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.

3D printing in the design of pharmaceutical dosage forms.

Three-dimensional (3D) printing technologies are manufacturing approaches with widespread use in industry (e.g. automotive, automobile, pharmaceutical industries). With regard to its use in pharmaceutical industry, 3D printing is demonstrating to be of added value attributed to the possibility of printing tailored pharmaceutical products, namely personalized medical devices, such as implants and other dosage forms. However, with the approval of the first 3D-printed drug-product in 2015, a new perspective has arisen, i.e. the use of this technology to produce solid oral dosage forms exhibiting complex drug release profiles and allowing for individual dosing. Technological hurdles and regulatory issues still have to be overcome before this technology can truly find its place in the healthcare sector, where it can certainly contribute to a personalized and patient-centered healthcare. This manuscript offers a comprehensive analysis of the most extensively used methods of 3D printing in the pharmaceutical field, with examples of solid oral dosage forms and other medical devices currently under development or already marketed.

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.

Determination of energy and protein requirements for crossbred Holstein × Gyr preweaned dairy calves.

The objective was to quantify the energy and protein nutritional requirements of Holstein × Gyr crossbred preweaned dairy calves until 64 d of age. Thirty-nine Holstein × Gyr crossbred male calves with an average initial live weight (mean ± SEM; for all next values) of 36 ± 1.0 kg were used. Five calves were slaughtered at 4 d of life to estimate the animals' initial body composition (reference group). The remaining 34 calves were distributed in a completely randomized design in a 3 × 2 factorial arrangement consisting of 3 levels of milk (2, 4, or 8 L/d) and 2 levels of starter feed (presence or absence in diet). At 15 and 45 d of life, 4 animals from each treatment were subjected to digestibility trials with total collection of feces (for 72 h) and urine (for 24 h). At 64 d of age, all animals were slaughtered, their gastro-intestinal tract was washed to determine the empty body weight (EBW; kg), and their body tissues were sampled for subsequent analyses. The net energy requirement for maintenance was estimated using an exponential regression between metabolizable energy intake and heat production (both in Mcal/EBW0.75 per d) and was 74.3 ± 5.7 kcal/EBW0.75 per d, and was not affected by inclusion of starter feed in the diet. The metabolizable energy requirement for maintenance was determined at the point of zero energy retention in the body and was 105.2 ± 5.8 kcal/EBW0.75 per d. The net energy for gain was estimated using the EBW and the empty body gain (EBG; kg/d) as 0.0882 ± 0.0028 × EBW0.75 × EBG0.9050±0.0706. The metabolizable energy efficiency for gain (kg) of the milk was 57.4 ± 3.45%, and the kg of the starter feed was 39.3 ± 2.09%. The metabolizable protein requirement for maintenance was 3.52 ± 0.34 g/BW0.75 per d. The net protein required for each kilogram gained was estimated as 119.1 ± 32.9 × EBW0.0663±0.059. The metabolizable protein efficiency for gain was 77 ± 8.5% and was not affected by inclusion of starter feed in the diet. In conclusion, the energy efficiency for gain of milk is higher than that of starter and the net protein required per unit protein gain increases with empty body weight.

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.

Dissimilarity between Mouriri elliptica (Mart.) plants cultivated in vitro and in situ through anatomic parameters.

Mouriri elliptica (Mart.) is a genetic resource of the Cerrado domain, which has potential for food and medicinal use. A few studies have been performed on its in vitro propagation, and no studies have examined dissimilarities between plants of this species when cultivated in situ or in vitro. Therefore, the objective of the present study was to identify in vitro cultivation conditions that permit the formation of plantlets with leaf anatomical features that are less dissimilar to those of plants grown in situ. Thus, an anatomical study of the leaves was conducted to investigate the adaxial epidermis thickness, abaxial epidermis thickness, chlorenchyma thickness, stomatal crypt depth, stomatal crypt density, and leaf surface stomatal crypt aperture area. The distance between phenotypes was determined based on micromorphometric data, and unweighted pair group mean arithmetic clustering was performed. Four different groups were tested, and cultivation with sucrose and 50 and 75 µmol⋅m-2⋅s-1 irradiance was found to promote plantlet development and maximize similarity to plants cultivated in situ. The most important anatomical parameters in this study were the stomatal crypt aperture area and the crypt density. This study is of importance for the anatomical characterization of M. elliptica (Mart.) leaves, as it identifies plasticity as a function of in vitro culture conditions.

Effects of Supplements with Different Protein Contents on Nutritional Performance of Grazing Cattle During the Rainy Season.

The objective of this study was to evaluate the effects of supplements with different crude protein (CP) contents on grazing cattle intake, digestibility, ruminal fermentation pattern, and nitrogen (N) metabolism characteristics during the rainy season. Five ruminal and abomasal cannulated Holstein×Zebu steers (296 kg body weight, BW) were used in a 5×5 Latin square design. The animals grazed five signal grass paddocks (0.34 ha). The five treatments evaluated were: Control (no supplement) and 1.0 g of supplement/kg BW with 0, 330, 660, and 1,000 g of CP/kg as-fed. The supplement was composed of starch, soybean meal, urea, and ammonium sulphate. There was a positive linear effect (p≤0.033) of the CP content in the supplements on the organic matter (OM), CP, and digested OM intakes. The provision of supplements did not increase (p≥0.158), on average, total and ruminal digestibilities of OM and CP. However, the increase in CP content in the supplements caused a positive linear effect (p≤0.018) on ruminal digestibilities of OM and CP. Additionally, a quadratic effect of the CP contents of the supplements were observed (p = 0.041) for the ruminal digestibility of neutral detergent fiber corrected for ash and protein, with the highest estimate obtained with the CP content of 670 g/kg. The supply of supplements increased (p<0.001) the ruminal ammonia N concentration, which also changed linearly and positively (p<0.001) according to increase in CP content in the supplements. The apparent N balance and relative N balance (g/g N intake) were not, on average, changed (p≥0.164) by the supplements supply. However, both showed a tendency of a linear increase (p≤0.099) with increasing supplement CP content. The supplements increased (p = 0.007) microbial N production in the rumen, which also changed linearly and positively (p = 0.016) with increasing supplement CP content. In conclusion, protein supplementation in grazing cattle during the rainy season, while stimulating voluntary forage intake, results in higher efficiency of N utilization when compared to energy supplementation. This is a possible response to increased microbial protein synthesis in the rumen and improved N status in the animal body.

Effects of raw milk and starter feed on intake and body composition of Holstein × Gyr male calves up to 64 days of age.

The objective was to evaluate the effect of supplying different levels of raw milk, alone or in combination, with access to a starter feed, on the intake, digestibility, daily gain, N balance, and body composition of Holstein × Gyr crossbred suckling calves until 64 d of age. Thirty-nine male calves aged 4 d with an average initial live weight of 36 ± 1.0 kg were used. Five calves were defined as a reference group and slaughtered at 4 d of age to estimate the initial body composition of the animals. The other calves were distributed according to a completely randomized design in a 3 × 2 factorial arrangement consisting of 3 levels of milk (2, 4, or 8 L/d) and 2 levels of starter feed (presence or absence in diet). At 15 and 45 d of age, 4 animals from each treatment were subjected to digestibility trials with total collection of feces and urine and sampling of feeds. At 64 d of age, all animals were slaughtered and their body tissues were sampled for analyses. Total dry matter and nutrient intake increased linearly and starter intake decreased linearly in response to the supply of increasing amounts of milk. The digestibility coefficient of organic matter was not affected by the inclusion of starter feed and increased linearly as milk supply was elevated. Daily gain was greater at increased milk supply levels and also greater when starter was supplied, without any interaction between milk supply level and the presence or absence of starter. Fecal N excretion and N retention were higher in the animals fed starter feed. Fecal N excretion was not affected by milk levels, whereas N retention was affected. Body protein and ash contents decreased linearly according to increased milk allowance. In contrast, fat body content increased linearly according to milk supply. The presence of starter feed in the diet was responsible for the increased body fat content, but had no effect on protein or ash content. In conclusion, weight gain and N retention in calves up to 64 d of age increased with milk supply. Ad libitum access to starter further increased weight gain and N retention and no interaction with milk allowance level occurred. However, greater levels of milk are also associated with reduced starter feed intake, in addition to increased body fat content.

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.

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.

Evaluation of Acid Digestion Procedures to Estimate Mineral Contents in Materials from Animal Trials.

Rigorously standardized laboratory protocols are essential for meaningful comparison of data from multiple sites. Considering that interactions of minerals with organic matrices may vary depending on the material nature, there could be peculiar demands for each material with respect to digestion procedure. Acid digestion procedures were evaluated using different nitric to perchloric acid ratios and one- or two-step digestion to estimate the concentration of calcium, phosphorus, magnesium, and zinc in samples of carcass, bone, excreta, concentrate, forage, and feces. Six procedures were evaluated: ratio of nitric to perchloric acid at 2:1, 3:1, and 4:1 v/v in a one- or two-step digestion. There were no direct or interaction effects (p>0.01) of nitric to perchloric acid ratio or number of digestion steps on magnesium and zinc contents. Calcium and phosphorus contents presented a significant (p<0.01) interaction between sample type and nitric to perchloric acid ratio. Digestion solution of 2:1 v/v provided greater (p<0.01) recovery of calcium and phosphorus from bone samples than 3:1 and 4:1 v/v ratio. Different acid ratios did not affect (p>0.01) calcium or phosphorus contents in carcass, excreta, concentrate, forage, and feces. Number of digestion steps did not affect mineral content (p>0.01). Estimated concentration of calcium, phosphorus, magnesium, and zinc in carcass, excreta, concentrated, forage, and feces samples can be performed using digestion solution of nitric to perchloric acid 4:1 v/v in a one-step digestion. However, samples of bones demand a stronger digestion solution to analyze the mineral contents, which is represented by an increased proportion of perchloric acid, being recommended a digestion solution of nitric to perchloric acid 2:1 v/v in a one-step digestion.

Grazing behavior and locomotion of young bulls receiving different nutritional plans in a tropical pasture.

The objectives of this study were to compare visual observation and an electronic grazing time method and to evaluate the effects of nutritional plans on intake, grazing behavior and horizontal and vertical locomotion of young bulls in a tropical pasture. Thirty-nine Nellore young bulls with an average body weight of 345±9.3 kg kept in pasture were used. The experimental treatments consisted of: restricted: animals kept in a plot with a low mass of forage receiving mineral mixture only; control: animals receiving mineral mixture only; HPHC: a high protein and high carbohydrate supplement; HPLC: a high protein and low carbohydrate supplement; LPHC: a low protein and high carbohydrate supplement; LPLC: a low protein and low carbohydrate supplement. GPS collars equipped with activity sensors were used. Information about head position, latitude, longitude and altitude were recorded. Daytime grazing behavioral patterns monitored by a continuous focal animal recording method was compared to behavior estimated by the activity sensor. Feed intake was estimated by a marker method. The Restricted group presented lower (p<0.05) intake of dry matter and TDN. However, difference in dry matter intake was not found (p>0.05) between non-supplemented and supplemented animals. Difference was not found (p>0.05) in daytime grazing time obtained by visual observation or the activity sensor method. The restricted group showed longer (p<0.05) grazing time (9.58 h/d) than other groups, but difference was not found (p>0.05) in the grazing time between Control (8.35 h/d) and supplemented animals (8.03 h/d). The Restricted group presented lower (p<0.05) horizontal locomotion distance (2,168 m/d) in comparison to other groups (2,580.6 m/d). It can be concluded that the use of activity sensor methods can be recommended due to their being similar to visual observation and able to record 24-h/d. While supplements with high carbohydrates reduce pasture intake, they do not change grazing behavior. Moderate supplementation (until 50% of protein requirement and 30% of energy requirement) of beef cattle on tropical pasture has no effect on daily locomotion.

Biomarkers of pollution by glyphosate in the lichens, Parmotrema tinctorium and Usnea barbata.

Glyphosate is a herbicide commonly used in agriculture for weed control. Current agricultural production demands vast amounts of this product, which are applied by ground or aerial spraying. The concomitant aerial currents promote glyphosate drift to vegetated or urban areas. In this context, we hypothesized that the lichens, Parmotrema tinctorum and Usnea barbata, could be sensitive to the action of glyphosate and therefore be used to bio-indicate the presence of this herbicide in areas affected by drift. Since living organisms respond in different ways to the action of herbicides, our interest was also to indicate biological markers responsive to the action of glyphosate, through concentrations and exposure times of the thallus, besides identifying the most sensitive species. We evaluated the effect of different concentrations (0.0, 4.8, 9.6, and 19.2 mg L-1) and exposure times (24, 48, and 72 hours) to glyphosate on the morphoanatomy, photobiont vitality, photosynthetic efficiency, and oxidative metabolism of the thalli. We found that the lichens, P. tinctorum and U. barbata, respond to glyphosate stress, with prospects for use in the biomonitoring of pollutant dispersal from plantation areas. When using P. tinctorum as a bioindicator, lichen morphoanatomy, photobiont vitality, and photosynthetic pigment concentration were efficient biomarkers for the effect of concentration and exposure time. For U. barbata, the lichenic morphoanatomy and the activity of SOD and APX enzymes were essential tools to indicate the herbicide action. Parmotrema tinctotum, however, was characterized as more sensitive in bio-indicating the presence of this herbicide to diagnose the air quality in urban areas or vegetation sectors adjacent to agricultural environments.

Development of equations to predict carcass weight, empty body gain, and retained energy of Zebu beef cattle.

The accurate supply of energy is essential to optimize livestock productivity and profitability. Furthermore, replacing empty BW gain (EBG) with carcass gain (CG) might be a suitable alternative to estimate the retained energy (RE) of beef cattle. Thus, this multi-analysis study was conducted aiming to estimate and validate new equations to predict carcass weight (CW), EBG, and RE of Zebu, beef crossbred, and dairy crossbred. A database composed by 1112 animals encompassing bulls, steers, heifers of different genetic groups (Zebu, beef crossbred, and dairy crossbred), and two types of slaughter plants (commercial and experimental) was used for generating the new CW equation. For the development of the EBG and RE equations, a database of 636 observations composed of bulls, steers, and heifers of different genetic groups (Zebu, beef crossbred, and dairy crossbred) was assembled. The validation of new equations was performed using independent databases composed by 137 observations (80 for CW and 57 for EBG and RE). The new approaches for EBG and RE validation also included data from our research group studies (Inside) and independent data from literature publications (Outside). Furthermore, the new RE equation was compared to the current model devised by the nutritional requirements, diet formulation, and performance prediction of Zebu and crossbred cattle (BR-CORTE, 2016). Validation analyses were performed by using the Model Evaluation System (MES; 3.1.13, College Station, US). The CW was accurately estimated by the new equation when using both commercial and experimental data. Also, the equations developed in this study accurately estimated EBG and RE using both inside and outside data. In conclusion, equations proposed in this study accurately and precisely estimated CW, EBG, and RE of Zebu beef cattle that composed validation data set. Therefore, we suggest the following equations to estimate CW, EBG, and RE of Zebu cattle: CW, kg = -11.0±1.56 + P + ((0.609±0.005 + G + B) × SBW); EBG (kg) = 0.044±0.017 + 1.47±0.026 × CG; RE (MJ/d) = 4.184 × (0.082±0.002 × EQEBW0.75 × CG0.777±0.039), where P = slaughter plant effect, if commercial = -10.98, if experimental =0; G = gender effect, if steer = 0, if bull = 0.008169 and if heifer = -0.00612; B = genotype effect, if Zebu = 0, if dairy crossbreds = -0.03301 and if beef crossbreds = -0.01595; SBW = shrunk BW; CG = carcass gain; EQEBW = equivalent empty BW.

Metabolizable amino acids and energy requirements of Nellore and crossbred Angus × Nellore bulls fed rations of different crude protein concentrations.

Growth rate of cattle depends on their genetic makeup and nutrient intake. Moreover, increased growth rate may lead to increased amino acid (AA) requirements. Therefore, we evaluated the AA content of the empty body and estimated the net AA and energy requirements of purebred and crossbred beef bulls fed rations of different dietary CP concentrations. We performed a comparative slaughter experiment with 24 Nellore and 24 Angus × Nellore (A × N) bulls (8 months; initial shrunk BW: Nellore = 208.0 ±â€¯12.78 kg; A × N = 221.9 ±â€¯14.16 kg). Eight bulls (four Nellore and four A × N) were designated as the reference group, eight bulls (four Nellore and four A × N) were fed to maintenance level and 32 bulls (16 Nellore and 16 A × N) were fed ad libitum. The 32 bulls fed ad libitum were distributed using a completely randomized design in a 2 × 3 factorial scheme with two genetic groups (Nellore or A × N) and three dietary CP contents (100, 120 or 140 g CP/kg DM), being four groups with five bulls and two groups with six bulls. The experimental period lasted for 224 days. There were no interactions (P ≥ 0.056) between the dietary CP contents and genetic groups for any of the response variables. The dietary CP contents did not affect (P ≥ 0.062) the AA content in the empty body (g/kg empty BW [EBW]), with exception for Tryptophan (P = 0.027, linear effect). The dietary CP contents did not affect (P ≥ 0.051) AA content in the empty body (g/100 g of CP), with exception for Alanine (P = 0.013) that responded quadratically to dietary CP increase. The equations to estimate the net Lysine (Lys) and Methionine (Met) requirements (g/100 g of CP) were: Lys = 5.1 × EBW0.0594 and Met = 1.7 × EBW0.0255. Metabolizable Lys and Met to metabolizable energy (ME) ratios decreased as bulls EBW increased. Also, the metabolizable protein to ME ratio decreased as bulls EBW increased. In conclusion, the present study provides useful information regarding net and metabolizable requirements of AA of purebred and crossbred beef bulls. In the future, after the validation of the equations, these results can be used to calculate the AA requirements for growth of purebred and crossbred beef bulls. Nevertheless, it is important to highlight that the small sample size was one limitation of this present experiment.

Predicting enteric methane production from cattle in the tropics.

Accurate estimates of methane (CH4) production by cattle in different contexts are essential to developing mitigation strategies in different regions. We aimed to: (i) compile a database of CH4 emissions from Brazilian cattle studies, (ii) evaluate prediction precision and accuracy of extant proposed equations for cattle and (iii) develop specialized equations for predicting CH4 emissions from cattle in tropical conditions. Data of nutrient intake, diet composition and CH4 emissions were compiled from in vivo studies using open-circuit respiratory chambers, SF6 technique or the GreenFeed® system. A final dataset containing intake, diet composition, digestibility and CH4 emissions (677 individual animal observations, 40 treatment means) obtained from 38 studies conducted in Brazil was used. The dataset was divided into three groups: all animals (GEN), lactating dairy cows (LAC) and growing cattle and non-lactating dairy cows (GCNL). A total of 54 prediction equations available in the literature were evaluated. A total of 96 multiple linear models were developed for predicting CH4 production (MJ/day). The predictor variables were DM intake (DMI), gross energy (GE) intake, BW, DMI as proportion of BW, NDF concentration, ether extract (EE) concentration, dietary proportion of concentrate and GE digestibility. Model selection criteria were significance (P < 0.05) and variance inflation factor lower than three for all predictors. Each model performance was evaluated by leave-one-out cross-validation. The Intergovernmental Panel on Climate Change (2006) Tier 2 method performed better for GEN and GCNL than LAC and overpredicted CH4 production for all datasets. Increasing complexity of the newly developed models resulted in greater performance. The GCNL had a greater number of equations with expanded possibilities to correct for diet characteristics such as EE and NDF concentrations and dietary proportion of concentrate. For the LAC dataset, equations based on intake and animal characteristics were developed. The equations developed in the present study can be useful for accurate and precise estimation of CH4 emissions from cattle in tropical conditions. These equations could improve accuracy of greenhouse gas inventories for tropical countries. The results provide a better understanding of the dietary and animal characteristics that influence the production of enteric CH4 in tropical production systems.

Effects of soybean oil supplementation on performance, digestion and metabolism of early lactation dairy cows fed sugarcane-based diets.

Sugarcane is an important forage source for dairy cows in tropical countries. However, it provides limited digestible fiber and energy intake, and fat supplementation can be a way to increase energy density and decrease dietary, non-fiber carbohydrates concentrations. We aimed to evaluate the performance, digestion and metabolism of dairy cows in early lactation fed different concentrations of soybean oil (SBO) in sugarcane-based diets. Fourteen primiparous (545±17.2 kg of BW) and eight multiparous (629±26.7 kg BW) Holstein dairy cows were used according to a randomized block design. After calving, diets were randomly assigned to cows within the two parity groups. Diets were formulated with increasing concentrations of SBO (g/kg dry matter (DM)): control (0), low (LSBO; 15.7), medium (MSBO; 44.3) and high (HSBO; 73.4). The study was performed from calving until 84 days in milk, divided into three periods of 28 days each. Dry matter intake (DMI) was affected quadratically in response to SBO addition with the greatest and lowest values of 19.0 and 16.0 kg/day for LSBO and HSBO diets, respectively. The digestibility of potentially digestible NDF was quadratically affected by SBO with the greatest value of 623 g/kg for LSBO diet. Both milk and energy-corrected milk (ECM) production were quadratically affected by SBO inclusion, with greatest ECM values of 27.9 and 27.3 for LSBO and MSBO, respectively. Soybean oil inclusion linearly decreased milk fat concentration by 13.2% from control to HSBO. The CLA t10,c12-18:2 was observed in milk fat only for MSBO and HSBO diets. Soybean oil inclusion did not affect plasma glucose or serum concentrations of total proteins, globulins, albumin, urea nitrogen, beta-hydroxybutyrate, non-esterified fatty acids or insulin. Serum concentrations of total cholesterol, triglycerides and low-density lipoprotein increased with SBO supplementation. Soybean oil inclusion in sugarcane-based diets for early lactation dairy cows from 15.7 to 44.3 g/kg DM can improve energy intake and performance; however, at 44.3 g/kg DM milk fat concentration and ECM decreased. Soybean oil inclusion at 73.4 g/kg DM adversely affected energy intake, fiber digestion and performance of early lactation dairy cows and is not recommended.