Ruminal starch digestion characteristics in vitro of barley cultivars with varying amylose content

Three experiments were conducted to study effects of amylose/amylopectin ratios and starch particle size on ruminal digestion characteristics of barley starch using an automated in vitro gas production system. In Experiment 1, starch digestion characteristics were measured in 12 barley cultivars with different amylose/amylopectin ratios, both as milled grain and as purified starch isolated from the original grain samples. The same 12 cultivars, harvested 1 year later from the same locations, were used in Experiment 2. Gas production was measured in milled samples, and in neutral detergent fibre (NDF) extracted from the same samples. The objective of this approach was to estimate gas production from neutral detergent solubles (NDS) as an approximation of starch. This was done by subtracting the NDF gas curve from the total gas production curve. In Experiment 3, starch digestion characteristics were measured for large and small starch granules from nine of the original cultivars used in Experiment 1. The gas curves obtained were fitted to a three-pool Gompertz model, and the effective rate of digestion (kd) was estimated with a two-compartmental rumen model. In Experiment 1, the effective starch kd for milled barley and purified starch were 0.122 and 0.118/h, respectively. Barley cultivars with low amylose (LA) had a higher effective kd (0.148/h) compared with cultivars with normal amylose (NA) (0.115/h) and high amylose (HA) (0.102/h) (P=0.010). Results obtained with milled barley were supported by the purified barley starch sample results, but differences were smaller and only numerically different. In Experiment 2, the ranking of the amylose groups was consistent with those in Experiment 1 (i.e., LA > NA > HA) (P=0.096). However, these differences were not reflected in the effective kd for the NDS fraction (P=0.366). Thus, factors other than those related to starch per se, or other structural features, are apparently important. Barley cultivars in the LA group had a higher effective kd for aNDF (0.098/h) than did NA and HA barley (0.060 and 0.055/h, respectively). Thus, the effect of the amylose group on the effective kd for aNDF corresponded well with the milled barley results. The NDF fraction, directly or indirectly, has a clear impact on the ruminal digestion rate of barley starch. There was no difference in the effective kd for starch between the small (0.126/h) and large (0.129/h) starch granules.     

Rumen enzyme profile and fermentation characteristics in sheep as affected by treatment with sodium lauryl sulfate as defaunating agent and presence of ciliate protozoa

The efficiency of sodium lauryl sulfate as a defaunating agent and effect of rumen protozoa on nutrient utilization, fermentation characteristics and enzyme profile were evaluated in adult sheep maintained on a mixed ration containing 65:35% Pala (Ziziphus numularia) leaf: concentrate. Twenty-one adult Malpura sheep divided into three equal groups (DF, RF and F) were either defaunated by oral administration of sodium lauryl sulfate at the rate of 8 g/100 kg body weight (DF), or defaunated and again refaunated (RF), or maintained faunated (F). Daily dry matter intake was similar in defaunated, refaunated and faunated sheep. However, digestibility of cell wall and cell wall contents (NDF, ADF and cellulose) were lower (P < 0.01) in defaunated than refaunated and faunated sheep. Irrespective of the presence or absence of rumen protozoa, daily intake of DCP and DE were similar in the three experimental groups. Even with similar DM, DCP and DE intake, N-retention, blood glucose level, ruminal concentration of total VFA and total-N were higher (P < 0.01), while rumen pH and NH₃-N concentration were lower (P < 0.01) in defaunated sheep. Ruminal activity of amylase, xylanase, protease and urease enzymes were not influenced by presence or absence of ciliate protozoa. However, carboxymethyl cellulase enzyme activity was lower (P < 0.01) in the rumen of defaunated sheep. The total and differential counts of rumen protozoa were similar in refaunated and faunated sheep indicating lack of residual toxic effect of sodium lauryl sulfate. It is concluded that absence of ciliate protozoa increased ruminal TVFA, total-N with lower NH₃-N concentration and fibre digestibility in sheep. Moreover, sodium lauryl sulfate was fully effective for complete removal of rumen ciliate protozoa and successfully defaunated the sheep.     

Feed digestion,rumen fermentation and blood biochemical constituents in Malpura rams fed a complete feed-block diet with the inclusion of tree leaves

This experiment assayed the influence of the inclusion of dried Azardirachta indica, Albizzia lebbek or Ailanthus excelsa leaves in pearl millet stover-based complete feed block diets on feed intake, nutrient utilization, rumen fermentation characteristics, ciliate protozoa population and blood biochemical constituents in adult Malpura sheep. Complete feed blocks were formulated to have roughage-to-concentrate ratio of 70:30. Pearl millet stover (PMS) was used as basal roughage; 30 parts of pearl millet stover was replaced with dried leaves either of Azardirachta indica (NL), Albizzia lebbek (SL) or Ailanthus excelsa (AL). Twelve hogget Malpura rams, divided into four equal groups, were offered one of the four dietary treatments. A feeding-cum-metabolic trial was conducted to assess nutrient utilization. Rumen liquor samples were collected at 0, 3, 6, 12, 18 and 24 h post-feeding to assess rumen fermentation pattern and ciliate protozoa population. Inclusion of dried leaves in PMS-based diets improved CP and DCP content. Dietary DCP was low (P < 0.01) in PMS (8.52%) compared to tree leaves (9.77–11.59%) diets. AL and NL diets had higher (P < 0.05) DCP than the SL diet. The inclusion of tree leaves did not influence organic matter, crude protein or cellulose digestibility, but depressed dry matter, NDF, ADF and energy digestibility. DE content was also lower in tree leave diets. Inclusion of tree leaves improved CP and DCP intake, but DE intake and nitrogen utilization did not change. The pH of rumen liquor (SRL) was low (6.99, P < 0.05), but total nitrogen (52.9 mg/dl SRL) and NH₃-nitrogen (9.34 mg/dl SRL) concentrations were higher (P < 0.01) in the AL diet. TVFA concentrations and ciliate protozoa population were similar on the four diets. Animals in the four groups had the desired concentration of rumen metabolites required for fibrous diets. Complete feed-block feeding provided a constant nutrient supply to rumen microbes that optimise rumen fermentation. Blood biochemical constituents did not change due to the inclusion of tree leaves. Therefore, tree leaves can be included with roughage-based feeding to improve the protein nutrition status of ruminants. Further studies are required to assess the negative influence of tree leaves on digestibility.     

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.     

Evaluation of a two-stage in vitro technique for estimating digestibility of equine feeds using horse faeces as the source of microbial inoculum

To develop a two-stage in vitro technique that simulates both pre-caecal and hind gut digestion processes, four enzymatic pre-digestion treatments by pepsin and α-amylase (ET0 = control, ET1 = 2 h pepsin + 2 h amylase, ET2 = 2 h pepsin + 4 h amylase, ET3 = 8 h pepsin + 16 h amylase) were tested on oat hay (OH), barley grain (BG) and soybean meal (SBM). Investigated parameters were enzymatic organic matter digestibility (OMDe), and gas production (G48h, G72h) and OM digestibility (OMD) using horse faeces as a source of microbial inoculum.Enzymatic pre-digestion treatments affected (P<0.05) investigated parameters and their ranking differed among feeds. Only OMD of BG and SBM were higher after the pre-digestion treatment. OMD prior to (ET0) and after ET3 application were, successively, 0.357 versus 0.351 (OH), 0.71 versus 0.79 (BG) and 0.70 versus 0.78 (SBM). Net gas production overestimated fermentation potential of non-pre-digested feeds. G72h (ml/g DM) prior to (ET0) and after ET3 application were, successively, 80.3 versus 58.0 (OH), 151.7 versus 30.4 (BG) and 110.6 versus 37.7 (SBM).It was concluded that the enzymatic pre-digestion treatments effects varied among tested feeds, and that the suggested procedure be extended and validated with a large array of feeds of known digestibility values.     

Effect of harvesting period on the nutritive value of rice grass (Echinochloa sp.) hay given as sole diet to lambs

Data regarding the influence of maturity within the vegetative stage of tropical grasses on forage quality are limited and conflicting. The change in chemical composition of rice grass (Echinochloa sp.) hay harvested at 32, 46, 72 and 90 days of regrowth, and its effect on intake, digestibility, ruminal fermentation, rumen microbial protein synthesis (Experiment 1) and splanchnic oxygen uptake (Experiment 2) by lambs was evaluated. Except intake of indigestible neutral detergent fibre (NDF) which was similar for all treatments, intake of all hay components and the apparent digestibility of dry matter, organic matter (OM), NDF, N, as well as OM and N true digestibility, N retention and rumen microbial protein synthesis decreased linearly (P < 0.05) with increased regrowth age. Rumen fluid pH, ammonia N and peptide concentrations were similar for all treatments while sugars and amino acid concentrations decreased linearly with increased regrowth age of rice grass (P < 0.05). Passage rate of particles through reticulum-rumen (PRrr) was quadratically related (P < 0.05) to regrowth age. The highest PRrr and, consequently, the lowest retention time in the reticulum-rumen were obtained at 72 days of regrowth. There was a quadratic effect (P < 0.05) on net portal-drained viscera (PDV) flux of oxygen and heat production, while OM intake, portal blood flow and heat production as proportion of digestible energy (DE) intake were not affected by the increased regrowth age of rice grass. The highest means of oxygen uptake and heat production by PDV tissues were in 72 days treatment. In the whole splanchnic metabolism assay neither hay intake nor blood flow, oxygen uptake or heat production were affected by forage regrowth age. In conclusion, the nutritive value of rice grass hay decreased as regrowth age increased from 32 to 90 days due to decrease both OM intake and digestibility.     

Nutrient composition,digestion and rumen fermentation in sheep of wheat straw treated with calcium oxide,sodium hydroxide and alkaline hydrogen peroxide

This study tested the effect of calcium oxide (CaO), sodium hydroxide (NaOH) and NaOH plus hydrogen peroxide (H₂O₂; AHP) on cell wall composition, digestion and fermentation of wheat straw (straw) in sheep. Treated straws were prepared by mixing straw either with water followed by dusting with CaO at 160 g kg⁻¹ DM or with a NaOH solution alone at 3 l kg⁻¹ DM to supply 80 g NaOH kg⁻¹ DM (Na) or pre-soaked with Na exactly as in the previous treatment for 27 h followed by mixing with 130 g H₂O₂ kg⁻¹ DM (AHP) for 6 h. After 14 days of storage, the treated straws and an untreated straw (U) were fed automatically every 2 h to four individually housed sheep together with a supplement in a 4×4 latin square experiment. Each kilogram supplement DM contained 422 g CP and 10.8 MJ ME. NDF (p<0.001) and hemicellulose (p<0.01) contents were significantly reduced whereas cellulose was increased (p<0.001) in treated compared to untreated straw. ADL was reduced in Ca (p<0.05) but increased (p<0.05) in Na and AHP compared with U. The rumen and total tract digestibility were significantly (p<0.001) greater in sheep fed treated compared with untreated straw. Significant differences (p<0.05) between treatments for pH, NH₃ and VFA were also observed. All treatments improved the nutritive value of straws compared with untreated through modification of cell wall with a subsequent increase in digestibility by sheep. Although the digestibility for Ca was lower than that for Na despite reduction in cell wall, its use to treat straws may be more safe and cost effective than Na. AHP was the most effective and could also improve the energy value of other low quality forages for ruminants. However, the need of AHP for a large amount of NaOH to achieve highly alkaline pH limits its farm scale application. Therefore, further studies should either consider reducing the amount of NaOH or finding alternative alkalis that are cost effective and user-friendly.     

Effects of urea and aqueous ammonia treatment on the composition and nutritive value of maize residues

Two trials were conducted to investigate changes in chemical composition and nutritive value of maize residues treated with urea and aqueous ammonia. In Experiment I, maize stalks, husks and cobs were chopped into pieces of approximately 1 cm length. Aqueous NH₃ (3%) and feed grade urea equivalent to 3% NH₃ were sprayed into 100 g duplicate samples on DM basis, thoroughly mixed, stored in plastic containers and kept for 3 week at ambient temperature (25–27 °C). Proximate composition and in vitro dry matter digestibility of test materials were determined. In Experiment II, 100 kg batches of the maize residues were chopped and treated as in Experiment I but stored in 205 l drums lined with black polythene sheet. Feed intake and nutrient digestibility of treated materials were evaluated with 18 mature WAD sheep averaging 19.8 kg BW. Treatment improved (P < 0.05) intake and digestion coefficients for N, DM, NDF, ADF and OM but there were no differences (P > 0.05) between aqueous NH₃ and urea treatments. Feed grade urea or the equivalent weight of fertilizer grade urea can be used to improve the nutritional value of maize residues for small ruminant feeding during off season periods.     

Effect of elemental nano-selenium on feed digestibility,rumen fermentation,and purine derivatives in sheep

The objective of this experiment was to study the effect of elemental nano-selenium (NS) on feed digestibility, rumen fermentation, and urinary purine derivatives in sheep. Eight male ruminally cannulated sheep (42.5 ± 3.2 kg of body weight, BW) were used in a replicated 4×4 Latin square experiment in four 20 day periods. Depending on treatment designation, sheep were fed the basal diet supplemented with 0 (control), 0.3, 3 and 6 g of nano-Se/kg dry matter (DM). Ruminal pH (range of 6.68–6.80) and ammonia N concentration (range of 9.95–12.49 mg/100 mL) was decreased (P<0.01), and total VFA concentration (range of 73.63–77.72 mM) was increased linearly (P<0.01) and quadratically (P<0.01) with increasing nano-Se supplementation. The ratio of acetate to propionate was linearly (P<0.01) and quadratically (P<0.01) decreased due to the increasing of propionate concentration. In situ ruminal neutral detergent fiber (aNDF) degradation of Leymus chinensis and crude protein (CP) of soybean meal were linearly (P<0.01) and quadratically (P<0.01) improved by feeding nano-Se. Similarly, nutrients digestibility in the total tract and urinary excretion of purine derivatives were also quadratically (P<0.01) changed by increasing nano-Se supplementation. The present results indicated that nano-Se supplementation in basal diet improved rumen fermentation and feed utilization. Nano-Se could also stimulate rumen microbial activity, digestive microorganisms or enzyme activity. The optimum dose of nano-Se was about 3.0 g/kg dietary DM in sheep.     

Influences of flavomycin,ropadiar, and saponin on nutrient digestibility,rumen fermentation,and methane emission from sheep

This study focused on the effects of three additives given together with a hay/concentrate-based diet on nutrient digestibility, rumen fermentation, and methane emission from sheep. The basal diet consisted of 1.29 kg mixed hay and 0.43 kg concentrate mixture based on dry matter (DM). Treatments consisted of control (no additive), flavomycin⁴⁰ (250 mg/d), ropadiar from an oregano extract (250 mg/d), and saponin in the form of a yucca schidigera extract (170 mg/d). Results indicated that intake and digestibility were unaffected by treatments (P>0.05). The NH₃-N concentration of rumen liquor was lower (P<0.05) for additive treatments versus the control treatment. Higher concentrations of volatile fatty acid (VFA) were observed in the saponin (75.8 mmol/L) and ropadiar (73.1 mmol/L) treatments. The proportion of individual fatty acid of rumen liquor was unchanged, whereas lower ratio of acetate to propionate in the saponin treatment was observed (P<0.05). The average methane production expressed on digested organic matter (OM) and neutral detergent fiber (aNDFom) basis were decreased by approximately 3.3 and 12.0 g/kg, respectively in saponin, and 4.2 and 11.9 g/kg in ropadiar treatment compared to the control. Methane production was positively correlated with the concentrations of NH₃-N, and negatively correlated with total VFA and the proportion of propionate of rumen liquor (P<0.05). The study found that saponin and ropadiar could have the potential to reduce rumen methanogenesis in sheep.     

Effects of sodium bicarbonate and yeast on nutrient intake,digestibility, and ruminal fermentation of light-weight lambs fed finishing diets

This study was conducted to determine the effects of adding yeast culture and sodium bicarbonate to the finishing diets for lambs on intake, digestibility, rumina1 parameters and nitrogen retention. Twenty Pelibuey male lambs, weighing an average of 23 kg, were assigned to one of four treatment groups according to a completely randomized design with a 2 × 2 factorial arrangement. Treatments were: (1) no additive (NA); (2) 0.12% yeast culture (YC); (3) 0.5% sodium bicarbonate (SB); and (4) 0.12% YC and 0.5% SB. During the 7-day collection period, SB increased DM intake (P < 0.05), while YC had no effect (P > 0.05) on intake. Intake of non-fibrous carbohydrates (NFC) was increased (P < 0.05) with SB in the ration, but not by supplementing YC (P > 0.05). The YC had no effect (P > 0.05) on dry matter (DM), neutral detergent fiber (NDF) or non-fibrous carbohydrates digestibility. A higher intake of NFC with the SB treatments was associated with a lower (P < 0.05) digestibility. SB increased excretion of NDF (P < 0.05), which reduced its digestibility (P < 0.05) by more than 9 percentage units. Lambs consuming diets with SB had 27% more N retained, in contrast with those fed the basal diet without additives. Rumen pH was greater than the minimum considered to cause acidosis (≤pH 5.5). The SB reduced (P < 0.05) percent molar acetate and increased (P < 0.05) percent molar propionate, which is in contrast to what has been normally observed with dairy cattle research. A lower acetate to propionate ratio with the SB rations may be a result of a lower digestibility and a higher intake of NDF, which may have increased the rate of passage of fiber particles through the gastrointestinal tract.     

Effect of rare earth elements on in vitro rumen microbial fermentation and feed digestion

The objectives of this study were to investigate the effects of rare earth elements (REEs) on in vitro rumen fermentation, gas production, microbial protein synthesis and nutrient digestion using in vitro batch culture and continuous culture technique. A mixture of REE containing (g/kg) 380 g of LaCI₃·6H₂O, 521 g of CeCI₃·6H₂O, 30 g of PrCI₃·6H₂O and 69 g chlorides of other light REEs. The experimental diet consisted of 885 g/kg barley grain, 84 g/kg barley silage and 31 g/kg supplement (dry matter (DM) basis). Diet supplemented with different dosages of REE (control, no additional REE; low, 400 mg/kg REE; and high, 800 mg/kg REE, DM basis) were incubated for 4, 8, 14 and 24 h in diluted rumen fluid. At the end of 24 h of incubation, gas production and concentration of volatile fatty acid (VFA) linearly increased with increasing REE supplementation; whereas, influence of REE supplementation on VFA profile was marginal. Dry matter disappearance was not affected (P>0.10). Six dual-flow continuous culture fermenters were used in a replicated 3 × 3 Latin square with same treatments and same diet used in the batch culture. Mean ruminal pH (5.71) and total VFA (93.6 mM) concentration were not affected by supplementation of REE. The molar proportion (mol/100 mol) of acetate (39.1) and propionate (50.5) was similar among the treatments. However, the proportion (mol/100 mol) of butyrate was higher with the high REE (6.6) than with low REE (5.3) or the control (5.8). Ruminal true digestibilities of organic matter (OM) (0.785, 0.811 and 0.828), acid detergent fibre (0.360, 0.431 and 0.432) and crude protein (0.496, 0.590 and 0.589) for control, low and high REE, respectively, linearly increased with increasing REE supplementation, whereas, the increase in ruminal digestibility from low to high dosage of REE was minimal. Microbial nitrogen (N) production (g/day) and microbial efficiency (g N/kg of truly fermented OM) were not affected by treatments. Improvement of ruminal digestibility of OM due to REE supplementation was attributed to the increase in digestibility of fibre and degradability of protein. The results suggest that REE supplementation improved ruminal fibrolytic and proteolytic activities.     

Interpretation of rumen degradability of concentrate feeds with a Gompertz model

An investigation of the kinetics and synchronicity of rumen crude protein and starch degradability was performed for maize, flaked maize, ensiled maize cob, barley, flaked barley, wheat, oat, sorghum and triticale grain, using the in situ polyester bag technique. Kinetics of rumen degradability were corrected for particle losses from the bag before degradation. Washing losses were measured by shaking feed samples in polyester bags in tap water at 20°C for 1 h and recovering the particle losses in fibre glass filter. Mean washing losses of dry matter were 442 g/kg DM (soluble fraction 17%); mean washing losses of nitrogen were 446 g/kg nitrogen, with 52% being water soluble. Starch escaped to a higher extent from the bags in the washing machine (average washing losses were 581 g/kg starch, with a soluble fraction of 7%). Degradability data for dry matter, nitrogen and starch measured at each time of incubation were corrected for the respective particle losses and fitted with both first order and Gompertz (sigmoidal) models. The difference between the estimated parameters obtained with the two models was negligible, although differences occurred for the immediately soluble fraction (a) of dry matter and starch and for total degradable fractions of dry matter, nitrogen and starch. No differences were observed between effective degradabilities, independent of rumen outflow rate (0.04 and 0.08/h), apart for the mean effective degradability of nitrogen at 0.08/h, which was higher for the first order model (577 versus 564 g/kg, P < 0.001). The advantage of using the Gompertz model to interpret the kinetics of rumen degradability of cereals was due to the possibility of studying synchronicity of nitrogen and starch released into the rumen and ranking feeds according to their degradability pattern, which required the calculation of the first derivatives of the Gompertz model for nitrogen and starch and weighted for their respective amounts in the feeds. The difference of these ratios from the optimum value of 30 mg N per 1 g starch gives the instantaneous synchronicity of the cereals. Maize, flaked maize, maize cob and sorghum had negative and almost constant values during the first 8 h of incubation. Barley, flaked barley, wheat, triticale and oat changed from an initially negative value to a positive value from about 4 h onwards; oats had a similar pattern, but a positive value was observed only after 6 h.     

Effects of feed intake on enteric methane emissions from sheep fed fresh white clover (Trifolium repens) and perennial ryegrass (Lolium perenne) forages

Published analyses of enteric methane (CH₄) emissions from sheep and cattle show an inverse relationship between feed intake and CH₄ yield (g CH₄/kg dry matter (DM) intake), which suggests opportunities for reducing CH₄ emissions from feed eaten and per unit of animal production. Most relationships between feed intake and CH₄ yield have been based on animals fed conserved feeds, especially silages and grains. Our research is a series of experiments with fresh white clover (Trifolium repens) and perennial ryegrass (Lolium perenne; ryegrass) forages fed to sheep at a range of feed intake levels. This study was comprised of four experiments where good quality freshly harvested white clover or ryegrass were fed to sheep over a three-fold range in DM intake, and CH₄ emissions were measured in respiration chambers for two consecutive days in each experiment. Measurements were made from 16 sheep in Experiment 1 (fed at 1.6 × metabolizable energy requirements for maintenance; ME_m), 28 sheep in Experiment 2 (at 0.8 and 2.0 × ME_m), eight sheep and two measurement periods in Experiment 3 (at 1.6 × ME_m), and 30 sheep in Experiment 4 (fed at 0.8, 1.2, 1.6, 2.0 and 2.5 × ME_m). Prior to each experiment, sheep had a 10 d acclimatization period to diets. Apparent digestibility was measured over 7 d from sheep in Experiments 1, 3 and 4, along with collection of rumen digesta for volatile fatty acid (VFA) determination. Although CH₄ yields differed when sheep were fed white clover or ryegrass at similar intakes, the differences were inconsistent and mean values similar across all experiments. This, and a similar structure of all experiments, enabled combined analysis of data from all four experiments using the restricted maximum likelihood (REML) procedure to estimate effects of feed intake level on digestibility, digestible nutrient intake, gas emissions, and VFA concentrations in the rumen. The REML analysis showed that when DM intake increased from 0.40 to 1.60 kg/d, the predicted responses were an increase in CH₄ production (g/d) of 187% (12.4–35.6 g/d; P<0.001), and a decline in CH₄ yield of 21% (25.6–20.2 g/kg DM intake; P<0.001). High feed intake levels were associated with increased molar proportions (mM of total VFA) of propionate from 0.17 to 0.21 (P=0.038). Single and multiple regressions were completed on the data from all experiments, with organic matter (OM) intake predicting 0.87 of the variation in CH₄ production, and molar proportion of propionate predicting 0.60 of the variation in CH₄ yield. Increasing feed intakes by 1 kg/d of DM reduced CH₄ yield by 4.5 g/kg DM intake. Plant chemical composition was weakly related to CH₄ yield. High intakes of fresh forages will lower CH₄ yield from fermentation, but effects of feed composition on CH₄ emissions were minor. The interaction between effects of feed intake and rumen function requires further investigation to understand relationships with CH₄ emissions.     

Ruminal large and small particle kinetics in dairy cows fed red clover and grass silages harvested at two stages of growth

Passage, comminution and digestion rates of large and small particles were estimated using a rumen evacuation technique and total faecal collection with five lactating dairy cows in a 5 × 5 Latin square experiment. Two grass and two red clover silages harvested at early and late primary growth stages and a 1:1 mixture of late harvest grass and early harvest red clover were the dietary treatments. Cows received 9.0 kg supplementary concentrate per day. Ruminal contents and faeces were divided into large (>1.25 mm) and small (1.25–0.038 mm) particles by wet sieving. Indigestible neutral detergent fibre (iNDF) was determined by 12 days ruminal in situ incubation followed by neutral detergent extraction. Plant species did not affect ruminal particle size distribution, whereas advancing forage maturity decreased the proportion of large particles for both grass and red clover silage diets. Ruminal pool size of iNDF was higher (P<0.001) with red clover compared to grass silage diets. Ruminal passage rates of iNDF and potentially digestible NDF (pdNDF) increased with decreasing particle size (P<0.01). Passage rate of iNDF for small particles was slower (P<0.01) when red clover compared to grass silage diets were fed. Particle comminution rate in the rumen was slower (P<0.001) with red clover compared to grass silage diets and it increased (P<0.01) with advancing forage maturity. The contribution of particle comminution to ruminal mean retention time of iNDF in the ruminal large particle pool was smaller (P<0.01) in red clover compared to grass silage diets and it increased (P<0.05) with the mixed silage compared to the separate silages. Passage rate of pdNDF for both large and small particles was not affected by dietary treatments. Digestion rate of pdNDF for large particles was faster (P<0.001) with red clover compared to grass silage diets. Differences in ruminal passage and digestion rates of the large and small particles, in addition to differences in the passage and digestion rates of red clover compared to grass silage diets, emphasize the need to consider particle size and forage type in metabolic models predicting feed intake and fibre digestibility in ruminants.     

In vivo and in situ digestibilities and nitrogen balance of Buddleia skutchii as a sole component and mixed with Pennisetum clandestinum in sheep diets

The objective of this work was to evaluate the intake (I) of all Buddleia skutchii foliage (BSF), collected from three different sites (places: A, B and C) in feeding sheep (phase 1), and in combination with Pennisetum clandestinum (Pc) (phase 2). Trials of feed intake, in vivo dry matter digestibility (IVDMD), and digestibilities of crude protein (CP), acid detergent fiber (ADF), neutral detergent fiber (NDF), and nitrogen balance were carried out during phase 1. Diet was given to nine Creole male lambs of 23(±3) kg body weight (BW), located in individual metabolism crates. During phase 2, four treatments were evaluated (T1 = 100% Pc, T2 = 20% BSF + 80% Pc, T3 = 40% BSF + 60% Pc and T4 = 60% BSF + 40% Pc) in a 4 × 4 Latin square design, using four ruminal cannulated Creole male rams of 50 ± 2 kg BW and remained in individual metabolism crates. In vivo digestibility (DM, CP and gross energy (GE)) and kinetics of in situ disappearance of DM were evaluated. The results in phase 1 indicated that BSF from Site B had relatively higher CP (14.9%), ADF (50.6%) content and nitrogen retention (1.93 g d⁻¹). On the other hand, the lambs showed intake levels of 376.29 g d⁻¹ DM, 56.21 g d⁻¹ CP, 206.24 g d⁻¹ ADF and 1418.21 kcal gross energy (P > 0.05); and digestibilities of 54.23% DM, 47.17% CP and 39.31% NDF. Therefore, BSF from Site B was selected for the second phase of this work as the best result. We observed that the nutrition quality of the diets was higher as a result of increasing BSF concentration in the diet. When sheep were fed on 40% BSF–60% Pc, DM intake levels of 900.6 g d⁻¹, and DM, CP and GE digestibility of 70.0, 52.82 and 55.1%, respectively, were observed. There were not significant differences among diets, and in situ disappearance, K_d and K_p, but the diet with 60% of BSF presented higher ruminal digestibility (42.3%). The results allowed to validate the local knowledge on BSF as a promising forage species, and showed that BSF should be incorporated in a sheep production system.     

In situ degradability of seven plant protein supplements in heifers fed high concentrate diets with different forage to concentrate ratio

Four Holstein heifers (297.5 ± 27.7 kg BW) fed high concentrate diets were used in a crossover experiment in order to characterize the rumen fermentation pattern, and to estimate by the in situ method rumen degradation kinetics of alfalfa hay and seven plant protein supplements: solvent-extracted soybean meal, solvent-extracted sunflower meal, peas (Pisum sativum L.), lupin seeds (Lupinus sp.), broadbean (Vicia faba L.), horsebean (Vicia faba L. var equina) and vetch (Vicia sativa L.), in high concentrate diets with different forage to concentrate ratio. Heifers were fitted with a ruminal cannula. The experiment was performed in two 30-day periods, 15 days of diet adaptation and 15 days of sampling. At each period, heifers were offered one of two total mixed rations (12:88 versus 30:70 forage to concentrate ratio), two heifers per diet, on ad libitum basis. After the first period, heifers switched treatments. Intake of dry matter (DM), organic matter, crude protein and neutral detergent fibre (NDF), expressed as kg/day, did not differ between treatments, but DM intake, expressed as g/kg metabolic body weight (BW), was higher in the 12:88 diet. Average rumen pH was 6.0 in both diets, and the time pH was below 5.8, which is considered as a critical threshold for fibre degradation, was the same for both treatments (10.4 ± 1.6 h). Average ammonia nitrogen and volatile fatty acid (VFA) concentrations did not differ between treatments and individual VFA proportions were typical of high concentrate diets. Average effective degradability of DM (0.62 ± 0.02) and NDF (0.25 ± 0.03) of alfalfa hay were low and no differences were detected between treatments. The same extent of NDF degradation, together with the same proportions of VFA would indicate that both diets had the same fibrolytic activity. Forage to concentrate ratio did not affect rumen nitrogen degradability of any protein supplements incubated in situ. Corrected effective degradability for small particle losses of sunflower meal (0.78) was higher than legume seeds, which were not statistically different between each other and ranged from 0.63 to 0.66. Soybean meal had the lowest degradability value (0.61). These nitrogen degradation values must be considered more valid for beef cattle formulation of high concentrate diets than data obtained with forage diets.     

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.     

Effect of dietary cobalt supplementation on plasma and rumen metabolites in Mehraban lambs

Two experiments were conducted to study the effects of different levels of dietary cobalt on performance, plasma and rumen metabolites and nutrient digestibility in Mehraban male lambs. Experiment 1: 28, 8–9-month-old lambs were randomly divided into four groups. Animals were fed a basal diet containing 0.088 mg Co/kg DM and were supplied with 0 (control), 0.25, 0.50, or 1.00 mg Co/kg DM as reagent grade CoSO₄·7H₂O. The experiment lasted for 70 days. Experiment 2: four lambs from each group in Experiment 1 were randomly allocated to the individual metabolic crates to measure the effects of dietary Co on nutrient digestibility. Final body weight, average daily gain and gain efficiency were higher (p < 0.05) in the group supplemented with 0.50 mg Co/kg DM compared to other groups. Plasma glucose and vitamin B₁₂ concentrations increased (p < 0.05) at all levels of Co supplementation on day 68 of the experiment and for vitamin B₁₂ were higher (p < 0.05) at 0.50 and 1.00 mg Co/kg DM compared to 0.25 mg Co/kg DM. There was no significant difference among treatments for TVFA and ruminal fluid pH. Digestibility of dry matter, organic matter, crude protein and neutral detergent fiber increased (p < 0.05) by Co supplementation, but did not differ among Co supplied treatments. The obtained results showed that lambs fed the control diet containing 0.088 mg Co/kg DM had a reduced appetite and gained less than the supplemented animals, suggesting that the level of 0.088 mg Co/kg DM was inadequate for normal growth of Mehraban male lambs, and a total level of 0.58 mg Co/kg DM might be optimum level for enhancing performance.     

Comparison of bovine rumen liquor and bovine faeces as inoculum for an in vitro gas production technique for evaluating forages

Two experiments were undertaken using the in vitro gas production technique of Theodorou et al. [Anim. Feed Sci. Technol. 48 (1994) 185] to compare rumen liquor (RL) and faeces (FA) as inocula for fermenting gramminaceous forages over 96 h periods. Experiment 1 used 12 forages of differing in vivo digestibility (ammonia treated wheat straw, field-cured hay (Lolium perenne) and 10 artificially dried grasses (L. perenne) harvested at different maturities). Experiment 2 used seven maize-silage based forages (whole plant, stover, leaf, lower stem, middle stem, upper stem and husk). In both experiments, rumen liquor and faeces were obtained from two cows in early lactation, each fed daily with 9.4 kg DM of grass silage and 9.0 kg DM of concentrate. Rumen contents were sampled through the fistula, before morning feeding; faeces were sampled from the rectum, immediately afterwards. Rumen liquor (250 ml) was prepared by straining contents through two layers of muslin, adding the solids after blending with 250 ml of buffer and re-straining. Faeces were prepared by mixing (300 ml) with 150 ml of buffer and straining through two layers of muslin and adding a homogenate of the solids and 150 ml of buffer after straining. Data were fitted to the model of France et al. [J. Theor. Biol. 163 (1993) 99]. All model parameters showed FA to have a poorer fermentation capacity than RL. In both experiments, potential gas production volumes (A) were lower (on average 52.9 ml (18.5%)) and lag times longer (on average 2.9 h) for FA compared to RL. Fractional rate of fermentation at half asymptote (T/2) was generally greater for RL than FA (overall means, 0.042, 0.028) and the time required to T/2 being less (overall means, 21.9, 35.4 h). However, potential gas production (A) was highly correlated between RL and FA: Experiment 1 (r²=0.94, 11 forages, excluding ammonia treated straw) and Experiment 2 (r²=0.83, six forages, excluding middle stem). In Experiment 1, organic matter digestibility in vivo (OMD_IV) was also highly correlated with both OMD_FA (r²=0.77, 11 forages) and OMD_RL (r²=0.89, 11 forages); OMD_RL and OMD_FA were also highly correlated (r²=0.81). Similar correlations occurred in Experiment 2. It is concluded that faeces have potential as an alternative inoculum to rumen liquor for in vitro gas production techniques, but methods of overcoming the longer lag phase with faeces require further research.