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Complexing of toxic hydrolysable tannins of yellow-wood (Terminalia oblongata) and harendong (Clidemia hirta) with reactive substances: an approach to preventing toxicity.

Ruminants consuming either tannic acid or hydrolysable tannins from the Australian yellow-wood tree (Terminalia oblongata) and the Indonesian shrub Clidemia hirta are intoxicated by simple phenolics liberated in the gut. The affinity of these tannins and of the simple phenolic gallic acid for the two proteins casein and pepsin, polyvinylpyrolidone (PVP), activated charcoal and Ca(OH)2 was examined in vitro. The studies were undertaken to predict the effect of these phenolics on digestion and to identify substances that would act as antidotes by precipitating phenolics. Tannins but not gallic acid were precipitated as stable complexes with both pepsin and casein at pH 3-5. Optimal complexing of tannin with protein occurred at a weight ratio of 1:1. Ionic strength and temperature did not affect the amount of tannin precipitated from solution with protein. The precipitation of tannins with PVP and Ca(OH)2 was unaffected by pH within the range 2-8 while maximum binding with activated charcoal occurred between pH 3 and 7. In contrast to protein, the other substances complexed with gallic acid; only gallic acid-PVP complexes were affected by pH. Calcium hydroxide bound more tannin and gallic acid on a weight basis than PVP and charcoal. Both Ca(OH)2 and activated charcoal should complex with phenolics in the forestomach, abomasum and intestines. The reaction of hydrolysable tannins and proteins at the pH found in the abomasum suggests that hydrolysable tannins would interfere with enzyme function and protein digestion post-ruminally rather than in the forestomach.

Animals↗

Effects of tannins from different sources on airway epithelial and platelet function.

The uniqueness of tannin isolated from aqueous extracts of cotton bracts (CBE) on the electrophysiological and ion transport properties of the canine tracheal epithelium and on 5-hydroxytryptamine (5-HT) release from platelets was investigated. The effect of CBE from Deltapine 41 cotton picked before (green, containing high-concentration, high-molecular-weight tannin) and after (brown, containing low-concentration, low-molecular-weight tannin) senescence and Darjeeling tea extracts and tannin isolated from Darjeeling tea were compared to the effects of Acala SJ-2 CBE and Acala SJ-2 tannin. Green Deltapine 41 CBE was similar to Acala SJ-2 CBE in decreasing short-circuit current (Isc) and net chloride secretion and in producing a dose-response release of 5-HT from platelets. Both Green and Brown Deltapine 41 CBE altered the paracellular pathway, but were less potent than Acala SJ-2 CBE. Brown Deltapine 41 CBE stimulated chloride transport in the airway epithelium and did not produce 5-HT release from human platelets. Darjeeling tea and tea tannin increased Isc and net chloride secretion and caused 5-HT release from platelets only at high concentrations. These studies demonstrate that all tannins are not alike; they suggest that molecular weight and concentration may be important for tannin activity and that the tannin in cotton bracts extract may be unique to cotton in its effect on the airway epithelium and on platelets.

Airway Resistance↗

Interaction of gut microflora with tannins in feeds.

Tannins (hydrolyzable and condensed) are water-soluble polyphenolic compounds that exert antinutritional effects on ruminants by forming complexes with dietary proteins. They limit nitrogen supply to animals, besides inhibiting the growth and activity of ruminal microflora. However, some gastrointestinal microbes are able to break tannin-protein complexes while preferentially degrading hydrolyzable tannins (HTs). Streptococcus gallolyticus, Lonepinella koalarum and Selenomonas ruminantium are the dominant bacterial species that have the ability to degrade HTs. These tanninolytic microorganisms possess tannin-degrading ability and have developed certain mechanisms to tolerate tannins in feeds. Hence, selection of efficient tanninolytic microbes and transinoculation among animals for long-term benefits become areas of intensive interest. Here, we review the effects of tannins on ruminants, the existence and significance of tannin-degrading microorganisms in diverse groups of animals and the mechanisms that tannin-degrading microorganisms have developed to counter the toxic effects of tannin.

Animal Feed↗

Adsorption mechanism of hexavalent chromium by redox within condensed-tannin gel.

We have proposed a new recovery system of hexavalent chromium Cr(VI) that is of great toxicity utilizing condensed-tannin gels derived from a natural polymer with many polyhydroxyphenyl groups. The adsorption mechanism of Cr(VI) to the tannin molecules was clarified. The adsorption mechanism consists of four reaction steps; the esterification of chromate with tannin molecules, the reduction of Cr(VI) to trivalent chromium Cr(III), the formation of carboxyl group by the oxidation of tannin molecules and the ion exchange of the reduced Cr(III) with the carboxyl and hydroxyl groups. It was found in this recovery system that a large amount of proton was consumed accompanied with the reduction of Cr(VI) so that the acidic solution containing Cr(VI) was transferred automatically to neutral one by choosing an appropriate initial pH. The carboxyl group which was created by the oxidation of tannin molecules parallel to the reduction of Cr(VI) to Cr(III) contributed to an increase in the ion-exchange sites of the reduced Cr(III). The maximum adsorption capacity of Cr(VI) reached 287 mg Cr/g dry tannin gel under the conditions of 0.77 water content of tannin gel and the initial pH = 2. This adsorption capacity was five to ten times higher than that obtained by the ion exchange between ordinary Cr(III) and tannin molecules for the tannin gels prepared under similar conditions. The system proposed here will provide an important information on a zero-emission-oriented process because it has such advantages as higher adsorption capacity of chromium and lower volume of secondary wastes compared with conventional process.

Adsorption↗

A novel method to evaluate the relative tannin-binding capacities of salivary proteins.

Many herbivore species have a diet containing high proportions of polyphenolics, principally lignins and tannins; the latter reduce the conversion of ingested nutrients into biomass and exert systemic toxicity at high levels of intake. It has been proposed that tannin-binding proteins in the saliva might be responsible for minimizing these tannin-related effects by forming soluble protein-tannin complexes. We have developed a method that permits evaluation of the relative tannin-binding properties of salivary proteins at a pH of 8.0-8.5. It is aimed at facilitating both the identification of tannin-binding proteins and the investigation of their relative tannin binding. The principle of the assay is the inhibition of trypsin by tannins and the subsequent reversal of that inhibition when other tannin-binding proteins are added (indirect assay). The method is rapid; large sample numbers may be processed by virtue of its microtiterplate format.

Gelatin↗

Linking chemical reactivity and protein precipitation to structural characteristics of foliar tannins.

Tannins influence ecosystem function by affecting decomposition rates, nutrient cycling, and herbivory. To determine the role of tannins in ecological processes, it is important to quantify their abundance and understand how structural properties affect reactivity. In this study, purified tannins from the foliage of nine species growing in the pygmy forest of the northern California coast were examined for chemical reactivity, protein precipitation capacity (PPC), and structural characteristics (13C NMR). Reactivity of purified tannins varied among species 1.5-fold for the Folin total phenol assay, and 7-fold and 3-fold, respectively, for the acid butanol and vanillin condensed tannin assays. There was about a 5-fold difference in PPC. Variation in chemical reactivity and PPC can be largely explained by differences in structural characteristics of the tannins determined by 13C NMR. In particular, the condensed versus hydrolyzable tannin content, as well as the hydroxylation pattern of the B-ring and stereochemistry at the C-2-C-3 position appear to influence reactivity. Due to the large differences in chemical reactivity among species, it is necessary to use a well-characterized purified tannin from the species of interest to convert assay values to concentrations. Our results suggest that structural characteristics of tannins play an important role in regulating their reactivity in ecological processes.

Animals↗

The role of condensed tannins in the nutritional value of Lotus pedunculatus for sheep. Rates of body and wool growth.

Lotus pedunculatus (cv. Grasslands Maku) grown on acid low-fertility soil and containing high concentrations of condensed tannin (76-90 g/kg dry matter (DM] was grazed by growing sheep for 31-42 d periods in three experiments. In Expt 2 an additional group of lambs grazed areas oversown with white clover (Trifolium repens) and red clover (Trifolium pratense). Lambs were transferred from grazing ryegrass (Lolium perenne)-white clover straight on to lotus in all experiments (unconditioned sheep). In Expt 3 a second group was included which had grazed high-tannin lotus for a pre-experimental period of 8 weeks (conditioned sheep). Effects of condensed tannin on body and wool growth were assessed by studying responses to daily oral administration of polyethylene glycol (PEG; molecular weight 3350, 75-100 g/d). PEG forms a complex with condensed tannin, which is assumed to be inert in its passage through the digestive system, and so effectively reduces the nutritional effects attributable to high condensed-tannin concentrations. Live-weight gain (LWG) in the absence of PEG was low (27-125 g/d) for sheep grazing high-tannin lotus, and PEG administration increased LWG by 41-61 g/d and increased wool growth. In Expt 3, responses to PEG supplementation tended to be less with conditioned than with unconditioned sheep, indicating that conditioned sheep had partially adapted to the high-tannin diet. PEG supplementation had no effect on either LWG or wool growth of sheep grazing areas oversown with mixed clovers, confirming its effects as specific to forages containing condensed tannins. These experiments therefore conclusively show that high concentrations of condensed tannin induced by growing Lotus pedunculatus under low soil fertility conditions prevent maximum expression of LWG and wool growth in grazing sheep. These results contrast with high LWG (153-315 g/d) observed in growing sheep grazing the same lotus cultivar grown in high fertility soil and containing 20 g condensed tannin/kg DM, a level considered to be nutritionally beneficial.

Administration, Oral↗

Effects of condensed tannins prepared from leaves of fodder plants on digestive enzymes in vitro and in the intestine of rats.

1. Of the nineteen plants screened, six were found to contain large quantities of condensed tannins. Black locust (Robinia pseudo-Acacia), bush clover (Lespedeza bicolor), wistaria (Wistaria floribunda) and Japanese knotgrass (Reynoutria japonica) were used for the present experiment. Tannins of the investigated plants were fractionated into three or four molecular forms, according to the degree of polymerization, by chromatography on a column of Sephadex LH-20. 2. The protein-precipitating capacity of the fractionated tannins increased with the increase in degree of polymerization. The inhibitory effect of tannins on trypsin (EC 3.4.21.4), alpha-amylase (EC 3.2.1.1) and lipase (EC 3.1.1.3) activities in vitro also increased with the increase in degree of polymerization. The digestion of tannin-bovine serum albumin complex by trypsin was related to the degree of polymerization of tannins complexed. 3. Inclusion of black locust tannins in the diet (10 g/kg) depressed the activities of trypsin and alpha-amylase in the upper, middle and lower parts of the intestine of the rats, but the lipase activity was increased in the middle part and remained unaffected in the upper and lower parts. It is presumed that the tannins have little affinity for lipase. 4. Digestion trials, when the microflora level of the alimentary canal of rats was reduced by dietary antibiotic treatment, revealed that the inclusion of tannins in the diet (20 g/kg) depressed the digestibilities of proximate constituents, except crude fat, and increased faecal bile acid excretion.

Amylases↗

Effect of tannin-rich leaves of oak (Quercus incana) on various microbial enzyme activities of the bovine rumen.

1. The objective of the present experiment was to study the effects of oak (Quercus incana) leaves rich in tannins on various enzyme activities of the bovine rumen. 2. The procedure employed was incubation of tannin-rich, very-low-tannin or virtually tannin-free leaves in nylon-gauze bags in the rumen, and determination of enzyme activities in microbes tightly bound to the solid matrix and in microbes loosely plus tightly attached to the solid matrix. 3. The activities of urease (EC 3.5.1.5), carboxymethylcellulose, glutamate dehydrogenase (EC 1.4.1.2) and alanine aminotransferase (glutamic-pyruvic transaminase) (EC 2.6.1.2) were significantly lower in the tannin-rich group, whereas the activities of glutamate ammonia ligase (glutamine synthetase) (EC 6.3.1.2; both gamma-glutamyltransferase (EC 2.3.2.2) and the forward reaction) were higher in the tannin-rich group. These changes were more marked in micro-organisms tightly bound to the solid matrix than in the more complex microbial compartment. 4. The protein, DNA and RNA contents, and protein: RNA ratio, were significantly lower in the tannin-rich group, whereas no difference was observed for protein: DNA between the groups. 5. Effects of tannin-containing extracts of oak leaves on various rumen enzymes in vitro showed a trend similar to that observed in nylon-gauze bags, suggesting that the changes observed in various compartments were due to the tannins of oak leaves.

Alanine Transaminase↗

Lotus corniculatus condensed tannins decrease in vivo populations of proteolytic bacteria and affect nitrogen metabolism in the rumen of sheep.

Condensed tannins in forage legumes improve the nutrition of sheep by reducing ruminal degradation of plant protein and increasing crude protein flow to the intestine. However, the effects of condensed tannins in forage legumes on rumen bacterial populations in vivo are poorly understood. The aim of this study was to investigate the specific effects of condensed tannins from Lotus corniculatus on four proteolytic rumen bacteria in sheep during and after transition from a ryegrass (Lolium perenne)-white clover (Trifolium repens) diet (i.e., low condensed tannins) to a Lotus corniculatus diet (i.e., higher condensed tannins). The bacterial populations were quantified using a competitive polymerase chain reaction. Lotus corniculatus was fed with or without ruminal infusions of polyethylene glycol (PEG), which binds to and inactivates condensed tannins, enabling the effect of condensed tannins on bacterial populations to be examined. When sheep fed on ryegrass-white clover, populations of Clostridium proteoclasticum B316T, Butyrivibrio fibrisolvens C211a, Eubacterium sp. C12b, and Streptococcus bovis B315 were 1.5 x 10(8), 1.1 x 10(6), 4.6 x 10(8), and 7.1 x 10(6) mL(-1), respectively. When the diet was changed to Lotus corniculatus, the average populations (after 8-120 h) of C. proteoclasticum, B. fibrisolvens, Eubacterium sp., and S. bovis decreased (P < 0.001) to 2.4 x 10(7), 1.1 x 10(5), 1.1 x 10(8), and 2.5 x 10(5) mL(-1), respectively. When PEG was infused into the rumen of sheep fed Lotus corniculatus, the populations of C. proteoclasticum, B. fibrisolvens, Eubacterium sp., and S. bovis were higher (P < 0.01-0.001) than in sheep fed Lotus corniculatus without the PEG infusion, with average populations (after 8-120 h) of 4.9 x 10(7), 3.8 x 10(5), 1.9 x 10(8), and 1.0 x 10(6), respectively. Sheep fed the Lotus corniculatus diet had lower rumen proteinase activity, ammonia, and soluble nitrogen (P < 0.05-0.001) than sheep that were fed Lotus corniculatus plus PEG. The Lotus corniculatus diet reduced rumen nitrogen digestibility (P < 0.05) and ammonia pool size and increased the flow of undegraded feed nitrogen to the abomasum. The nitrogen intake, rumen non-ammonia nitrogen pool size, rumen microbial non-ammonia nitrogen pool size, and abomasal microbial non-ammonia nitrogen fluxes were similar both in sheep fed only Lotus corniculatus and in sheep fed Lotus corniculatus plus PEG, but nonmicrobial non-ammonia nitrogen flux to the abomasum was higher (P < 0.01) for the sheep fed only Lotus corniculatus. Although condensed tannins in Lotus corniculatus reduced the populations of some proteolytic bacteria, total ruminal microbial protein and microbial protein outflow to the abomasum were unchanged, suggesting a species-specific effect of condensed tannins on bacteria in the rumen.

Animals↗

Calcium influx is required for tannin-mediated arachidonic acid release from alveolar macrophages.

The role of Ca2+ was investigated in the response of alveolar macrophages to cotton tannin, an agent implicated in the lung disease byssinosis in textile mill workers. A physiological concentration of extracellular Ca2+ was found to be required for tannin-mediated release of radiolabeled arachidonic acid (AA). Flow cytometry using indo 1 indicated that tannin caused a rapid and dose-dependent Ca2+ increase in macrophages that also required extracellular Ca2+. Ethylene glycol-bis(beta-aminoethyl ether)-N,N,N',N'-tetraacetic acid virtually abolished the Ca2+ influx mediated by tannin but had little effect on intracellular Ca2+ release induced by thapsigargin, N-formylmethionyl-leucylphenylalanine, or thimerosal. A mechanism for extracellular Ca2+ influx was demonstrated by rapid Mn2+ quenching of indo 1 by tannin. Verapamil inhibited tannin-mediated Ca2+ influx and AA release, but the effective concentration was 100 microM. 1,2-Bis(2-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid chelated all Ca2+ in the cells and effectively abolished the tannin response. Exposure to tannin was not associated with cytotoxicity, as judged by 51Cr release. The data suggest that tannin induces Ca2+ influx in alveolar macrophages, which represents an important prerequisite for a cell-signaling pathway resulting in the accumulation of free AA.

Animals↗

Induction of interleukin-1-beta release from human monocytes by cotton bract tannin.

The human T lymphocyte proliferative response to cotton bract tannin was shown to be dependent upon the presence of monocytes. Since monocytes support the T cell mitogenic response by interleukin-1 (IL-1) production, it was anticipated that tannin has IL-1-inducing ability. To examine this possibility, human monocytes were cultured alone or with peripheral blood T lymphocytes, and stimulated with tannin. Control cultures included unstimulated cells, and cells challenged with other IL-1 inducers: concanavalin A (Con A) and lipopolysaccharide from Escherichia coli or Enterobacter agglomerans. IL-1 beta was measured in culture supernatants 24 h after initiation of the culture by the use of an ELISA or an RIA. The results showed that tannin stimulated monocytes to secrete IL-1 beta in a manner similar to Con A, i.e. substantially more cytokine was measured in the supernatants of monocyte-T-lymphocyte co-cultures than in the cultures of monocyte alone. Endotoxin from E. coli was less effective than the endotoxin from E. agglomerans in IL-1 induction. Contaminating endotoxin present in the tannin preparation accounted for the majority of IL-1 beta released from monocytes alone stimulated with tannin, but only 20% of the IL-1 beta released from tannin-stimulated monocyte-T-lymphocyte co-cultures. These results show that tannin itself has IL-1-inducing ability. The dose-response studies show that the extent of IL-1 beta release is dependent on tannin dose and that increased levels of monocyte-produced IL-1 beta precede the increase in T lymphocyte proliferation.(ABSTRACT TRUNCATED AT 250 WORDS)

Cells, Cultured↗

Effect of grape seed tannins on the activity of some rat intestinal enzyme activities.

The present study was designed to investigate the effects of grape seed tannins on rat intestinal alkaline phosphatase (AP), sucrase and dipeptidyl peptidase IV (DPP IV) activities. An experiment was performed in vivo by dietary supplementation with 2% tannins; this diet was tested on an experimental group of rats; a control group received a diet without tannins. After 31 days, tannins intake significantly decreased middle-jejunal AP from 123 to 45 mU/mg protein and sucrase activities from 310 to 195 mU/mg protein, while no significant difference appeared at the duodenal stage (p < 0.05). Ileal DPP IV activity was also significantly reduced (p < 0.05) from 190 to 110 mU/mg protein after tannin intake. Using in vitro experiments on purified brush border membranes, AP activity was found to be inhibited by grape tannins; this inhibition was prevented by the detergent Triton X-100. The addition of pancreatic-biliary (PB) juice to the incubation medium prevented or reversed the tannin-inhibited enzyme activity. The present data indicate that in the duodenal lumen, alkalinity and detergency from the PB secretion neutralized the ability of tannins to inactivate brush border hydrolase activities and suggest that enzyme inhibition took place once bile salts were reabsorbed while moving down the gut. This was confirmed by in vitro experiments where sucrase and DPP IV activities inhibited by grape seed tannins were largely recovered after the addition of PB juice to the incubation medium.

Alkaline Phosphatase↗

Nutritional toxicology of tannins and related polyphenols in forage legumes.

Proanthocyanidins (PA) (condensed tannins) and hydrolyzable tannins (HT) are the two major classes of tannins. Proanthocyanidins are flavonoid polymers. Hydrolyzable tannins are polymers of gallic or ellagic acid esterified to a core molecule, commonly glucose or a polyphenol such as catechin. Proanthocyanidins are the most common type of tannin found in forage legumes. Problems in the analysis of tannins are that sample processing and drying decrease extraction and reactivity, suitable standards are unavailable, and quantitative analytical methods are poorly correlated with enzyme inhibition, protein precipitation, and nutritional effects. Hydrolyzable tannins are potentially toxic to ruminants. Pyrogallol, a hepatotoxin and nephrotoxin, is a product of HT degradation by ruminal microbes. Proanthocyanidins are considered to be non-toxic because they are not absorbed, but they are associated with lesions of the gut mucosa. Research on tannins in forage legumes has determined their effects on protein digestion and metabolism but more research on tannin structure in relation to digestion of specific proteins is needed. The widely accepted explanation for positive effects of PA on protein digestion and metabolism is that PA-protein complexes escape ruminal degradation and the protein is available in the lower tract. This proposed mechanism may be incorrect because PA also complex carbohydrates, endogenous proteins, and microbial products and the degradability of PA-protein complexes by ruminal microbes has not been adequately studied. Several alternative hypotheses (to escape protein) that explain the effect of PA on protein digestion and metabolism in ruminants are also consistent with experimental results on forage legumes. These include increased microbial protein synthesis, increased use of endogenous nitrogen in the rumen, and increased secretion of salivary glycoproteins. Research on manipulating the content and type of PA in forage legumes is justified because they are associated with non-bloating legumes, lower soluble non-protein nitrogen in silage, and improved efficiency of protein utilization. Research on the biosynthesis, molecular genetics, and cell biology of PA in forage legumes needs to be integrated with research on toxicology and nutrition.

Animal Feed↗

[Effects of tannins in animal nutrition].

Tannins are watersoluble plant polyphenols that precipitate proteins. According to their chemical structure they can be divided into condensed tannins and hydrolysable tannins. Altogether, tannins are reported to have various physiological effects like anti-irritant, antisecretolytic, antiphlogistic, antimicrobial and antiparasitic effects. Phytotherapeutically tannin-containing plants are used to treat nonspecific diarrhoea, inflammations of mouth and throat and slightly injured skins. Studies with ruminants have also demonstrated that the denaturing properties of tannins can possibly be used to improve protein supply to the small intestine. Besides anthelmintic effects of condensed tannins have been observed in sheep and goats. On the other hand high tannin concentrations resulted in reduced animal performances and health disorders as well in ruminants as in monogastriers. Further scientific research is therefore needed to clarify whether tannins can be effectivly used in livestock feeding, for example in prophylaxis of diarrhoea or parasitic control.

Animal Nutritional Physiological Phenomena↗

Inhibitory action of five tannins on growth induced by several gibberellins.

The following tannins, Chinese gallotannin, 1,2,3,4,6-pentagalloyl glucose, chebulinic acid, procyanidin dimers, and procyanidin trimers were tested and found to be antagonists of seven gibberellins (GAs). Each tannin inhibited the growth induced by any of the gibberellins GA(1), GA(3), GA(4), GA(7), GA(9), GA(13), and GA(14) in the dwarf pea assay. Endogenous growth was not affected. The highest ratio of tannin to gibberellin tested (1000:1 by weight) inhibited from 60 to 95% of the induced growth for all tannins and all gibberellins tested. The tannins were particularly inhibitory against GA(4) and GA(14) where a ratio of 10:1 (tannins: GA by weight) resulted in up to 85% growth reduction. Inhibition could be completely reversed by increasing the amount of gibberellin in all combinations studied. The procyanidin dimers and trimers were the first purified components of condensed tannins to be tested in this system and were potent inhibitors particularly against growth induced by GA(4) and GA(14). Inhibition by these compounds along with similar inhibition by previously tested hydrolyzable tannins demonstrates that the effect is general to tannins of all classes.

Journal Article↗

Hormetic versus toxic effects of vegetable tannin in a multitest study.

Tannin from mimosa trees (Acacia sp.) utilized in traditional leather tanning was tested for toxicity in sea urchin (Sphaerechinus granularis and Paracentrotus lividus) embryos and sperm, marine, and freshwater algae (Selenastrum capricornutum and Dunaliella tertiolecta), and Daphnia magna. Based on a two-step tanning procedure used in traditional tanneries, two mimosa tannin preparations, i.e., fresh tannin (FT) and used tannin (UT), were tested as suspensions. The early results in S. granularis embryos showed that UT exerted lower acute toxicity than FT, namely, 1 vs 100 mg/L, to obtain 100% mortality, respectively. Subsequent bioassays were conducted on fresh tannin water extracts (TWE) corresponding to nominal tannin concentrations ranging from 0.1 to 30 mg/L. Developmental toxicity, up to embryonic mortality was exerted by TWE at levels > 1 mg/L, S. granularis being more sensitive than P. lividus embryos/larvae. At the concentration of 0.1 mg/L, the frequencies of larval malformations were significantly lower than in controls. This positive stimulatory effect (currently termed as hormesis) was observed in extended numbers of culture replicates (up to 14) and was significant in the embryo cultures characterized by a relatively poor control quality (with < 70% viable larvae in controls), whereas this effect was not observed in good-quality cultures (with > or = 70% viable larvae in controls). Cytogenetic analysis of S. granularis embryos reared in FT or UT suspensions (1 mg/L to 1 g/L) showed mitotoxic effects (decrease in active mitoses per embryo) in FT-exposed, but not in UT-exposed embryos. Mitotic aberrations were significantly increased by 10 mg/L UT. Sperm fertilization success in both sea urchin species showed an increasing fertilization rate (FR) up to 0.3 mg/L TWE and a dose-related decrease in FR up to 30 mg/L. Again, the offspring of P. lividus sperm exposed to TWE (0.1 and 0.3 mg/L) showed a decrease in larval malformations compared to controls, whereas a dose-related increase in developmental defects was observed in the offspring of P. lividus sperm exposed to higher TWE levels (1 to 30 mg/L). Algal cell growth bioassays in two species (S. capricornutum and D. tertiolecta) also showed a maximum growth at TWE levels ranging from 0.3 to 3 mg/L and a subsequent decline up to 30 mg/L TWE. D. magna bioassays resulted in daphnid immobilization by TWE concentrations ranging from 100 to 300 mg/L. The results demonstrate that tannins utilized in traditional leather tanning industry may raise concern of environmental damage at relatively high concentrations, whereas low-level tannins may result in hormetic effects. The present study also points to the need for bioassay design that should rely on adequate criteria in control quality, allowing to detect both inhibitory and hormetic effects.

Acacia↗

Antihypertensive effects of tannins isolated from traditional Chinese herbs as non-specific inhibitors of angiontensin converting enzyme.

The tannins are natural polyphenols, able to precipitate water-soluble alkaloids and possess an inhibitory action on the angiotensin converting enzyme (ACE). We identified 18 polyphenolic compounds (tannins) from Chinese herbs and examined the in vitro effects of these tannins on ACE activity, including determination of the 50% inhibitory concentrations (IC50), specificity and mode of inhibition. We also assessed the in vivo inhibitory effect of the tannins on angiotensin I-induced blood pressure elevation in spontaneously hypertensive rats (SHR). Nine tannins with an IC50 <200 microM for ACE inhibitors were identified belonging to three tannin classes: caffeoylquinates, flavan-3-ols and gallotannins. In vitro, we found caffeoylquinates chelate the ACE zinc cofactor. Two of the flavan-3-ols: epigallocatechin-3-O-gallate and epigallocatechin-3-O-methylgallate, and one of gallotannin: 1, 2, 3, 4, 6-penta-O-galloyl-beta-D-glucose were non-specific inhibitors because also reduced the activity of trypsin and chymotrypsin. The ACE inhibition of 1, 2, 3, 4, 6-penta-O-galloyl-beta-D-glucose was also reduced after addition of bovine serum albumin, suggesting a non-specific mode of action. In vivo, 1,2,3,6-tetra-O-galloyl-beta-D-glucose and epigallocatechin-3-O-methylgallate had a strong dose-dependent hypotensive effect reducing the blood pressure significantly in the SHR with infusion of the angiotensin I. These findings indicate that some of the tannins isolated from herbs inhibit ACE activity non-specifically. The ACE inhibitory effect of these tannins may explain the hypotensive effects of some traditional Chinese herbs.

Angiotensin I↗