Evaluation of genotoxicity of medicinal plant extracts by the comet and VITOTOX tests.

We report the results of our genotoxic evaluation of extracts from three medicinal plants Acacia nilotica, Juglans regia, and Terminalia chebula and the herbal drug Triphala employing the VITOTOX and comet tests.These tests detect DNA damage in prokaryotic and eukaryotic test systems, respectively. In the VITOTOX test, none of the extracts were identified as genotoxic. In the comet assay, extracts of Acacia nilotica showed statistically significant DNA damage only in a concentration of 2500 ppm (highest tested dose), whereas extracts from Juglans regia showed significant damage in concentrations above 250 ppm and more. Extracts from Terminalia chebula and Tripahala significantly increased DNA damage in a concentration above 500 ppm. This is not considered contradictory, because DNA damage in the alkaline comet assay may not be permanent and hence may not necessarily result in mutations. All the extracts were previously found in the Ames assay to have potent antimutagenic effects against the direct acting mutagens NPD, sodium azide, and the S9-dependent mutagen 2-AF. The results of the previous study using the Ames assay are in conformity with those of the VITOTOX test. It was found that the extracts were safe in concentrations of up to 1000 microg/0.1 mL and 2500 microg/0.1 mL. A literature survey also showed that plant extracts can be mutagenic as well as antimutagenic depending on the test system used. This indicates that a battery of assays is needed before any conclusion can be reached.

Final report of the safety assessment of Acacia catechu gum, Acacia concinna fruit extract, Acacia dealbata leaf extract, Acacia dealbata leaf wax, Acacia decurrens extract, Acacia farnesiana extract, Acacia farnesiana flower wax, Acacia farnesiana gum, Acacia senegal extract, Acacia senegal gum, and Acacia senegal gum extract.

These ingredients are derived from various species of the acacia plant. Only material derived from Acacia senegal are in current use according to industry data. The concentration at which these ingredients are reported to be used ranges from 9% in mascara to 0.0001% in tonics, dressings, and other hair-grooming aids. Gum arabic is a technical name for Acacia Senegal Gum. Gum arabic is comprised of various sugars and glucuronic acid residues in a long chain of galactosyl units with branched oligosaccharides. Gum arabic is generally recognized as safe as a direct food additive. Little information is available to characterize the extracts of other Acacia plant parts or material from other species. Acacia Concinna Fruit Extract was generally described as containing saponins, alkaloids, and malic acid with parabens and potassium sorbate added as preservatives. Cosmetic ingredient functions have been reported for Acacia Decurrens Extract (astringent; skin-conditioning agent--occlusive) and Acacia Farnesiana Extract (astringent), but not for the other Acacias included in this review. Toxicity data on gum arabic indicates little or no acute, short-term, or subchronic toxicity. Gum arabic is negative in several genotoxicity assays, is not a reproductive or developmental toxin, and is not carcinogenic when given intraperitoneally or orally. Clinical testing indicated some evidence of skin sensitization with gum arabic. The extensive safety test data on gum arabic supports the safety of Acacia Senegal Gum and Acacia Senegal Gum Extract, and it was concluded that these two ingredients are safe as used in cosmetic formulations. It was not possible, however, to relate the data on gum arabic to the crude Acacias and their extracts from species other than Acacia senegal. Therefore, the available data were considered insufficient to support the safety of Acacia Catechu Gum, Acacia Concinna Fruit Extract, Acacia Dealbata Leaf Extract, Acacia Dealbata Leaf Wax, Acacia Decurrens Extract, Acacia Farnesiana Extract, Acacia Farnesiana Flower Wax, Acacia Farnesiana Gum, and Acacia Senegal Extract in cosmetic products. The additional data needed to complete the safety assessment for these ingredients include (1) concentration of use; (2) identify the specific chemical constituents, and clarify the relationship between crude Acacias and their extracts and the Acacias and their extracts that are used as cosmetic ingredients; (3) data on contaminants, particularly relating to the presence of pesticide residues, and a determination of whether Acacia melanoxylon is used in cosmetics and whether acamelin (a quinone) and melacacidin (a flavin) are present in the Acacias that are being used; (4) skin sensitization study (i.e., dose response to be determined); (5) contact urticaria study at use concentration; and (6) ultraviolet (UV) absorption spectrum; if there is significant absorbance in the UVA or UVB range, then a photosensitization study may be needed. It was also noted that other data may be needed after clarification of the chemical constituents of the Acacia-derived ingredients.

Use of community genome arrays (CGAs) to assess the effects of Acacia angustissima on rumen ecology.

This research developed a community genome array (CGA) to assess the effects of Acacia angustissima on rumen microbiology. A. angustissima produces non-protein amino acids as well as tannins, which may be toxic to animals, and CGA was used to assess the effects of this plant on the ecology of the rumen. CGAs were developed using a 7.5 cmx2.5 cm nylon membrane format that included up to 96 bacterial genomes. It was possible to separately hybridize large numbers of membranes at once using this mini-membrane format. Pair-wise cross-hybridization experiments were conducted to determine the degree of cross-hybridization between strains; cross-hybridization occurred between strains of the same species, but little cross-reactivity was observed among different species. CGAs were successfully used to survey the microbial communities of animals consuming an A. angustissima containing diet but quantification was not precise. To properly quantify and validate the CGA, Fibrobacter and Ruminococcus populations were independently assessed using 16S rDNA probes to extracted rRNA. The CGA detected an increase in these populations as acacia increased in the diet, which was confirmed by rRNA analysis. There was a great deal of variation among strains of the same species in how they responded to A. angustissima. However, in general Selenomonas strains tended to be resistant to the tannins in the acacia while Butyrivibrio fibrisolvens was sensitive. On the other hand some species, like streptococci, varied. Streptococcus bovis-like strains were sensitive to an increase in acacia in the diet while Streptococcus gallolyticus-like strains were resistant. Strep. gallolyticus has independently been shown to be resistant to tannins. It is concluded that there is significant variation in tannin resistance between strains of the same species. This implies that there are specific molecular mechanisms at play that are independent of the phylogenetic position of the organism.

Effect of feeding Acacia abyssinica and of its extracts given by different routes on rats.

The effect of a diet consisting of 2% and 10% of Acacia abyssinica bark on Wistar rats treated for 6 weeks was examined. A 2% A. abyssinica diet was not toxic to rats. Impairment of growth and hepatonephropathy were observed in rats on a 10% A. abyssinica diet. By whatever route it was administered, either intraperitoneally (i.p.) or orally (p.o.), the ethyl acetate extract in daily doses of 500 mg/kg body weight was the most toxic and lethal to rats and caused hepatonephropathy, widespread hemorrhage and congestion and fibrinous peritonitis following i.p. administration. The aqueous and ethanol extracts i n similar doses to ethyl acetate extract were only lethal to rats when given via the i.p. route. Lesions were accompanied by anemia, leukopenia and alterations in serum AST activity and concentrations of urea, total protein and albumin.

A study on the toxicology of Acacia nilotica.

The potential toxicity of Acacia nilotica was investigated in rats maintained on 2% and 8% acacia diet for 2 and 4 weeks. A significant reduction in body weight in all acacia-fed groups and a significant decrease in the levels of hemoglobin, serum total protein and total cholesterol in animals fed 8% acacia diet for up to 4 weeks were observed. These effects were, however, reversed one week after treatment termination. No significant changes in serum parameters of hepatic and renal functions, fasting glucose and triglycerides were observed. Further, no deaths among treated animals and no significant histopathological changes in liver sections were noted. It is concluded that A. nilotica, at 2% and 8% levels, has a low toxicity potential.