Effects of supplementation with corn distillers’ dried grains on animal performance,nitrogen balance,and enteric CH4 emissions of young Nellore bulls fed a high-tropical forage diet

The inclusion of corn-dried distillers’ grains (DDG) could be an alternative supplement to increase animal performance, nitrogen efficiency usage (NEU), and decrease enteric methane (CH₄) emissions. Our goal was to determine whether DDG could replace a traditional supplement (cottonseed meal) without affecting animal performance, N balance, and CH₄ emissions. The experiment was conducted during the forage growing season (December to April), with 15 d adaptation, and a 112 d experimental period. The experimental design was completely randomized with four treatments: a mineral supplement (MS), cottonseed meal supplement (CS), 50% replacement of CS by DDG (50DDG), and 100% replacement of CS by DDG (100DDG). Cottonseed meal and DDG were used as protein supplement. A total of 12 paddocks, 3 per treatment, were used to measure forage mass: morphological and chemical composition of forage, forage allowance, and animal performance. Six animals per treatment were used to evaluate DM intake, digestibility, CH₄ emissions, microbial protein production (MCP), and NEU of each treatment. Eighty-one Young Nellore bulls (48 testers, 12 per treatments and 33 adjusters) with initial BW of 255 ± 5 kg (10–12 months old) were supplemented with each supplement type at a level of 0.3% of BW. Pasture management was continuous stocking with a variable stocking rate (put-and-take). Enteric CH₄ was measured using the gas tracer technique. The MCP was quantified using purine derivatives and the NEU mass balance. No differences were found in nutrient intake (P > 0.228). Individual animal performance and gain per area were higher in the treatments with concentrates compared with that of MS; however, there was no difference among treatments CS, 50DDG, and 100DDG. The ADG was 0.83 for MS and 1.08 kg/animal/d when supplemented (P < 0.05). Gain per hectare was 709 kg/ha for MS and 915 kg/ha when supplemented with concentrates (P < 0.05). There was no difference in CH₄ production among treatments that average 180 g/animal/d; however, CH₄ per kg of gain was reduced with CS. The CH₄ conversion factor averaged 5.91%. There was no difference in the synthesis of MCP and NEU. Corn DDG can replace 100% of cottonseed meal as a protein source for supplementation of young Nellore bulls grazing in tropical pastures without affecting animal performance, NEU, MCP, and CH₄ emissions.     

Growth rate,health and welfare in a dairy herd with natural suckling until 6–8 weeks of age: a case report

Over a period of two years, growth rate and health were measured for dairy calves allowed to suckle their mothers up to 6–8 weeks of age. Thirty-one calves were weighted weekly, and the mean daily growth rate was 1.2 ± 0.03 kg from birth up to 13 weeks of age. Illness in calves and young stock was not observed. In the cows, the mean incidences of ketosis, displaced abomasum, puerperal paresis, mastitis, teat injury and retained placenta were 0, 0, 8, 22, 1 and 1%, respectively, during a 6-year period. The mean daily gain of 56 growing bulls was 1.4 kg when slaughtered at 15 months of age, which is higher than the mean daily gain of 0.95 kg in the population. Probiotics, hormones and vaccines were not used, and antibiotics were only used for treating illness. The present study indicates many advantages and few problems when dairy calves are penned together with the cows and allowed natural feeding up to 6–8 weeks of age. This production system was easy to manage, preferred by the farmer, and may satisfy the public concern regarding the practice of immediate separation of cow and calf in commercial milk production.     

The postprandial use of dietary amino acids as an energy substrate is delayed after the deamination process in rats adapted for 2 weeks to a high protein diet

The aim of this study was to determine the contribution of dietary amino acids (AA) to energy metabolism under high protein (HP) diets, using a double tracer method to follow simultaneously the metabolic fate of α-amino groups and carbon skeletons. Sixty-seven male Wistar rats were fed a normal (NP) or HP diet for 14 days. Fifteen of them were equipped with a permanent catheter. On day 15, after fasting overnight, they received a 4-g meal extrinsically labeled with a mixture of 20 U-[¹⁵N]-[¹³C] AA. Energy metabolism, dietary AA deamination and oxidation and their transfer to plasma glucose were measured kinetically for 4 h in the catheterized rats. The transfer of dietary AA to liver glycogen was determined at 4 h. The digestive kinetics of dietary AA, their transfer into liver AA and proteins and the liver glycogen content were measured in the 52 other rats that were killed sequentially hourly over a 4-h period. [¹⁵N] and [¹³C] kinetics in the splanchnic protein pools were perfectly similar. Deamination increased fivefold in HP rats compared to NP rats. In the latter, all deaminated AA were oxidized. In HP rats, the oxidation rate was slower than deamination, so that half of the deaminated AA was non-oxidized within 4 h. Non-oxidized carbon skeletons were poorly sequestrated in glycogen, although there was a significant postprandial production of hepatic glycogen. Our results strongly suggest that excess dietary AA-derived carbon skeletons above the ATP production capacity, are temporarily retained in intermediate metabolic pools until the oxidative capacities of the liver are no longer overwhelmed by an excess of substrates.     

Green tea extracts reduce leukocyte cell–Derived chemotaxin 2 and selenoprotein P levels in the livers of C57BL/6J mice fed a high-fat diet

Epidemiological studies suggest that green tea extracts (GTEs), including catechins such as epigallocatechin gallate and epicatechin gallate, have a beneficial effect on obesity, hyperglycemia, insulin resistance, endothelial dysfunction, and inflammation. Although several studies have shown that catechins directly modulate the cellular and molecular alterations in the liver tissue, the contributions of indirect mechanisms underlying these systemic effects of catechins remain unclear. In this study, we report that, in the C57BL/6J mouse liver, GTEs reduce high-fat diet-induced increases in the levels of hepatokines, liver-derived secretary proteins such as leukocyte cell-derived chemotaxin 2 and selenoprotein P production, which have been shown to induce systemic adverse effects, including several metabolic diseases. These findings suggest that the systemic effects of GTEs involve the regulation of hepatokine production as an indirect mechanism.     

An in silico approach to understand the structure–function properties of a serine protease (Bacifrinase) from Bacillus cereus and experimental evidence to support the interaction of Bacifrinase with fibrinogen and thrombin

Microbial fibrinogenolytic serine proteases find therapeutic applications in the treatment of thrombosis- and hyperfibrinogenemia-associated disorders. However, analysis of structure–function properties of an enzyme is utmost important before its commercial application. In this study, an attempt has been made to understand the structure of a fibrinogenolytic protease enzyme, “Bacifrinase” from Bacillus cereus strain AB01. From the molecular dynamics trajectory analysis, the modelled three-dimensional structure of the protease was found to be stable and the presence of a catalytic triad made up of Asp102, His83 and Ser195 suggests that it is a serine protease. To understand the mechanism of enzyme–substrate and enzyme–inhibitor interactions, the equilibrated protein was docked with human fibrinogen (the physiological substrate of this enzyme), human thrombin and with ten selective protease inhibitors. The Bacifrinase–chymostatin interaction was the strongest among the selected protease inhibitors. The serine protease inhibitor phenyl methane sulphonyl fluoride was found to interact with the Ser134 residue of Bacifrinase. Furthermore, protein–protein docking study revealed the fibrinogenolytic property of Bacifrinase and its interaction with Aα-, Bβ- and Cγ-chains human fibrinogen to a different extent. However, biochemical analysis showed that Bacifrinase did not hydrolyse the γ-chain of fibrinogen. The in silico and spectrofluorometric studies also showed interaction of Bacifrinase with thrombin as well as fibrinogen with a Kd value of 16.5 and .81 nM, respectively. Our findings have shed light on the salient structural features of Bacifrinase and confirm that it is a fibrinogenolytic serine protease.     

SbHKT1;4, a member of the high-affinity potassium transporter gene family from Sorghum bicolor,functions to maintain optimal Na＋/K＋ balance under Na＋ stress

In halophytic plants, the high-affinity potassium transporter HKT gene family can selectively uptake K＋ in the presence of toxic concentrations of Na＋. This has so far not been well examined in glycophytic crops. Here, we report the characterization of SbHKTI;4, a member of the HKT gene family from Sorghum bicolor. Upon Na＋ stress, SbHKT1;4 expression was more strongly upregulated in salt-tolerant sorghum accession, correlating with a better balanced Na＋/ K＋ ratio and enhanced plant growth. Heterogeneous expression analyses in mutants of Saccharomyces cerevisiae and Arabidopsis thaliana indicated that overexpressing SbHKT1;4 resulted in hypersensitivity to Na＋ stress, and such hypersensitivity could be alleviated with the supply of elevated levels of K＋, implicating that SbHKT1;4 may mediate K＋ uptake in the presence of excessive Na＋. Further electrophysiological evidence demonstrated that SbHKT1;4 could transport Na＋ and K＋ when expressed in Xenopus laevis oocytes. The relevance of the finding that SbHKTI;4 functions to maintain optimal Na＋/K＋ balance under Na＋ stress to the breeding of salt-tolerant glycophytic crops is discussed.     

SbHKT1;4, a member of the high‐affinity potassium transporter gene family from Sorghum bicolor,functions to maintain optimal Na~+/K~+ balance under Na~+ stress

In halophytic plants, the high‐affinity potassium transporter HKT gene family can selectively uptake Kt in the presence of toxic concentrations of Nat. This has so far not been well examined in glycophytic crops. Here, we report the characterization of SbHKT1;4, a member of the HKT gene family from Sorghum bicolor. Upon Nat stress, SbHKT1;4 expression was more strongly upregulated in salt‐tolerant sorghum accession, correlating with a better balanced Nat/ Kt ratio and enhanced plant growth. Heterogeneous expression analyses in mutants of Saccharomyces cerevisiae and Arabidopsis thaliana indicated that overexpressing SbHKT1;4 resulted in hypersensitivity to Nat stress, and such hypersensitivity could be alleviated with the supply of elevated levels of Kt, implicating that SbHKT1;4 may mediate Kt uptake in the presence of excessive Nat. Further electrophysiological evidence demonstrated that SbHKT1;4 could transport Nat and Kt when expressed in Xenopus laevis oocytes. The relevance of the finding that SbHKT1;4 functions to maintain optimal Nat/Kt balance under Nat stress to the breeding of salt‐tolerant glycophytic crops is discussed.     

An EPR/HYSCORE, Mössbauer, and resonance Raman study of the hydrogenase maturation enzyme HydF: a model for N-coordination to [4Fe–4S] clusters

The biosynthesis of the organometallic H cluster of [Fe–Fe] hydrogenase requires three accessory proteins, two of which (HydE and HydG) belong to the radical S-adenosylmethionine enzyme superfamily. The third, HydF, is an Fe–S protein with GTPase activity. The [4Fe–4S] cluster of HydF is bound to the polypeptide chain through only the three, conserved, cysteine residues present in the binding sequence motif CysXHisX_(46-53)HisCysXXCys. However, the involvement of the two highly conserved histidines as a fourth ligand for the cluster coordination is controversial. In this study, we set out to characterize further the [4Fe–4S] cluster of HydF using Mössbauer, EPR, hyperfine sublevel correlation (HYSCORE), and resonance Raman spectroscopy in order to investigate the influence of nitrogen ligands on the spectroscopic properties of [4Fe–4S]^2+/+ clusters. Our results show that Mössbauer, resonance Raman, and EPR spectroscopy are not able to readily discriminate between the imidazole-coordinated [4Fe–4S] cluster and the non-imidazole-bound [4Fe–4S] cluster with an exchangeable fourth ligand that is present in wild-type HydF. HYSCORE spectroscopy, on the other hand, detects the presence of an imidazole/histidine ligand on the cluster on the basis of the appearance of a specific spectral pattern in the strongly coupled region, with a coupling constant of approximately 6 MHz. We also discovered that a His-tagged version of HydF, with a hexahistidine tag at the N-terminus, has a [4Fe–4S] cluster coordinated by one histidine from the tag. This observation strongly indicates that care has to be taken in the analysis of data obtained on tagged forms of metalloproteins.     

N2O and CH4 Emissions,and NO3 − Leaching on a Crop-Yield Basis from a Subtropical Rain-fed Wheat–Maize Rotation in Response to Different Types of Nitrogen Fertilizer

Guaranteeing high crop yields while reducing environmental impacts of nitrogen fertilizer use due to associated losses of N₂O emissions and nitrate (NO₃ ^?) leaching is a key challenge in the context of sustainable intensification of crop production. However, few field data sets are available that explore the effect of different forms of N management on yields as well as on N losses in the form of N₂O or NO₃ ^?. Here we report on a large-scale field lysimeter (8 × 4 m²) experiment, which was designed to determine soil CH₄ and N₂O emissions, NO₃ ^? leaching losses and crop yields from a subtropical rain-fed wheat–maize rotation in the Sichuan Basin, one of the most intensively used agricultural regions in China. One control and three different fertilizer treatments with the same total rate of N application (280 kg N ha^?1 y^?1) were included: NF: control (no fertilizer); NPK: synthetic N fertilizer; OMNPK: synthetic N fertilizer plus pig manure; RSDNPK: synthetic N fertilizer plus crop residues. As compared to the standard NPK treatment, annual NO₃ ^? leaching losses for OMNPK and RSDNPK treatments were decreased by 36 and 22%, respectively (P < 0.05). Similarly, crop yield-scaled NO₃ ^? leaching for NPK treatment was higher than those for either OMNPK or RSDNPK treatments (P < 0.05). Direct N₂O emissions for RSDNPK treatment were decreased as compared with NPK and OMNPK treatments (P < 0.05). Furthermore, the yield-scaled GWP (global warming potential) was lower for the treatments where either pig manure or crop residues were incorporated as compared to the standard NPK treatment (P < 0.05). Our study indicates that it is possible to reduce the negative environmental impact of NO₃ ^? leaching and N₂O emissions without compromising crop productivity. Yield-scaled NO₃ ^? leaching, similar to the yield-scaled GWP, represents another valuable-integrated metric to address the dual goals of reducing nitrogen pollution and maintaining crop grain yield for a given agricultural system.     

Low C18 to C20 fatty acid elongase activity and limited conversion of stearidonic acid, 18:4(n–3), to eicosapentaenoic acid, 20:5(n–3), in a cell line from the turbot, Scophthalmus maximus

The TF cell line, derived from a top predatory, carnivorous marine teleost, the turbot (Scophthalmus maximus), is known to have a limited conversion of C₁₈ to C₂₀ polyunsaturated fatty acids (PUFA). To illuminate the underlying processes, we studied the conversions of stearidonic acid, 18:4(n–3), and its elongation product, 20:4(n–3), in TF cells and also in a cell line, AS, derived from Atlantic salmon (Salmo salar), by adding unlabelled (25 μM), U-¹⁴C (1 μM) or deuterated (d5; 25 μM) fatty acids. Stearidonic acid, 18:4(n–3), was metabolised to 20:5(n–3) in both cells lines, but more so in AS than in TF cells. Δ5 desaturation was more active in TF cells than in AS cells, whereas C₁₈ to C₂₀ elongation was much reduced in TF as compared to AS cells. Only small amounts of docosahexaenoic acid (22:6(n–3)) were produced by both cell lines, although there was significant production of 22:5(n–3) in both cultures, especially when 20:4(n–3) was supplemented. We conclude that limited elongation of C₁₈ to C₂₀ fatty acids rather than limited fatty acyl Δ5 desaturation accounts for the limited rate of conversion of 18:3(n–3) to 20:5(n–3) in the turbot cell line, as compared to the Atlantic salmon cell line. The results can account for the known differences in conversions of C₁₈ to C₂₀ PUFA by the turbot and the Atlantic salmon in vivo.