Fecal nitrogen to estimate intake and digestibility in grazing ruminants

This study aimed at evaluating forage intake and digestibility in ruminants using fecal nitrogen content, as well as validating a non-linear model to estimate digestibility in ruminants. A total of 58 conventional metabolism trials, carried out with sheep fed 27 forages (offered pure or in mixture) used in Rio Grande do Sul (RS) during the period 1969–1989 was analyzed. OM intake and OM digestibility (OMD) results were regressed linearly against fecal N, and OMD was also estimated from fecal crude protein (N × 6.25) content by a non-linear regression model. Fecal nitrogen excretion estimated forage intake in sheep with an R² = 0.73, whereas a low R² value of 0.36 was observed for OMD estimates. The equation obtained using the non-linear model was OMD = 0.7326 ? 0.3598 exp [(?0.9052 CP (g/kg OM))/100]. The parameters a (0.7326) and b (0.3598) estimated by the equation for all forages were significant (P<0.00001) and there was no effect of type of forage (P=0.38). The mean prediction error (MSPE), was 0.2379, indicating that the equation fit well to the data. The difference between estimated and observed organic matter digestibility was mainly caused by random variation (0.9765). The results indicated that the equation using the non-linear model developed with all forages can be used with enough precision to estimate the OM digestibility of forage consumed by sheep in Rio Grande do Sul.     

Nutrient digestibility in Atlantic salmon and broiler chickens related to viscosity and non-starch polysaccharide content in different soyabean products

Total tract digestibility in Atlantic salmon and ileal digestibility in chicken were assessed from diets with different soyabean products (hulled, toasted, extracted, SBM; reduced oligosaccharide content, ROM; ethanol-extracted protein concentrate, SPC; isolated protein, ISP). The concentration of dietary fibre was highest in SBM and ROM, while it was low in ISP. In vitro viscosity was also higher in SBM than in the other soyabean products. The diets for the salmon and chickens were based on the same feed ingredients, with the exception that fish meal provided half the crude protein in the salmon diets. For each species, the diets were isonitrogenous, contained similar amounts of fat (fish oil), and were balanced with dextrin, thus substituting soyabean non-starch polysaccharides (NSP) and other non-proteinous components by dextrin.In the salmon, total tract digestibility of nitrogen and dry matter were lower (p < 0.05) with the SBM and ROM diets than with the ISP diet. In the chickens, ileal digestibility of dry matter differed among all treatments (p < 0.05). Digestibility of nitrogen, starch, phosphorus and calcium in the chickens was lower (p < 0.05) with the SBM diet than with the ISP and SPC diets, and fat digestibility was lower (p < 0.05) with the SBM and ROM diets than with the SPC diet. Also, the SBM diet gave lower (p < 0.05) digestibility of nitrogen, phosphorus and calcium than the ROM diet, while the ROM diets gave lower (p < 0.05) digestibilities of nitrogen, fat and starch than the ISP and SPC diets. Intestinal viscosity was higher (p < 0.05) and cholesterol content in the blood was lower (p < 0.05) for the chickens fed the SBM-based diet than those fed the other diets. There was a similar response to the different soy products in salmon and chickens with regard to digestibility of nitrogen and fat (p < 0.05). The negative effects of soyabean meals with a low protein content could indicate antinutritive effects of NSP fractions in soyabean meal, as indicated by the elevated viscosity and the decrease in blood cholesterol content.     

Effects of considering the rate of comminution of particles and microbial contamination on accuracy of in situ studies of feed protein degradability in ruminants

Ruminal degradation of dry matter (DM) and crude protein (CP) in samples of soybean meal (SBM), barley grain (BG) and lucerne hay (LH) were measured by an in situ ruminal technique considering either only the outflow rate of particles from the rumen (k_p) or, both k_p and rate of particle comminution (k_c). The effect of correction for the microbial contamination using an ¹⁵N technique was also determined for LH. Degradation and transit studies were completed on three wethers cannulated in the rumen and duodenum and fed a mixed diet of LH and concentrate (2:1 on DM), at an intake level of 50 g DM/kg BW^0.75. Transit studies of concentrates and forages were completed with SBM and LH samples marked with Yb and Eu, respectively. Mean values of k_c were higher in SBM than in LH (0.357 versus 0.204 h, P < 0.05). The same trend was observed for k_p (0.0587 versus 0.0452/h; P < 0.1). Apparent estimates of the proportion of rumen undegraded CP obtained using both rates were 0.884, 0.745 and 0.825 of those obtained using only k_p (0.535, 0.247 and 0.303 for SBM, BG and LH, respectively). Differences occurred for SBM and LH (P < 0.01) and for BG (P < 0.05). The pattern of microbial contamination of LH, expressed as a proportion of residual DM or CP, fitted exponential curves with asymptotic values of 0.076 and 0.586, respectively. Correction for microbial contamination reduced (P < 0.05) estimates of undegraded CP from 0.303 to 0.238, which was further reduced to 0.178 (P < 0.01) when the effect of k_c was also considered.     

Effects of total nonstructural carbohydrates and nitrogen balance on voluntary intake of goats and digestibility of gamagrass hay harvested at sunrise and sunset

We evaluated differences in composition of Iuka gamagrass (Tripsacum dactyloides L.) hay harvested at 06:00 (AM harvest) or 18:00 h (PM harvest), and measured how protein supplementation and time of harvest interact to affect the voluntary intake, digestibility, and N balance of goats. Boer cross wethers (n = 28; 24 kg) were randomly assigned to be fed supplement (310 g/kg of crude protein (CP), fed at 110 g/kg of dry matter (DM) intake, 14 goats) or no supplement (14 goats). Within supplemented or not supplemented groups, goats were randomly assigned to a crossover design of AM harvest (seven goats) or PM harvest (seven goats), and housed individually in metabolism crates with free access to water and mineral blocks. They were fed twice daily, with supplement offered 30 min prior to the morning feedings. After a 7-d adaptation, voluntary intake (goats were offered 1100 g/kg of previous day's intake) was measured for 14 d, followed by a 4-d adjustment phase to equalize DM offered between periods, and finally a 5-d digestion and balance phase. After Period 1, goats were switched to their new hay harvest times, and the protocol was repeated. Compared to the AM harvest, the PM harvest had higher (P<0.03) proportions of total nonstructural carbohydrates (TNC, 70.8 g/kg DM versus 59.0 g/kg DM), monosaccharides (37.0 g/kg DM versus 28.6 g/kg DM), di- and polysaccharides (18.5 g/kg DM versus 15.4 g/kg DM) and less neutral detergent fiber (NDF, 700 g/kg versus 710 g/kg). Crude protein (79 g/kg DM) and starch (15.2 g/kg DM) were similar for the PM and AM harvest. Dry matter digestibility was higher (P<0.03) for the PM versus AM harvests (555 g/kg DM versus 531 g/kg DM) and for supplemented versus not supplemented (563 g/kg DM versus 522 g/kg DM). Voluntary gamagrass DM intake (550 g/d versus 548 g/d) and calculated total digestible DM intake (327 g/d versus 313 g/d) were similar for the PM and AM harvest. However, total digestible DM intake during the digestion and balance phase was higher (P<0.01) for the PM versus AM harvest (317 g/d versus 299 g/d). Time of harvest did not affect N intake, digestion, or calculated retention. Compared to no supplementation, the supplement improved (P<0.01) N digestion (6.1 g/d versus 3.7 g/d) and retention (2.2 g/d versus 1.1 g/d). The PM harvest increased DM digested, largely TNC and digestible DM intake by goats due to increased TNC and not because of a 2% increase in DM intake. Providing a protein supplement had very limited effects on intake and digestibility of gamagrass.     

In vitro estimations of the rate and extent of ruminal digestion of starch-rich feed fractions compared to in vivo data

An experiment was conducted to study the rumen digestion characteristics of whole feeds (WF) and the neutral detergent fibre (aNDF) and neutral detergent soluble (NDS) fractions of a range of starch-rich feeds using an automated in vitro gas production (GP) technique. In addition, the ruminal digestibility values predicted from the GP data were compared to previously acquired in vivo data. Nine feeds with starch concentrations ranging from 389 to 712 g/kg dry matter and with known in vivo digestibilities were subjected to neutral detergent extraction. The GP for each WF and the corresponding aNDF fractions were measured in duplicate in buffered rumen fluid during 72 h on two occasions. The fermentation residues were collected and analyzed for aNDF concentration to estimate their true organic matter (OM) and NDF digestibility. The GP from the NDS fraction was calculated by subtracting the GP from the aNDF fraction from the GP of the WF. A three-pool Gompertz model was fitted to the GP profiles (R² = 0.99) and a two compartment, mechanistic and dynamic rumen model was used to predict the digestibility of the potentially digestible feed fraction and the effective digestion rate (k_d). The true OM and NDF digestibility determined for the WF ranged from 0.804 to 1.011 and from 0.362 to 1.107, respectively. The NDF digestibility determined for the aNDF fraction ranged from 0.410 to 0.985. The effective k_d values estimated using GP data varied from 0.118 to 0.282/h for the WF and from 0.123 to 0.301/h for the NDS fraction, and were less (P<0.05) for maize compared to small grains (SG) but did not differ between barley and wheat (P>0.05). The effective k_d values for the aNDF fraction ranged from 0.039 to 0.082/h and did not differ (P>0.05) either between maize and SG or between barley and wheat. The predicted ruminal NDS digestibility determined using GP data closely matched the in vivo data describing starch digestion (R² = 0.81). The effective k_d values for the WF were strongly related (R² = 0.94) to those for the NDS fractions. The results indicate that when measured with the GP technique, the differences in the digestion characteristics of maize and small grains are less than those previously reported in studies using the in situ method. It is concluded that the predicted NDS digestibility determined using GP data corresponded well to the in vivo starch digestibility. Our results also suggest that the first order digestion rates of NDS (starch) in starch-rich feeds can be accurately determined by incubating WF samples in the GP system and using the GP kinetic data in a dynamic, mechanistic rumen model.     

In vitro evaluation of starch degradation from feeds with or without various heat treatments

An in vitro method was used to evaluate starch degradation from various feeds with or without heat treatments in four studies. The method was based on incubation of feed samples with a buffered rumen fluid solution and subsequent enzymatic analysis of the remaining starch. In all studies, heat treatment of the feed samples increased rate or extent of starch degradation to glucose. In Study 1, measurements of remaining starch, after 5 h in vitro incubations, demonstrated substantial effects of cooking on starch degradation in potatoes, and a trend to faster degradation from autoclaving peas. Up to 0.60 of the starch remaining after a 5 h of incubation was not recovered by centrifugation at 3000 × g for 10 min. In Study 2, cooking increased in vitro starch degradation rate from isolated potato starch (from 0.038 to 0.197/h). Intact starch in barley and wheat grain had similar rates of degradation (0.117 and 0.109/h, respectively). In Study 3, both autoclaving time (15, 30, 60 min) and temperature (115, 130 and 145°C) affected in vitro starch degradation rates in peas, and, in no case did autoclaving for only 15 min increase degradation rates. For the 30 min autoclaving time, only the highest temperature (145°C) increased the degradation rate of the pea starch compared to the untreated peas (0.175 versus 0.110/h). When autoclaving for 60 min, both 130 and 145°C resulted in a considerable increase in starch degradation rate (0.211 and 0.193/h, compared to 0.110/h for the untreated peas). In Study 4, the proportion of starch degraded at 8 h of in vitro incubation was increased by heat treatment of pure potato starch (0.155 versus 0.870), peas (0.491 versus 0.815), barley (0.686 versus 0.913) and maize (0.351 versus 0.498). Measurements of volatile fatty acid production in the fermentation tubes showed a lower acetate:propionate ratio for the faster fermenting heat-treated feeds. Heat treatment generally increased starch degradation in vitro.     

Effects of non-enzymatic browning reaction intensity on in vitro ruminal protein degradation and intestinal protein digestion of soybean and cottonseed meals

The effects of non-enzymatic browning reactions on in vitro ruminal gas production and in vitro ruminal and intestinal crude protein (CP) digestibilities of soybean (SBM) and cottonseed (CSM) meals were investigated. Non-enzymatically browned SBM and CSM samples were prepared using two xylose levels (10 or 30 g/kg dry matter), two heating lengths (30 or 60 min) and two heating temperatures (120 or 150 °C) for a total of one untreated (commercially solvent-extracted, Control) and eight treated samples for each protein source. The control SBM had higher (P<0.001) in vitro ruminal CP degradability values than the treated samples. Intestinal protein digestibility and total-tract CP digestibility of CSM and SBM were affected by the treatment (P<0.01). The results of the study indicate that not only ruminal CP degradability is reduced but also intestinal and total-tract CP digestibilities may be lowered depending on protein source and intensity of the non-enzymatic browning reaction.     

Effects of exogenous enzymes on in vitro gas production kinetics and ruminal fermentation of four fibrous feeds

This study was conducted to investigate effects of increasing doses: 0 (control), 6 (low), 12 (medium) and 24 (high) mg/g DM of ZADO^® enzyme preparation mixture (ENZ) on in vitro gas production (GP) and some ruminal fermentation parameters of the fibrous feeds Saccharum officinarum (leaves), Andropogon gayanus (leaves), Pennisetum purpureum (leaves) and Sorghum vulgare (straw). Rumen liquor was obtained from two Brown Swiss cows fitted with permanent rumen cannulae fed a total mixed ration of a 500:500 commercial concentrate and alfalfa hay ad libitum. The GP was recorded at 2, 4, 6, 8, 10, 12, 24, 48, 72 and 96 h of incubation. After 96 h, the incubation was stopped and the pH of the mixture was determined and filtrate used to determine dry matter degradability (DMD), partitioning factor (PF₉₆), gas yield (GY₂₄), in vitro organic matter digestibility (OMD), metabolizable energy (ME), short chain fatty acids (SCFA), and microbial crude protien production (MCP). In general, the crude protein (CP) content of the fibrous feeds was low and ranged from 23 g/kg DM (S. officinarum) to 44 (A. gayanus). The fibre contents (i.e., NDFom and ADFom) were highest (P<0.05) in S. officinarum. Increasing ENZ dose linearly increased (P<0.05) GP of all fibrous feeds and had a quadratically increased (P<0.05) asymptotic gas production in P. purpureum and S. vulgare and rate of gas production in S. officinarum and S. vulgare. Addition of ENZ also quadratically increased (P<0.05) GP at all incubation times in S. officinarum and S. vulgare, and A. gayanus, but only at 72 h in A. gayanus. The parameters of ruminal fermentation of OMD, ME, GY₂₄ and SCFA linearly increased (P<0.05) and MCP linearly decreased (P<0.05) with the ENZ addition. Addition of enzyme affected ruminal fermentation of our feeds differently, mainly dependent on their fibre content, although dosage of enzyme was also important as impacts generally increased at higher dosages of ENZ.     

Nutritional characterization of Mucuna pruriens: 3. Effect of replacing soybean meal with Mucuna on intake,digestibility, N balance and microbial protein synthesis in sheep

Mucuna pruriens seeds have relatively high crude protein (CP) concentrations, but little is known about their potential to replace commonly used CP supplements in ruminant rations. The aim of this experiment was to determine effects of replacing soybean meal (SB) with Mucuna on the performance of lambs. Forty Rambouillet lambs (33.2 ± 5.73 kg) fed a basal diet of maize grain, cottonseed hulls and urea were randomly assigned to one of four supplements formulated by substituting 0 (SB), 330 (Lo), 670 (Med) or 1000 g/kg (Hi) of soybean meal with rolled Mucuna seeds. Lambs were housed individually in metabolic crates and allowed ad libitum access to isocaloric (metabolizable energy=11.7 MJ/kg dry matter, DM) and isonitrogenous (CP = 146 g/kg, DM) diets for 14 d of adaptation and 7 d of total fecal collection. Fecal egg counts and coccidian oocyst scores were determined on d 14. Dry matter intake (1.7 kg/d versus 1.5 kg/d; P<0.05), CP digestibility (774 g/kg versus 714 g/kg DM; P<0.05) and N retention (28.0 g/d versus 20.4 g/d; P<0.01) were higher and amylase-pretreated neutral detergent fiber digestibility (617 g/kg versus 686 g/kg DM) was lower (P<0.05) in sheep fed SB versus Mucuna diets. However, supplementary protein source did not affect rumen pH, blood urea N or glucose concentration, or fecal egg counts. Increasing the level of Mucuna supplementation increased (P<0.05) level and efficiency of microbial protein synthesis, ruminal fluid acidity, total volatile fatty acid concentration, decreased (P<0.05) coccidian oocyst scores, and tended (P<0.10) to increase N retention. Therefore, SB is a better supplement than Mucuna to support performance of lambs. Nevertheless, Mucuna seeds are a promising CP supplement for situations where cost or availability precludes use of SB in ruminant rations.     

Effect of 1,25 dihydroxyvitamin D3 on intracellular IFN-γ and TNF-α positive T cell subsets in pulmonary tuberculosis

We studied the immunomodulatory effect of 1,25(OH)₂D₃ on single cell expression of IFN-γ and TNF-α cytokines in T cell subsets of pulmonary tuberculosis (PTB) patients (n = 22) and normal healthy subjects (n = 22). Peripheral blood mononuclear cells (PBMCs) were cultured with live Mycobacterium tuberculosis (MTB) with or without 1,25(OH)₂D₃ (10^?7 M) for 48 h. T cell subsets positive for IFN-γ and TNF-α were enumerated by flow cytometry and the culture supernatants were assayed for both the cytokines using ELISA. In both NHS and PTB patients, a significantly reduced percentage of IFN-γ and TNF-α expressing CD3+, CD3+CD4+ and CD3+CD8+ T cells were observed in cultures stimulated with live MTB and treated with 1,25(OH)₂D₃ compared to cultures without 1,25(OH)₂D₃ (NHS; CD3+ IFN-γ+: p < 0.0001; CD3+TNF-α +: p = 0.0292 and PTB; CD3+ IFN-γ+: p = 0.0292; CD3+ TNF-α +: p = 0.0028). The levels of IFN-γ and TNF-α in the culture supernatants of 1,25(OH)₂D₃ treated cultures were also found to be significantly decreased in both groups (NHS; IFN-γ: p = 0.0001; TNF-α: p < 0.0001) and (PTB; IFN-γ: p < 0.0001; TNF-α: p < 0.0001). A positive correlation was observed between IFN-γ and TNF-α expressing CD3+CD8+ T cells in MTB stimulated cultures treated with or without 1,25(OH)₂D₃ in NHS (p = 0.0001; p = 0.001, respectively) and PTB patients (p = 0.002; p = 0.005, respectively). The present study revealed the suppressive effect of 1,25(OH)₂D₃ on single cell expression of IFN-γ and TNF-α by CD3+CD4+ and CD3+CD8+ T cells in pulmonary tuberculosis. This suppressive effect of 1,25(OH)₂D₃ on proinflammatory and Th1 cytokine positive cells might have a role in reducing inflammation at the site of infection.     

Supplemental vitamin D increases serum cytokines in those with initially low 25-hydroxyvitamin D: A randomized,double blind,placebo-controlled study

The purpose of this study was to determine if vitamin D status before supplementation influences the cytokine response after supplemental vitamin D. Forty-six reportedly healthy adults (mean(SD); age, 32(7) y; body mass index (BMI), 25.3(4.5) kg/m²; serum 25-hydroxyvitamin D (25(OH)D), 34.8(12.2) ng/mL) were randomly assigned (double blind) to one of three groups: (1) placebo (n = 15), or supplemental vitamin D (cholecalciferol) at (2) 4000 (n = 14) or (3) 8000 IU (n = 17). Supplements were taken daily for 35 days. Fasting blood samples were obtained before (Baseline, Bsl) and 35-days after (35-d) supplementation. Serum 25(OH)D, 1,25-dihydroxyvitamin D (1,25(OH)D), cytokines, and intact parathyroid hormone with calcium were measured in each blood sample. Supplemental vitamin D increased serum 25(OH)D (4000 IU, ≈29%; 8000 IU, ≈57%) and 1,25(OH)D (4000 IU, ≈12%; 8000 IU, ≈38%) without altering intact parathyroid hormone or calcium. The vitamin D metabolite increases in the supplemental vitamin D groups (n = 31) were dependent on initial levels as serum 25(OH)D (r = −0.63, p < 0.05) and 1,25(OH)D (r = −0.45, p < 0.05) at Bsl correlated with their increases after supplementation. Supplemental vitamin D increased interferon (IFN)-γ and interleukin (IL)-10 in subjects that were vitamin D insufficient (serum 25(OH)D < 29 ng/mL) compared to sufficient (serum 25(OH)D ⩾ 30 ng/mL) at Bsl. We conclude that supplemental vitamin D increase a pro- and anti-inflammatory cytokine in those with initially low serum 25(OH)D.     

Assessment of the efficacy of varying experimental exogenous fibrolytic enzymes using in vitro fermentation characteristics

This study evaluated a series of recombinant, single activity experimental enzyme products including 13 endoglucanases (END) and 10 xylanases (XY), for their potential to improve in vitro ruminal degradation of alfalfa hay in two experiments. Based on the endoglucanase or xylanase enzymatic activities measured using complex substrates at the optimal conditions (pH 5.4, 37 °C) for the enzymes, a dose level (1 unit/g dry matter [DM]) was chosen for addition of enzymes to substrate. Enzyme products, re-suspended with water, were added to alfalfa hay (0.5 or 1.0 g DM) in culture vials in six replications. Anaerobic buffer medium (20 or 40 ml) adjusted to pH 6.0 and strained ruminal fluid (5 or 10 ml) were sequentially added to the vials and incubated for 18 h. Headspace gas production (GP) was measured throughout the incubation, and degradability of organic matter (OMD) and fibre and volatile fatty acid (VFA) concentrations were determined after 18 h of incubation. The enzyme products had a wide range of added endoglucanase or xylanase activities when determined using pure substrates and physiological conditions typical of the rumen (pH 6.0, 39 °C). In experiment 1, many END, and some XY, products increased GP and OMD. The correlation between added endoglucanase activity determined at ruminal conditions and OMD improvement was high (r = 0.71; P<0.01), whereas added activity of xylanase was not associated with OMD improvement. Two END and two XY products selected from experiment 1 were further assessed because they substantially improved GP and OMD. In experiment 2, all enzyme treatments, alone or in combination, increased total GP and DM and fibre degradabilities (P<0.05). However, the combinations of END and XY did not increase degradation of alfalfa beyond that of the component enzymes. Total VFA production was not affected by enzyme treatments although some products changed the acetate to propionate ratio. Experimental exogenous enzyme products with either endoglucanase or xylanase activity substantially improved in vitro ruminal degradation of alfalfa hay, but further improvement by combining these activities did not occur.     

Losses of particulate N during filtration and handling of feed and rumen incubation residues

Unintended losses of feed N occur during filtration procedures in the laboratory as well as during washing of in sacco residues and other procedures to remove microbial contamination. Our aim was to find the most reliable method for recovery of buffer and neutral detergent (ND) insoluble N (BIN and NDIN, respectively) in feed residues. In Experiment 1, paper filtration (PF; 20–25 μm retention) and sintered glass crucible filtration (GF; 40–90 μm retention) and centrifugation at 3000×g (LSC) were compared for recovery and loss of feed N after feeds were incubated with a borate–phosphate buffer or ND. The same ND removal techniques and centrifugation at 5000×g (HSC) were also applied to in vitro-fermented residues. In Experiment 2a, in vitro fermentation residues were transferred to in sacco bags (28 μm pore openings), washed, extracted with ND using GF or LSC techniques to remove detergents. In 2b, the in vitro residues were homogenised, stomached and/or centrifuged in different ways, then extracted with ND using HSC to remove ND. The NDIN was assumed to exclude microbial N. Samples used in Experiments 1 and 2a consisted of two grasses, birdsfoot trefoil (Lotus corniculatus), wheat bran, rapeseed cake (RSC), dried distillers grain with solubles from wheat (DDGS), de-hulled barley grain (barley) and solvent extracted soybean meal (SBM). For Experiment 2b, we used only one grass and barley. Highest recovery of BIN was by PF > GF > LSC, whereas NDIN recovery was highest for LSC with no differences between filtration techniques for most feeds. After in vitro fermentation, recovery of NDIN was 1.2–7.1- and 1.2–2.8-fold higher by LSC versus GF and PF, respectively. Washing of in vitro residues in sacco caused NDIN reductions of up to 0.35 and 0.85 when GF or LSC was used to remove ND. Losses of NDIN from in vitro fermented feeds, including in sacco and GF losses after ND extraction, were 0.64–0.70 in forage, and 0.81 and 0.83 in SBM and DDGS, respectively. Losses of NDIN from homogenisation, pummelling and centrifugation of in vitro residue were smaller (0.02–0.42) than after sample washing in sacco bags. Oven drying of in vitro residues at 65 °C prior to ND extraction inflated NDIN values and must be avoided. A higher centrifugal force, i.e., 5000×g instead of 3000×g and fewer centrifugations (three instead of five) resulted in the highest recovery and are recommended to minimise sample losses during centrifugation of in vitro residues.     

Intake,digestibility and nitrogen utilization of Robinia pseudoacacia foliage fed to growing goat wethers

Black locust (BL; Robinia pseudoacacia L.), a native tree of southeastern USA known to contain substantial levels of condensed tannins (CT), was fed to 32, 4 month old (20.4 kg BW) Boer cross wether goats in two randomized complete block design trials. The objectives were to examine the effects of feeding hay diets containing several levels of fresh BL foliage on intake, digestibility, and N metabolism. First year (1999) diets were H_E (100% Eastern gamagrass [EGH; Tripsacum dactyloides L.] hay), H_EG (70% EGH and a 30% mixture of 59% ground corn [GC; Zea mays L.], 36% soybean meal [SBM; Glycine max L.], and 5% minerals), 25BL99 (75% EGH and 25% BL leaves), and 50BL99 (50% EGH and 50% BL leaves). Second year (2000) diets were H_O (100% orchardgrass [OGH; Dactylis glomerata L.] hay), H_OG (70% OGH and a 30% mixture of 63% GC, 37% SBM, and 5% minerals), 50BL00 (50% OGH and 50% BL leaves), and 75BL00 (25% OGH and 75% BL leaves). In 1999, apparent digestibilities of the diets in the order listed above were 62.4, 68.2, 58.0, and 60.6% (P = 0.001) for DM and 62.8, 72.5, 56.0, and 59.1% (P = 0.001) for crude protein (CP). Acid detergent lignin digestibilities were negative for diets 25BL99 (−56.7%) and 50BL99 (−49.3%), apparently due to the formation of insoluble tannin and lignin complexes during passage through the digestive tract. Intakes of DM were similar across diets. In 2000, apparent digestibilities of diet DM (64.4, 71.7, 64.8 and 65.4%) and CP (70.0, 76.0, 66.6, and 66.5%) did not differ. Lignin digestibilities were positive for diets 50BL00 (9.4%) and 75BL00 (29.6%) unlike those for year 1999. Overall, BL contained 10% CT and 18–34% hydrolyzable tannins. In 1999, N intake, urinary N (UN) excretion and N retained were higher for diet H_EG (P = 0.01) than diet H_E whereas fecal N excretion (FN) was similar for diets H_EG, 25BL99 and 50BL99. In 1999, FN excretion as a percentage of N intake was higher (P < 0.02) in the BL diets, although UN as a percentage of N intake did not differ among diets. In 2000, N intake and FN output were higher for BL diets compared to diets H_O (P = 0.01) and H_OG (P = 0.02). Fecal N as a percentage of N intake was lower (P = 0.01) for diet H_OG (24.0%) than for diets 50BL00 (33.4%) and 75BL00 (33.5%). Conversely, urinary N as a percentage of N intake was higher for diets H_O and H_OG compared to the BL diets (P = 0.02). Increased levels of dietary BL increased FN, suggesting that tannins formed insoluble protein complexes thus hindering digestibility.     

Nitrogen in vivo digestibility and in situ degradability data for estimation of lower tract N digestibility with or without correction for microbial contamination

In situ degradability and in vivo (by difference) digestibility trials were conducted to estimate lower tract residual N digestibility (LTRND) of five protein supplements. Efforts were also made to improve the in situ method of measuring protein degradability. For in situ degradability trials, soybean meal (SBM), corn gluten meal (CGM), cotton seed cake (CSC), wheat bran (WB) and corn gluten feed (CGF) were weighed into Dacron bags and incubated in the rumen of three cannulated Chios ewes. SBM, CGF and WB were degraded significantly, while CGM and CSC were least degraded. Microbial contamination (MC) resulted in a 5.3–28.3% artificially decrease in effective ruminal protein degradation of supplements. Total tract digestibility was measured using five rams in an in vivo, by difference, trial using a 5 × 5 Latin-square design. SBM had higher CP digestibility compared to WB, CGF and CSC, and higher N free extract (NFE) digestibility compared to the other feeds. CGM showed higher CP digestibility compared to WB, CGF or CSC, while CGF had higher organic matter (OM) and crude fibre (CF) digestibility compared to WB. CSC was the protein source with the lowest digestibility of OM, CP and NFE in comparison with the other feeds. LTRND was predicted as 0.928, 0.806, 0.227, 0.540, and 0.498 for SBM, CGM, CSC, WB, and CGF, respectively, or 0.931, 0.803, 0.147, 0.364, and 0.316 when the correction for MC was applied. Lower tract N digestibility could be predicted via a combination of in situ degradability and in vivo apparent digestibility data. This approach yields significant data regarding LTRND estimation of protein supplements, while diminishing animal suffering by avoiding small intestinal fistulation.     

Forming nanoparticles of α-amylase and embedding them into solid surfaces

In the current work nanoparticles (NPs) of α-amylase were generated in an aqueous solution using high-intensity ultrasound, and were subsequently immobilized on polyethylene (PE) films, or polycarbonate (PC) plates, or on microscope glass slides. The α-amylase NPs coated on the solid surfaces have been characterized by ESEM, TEM, FTIR, XPS and AFM. The substrates immobilized with α-amylase were used for hydrolyzing soluble potato starch to maltose. The amount of enzyme introduced in the substrates, leaching properties, and the catalytic activity of the immobilized enzyme were compared. The catalytic activity of the amylase deposited on the three solid surfaces was compared to that of the same amount of free enzyme at different pHs and temperatures. α-Amylase coated on PE showed the best catalytic activity in all the examined parameters when compared to native amylase, especially at high temperatures. When immobilized on glass, α-amylase showed better activity than the native enzyme over all pH and temperature values studied. However, the immobilization on PC did not improve the enzyme activity at any pH and any temperature compared to the free amylase. The kinetic parameters, K_m and V_max were also calculated. The amylase coated PE showed the most favorable kinetic parameters (K_m = 5 g L⁻¹ and V_max = 5E−07 mol mL⁻¹ min⁻¹). In contrast, the anchored enzyme-PC exhibited unfavorable kinetic parameters (K_m = 16 g L⁻¹, V_max = 4.2E−07 mol mL⁻¹ min⁻¹). The corresponding values for amylase-glass were K_m = 7 g L⁻¹, V_max = 1.8E−07 mol mL⁻¹ min⁻¹, relative to those obtained for the free enzyme (K_m = 6.6 g L⁻¹, V_max = 3.3E−07 mol mL⁻¹ min⁻¹).     

Intake,digestion and selection of roughage with different staple lengths by sheep and goats

The diet selection, ingestive and digestive responses of goats and sheep offered a single forage, which was prepared with three staple lengths: long (L), medium (M) and short (S) (mean particle size (mm): 13.29, 7.26 and 0.69, respectively) were studied in three experiments. The sheep (Scottish Blackface wethers) increased their dry matter intake (sheep DMI: 60.1, 61.1 and 66.2 g DM/(kg W^0.75) per day for L, M and S) and reduced mean retention time (MRT) of the undigested residues (sheep MRT: 54.6, 52.9 and 45.9 h for L, M and S) and digestive efficiency (sheep DM digestibility: 0.524, 0.522 and 0.493 for L, M and S) with the reduction in forage particle size. The respective responses of goats (feral crosses) were not modified with forage staple length (goat DMI: 68.1, 65.9 and 67.3 g DM/(kg W^0.75) per day for L, M and S); (goat MRT: 38.7, 39.3 and 41.1 h for L, M and S); (goat DM digestibility: 0.495, 0.475 and 0.480 for L, M and S). However, both species had similar intakes of digestible dry matter (DDMI) on all staple lengths (mean DDMI: 32.0 and 32.6 g DDM/(kg W^0.75) per day for sheep and goats). On the longer staple lengths (L and M) goats masticated the fibre into smaller particles than did sheep and had (P<0.01) a greater proportion of small particles in their boli (mean: 0.45 and 0.30 for goats and sheep). The selection of components within the forage offered was different for the two species. Goats consumed forage of lower nitrogen (N) (P<0.01) and in vitro organic matter digestibility (OMD) (P<0.05) and higher neutral detergent fibre (NDF) (P<0.01) contents than sheep. This was associated with the residues from sheep having a coarser texture than those of goats. When the forages of different staple lengths were offered in pairs to the animals, there was no evidence for selection of forage types by either species. It would appear that the greater ability of goats to chew efficiently provides a reason for the different responses to staple length in intake, digestibility and MRT by the two species.     

Nutritional characterization of Mucuna pruriens: 4. Does replacing soybean meal with Mucuna pruriens in lamb diets affect ruminal,blood and tissue l-dopa concentrations?

The harmful effects of the 3,4-dihydroxy-l-phenylalanine (l-dopa) in Mucuna pruriens (Velvet bean) seeds have limited its use as a protein supplement for monogastrics and humans. Little is known about the extent of metabolism of Mucuna l-dopa in ruminants or its accumulation in ruminant tissues consumed as food by humans. This study aimed to determine if replacing soybean meal (SB) with Mucuna increases concentrations of l-dopa in rumen fluid, blood and muscle tissue of lambs. Twenty-seven RM lambs (RM; initial body weight, BW = 33.8 ± 5.44 kg) and 12 Florida Native (FN; initial BW = 24.9 ± 8.63 kg) lambs were assigned to four treatments and fed a basal diet of coastal bermudagrass hay, corn grain, and liquid molasses for 42 (FN) or 49 days (RM) and then slaughtered. Dietary supplements were formulated by substituting 0 (SB), 330 (Lo), 670 (Med) or at least 1000 (Hi) g/kg of SB with rolled Mucuna seeds (l-dopa, 24 g/kg dry matter, DM). Body weight was measured weekly and carcass characteristics and concentrations of l-dopa in rumen fluid, blood and the sterno-mandibularis muscle were measured at slaughter when concentrations of blood urea N, glucose, haptoglobin and cerruloplasmin were also measured. Lambs fed SB had higher (P<0.05) average daily gain than those fed Mucuna (0.20 versus 0.15 kg/day for RM; 0.21 versus 0.14 kg/day for FN). However, concentrate protein source did not affect dressing and concentrations of blood urea N, or blood glucose. Feeding the Hi diet versus the SB diet did not increase concentrations of blood cerruloplasmin, or l-dopa or concentrations of l-dopa metabolites in blood. No l-dopa was found in the ruminal fluid of the lambs and l-dopa concentrations in the sterno-mandibularis muscle were low (i.e., <5 ng l-dopa/g), and unaffected (P>0.05) by diet. Ingested Mucuna l-dopa was extensively metabolized in lambs, and did not accumulate to toxic levels in muscle tissues.     

Clinical profile of sheep fed non-conventional feeds containing phenols and condensed tannins

A study was carried out to investigate the effects of feeding low quality non-conventional feeds (NCF) containing phenols and condensed tannins on the clinical profiles of sheep. Thirty-two Omani sheep were fed one of four diets with two base roughages, urea-treated palm frond (UTPF) and Rhodesgrass hay (RGH) and two concentrates, commercial concentrate (CC) and a by-products concentrate (BC) for 120 days. Haematological, serum biochemical and urine analyses were used to assess sheep health. Non-conventional feeds (urea-treated palm frond and by-products concentrate) contained higher levels of polyphenols and condensed tannins than conventional feeds (Rhodesgrass hay and commercial cubes). Feeds based on urea-treated palm frond had lower dry matter, crude protein, acid detergent fibre, neutral detergent fibre, gross energy (P < 0.001) and ash (P < 0.05) digestibility coefficients than those based on Rhodesgrass hay. Animals fed NCF had lower feed intake (P < 0.001) and lower body gain (P < 0.001) than those fed conventional ones. They also produced larger volumes of faeces (P < 0.01) which contained higher levels of nitrogen (P < 0.001) and had lower viscosity values of intestinal content (P < 0.001). Rumen liquor of NCF-fed animals had higher pH and lower ammonia–nitrogen levels (P < 0.01). Animals fed urea-treated palm frond plus by-products concentrate had lower lymphocyte (P < 0.01), monocyte (P < 0.05) and eosinophil (P < 0.05) counts by the end of the trial than those fed Rhodesgrass hay based diets. The urea-treated palm frond and by-products concentrate fed animals had lower blood urea nitrogen (BUN) levels (P < 0.05), higher (P < 0.01) alanine aminotransferase (ALT) and lower serum iron (P < 0.001) than those fed Rhodesgrass hay based diets. There was a trend of increasing blood, leukocytes and specific gravity in the urine of NCF-fed animals. This experiment implies that feeding low quality non-conventional feeds containing antinutritional factors for relatively long periods might produce subtle negative effects on the physiology and chemistry of the digestive system and blood parameters which might negatively affect sheep health and make them more susceptible to diseases.     

The impact of weight loss on the 24-h profile of circulating peptide YY and its association with 24-h ghrelin in normal weight premenopausal women

Peptide YY (PYY) and ghrelin exhibit a reciprocal association and antagonistic physiological effects in the peripheral circulation. Research has yet to clarify the effect of weight loss on the 24 h profile of PYY or its association to 24 h ghrelin. We sought to determine if diet- and exercise-induced weight loss affects the 24 h profile of PYY and its association with 24 h ghrelin in normal weight, premenopausal women. Participants (n = 13) were assessed at baseline (BL) and after a 3-month diet and exercise intervention (post). Blood samples obtained q10 min for 24 h were assayed for total PYY and total ghrelin q60 min from 0800 to 1000 h and 2000 to 0800 h and q20 min from 1000 to 2000 h. The ghrelin/PYY ratio was used as an index of hormonal exposure. Statistical analyses included paired t-tests and linear mixed effects modeling. Body weight (−1.85 ± 0.67 kg; p = 0.02), and body fat (−2.53 ± 0.83%; p = 0.01) decreased from BL to post. Ghrelin AUC (5252 ± 2177 pg/ml/24 h; p = 0.03), 24 h mean (216 ± 90 pg/ml; p = 0.03) and peak (300 ± 134 pg/ml; p = 0.047) increased from BL to post. No change occurred in PYY AUC (88.2 ± 163.7 pg/ml; p = 0.60), 24 h mean (4.8 ± 6.9 pg/ml; p = 0.50) or peak (3.6 ± 6.4 pg/ml; p = 0.58). The 24 h association between PYY and ghrelin at baseline (p = 0.04) was weakened at post (p = 0.14); however, the ghrelin/PYY lunch ratio increased (p = 0.01) indicating the potential for ghrelin predominance over PYY in the circulation. PYY and ghrelin are reciprocally associated during a period of weight stability, but not following weight loss. An “uncoupling” may have occurred, particularly at lunch, due to factors that modulate ghrelin in response to weight loss.