected milk, and cumulative milk yield compared with control and CSSO. Treatment CSSO increased the yield of milk but did not affect 3.5% fat-corrected milk or energy-corrected compared with control. Also, CSSO decreased milk fat yield, dry matter intake, neutral detergent fiber digestibility, and body weight and body condition loss. Compared with control, both CSSO and CSPO increased feed efficiency (3.5% fat-corrected milk:dry matter intake), and CSPO increased feed efficiency compared with CSSO. When considering energy partitioning (as % energy intake), CSPO increased energy partitioning toward milk and increased energy mobilized from body reserves compared with control and CSSO. Furthermore, CSSO tended to reduce the mobilization of energy from body reserves compared with control. In the carryover period, no differences in milk composition were observed among treatments. A treatment by time interaction was observed during the carryover period for milk yield because cows on CSPO maintained higher production compared with control and CSSO cows until 30 wk postpartum; CSSO had a lower carryover effect sustaining higher milk yield compared with control until 25 wk postpartum. In conclusion, supplementation with CSPO was an effective strategy to increase energy intake and yields of milk and milk solids and it had a greater carryover effect. Supplementation with CSSO resulted in lower mobilization of reserves and less variation in body weight and body condition throughout lactation.

Corn grain-processing method interacts with calcium salts of palm fatty acids supplementation on milk production and energy balance of early-lactation cows grazing tropical pasture.

The objective of our study was to investigate the associative effects of feeding Ca salts of palm fatty acids (FA) and corn grain-processing method on production, nutrient digestibility, energy balance, and carryover effects of early-lactation dairy cows grazing a tropical pasture. Treatment diets were offered from 3 to 16 wk postpartum (treatment period), in which all cows grazed elephant grass (Pennisetum purpureum L. Cameroon) and treatments were added to a concentrate supplement. Treatments were flint corn grain-processing method either as fine ground (FGC) or steam-flaked (SFC) associated with Ca salts of palm FA supplementation either not supplemented or supplemented (CSPO). From 17 to 40 wk postpartum (carryover period) all cows received a common diet fed as total mixed ration. During the treatment period, a tendency for an interaction between CSPO and corn grain-processing method were observed for milk yield, milk fat yield, and energy-corrected milk (ECM), as CSPO caused them to increased to a greater extent in the FGC diet compared with the SFC diet. Furthermore, a tendency for an interaction between CSPO and corn grain-processing method was observed for body weight change, because CSPO increased body weight loss in the FGC diet but not in the SFC diet. The CSPO increased milk yield, milk fat yield, 3.5% fat-corrected milk, ECM, and cumulative milk yield compared with not supplemented. Also, CSPO increased energy intake, milk energy output, and energy partitioning toward milk, whereas reduced energy was allocated to body reserves. The SFC increased milk yield, ECM, milk protein yield, milk casein yield, and cumulative milk yield, and decreased milk urea N compared with FGC. The SFC compared with FGC also increased body condition score and body weight change, and increased energy partitioning toward body reserves. During the carryover period, an interaction between CSPO and corn grain-processing method was observed for milk yield, which occurred because CSPO maintained higher milk yield in the FGC diet but not in the SFC diet. Therefore, in the carryover period, the additive effect between SFC and CSPO that occurred in the treatment period was not maintained throughout the carryover period. However, CSPO increased yields of milk fat, protein, casein as well as fat-corrected milk and ECM. In conclusion, corn grain-processing method interacts with CSPO supplementation on production responses and carryover effects of grazing cows. When CSPO was fed in the FGC diet, milk production increased to a greater extent than when fed in the SFC diet, but also caused greater mobilization of reserves at early lactation. This suggests an interaction between fat supplementation and corn grain-processing method on energy partitioning of dairy cows. Also, both supplementation with CSPO and SFC were effective strategies to increase energy intake and yields of milk and milk solids. The carryover effect on milk production was greater for CSPO supplementation than corn grain-processing method, whereas feeding SFC diets had lower mobilization of reserves and less body weight and body condition score variation throughout lactation.

Replacing soybean meal for cottonseed meal on performance of lactating dairy cows.

Cottonseed meal (CSM) is an alternative source of protein, and previous studies have been shown that it can replace soybean meal (SBM) without decrease animal performance. However, Brazilian CSM has a different chemical composition compared with the usual CSM reported in the literature. The objective of this experiment was to evaluate the effects of replacing SBM for Brazilian CSM on performance and energy balance of mid-lactating dairy cows. Forty-two Holstein cows were used in a replicate 3 × 3 Latin square design. Increasing contents of CSM (0, 15, and 30% of dry matter (DM)) were fed in diets to replace SBM. Milk yield and feed efficiency were linearly reduced with the replacement of CSM for SBM (P = 0.001). Milk fat content tended to increase quadratically (P = 0.07) with CSM addition. Replacing SBM for CSM affected milk protein content quadratically (P = 0.05). Milk urea nitrogen and plasma urea nitrogen (PUN) tended to respond quadratically (P = 0.06 and 0.10) when CSM replaced SBM to the diets. Variation in body weight (BW) also responded quadratically as CSM replaced SBM (P = 0.05). Altogether, the findings suggest better performance when cows receive SBM diet compared with the Brazilian CSM diet.

Supplementation with Ca salts of soybean oil interacts with concentrate level in grazing dairy cows: milk production and milk composition.

In this study, we investigated the associative effects of concentrate levels and Ca salts of soybean oil (CSSO) supplementation on milk production, milk composition, and milk fatty acids of mid-lactation dairy cows grazing on tropical pasture. Twenty-four Jersey × Holstein cows were used in a randomized block design and assigned to four treatments arranged in a 2 × 2 factorial design. Factors evaluated were concentrate levels (low, 3 kg/day vs. high, 7 kg/day of concentrate) and CSSO supplementation (without CSSO vs. with 250 g CSSO cow/day). All cows grazed on elephant grass (Pennisetum purpureum cv. Cameroon) and received the supplemental treatments for a 90-day period. Interactions between concentrate level and CSSO were detected for milk yield, milk yield components, energy-corrected milk (ECM) and 3.5 % fat-corrected milk (FCM). Milk yield increased when CSSO was fed in a low concentrate level, while it decreased milk production in a high concentrate level. Yields of fat, protein, lactose, 3.5 % FCM, and ECM were not affected with CSSO in the low concentrate, but reduced in the high concentrate level. CSSO increased proportions of monounsaturated milk FA, C18:2 trans-10 cis-12, and polyunsaturated FA, and reduced proportions of saturated milk FA in milk. In conclusion, feeding the high level of concentrate was an effective strategy to improve milk yield and solid production. CSSO supplementation increased milk production when fed at low concentrate level but did not affect yield of solids.

Supplementation with Ca salts of soybean oil interacts with concentrate level in grazing dairy cows: intake, ingestive behavior, and ruminal parameters.

In this study, we investigated the associative effects of concentrate levels and Ca salts of soybean oil (CSSO) supplementation on performance and ruminal parameters of mid-lactation dairy cows grazing on tropical pasture. Twenty-four Jersey × Holstein cows were used in a randomized block design and assigned to four treatments arranged in a 2 × 2 factorial design. Factors evaluated were concentrate levels (low, 3 kg/day vs. high, 7 kg/day of concentrate) and CSSO supplementation (without CSSO vs. with 250 g CSSO cow/day). All cows grazed on elephant grass (Pennisetum purpureum cv. Cameroon) and received the supplemental treatments for a 90-day period. The high concentrate level decreased forage intake and grazing time. In addition, the high concentrate level increased rumen propionate concentration and microbial synthesis and tended to decrease ammonia-N compared with low concentrate level. The addition of CSSO tended to decrease valerate, isobutyrate, isovalerate, and microbial synthesis. In conclusion, feeding CSSO for mid lactating cows grazing on tropical pasture had negative effects on rumen function. In contrast, CSSO supplementation tended to interact with concentrate level and increased energy intake when fed at low concentrate level. Feeding the high level of concentrate was an effective strategy to increase energy intake and microbial synthesis and improve N utilization.

Replacing soybean meal for wet brewer's grains or urea on the performance of lactating dairy cows.

The purpose of this research was to evaluate the partial replacement of soybean meal (SBM) for wet brewer's grains (WBG) or urea on the performance of lactating dairy cows. The second investigated whether WBG ensiled with corn kept animal performance in low- and high-producing dairy cows compared with WBG. In experiment I, 40 Holstein cows were used in 4 × 4 Latin square design. The treatments comprised WBG or urea as partial replacement for SBM, as follows: control (diet based on SBM and 1 % of urea), 10 % of WBG, 20 % of WBG, and 2 % of urea. Dry matter intake (DMI) was not affected by treatments. WBG increased milk yield linearly, but it decreased with urea. Milk fat content responded quadratically to WBG levels. Milk protein content decreased, while plasma urea nitrogen increased with high urea addition. In experiment II, 42 Holstein cows were divided into two groups according to production levels. Eighteen cows composed the group of low producing, while the high-producing group comprised 24 cows. The experimental design was a crossover with two periods of 14 days. The experimental treatments consisted of feeding WBG or WBG ensiled with ground corn. Regardless of the production level, no difference in milk yield and milk composition between treatments was observed.

Evaluation of external markers to estimate fecal excretion, intake, and digestibility in dairy cows.

The objective of this study was to compare the use of an external marker titanium dioxide (TiO2) as an alternative to chromic oxide (Cr2O3) in dairy cows. Four dairy cows were allocated in individual pens and fed concentrate supplement and Pennisetum purpureum cv. Cameroon cut daily. Fecal excretion, forage and total dry matter (DM) intakes, and digestibility were measured and estimated with TiO2 and Cr2O3. Chromic oxide overestimated and TiO2 tended to overestimate fecal excretion compared with total fecal collection. Forage and total DM intakes were overestimated by Cr2O3. The apparent DM digestibility was underestimated by Cr2O3 and TiO2. The organic matter (OM) digestibility was underestimated by both markers. There were greater mean bias, mean squared prediction error, and root of the mean squared prediction errors for all parameters estimated with Cr2O3. In conclusion, estimates using TiO2 were more precise and accurate indicating that it can replace Cr2O3 as an external marker for grazing dairy cows.

Starch levels on performance, milk composition and energy balance of lactating dairy cows.

The objective of this experiment was to evaluate the effects of starch levels in diets with the replacement of citrus pulp for corn on milk yield, milk composition, and energy balance of lactating dairy cows. Twenty-eight multiparous Holstein cows were used in seven 4 × 4 Latin squares conducted concurrently, and each experimental period consisted of 20 days (16 days for adaptation and 4 days for sampling). The experimental treatments comprised four starch levels: 15, 20, 25, and 30% in the diet. The dry matter intake increased linearly with increasing starch levels. The milk yield and 3.5% fat-corrected milk yield showed quadratic response to increasing starch levels. The milk protein content and milk total solids content responded linearly to increasing starch levels. The feed efficiency, milk lactose content, milk urea nitrogen, plasma urea nitrogen, and plasma glucose concentration were not affected by starch levels. The estimated net energy for lactation (NEL) intake increased linearly as the starch level was raised. Although the milk NEL output per kilogram of milk was not affected by starch, the milk NEL output daily responded quadratically to starch levels. In addition, the NEL in body weight gain also responded quadratically to increasing starch levels. The efficiency of energy use for milk yield and the NEL efficiency for production also responded quadratically to increasing starch levels. Diets for mid-lactating dairy cows producing around 30 kg/day of milk should be formulated to provide around 25% starch to optimize performance.

Methionine supply during mid-gestation modulates the bovine placental mTOR pathway, nutrient transporters, and offspring birth weight in a sex-specific manner.

The mechanistic target of rapamycin (mTOR) predominantly regulates the expression and activity of placental nutrient transporters. The mTOR pathway can be activated by several nutrients, including the essential amino acid methionine. Additionally, previous research in nonruminant animals suggests that mTOR is influenced in a sexually dimorphic manner. In bovine, there is limited understanding of how maternal nutrition and offspring sexual dimorphism affect the placental transfer of nutrients. Thus, we investigated the effects of increasing the supply of dietary methionine to beef cattle heifers during mid-gestation on mTOR signaling, placental nutrient transporters, and fetal growth in male and female offspring. Forty purebred Angus heifers were used in a randomized complete block design experiment. From day 90 to 180 of gestation, heifers received a basal diet with no added methionine (CON, n = 20), or the basal diet plus 8.3 g of rumen-protected methionine (MET, n = 20) per animal daily. All animals received a basal diet in the first and third trimesters of gestation. Cotyledonary tissue samples were collected at parturition and utilized to examine the mTOR pathway and nutrient transporters through protein and gene expression analysis. The offspring's body weight was measured at birth. Data were analyzed using a mixed model that included the fixed effect of treatment, offspring sex, their interactions, and the random effect of block. At day 170 of gestation, MET-supplemented heifers showed higher plasma concentrations of methionine and glutamate (P < 0.01) and lower glycine and proline levels (P ≤ 0.01) compared to the CON group. A treatment × sex interaction was observed for calf birth weight (P = 0.03). In heifers that delivered male calves, MET supplementation increased the birth weight of the calves (P < 0.01). However, the dietary treatments had no effect on the birth weight of female calves (P = 0.32). The increase in birth weight of male calves from MET-fed heifers resembles with the upregulation of placental mTOR and phosphorylated mTOR (P ≤ 0.03), as well as the amino acid transporters SLC1A5, SLC7A5, SLC38A6, and SLC38A11, and the glucose transporters SLC2A1 and SLC2A8 (P ≤ 0.05). Our findings suggest that increasing the supply of methionine to beef heifers during mid-gestation can modulate placental nutrient transport and fetal growth in a sex-dependent manner and that these effects are mediated, at least in part, by the mTOR pathway.

Roughage level and supplemental fat for newly received finishing calves: effects on growth performance, health, and physiological responses.

This experiment evaluated the effects of roughage levels and supplemental fat on intake, growth performance, health, and physiological responses of newly received finishing cattle during 58-d receiving period. A total of 72 crossbred steers (initial body weight [BW] = 200 ± 13 kg) were used in a randomized complete block design with a 2 × 2 factorial arrangement of treatments, consisting of two roughage levels (wheat hay at 30% [R30] or 60% [R60]; dry matter [DM] basis) and two levels of supplemental fat (yellow grease at 0% [-FAT; no additional fat] or 3.5% [+FAT]; DM basis). Upon arrival, calves were individually weighed, blocked by off-truck shrunk BW, and assigned to 24 soil-surfaced pens (three calves per pen). Shrunk BW was also collected on day 58 for the calculation of average daily gain (ADG). Throughout the study, calves were assessed for bovine respiratory disease (BRD). Effects of roughage level × supplemental fat interaction were only observed for diet particle size distribution and estimated physically effective neutral detergent fiber (peNDF) of diets (P ≤ 0.10). Adding fat to R60 diets tended to increase the percentage of particles retained in the 8-mm screen (P = 0.06) and the estimated peNDF (P = 0.10), but did not affect R30 diets. Dietary roughage level did not affect DM intake (DMI; P = 0.85). Calves-fed R30 tended to have greater ADG and final BW than calves-fed R60 (P ≤ 0.08). Gain efficiency (gain:feed ratio; G:F) was greater for calves-fed R30 than calves-fed R60 (P = 0.01). Dietary roughage level did not affect morbidity and mortality (P ≥ 0.11). Supplemental fat did not affect DMI (P = 0.6) but tended (P = 0.09) to increase ADG compared to -FAT diets. The G:F was greater for calves-fed +FAT than -FAT (P = 0.03). The +FAT diet tended (P = 0.10) to increase the number of retreatments against BRD compared to -FAT, although the total number of antimicrobial treatments required to treat sick calves (P = 0.78) and the mortality rate (P = 0.99) were not affected by supplemental fat. Feeding +FAT diet tended (P ≤ 0.09) to increase plasma concentration of cortisol and immunoglobulin-G compared to -FAT. In summary, feeding 30% roughage diets or adding 3.5% yellow grease (DM basis) as supplemental fat increased G:F during the feedlot receiving period.

The low intake of feed that beef calves exhibit during the first weeks after feedlot arrival results in inadequate nutrient intake, especially energy, which leads to low rates of gain and decreased immune function and likely increases the risks for respiratory diseases. Increasing the energy density of receiving diets (Mcal/kg of dry matter) could result in increased energy intake of newly received finishing cattle. In this experiment, we evaluated the effects of two roughage levels (wheat hay at 30% [R30] or 60% [R60]; dry matter [DM] basis) combined with two levels of supplemental fat (yellow grease at 0% [−FAT; no supplemental fat] or 3.5% [+FAT]; DM basis). Calves-fed R30 tended to have greater average daily gain and final body weight than calves-fed R60. Gain efficiency (gain:feed ratio; G:F) was greater for calves-fed R30 than calves-fed R60. Feeding +FAT tended to increase average daily gain compared to −FAT diet, and G:F was greater for calves-fed +FAT than −FAT. In summary, feeding 30% roughage diets or adding 3.5% of yellow grease increased G:F during the feedlot receiving period, with minimal impact on morbidity rate from respiratory disease.

Alterations in Skeletal Muscle mRNA Abundance in Response to Ethyl-Cellulose Rumen-Protected Methionine during the Periparturient Period in Dairy Cows.

This study aimed to evaluate the effect of feeding ethyl cellulose rumen-protected methionine (RPM) on skeletal muscle mRNA abundance during the periparturient period. Sixty multiparous Holstein cows were used in a block design and assigned to either a control or RPM diet. The RPM was supplied from −28 to 60 days in milk (DIM) at a rate of 0.09% (prepartum) or 0.10% (postpartum) of dry matter (DM), ensuring a Lys:Met in the metabolizable protein of ~2.8:1. Muscle biopsies were collected at −21, 1, and 21 DIM. Thirty-five target genes associated with nutrient metabolism and biochemical pathways were measured via RT-qPCR. The mRNA abundance of genes associated with amino acid (AA) transport (SLC7A8, SLC43A2), carnitine transport (SLC22A5), insulin signaling (IRS1), and antioxidant response (NFE2L2) had diet × time effect (p < 0.05) due to greater abundance in RPM versus CON cows, especially at 1 and 21 DIM. Members of the AA transport (SLC7A8, SLC25A29, SCL38A9), fatty acid ß-oxidation (ACADVL), vitamin transport (SLC5A6, SLC19A2), mTOR pathway (AKT1 and mTOR), antioxidant response (KEAP1, CUL3), CDP-Choline pathway and arginine metabolism had overall greater abundance (p < 0.05) in RPM versus CON cows. Overall, data indicate that RPM can alter nutrient metabolism in the skeletal muscle around parturition partly through alterations in mRNA abundance.

Effects of maternal dietary omega-3 polyunsaturated fatty acids and methionine during late gestation on fetal growth, DNA methylation, and mRNA relative expression of genes associated with the inflammatory response, lipid metabolism and DNA methylation in placenta and offspring's liver in sheep.

BACKGROUND: Omega-3 PUFA or methionine (Met) supply during gestation alters offspring physiology. However, the effect of both nutrients on fetal development has not been explored. Our objective was to determine the effects of supplementation of these two nutrients during late gestation on fetal growth, DNA methylation, and mRNA expression of genes associated with the inflammatory response, and DNA methylation. Ewes (n = 5/treatment) were fed from day 100 to 145 of gestation one of the following treatments: 1) basal diet (NS) without fatty acids (FS) or methionine (MS) supplementation; 2) FS (10 g/kg Ca salts, source omega-3 PUFA); 3) MS (1 g/kg rumen protected methionine); and 4) FS and MS (FS-MS). On day 145, ewes were euthanized, and data from dams and fetus was recorded. Placenta (cotyledon), fetal liver, and blood samples were collected. RESULTS: A treatments interaction on fetal liver weight, ewe body weight and body condition score (BCS) was observed; FS-MS were heavier (P <  0.01) than FS and MS, and FS-MS ewes had a better (P = 0.02) BCS than NS. Methionine increased (P = 0.03) ewe plasma glucose concentration. Fetal liver global DNA methylation increased (P <  0.01) in FS and MS. Dietary treatments modify the mRNA relative expression on some of the genes evaluated. In the fetal liver, FS increased (P = 0.04) the mRNA relative expression of arachidonate-5-lipoxygenase-activating-protein and tended to decrease (P = 0.06) methionine-adenosyltransferase-1A. Moreover, MS decreased (P = 0.04) DNA-methyltransferase-1 and tended to decrease (P = 0.08) free-fatty-acid-receptor-1 mRNA relative expression. Furthermore, FS-MS decreased mRNA relative expression of tumor-necrosis-factor-alpha (P = 0.05), peroxisome-proliferator-activated-receptor-delta (P = 0.03) and gamma (P = 0.04), tended to decrease (P ≤ 0.09) interleukin-6, fatty-acid-transport-protein-1, and delta-5-desaturase, and increased adenosylhomocysteinase (P = 0.04) mRNA relative expression. In cotyledon, FS tended to decrease fatty acid binding protein 4 (P = 0.09) mRNA relative expression. CONCLUSION: Omega-3 PUFA and Met supplementation improves dam's performance in late gestation, which was positively correlated with an increase in offspring's liver development. Moreover, FS-MS decreased mRNA relative expression of proinflammatory cytokines, and lipogenic genes, and increased the expression on an enzyme that has an important role in methylation.

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

BACKGROUND: Postruminal 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. OBJECTIVES: To 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. METHODS: Holstein 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. RESULTS: Cows 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 ß-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. CONCLUSIONS: Greater 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.

Maternal supply of methionine during late-pregnancy enhances rate of Holstein calf development in utero and postnatal growth to a greater extent than colostrum source.

BACKGROUND: Pregnancy and early life are critical periods of plasticity during which the fetus and neonate may be influenced by environmental factors such as nutrition. Maternal methionine (Met) supply in non-ruminants during pregnancy can affect offspring development and growth. Thus, the objective of this study was to investigate if increasing Met supply during late-pregnancy affects developmental parameters of the calf at birth and if either maternal Met or colostrum from Met-fed cows alters calf growth. Calves born to Holstein cows individually-fed a basal control [CON; 1.47 Mcal/kg dry matter (DM) and 15.3% crude protein] diet with no added Met or CON plus ethylcellulose rumen-protected Met (MET; Mepron® at 0.09% of diet DM; Evonik Nutrition & Care GmbH, Germany) during the last 28 ± 2 d of pregnancy were used. A total of 39 calves were in CON (n = 22 bulls, 17 heifers) and 42 in MET (n = 20 bulls, 22 heifers). At birth, calves were randomly allocated considering dam treatment and colostrum as follows: 1) calves from CON cows and colostrum from CON cows (n = 21); 2) calves from CON cows and colostrum from MET cows (n = 18); 3) calves from MET cows and colostrum from MET cows (n = 22); and 4) calves from MET cows and colostrum from CON cows (n = 20). All calves were housed, managed, and fed individually during the first 9 wk of life. RESULTS: Despite greater daily DM intake pre-partum in cows fed MET (15.7 vs. 14.4 ± 0.12 kg/d, P < 0.05), colostrum quality and quantity were not affected by maternal diet. At birth, MET calves had greater (P ≤ 0.05) body weight (BW, 44.1 vs. 42.1 ± 0.70 kg), hip height (HH, 81.3 vs. 79.6 ± 0.53 cm) and wither height (WH, 77.8 vs. 75.9 ± 0.47 cm). In contrast, concentrations of His, Lys, and Asn in plasma were lower (P ≤ 0.05) in MET calves. Regardless of colostrum source, the greater BW, HH, and WH in MET calves at birth persisted through 9 wk of age resulting in average responses of + 3.1 kg BW, + 1.9 cm HH, and + 1.8 cm WH compared with CON. Average daily gain during the 9 wk was (P < 0.05) 0.72 ± 0.02 kg/d in MET compared with 0.67 ± 0.02 kg/d in CON calves. Respiratory scores were normal and did not differ (P > 0.05) due to maternal Met supply or colostrum source. However, fecal scores tended to be lower (P ≤ 0.10) in MET calves regardless of colostrum source. CONCLUSIONS: Increasing the maternal supply of MET during late-pregnancy enhanced growth in utero as well as during the pre-weaning and early post-weaning periods. Although the ~ 1 kg/d greater DM intake during the last 2-3 wk prior to parturition could explain a portion of the 2 kg extra body mass of MET calves at birth, other mechanisms potentially encompassing nutrient assimilation efficiency likely played a role. Assessing the exact mechanisms sensitive to supply of Met or total amino acid supply during the latter stages of growth in utero merit further research.