Comparison of biological and chemical methods,and internal and external standards,for assaying tannins in Spanish shrub species

Tannin contents of stems, flowers and leaves of four shrub species (Erica arborea, Erica australis, Cytisus cantabricus and Genista occidentalis) were chemically assessed using two colorimetric assays (butanol–HCl and Folin–Ciocalteu) and several standards (tannic acid, purified quebracho condensed tannins (CT) and purified CT extracted from each species). A method based on the biological activity of tannins in the rumen was also used for assaying tannins. Depending on the species, the use of quebracho tannin as standard over‐ or underestimated CT contents, in comparison to the use of the respective internal standard. However, the high correlationship observed between in vitro gas production in the presence or absence of polyethylene glycol (PEG) and CT contents estimated from quebracho suggests that this standard could be used for a quicker estimate of the nutritive value of tannin‐containing plants. Tannin contents obtained by the Folin–Ciocalteu method, using tannic acid as standard, were also related to their activity on rumen fermentation. The use of the in vitro gas production technique, in the absence or presence of PEG, may provide more useful information than the chemical assays when the final aim of tannin analysis is to assess the nutritive value of feedstuffs for ruminants. Copyright © 2004 Society of Chemical Industry     

In vitro effects of steroidal saponins from Yucca schidigera extract on rumen microbial protein synthesis and ruminal fermentation

In a first experiment, ground alfalfa hay and rolled barley grain were incubated in buffered ruminal fluid with and without Yucca schidigera extract (YE, 0 or 10 mg ml⁻¹). Gas and total VFA production from barley grain were increased (P < 0.05) by YE during the first 10 h of incubation; from alfalfa hay, these were reduced (P < 0.001) throughout the 24 h. Yucca extract reduced (P < 0.001) acetate/propionate ratios and ammonia concentrations, irrespective of substrate. In a second experiment, ground barley grain was incubated in a buffered suspension of mixed ruminal microbes obtained by low‐speed centrifugation of ruminal fluid. Steroidal saponins (SAP) isolated from YE were included at 0, 15, 75 or 225 µg ml⁻¹. Microbial incorporation of ¹⁵N was increased (P < 0.05) by 15 µg SAP ml⁻¹ but decreased (P < 0.05) by 225 µg SAP ml⁻¹. Gas and VFA production peaked with 75 µg SAP ml⁻¹ and were elevated (P < 0.05) relative to control in the presence of 75 µg SAP ml⁻¹. Microbial protein synthesis was increased (P < 0.05) by SAP at 15 µg ml⁻¹ and reduced (P < 0.05) by the higher concentrations. Acetate/propionate ratios were linearly reduced by SAP from 8 to 24 h incubation (P < 0.01). The effects of SAP on digestive microbes were less pronounced (P < 0.05) with barley grain digestion than with alfalfa. A YE × diet interaction was recorded. The biological activity of YE was shown to be attributable to its SAP and to be diet‐dependent. © 2000 Society of Chemical Industry. Contributions of Y Wang, T A McAllister, L J Yanke and Z Xu © Minister of Public Works and Government Services Canada 2000     

A comparison between buffalo (Bubalus bubalis) and cow (Bos taurus) rumen fluids in terms of the in vitro fermentation characteristics of three fibrous feedstuffs

Rumen fluids from fistulated buffalos (Italy‐BRF) and cows (Netherlands‐CRF) were used as inocula to determine the fermentation kinetics of three forages. These were corn silage (CS), grass silage (GS) and wheat straw (WS) which had originated from both regions, giving six substrates in total. Fermentation kinetics was assessed by the measurement of cumulative gas production. Organic matter (OM) loss and volatile fatty acid (VFA) concentration at the end of the fermentation period were also determined. Both BRF and CRF ranked the substrates in the same order for total VFA, total gas production and the maximum rate of substrate degradation (CS > GS > WS). However, while the ranking of substrates was the same for both species, the absolute values differed significantly between the two inocula. Gas production, expressed as cumulative volume per unit mass of incubated (OMCV, ml g^?1) and as cumulative volume per unit mass of OM degraded (OM ml g^?1) for CRF was consistently higher than that for BRF for all substrates (p < 0.0001). VFA production, particularly of acetic and butyric acids, was significantly (p < 0.0001) lower for BRF than for CRF, though the digestibility of OM was the same for both inocula. VFA production predicted by the use of stoichiometric equations was generally higher than the observed gas production. These equations also predicted that the calculated amount of OM utilised for microbial growth was higher for BRF than for CRF. This may explain why diets having the same energy content but less protein can be fed to buffalo since they seem to have a lower requirement for protein compared with cattle. This possibility will need to be investigated in vivo. Given the similar ranking of feedstuffs between the two sources of inocula, these results suggest that either inoculum would be suitable for use in the cumulative gas production test as a measure of feedstuff evaluation. However, given the differences in absolute values, it is recommended that inocula from the species which will receive the feedstuff should be used. Copyright © 2004 Society of Chemical Industry     

Evaluating the effects of Lactobacillus animalis and Propionibacterium freudenreichii on performance and rumen and fecal measures in lactating dairy cows

Two experiments evaluated the effect of supplementation with a bacterial direct-fed microbial on performance and apparent total-tract nutrient digestion of dairy cows. In experiment 1, 30 multiparous cows (75 ± 32 d in milk) were randomly assigned to 1 of 2 treatments fed for 10 wk. All cows were fed a diet containing 23.8% starch. Treatments were top dressed to rations twice daily and consisted of a combination of Lactobacillus animalis (1 × 10⁹ cfu/d) and Propionibacterium freudenreichii (2 × 10⁹ cfu/d; LAPF) or carrier alone (CON). In experiment 2, 6 ruminally cannulated cows (123 ± 129 d in milk) were randomly assigned to a crossover design with two 6-wk periods. Cows received the same CON or LAPF treatment as in experiment 1. Cows were fed the same 23.8% starch diet as experiment 1 during wk 1 through 5 of each period, and then cows were abruptly switched to a 31.1% starch diet for wk 6. For both experiments, intake and milk yield were measured daily, and milk samples were collected weekly. In experiment 1, fecal grab samples were collected every 6 h on d 7 of experimental wk 1, 2, 4, 6, 8, and 10. Fecal consistency was scored, and fecal starch was measured in daily composite samples. Fecal composites from a subset of 7 cows per treatment were used to measure apparent total-tract nutrient digestion. In experiment 2, rumen pH was continuously recorded during wk 5 and 6. On d 7 of wk 5 (the final day of feeding the 23.8% starch ration), d 1 of wk 6 (the day of diet transition), and d 7 of wk 6 (the final day of feeding the 31.1% starch ration), rumen in situ digestion was determined. Samples of rumen fluid and feces were collected every 6 h on those days for measurement of fecal starch (composited by cow within day), rumen volatile fatty acids, and fecal pH. Rumen and fecal samples were collected at one time point on those days for microbiota assessment. In experiment 1, treatment did not affect intake, milk yield, milk composition, or fecal score. The LAPF treatment decreased fecal starch percentage and tended to increase starch digestion compared with CON, but the differences were very small (0.59 vs. 0.78% and 98.74 vs. 98.46%, respectively). Digestion of other nutrients was unaffected. In experiment 2, LAPF increased rumen pH following the abrupt switch to the high-starch diet, but milk yield was lower for LAPF compared with CON (35.7 vs. 33.2 kg/d). Contrary to the decrease in fecal starch with LAPF observed in experiment 1, fecal starch tended to be increased by LAPF following the abrupt ration change in experiment 2 (2.97 vs. 2.15%). Few effects of treatment on rumen and fecal microbial populations were detectable. Under the conditions used in our experiments, addition of the bacterial direct-fed microbials did not have a marked effect on animal performance, ruminal measures, or total-tract nutrient digestion.     

Factors affecting in vitro formation of tannin-protein complexes

In interaction of condensed tannins from Desmodium intortum and Lotus pedunculatus and tannic acid (hydrolysable tannin) with salivary mucoproteins (from sheep and goats), plant leaf proteins and bovine serum albumin were evaluated. These studies were carried out over a pH range of 2-0-9-0 and different inorganic ion conditions to simulate conditions in which dietary proteins would interact with tannins in a ruminant digestive tract. Insoluble tannin-protein interactions were found at pH 4–5–5–5 for bovine serum albumin and 3–5–5–5 for plant leaf protein. The present study showed that pH alone was not the sole determinant for tannin-protein complex formation, since tannin-protein complexation was found in the pH range 6-0–6-5 when different inorganic ions were added to the solutions. Insoluble complexes were not formed with salivary proteins, although precipitation by tannic acid was achieved at 5°C. This suggests that tannins may form soluble rather than insoluble complexes with salivary proteins. It was concluded that purified F1 leaf protein (the major protei occurring in leaf tissue) ought to be used as the test protein for evaluating tannin-protein interactions for in vitro assay procedures. Using this method it was calculated that 27–43% and 19–40% of available plant protein may interact with condensed tannins from Desmodium intortum and Lotus pedunculatus, respectively.     

Short-term effects of soybean oil supplementation on performance,digestion, and metabolism in dairy cows fed sugarcane-based diets

We aimed to quantify the productive and metabolic responses, and digestive changes in dairy cows fed various concentrations of soybean oil (SBO) in high-concentrate, sugarcane-based diets. Eight rumen-cannulated multiparous Holstein cows in mid lactation (574 ± 19.1 kg of body weight and 122 ± 6.9 d in milk), averaging 22.5 ± 1.22 kg/d of milk were assigned to replicated 4 × 4 Latin squares. The experimental period lasted 21 d as follows: 14 d for adaptation, followed by a sampling period from d 15 to 21. The diets were formulated with increasing concentrations of SBO [% of dry matter (DM)]: control (0%), low (LSBO; 1.57%), medium (MSBO; 4.43%), and high (HSBO; 7.34%). Dry matter intake decreased quadratically in response to SBO addition. The greatest decrease in DM intake was observed in MSBO and HSBO diets. Both milk and energy-corrected milk yield were quadratically affected by the SBO inclusion, with a slight decrease up to MSBO and substantial decrease in the HSBO diet. The milk fat concentration linearly decreased from 3.78% in the control to 3.50% in the HSBO diet. The potentially digestible neutral detergent fiber digestibility in the rumen decreased from 55.7% in the control to 35.2% in the HSBO diet. The fractional rate of digestion of potentially digestible neutral detergent fiber in the rumen decreased linearly from 3.13 to 1.39%/h from the control to HSBO diet. The fractional rate of indigestible neutral detergent fiber passage in the rumen was quadratically affected, with the lowest value (2.25%/h) for the HSBO diet. Rumen pH increased from 6.42 to 6.67, and ammonia nitrogen decreased from 28.1 to 21.4 mg/dL, in the control and HSBO diets, respectively. Rumen volatile fatty acids decreased quadratically, with the greatest decrease observed in MSBO and HSBO diets. Serum concentrations of glucose, fatty acids, and β-hydroxybutyrate were unaffected by SBO inclusion. However, serum concentrations of total cholesterol and high- and low-density lipoproteins linearly increased with increasing concentrations of SBO in the diet. Inclusion of SBO at concentrations from 4.43 to 7.34% of the diet DM decreased DM intake, energy-corrected milk production, fiber digestibility, and rumen fermentation and was thus not recommended. Soybean oil supplementation at 1.57% of the diet DM proved to be a safe concentration for dairy cows fed high-concentrate diets with sugarcane as the sole forage.     

Effect of adding an anaerobic fungal culture isolated from a wild blue bull (Boselophus tragocamelus) to rumen fluid from buffaloes on in vitro fibrolytic enzyme activity,fermentation and degradation of tannins and tannin‐containing Kachnar tree (Bauhinia variegata) leaves and wheat straw

The study investigated the effects of adding an anaerobic fungus (Piromyces sp FNG5; isolated from the faeces of a wild blue bull) to the rumen fluid of buffaloes consuming a basal diet of wheat straw and concentrates on in vitro enzyme activities, fermentation and degradation of tannins and tannin‐rich tree leaves and wheat straw. In experiment 1, strained rumen fluid was incubated for 24 and 48 h, in quadruplicate, with or without fungal culture using condensed tannin‐rich Bauhinia variegata leaves as substrates. In experiment 2, in vitro incubation medium containing wheat straw and different concentrations of added tannic acid (0–1.2 mg mL^?1) were incubated for 48 h, in quadruplicate, with strained buffalo rumen fluid with or without fungal culture. In experiment 3, tolerance of the fungal isolate to tannic acid was tested by estimating fungal growth in pure culture medium containing different concentrations (0–50 g L^?1) of tannic acid. In in vitro studies with Bauhinia variegata tree leaves, addition of the fungal isolate to buffalo strained rumen liquor resulted in significant (P < 0.01) increase in neutral detergent fibre (NDF) digestibility and activities of carboxymethyl cellulase (P < 0.05) and xylanase (P < 0.05) at 24 h fermentation. There was 12.35% increase (P < 0.01) in condensed tannin (CT) degradation on addition of the fungal isolate at 48 h fermentation. In in vitro studies with wheat straw, addition of the fungus caused an increase in apparent digestibility (P < 0.01), true digestibility (P < 0.05), NDF digestibility (P < 0.05), activities of carboxymethyl cellulase (P < 0.001), β‐glucosidase (P < 0.001), xylanase (P < 0.001), acetyl esterase (P < 0.001) and degradation of tannic acid (P < 0.05). Rumen liquor from buffaloes which had never been exposed to tannin‐containing diet had been found to have substantial inherent tannic acid‐degrading ability (degraded 55.3% of added tannic acid within 24 h of fermentation). The fungus could tolerate tannic acid concentration up to 20 g L^?1 in growth medium. The results of this study suggest that introduction of an anaerobic fungal isolate with superior lignocellulolytic activity isolated from the faeces of a wild herbivore may improve fibre digestion from tannin‐containing feeds and degradation of tannins in the rumen of buffaloes. Copyright © 2005 Society of Chemical Industry     

The effects of rumen nitrogen balance on nutrient intake,nitrogen partitioning,and microbial protein synthesis in lactating dairy cows offered different dietary protein sources

The aim was to study the effects of rumen N balance (RNB), dietary protein source, and their interaction on feed intake, N partitioning, and rumen microbial crude protein (MCP) synthesis in lactating dairy cows. Twenty-four lactating Holstein cows were included in a replicated 4 × 4 Latin square experimental design comprising four 20-d periods, each with 12 d of adaptation to the experimental diets and 8 d of sampling. The dietary treatments followed a 2 × 2 factorial arrangement (i.e., 4 treatments) with 2 main protein sources [faba bean grain (FB) and SoyPass (SP; Beweka Kraftfutterwerk GmbH, Heilbronn, Germany)] offered at 2 dietary RNB levels each [0 g/kg of dry matter, DM (RNB0) and ?3.2 g/kg of DM (RNB?)]. The RNB was calculated as the difference between dietary crude protein (CP) intake and the rumen outflow of undegraded feed CP and MCP and divided by 6.25. Composition of concentrate mixtures was adjusted to create diets with desired RNB levels. Each of these protein sources supplied ≥35% of total dietary CP. Both diets for each protein source were isoenergetic but differed in CP concentrations. The DM intake (kg/d) was lower for RNB? than for RNB0 in diets containing FB, whereas no differences were seen between the RNB levels for SP diets. The RNB? decreased N intake and urinary N excretion but increased milk N use efficiency in both FB and SP diets, with greater differences between the RNB levels for FB diets than for SP diets. Similarly, duodenal MCP synthesis (g/kg of digestible organic matter intake) estimated from purine derivatives in the urine was lower for RNB? than for RNB0 in FB diets but similar between the RNB levels in diets containing SP. Low RNB of approximately ?65 g/d (approximately ?3.2 g/kg of DM) in diets reduced feed intake, N balance, and performance in high-yielding dairy cows with possibly more pronounced effects in diets containing rapidly degradable protein sources.     

In vitro evaluation of digestive enzyme inhibition and antioxidant effects of naked oat phenolic acid compound (OPC)

This research was focused on digestive enzyme inhibition and antioxidant properties of naked oat phenolic acid compound (OPC). Free and bound phenolic acid were separated from ethyl acetate fraction, n-butanol fraction and aqueous fraction. The interactions between OPC and main digestive enzymes (α-amylase, α-glucosidase, pepsin and trypsin) were studied. It was shown that the semi-purified bound phenolic acid (semi-purified by AB-8 column) has a competitive alpha-glucosidase inhibitor, while OPC of the organic extract fraction exhibited the characteristics of a mixed inhibitor. Bound phenolic-n-butanol fraction (IC₅₀ = 98.39 ± 0.89 µg mL⁻¹) had the strongest ability to scavenge the 1,1-diphenyl-2-picrylhydrazyl (DPPH). Additionally, the starch hydrolysis degree of n-butanol extraction naked oat phenolic compound was significantly lower than other fractions in vitro. The integrated results suggested that OPC could be considered as potential healthy factor to control postprandial blood glucose, and the mechanism maybe via anti-digestion, antioxidation and interaction with diabetes-related starch.     

Difference in the nature of tannins on in vitro ruminal methane and volatile fatty acid production and on methanogenic archaea and protozoal populations

Six plant sources of hydrolyzable tannins (HT) or HT and condensed tannins (CT; designated as HT1, HT2, HT3, HT + CT1, HT + CT2, and HT + CT3) were evaluated to determine their effects in vitro on CH₄ production and on ruminal archaeal and protozoa populations, and to assess potential differences in biological activities between sources containing HT only or HT and CT. Samples HT1, HT2, and HT3 contained only HT, whereas samples HT + CT1, HT + CT2, and HT + CT3 contained HT and CT. In experiment 1, in vitro incubations with samples containing HT or HT + CT resulted in a decrease in CH₄ production of 0.6 and 5.5%, respectively, compared with that produced by incubations containing the added tannin binder polyethylene glycol-6000. Tannin also suppressed the population of methanogenic archaea in all incubations except those with HT2, with an average decrease of 11.6% in HT incubations (15.8, 7.09, and 12.0 in HT1, HT2, and HT3) and 28.6% in incubations containing HT + CT (35.0, 40.1, and 10.8 in HT + CT1, HT + CT2, and HT + CT3) when compared with incubations containing added polyethylene glycol-6000. The mean decrease in protozoal counts was 12.3% in HT and 36.2% in HT + CT incubations. Tannins increased in vitro pH, reduced total VFA concentrations, increased propionate concentrations, and decreased concentrations of iso-acids. In experiment 2, when a basal diet was incubated with graded levels of HT + CT1, HT + CT2, and HT + CT3, the total gas and CH₄ production and archaeal and protozoal populations decreased as the concentration of tannins increased. Our results confirm that tannins suppress methanogenesis by reducing methanogenic populations in the rumen either directly or by reducing the protozoal population, thereby reducing methanogens symbiotically associated with the protozoal population. In addition, tannin sources containing both HT and CT were more potent in suppressing methanogenesis than those containing only HT.     

苦瓜皂苷提取物微胶囊化缓释性能及体外对糖苷酶活性影响研究

以壳聚糖和海藻酸钠为壁材,采用锐孔凝固浴法制备苦瓜皂苷提取物微胶囊,并对其进行了缓释性能及在模拟胃肠酸碱环境中对α-葡萄糖苷酶和α-淀粉酶活性影响的研究。结果显示,微胶囊在体外模拟胃肠液中具有良好的缓释性能,在模拟胃液中苦瓜皂苷1h释放率为31.19%,2h累积释放率为52.14%,转至肠液中10h累积释放率达到81.20%;在模拟胃肠液中微胶囊对α-葡萄糖苷酶和α-淀粉酶均有一定的抑制作用,抑制率随着苦瓜皂苷释放量的增加有上升趋势。表明苦瓜皂苷经微胶囊化后能够保护其活性,缓解副作用和延长对糖苷酶的作用时间。      

Effects of variation in soluble carbohydrate intake on sulfur dynamics in the sheep rumen

Two groups of experiments were conducted (named the 15% starch and 30% starch intake experiments) to examine the effects of dietary energy levels on the synthesis of ruminal microbial protein and the absorption of sulfur from the rumen. The level of energy intake affected the absorption rate of sulfide from the rumen, the net microbial protein flow rate, the amount of microbial protein synthesized per kg organic matter ‘truly digested’ in the rumen and the daily flow of recycled sulfur. For the 15% starch and 30% starch intake experiments the absorption of sulfur from the rumen was 1.207 and 1.278 g day^?1 absorbed from 2.298 and 2.212 g intake, respectively, the flow of microbial protein was 80.2 and 92.1 g day^?1, respectively, and the estimated recycled sulfur was 0middot;357 and 0.670 g day^?1, respectively. It was estimated that 18.1 and 20.1 g of microbial protein was synthesized per 100 g of organic matter ‘truly digested’ in the rumen for 15% starch and 30% starch diets, respectively.     

Comparison of grass and legume silages for milk production. 2. In vivo and in sacco evaluations of rumen function

Two experiments were conducted to investigate the basis for higher voluntary intakes and increased alpha-linolenic acid content in milk from cows offered clover silages. Six cows with rumen and duodenal cannulae were used in a four-period changeover-design experiment. Cows received 8 kg/d of dairy concentrate and had ad libitum access to one of six silage treatments: grass, red clover, white clover, alfalfa, and 50/50 (dry matter basis) mixtures of grass with red clover or white clover. The rumen fermentability of grass, red clover, white clover, and grass/red clover silages was also evaluated in a nylon bag study. Legume silages led to increased dry matter intake and milk production in comparison with grass silage. There was no significant effect of legume silages on rumen pH and volatile fatty acid concentrations, but a significant increase in rumen ammonia concentration with the legume silages, reflecting their higher protein content. The inclusion of white clover or alfalfa silage, but not red clover silage, in diets led to an increase in molar proportions of isobutyric, iso-valeric, and n-valeric acids in comparison with diets based on grass silage. Rumen fill was significantly lower, and rumen passage rates were significantly higher for cows offered alfalfa or white clover silages. However, the markedly different particle size distribution of rumen contents with these feeds suggests very different mechanisms for the high intake characteristics: high rates of particle breakdown and passage with alfalfa, and high rates of fermentation and passage with white clover. Microbial energetic efficiency (grams microbial N per kilogram organic matter apparently digested in the rumen) was highest for cows offered alfalfa silage, intermediate for clover silage, and lowest for cows offered grass silage. These differences reflect the higher rumen outflow rates for legume silages in comparison with grass silage. However, the effect of these differences on N-use efficiency (feed to milk) was probably quite small in comparison with effects of N intake. Although the biohydrogenation of alpha-linolenic acid was still high for red clover silage (86.1% compared with 94.3% for grass silage), there was a 240% increase in the proportion of alpha-linolenic acid passing through the rumen. This explains the increased recovery of alpha-linolenic acid from feed into milk with diets based on red clover silage.     

Effects of a commercial fermentation byproduct or urea on milk production,rumen metabolism,and omasal flow of nutrients in lactating dairy cattle

The objective of this study was to evaluate the effects of a fermentation byproduct on rumen fermentation and microbial yield in high producing lactating dairy cattle. Eight ruminally cannulated multiparous Holstein cows averaging (mean ± standard deviation) 60 ± 10 d in milk and 637 ± 38 kg of body weight were assigned to 1 of 2 treatment sequences in a switchback design. Treatment diets contained (dry matter basis) 44% corn silage, 13% alfalfa silage, 12% ground corn, and 31% premix containing either a control mix of urea and wheat middlings (CON) or a commercial fermentation byproduct meal (Fermenten, Arm and Hammer Animal Nutrition, Princeton, NJ) at 3% diet inclusion rate (EXP). Diets were formulated to be isonitrogenous and isocaloric, with similar levels of neutral detergent fiber and starch. The trial consisted of three 28-d experimental periods, where each period consisted of 21 d of diet adaptation and 7 d of data and sample collection. Omasal nutrient flows were determined using a triple-marker technique and double-labeled ¹⁵N¹⁵N-urea. The EXP diet provided 18 g/d more nonammonia N versus the CON diet, representing 3.0% of total N intake. Energy-corrected milk yield (41.7 and 43.1 kg/d for CON and EXP, respectively), milk fat, and protein yield and content did not differ between treatments. Total dry matter intake was similar between treatments (25.5 and 26.4 kg/d for CON and EXP, respectively). Ammonia N concentration and pool size in the rumen was greater in cows fed the EXP diet. No differences were observed in rumen or total-tract dry matter, organic matter, or neutral detergent fiber digestibility. Ruminal degradation of feed N was 15% lower in cows fed EXP diets, resulting in differences in omasal N flows. Results demonstrated the fermentation byproduct meal had a sparing effect on degradable feed protein, but did not increase microbial N flow from the rumen.     

Degradation of condensed tannins by rumen microbes exposed to quebracho tannins (QT) in rumen simulation technique (RUSITEC) and effects of QT on fermentative processes in the RUSITEC

In an earlier study it was observed that the rumen microbes from cattle which had never consumed tannin-containing diets do not have enzymes for degrading condensed tannins. In this study, rumen microbes were exposed to small amounts of quebracho tannins for 8 days using a rumen simulation technique (RUSITEC). The levels maintained in the RUSITEC were 0.1, 0.2 and 0.4 mg of the spray-dried quebracho powder (SDQT) per ml of the medium. After 8 days of tannin exposure, the liquor containing ‘exposed/adapted’ microbes from RUSITEC was incubated for 40 h without and with purified tannins of quebracho and Dichostachys cinerea leaves in in vitro gas method. There was no degradation of condensed tannins. The enzymes for degradation of condensed tannins were not induced in rumen microbes by exposure to different concentrations of tannins for 8 days in the RUSITEC. In the RUSITEC, SDQT significantly reduced the number of total protozoa, entodiniomorphs and holotrichs; effect was higher on holotrichs. There was no significant change in the levels of short chain fatty acid but the molar proportion of propionate was significantly higher and of butyrate significantly lower at 0.4 mg SDQT ml^?1. Significantly lower levels of ammonia in the medium was also observed on injection of tannins. Microbial mass production, calculated using ¹⁵N incorporation, was similar at 0.1 and 0.2 mg SDQT ml^?1 but significantly lower (13%) at 0.4 mg SDQT ml^?1. The dry matter digestibility of the feed (80% hay and 20% barley) was not significantly affected by SDQT.     

Condensed tannins from Lotus corniculatus and Lotus pedunculatus exert different effects on the in vitro rumen degradation of ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco) protein

Condensed tannins (CT) or proanthocyanidins (PA), which occur in a restricted range of forages, have the ability to interact with proteins and enzymes and can influence the digestion of plant protein in the rumen. We compared the effects of CT extracts from Lotus corniculatus and pedunculatus on degradation of the principal leaf protein, ribulose‐1,5‐bisphosphate carboxylase/oxygenase (Rubisco), by rumen microorganisms. Total soluble leaf protein extracted from white clover (Trifolium repens ) was incubated with fresh rumen fluid from sheep and a range of concentrations of each CT extract. The rate of degradation of the large (LSU) and small subunit (SSU) of Rubisco was quantified by fractionating the proteins in samples taken from in vitro rumen incubations using sodium dodecyl sulphate–polyacrylamide gel electrophoresis (SDS‐PAGE) and imaging densitometry. To deduce the effects of the CT extracts, experiments were performed in the presence (CT inactive) and absence (CT active) of polyethylene glycol (PEG; MW 3350). The two CT extracts differed markedly in their effects on the degradation of the LSU and SSU of Rubisco. At concentrations of 0.89 and 1.79 mg CT mg ⁻¹ total soluble leaf protein, the CT extract from L pedunculatus was more effective at preventing the degradation of the LSU and SSU by rumen microorganisms than the CT extract from L corniculatus. At a concentration of 1.79 mg CT mg ⁻¹ total soluble leaf protein, the CT extracts from L corniculatus and pedunculatus prevented about 0.75 and 0.83 of the LSU and about 0.69 and 0.86 of the SSU, respectively, from being degraded. Addition of PEG removed the inhibition and almost complete degradation of these proteins occurred, as was the case in incubations without CT extracts. The results of this study suggest that the concentration of CT in the diet and the chemical structure which affects the activity of the CT needs to be considered when assessing the effects of CT on protein metabolism in ruminants. © 1999 Society of Chemical Industry