Analyses of PAHs (polycyclic aromatic hydrocarbons) and mutagenicity in raw and chlorinated water

Both raw water and chlorinated drinking water samples were collected from and the Liu‐Du water treatment plant in northern Taiwan from October 1990 to April 1992. The polycyclic aromatic hydrocarbons (PAHs) and mutagenicity in these water samples were analyzed by GC/MS and Ames test. The Mutagenicity/DMSO (Dimethyl Sulfoxide) ratio in S. typhimurium TA98 and TA100 with or without S9 mixture increased, even higher than 2, following the sequence of unit process. It was observed that the mutagenicity with TA98 (S9+) was highly related to most of PAHs in the raw water; while the mutagenicity with TA98 (S9+) was only correlated with DbA and BghiPr in the treated water. It could be expected that the mutagenicity level was controlled by other predominant components after the raw water was treated, for example, the chlorination process.     

Multiple-time contamination of benzo(a)pyrene in soils inhibits soil enzymatic activities

Benzo[a]pyrene (B[a]P) is accumulating in soils in a low-dose cumulative manner. The objectives of this study were to investigate the effects of B[a]P on the extractable and available fractions of B[a]P and on soil enzymatic activity using multiple-time superimposed and one-time contamination approaches. Results showed that the contents of B[a]P rapidly decreased in the first 14?d and later decreased slowly from 14 to 56?d in both one-time and multiple-time contamination tests. The contents of B[a]P in the multiple-time contamination test were lower than those in the one-time test. Soil urease, sucrase and dehydrogenase activities were rapidly inhibited in the early stage (14?d) and stimulated during the rest of the incubation, and soil dehydrogenase activity was more sensitive to B[a]P contamination than the other enzymes. High concentrations of B[a]P in soil led to greater inhibition of enzymatic activity than that at low concentrations in the early period of culture. Soil enzyme activities were weakly inhibited in multiple-time compared with in one-time contamination tests and were lower in the subsurface layer than in the surface layer. Our results revealed that the multiple-time superimposed approach might be better than one-time contamination for evaluating B[a]P risk in soil.     

Mutagenicity of diesel engine exhaust in the Ames / Salmonella assay using a direct exposure method

The aim of this study was to investigate the potential mutagenic activity of diesel engine exhaust in the Ames/Salmonella assay using a direct aerosol exposure system. So, TA 98 and TA 100 strains, with or without added S9 mix, were exposed to diesel emissions after varying degrees of filtration. Variants of these two strains, deficient in nitroreductase (TA 98NR and TA 100NR) or over-expressing O-Acetyl Transferase (YG 1024 and YG 1029), were also exposed to total (unfiltered) diesel exhaust to highlight the putative mutagenicity of any nitro-PAHs present in these emissions. Mutagenic activity of the diesel exhaust was demonstrated on Salmonella typhimurium, strains TA 100 and variants TA 100 NR and YG1029. The use of a particle filter did not modify the genotoxicity of the diesel emissions, indicating a major contribution of the gas phase to the mutagenicity of these diesel emissions. The prominent role of the particulate-associated nitro-polycyclic aromatic hydrocarbons (nitro-PAHs) claimed by some authors working on diesel exhaust organic extracts was not confirmed by our results with native diesel exhaust exposure. Our results show that the gas phase is potentially more mutagenic than the particles alone.     

Degradation of Benzo[a]Pyrene in Soil with Arbuscular Mycorrhizal Alfalfa

Mycorrhizal and non-mycorrhizal alfalfa (Medicago sativa) was grown in pots containing soil artificially contaminated with various levels of benzo[a]pyrene (B[a]P)(0, 1, 10 and 100 mg kg^–1). Soil and plants were sampled after 30, 40, 50, 60 and 90 days and compared with unlanted pots. The percentage of mycorrhizal root length colonized by Glomus caledoniun was not significantly affected by the addition of B[a]P up to 10 mg kg^–1 but was significantly lower at 100 mg kg^–1B[a]P compared with low concentrations (p < 0.05). There was no difference in soil polyphenol oxidase and dehydrogenase activity among the controls and applications of 1 and 10 mg kg^–1 of B[a]P. However, enzyme activities were significantly higher at 100 mg kg^–1B[a]P compared with the other three treatments, and there was no mycorrhizal effect. Over a period of 90 days the concentration of B[a]P in soil in which alfalfa was grown was significantly lower than in unplanted soil (p < 0.05). Degradation rates of B[a]P added at 1, 10 and 100 mg kg^–1 without G. caledonium were 76, 78 and 53%, and with mycorrhizal inoculation were 86, 87 and 57%. The degradation rate in unplanted soil was significantly lower than in planted soil, and was significantly higher in medium- and low-B[a]P treatments than in the high B[a]P concentration tested. There is a possibility of enhancement phytoremediation of PAHs in rhizosphere soil with arbuscular mycorrhizal fungi.     

Antibacterial and antimutagenic activitiy of extracts and essential oil from (Tunisian) Pistacia lentiscus

Aqueous and flavonoid-enriched extract as well as essential oil (EO) obtained from leaves of Pistacia lentiscus were assessed for antibacterial and antimutagenic activities. Antibacterial activity of different extracts and EO were evaluated against six bacterial strains. A marked inhibitory effect was observed against Salmonella typhimurium, whereas lower activity was observed against Staphylococcus aureus, Pseudomonas aeruginosa, and Salmonella enteritidis. EO showed significant inhibitory effects against Salmonella typhimurium, Salmonella enteritidis and Staphylococcus aureus. The antimutagenic activity of the different extracts against Aflatoxin B₁ (AFB₁) and sodium azide was demonstrated with the Salmonella typhymurium assay. The number of revertants per plate decreased significantly when the plant extracts were added to the assay system using Salmonella typhimurium TA100, TA98 and TA1535.     

Chemopreventive effects of ellagic acid against genotoxicity induced by benzo(a)pyrene

Benzo(a)pyrene (B(a)P) is a commonly used indicator for polycyclic aromatic hydrocarbons (PAHs) contamination. The use of certain phytochemicals and some polyphenolic compounds proved to be effective in blocking PAH-induced effects. The aim of the present study was to examine the possible inhibitory effects of ellagic acid (EA), present in berries and nuts, against B(a)P-mediated toxicity using in vitro and in vivo test systems. In vitro method included the Ames test, using Salmonella typhimurium tester strain TA100. In vivo experiments were based on mouse bone marrow micronucleus (MN) assay and immunoblotting. EA produced a decrease in the number of revertant colonies against B(a)P in the Ames assay. Further, there was a reduction in the mean number of micronucleated polychromatic erythrocytes in the presence of EA in the co-treatment protocol of the MN test. Western blotting results showed downregulation of CYP450 isoform CYP1A1 which prevented bioactivation of B(a)P. Thus, the present study demonstrates antimutagenic role played by EA against the mutagenic activity of B(a)P in both in vitro and in vivo systems.     

Genotoxicity,antifungal and antibacterial activity of newly synthesized N-(3-phthalidyl)amines and o-benzoyl benzamide derivatives

A number of newly synthesized phthalidylamines and o-benzoylbenzamide derivatives were evaluated for some biological activities. Synthesis was established by condensation of 3-acetoxyphthalide 1 with morpholine, piperidine, N,N-diisobutyl-N,N-dibenzylamines and piperazine, which afforded N-(3-phthalidyl)amines 3a–d, and 4 respectively, while with N,N-diisopropylamine, o-formyl-N,N-diisopropyl benzamide 5a is formed exclusively. On the other hand, the reaction of 3-acetoxy-3-phenylphthalide 2 with secondary amines afforded o-benzoylbenzamide derivatives 5b–c, 6 in a high yield. The structure of the reaction products was established from their spectral data. These products were screened for antifungal, antibacterial and genotoxic effect. It was found that all tested compounds have antifungal activity. Compounds 1, 2, 3d and 5b were found to be active against Escherichia coli, Bacillus subtilis and Staphylococcus aureus. Genotoxic effects using Ames test showed that Compounds 1 and 2 have a weak base-pair substitution mutagenicity while a clear base-pair substitution mutagenic activity was shown by 3a using TA100-strain of Salmonella typhimurium. Compound 4 showed a frameshift mutgenicity while a weak oxidative mutagenic action was revealed by 6. No change on the mutagenicity of the tested chemicals was observed after using the S9 metabolic activation system.     

Induction of gene mutations in salmonella and Drosophila by soluble Cr(Vi) compounds: Synergistic effects of nitrilotriacetic acid

The interaction between NTA and soluble Cr(VI) (K₂Cr₂O₇) was studied by the Ames test on S. typhimurium and the sex‐linked recessive lethal test on D. melanogaster. In both systems a synergistic effect of NTA on Cr(VI) mutagenicity took place at sub‐toxic doses of Cr(VI). The synergism could depend on the action of NTA on intracellular Cr(VI) reduction, as more Cr(VI) was reduced in vitro to Cr(III) by Salmonella and Drosophila protein extracts in the presence of NTA. A similar enhancement of soluble Cr(VI) mutagenicity was produced by low doses of EDTA.     

Protection of Genetic Diversity and Maintenance of Connectivity among Reef Corals within Marine Protected Areas

Abstract: High‐latitude coral reefs (HLRs) are potentially vulnerable marine ecosystems facing well‐documented threats to tropical reefs and exposure to suboptimal temperatures and insolation. In addition, because of their geographic isolation, HLRs may have poor or erratic larval connections to tropical reefs and a reduced genetic diversity and capacity to respond to environmental change. On Australia's east coast, a system of marine protected areas (MPAs) has been established with the aim of conserving HLRs in part by providing sources of colonizing larvae. To examine the effectiveness of existing MPAs as networks for dispersal, we compared genetic diversity within and among the HLRs in MPAs and between these HLRs and tropical reefs on the southern Great Barrier Reef (GBR). The 2 coral species best represented on Australian HLRs (the brooding Pocillopora damicornis and the broadcast‐spawning Goniastrea australensis) exhibited sharply contrasting patterns of diversity and connectedness. For P. damicornis, the 8‐locus genetic and genotypic diversity declined dramatically with increasing latitude (N_a= 3.6–1.2, H_e= 0.3–0.03, N_g:N = 0.87–0.06), although population structure was consistent with recruitment derived largely from sexual reproduction (G_o:G_e= 1.28–0.55). Genetic differentiation was high among the HLRs (F_ST[SD]= 0.32 [0.08], p < 0.05) and between the GBR and the HLRs (F_ST= 0.24 [0.06], p < 0.05), which indicates these temperate populations are effectively closed. In contrast for G. australensis, 9‐locus genetic diversity was more consistent across reefs (N_a= 4.2–3.9, H_e= 0.3–0.26, N_g:N = 1–0.61), and there was no differentiation among regions (F_ST= 0.00 [0.004], p > 0.05), which implies the HLRs and the southern GBR are strongly interconnected. Our results demonstrate that although the current MPAs appear to capture most of the genetic diversity present within the HLR systems for these 2 species, their sharply contrasting patterns of connectivity indicate some taxa, such as P. damicornis, will be more vulnerable than others, and this disparity will provide challenges for future management.     

Parameters of light-dependent photosynthesis for phytoplankton size fractions in temperate estuarine and coastal environments

Photosynthetic parameters for netplankton (>22 m) and nanoplankton (<22 m) varied over similar ranges but exhibited different seasonal and geographic patterns of variation. Nanoplankton a was relatively constant (0.06 mg C [mg Chl · h]^-1 [E m^-2 s^-1]^-1), but P _m ^B (mg C [mg Chl · d]^-1) was an exponential function of temperature independent of nutrient concentration and vertical stability in the euphotic zone. The temperature function gives a P _m ^B of 24 at 25°C for nanoplankton growing in an estuarine environment characterized by high nutrient concentrations and a shallow, stratified euphotic zone. Variations in netplankton a and P _m ^B were less predictable and were not correlated with temperature, nutrients or vertical stability. Chain forming diatoms with small cells were able to achieve high (0.10 to 0.15) and P _m ^B (20 to 24) that were 3 to 5 times higher than large-celled diatoms and dinoflagellates were able to achieve.     

Acid-base responses to lethal aquatic hypercapnia in three marine fishes

Ocean sequestration of CO₂ is proposed as a possible measure to mitigate environmental changes due to the increasing atmospheric concentration of the gas. However, toxic effects of CO₂ on marine organisms are poorly understood. We therefore studied acid–base responses and mortality during exposure to fatal levels of CO₂ in three marine fishes (Japanese flounder, Paralichthys olivaceus; yellowtail, Seriola quinqueradiata; and starspotted dogfish, Mustelus manazo). The teleosts died during exposure to seawater equilibrated with a gas mixture containing 5% CO₂ (water PCO₂ 4.95 kPa); 100% mortality occurred within 8 h for yellowtail and within 48 h for flounder. Only 20% mortality was recorded at 72 h for the dogfish during exposure to 7% CO₂ (water PCO₂ 6.96 kPa). Arterial pH (pH_a) initially decreased, but completely recovered within 1–24 h for the teleosts at 1% and 3%, although the recovery was slower and complete only at 1% (water PCO₂ 0.99 kPa) for the dogfish. During exposure to 5%, the flounder died after the pH_a had been completely restored, suggesting that the mortality was not due to plasma acidosis. During exposure to 1% hypercapnia, plasma [Cl^–] appeared to be the main counter ion to balance increases in plasma [HCO₃^-]. There was a 1:1 stoichiometry for the rise in [HCO₃^-] and the fall in [Cl^–] for the teleosts, whereas the ratio was 1:0.4 for the dogfish at 1% CO₂. At the higher levels of hypercapnia, the rise in [HCO₃^-] consistently exceeded the fall in [Cl^–], and plasma [Na⁺] significantly increased.These results do not agree with the generally accepted model for acid–base regulation in marine fish in which Na⁺/H⁺ exchangers are assumed to play a predominant role, and indicate that an acid–base regulatory mechanism differs between teleost and elasmobranch fishes, as well as the intensity of acidic stress.Communicated by T. Ikeda, Hakodate     

Evaluation of chlorophyll fluorescence, in vivo spectrophotometric pigment absorption and ion leakage as biomarkers of UV-B exposure in marine macroalgae

The photosynthetic fluorescence ratio F_v:F_m, in vivo absorption spectra and ion leakage were evaluated as biomarkers of ambient and elevated UV-B (280 to 320 nm) exposure of the intertidal alga Enteromorpha intestinalis (Chlorophyta) and the sublittoral alga Palmaria palmata (Rhodophyta). Measurements of thallus growth were also used to assess adverse biological effects. Ambient and elevated UV-B significantly inhibited photosynthesis in both species. It was shown that the F_v:F_m ratio is a sensitive, non-specific general biomarker of UV-B exposure in both species. Moreover, the in vivo absorption of what was tentatively identified as chlorophylls a and b as well as phycoerythrin and/or carotenoids, phycoerythrobilin and phycocyanin decreased in a dose-response dependent manner and was associated with a decrease in growth rate in P. palmata. The intertidal alga E. intestinalis showed a greater degree of tolerance to UV-B exposure. These results indicate that changes in the F_v:F_m ratio together with reductions in in vivo pigment absorption could provide an early quantitative warning of the detrimental effects of UV-B in marine macroalgae. Received: 16 May 1997 / Accepted: 16 July 1997     

Gingko biloba extract and cobalt porphyrin additive to remove harmful components from cigarette smoke and reduce its toxicity

Cigarette filters were modified with a combination of gingko biloba extract and cobalt porphyrin (CGC) to remove harmful components from the cigarette smoke and reduce its toxicity. Smoke analysis results indicated that CGC eliminated up to 32% of benzo[a]pyrene (B[a]P), 52% of N-nitrosonornicotine (NNN), 46% of N-nitrosoanabasine (NAB), 35% of 4-(methylnitrosamine)-1-(3-pyridyl)-1-butanone (NNK), 31% of N-nitrosoanatabine (NAT), 30% of gas-phase free radicals, and 33% of solid-phase free radicals. Biological experiments, including the Ames test, neutral red cytotoxicity assay and chronic toxicity, were conducted for both CGC cigarettes and control cigarettes. Results showed that the toxicity of the CGC cigarettes was lower than those of the control cigarettes. The mechanism by which the CGC components could remove harmful components from cigarette smoke is discussed.     

Age-specific responses to elevated salinity in the coastal marsh plant black needlerush (Juncus roemerianus Scheele) as determined through polyphasic chlorophyll a fluorescence transients (OJIP)

This study employed polyphasic chlorophyll a fluorescence transients (OJIP), a non-invasive marker of environmental stress in plants, to evaluate salt tolerance in three different Juncus roemerianus age classifications (6-, 24-, and 60-months). Following exposure to elevated salts (30 psu), the younger plants sustained growth, which was comparable to freshwater controls. While older (60-month) plants receiving only freshwater also grew over the 8-week study, the older salt-treated plants did not increase in size. Similarly, there were significant declines in variable chlorophyll a fluorescence parameters (F _v/F _m and F _v/F _o), electron transport flux per reaction center (ET_o/RC), and photosystem II performance index (PI_ABS) for 60-month J. roemerianus following salt treatment. These responses were not evident in the two younger salt-treated age classifications. Our results suggest that older J. roemerianus are less tolerant to rapid and sudden increases in salinity relative to younger plants and that this age-specific response may help explain observed discrepancies in salt tolerance in J. roemerianus.     

Nitropyridoindoles—Formation,characterization and mutagenicity

Nitration of pyridoindoles (β‐carbolines) was carried out using different nitration methods and structural assignments of these nitro compounds were made using different spectroscopic techniques. The mutagenic activity of these nitro derivatives was studied using Salmonella typhimurium strains TA 98 NR and TA 100 NR in the absence as well as in the presence of S‐9 mix. Only trinitro derivatives were found to be highly mutagenic to strain TA 98 NR.     

DCMU-enhanced fluorescence as an index of photosynthetic activity in phytoplankton

Tests have been carried out, both on phytoplankton cultures and in the field, on the relation between the ratio of 3-(3,4-dichlorophenyl)-1,1-dimethyl urea (DCMU)-enhanced to normal fluorescence (F _D /F _N ) and the specific photosynthetic rate (P/B). In all cases observed, significant linear correlations were found between these two values. Differences in this relation were observed according to species and physiological conditions. Nutrient stress that occurs in batch cultures decreases both F _D /F _N and P/B, while lowered light intensity has a different effect on both, increasing F _D /F _N and decreasing P/B. This is interpreted as indicating that F _D /F _N is an index of photosynthetic efficiency at different light levels and an indicator of the specific photosynthetic rate (P/B) when physiological conditions vary at a given light intensity. The practical use of DCMU-enhanced fluorometry to estimate instantaneous P/B values in the field is discussed, stressing the frequent calibrations needed, as in all in situ fluorometry studies.     

Quantum yield,relative specific absorption and fluorescence in nitrogen-limited Chaetoceros gracilis

Decreases in cell-nitrogen quota resulted in changes in the carbon-based quantum yield of photosynthesis, the chlorophyll a-specific absorption coefficient, and in vivo fluorescence in the marine diatom Chaetoceros gracilis in laboratory experiments performed in 1983 and 1984. The three parameters were independently determined for the two spectral regions dominated by either chlorophyll a or fucoxanthin absorption. As cell-nitrogen quota decreased, the quantum yield for both pigments decreased; the specific absorption coefficient for chlorophyll a and the in vivo chlorophyll a fluorescence excited by each pigment increased. The observed increase in the in vivo fluorescence per chlorophyll a could be partially attributed to the increased specific absorption coefficient for chlorophyll a; the remainder of the fluorescence increase was related to a decline in photosystem activity. Energy transfer efficiency between light-harvesting pigments appeared to be maintained as cell-nitrogen quota decreased. The decrease in a fluorescence index [(F _DCMU-F _O)/F _DCMU] with nitrogen starvation suggested a decrease in Photosystem II activity. These results imply that decreases in reaction center and/or electron-transport system activity were responsible for the decline in rates of photosynthesis under conditions of notrogen deficiency.     

Fluorescence studies of dye displacement from DNA by chromium(III) complexes: Evidence for cation induced DNA condensations

The interactions of 10 different chromium(III) complexes with isolated calf thymus DNA have been analysed by studying the electronic and fluoresence spectra of intercalated ethidiumbromide. Triply charged cationic complexes including: [Cr(urea)₆]Cl₃.3H₂O, [Cr(1,10‐phenanthroline)₃](ClO₄)₃.2H₂O, [Cr(2,2'‐bipyridyl)₃] (ClO₄)₃.2H₂O, [Cr(ethylendiamine)₃]Cl₃.3.5H₂O and [Cr(NH₃)₆](NO₃)₃ displaced the dye from DNA. Similar effects were observed in experiments using the non‐intercalating dye bisbenzimidazole ("Hoechst 33258"). However, singly charged cationic, anionic and uncharged chromium(III) complexes such as: cis‐[Cr(1,10‐phenanthroline)₂Cl₂]Cl.2H₂O, cis‐[Cr(2,2'‐bipyridyl)₂Cl₂]Cl.2H₂O, [Cr(glutathione)₂]Na₂, [Cr(cysteine)₂]Na.2H₂O and [Cr(glycine)₃] were unable to displace both ethidiumbromide and bisbenzimidazole from DNA. There was no evidence for the formation of co‐ordinate bonds between chromium(III) and DNA for any of the above complexes. The charge and type of ligand are important in controlling the interaction of chromium(III) with isolated DNA in vitro. Our findings indicate that the outer sphere interaction of a chromium(III) complex with DNA is weak and unlikely to be the mechanism by which chromate causes DNA impairments in vivo and in vitro.     

Availability of ammonium influences photosynthesis and the accumulation of mycosporine-like amino acids in two<Emphasis Type="Italic"> Porphyra</Emphasis> species (Bangiales,Rhodophyta)

The effect of ammonium concentration on photosynthetic activity estimated as in vivo chlorophyll fluorescence, i.e. maximal quantum yield (F_v/F_m) and electron transport rate (ETR) and on the accumulation of mycosporine-like amino acids (MAAs), chlorophyll a (chl a), biliproteins (BP) and soluble proteins (SP) in the red algae Porphyra leucosticta Thuret in Le Jolis collected from Lagos (Málaga, Spain) and Porphyra umbilicalis (Linnaeus) J. Agardh from Helgoland (Germany) was evaluated. Discs of both species were incubated with three ammonium concentrations (0, 100 and 300 µM) under artificial PAR and UV radiation for 7 days. Photosynthetic activity decreased under the culture conditions due to UV radiation and ammonium availability. The decrease of both F_v/F_m and maximal ETR was related to ammonium supply, i.e. the lowest decrease occurred in algae growing with the highest concentration of ammonium. In both species, after 7 days of culture, the content of chl a, BP and SP was higher under 300 µM than that under 0 and 100 µM ammonium. In both species, the content of MAAs was increased under 300 µM ammonium compared to the initial value, whereas a decrease under 0 and 100 µM ammonium was observed only in P. leucosticta. The content of MAAs in P. umbilicalis did not present significant differences compared to the initial value, probably because of the high initial content of MAAs. In both Porphyra species, four MAAs were identified: shinorine, porphyra-334, palythine and asterina-330. However, P. leucosticta modified its MAA pattern during the incubation time, reaching the same percentages found for P. umbilicalis, which did not show any change during the experimental period. P. leucosticta exhibited a decrease in BP/SP and BP/chl a ratios through the incubation time and an increase in MAAs/BP. The ratio MAAs/chl a did not show any variation with time or treatment, as was also true for all ratios in P. umbilicalis. In summary, ammonium supply diminished the decrease of F_v/F_m, increased the content of photosynthetic pigments (chlorophyll and biliprotein) and soluble protein, and stimulated of the accumulation of MAAs in the red algae P. leucosticta and P. umbilicalis.Communicated by O. Kinne, Oldendorf/Luhe     

Evaluation of the antimutagenic and antiradical potentials of extracts from the tubers of (Tunisian) Cyperus rotundus

The mutagenic potential of aqueous, total oligomers flavonoids (TOF), ethyl acetate and methanol extracts from tubers of Cyperus rotundus L. was assessed using Ames Salmonella tester strains TA98 and TA100, and SOS chromotest strain Escherichia coli PQ37 with and without metabolic activation (S9). None of the different extracts produced a mutagenic effect. Likewise, the antimutagenicity of the same extracts was tested using the “Ames test” and the “SOS chromotest”. Our results showed that C. rotundus extracts possess antimutagenic effects against S. typhimurium TA98 and TA100 strains towards the mutagen aflatoxin B1 (AFB1), similar to E. coli PQ37 strain against AFB1 and Nifuroxazide mutagens using the SOS chromotest assay. A free radical scavenging test was used in order to explore the antioxidant capacity of the extracts obtained from the tubers of C. rotundus. TOF, ethyl acetate and methanol extracts showed an important free radical scavenging activity towards the 1,1-diphenyl-2-picrylhydrazine (DPPH) free radical. These extracts showed an IC50 value of respectively 5, 20 and 65?µg?mL^?1. The beneficial effects of TOF, ethyl acetate and methanol extracts of C. rotundus were assessed by antioxidant and antimutagenic activities.