Creatinine recovery from bovine urine under the effect of different times and temperatures of storage.

Creatinine is a urinary marker used widely in ruminant's experimental trials. However, despite its great importance no data were found in the literature about the best way to store bovine urine samples. In the sheep urine, was observed an increase in the urinary concentration of creatinine when it was stored acidified (pH 2.5 to 3.5) at a temperature of 28 to 39 °C for 150 days of storage. Nevertheless, urine should be stored acidified (pH below 3) to avoid purine derivative degradation, So, aimed to evaluate creatinine recovery in bovine urine as a function of storage time and temperature. A total of 25 animals' urine (10 Nellore cattle and 15 Holstein cattle) were collected. The urine (40 mL) was diluted in 160 mL of distilled water and its pH was corrected to a value lower than 3 using sulfuric acid drops. A sample of the diluted urine was analyzed to obtain the creatinine concentration reference value on the collection day. The remaining urine was fractionated and preserved at room temperature, cooled (4 °C) or frozen (-20 °C and -40 °C). In the urine of five Holstein cattle was added creatine solutions (20, 40 and 60 mg/dL) to evaluate the creatine to creatinine conservation. These urine samples were analyzed on different days after collection (1, 3, 7, 15, 30 and 45 days). The urine without any added creatine was analyzed on Days 1, 3, 7, 10, 15, 30, 45, 60, 90, 120, and 150 of storage. The addition of creatine in the urine caused an increase in the creatinine concentration (P < 0.05) after 30 days of storage at room temperature and under refrigeration (4 °C). In frozen samples, there was no change in creatinine concentration (P > 0.05). However, creatinine recovery was constant (P > 0.05) until day 15 of storage, regardless of the temperature used, when creatine was not added. After 30 days of storage, an effect of time and/or temperature was observed on creatinine recovery (P < 0.05). Urine samples can be stored at any temperature for up to 15 days after collection to estimate the creatinine concentration. Samples that need storage times longer than 15 days should be frozen (at -20 °C and -40 °C) to avoid creatinine concentration variation.

A Standard Procedure for In Vitro Digestion Using Rumen Fermenters: A Collaborative Study.

Our objective was to propose and evaluate a standard procedure for the evaluation of in vitro dry matter digestibility for ruminant feeds, using artificial fermenters. A collaborative study was performed with seven feed analysis laboratories and four feeds (Tifton 85 hay, corn silage, soybean hulls, and soybean meal). Two types of artificial fermenters were evaluated (DaisyII Ankom and TE-150 Tecnal). Each laboratory received 80 sealed filter bags with samples (20 per feed), eight blank filter bags, a plastic bag with buffer solution reagents, and instructions describing how to conduct a 48 h in vitro assay using an artificial fermenter and how to collect bovine ruminal inoculum. On average, the contribution of laboratory effect to the total random variance was 24%, being less than the contribution of equipment (42%) and error (34%). The repeatability ranged from 3.34 to 5.79%, across feeds. The reproducibility ranged from 5.93 to 8.94% across feeds, which implied Horwitz ratios ranging from 2.94 to 4.10. Due to the specific characteristics of the analytical entity evaluated here, which is defined by the method itself, the proposed method was considered reproducible. The results highlighted that, if the method is followed exactly, its results are precise and present adequate levels of repeatability and reproducibility.

Protein requirements of hair sheep raised in warm areas: a multi-study approach.

The protein requirements of hair sheep are a key factor in increasing profitability and decreasing the environmental impacts of nitrogen excretion. The objective of this study was to evaluate the protein requirements of hair sheep. A database with 382 individual records (269 intact and 113 castrated males) comprising information from 11 studies was used. The studies provided different levels of metabolisable protein intake (MPI) and of N retention, allowing the development of equations to predict the net protein (NP) and the metabolisable protein (MP) requirements. The efficiency of MP use for gain (kpg) was calculated using the equation of daily protein retained against daily MPI above maintenance. The efficiency of MP use for maintenance (kpm) was computed as the ratio between the NP for maintenance (NPm) and the MP for maintenance (MPm). The NPm (1.32 g/kg0.75 EBW) did not differ between sex. The kpm was 0.34, and the kpg was 0.25. The MPm estimated was 3.21 g/kg0.75 BW. Sex affects the protein requirements for gain (NPg). The protein requirements of hair sheep differ from those recommended by feeding systems for sheep. The equations provided herein may improve the optimisation of protein nutrition of sheep, thereby minimising the environmental impacts of sheep production.

Macromineral and trace element requirements for Santa Ines sheep.

Minerals play an important role in animal metabolism. Knowledge of mineral requirements allows well-formulated diets to be provided, which is the main factor that affects performance. To determine the macromineral and trace element requirements for growth and maintenance, thirty-eight 2-month-old Santa Ines lambs with initial body weight (BW) of 13.0 ± 1.49 kg were distributed in a factorial design with feeding levels (ad libitum, 30% and 60% feed restriction) and sex classes [castrated (CM) and intact males (IM)]. The net mineral requirements for gain were higher (P < 0.05) with increasing BW and average daily gain, except for Ca and Na, which remained constant as the empty BW (EBW) increased. The macromineral net requirement for maintenance (g/kg EBW0.75) and the true retention coefficient (k; %) were 0.0784 and 65.2 for Ca, 0.0926 and 80.0 for P, and 0.0379 and 59.0 for K, respectively. The k of Mg was higher (P < 0.05) for IM (11.3 for IM and 7.9 for CM). Sex did not affect (P > 0.05) the maintenance requirement of the trace elements Co, Cu, Zn and Cr which were 0.0015, 0.037, 0.698, and 0.0055 (mg/kg EBW0.75), respectively. Our study indicated that the Santa Ines net mineral requirements are different from the main nutritional requirements established by committees for sheep, which may result in unbalanced diets.

Estimating purine derivatives and nitrogen compound excretion using total urine collection or spot urine samples in grazing heifers.

The study aimed to evaluate the excretion of purine derivatives (PDs) and nitrogen compounds (NCs) and their ratios with creatinine in supplemented Zebu heifers kept on pastures by comparing total urine collection and spot sampling. Five Nelore heifers (400 ± 15 kg) were used in a 5 × 5 Latin square design. The treatments were the amount of concentrate (220 g of crude protein/kg dry matter) offered (0, 3, 6, 9 and 12 g/kg BW). In each period, the total urine collection was performed continuously for 3 days (subsampled at intervals of 4 h, 00:00-04:00 h, 04:00-08:00 h, 08:00-12:00 h, 12:00-16:00 h, 16:00-20:00 h and 20:00-24:00 h). The spot urine samplings were performed (in each period) for 24 h (0, 4, 8, 12, 16 and 20 h). Creatinine, total urinary nitrogen (UN), urea nitrogen (UreaN), allantoin and uric acid were analysed. Creatinine excretion was 23.01 ± 0.19 mg/kg BW and was not affected by collection day, treatment or their interactions (p > 0.05). Treatments affected (p < 0.05) PD excretions, however did not affect the ratio PD:creatinine (p > 0.05). Treatments and collection time affected (p < 0.05) NC excretion, whereas the UN:creatinine and UreaN:creatinine ratios were not affected (p > 0.05). Creatinine excretion and the PD:creatinine ratios in the urine samples estimated by the total or spot sampling were not different (p > 0.05). However, sampling method affected (p < 0.05) the UN:creatinine and UreaN:creatinine (p < 0.05) ratios. Creatinine can adequately estimate urinary excretion in grazing heifers, and a single spot urine sample at any time of the day can be used to estimate PD excretion in grazing heifers. But two spot urine samples are needed for proper NC excretion estimations in grazing heifers' urine.

Effects of exogenous amylase on the in vitro digestion kinetics of whole-crop maize silages made from flint or dent grain type at different phenological stages grown in tropical condition.

The effect of exogenous amylase on the in vitro rumen digestion kinetics of whole-crop maize silage made from dent (RB9004) or flint grain type (RB9308) was evaluated at different phenological stages: soft dough (SOD), early dent (EAD), ½ milkline (½M) and ¾ milkline (¾M). Forage was harvested from 70 to 110 days after sowing. Two rumen-cannulated cows receiving or not exogenous amylase (0.7 g/kg dry matter-DM, provided to achieve 396 kilo Novo units of amylase activity/kg of TMR DM) were used as donor of ruminal fluid. The in vitro gas production kinetics was evaluated according to a dual-pool logistic model. The chemical composition and gas production kinetics were affected by the hybrid and phenological stages. The flint hybrid had lower range for chemical analysis among physiological stages. Harvesting at ½M and ¾M improved DM content, bromatological composition and silage quality parameters compared to dent or flint types. Amylase (i) increased methane (CH4 ) production and in vitro dry matter digestibility (IVDMD) in ½M stage, (ii) improved digestion kinetics by reducing lag time and increasing total gas production and fermentation rates of non-fibrous carbohydrates (NFC) and fibrous carbohydrates (FC), and (iii) increased extent and fermentation rate of NFC and increased fermentation rate of FC fraction in whole-crop maize silages produced from dent or flint types in all phenological stages. Harvesting between ½M and ¾M is the best phenological stage to improve chemical composition and silage quality parameters. Exogenous amylase showed improvements on fibre digestion of silages at ½M and ¾M phenological stages in both grain types of corn.

In vitro screening of plants from the Brazilian Caatinga biome for methanogenic potential in ruminant nutrition.

Thirty-nine plants naturally found in Brazilian Caatinga semiarid biome were screened using an in vitro fermentability testing focused in apparent organic matter digestibility (aOMD), gas, methane (CH4), and short-chain fatty acid (SCFA) production. Three independent in vitro runs were carried out and plants were classified by CH4 concentration as proportion of gas and per unit of apparent digested organic matter (aDOM). According to its CH4 concentration on produced gas (mL/L), the plants were classified as low (> 110), medium (from 60 to 110), and high (< 60) anti-methanogenic potential. From evaluated plants, 3, 24, and 12 were classified as high, medium, and low anti-methanogenic potential. High anti-methanogenic potential plants Cnidoscolus phillacanthus (CnPh), Chloroleucon foliolosum (ChFo), and Anadenanthera macrocarpa (AnMa) produced 21.3, 34.3, and 35.9 mL CH4/L of gas. Methane concentration for Myracrodruon urundeuva (MyUr) was 61.1 mL/L and classified as medium potential. However, CH4 production per unit of aDOM was similar between MyUr and AnMa (3.35 and 2.68 mL/g, respectively). Molar proportions of acetate and propionate in SCFA produced by ChFo fermentation were 0.02 and 0.78 mmol/mol. Acetate to propionate ratios were 0.79, 0.03, 1.39, and 1.36 for CnPh, ChFo, AnMa, and MyUr, respectively. Greater aOMD were observed for Opuntia sp. and Calotropis procera (632 and 601 g/kg, respectively), which were classified as medium mitigating potential plants. AnMa, ChFo, CnPh, and MyUr are promising anti-methanogenic plants for ruminants. Selecting forages to feed ruminants in Caatinga is a potential strategy for enteric CH4 emission reduction, and our in vitro results can support future research by indicating species to be evaluated in in vivo studies integrating mixed diets with performance, digestibility, and CH4 production, yield, and intensity. Graphical abstract ᅟ.

Energy and protein requirements of young Holstein calves in tropical condition.

Our objective was to estimate the energy and protein requirements of Holstein young calves up to 87 days old. Forty-two Holstein calves aged 4 days were used. From these, ten were randomly selected and slaughtered to compose the baseline slaughter (BS) group. The remaining calves were randomly assigned to eight treatments in a 4 × 2 factorial using a completely randomized design. The treatments were 2, 4, 6, and 8 kg/day of milk (297 g/kg CP; 319 g/kg of fat) fed twice daily, and then harvested at 59 or 87 days of age. Calves were fed a starter (193 g/kg CP; 133 g/kg NDF) ad libitum in all treatments. The calves slaughtered at 87 days of age had milk replaced by Coast-cross (Cynodoon sp.) hay (125 g/kg CP; 728 g/kg NDF). The net requirement of energy for maintenance (NEm) was 0.36 MJ/kg EBWG, with efficiency of utilization of energy for maintenance (k m) of 59.4 %. The equation obtained to estimate the net requirement of energy for gain (NEg; MJ/d) was 4.40 × EBW(0.099), with the efficiency of utilization of energy for gain (k g) equal to 46.5 %. The observed requirements of net and metabolizable protein for maintenance (NPm and MPm) were 3.67 and 3.88 g/kg BW(0.75), respectively. The efficiency of use of metabolizable protein for maintenance (k pm ) was 94.6 %. The predicted requirements were higher than those observed in the literature, and this study demonstrated that the requirements of newborn calves are higher than the recommended.

Nutritional and productive performance of dairy cows fed corn silage or sugarcane silage with or without additives.

The objective of this study was to compare the intake, digestibility, and performance of dairy cows fed corn silage, fresh sugarcane, and sugarcane ensiled in three different forms. Twenty-five Holstein cows at 114 ± 12.6 days in milk (DIM) were used. A randomized block design was adopted, using an arrangement of repeated measures over time. The following treatments were tested: corn silage (CS); fresh sugarcane (FS); sugarcane silage without additives (SCS); sugarcane silage enriched with calcium oxide at 5 g/kg of forage (SCSc); and sugarcane silage enriched with Lactobacillus buchneri at 5 × 10(4) cfu/kg of forage (SCSb). The roughage to concentrate ratio was 60:40 for the CS diet and 40:60 for the sugarcane-based diets. The dry matter intake (DMI) as a function of body weight had a downward trend for the cows fed sugarcane silage, compared with those fed FS. The sugarcane silages had higher digestibilities of dry matter (DM), organic matter (OM), and neutral detergent fiber (NDFap), compared with FS. The use of L. buchneri or calcium oxide improved the diet's digestibility. The use of FS, sugarcane silage, or sugarcane silage with additives had no effects on milk and fat-corrected milk yield, compared to corn silage. Cows fed FS presented lower milk total solids content and had a downward trend for milk fat, compared with cows fed sugarcane-silage diets. Cows fed sugarcane silages produced milk with higher casein stability in the alcohol test than cows fed fresh-sugarcane diet. Sugarcane silage, with or without additives, did not reduce the intake of dairy cows, and the use of additives improved the fiber's digestibility.