The nutritional significance of endogenous n‐losses along the gastrointestinal tract of farm animals

In animal production, endogenous protein losses associated with the digestion process are important losses, but difficult to measure. Measuring methods include feeding N‐free diets, regression techniques based on amino acid profiles, and separating feed protein and endogenous protein by markers like homoarginine, hydrolysed casein or stable isotopes like ¹⁵N. Endogenous losses arise from saliva, digestive enzymes, bile, shedded epithelial cells and mucins and may be extra stimulated by the presence in feeds of antinutritional factors (ANF) such as lectins, trypsin inhibitors (TI), tannins and fibre. The impact of such factors may differ between non‐ruminants and ruminants. The magnitude of the effect of the different factors is quantified and some of the consequences for protein deposition and nitrogen losses to the environment are discussed.     

Thermal responses to feeding in a secretive and specialized predator (Gila monster, Heloderma suspectum)

We investigate how a unique dietary specialist, the Gila monster (Heloderma suspectum), uses behavioral thermoregulation to elevate body temperature (T_b) after feeding. Lizards in a laboratory thermal gradient were fed rodent meals of three different sizes (5, 10, or 20% of body mass), or sham fed (meal of 0% body mass), and T_bs were recorded for three days before feeding and seven days after feeding. Gila monsters selected a mean T_b of 25.2 °C while fasting (set-point range 23.6–27.1), and increased T_bs after feeding. The magnitude and duration of post-prandial T_b increases are positively related to meal size, and Gila monsters selected mean T_bs up to 3.0 °C higher and maintain elevated T_bs for 3–6 days after feeding. Selection of T_b does not appear to differ between day and night time periods, and because the lizards are both diurnal and nocturnal (at different times of year), photoperiod may not be an important influence on T_b selection.     

锦鲫的摄食代谢与运动代谢及其相互影响

为了探讨锦鲫(Carassius auratus)幼鱼摄食后特殊动力作用(SDA)的变化特征及运动代谢与摄食代谢之间的相互影响,实验首先灌喂锦鲫4%体重的饲料和等体积的纤维素(湿重),测定灌喂前后的耗氧率;另设灌喂饲料、灌喂纤维素、空腹组(对照组)3个组,测定3组的临界游泳速度(Ucrit)和运动耗氧率(MO2);然后在70%、0%临界游泳速度下,分别测定饱足摄食组和空腹组的耗氧率。结果显示:1灌喂饲料后代谢率快速上升,达到峰值后又迅速下降,代谢时间较短,没有一个相对稳定的平台期,灌喂纤维素后代谢率没有显著性变化(P0.05)。提示锦鲫幼鱼的特殊动力作用功率曲线为一个典型的"三角型"模型,且在特殊动力作用总耗能中,生化特殊动力作用占特殊动力作用总耗能的绝大部分,而机械特殊动力作用只占特殊动力作用的极少部分。2锦鲫幼鱼在摄食后临界游泳速度显著下降(P0.05),代谢率显著升高(P0.05)。摄食后的运动过程中,代谢率从摄食开始到代谢率回落至空腹组代谢的标准误范围内的首个数据所对应的时间长度均为6.5 h,且摄食代谢无显著性差异。提示,对锦鲫幼鱼来说,摄食代谢降低了其运动能力,而运动代谢并没有影响摄食代谢。     

Membrane protein shaving with thermolysin can be used to evaluate topology predictors

Topology analysis of membrane proteins can be obtained by enzymatic shaving in combination with MS identification of peptides. Ideally, such analysis could provide quite detailed information about the membrane spanning regions. Here, we examine the ability of some shaving enzymes to provide large‐scale analysis of membrane proteome topologies. To compare different shaving enzymes, we first analyzed the detected peptides from two over‐expressed proteins. Second, we analyzed the peptides from non‐over‐expressed Escherichia coli membrane proteins with known structure to evaluate the shaving methods. Finally, the identified peptides were used to test the accuracy of a number of topology predictors. At the end we suggest that the usage of thermolysin, an enzyme working at the natural pH of the cell for membrane shaving, is superior because: (i) we detect a similar number of peptides and proteins using thermolysin and trypsin; (ii) thermolysin shaving can be run at a natural pH and (iii) the incubation time is quite short. (iv) Fewer detected peptides from thermolysin shaving originate from the transmembrane regions. Using thermolysin shaving we can also provide a clear separation between the best and the less accurate topology predictors, indicating that using data from shaving can provide valuable information when developing new topology predictors.     

Microwave-mediated enzymatic modifications of DNA

Here we report microwave-induced specific cleavage, ligation, dephosphorylation, and phosphorylation of nucleic acids catalyzed by restriction endonucleases, T4 DNA ligase, T4 polynucleotide kinase, and calf intestinal alkaline phosphatase. The microwave-mediated method has dramatically reduced the reaction time to 20 to 50 s. In control experiments, the same reactions failed to give the desired reaction products when carried out in the same time periods but without microwave irradiation. Because the microwave method is rapid, it could be a useful alternative to the time-consuming conventional procedure for enzymatic modification of DNA.     

Peritrophic matrix of Phlebotomus duboscqi and its kinetics during Leishmania major development

Light microscopy of native preparations, histology, and electron microscopy have revealed that Phlebotomus duboscqi belongs to a class of sand fly species with prompt development of the peritrophic matrix (PM). Secretion of electron-lucent fibrils, presumably chitin, starts immediately after the ingestion of a blood meal and, about 6 h later, is followed by secretion of amorphous electron-dense components, presumably proteins and glycoproteins. The PM matures in less than 12 h and consists of a thin laminar outer layer and a thick amorphous inner layer. No differences have been found in the timing of the disintegration of the PM in females infected with Leishmania major. In both groups of females (infected and uninfected), the disintegration of the PM is initiated at the posterior end. Although parasites are present at high densities in the anterior part of the blood meal bolus, they escape from the PM at the posterior end only. These results suggest that L. major chitinase does not have an important role in parasite escape from the PM. Promastigotes remain in the intraperitrophic space until the PM is broken down by sand-fly-derived chitinases and only then migrate anteriorly. Disintegration of the PM occurs simultaneously with the morphological transformation of parasites from procyclic forms to long nectomonads. A novel role is ascribed to the anterior plug, a component of the PM secreted by the thoracic midgut; this plug functions as a temporary barrier to stop the forward migration of nectomonads to the thoracic midgut. This work was supported by the Ministry of Education of the Czech Republic (projects MSM0021620828 and LC06009).     

Effects of oilseed meals and grain–urea supplements fed infrequently on digestion in sheep: 2. Cereal straw diets

An experiment examined the intake, growth responses and rumen digestion of young sheep fed ad libitum oat or barley straws alone or supplemented with approximately isonitrogenous amounts of barley grain and urea (Bar/N), safflower seed meal (SAF) or linseed meal (LIN) supplements provided at 3 day intervals. The supplements comprised 15–22% of total dry matter (DM) intake. Sheep offered either of the straws alone consumed 35.0–37.2 g DM/kg liveweight (LW^0.75) per day of straw and an estimated 2.03–2.07 MJ metabolizable energy (ME) per day, and lost 85–97 g LW per day. Supplements increased (P<0.05 or <0.001) voluntary intake of straw and of total DM, and the organic matter (OM) digestibility of the entire diet. Each of the supplements increased (P<0.001) the estimated ME intake to a similar extent and changed the rapid LW loss of sheep fed straw alone to approximate LW maintenance. Rumen ammonia concentrations in sheep fed barley and oat straws alone (12 and 24 mg NH₃/l, respectively) were expected to be deficient for microbial activity, but were increased (P<0.001) by provision of the supplements. Digestion of straw in synthetic fibre bags incubated in the rumen was markedly increased (P<0.01 or <0.001) when supplements were provided. Rumen pH was depressed briefly to pH <6.0 by the Bar/N, but not by the LIN or SAF, supplements. In young sheep fed cereal straws and losing LW rapidly the oilseed meal supplements increased wool growth more than the barley grain–urea supplements, but both types of supplement increased ME intake similarly and were equally effective to reduce the extent of LW loss.     

Evaluation of Integrated Anaerobic Digestion and Hydrothermal Carbonization for Bioenergy Production

Lignocellulosic biomass is one of the most abundant yet underutilized renewable energy resources. Both anaerobic digestion (AD) and hydrothermal carbonization (HTC) are promising technologies for bioenergy production from biomass in terms of biogas and HTC biochar, respectively. In this study, the combination of AD and HTC is proposed to increase overall bioenergy production. Wheat straw was anaerobically digested in a novel upflow anaerobic solid state reactor (UASS) in both mesophilic (37 °C) and thermophilic (55 °C) conditions. Wet digested from thermophilic AD was hydrothermally carbonized at 230 °C for 6 hr for HTC biochar production. At thermophilic temperature, the UASS system yields an average of 165 L_CH4/kg_VS (VS: volatile solids) and 121 L _CH4/kg_VS at mesophilic AD over the continuous operation of 200 days. Meanwhile, 43.4 g of HTC biochar with 29.6 MJ/kg_{dry_biochar} was obtained from HTC of 1 kg digestate (dry basis) from mesophilic AD. The combination of AD and HTC, in this particular set of experiment yield 13.2 MJ of energy per 1 kg of dry wheat straw, which is at least 20% higher than HTC alone and 60.2% higher than AD only.     

Hydrophilic polydopamine‐coated magnetic graphene nanocomposites for highly efficient tryptic immobilization

In this work, polydopamine‐coated magnetic graphene (MG@PDA) nanocomposites were synthesized by a facile method. Trypsin was then directly immobilized on the surface of the nanocomposites through simple PDA chemistry with no need for introducing any other coupling groups. The as‐made MG@PDA nanocomposites inherit not only the large surface area of graphene which makes them capable of immobilizing high amount of trypsin (up to 0.175 mg/mg), but also the good hydrophilicity of PDA which greatly improves their biocompatibility. Moreover, the strong magnetic responsibility makes them easy to be separated from the digested peptide solution when applying a magnetic field. The feasibility of the trypsin‐immobilized MG@PDA (MG@PDA‐trypsin) nanocomposites for protein digestion was investigated and the results indicated their high digestion efficiency in a short digestion time (10 min). In addition, the reusability and stability of the MG@PDA‐trypsin nanocomposites were also tested in our work. To further confirm the efficiency of MG@PDA‐trypsin nanocomposites for proteome analysis, they were applied to digest proteins extracted from skimmed milk, followed by nano RPLC‐ESI‐MS/MS analysis, and a total of 321 proteins were identified, much more than those obtained by 16‐h in‐solution digestion (264 proteins), indicating the great potential of MG@PDA‐trypsin nanocomposites as the supports for high‐throughput proteome study.     

Variation in faecal digestibility values related to feed efficiency traits of grower-finisher pigs

Providing pigs a diet that matches their nutrient requirements involves optimizing the diet based on the nutrient digestibility values of the considered feed ingredients. Feeding the same quantity of a diet to pigs with similar BW but with different requirements, however, can result in a different average daily gain (ADG) and backfat thickness (BF) between pigs. Digestibility may contribute to this variation in efficiency. We investigated variation in feed efficiency traits in grower-finisher pigs associated with variation in faecal digestibility values, independent of feed intake at the time of measuring faecal digestibility. Considered traits were ADG, average daily feed intake (ADFI), feed conversion ratio (FCR), BF and residual feed intake (RFI). Feed intake, BW, and BF data of one hundred and sixty three-way crossbreed grower-finisher pigs (eighty female and eighty male) were collected during two phases, from day 0 of the experiment (mean BW 23 kg) till day 56 (mean BW 70 kg) and from day 56 to slaughter (mean BW 121 kg). Pigs were either fed a diet based on corn/soybean meal or a more fibrous diet based on wheat/barley/by-products, with titanium dioxide as indigestible marker. Faecal samples of one hundred and five pigs were collected on the day before slaughter and used to determine apparent faecal digestibility of DM, ash, organic matter (OM), CP, crude fat (CFat), crude fibre (CF), and to calculate the digestibility of nonstarch polysaccharides (NSPs) and energy (E). The effects of diet, sex and covariate feed intake at sampling (FIs) on faecal digestibility values were estimated and were significant for all except for CFat. Faecal digestibility values of each individual pig determined at the day before slaughter, corrected for diet, sex and FIs, were used to estimate their association with ADG, ADFI, FCR, BF, and RFI. In the first phase, a one percent unit increase in faecal digestibility of DM, ash, OM, E, CP, CFat, CF, NSP, and Ash individually was related to 0.01–0.03 unit reduction in FCR and 6–23 g/day reduction in RFI. A unit increase in CP digestibility was related to 0.1 mm increase in BF and 10 g/day increase in ADG. In the second phase, a one percent unit increase in faecal digestibility of DM, CP and Ash was related to a decrease of 16–20 g/day in RFI. In conclusion, the relationship between variation in feed efficiency traits and faecal digestibility values is different across the developmental stages of a pig.