Protein requirements for pregnant dairy cows.

We aimed to estimate the protein requirements of pregnant Holstein × Gyr cows. A total of 61 Holstein × Gyr (HG) cows were used, with an average initial body weight (BW) of 480 ± 10.1 kg and age 5 ± 0.5 yr. Cows were divided into 3 groups: pregnant (n = 43), nonpregnant (n = 12), and baseline (n = 6). Baseline animals were slaughtered before starting the experiment to estimate the initial body composition of the remaining animals. Pregnant and nonpregnant cows received 2 diets: maintenance and ad libitum. Pregnant cows were slaughtered at 139, 199, 241, and 268 gestation days (GD). First, we used data only from nonpregnant cows to determine the requirements for maintenance and growth in adult cows. The requirements of metabolizable protein for maintenance (MPm; grams of empty metabolic BW [EBW0.75] per day) were estimated using a linear regression between the metabolizable protein intake (MPI, g/d) and average daily gain (g/d), and the MPm was defined as the intercept divided by the average metabolic BW. Net protein requirements for gain (NPg; g/d) were estimated by the first derivative of the allometric equation between final crude protein in the body (kg) and the final empty BW (EBW; kg). The efficiency of use of metabolizable protein for gain (k) was calculated from the regression between the retained protein (g EBW0.75/d) and the MPI (g EBW0.75/d), and k was the slope of this regression. The MPI was estimated by combining microbial protein synthesis (purine derivatives) with the digestible rumen undegradable protein [(total protein intake - rumen-degradable protein) × intestinal digestibility]. Second, an exponential model was used to fit the protein accumulation in the gestational components in the function of GD. The first derivative of that model was considered the net requirement for gestation (NPgest). The efficiency of protein utilization for gestation (kgest) was calculated by the iterative method using the equation Δ = MPI - (MPm + NPg/kg + NPgest/kgest), where kg is efficiency of protein utilization for gain. The iteration was performed aiming at a zero deviation between observed MPI and metabolizable protein (MP) estimated by the requirements determined herein. We obtained a value of 3.88 g EBW0.75/d for MPm. The estimation of NPg can be calculated according to the following equation: NPg = 0.716 × (EBWopen-0.308) × EBGc, where EBWopen is the EBW (kg) for nonpregnant animals and EBGc is the empty body gain (kg/d) corrected for the gestational component. The k was determined as 0.347. The NPgest requirements were determined as NPgest (g/d) = 0.0008722 × exp(0.01784 × GD) × (calf weight/35). The kgest was 0.625. It is important to highlight that different methods of MP estimates should not be mixed and that the proposed method requires the estimation of microbial protein (estimated via urinary estimates), which might limit practical application. In conclusion, new studies should be conducted to validate our results and the methodology adopted to determine protein requirements for pregnancy in dairy cows. Due to the pattern of protein accumulation in the gestational components, we suggest an exponential model to describe protein requirements for pregnancy for dairy cows.

Intake, performance, and feeding behavior of Holstein and Holstein × Gyr heifers grazing intensively managed tropical grasses during the rainy season.

Holstein × Gyr and Holstein are the primary dairy breeds used in tropical systems, but when rearing under pasture, feed intake, behavior, and performance might differ between them. This study aimed to evaluate the voluntary intake, nutrient digestibility, performance, and ingestive behavior of Holstein and Holstein × Gyr (½ Holstein × ½ Gyr) heifers managed in a rotational system of Guinea grass (Panicum maximum Jacq. cv. Mombaça). The experiment was conducted during the summer season throughout four periods of 21 d. Two 8-heifers (four Holstein and four Holstein × Gyr) groups, averaging 258.6 ±â€¯24.79 kg and 157.1 ±â€¯24.99 kg BW, were used. Each group grazed a separate set of 16 paddocks, and all heifers received a concentrate supplement daily. Heifers were weighed at the beginning and end of the experiment. Fecal, forage and concentrate samples were evaluated for their DM, CP, crude fat, ash, NDF, and indigestible NDF. Feeding behavior was evaluated through 24 h of live observation for 48 h of each experimental period. Grazing, ruminating, resting, and intake of concentrate times were recorded, and rumination criteria, bout criteria, mealtime, meal frequency, and meal duration were calculated. There was no difference in total dry matter intake (DMI), but forage DMI of Holstein × Gyr was 11.70 % greater than the Holstein heifers. The Holstein × Gyr heifers had greater NDF intake and feed efficiency tended to show greater CP and NDF digestibilities, consequently, they had greater average daily gain (ADG). Holstein grazed less than Holstein × Gyr heifers in the afternoon. Ruminating time was 18.43 % lower for Holstein than Holstein × Gyr heifers, and rumination criteria (i.e. longest non-feeding interval within a rumination event) were greater for Holstein heifers. Holstein heifers presented more prolonged rumination bouts and resting time than Holstein × Gyr heifers. Holstein × Gyr can ingest and ruminate greater amounts of fibrous material, and Holstein heifers needed to spend more time ruminating the cud. Overall, even though the behavior was not markedly different between breeds, rearing young Holstein heifers in tropical pasture conditions is less suitable than Holstein-Gyr because of their lower ADG. Therefore, this management condition seems appropriate for Holstein × Gyr but inappropriate for Holstein dairy heifers.

Influence of Parity on the Colostrum Bacterial Community Composition in Holstein Cows.

The colostrum bacteriome of primiparous (P) and multiparous (M) Holstein cows was analysed by 16S rRNA sequencing. The species richness (Chao 1), diversity (Shannon and Simpson), and beta diversity did not differ between cow groups. The phyla Firmicutes, Tenericutes, Kiritimatiellaeota, and Fibrobacteres were more abundant in M cows, while Proteobacteria, Actinobacteria, Cloacimonetes, and Fusobacteria were more abundant in P cows. At the genus level, no significant differences were observed between groups (P < 0.05), and Prevotella_1 was the most abundant taxon. P and M cows shared 1030 taxonomic operational units (OTUs), with Acinetobacter showing greater abundance. In conclusion, parity did not affect the colostrum bacteriome of P and M cows and a healthy mammary gland could represent a reservoir of Acinetobacter in dairy farms. Supplementary Information: The online version contains supplementary material available at 10.1007/s12088-022-01016-x.

Energy and protein requirements of Holstein × Gyr crossbred heifers.

Nutrient requirements in cattle are dependent on physiological stage, breed and environmental conditions. In Holstein × Gyr crossbred dairy heifers, the lack of data remains a limiting factor for estimating energy and protein requirements. Thus, we aimed to estimate the energy and protein requirements of Holstein × Gyr crossbred heifers raised under tropical conditions. Twenty-two crossbred (½ Holstein × ½ Gyr) heifers with an average initial BW of 102.2 ± 3.4 kg and 3 to 4 months of age were used. To estimate requirements, the comparative slaughter technique was used: four animals were assigned to the reference group, slaughtered at the beginning of the experiment to estimate the initial empty BW (EBW) and composition of the animals that remained in the experiment. The remaining animals were randomized into three treatments based on targeted rates of BW gain: high (1.0 kg/day), low (0.5 kg/day) and close to maintenance (0.1 kg/day). At the end of the experiment, all animals were slaughtered to determine EBW, empty body gain (EBG) and body energy and protein contents. The linear regression parameters were estimated using PROC MIXED of SAS (version 9.4). Estimates of the parameters of non-linear regressions were adjusted through PROC NLIN of SAS using the Gauss-Newton method for parameter fit. The net requirements of energy for maintenance (NEm) and metabolizable energy for maintenance (MEm) were 0.303 and 0.469 MJ/EBW0.75 per day, respectively. The efficiency of use of MEm was 64.5%. The estimated equation to predict the net energy requirement for gain (NEg) was: NEg (MJ/day) = 0.299 × EBW0.75 × EBG0.601. The efficiency of use of ME for gain (kg) was 30.7%. The requirement of metabolizable protein for maintenance was 3.52 g/EBW0.75 per day. The equation to predict net protein requirement for gain (NPg) was: NPg (g/day) = 243.65 × EBW-0.091 × EBG. The efficiency of use of metabolizable protein for gain (k) was 50.8%. We observed noteworthy differences when comparing to ME and protein requirements of Holstein × Gyr crossbred heifers with other systems. In addition, we also observed differences in estimates for NEm, NEg, NPg, kg and k. Therefore, we propose that the equations generated in the present study should be used to estimate energy and protein requirements for Holstein × Gyr crossbred dairy heifers raised in tropical conditions in the post-weaning phase up to 185 kg of BW.

Determination of macromineral requirements for preweaned dairy calves in tropical conditions.

International committees that have published nutrient requirements for dairy cattle have used data from mineral studies conducted in the 1920s to 1970s, and no study has reported data from animals less than 100 kg; therefore, there is a need to update mineral requirements for preweaned dairy calves. Thus, a meta-analysis was performed to estimate the mineral requirements of Ca, P, K, Mg, and Na for Holstein and Holstein × Gyr crossbred preweaned dairy calves using data from 5 studies developed at the Universidade Federal de Viçosa (Viçosa, MG, Brazil). A total of 210 calves were separated into 2 breeds: purebred Holstein calves (animals with a Holstein pedigree higher than 87.5%) and Holstein × Gyr crossbred calves (animals with a Holstein pedigree lower than 87.5%). The comparative slaughter technique was used to estimate animal body composition and empty body weight (EBW). Mineral requirements for maintenance were estimated by the regression between retained mineral and mineral intake, whereas mineral requirements for gain were obtained from the first derivative of the mineral content in the animal's body. In addition, breed effect was tested on the intercept and slope of the models. The effect of breed was not observed for all analyzed variables. Thus, net requirements for maintenance were 12.73, 11.81, 20.28, 3.50, and 6.37 mg/kg of EBW per day for Ca, P, K, Mg, and Na, respectively. Retention coefficients were 73.18, 65.20, 13.16, 29.55, and 24.28% for Ca, P, K, Mg, and Na, respectively. The following equations were determined to estimate net requirements for gain (NRG, g/d): NRG for Ca = 14.402 × EBW-0.139 × empty body gain (EBG); NRG for P = 5.849 × EBW-0.027 × EBG; NRG for K = 1.140 × EBW-0.048 × EBG; NRG for Mg = 0.603 × EBW-0.036 × EBG; and NRG for Na = 1.508 × EBW-0.045 × EBG. Due to the high variation between the data found in this study and in the available literature, we suggest that further studies should be conducted to evaluate the estimates of this study.

Foetal development of skeletal muscle in bovines as a function of maternal nutrition, foetal sex and gestational age.

To determine the effects of maternal nutrition on modifications of foetal development of the skeletal muscle and possible increase in the potential of skeletal muscle growth in cattle, gestating cows were either fed 190% NRC recommendations (overnourished; ON) or 100% NRC recommendation (control; CO). Interaction between maternal nutrition (MN) and the foetal sex (FS) was also investigated. Foetuses were necropsied at four different time points throughout gestation (139, 199, 241 and 268 days of gestation) to assess the mRNA expression of myogenic, adipogenic and fibrogenic markers in skeletal muscle. Phenotypic indicators of the development of skeletal muscle fibres, intramuscular lipogenesis and collagen development were also evaluated. Modifications in mRNA expression of skeletal muscle of foetuses were observed in function of MN and FS despite the lack of effect of MN and FS on foetal weight at necropsy. Maternal ON increased the mRNA expression of the myogenic marker Cadherin-associated protein, beta 1 (CTNNB1) and adipogenic markers Peroxissome proliferator-activated receptor gamma (PPARG) and Zinc finger protein 423 (ZNF423) at midgestation. However, no differences on foetal skeletal muscle development were observed between treatments at late gestation indicating that a compensatory development may have occurred on CO foetuses making the effect of MN on skeletal muscle development not significant at late gestation. Moreover, our data have shown an evidence of sexual dimorphism during foetal stage with a greater skeletal muscle development in male than in female foetuses. In conclusion, providing a higher nutritional level to pregnant cows changes the trajectory of the development of skeletal muscle during midgestation, but apparently does not change the potential of post-natal growth of muscle mass of the offspring, as no differences in skeletal muscle development were observed in late gestation.

Energy and protein requirements for growth of Holstein × Gyr heifers.

There is little information regarding the nutritional requirements for dairy heifers, leading the majority of nutrient requirement systems to consider dairy heifers to be similar to beef heifers. Therefore, we evaluated the muscle protein metabolism and physical and chemical body composition of growing Holstein × Gyr heifers and estimated the energy and protein requirements. We performed a comparative slaughter experiment with 20 Holstein × Gyr heifers at an initial body weight of 218 ± 36.5 kg and an average age of 12 ± 1.0 months. Four heifers were designated as the reference group, and the 16 remaining heifers were fed ad libitum. The 16 heifers were distributed using a completely randomized design in a 2 × 2 factorial arrangement with two roughages (corn silage or sugarcane) and two concentrate levels (30 or 50%) for 112 days. Greater (p < 0.05) values for fractional rates of muscle protein synthesis, degradation and accretion were observed for heifers that were fed 50% concentrate. The following equations were obtained to estimate the net energy for gain (NEg ) and net protein for gain (NPg ): NEg (Mcal/day) = 0.0685 × EBW0.75  × EBWG1.095 and NPg (g/day) = 203.8 × EBWG - 14.80 × RE, respectively, in which EBW is the empty body weight, EBWG is the empty body weight gain and RE is the retained energy. We concluded that increased rates of protein turnover are achieved when a greater quality diet is provided. In the future, these results can be used to calculate the nutritional requirements for growth of Holstein × Gyr heifers after equation validation rather than using the recommendations provided by other systems, which use values developed from beef heifers, to determine the nutritional requirements of dairy cattle.

Does microbial nitrogen contamination affect the estimation of crude protein degradability of concentrate feeds?

The effects of microbial contamination (MC) on CP degradability of concentrate feeds are still controversial. Therefore, the aim of this study was to use N to estimate the impact of MC on estimations of CP fractions (the soluble fraction of CP [a], the insoluble but potentially degradable fraction of CP [b], and the rate of digestion of fraction b [kd]) of concentrate feeds. Twelve types of feed were evaluated: 6 energy concentrates-wheat bran ( L.), rice meal ( L.), ground corn ( L.), ground sorghum ( Pers.), ground corn cob ( L.), and soybean hulls [ (L.) Merr.]-and 6 protein concentrates-cottonseed meal ( L.), soybean meal [ (L.) Merr.], ground bean ( L.), peanut meal ( L.), sunflower meal ( L.), and corn gluten meal ( L.). The feeds were divided into 4 groups and were incubated in the rumen of 4 crossbred bulls. The samples were incubated for 0, 2, 4, 8, 16, 24, 48, and 72 h. To estimate the MC of the incubated residues, the ruminal bacteria were labeled with N via continuous intraruminal infusion of (NH)SO. There was no difference ( = 0.738) between corrected and uncorrected parameters a, b, and kd for all feeds that were evaluated. All of the feed tests followed an exponential model of degradation, and the model fitted well to the data, except for corn gluten meal, probably because the maximum incubation time that was used (72 h) was not long enough to allow for an accurate estimation of the degradation profile. Therefore, correction of ruminal protein degradation to MC is irrelevant with regards to the concentrates that were studied.

Intestinal development of bovine foetuses during gestation is affected by foetal sex and maternal nutrition.

We aimed to evaluate the effects of maternal nutrition (MN) and foetal sex on the intestinal development of bovine foetuses throughout different days of gestation (DG). Forty-four multiparous, dry Holstein × Gyr cows with average initial body weight of 480 ± 10 kg were fed the same diet of either restricted feeding at 1.15% of body weight (CO, n = 24) or fed ad libitum (overnourished, ON, n = 20). Six cows from CO group and five cows from ON group were slaughtered at 139, 199, 241 and 268 DG, and foetuses were necropsied to evaluate the intestinal development. The mass, length and density of foetal intestines were not affected by MN (p ≥ 0.260). An interaction between MN and DG was observed for the villi length of jejunum (p = 0.006) and ileum (p < 0.001). Villi length of jejunum and ileum was higher (p < 0.10) in foetuses from ON-fed cows than in foetuses from CO-fed cows at 139 DG. However, at 199 DG, the villi length of jejunum and ileum of foetuses from CO-fed cows was higher than in foetuses from ON-fed cows. Despite these differences, MN did not affect the villi length of jejunum and ileum at 268 DG (p > 0.10). Female foetuses had greater small intestine mass (p = 0.093), large intestine mass (p = 0.022), small intestine mass in proportion to body mass (p = 0.017) and large intestine mass in proportion to body mass (p < 0.001) than male foetuses. Female foetuses had also longer small intestine (p = 0.077) and greater small intestine density (p = 0.021) and villi length of jejunum (p = 0.001) and ileum (p = 0.010) than males. We conclude that MN affects the pathway for the development of foetal villi length throughout the gestation in bovine foetuses without changing the final villi length. Female foetuses had higher intestinal mass, density and villi length than males during the foetal phase in bovines.

Effect of maternal nutrition and days of gestation on pituitary gland and gonadal gene expression in cattle.

This study investigated effects of maternal overnutrition on gonadal development and pituitary-gonadal gene expression in cattle fetuses at mid- and late-gestation. Twenty-seven multiparous dry cows were fed either high (ad libitum, H) or moderate (M) intake of the same diet. Twelve cows from H (n=6) and M (n=6) intake carrying females fetuses were euthanized at 199 and 268d of gestation (DG; n=3 for H or M on each DG). Fifteen cows from H (n=6) and M intake (n=9) carrying male fetuses were euthanized at 139, 199, and 241 DG (n=2 for H and n=3 for M on each DG). Fetal gonads and pituitary gland were sampled for gene expression and histological analyses. Sex-specific responses to maternal intake were observed. Primordial and total follicle numbers were lower in fetal ovaries from H than in M intake cows. These results were the reverse for preantral and antral follicles. Volumetric proportion and diameter of seminiferous cord were lower in fetal testis of H than M intake cows. The expression level of FSHB was greater in pituitary gland of the female fetus from H compared with M intake cows, irrespective of DG, whereas LHB gene expression did not differ. In males, FSHB and LHB gene expression levels were similar between maternal intake groups. Fetal ovarian expression of P450 aromatase, StAR, BMPR2, TGFBR1, GDF9, FSHR, Bax, and CASP3 genes were higher in H than in M intake cows, irrespective of DG. Fetal testicular expression of StAR, HSD17B3, IGF1, IGF2, and IGF1R genes was higher in M than in H intake cows. The differences in gene expression for steroidogenesis, folliculogenesis, and apoptosis may explain the distinct pattern of follicular growth between offspring of M and H intake cows. By contrast, the lower volumetric proportion, diameter, and length of seminiferous cord may relate to decreased gene expression in fetal testis from H intake cows. In conclusion, maternal H intake seems to affect fetal ovarian follicular growth and number of follicles, which may affect the size of ovarian reserve in their offspring. In male fetus, maternal H intake seems to disturb testicular development and may have implications on sperm production. The underlying mechanism of differential gene expression and the effect on offspring reproductive potential should be the focus of further research, especially considering larger sample size, reducing the chance for type I errors.

Desempenho produtivo de bovinos alimentados com cana-de-açúcar com diferentes níveis de concentrado / Productive performance of cattle fed sugarcane with different concentrate levels

Foram avaliados o consumo, as digestibilidades totais e ruminais e as taxas de digestão (kd) e de passagem (kp) ruminal dos nutrientes de dietas constituídas de cana-de-açúcar in naturae diferentes níveis de concentrado. Utilizaram-se cinco bovinos mestiços, fistulados no rúmen, com peso corporal inicial de 300±50kg, distribuídos em delineamento em quadrado latino 5x5. As dietas experimentais foram constituídas de: 1) 100% cana-de-açúcar in natura(CA); 2) 80% de CA + 20% de concentrado (C); 3) 60% de CA + 40% de C; 4) 40% de CA + 60% de C; e 5) 20% de CA + 80% de C. Os dados foram analisados utilizando-se o procedimento MIXED do SAS (versão 9.1), bem como análise de regressão e 5% como nível crítico de probabilidade para o erro tipo I. O consumo de matéria seca (MS), expresso em kg/dia ou g/kg de peso corporal foi influenciado (P<0,05) pelos níveis de concentrado. Os demais consumos calculados em kg/dia também foram influenciados (P<0,05) pelos níveis de concentrado, assim como os consumos de FDNcp, FDNi e NDT, expressos em g/kg de peso corporal. O pool (kg/dia) foi aumentado linearmente à medida que se elevaram os níveis de concentrado para MS. Comportamento similar foi observado para ki (h-1) da MS e da FDNcp. As taxas de digestão (kdh-1) da MS, da MO e da FDNcp aumentaram linearmente com a elevação dos níveis de concentrado na dieta. Estimaram-se máximos fluxos omasais (kg/dia) com 52,22 e 31,45% de concentrado na dieta, respectivamente, para MS e FDNi. A digestibilidade aparente (DA) da MS também foi influenciada pelos níveis de concentrado na dieta. As digestibilidades ruminais da MS e da FDN aumentaram linearmente (P<0,05) com a adição de níveis crescentes de concentrado. Conclui-se que níveis próximos de 65% de concentrado resultam em maior ingestão de energia em dietas contendo cana-de-açúcar.

An experiment was conducted to evaluate the feed intake, total and ruminal dietary component digestibility, ruminal digestion (kd) and passage rates (kp) of cattle fed sugarcane with different levels of concentrate. Five crossbred (Holstein x Zebu) with ruminal cannulas and initial body weight of 300±50kg were used in this experiment. Steers were assigned into a 5x5 Latin Square design balanced for residual effects. The experimental periods were divided into 16 d with 8 d for adaptation and 8 d for sample collection. Four treatments were used: 1) 100% fresh sugarcane (SC); 2) 80% SC + 20% concentrate (C); 3) 60% SC + 40% C; 4) 40% SC + 60% C; 5) 20% SC + 80% C. Urea/ammonium sulfate was used at 9:1 ratio to correct the protein level of sugarcane. A regression analysis was performed using the MIXED procedure. Dry matter intake (DMI) expressed either as kg/day or g/kg of BW was affected (P<0.05) by concentrate level and the maximum intake was estimated to be 63.61 and 75.04% of concentrate respectively. Intake (kg/day) of OM, CP, NDFap, TDN, iNDF, digestible NDFap (NDFapD), and NFC were affected by concentrate level (P<0.05) and their maximum intake was estimated to be 63.21; 63.78; 53.14; 64.31; 32.49; 88.47 e 55.46% of concentrate respectively. The ruminal pool size of DM and CP as well as ki (h-1) DM, CP, OM, NDFap, and NFC increased linearly as the dietary levels of concentrate increased. The ruminal digestion rate (kg h-1) of DM and NDFap as well as the passage rate (kg h-1) of CP increased linearly as the dietary levels of concentrate increased. The maximum omasal flux (kg/day) of DM and iNDF were estimated to be 52.22 and 31.45% of dietary concentrate respectively. Apparent digestibility of DM, EE, and NFC also increased linearly (P<0.05) with the increase of concentrate levels. Data suggest that levels up to 65% of concentrate result in greater energy intake by cattle fed sugarcane based diets.

Effects of day of gestation and feeding regimen in Holstein × Gyr cows: II. Maternal and fetal visceral organ mass.

This study investigated the influence of day of gestation (DG) and feeding regimens (FR) on maternal and fetal visceral organ mass in Holstein × Gyr cows. Forty-four pregnant multiparous Holstein × Gyr cows with an average initial body weight of 480±10.1 kg and an average initial age of 5±0.5 yr were allocated to 1 of 2 FR: ad libitum (AL; n=20) or maintenance level (ML; n=24). Maintenance level was considered to be 1.15% of body weight (dry matter basis) and met 100% of the energy requirements; AL provided 190% of the total net energy requirements. Cows were individually fed a corn silage and concentrate-based diet composed of 93% roughage and 7% concentrate (dry matter basis) as a total mixed ration twice daily. Pregnant cows were slaughtered at 4 DG: 139 (n=11), 199 (n=11), 241 (n=11), and 268 (n=11) d, which was followed by necropsy. Mass of heart, liver, and gastrointestinal tract was heavier in AL- than in ML-fed cows. Mammary gland mass was heavier in AL- than in ML-fed cows, and the heaviest mass was observed at 268 d of gestation. Feeding regimen did not influence fetal body weight in this study. The majority of the visceral organ masses were similar in fetuses from cows fed AL or ML. These data indicate that maternal feed restriction does not affect the development of most fetal organs or fetal development; however, some maternal organs are affected by the FR provided. Moreover, the negative effect on mammary gland mass caused by ML feeding will probably not affect the subsequent lactation because the crude protein concentration in the mammary gland increased with ML feeding. However, we suggest that the AL diet in pregnant dry cows should be provided with caution because the amount of fat in the mammary gland increased at 268 d of gestation.

Effects of day of gestation and feeding regimen in Holstein × Gyr cows: I. Apparent total-tract digestibility, nitrogen balance, and fat deposition.

This study investigated how feeding regimen (FR) alters apparent total-tract digestibility, performance, N balance, excretion of purine derivatives, and fat deposition in Holstein × Gyr cows at different days of gestation (DG). Forty-four pregnant multiparous Holstein × Gyr cows with an average initial body weight of 480±10.1 kg and an initial age of 5±0.5 yr old were allocated to 1 of 2 FR: ad libitum (AL; n=20) and maintenance level (ML; n=24). Maintenance level was considered to be 1.15% of body weight on a dry matter (DM) basis and met 100% of the energy requirements, whereas AL provided 190% of total net energy requirements. Data for hot and cold carcass dressing, fat deposition, average daily gain, empty body gain, and average daily gain without the gravid uterus were analyzed as a 4×2 factorial design. Intake, apparent total-tract digestibility, N balance, urinary concentration of urea, and purine derivatives data were analyzed as repeated measurements taken over the 28-d period (122, 150, 178, 206, 234, and 262 d of gestation). Cows were individually fed a corn silage-concentrate based diet composed of 93% roughage and 7% concentrate (DM basis) as a total mixed ration. Pregnant cows were slaughtered on 4 different DG: 139 (n=11), 199 (n=11), 241 (n=11), and 268 d (n=11). Overall, DM intake decreased as DG increased. This decrease observed in DM intake may be associated with the reduction in ruminal volume caused by the rapid increase in fetal size during late gestation. We observed an interaction for DM and organic matter apparent total-tract digestibility between FR and DG; at 150, 178, and 206 d of gestation, ML-fed cows had greater DM and organic matter apparent total-tract digestibility values than AL-fed cows. Rib fat thickness, mesentery, and kidney, pelvic, and heart fat were greater in AL-fed than in ML-fed cows at all DG, with the exception of rib fat thickness on d 139. Ad libitum-fed cows excreted more N in their feces and urine compared with ML-fed cows. Pregnant cows that were fed at maintenance had greater digestibility during some DG, excreted less N in feces and less N and urea in urine, and deposited less fat in the body. We therefore recommend ML (1.15% of body weight with 93% of roughage) as a FR for pregnant dry cows; however, during the last month of gestation, AL seems to be the most appropriate FR to avoid loss of body weight.

Effects of day of gestation and feeding regimen in Holstein × Gyr cows: III. Placental adaptations and placentome gene expression.

This study investigated the influence of day of gestation (DG) and feeding regimens (FR) on the expression of genes responsible for placenta development, nutrient transfer, and angiogenic factors in Holstein × Gyr cows. Forty pregnant multiparous Holstein × Gyr cows with an average initial body weight of 482±10.8kg and an initial age of 5±0.8 yr were allocated to 1 of 2 FR: ad libitum (AL; n=20) or maintenance level (ML; n=20). Maintenance level was considered to be 1.15% of body weight (dry matter basis) and met 100% of the net energy requirements and AL provided 190% of the total net energy requirements. Cows were slaughtered at 4 DG: 139, 199, 241, and 268d. After the cows were slaughtered, the placenta and uterus were separated and weighed. Caruncles and cotyledons were individually separated, counted, and weighed. Placenta expressed as kilograms and grams per kilogram of empty body weight (EBW) was heavier in ML- than in AL-fed cows at 268d of gestation. Placenta expressed as kilograms and grams per kilogram of EBW was the lightest at 139d of gestation, and the greatest mass was observed at 268d in ML-fed cows. However, in AL-fed cows, the heaviest placenta expressed as grams per kilogram of EBW was observed from 199d of gestation. Placentomes expressed as grams per kilogram of EBW were heavier in ML-fed cows during gestation, and the number of placentomes was greater in ML-fed cows at 268d of gestation. We observed that IGFR1 and IGFR2 were involved in placenta adaptations when ML was provided, as their expression in placentome cells was greater in ML-fed cows at 268d of gestation. The genes responsible for angiogenesis were also greater in ML-fed cows: VEGFA, GUCY1B3, HIFA, FGF2, and NOS3 were altered by FR and DG interaction and they were greater in ML-fed cows at 268d of gestation. In addition, VEGFB and ANGPT2 did not show interactions between FR and DG, but they were greater in ML-fed cows. Thus, we suggest that the placenta from an ML-fed cow develops adaptations to the reduced nutrient supply by altering its structure and gene expression, thereby developing mechanisms for potential increased nutrient transfer efficiency to the fetus.

Digesta sampling sites and marker methods for estimation of ruminal outflow in bulls fed different proportions of corn silage or sugarcane.

The objective of this experiment was to assess ruminal outflow and apparent total-tract digestibility using digesta samples from 3 sites (reticulum, omasum, and abomasum) and 3 marker methods (single marker: indigestible NDF [iNDF; sample without separation]; double marker: iNDF + Co-EDTA [filtered sample]; and triple marker: iNDF + ytterbium [Yb] acetate + Co-EDTA [filtered and centrifuged]) in bulls fed corn silage and sugarcane-based diets. Eight crossbred (Holstein × Zebu) bulls (353 ± 37 kg of BW; 24 ± 1 mo of age) with ruminal and abomasal cannulas were randomly distributed into two 4 × 4 Latin squares that were balanced for residual effects. The following experimental diets were used: 1) 60% corn silage + 40% concentrate, 2) 40% corn silage + 60% concentrate, 3) 60% fresh sugarcane + 40% concentrate, and 4) 40% fresh sugarcane + 60% concentrate. Reticular, omasal, and abomasal digesta samples were collected at 9-h intervals over 3 d. At the end of the experiment, a composite sample was prepared for each bull, and these samples were subsequently assigned to the 3 marker methods. The concentrations of CP, NDF, and iNDF of reticular digesta differed (P < 0.01) from those of the omasum and abomasum. Use of omasal and abomasal samples led to similar estimates of ruminal outflow and ruminal digestibility for DM (P = 0.65), OM (P = 0.68), CP (P = 0.85), and NDF (P = 0.57). In contrast, the ruminal outflow of digesta based on reticular sampling appeared to be underestimated. We recommend sampling from the omasum because sampling from this region is less invasive than sampling from the abomasum. Although we did not observe differences in ruminal NDF digestibility among the different marker methods, we did observe that ruminal digestibility of CP was greater for the single marker method than for the double and triple marker methods; we therefore recommend the use of the double or triple marker method.

The impact of dietary sugarcane addition to finishing diets on performance, apparent digestibility, and fatty acid composition of Holstein × Zebu bulls.

The objective of this study was to evaluate the influence of corn silage (CS) or sugarcane (SC) additions to finishing diets on voluntary intake, apparent digestibility, animal performance, and LM fatty acid (FA) composition in crossbred Holstein × Zebu bulls. Forty-two bulls (initial BW 328 ± 39.2 kg and an initial age of 23.5 mo) were used in this experiment. A completely randomized design was used to examine roughage source (CS vs. SC) and ratio of concentrate (CO) on nutrient utilization and production. The experimental diets consisted of 1) 60% CS and 40% CO on DM basis fed for 84 d (CS60:40), 2) 60% CS and 40% CO on DM basis fed for 42 d and 40% CS and 60% CO on DM basis fed for 42 d (CS reverse diet [CSR]), 3) 40% CS and 60% CO on DM basis fed for 84 d (CS40:60), 4) 60% SC and 40% CO on DM basis fed for 84 d (SC60:40), 5) 60% SC and 40% CO on DM basis fed for 42 d and 40% SC and 60% CO on DM basis fed for 42 d (SC reverse diet [SCR]), and 6) 40% SC and 60% CO on DM basis fed for 84 d (SC40:60). Dry matter and NDF intakes were greater (P < 0.01) in bulls fed CS-based diets than bulls fed SC-based diets. The greatest (P < 0.01) DM and CP digestibility were observed in bulls fed SC-based diets. Bulls fed CSR and CS40:60 had greater (P < 0.01) ADG than bulls fed SC-based diets. However, ADG was similar (P = 0.52) in bulls fed CS60:40, CSR, and SC40:60 diets. The percentage of C18:3 n-3 and C20:5 n-3 were greater (P < 0.01) in LM of bulls fed SC-based diets. The percentage of CLA was greater (P < 0.01) in LM of bulls fed SC60:40 than those fed CS-based diets. The findings of the present study indicate that SC40:60 can replace CS60:40 and CSR in finishing diets and, moreover, roughage source significantly altered the FA composition of crossbred Holstein × Zebu bulls LM.

Development of equations to estimate microbial contamination in ruminal incubation residues of forage produced under tropical conditions using 15N as a label.

The objective of this study was to use (15)N to label microbial cells to allow development of equations for estimating the microbial contamination in ruminal in situ incubation residues of forage produced under tropical conditions. A total of 24 tropical forages were ruminal incubated in 3 steers at 3 separate times. To determine microbial contamination of the incubated residues, ruminal bacteria were labeled with (15)N by continuous intraruminal infusion 60 h before the first incubation and continued until the last day of incubation. Ruminal digesta was collected for the isolation of bacteria before the first infusion of (15)N on adaptation period and after the infusion of (15)N on collection period. To determine the microbial contamination of CP fractions, restricted models were compared with the full model using the model identity test. A value of the corrected fraction "A" was estimated from the corresponding noncorrected fraction by this equation: Corrected "A" fraction (A(CP)C) = 1.99286 + 0.98256 × A" fraction without correction (A(CP)WC). The corrected fraction "B" was estimated from the corresponding noncorrected fraction and from CP, NDF, neutral detergent insoluble protein (NDIP), and indigestible NDF (iNDF) using the equation corrected "B" fraction (B(CP)C) = -17.2181 - 0.0344 × fraction "B" without correction (B(CP)WC) + 0.65433 × CP + 1.03787 × NDF + 2.66010 × NDIP - 0.85979 × iNDF. The corrected degradation rate of "B" fraction (kd)was estimated using the equation corrected degradation rate of "B" fraction (kd(CP)C) = 0.04667 + 0.35139 × degradation rate of "B" fraction without correction (kd(CP)WC) + 0.0020 × CP - 0.00055839 × NDF - 0.00336 × NDIP + 0.00075089 × iNDF. This equation was obtained to estimate the contamination using CP of the feeds: %C = 79.21 × (1 - e(-0.0555t)) × e(-0.0874CP). It was concluded that A and B fractions and kd of CP could be highly biased by microbial CP contamination, and therefore these corrected values could be obtained mathematically, replacing the use of microbial markers. The percentage of contamination and the corrected apparent degradability of CP could be obtained from values of CP and time of incubation for each feed, which could reduce cost and labor involved when using (15)N.