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Activities of enzymes of the pancreas, and the lumen and mucosa of the small intestine in growing broiler cockerels fed on tannin-containing diets.

Diets containing vegetable tannins, predominantly hydrolysable gallotannins, at levels of 13.5, 25 and 50 g/kg were fed to growing broiler cockerels to examine their effect on enzymes in the pancreas, the intestinal lumen and the intestinal mucosa. Pancreas weight per unit live weight showed a significant (P less than 0.05) increase with increasing level of dietary tannin while that of the liver remained unaffected. Trypsin (EC 3.4.21.4) and alpha-amylase (EC 3.2.1.1) activities in the pancreas of birds fed at the highest level of tannins were more than double those from birds fed on a tannin-free control diet. In the intestinal lumen inhibition of trypsin activity increased with increasing level of dietary tannin; alpha-amylase activity was inhibited at intermediate tannin levels but was restored at the highest level. Dipeptidase (EC 3.4.13.11) and sucrose alpha-glucosidase (disaccharidase) (EC 3.2.1.48) in the intestinal mucosa were both inhibited by tannins. Growth of the birds and digestibility of nitrogen were adversely affected by the tannin-containing diets.

Animal Feed↗

Effect of condensed tannins in hulls of faba beans (Vicia faba L.) on the activities of trypsin (EC 2.4.21.4) and chymotrypsin (EC 2.4.21.1) in digesta collected from the small intestine of pigs.

The effects of condensed tannins in hulls of faba beans (Vicia faba L.) on the activities of trypsin (EC 3.4.21.4) and chymotrypsin (EC 3.4.21.1) in digesta obtained from the small intestine of pigs were studied. Using four castrated male pigs (mean body weight 83 kg) fitted with both a simple T-cannula in the duodenum and a post-valvular T-cannula at the terminal ileum, two experimental diets were tested in a Latin square design. The low-tannin diet (LT) contained 200 g faba bean hulls (cv. Blandine)/kg with a low content of condensed tannins (< 0.1% catechin equivalents). The high-tannin diet (HT) contained 200 g faba bean hulls (cv. Alfred)/kg with a content of condensed tannins of 3.5% catechin equivalents. Spot samples of fresh duodenal digesta were taken daily at fifteen time points between 08.00 and 20.00 hours on four consecutive days. Ileal digesta were collected nearly quantitatively on the same days between 08.00 and 20.00 hours over periods of 2 h. Trypsin and chymotrypsin activities in duodenal digesta did not differ between treatments at any time point (P > 0.05). In ileal digesta of pigs given diet HT the mean activity of trypsin was reduced (P < 0.05). The activity of chymotrypsin in ileal digesta did not differ between treatments. Trypsin activity:chymotrypsin activity was somewhat lower in ileal digesta of pigs receiving the HT diet (P < 0.10). The apparent ileal digestibility of crude protein (N x 6.25) was lower for the HT than for the LT diet (0.614 v. 0.728; P < 0.05). Condensed tannins are probably responsible for the lower activity of trypsin in ileal digesta of pigs fed on high-tannin faba bean hulls. Various explanations for the absence of effects of condensed tannins on enzyme activity in duodenal digesta are discussed.

Animals↗

Effect of tannins on starch digestibility and TMEn of triticale and semipurified starches from triticale and field beans.

1. The effects of field bean tannin-containing hulls (T-hulls), non-tannin hulls (NT-hulls) and tannin extract on starch digestibilities and TMEn of triticale and semipurified starches from triticale and field beans were evaluated with adult cockerels and 3-week-old-chicks. 2. For both cockerels and chicks, the digestibility and TMEn of semipurified starches from triticale were higher than those from beans. Starch digestibilities of triticales (range 99.1 to 97.7%), of semipurified starches from triticales (range 96.5 to 89.9%) and from field beans (range 75.4 to 71.6%) were similar for both cockerels and chicks. 3. The addition of NT-hulls improved, in general, the digestibility of semipurified starches for both chicks and cockerels, which suggests that fibre improves the digestion of semipurified starches. 4. The effects of tannins of T-hulls on semipurified starch from beans and from triticale were different. Tannins always decreased the digestibility and TMEn of semipurified starch from field beans but only decreased the digestibility and TMEn of semipurified starch from triticale when fed to chicks, but not when fed to cockerels. 5. The addition of tannin extract always decreased the digestibility of semipurified starches by a greater amount than T-hulls, which supports the idea that the effect of tannins depends on the quantity of tannin ingested.

Aging↗

Production of proline-rich proteins by the parotid glands of rats is enhanced by feeding diets containing tannins from faba beans (Vicia faba L.).

Feeding a casein-based diet with either 400 g/kg of tannin-containing faba bean hulls (Vicia faba L.) (1.41% condensed tannins) or 60 g/kg of a tannin-rich hull extract of faba beans (1.99% condensed tannins) to rats over a period of 7 d resulted in a 2.6 and 1.5 fold increase in weight of the parotid glands, respectively, (P < 0.05) and a corresponding 5.5 and 3.7 fold increase in the level of proline-rich proteins in the glands (P < 0.05). In a dose-response experiment, increasing the level of tannin-rich hull extract in the diet (0.0, 3.8, 7.5, 15.0, 30.0 and 60.0 g/kg) resulted in a linear increase in both the relative size of parotid glands (R2 = 0.90; P < 0.05) and the quantity of proline-rich proteins in the glands (R2 = 0.89; P < 0.05). The apparent digestibility of total (R2 = 0.97) and individual amino acids (R2 varied from 0.27 to 0.99) decreased linearly (P < 0.05). The quantity of proline-rich proteins in the cecum of rats was estimated from the decrease in digestibility of proline, glycine and glutamic acid. The estimated secretions of proline-rich proteins, when calculated on the basis of the three respective amino acids, were 3.5, 3.5 and 3.9 mg of proline-rich proteins per 10 mg of additional hull extract (21.8% condensed tannins). The results indicate that tannins from faba beans stimulate the parotid glands to increase the secretion of proline-rich proteins in rats. The proline-rich proteins then interact with dietary condensed tannins to reduce their antinutritional effects.

Amino Acids↗

Effect of condensed tannins extracted from four forages on the viability of the larvae of deer lungworms and gastrointestinal nematodes.

The inhibitory activity of condensed tannins extracted from four forage legume plants were evaluated by using a larval migration inhibition assay. The first (L1) and third (L3) stages of deer lungworm (Dictyocaulus viviparus), and the third stage (L3) of deer gastrointestinal nematodes were incubated with tannins extracted from Lotus pedunculatus, Lotus corniculatus, sulla (Hedysarum coronarium) and sainfoin (Onobrychus viciifolia). The tannins extracted from all the forages had inhibitory activity as measured by their ability to paralyse the larvae and inhibit them from passing through sieves. At the highest concentration used (1200 microg/ml) the tannins extracted from sainfoin had the highest activity against ensheathed L1 lungworm larvae (58 per cent), followed by L. pedunculatus (45 per cent), sulla (42 per cent) and L. comiculatus (35 per cent) when the larvae were incubated at 37 degrees C. The same trend, but with lower activities, was observed when the larvae were incubated at 22 degrees C. Anthelmintic activity against L3 lungworm larvae was evaluated by measuring the death rate of ensheathed L3 larvae after incubation with condensed tannins for two, 24 and 48 hours at room temperature (22 degrees C). The death rate was significantly higher (P<0.001) after 48 hours incubation than after two hours or 24 hours, and significantly higher (P<0.001) after 24 hours than after two hours incubation. Condensed tannins from sainfoin had the highest inhibitory activity followed by L. pedunculatus, sulla and L. comiculatus. The tannins from sainfoin also had the highest activity against L3 larvae of gastrointestinal nematodes, followed by L. pedunculatus, sulla and L. comiculatus. Exsheathed larvae of gastrointestinal nematodes were significantly more susceptible to the action of the tannins than ensheathed larvae.

Animals↗

Dietary grape seed tannins: effects of nutritional balance and on some enzymic activities along the crypt-villus axis of rat small intestine.

The aim of the study was to determine the nutritional and intestinal effects of grape seed tannins. For this purpose, tannins were incorporated in diets of rats at levels of 0.2 or 2.0% for 31 days in comparison to a control diet. The animals were pair-fed. Nutritional balances were not affected by feeding 0.2% tannins. At the highest dose (2%) grape seed tannins reduced growth as well as dry matter (DM) and nitrogen (N) digestibility. In rats fed protein-free diets, 2% tannins significantly increased endogenous fecal N. Starch and fat were well digested in all groups of rats. No changes in organ weights were observed. Duodenal alkaline phosphatase activity (AP) was never affected by tannins. On the other hand, in the jejunum, along the vilus-crypt unit, a reduction of AP and sucrase appeared at the tip villus which was balanced by an enhancement of 3H-thymidine incorporation in the middle of the crypt zone, giving evidence of endogenous N loss. This study did not reveal a major toxic effect of tannins except a reduced DM and N digestibility; nevertheless tannins directly interfere with mucosal proteins, thereby stimulating the cell renewal.

Alkaline Phosphatase↗

Plant tannins inhibit the induction of aberrant crypt foci and colonic tumors by 1,2-dimethylhydrazine in mice.

We have shown that naturally occurring tannins possess antitumor promotion activity in mouse skin. In the present investigation, we studied the ability of a hydrolyzable tannin, gallotannin (GT), and a condensed tannin extracted from red alder (RA) bark to inhibit 1,2-dimethylhydrazine (DMH)-induced colonic aberrant crypt foci (ACF) and tumors in Balb/c mice. In addition, we determined the ability of GT to inhibit the proliferation and to induce apoptosis in a human colon cancer cell line (T-84). Mice were given tannins by intraperitoneal injections, by gavage, or in drinking water before treatment with DMH for 24 weeks. Alternatively, mice were given tannins by intraperitoneal injection or gavage for only 2 weeks before DMH administration, then tannin administration was discontinued and mice were treated with DMH for 24 weeks. The multiplicity, size, and distribution of ACF and tumors were significantly inhibited by GT and RA in the above treatment regimens. The most effective treatments included GT by gavage, RA bark extract by intraperitoneal injection, and either tannin dissolved in drinking water. Extent of inhibition of ACF and tumors was gender independent. In cell culture experiments, GT treatment for three days inhibited the growth of T-84 cells, with a concentration resulting in half-maximal inhibition estimated to be 20 micrograms/ml. The treatment was not cytotoxic to cells at 1-40 micrograms/ml. Interestingly, at 10 micrograms/ml, GT induced apoptosis in T-84 cells as determined by the Hoechst DNA staining technique. Collectively, these findings support a potential role for tannins as chemopreventive agents against colon cancer.

1,2-Dimethylhydrazine↗

Investigations of leg abnormalities in chicks consuming high tannin sorghum grain diets.

Studies were conducted into the etiology of leg abnormalities noted in chicks fed high tannin sorghum grain diets. These anomalies were characterized by a bowing of the legs with a swelling of the hock joints. The incidence of these leg problems was found to be markedly higher when the amino acids in the diet provided by soybean meal were replaced by crystalline amino acids. Supplemental vitamins and minerals had no alleviating effect on the leg problem. Bone mineralization was apparently not influenced by tannins as demonstrated by similar bone ash values for chicks fed high or low tannin sorghums. A possible alteration caused by tannin in the organic matrix of bone is discussed. High tannin sorghum depressed chick growth and feed conversion when compared with low tannin sorghum in both sorghum-soybean meal and sorghum-amino acid rations. This growth depression was overcome by supplementing the high tannin sorghum-soybean meal diet with .15% DL-methionine, but no growth response was observed from a similar supplementation of a high tannin sorghum-amino acid diet.

Amino Acids↗

[Separation of tannins in Rhubarb and its analysis by high performance liquid chromatography-mass spectrometry].

In order to investigate the pharmaceutical actions of rhubarb, a method for extracting, separating and analyzing the tannin components in rhubarb was studied. At first, a procedure for the group separation of tannins from the water-ethanol extract of rhubarb was established based on the formation of tannins-caffein precipitation. Then, a high performance liquid chromatographic (HPLC) method for the analysis of tannins in rhubarb was developed. This HPLC method is based on a reversed-phase C18 column and polar mobile-phase such as water and methanol with gradient elution, and the tannins can be well separated. Finally, the identification of the tannin components in rhubarb was carried out by high performance liquid chromatography-mass spectrometry (HPLC-MS). The structures of the main tannin components (gallic acid, catechin, the dimer, trimer, tetramer and pentamer of catechin) in rhubarb are suggested. The fragmentation laws of the tannin components are summarized. In comparing with previous methods, it is simple and without the interference of co-existed compounds.

Chromatography, High Pressure Liquid↗

Adsorptive removal of tannin from aqueous solutions by cationic surfactant-modified bentonite clay.

The removal of tannin from aqueous media by cationic surfactant-modified bentonite clay was studied in a batch system. The surfactant used was hexadecyltrimethylammonium chloride. Adsorbent characterizations were investigated using X-ray diffraction, infrared spectroscopy, surface area analysis, and potentiometric titration. The effects of pH, contact time, initial solute concentration, adsorbent dose, ionic strength, and temperature on the adsorption of tannin onto modified clay were investigated. The adsorbent exhibited higher tannin removal efficiency (>99.0%) from an initial concentration of 10.0 micromol/L at pH 3.0. Adsorption capacity decreased from 90.1 to 51.8% with an increase in temperature from 10 to 40 degrees C at an initial concentration of 25.0 micromol/L. The adsorption process was found to follow pseudo-first-order kinetics. Film diffusion was found to be the rate-limiting step. Tannin adsorption was found to decrease with increase in ionic strength. The tannin equilibrium adsorption data were fitted to Langmuir and Freundlich isotherm models, the former being found to provide the best fit of the experimental data. The maximum monolayer adsorption capacity for tannin was 69.80 micromol/g at 30 degrees C. Comparison of adsorption capacity of the modified clay with reported adsorbents in the literature was also presented. Adsorbed tannin on modified clay can be recovered by treatment with 0.1 M NaOH solution. Regeneration experiments were tried for four cycles and results indicate a capacity loss of <10.0%. From the results it can be concluded that the surfactant-modified clay could be a good adsorbent for treating tannin-contaminated waters.

Journal Article↗

Effects of Condensed Tannins on Endoglucanase Activity and Filter Paper Digestion by Fibrobacter succinogenes S85.

The effect of condensed tannins from birdsfoot trefoil (Lotus corniculatus L.) on the cellulolytic rumen bacterium Fibrobacter succinogenes S85 was examined. Condensed tannins inhibited endoglucanase activity in the extracellular culture fluid, at concentrations as low as 25 mug ml. In contrast, cell-associated endoglucanase activity increased in concentrations of condensed tannins between 100 and 300 mug ml. Inhibition of endoglucanase activity in both the extracellular and the cell-associated fractions was virtually complete at 400 mug of condensed tannins ml. Despite the sharp decline in extracellular endoglucanase activity with increasing concentrations of condensed tannins, filter paper digestion declined only moderately between 0 and 200 mug of condensed tannins ml. However, at 300 mug ml, filter paper digestion was dramatically reduced and at 400 mug ml, almost no filter paper was digested. F. succinogenes S85 was seen to form digestive grooves on the surface of cellulose, and at 200 mug ml, digestive pits were formed which penetrated into the interior of cellulose fibers. Cells grown with condensed tannins (100 to 300 mug ml) possessed large amounts of surface material, and although this material may have been capsular carbohydrate, its osmiophilic nature suggested that it had arisen from the formation of tannin-protein complexes on the cell surface. The presence of electron-dense extracellular material suggested that similar complexes were formed with extracellular protein.

Journal Article↗

Antibacterial activity of hydrolyzable tannins derived from medicinal plants against Helicobacter pylori.

Helicobacter pylori is a major etiological agent in gastroduodenal disorders. In this study, we isolated 36 polyphenols and 4 terpenoids from medicinal plants, and investigated their antibacterial activity against H. pylori in vitro. All hydrolyzable tannins tested demonstrated promising antibacterial activity against H. pylori. Monomeric hydrolyzable tannins revealed especially strong activity. Other compounds demonstrated minimal antibacterial activity with a few exceptions. A monomeric hydrolyzable tannin, Tellimagrandin I demonstrated time- and dose-dependent bactericidal activity against H. pylori in vitro. On the other hand, hydrolyzable tannins did not affect the viability of MKN-28 cells derived from human gastric epithelium. Hydrolyzable tannins, therefore, have potential as new and safe therapeutic regimens against H. pylori infection. Furthermore, we investigated effects of hydrolyzable tannins on lipid bilayer membranes. All the hydrolyzable tannins tested demonstrated dose-dependent membrane-damaging activity. However, it remains to be elucidated whether their membrane-damaging activity directly contributes to their antibacterial action.

Acids↗

Effect of nitrogen fixation, nitrogen fertilization and viral infection on yield, tannin and protein contents and in vitro protein digestibility of faba bean.

A field investigation of two faba bean cultivars (cv.), Agabat and Silaim, showed that bean yellow mosaic virus (BYMV) infection reduced (p < or = 0.001) yield (Kg/ha), protein content and in vitro protein digestibility (IVPD) but increased (p < or = 0.05) tannin content (mg/100 ml). Nitrogen fertilization with viral infection significantly reduced yield and IVPD for cv. Silaim and increased (p < or = 0.05) protein and tannin contents. Nitrogen fertilization alone was found to increase (p < or = 0.05) yield, protein and tannin contents but slightly reduced IVPD. Rhizobium inoculation with viral infection significantly decreased yield per unit area, protein content and IVPD, but increased (p < or = 0.05) tannin content. Rhizobium inoculation alone significantly increased (p < or = 0.001) yield and tannin content and slightly increased protein content but decreased IVPD. The results indicated that nitrogen fertilization or nitrogen fixation increased yield, protein and tannin contents and decreased IVPD. Viral infection had an adverse effect on yield, protein content and IVPD but had no effect on tannin content.

Digestion↗

In vitro availability of iron and zinc in white and coloured ragi (Eleusine coracana): role of tannin and phytate.

White and brown ragi (Eleusine coracana) varieties were analysed for tannin, phytate phosphorus, total phosphorus, iron, ionisable iron, zinc and soluble zinc content. White ragi had no detectable tannin while in brown varieties it ranged from 351 to 2392 mg per 100 g. Germination brought about a progressive decrease in tannin and phytate phosphorus and an increase in ionisable iron and soluble zinc content of grain ragi. Both in raw and germinated grain, ionizable iron was significantly higher in white than in brown varieties. While ionisable iron was inversely correlated with the level of tannin and phytate phosphorus, soluble zinc was negatively correlated with phytate phosphorus. After extraction of tannin, ionisable iron of brown ragi rose by 85%. On the other hand, in white varieties, addition of tannin extracted from brown ragi, resulted in a 52-65% decrease in ionisable iron content. These studies indicated that poor iron availability in ragi as judged by its low ionisable iron content was due to the presence of tannin in the grain.

Biological Availability↗

Protein-binding capacity of microquantities of tannins.

The physiological effect of tannins is studied in terms of their protein-binding or precipitation capacity. A number of assays based on binding of hemoglobin or bovine serum albumin (BSA) and subsequent determination of unbound protein in supernatant or tannin in a protein-tannin complex are available but with various limitations. These methods are unable to estimate protein-binding capacity, if the quantity of tannin available is low. In the method reported here, tannins or other phenolics were applied on chromatography paper and reacted with BSA and unbound BSA was washed off. The protein in the tannin-protein complex was measured spectrophotometrically after staining with Ponceau S. It required microquantities of sample. Using this method the protein-binding capacity of total leaf extract and hydrolyzable and condensed tannins of Quercus incana, Q. semecarpofolia, and Q. dilatata was determined. The protein binding capacities of ellagic acid and quercetin (microgram BSA/mg) were 297.3 and 78.0, respectively.

Hydrolysis↗

Response of DNA, proteins and membrane bilayer in the digestive gland cells of freshwater mussel Unio tumidus to tannins exposure.

Exposure of digestive gland cells (Unio tumidus) in in vitro study to tannins (tannic, ellagic and gallic acid) caused changes in three types of cellular macromolecules, i.e. lipids, proteins and DNA. Two fluorescence probes: TMA-DPH and 12-AS were used to assess the membrane fluidity after incubating cells with different concentrations of tannins (1-60 microM). It was noticed that tannins increased the fluidity in the internal region of the lipid bilayer, but no changes at the surface of the plasma membrane were observed. The number of protein-bound carbonyl groups was quantitated spectrophotometrically using 2,4-dinitrophenylhydrazine (DNPH) assay. Oxidative modification of proteins increased in the cells exposed to tannins in a concentration-dependent manner. Results also showed an increase in carbonyl groups formation after exposing the cells to complex: tannin-Cu(2+) ions. We also investigated the effect of tannins on the level of nuclear DNA damage using the comet assay. We showed that these compounds at 15, 30 and 60 microM induced DNA damage. At these concentrations cell viability measured by trypan blue exclusion assay was not reduced by more than 30%. We also added Cu(2+) ions at 50 microM to assess the influence of the complex tannin-Cu(2+) on DNA lesions. The same method was used to study the possibility to repair this damage in DNA. Experimental data showed that tested polyphenolic acids do not contribute to the induction of strand breaks in nucleic acid at the smallest dose--1 microM and in this range they have a protective effect against the activity of 50 microM Cu(2+) ions.

Animals↗

Structural determinants of the platelet agonist activity of cotton bract condensed tannin.

Previous studies on the platelet agonist activity of cotton bract condensed tannin have used tannins with apparent molecular weights greater than 10,000 Da as measured by dialysis and/or ultrafiltration. However, because tannins in cotton dust are heterogeneous in terms of molecular weight, it was important to determine if platelet agonist activity was related to polymer length. To accomplish this, aqueous extracts of cotton dust were fractionated by a series of graded Amicon ultrafiltrations and the resulting fractions were examined for tannin content and platelet agonist activity. All tannins with apparent molecular weight greater than 1000 Da were equally potent platelet agonists while those less than 1000 Da had no measurable platelet agonist activity. This polymer length dependence for activity was supported by the observation that two procyanidin dimers had only minimal platelet agonist activity. In addition, methylation of the tannin hydroxyl groups resulted in the loss of platelet agonist activity. Thus, the platelet agonist activity of tannin requires a minimum polymer length and the presence of free hydroxyl groups.

Blood Platelets↗

Tannins in tropical browses: effects on in vitro microbial fermentation and microbial protein synthesis in media containing different amounts of nitrogen.

Four species of browses (Acacia angustissima, Acacia salicina, Calliandra calothyrsus, andDichrostachys cinerea) were used to study the effect of tannins on microbial fermentation and microbial protein synthesis in incubation media containing high nitrogen (HN) and low nitrogen (LN) in the presence and absence of polyethylene glycol (PEG, MW 6000). The additional nitrogen in HN medium was supplied through ammonium bicarbonate. The use of HN medium significantly (P < 0.05) increased the in vitro gas and short-chain fatty acid (SCFA) production and microbial protein synthesis compared to the LN medium. Incubation of tannin-containing browses alone produced significantly (P < 0.05) lower gas and SCFA compared to in the presence of PEG in both HN and LN media. Inclusion of PEG in tannin-containing browses significantly (P < 0.05) reduced the molar proportion of propionate compared to in its absence. Higher N in the media resulted in 10.4 and 9.9% increases in in vitro gas and SCFA production, respectively, whereas inclusion of PEG to tannin-containing feed to remove the effect of tannins increased the in vitro gas and SCFA production by 186 and 195%, respectively, indicating that the low fermentation of tannin-containing browses could be due to the depressive effects of tannins on microbial activity and only partially accounted for by unavailability of N for rumen microbes. Incubation of browses with straw significantly (P < 0.05) decreased ammonia nitrogen concentration but increased the in vitro gas and SCFA production and microbial protein synthesis compared to straw alone.

Bacterial Proteins↗