Protein Supplementation during Mid-Gestation Alters the Amino Acid Patterns, Hepatic Metabolism, and Maternal Skeletal Muscle Turnover of Pregnant Zebu Beef Cows.

From 100 to 200 days of gestation, 52 cows carrying male (n = 30) or female (n = 22) fetuses were assigned to CON (basal diet-5.5% of CP, n = 26) or SUP (basal diet + protein supplement [40% CP, 3.5 g/kg BW]-12% of CP, n = 26) treatments. Glucose concentrations decreased at 200 (p ≤ 0.01; CON = 46.9 and SUP = 54.7 mg/dL) and 270 days (p ≤ 0.05; CON = 48.4 and SUP = 53.3 mg/dL) for CON compared to SUP. The same pattern occurred for insulin (p ≤ 0.01). At parturition, the NEFA concentration was greater (p = 0.01, 0.10 vs. 0.08 mmol/L) for CON than for SUP. Total AA increased in SUP (p ≤ 0.03) at mid- and late-gestation compared to CON. At 200 days, CON dams carrying females had less essential AA (p = 0.01) than cows carrying males. The SUP dams had greater expressions of protein synthesis markers, namely eIf4E and GSK3ß (p ≤ 0.04), at day 200 and of MuFR1 (protein degradation marker, p ≤ 0.04) at parturition. Supplemented cows had higher hepatic pyruvate carboxylase expressions (p = 0.02). Therefore, PS alleviates the restriction overload on maternal metabolism.

Effects of Maternal Protein Supplementation at Mid-Gestation of Cows on Intake, Digestibility, and Feeding Behavior of the Offspring.

This study aimed to assess the effects of maternal protein supplementation and offspring sex (OS) on the intake parameters of the offspring. Forty-three Tabapuã cows were randomly allocated in the following treatments: protein supplementation (PS) during days 100-200 of gestation (RES, 5.5% total crude protein (CP), n = 2, or CON, 10% total CP, n = 19) and OS (females, n = 20; males, n = 23). The offspring were evaluated during the cow-calf (0-210 days), backgrounding (255-320 days), growing 1 (321-381 days), and growing 2 (382-445 days) phases. The CON offspring tended to present higher dry matter intake (DMI) at weaning (p = 0.06). The CON males presented lower digestibility of major diet components in the growing 2 phase (p ≤ 0.02). The CON offspring spent 52% more time per day eating supplements at 100 days and 17% less time in idleness at 210 days. The CON males spent 15 min more per day ruminating than RES males in the feedlot phase (p = 0.01). We concluded that protein supplementation over gestation alters the offspring feed intake pattern as a whole, while protein restriction promotes compensatory responses on nutrient digestibility in males.

Impact of Maternal Feed Restriction at Different Stages of Gestation on the Proteomic Profile of the Newborn Skeletal Muscle.

We aimed to investigate the effects of the maternal plane of nutrition during gestation on the proteome profile of the skeletal muscle of the newborn. Pregnant goats were assigned to the following experimental treatments: restriction maintenance (RM) where pregnant dams were fed at 50% of their maintenance requirements from 8−84 days of gestation, and then feed of 100% of the maintenance requirements was supplied from 85­parturition (n = 6); maintenance restriction (MR) where pregnant dams were fed at 100% of their maintenance requirements from 8−84 days of gestation, and then experienced feed restriction of 50% of the maintenance requirements from 85­parturition (n = 8). At birth, newborns were euthanized and samples of the Longissimus dorsi muscle were collected and used to perform HPLC-MS/MS analysis. The network analyses were performed to identify the biological processes and KEGG pathways of the proteins identified as differentially abundant protein and were deemed significant when the adjusted p-value (FDR) < 0.05. Our results suggest that treatment RM affects the energy metabolism of newborns' skeletal muscle by changing the energy-investment phase of glycolysis, in addition to utilizing glycogen as a carbon source. Moreover, the RM plane of nutrition may contribute to fatty acid oxidation and increases in the cytosolic α-KG and mitochondrial NADH levels in the skeletal muscle of the newborn. On the other hand, treatment MR likely affects the energy-generation phase of glycolysis, contributing to the accumulation of mitochondrial α-KG and the biosynthesis of glutamine.

L-Arginine Supplementation for Nulliparous Sows during the Last Third of Gestation.

We evaluated the effects of L-arginine supplementation during the last third of gestation on molecular mechanisms related to skeletal muscle development of piglets and litter traits at birth. Twenty-three nulliparous sows averaging 205.37 ± 11.50 kg of body weight were randomly assigned to the following experimental treatments: control (CON), where pregnant sows were fed diets to meet their nutritional requirements; arginine (ARG), where sows where fed CON + 1.0% L-arginine. Skeletal muscle from piglets born from sows from ARG group had greater mRNA expression of MYOD (p = 0.043) and MYOG (p ≤ 0.01), and tended to present greater mRNA expression (p = 0.06) of IGF-2 gene compared to those born from CON sows. However, there were no differences (p > 0.05) in the histomorphometric variables of fetuses' skeletal muscle. The total weight of born piglets, total weight of born alive piglets, piglet weight at birth, coefficient of variation of birth weight, and the incidence of intrauterine growth restriction (IUGR) piglets did not differ between groups. No stillborn piglets (p < 0.01) were verified in the ARG sows compared to CON group. The blood levels of estradiol (p = 0.035) and urea (p = 0.03) were higher in ARG sows compared to those from the CON group. In summary, our data show that arginine supplementation of nulliparous sows at late gestation enhance mRNA expression of key myogenic regulatory factors, which likely contribute to improve animal growth rates in later stages of development.

Proteomic Analysis of Liver from Finishing Beef Cattle Supplemented with a Rumen-Protected B-Vitamin Blend and Hydroxy Trace Minerals.

Vitamin B and trace minerals are crucial molecular signals involved in many biological pathways; however, their bioavailability is compromised in high-producing ruminant animals. So far, studies have mainly focused on the effects of these micronutrients on animal performance, but their use in a rumen-protected form and their impact on liver metabolism in finishing beef cattle is poorly known. We used a shotgun proteomic approach combined with biological network analyses to assess the effects of a rumen-protected B-vitamin blend, as well as those of hydroxy trace minerals, on the hepatic proteome. A total of 20 non-castrated Nellore males with 353 ± 43 kg of initial body weight were randomly assigned to one of the following treatments: CTRL-inorganic trace minerals without supplementation of a protected vitamin B blend, or SUP-supplementation of hydroxy trace minerals and a protected vitamin B blend. All animals were fed the same amount of the experimental diet for 106 days, and liver biopsies were performed at the end of the experimental period. Supplemented animals showed 37 up-regulated proteins (p < 0.10), and the enrichment analysis revealed that these proteins were involved in protein folding (p = 0.04), mitochondrial respiratory chain complex I (p = 0.01) and IV (p = 0.01), chaperonin-containing T-complex 2 (p = 0.01), glutathione metabolism (p < 0.01), and other aspects linked to oxidative-stress responses. These results indicate that rumen-protected vitamin B and hydroxy trace mineral supplementation during the finishing phase alters the abundance of proteins associated with the electron transport chain and other oxidation-reduction pathways, boosting the production of reactive oxygen species, which appear to modulate proteins linked to oxidative-damage responses to maintain cellular homeostasis.

Skeletal Muscle Development in Postnatal Beef Cattle Resulting from Maternal Protein Restriction during Mid-Gestation.

We aimed to investigate the effects of maternal protein restriction during mid-gestation on the skeletal muscle composition of the offspring. In the restriction treatment (RES, n = 9), cows were fed a basal diet, while in the control (CON, n = 9) group cows received the same RES diet plus the protein supplement during mid-gestation (100-200d). Samples of Longissimus dorsi muscle were collected from the offspring at 30d and 450d postnatal. Muscle fiber number was found to be decreased as a result of maternal protein restriction and persisted throughout the offspring's life (p < 0.01). The collagen content was enhanced (p < 0.05) due to maternal protein restriction at 30d. MHC2X mRNA expression tended to be higher (p = 0.08) in RES 30d offspring, however, no difference (p > 0.05) was found among treatments at 450d. Taken together, our results suggest that maternal protein restriction during mid-gestation has major and persistent effects by reducing muscle fiber formation and may slightly increase collagen accumulation in the skeletal muscle of the offspring. Although maternal protein restriction may alter the muscle fiber metabolism by favoring the establishment of a predominant glycolytic metabolism, the postnatal environment may be a determinant factor that establishes the different proportion of muscle fiber types.

Intramuscular collagen characteristics and expression of related genes in skeletal muscle of cull cows receiving a high-energy diet.

We aimed to investigate differences in the synthesis and metabolism of intramuscular collagen in the Longissimus thoracis (LT) muscle between heifers and cull-cows fed high-energy diet. Ten cull-cows, (74.9 ± 3.2 months age, weighing 536 ± 14.55 kg) and ten heifers (18.4 ± 3.2 months age, weighting 310.5 ± 14.5 kg) were fed with high-energy diets for 150 days. The total collagen content did not differ between treatments. Greater collagen solubility was observed in heifers than cull-cows, although no differences in lysyl oxidase activity were observed between treatments. No differences were observed for mRNA expression of CO1A1, MMP2, MMP9 and TIMP2 between treatments. However, cull-cows presented greater mRNA expression of COL3A1, TIMP1 and TIMP3 than heifers. Our data give no indication that feeding a high-energy diet to cull-cows decreases the concentration of intramuscular collagen in the LT muscle or increases its solubility in respect to the collagen solubility in LT muscles from heifers on the same diet.

Effects of energy-protein supplementation frequency on performance of primiparous grazing beef cows during pre and postpartum.

OBJECTIVE: Twenty-four pregnant Nellore primiparous grazing cows were used to evaluate the effects of energy-protein supplementation and supplementation frequency during pre (105 d before calving) and postpartum (105 d after calving) on performance and metabolic characteristics. METHODS: Experimental treatments consisted of a control (no supplementation), daily supplementation (1.5 kg/d of concentrate/animal) and infrequent supplementation (4.5 kg of concentrate/animal every three days). During the pre and postpartum periods, concentrations of blood metabolites and animal performance were evaluated. Ureagenesis and energy metabolism markers were evaluated at prepartum period. RESULTS: Supplementation frequency did not alter (p>0.10) body weight (BW), average daily gain (ADG), and carcass traits during pre and postpartum. The BW (p = 0.079), adjusted BW at day of parturition (p = 0.078), and ADG (p = 0.074) were greater for supplemented cows during the prepartum. The body condition score (BCS; p = 0.251), and carcass traits (p>0.10) were not affected by supplementation during prepartum. On postpartum, supplementation did not affect animal performance and carcass traits (p>0.10). The dry mater intake was not affected (p>0.10) by supplementation and supplementation frequency throughout the experimental period. Daily supplemented animals had greater (p<0.001) glucose levels than animals supplemented every three days. Supplementation and supplementation frequency did not alter (p>0.10) the levels of blood metabolites, neither the abundance of ureagenesis nor energy metabolism markers. CONCLUSION: In summary, our data show that the reduction of supplementation frequency does not cause negative impacts on performance and metabolic characteristics of primiparous grazing cows during the prepartum.

Guanidinoacetic acid supplementation on growth performance and molecular mechanisms of lean mass gain in nursery pigs / Suplementação de ácido guanidinoacético no desempenho e mecanismo moleculares de ganho de massa magra em leitões

ABSTRACT: The objective with the present study was to evaluate the effect of guanidinoacetic acid (GAA) on the growth performance of nursery piglets as well as a possible molecular mechanism of action on lean mass gain. Seventy-two pigs, weaned at 21 d, weighing 6.80 ± 1.2 kg were distributed in a completely randomized design into one of three dietary treatments (control, control + 1.2 g/kg GAA or control + 2.4 g/kg GAA) and 8 replicates per treatment. The control diet was an animal protein-free diet based on corn and soybean meal. Body weight, average daily weight gain, average daily feed intake and feed efficiency were evaluated at 35, 49, and 56 days. At the end of the experiment, one animal per pen was slaughtered and samples of the vastus lateralis muscle were collected for RT-qPCR and protein abundance analysis. Overall (from 21 to 56 d), GAA supplementation improved feed efficiency (P < 0.03). Skeletal muscle of pigs fed with GAA diet had greater mRNA expression of Akt (P < 0.04) and RPS6KB2 (P<0.01). In conclusion, supplementation with 2.4 g/kg GAA to nursery piglets improves feed efficiency and activates molecular mechanisms important to lean mass gain.

RESUMO: O objetivo do presente estudo foi avaliar o efeito do ácido guanidinoacético (GAA) no desempenho de leitões, bem como um possível mecanismo de ação molecular no ganho de massa magra. Setenta e dois leitões, desmamados aos 21 dias, pesando 6,80 ± 1,2 kg, foram distribuídos em um delineamento inteiramente casualizado com três tratamentos dietéticos (controle, controle + 1,2 g / kg ou controle + 2,4 g / kg GAA) e 8 repetições por tratamento. A dieta controle não continha proteína animal e foi formulada a base de milho e farelo de soja. O desempenho dos animais foi avaliado aos 35, 49 e 56 dias. Ao final do experimento, um animal por unidade experimental foi abatido e amostras do músculo Vastus lateralis foram coletadas para análise de RT-qPCR e abundância de proteínas. A suplementação com GAA melhorou a eficiência alimentar (P<0,03) aos 56 dias. O músculo dos leitões suplementados apresentou maior expressão de mRNA de Akt (P<0,04) e RPS6KB2 (P <0,01). Em conclusão, a suplementação de 2,4 g / kg de GAA em leitões (21 a 56 d) melhora a eficiência alimentar e ativa mecanismos moleculares importantes para o ganho de massa magra.

The Bacterial and Fungal Microbiota of Nelore Steers Is Dynamic Across the Gastrointestinal Tract and Its Fecal-Associated Microbiota Is Correlated to Feed Efficiency.

The ruminant gastrointestinal tract (GIT) microbiome plays a major role in the health, physiology and production traits of the host. In this work, we characterized the bacterial and fungal microbiota of the rumen, small intestine (SI), cecum and feces of 27 Nelore steers using next-generation sequencing and evaluated biochemical parameters within the GIT segments. We found that only the bacterial microbiota clustered according to each GIT segment. Bacterial diversity and richness as well as volatile fatty acid concentration was lowest in the SI. Taxonomic grouping of bacterial operational taxonomic units (OTUs) revealed that Lachnospiraceae (24.61 ± SD 6.58%) and Ruminococcaceae (20.87 ± SD 4.22%) were the two most abundant taxa across the GIT. For the fungi, the family Neocallismastigaceae dominated in all GIT segments, with the genus Orpinomyces being the most abundant. Twenty-eight bacterial and six fungal OTUs were shared across all GIT segments in at least 50% of the steers. We also evaluated if the fecal-associated microbiota of steers showing negative and positive residual feed intake (n-RFI and p-RFI, respectively) was associated with their feed efficiency phenotype. Diversity indices for both bacterial and fungal fecal microbiota did not vary between the two feed efficiency groups. Differences in the fecal bacterial composition between high and low feed efficiency steers were primarily assigned to OTUs belonging to the families Lachnospiraceae and Ruminococcaceae and to the genus Prevotella. The fungal OTUs shared across the GIT did not vary between feed efficiency groups, but 7 and 3 OTUs were found only in steers with positive and negative RFI, respectively. These results provide further insights into the composition of the Nelore GIT microbiota, which could have implications for improving animal health and productivity. Our findings also reveal differences in fecal-associated bacterial OTUs between steers from different feed efficiency groups, suggesting that fecal sampling may represent a non-invasive strategy to link the bovine microbiota with productivity phenotypes.

Effects of grain processing methods on the expression of genes involved in volatile fatty acid transport and pH regulation, and keratinization in rumen epithelium of beef cattle.

Two experiments were carried out to evaluate the effects of corn and sorghum with different processing methods on the expression of genes involved in volatile fatty acids transport and pH regulation, and ruminal keratinization in rumen epithelium of finishing bulls. For Exp. 1, five rumen cannulated Nellore bulls were used in a 5x5 Latin square arrangement, with 14 d for adaptation and 9 d for sample collection. Treatments were: dry ground corn, dry ground sorghum, reconstituted corn, reconstituted sorghum, and control (forage-based diet). Samples of rumen epithelium from ventral sac were excised, rinsed, snap-frozen and stored at -80°C until total RNA isolation and quantitative real-time PCR analysis. In the Exp. 2, 24 Nellore bulls were assigned to a completely randomized design lasting 168 d. Experimental treatments were similar to those at Exp. 1, but without the control treatment. After the experimental period, bulls were slaughtered and rumen epithelium samples were rapidly excised for further histological analysis. Rumen epithelial tissue from animals fed reconstituted corn had lower expression of downregulated-in-adenoma (P = 0.03) and Na+/H+ exchanger 2 (trend; P = 0.09). The expression of Na+/ H+ exchanger 1 (P = 0.10) and putative anion transporter (P = 0.06) tended to be lower in rumen epithelium of bulls fed reconstituted grains. Ruminal concentration of valerate was greater for animals fed reconstituted grain (P = 0.01). Likewise, animals fed reconstituted corn tended to have greater butyrate ruminal concentration (P = 0.08). Keratinized layer thickness did not differ among treatments (P > 0.10). Therefore, reconstituted grains (especially corn) decrease the mRNA expression of genes involved in volatile fatty acids transport and pH control in the rumen epithelium.

Effect of feeding strategies on weaning weight and milk production of Holstein × Zebu calves in dual purpose milk production systems.

The objective of this study was to evaluate the effect of five feeding strategies on calf weaning weight, and cow milk production and composition in Brazilian Holstein × Zebu cows. A total of 60 cows and their calves were allocated to each of five treatments. Cows in treatments 1, 2 and 3 were milked for 270 days and cows in treatments 4 and 5 were milked for 180 days. Calves in treatment 1 (CON) were not supplemented with concentrate whereas calves from treatment 2 (CLPN) received 1 kg of concentrate daily from 90 to 270 days of age and calves from treatment 3 received 1 kg of concentrated from 180 to 270 days of age. Calves in treatment 4 (CCPS) were supplemented with 1 kg of concentrate from 90 to 180 days of age and calves in treatment 5 (CLPS) were supplemented with 1 kg of concentrate from 90 to 270 days of age. Calves from the CLPS treatment had greater milk and protein intakes (P < 0.05) and greater growth rate than calves from the other treatments. Our results indicate that the traditional system of feeding calves with no concentrate results in a weight gain of 600 g/day. The CLPS treatment produced calves with the highest live weight and growth rate. The nutritional strategy with restricted supply of milk for the calves with concomitantly short-term concentrate supplementation does not improve performance of calves but did increase feed costs.

Strategies of supplementation of female suckling calves and nutrition parameters of beef cows on tropical pasture.

The performance of female calves in creep feeding under different strategies of supplementation and milk production, intake, and digestibility of grazing Nellore and crossbred cows (Nellore × Holstein) during the dry-rainy transition season were assessed. Forty-four female beef suckling calves, with initial age between 90 and 150 days and average initial body weight of 117.7 ± 4.3 kg, and their respective dams (24 Nellore and 20 crossbred) with average initial body weight of 417.5 ± 8.3 kg, were used. The experimental treatments consisted of: control group--mineral mixture only; strategy 1--supplementation from 112 days prior to weaning (0.375 kg/animal/day); strategy 2--supplementation from 112 days prior to weaning, in increasing amounts of 0.15, 0.30, 0.45, and 0.60 kg/animal/day through the four experimental periods, respectively; and strategy 3--supplementation from 56 days prior to weaning (0.750 kg/animal/day). Calves from strategy 1 had greater (P < 0.05) average daily gain (0.672 kg/day) than control animals (0.582 kg/day) and greater (P < 0.05) efficiency of supplement use than the other groups. Crossbred cows produced more milk than Nellore cows (P < 0.05). Crossbred cows presented greater (P < 0.05) dry matter intake (DMI) than Nellore cows. However, no differences were found (P > 0.05) for nutrient digestibility among genetic types. It can be concluded that strategies of supplementation that present an equitable distribution of supplement provides greater weight gain in suckling female beef calves. Crossbred cows produce more milk and present greater DMI than Nellore cows. There are no differences in the nutrient digestibility between Nellore and Nellore × Holstein crossbred cows.

Intake, digestibility, performance, and carcass traits of beef cattle of different gender.

The performance, intake, feed efficiency, and carcass traits of beef cattle from different gender profile were assessed. Fifteen animals (five steers, five spayed heifers, and five intact heifers) with ±250 kg of initial body weight were randomly assigned in individual pens and fed the same diet for 106 days. At the end of the trial, all the animals were slaughtered and the pH, temperature, and weight of the carcass were recorded. The right side of each carcass was then separated into chuck, shoulder, flank sirloin, and round for evaluation of commercial cuts yield. The left carcass sides were ribbed between the 12th and 13th ribs where the rib eye area and fat thickness measurements were taken. The 9th-11th rib section was removed from the left half carcass and then dissected into muscle, fat, and bones in order to estimate carcass composition. Gender had no effect (P > 0.05) on performance, intake, digestibility of dry matter and all the nutrients evaluated, feed efficiency, and carcass characteristics. It can be concluded that steers and heifers (spayed or not) have the same potential to produce beef. From a productive and welfare standpoint, there is no reason to spay heifers.

Performance and meat quality traits of beef heifers fed with two levels of concentrate and ruminally undegradable protein.

The effects of two levels of concentrate and ruminally undegradable protein (RUP) on performance, intake, digestibility, carcass characteristics, meat quality traits, and commercial cuts yield were assessed. Twenty crossbred heifers (240 kg average body weight) were used. At the beginning of the trial, four animals were slaughtered as reference group and the 16 remaining animals were randomly assigned to four treatments, in a 2 × 2 factorial design: two levels of concentrate (40% and 80%, dry matter (DM) basis) and two levels of RUP (48.79% and 27.19% of CP). At the end of the trial, all the animals were slaughtered. There was no interaction (P > 0.05) between concentrate and RUP levels. Dry matter intake and nutrients digestibility was not affected (P > 0.05) by RUP level. Heifers fed the highest RUP level had greater (P < 0.05) average daily gain (ADG). Concentrate level did not affect (P > 0.05) DMI and ADG. Heifers fed diets with 80% concentrate had greater intake of TDN and EE, and lower intake of NDF (P < 0.05). The digestibilities of all nutrients, except the NDF, were greater (P < 0.05) for the 80% concentrate diets. The RUP levels did not affect (P > 0.05) the carcass characteristics and carcass gain composition. Heifers fed 80% concentrate diets had larger (P < 0.05) longissimus muscle area and lower gain of bones (P < 0.05). Concentrate level did not affect (P > 0.05) the composition of carcass gain. There was no effect (P > 0.05) of RUP and concentrate levels on meat quality traits and commercial cut yields.