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Binding affinity of hydrolyzable tannins to parotid saliva and to proline-rich proteins derived from it.

Proline-rich proteins (PRP) in human parotid saliva have a high affinity for dietary polyphenolic compounds (tannins), forming stable complexes that may modulate the biological and nutritional properties of the tannin. The formation of such complexes may also have an important role in the modulation or promotion of the sensation of oral astringency perceived when tannin-rich foods and beverages are consumed. The major classes of PRP (acidic, basic, and glycosylated) have been isolated from human saliva, and the relative binding affinities of a series of hydrolyzable tannins, which are found in a number of plant-derived foods and beverages, to these PRP classes have been determined using a competition assay. All of the classes of PRP have a high capacity for hydrolyzable tannins. Within the narrow range of binding affinities exhibited, structure/binding relationships with the levels of tannin galloylation, hexahydroxydiphenoyl esterification, and degree of polymerization were identified. No individual class of human salivary PRP appears to have an exclusive affinity for a particular type of hydrolyzable tannin.

Electrophoresis, Polyacrylamide Gel↗

Insoluble condensed tannins of canola/rapeseed.

The contents of soluble, SDS-extractable, and insoluble condensed tannins were determined in canola/rapeseed hulls from several varieties by utilizing the proanthocyanidin assay. The total amount of tannins in rapeseed/canola hulls ranged from 1913 to 6213 mg per 100 g of oil-free hulls. Insoluble tannins predominated in canola/rapeseed hulls and comprised from 70 to 95.8% of total tannins present. The amounts of SDS-extractable tannins were comparable to those of soluble tannins but constituted only 4.7-14. 1% of insoluble tannins present.

Anthocyanins↗

The role of condensed tannins in the nutritional value of Lotus pedunculatus for sheep. 1. Voluntary intake.

Voluntary intake was determined with vegetative Lotus pedunculatus cut and fed fresh to growing sheep of 42-46 kg live weight. Effects attributable to condensed tannins were assessed by growing the plant under high and low levels of soil fertility, inducing low and high concentrations of tannin (Expt 1), or by binding the tannins through spraying the herbage with polyethylene glycol (molecular weight 3350, PEG; Expt 2). Primary-growth lotus was used in Expt 1 and secondary-growth lotus in Expt 2. Concentrations of total and free condensed tannin were determined in fresh lotus, free tanning being defined as that not bound by mascerates of the plant. In Expt 1 the herbages fed contained respectively 46 and 106 g total condensed tannin/kg dry matter (DM) and 3 and 14 g free condensed tannin/kg DM. Mean metabolizable energy (ME) intakes were 0.89 and 0.77 MJ/kg live weight0 .75 per d (P less than 0.05) respectively. The lotus used in Expt 2 contained 63 and 5 g total reactive condensed tannin and free condensed tannin/kg DM respectively. After spraying with PEG at 2.4 g/g total condensed tannin, these values were reduced to 7 and 0.5 g/kg DM respectively. PEG addition increased apparent digestibility (proportion of each nutrient ingested) of cellulose, hemicellulose and nitrogen by 0.05, 0.08 and 0.26, and increased ME intake from 0.48 to 0.69 MJ/kg live weight0 .75 per d. It was concluded that high concentrations of condensed tannins depressed ME intake, due to depressions in both the voluntary intake and digestion of organic matter.(ABSTRACT TRUNCATED AT 250 WORDS)

Animal Feed↗

Rumen fermentation and nitrogen balance of lambs fed diets containing plant extracts rich in tannins and saponins, and associated emissions of nitrogen and methane.

Tannins were added to experimental diets at levels of 1 and 2 g/kg DM (hydrolysable tannins; Castanea sativa wood extract) and saponins at 2 and 30 mg/kg DM (sarsaponin; Yucca schidigera extract). These levels were far below thresholds expected to be adverse in ruminants. Effects were measured in lambs by comparison with unsupplemented control diets calculated to be either deficient (10%) or adequate in protein. The diets consisted of hay, concentrate (1:1) and extra wheat starch with increasing body weight. Ruminal pH, VFA concentration, protozoa count and apparent digestibilities of organic matter and fibre did not differ among treatments. The low tannin dose significantly decreased bacteria count compared to the high saponin dose. Saponin supplementation and the high tannin dose showed some potential to reduce ruminal ammonia concentration. This was associated with weak trends towards lower urine N excretion (only tannins) and ammonia emission from manure. Methane release was increased by the low tannin dose compared to the unsupplemented control. Diet effects on heat production were not systematic. In conclusion, the extracts rich in tannins or saponins gave only slight indications for either increased body nitrogen retention or reduced nitrogen emission. However, effects might have been larger with more pronounced dietary protein deficit.

Animal Feed↗

Effect of tannins on galliform cecal partitioning.

Winter diets of ruffed grouse (Bonasa umbellus) and other galliformes are high in tannins. We fed quebracho, a condensed tannin, to ruffed grouse and found no effect on dry matter intake or body mass at levels up to 6% of the diet. However, a high-fiber diet with 8% quebracho resulted in reduced dry matter intake and body mass loss. Grouse could not tolerate a diet with 8% tannic acid, a hydrolyzable tannin, which caused a large reduction in dry matter intake and body mass. Northern bobwhite (Colinus virginianus) and ruffed grouse responded to dietary quebracho tannin by increasing the proportion of digesta that was excreted from the ceca. In the northern bobwhite, 59%-76% of the tannin recovered from the feces was in cecal feces. There was no difference in average passage rates of liquid and fiber digesta, although variation was high in tannin-fed birds. The role of the ceca in handling tannin requires further investigation.

Animals↗

Dietary tannins from cowpeas and tea transiently alter apparent calcium absorption but not absorption and utilization of protein in rats.

Tannins reportedly alter absorption and utilization of protein and minerals. The present study investigated the effect of tannins extracted from 'Mississippi Silver' cowpeas and black tea when incorporated into nutritionally balanced diets. Condensed tannins were incorporated into the diet of weanling male Sprague-Dawley rats at 0.0, 0.0057, 0.0171 and 0.057 g/100 g diet for 28 d. Ingestion of tannin from cowpeas or tea did not change significantly growth rate, protein efficiency ratio, apparent protein digestibility, nitrogen excretion, relative liver weight, or nitrogen concentration of liver. During d 11-18, apparent calcium absorption was lower in rats fed the medium and high levels of cowpea tannin and in those fed all levels of tea tannin compared with the control group. By wk 4, no differences were observed in apparent calcium absorption among treatment groups. Apparent magnesium absorption was not affected by dietary treatment, nor was femur content of calcium or magnesium. We conclude that at the levels of condensed tannins fed, a short-term reduction of apparent calcium absorption occurred; however, by wk 4 calcium absorption was comparable to that of the control group. The acute change that occurred in apparent calcium absorption did not influence bone calcium content.

Absorption↗

Antioxidant status of faeces of captive black rhinoceros (Diceros bicornis) in relation to dietary tannin supplementation.

In context with the frequent observations of excessive iron (Fe) storage in captive black rhinoceroses (Diceros bicornis), it has been suggested that both an excessive dietary Fe content and a lack of dietary Fe-chelating substances, such as tannins, is the underlying cause. Therefore, studies on the effects of tannin supplementation to captive diet are warranted. Six captive rhinoceroses were fed their normal zoo diet (N), and a similar diet supplemented with either tannic acid (T, hydrolysable tannin) or quebracho (Q, condensed tannins), and the total antioxidant capacity (TAC) was measured as mmol Trolox equivalents per kg fresh faeces. The TAC values on diets N (1.24 +/- 0.39 mmol/kg fresh faeces) and T (1.34 +/- 0.33 mmol/kg fresh faeces) were similar, but significantly higher on diet Q (2.32 +/- 0.61 mmol/kg fresh faeces). In contrast to expectations, faecal TAC increased with increasing faecal Fe, possibly as a result of the fact that the faecal Fe content was positively correlated to the proportion of concentrate feeds in the diet, which also contain antioxidants, such as vitamin E, in addition to Fe. Increased antioxidant status caused by the use of tannin substances could have a beneficial effect on animal health, but if tannins should be incorporated in designed diets, other tannin sources, such as grape pomace should be tested.

Animals↗

Isolation and characterization of proteolytic ruminal bacteria from sheep and goats fed the tannin-containing shrub legume Calliandra calothyrsus.

Tannins in forages complex with protein and reduce the availability of nitrogen to ruminants. Ruminal bacteria that ferment protein or peptides in the presence of tannins may benefit digestion of these diets. Bacteria from the rumina of sheep and goats fed Calliandra calothyrsus (3.6% N and 6% condensed tannin) were isolated on proteinaceous agar medium overlaid with either condensed (calliandra tannin) or hydrolyzable (tannic acid) tannin. Fifteen genotypes were identified, based on 16S ribosomal DNA-restriction fragment length polymorphism analysis, and all were proteolytic and fermented peptides to ammonia. Ten of the isolates grew to high optical density (OD) on carbohydrates (glucose, cellobiose, xylose, xylan, starch, and maltose), while the other isolates did not utilize or had low growth on these substrates. In pure culture, representative isolates were unable to ferment protein that was present in calliandra or had been complexed with tannin. One isolate, Lp1284, had high protease activity (80 U), a high specific growth rate (0.28), and a high rate of ammonia production (734 nmol/min/ml/OD unit) on Casamino Acids and Trypticase Peptone. Phylogenetic analysis of the 16S ribosomal DNA sequence showed that Lp1284 was related (97. 6%) to Clostridium botulinum NCTC 7273. Purified plant protein and casein also supported growth of Lp1284 and were fermented to ammonia. This is the first report of a proteolytic, ammonia-hyperproducing bacterium from the rumen. In conclusion, a diverse group of proteolytic and peptidolytic bacteria were present in the rumen, but the isolates could not digest protein that was complexed with condensed tannin.

Animal Feed↗

Cotton bract tannin: a novel human T-lymphocyte mitogen and a possible causative agent of byssinosis.

Cotton bract tannin, a major organic component of cotton dust, was tested for mitogenic activity upon human T lymphocytes. Tannin caused polyclonal activation of human T lymphocytes in vitro. The pattern of T-cell response to tannin was similar to the pattern seen in lymphocytes stimulated with the well-known plant lectin T-cell mitogens, PHA and ConA. The response to tannin was shown to be dependent upon tannin dose and presence of monocytes. The maximum response occurred after 3-4 days in culture with the magnitude comparable to the one achieved by ConA. Lipopolysaccharide from Enterobacter agglomerans, a major contaminant of cotton dust, was shown not to be a mediator of tannin mitogenicity in vitro. A role for tannin as a polyclonal cell activator and, therefore, a possible etiologic agent of byssinosis is suggested by these observations.

Antigen-Presenting Cells↗

Tannin-mediated secretion of a neutrophil chemotactic factor from alveolar macrophages. Potential contribution to the acute pulmonary inflammatory reaction associated with byssinosis.

Inhalation of cotton mill dust or condensed tannin, a major botanical component of cotton mill dust, induces an acute pulmonary inflammatory response characterized by a rapid influx of neutrophils into the airways. The development of neutrophil alveolitis caused by tannin inhalation is accompanied by the accumulation of low molecular weight neutrophil chemotactic factor (NCF) in the airways. To determine if the alveolar macrophage is the source of this NCF, the ability of tannin to induce the secretion of NCF from rabbit alveolar macrophages was examined in vitro. Tannin did promote the secretion of NCF from alveolar macrophages in a time- and dose-dependent manner. Secretion began immediately after challenge and was maximal after 4 to 5 h. Maximal secretion occurred at a tannin concentration of 50 micrograms/ml. Comparison with the dose response for NCF secretion by cotton dust extract indicated that tannin was the major component in the dust responsible for NCF secretion from alveolar macrophages in the time period examined. The NCF had an apparent molecular weight of greater than 800 as determined by gel chromatography. The NCF could be extracted into organic solvents, suggesting it was a lipid. Its secretion, however, could not be prevented by treatment of the macrophages with the 5-lipoxygenase inhibitor, nordihydroguaretic acid, demonstrating that the NCF was not leukotriene B4. These data indicate that the action of tannin on resident alveolar macrophages results in the secretion of a NCF that may be responsible for the acute neutrophil alveolitis associated with inhalation of cotton dust.

Animals↗

Signal transducing mechanisms in human platelets stimulated by cotton bract tannin.

Cotton bract tannin is a potent stimulus for platelet aggregation and secretion. Tannin has been shown to stimulate the phosphorylation of two 19-kD and 47-kD cytosolic proteins in platelets, but earlier steps in signal transducing mechanisms of platelet activation are unknown. In this study, measurements of 32P-labeled phospholipids, 14C-labeled arachidonic acid, and levels of intracellular free calcium (Ca2+) were performed before and after the addition of thrombin (1 U/ml) or various concentrations of tannin to human platelets. The results showed that tannin induced a dose-dependent synthesis of phosphatidic acid, an early and transient hydrolysis of phosphatidylinositol monophosphate and bisphosphate, a transient synthesis of diacylglycerol, and a release of arachidonic acid metabolites. The kinetics of phosphatidic acid, diacylglycerol, and arachidonic acid metabolite synthesis were similar after platelet stimulation by tannin (75 micrograms/ml) or thrombin. Tannin also induced a reversible rise of intracellular Ca2+ due to a mobilization of the internal stores and an influx of extracellular Ca2+. These results suggest that cotton bract tannin, as thrombin, activates human platelets by phospholipase C and A2 activations, release of diacylglycerol, and mobilization of intracellular free Ca2+.

Arachidonic Acids↗

Effects of green tea tannin on cisplatin-induced nephropathy in LLC-PK1 cells and rats.

A study was conducted to clarify whether green tea tannin ameliorated cisplatin-induced renal injury in terms of lactate dehydrogenase and malondialdehyde leakage from a renal epithelial cell line, swine-derived LLC-PK1 cells in culture. Green tea tannin was shown to suppress the cytotoxicity of cisplatin, the suppressive effect increasing with the dose of green tea tannin. The effect of cisplatin was then investigated in rats given green tea tannin for 40 days before cisplatin administration and in control rats given no green tea tannin. In control rats, blood, urinary and renal parameters and the activities of antioxidative enzymes in renal tissue deviated from the normal range, indicating dysfunction of the kidneys. In contrast, rats given green tea tannin showed decreased blood levels of urea nitrogen and creatinine, and decreased urinary levels of protein and glucose, reflecting less damage to the kidney. In this group, the activity of catalase in the renal tissue was increased, while the level of malondialdehyde was decreased, suggesting the involvement of radicals in the normalizing of kidney function. Based on the evidence available it appeared that green tea tannin eliminated oxidative stress and was beneficial to renal function.

Animals↗

Effect of chestnut tannin on fermentation quality, proteolysis, and protein rumen degradability of alfalfa silage.

Two experiments were conducted on alfalfa to investigate the effects of the addition of commercial chestnut hydrolyzable tannin at ensiling on 1) silage fermentation quality in lab-scale silos and protein degradation in the rumen, and 2) silage fermentation quality and proteolysis in bale silages. Wilted alfalfa was prepared with 4 tannin levels (0, 2, 4, and 6% on a dry matter (DM) basis; T0, T1, T2, T3, respectively) and ensiled in lab-scale silos. Silages (33% DM) were analyzed for fermentation quality, protein rumen degradability in situ, and organic matter digestibility in vitro through gas production after 120 d of conservation. Wilted alfalfa containing 0 and 4% tannin (T0 and T2) was harvested at 40% DM (wilting level I) and 53% DM (wilting level II) for bale (600 mm diameter) silage. Silages were analyzed for fermentation quality after 78 d of conservation. All the silages were well fermented with no butyric acid. Lab-scale silages showed reductions in ammonia, nonprotein nitrogen (NPN) and DM losses in T2 and T3 treatments, while the fermentation acid profiles were unaffected. In experiment 1, the untreated silage (T0) had the highest protein degradability after being incubated in the rumen. The addition of tannin reduced crude protein ruminal disappearance in a dose-dependent manner. However, the tannin reduced the organic matter digestibility by 5.1% for all of the tannin addition levels. The tannin positively affected the silage quality in the round bale silages, in particular reducing ammonia and NPN in the lowest wilting level. In both experiments, T2 treatment reduced proteolysis without any influence of DM on the binding reaction and reduced the NPN by 15% in comparison to the control.

Ammonia↗

Improvement of the nutritive value of high tannin sorghums for broiler chickens by high moisture storage (reconstitution).

Grains from three high and one low tannin sorghum cultivars were treated as follows: 1) untreated, no water was added, the dry grain had an average moisture content of 8% and 2) the moisture content of the grain was raised by adding 25% (w/w) distilled water to the grain. The treated grain was then stored for 10 or 20 days with a 2% level of acetic-propionic acid mixture (60/40, v/v) added to deter fungal growth. This treated grain was used in diets during a 3-week feeding trial with broiler chickens. High moisture storage (reconstitution) of high tannin sorghums decreased their extractable tannin content. Chickens fed diets containing these treated sorghums had improved weight grains (23 to 83 g) and feed efficiencies compared to birds fed diets containing the untreated grain. Treatment of low tannin sorghum did not improve weight gains. Reconstitution of high but not low tannin sorghums improved protein digestibility (6 to 16%) and dietary metabolizable energy (.1 to .3 kcal/g) for broiler chickens. There was a highly significant cultivar effect on leg scores, but the correlation between dietary tannin content and leg scores was low (r = .10), suggesting that tannin content was not a major factor in leg abnormality incidences shown. The improvement in weight grains, feed efficiencies, and protein digestibilities at 10 and 20 days storage periods was not different (P greater than .05), indicating that the benefits of high moisture storage were already obtained by 10 days.

Animals↗

Detoxification of high tannin sorghum grains.

One in vitro and two in vivo experiments were conducted to determine appropriate methodology for and effects of detoxifying Darset, Redlan, and one commercial hybrid brown sorghum grain variety in threonine-deficient and nutritionally complete poultry rations. The detoxification procedure, which involves adding water to sorghum grain to bring dry matter to 70% and subsequent anaerobic incubation at 32 C, removes up to 100% of the chemically detectable tannin. Rate of tannin elimination was dependent upon sorghum grain variety with commercial brown sorghum requiring approximately 3 days longer than the Darset for tannin elimination. Detoxification of high tannin sorghum grains improved (P less than .05) growth rate and feed efficiency dramatically in threonine-deficient, crystalline amino acid-fortified rations; the same process applied to low tannin sorghum grains was without benefit. The desirable effects of this type of grain processing appear to be due to the reduced grain tannin content. Including detoxified sorghum grains in a practical-type broiler ration failed to elicit a weight gain response although feed efficiency was improved by 13% over untreated sorghum and 3% over corn grain. Lack of a gain response was likely due to the excess protein included in the basal diet. This type of grain processing offers an alternative method of reducing sorghum grain tannin content and is likely of greatest value in rations containing marginal indispensable amino acid and protein levels.

Amino Acids↗

Effect of polyethylene glycol on in vitro degradability of nitrogen and microbial protein synthesis from tannin-rich browse and herbaceous legumes.

Determination of microbial degradability of N is important in formulating a sound supplementation strategy for efficient utilisation of basal as well as supplementary diet components. In vitro degradability of N (IVDN) from tannin-containing browses (Acacia cyanophylla, Acacia albida, Acioa barteri and Quercus ilex) and two herbaceous legumes (Desmodium intortum and Desmodium uncinatum) was determined using the in vitro gas-production method coupled with NH3-N measurement in the presence and absence of a tannin-binding agent (polyethylene glycol (PEG), molecular mass 6000). Addition of PEG to tannin-containing feeds significantly (P < 0.05) increased in vitro gas and short-chain fatty acid (SCFA) production, and IVDN. The use of PEG as a tannin-binding agent increased IVDN from 28 to 59, 32 to 72, 19 to 40, 32 to 73, 40 to 80, and 26 to 77% in A. cyanophylla, A. albida, A. barteri, D. intortum, D. uncinatum and Q. ilex respectively. There was significant correlation between total phenolic compounds (total phenol, TP; total tannin, TT) in leguminous forages and percentage increase in IVDN on addition of PEG (P < 0.05; R2 0.70 and 0.82 for TP and TT respectively). The difference in IVDN observed in the absence and presence of PEG indicates the amount of protein protected from degradation in the rumen by tannins. When measured after 24 h incubation, tannin-containing feeds incubated in absence of PEG resulted in higher microbial protein synthesis than in the presence of PEG. Addition of PEG significantly (P < 0.05) reduced the efficiency of microbial protein synthesis expressed as mumol purine/mmol SCFA.

Animals↗

Rapid determination of tannins in tanning baths by adaptation of BSA method.

A rapid and reproducible method for the determination of tannins in vegetable tanning baths is proposed as a modification of the BSA method for grain tannins existing in literature. The protein BSA was used instead of leather powder employed in the Filter Method, which is adopted in Italy and various others countries of Central Europe. In this rapid method the tannin contents is determined by means a spectrophotometric reading and not by means a gravimetric analysis of the Filter Method. The BSA method, which belongs to mixed methods (which use both precipitation and complexation of tannins), consists of selective precipitation of tannin from a solution containing also non tannins by BSA, the dissolution of precipitate and the quantification of free tannin amount by its complexation with Fe(III) in hydrochloric solutions. The absorbance values, read at 522 nm, have been expressed in terms of tannic acid concentration by using a calibration curve made with standard solutions of tannic acid; these have been correlated with the results obtained by using the Filter Method.

Absorption↗

[Determination of tannin in cotton plant].

Three methods were used to determine the tannins content in cotton plant tssues. The tannin content in different organs and terminal leaves of four cotton varieties at different growth stages determined by the Folin's assay was nearly two folds as much as that determined by acid butanol assay, indicating that the Folin's assay only offers the total phenol content, but is not suitable for determining the content of condensed tannin in cotton. There was no significant difference between the results of vanillin assay and acid butanol assay, and hence, it is feasible to use vanillin assay to test the tannin content in cotton. Determinations at boll stage showed that the content of condensed tannin in calyx, boll shell and leaf was rather high, generally up to 5-10%, while that in petal, stigma, ovary and inner boll was lower (about 2%). The condensed tannin content in terminal leaf continually increased from seedling(< 1%) to boll stage (10% or so), indicating that the increased tannin content is closely related to the maturity, decrepitude and lignification of cotton tissues.

Gossypium↗