Mutagenic Index Research Articles

Heterocyclic phytometabolites formononetin and arbutin prevent in vitro oxidative and alkylation-induced mutagenicity

Phenolic phytometabolites are promising bioactive compounds for management of genomic instability related diseases. Formononetin (FMN) and arbutin (ARB) are found in several plant sources. Our goal was to investigate the safety and efficacy of FMN and ARB using in vitro both standardized and alternative toxicogenetic methods. FMN and ARB were evaluated through the OECD’S guidelines No. 471 (Bacterial Reverse Mutation Test –Salmonella/microsome) and No. 487 (In vitro Mammalian Micronucleus Test – CBMN assay), accordingly to the mentioned recommendations. Also, antimutagenicity of FMN and ARB was assessed in S. Typhimurium strains TA98, TA100 and TA1535, following pre-, co- and post- treatment protocols. Liver human lineages HepG2 and F C3H were assayed for cytotoxicity after exposure to FMN and ARB (24, 48 and 72 h) using in vitro WST-1 test. ARB showed no mutagenicity in the Salmonella/microsome test under both metabolic conditions (in presence or absence of 4 % S9 mix), but FMN was cytotoxic to the TA97 and TA100 strains after metabolic activation. Under this same condition, FMN induced an increase in the mutagenic index of strain TA1535 at two of the highest tested concentrations. Even so, ARB and FMN exhibited protection against the induced alkylation of DNA in multiple action modes. In the antimutagenicity assay, FMN reached the maximum of 80 % of oxidative-provoked mutagenicity reduction in TA98 strain in co-treatment with known mutagen, besides 69 % of reduction in TA100 in the same exposure condition. ARB showed up to reduce induced mutagenicity in strains TA100 and TA1535, reaching percentages from 55 % to 100 % of antimutagenicity in all of the tested exposure models against alkylating agent. In the CBMN assay, no increase in micronuclei formation was observed. The results suggest that FMN and ARB prevent DNA from mutation using multi-targeted antimutagenic roles. Finally, our data suggests that FMN and ARB are not genotoxic and presented encouraging antimutagenicity action in vitro, being promising compounds for use in genomic instability-related diseases therapeutics.

Cytotoxicity, genotoxicity, and mutagenicity of the active pharmaceutical ingredient nevirapine and a nevirapine-based drug on the plant species Allium cepa

The toxicity of the active pharmaceutical ingredient and nevirapine-based drug at analytical concentrations was evaluated under laboratory conditions, using Allium cepa seeds as a model. The germination index of the negative control was 86.8 ± 2.1. The concentrations of 6.42 and 9.54 mg/L of the active pharmaceutical ingredient and 11.20 mg/L of the nevirapine-based drug showed a statistically higher germination index than the negative control. We found that the root growth of the negative control was 1.7 ± 0.6 mm and that the root growth was statistically lower than the negative control at concentrations of 9.54 and 17.73 mg/L of active pharmaceutical ingredient and 5.48, 11.20, and 17.68 mg/L of the drug. The mitotic index of negative control and methyl methanesulfonate were 7.4 ± 2.7 and 12.8 ± 4.5, respectively. At a concentration of 17.68 mg/L of the nevirapine-based drug, the mitotic index of 12.7 ± 2.7 was statistically higher than the negative control and like the methyl methanesulfonate, which indicated that nevirapine was cytotoxic at this concentration. At all concentrations tested the chromosome abnormality indices were statistically higher than negative control, suggesting that nevirapine was genotoxic. The mutagenicity index of the negative control was 0.2 ± 0.3. At concentrations of 6.42, 9.54, and 17.73 mg/L of the active pharmaceutical ingredient and 17.68 mg/L of the nevirapine-based drug, the mutagenicity index was statistically higher than the negative control, indicating that nevirapine was mutagenic for A. cespa at these concentrations. The chromosomal adhesion was the most frequent chromosomal aberration in the groups exposed to nevirapine, suggesting that it has an aneugenic effect on the A. cepa species.

Genotoxicity evaluation of two derived products from Allium extracts: s-propylmercaptocysteine and s-propyl mercaptoglutathione

Propyl-propane-thiosulfonate (PTSO) is one of the main organosulfur compounds present in Allium essential oils with a widely documented biological activity. For this reason, it could be used as a food and feed additive in the agri-food industry. A genotoxicity evaluation of substances and their metabolites present in food is necessary to guarantee the consumer's health following the recommendations of the European Food Safety Authority (EFSA). To evaluate the toxicological profile of derivatives of PTSO, the cytotoxicity, an Ames test, a micronucleus test and the comet assay were performed. Results showed that non-cytotoxic effects were observed in Caco-2 exposed to s-propyl mercaptocysteine (CSSP) and s-propyl mercaptoglutathione (GSSP) (0–450 μM). The mutagenicity index remained in the range of 0.6–1.4 for both compounds, showing no mutagenic effects for the concentrations of 5000–312.5 μg GSSP/plate and 250–15.63 μg CSSP/plate. Moreover, the % binucleated cells with micronuclei were 1.3–2.2 and 1.6–2.7 for GSSP and GSSP, respectively. For comet assays there was no DNA-genotoxic or oxidative damage in a concentration range of 112.5–450 μM. Therefore, we can conclude that these compounds are not genotoxic at the conditions tested. These results support that the presence of CSSP and GSSP in the food/feed is not of concern, although further studies are needed to complete their safety profile.

Reduced genotoxicity of lignin-derivable replacements to bisphenol A studied using in silico, in vitro, and in vivo methods

Bisguaiacols, lignin‐derivable bisphenols, are considered promising and possibly safer alternatives to bisphenol A (BPA), but comprehensive toxicity investigations are needed to ensure safety. Most toxicity studies of BPA and its analogues have focused on potential estrogenic activity, and only limited toxicological data are available on other toxicity aspects, such as genotoxicity at low exposure levels. In this study, the genotoxicity of six lignin-derivable bisguaiacols with varying regioisomer contents and degrees of methoxy substitution was investigated using a multi-tiered method, consisting of in silico simulations, in vitro Ames tests, and in vivo comet tests. The toxicity estimation software tool, an application that predicts toxicity of chemicals using quantitative structure-activity relationships, calculated that the majority of the lignin-derivable bisguaiacols were non-mutagenic. These results were supported by Ames tests using five tester strains (TA98, TA100, TA102, TA1535, and TA1537) at concentrations ranging from 0.5 pmol/plate to 5 nmol/plate. The potential genotoxicity of bisguaiacols was further evaluated using in vivo comet testing in fetal chicken livers, and in addition to the standard alkaline comet assay, the formamidopyrimidine DNA glycosylase enzyme-modified comet assay was employed to investigate oxidative DNA damage in the liver samples. The oxidative stress analyses indicated that the majority of lignin-derivable analogues showed no signs of mutagenicity (mutagenic index < 1.5) or genotoxicity, in comparison to BPA and bisphenol F, likely due to the methoxy groups on the lignin-derivable aromatics. These findings reinforce the potential of lignin-derivable bisphenols as safer alternatives to BPA.

Genotoxic hazard and oxidative stress induced by wastewater irrigated soil with special reference to pesticides and heavy metal pollution

Due to enhancement of industrial growth and urbanization, soil contamination is increasing prominently. Therefore, it is important to examine possible adverse effects of industrial waste. Soil samples were might to be polluted with several heavy-metals and pesticides. Gas chromatographic results showed occurrence of high-level of organochlorine and organophosphate pesticides in studied soil samples. Genotoxicity of soil extracts was assessed using environmental-risk assessment models. Soil samples were extracted in hexane and dichloromethane solvents and were evaluated for genotoxic potential by prokaryotic (Ames test, plasmid nicking assay and E. coli K-12 DNA repair defective mutants) and eukaryotic (Allium cepa root chromosomal aberration and Vigna radiata seed-germination test) bioassays. Strain TA98 was found the most susceptible among soil extracts. The mutagenicity of hexane soil extract from wastewater irrigation was found to be higher than that of DCM samples in terms of mutagenic index, mutagenic potential, and induction factor for Ames strains. The damage in DNA repair defective mutants of hexane extracts were found higher compared to DCM extracts at dose of 20 μl/ml of culture. Survival in polA, lexA and recA mutants were 39%, 47% and 55% while treated with hexane extract. Allium cepa test, mitotic index was decreased in dose-dependent way and various kinds of chromosomal aberrations were found. Vigna radiata seeds germination and other parameters were also affected when treated with wastewater irrigated (WWI) soil. Oxidative stress in V. radiata roots were also showed under CLS microscope. Genotoxicity of WWI soil extract was also confirmed by plasmid nicking test. Our study provides possible explanation for the assessment of potential health and environmental hazards of the industrial region.

Phytochemical Profile and Chemopreventive Properties of Cooked Glutinous Purple Rice Extracts Using Cell-Based Assays and Rat Model.

Purple rice has gained attention for its health promoting potential due to a high content of bioactive phytochemicals. The heat generated during cooking alters the quality and quantity of nutrients and phytochemicals in food. This study aimed to investigate the phytochemical profile and chemopreventive properties of cooked glutinous purple rice using cell-based assays and a rat model. Purple rice was cooked in a rice cooker and was then further extracted with solvents to obtain dichloromethane and methanol extracts. The methanol extracts of glutinous purple rice contained great amounts of phenolics, flavonoids, and anthocyanins. Protocatechuic acid (2.26–5.40 mg/g extract) and cyanidin 3-glucoside (34.3–65.7 mg/g extract) were the major phenolic acid and anthocyanin contents, respectively. After cooking, the content of anthocyanins, γ-oryzanols, and phytosterols decreased, while the amount of some phenolic acid and tocol contents increased. Methanol extracts of glutinous purple rice inhibited reactive oxygen species production about 60% in PMA-treated peripheral blood mononuclear cells, reduced nitric oxide formation in LPS-induced RAW 264.7 cells (26–39% inhibition), and exhibited antimutagenicity against several mutagens using the Ames test, but dichloromethane extracts presented only mild anti-inflammatory activities. Although methanol extracts induced mild mutagenicity (mutagenic index 2.0–2.5), they did not induce micronucleated hepatocyte formation and certain hepatic CYP450 isozyme activities in rats. However, the mutagenicity of the methanol extract significantly declined after cooking. In summary, the methanol extract of the cooked glutinous purple rice might be a promising cancer chemopreventive fraction, which was neither genotoxic nor posing adverse effects on phytochemical–drug interaction in rats.

Comparative chemical analysis, mutagenicity, and genotoxicity of Petroleum refinery wastewater and its contaminated river using prokaryotic and eukaryotic assays.

Concern on the toxicity of final wastewater generated by the petroleum refining industry has increased in recent years due to the potential health threats associated with their release into the waterways. This study determined the mutagenic and genotoxic potential of petroleum refinery wastewater and a receiving river using the Ames fluctuation test onSalmonella typhimuriumstrains TA100 and TA98, SOS chromotest onEscherichia coliPQ37, and piscine peripheral micronucleus (MN) assay. Analyses of the physicochemical parameters, heavy metal, and organic contents of the samples were also performed. Ames test result showed that the two tested samples were mutagenic with TA100 strain as the more responsive strain for both the refinery wastewater and the river sample in terms of the calculated mutagenic index. A similar result was obtained in the SOS chromotest; however, the E. coli PQ37 system recorded a slightly higher sensitivity for detecting genotoxins than the Salmonella assay in the two samples. MN data showed induction of a concentration-dependent significant (p < 0.05) increase in the frequency of MN by both samples when compared with the negative control. Generally, the refinery wastewater induced the highest mutagenicity and genotoxicity compared to the river sample in the three assays used. Haemoglobin, platelets, red blood cells, mean corpuscular volume, total white blood cells, heterophils, haematocrit, and eosinophils reduced significantly with increased lymphocytes, basophils, mean corpuscular haemoglobin, and mean corpuscular haemoglobin concentration in fishes exposed to both samples. Total petroleum hydrocarbon, benzene, toluene, phenol index, polycyclic aromatic hydrocarbons, cadmium, mercury, nickel, lead, and vanadium contents analysed in the samples were believed to be responsible for the observed genotoxicity and mutagenicity. The findings of this study revealed that petroleum refinery wastewater is a potential mutagenic and genotoxic risk to the environment.

Organic and Inorganic Pigments from Commercial Inks Used in Eyebrows’ Microblading: Approach for Assessing the Safety Use by Bacterial Reverse Mutation Assay

Background and Objective: The aesthetic products’ safety should not be neglected to the detriment of the market. This study aimed to evaluate the mutagenic potential of commercial inks. It was formulated with organic (P1) and inorganic (P2) pigments for eyebrows microblading using a validated method by regulatory agencies, the Salmonella/microsome assay, to assure the safety of use. Methods: The assay was carried out in three steps: preliminary toxicity, medium without (-S9), and presence (+S9) of metabolic activation. The strains, auxotrophic to histidine (His-) TA97a, TA98, TA100, and TA102, were exposed to both types of inks, in triplicate, compared to negative and positive controls. Data were statistically analyzed, and values with mutagenic index ≥ 2.0 were indicative of mutagenicity. Results: The inks with organic (P1) and inorganic (P2) pigments were not toxic to TA98 and TA100 S. typhimurium tester strains, even at concentrations applied in humans. Both inks were not mutagenic either in the absence or presence of metabolic activation in the tested concentrations, including that applied in humans. The assay showed that P1 and P2 were not direct (-S9) or indirect (+S9) mutagens as commercially formulated. Conclusions: These results indicate that applying these inks on organisms with microsomal enzymes, including humans, is safe since no compound in the inks became more toxic to induce the bacterial reverse mutation.

Evaluation of the Effect of Gold Mining on the Water Quality in Monterrey, Bolívar (Colombia)

Gold mining uses chemicals that are discharged into rivers without any control when there are no good mining practices, generating environmental and public health problems, especially for downstream inhabitants who use the water for consumption, as is the case in Monterrey township, where the Boque River water is consumed. In this study, we evaluate Boque River water quality analyzing some physicochemical parameters such as pH, heavy metals, Hg, and cyanide; bioassays (Lactuca sativa, Hydra attenuata, and Daphnia magna), mutagenicity (Ames test), and microbiological assays. The results show that some physicochemical parameters exceed permitted concentrations (Hg, Cd, and cyanide). D. magna showed sensitivity and L. sativa showed inhibition and excessive growth in the analyzed water. Mutagenic values were obtained for all of the sample stations. The presence of bacteria and somatic coliphages in the water show a health risk to inhabitants. In conclusion, the presence of Cd, Hg, and cyanide in the waters for domestic consumption was evidenced in concentrations that can affect the environment and the health of the Monterrey inhabitants. The mutagenic index indicates the possibility of mutations in the population that consumes this type of water. Bioassays stand out as an alert system when concentrations of chemical contaminants cannot be analytically detected.

Genotoxicity and mutagenicity of particulate matter emitted from diesel, gas to liquid, biodiesel, and farnesane fuels: A toxicological risk assessment

The genotoxic and mutagenic potential of soluble organic material extracted from particulate matter emitted by a typical automotive diesel engine was evaluated from the combustion of different alternative and/or renewable fuels. One of these fuels, called Farnesane, is a novel renewable paraffinic hydrocarbon obtained from sugar cane using a biotechnology fermentation process. Tests were carried out under a steady state condition with high level of generated particulate matter, which was collected with a stainless-steel filter located 1.5 m downstream of the exhaust manifold. The mutagenicity index was evaluated using an Ames test, obtaining values for Farnesane and biodiesel fuels lower than those for fossil fuels. However, all fuels showed a mutagenicity index higher than the Phosphate Buffered Saline control. The soluble organic material was extracted from particulate matter to evaluate its genotoxicity activity using an alkaline comet assay in human lymphocytes. The obtained genotoxicity trends were similar to mutagenicity, biodiesel and Farnesane fuels showing the lowest toxicity impact. Additionally, the cell viability was studied and classified according to the type of damage. The lowest values of average Deoxyribonucleic Acid migration and cell damage associated to Farnesane, together with its larger particle size, makes this renewable fuel a potential substitute of the traditional diesel.

Comparison of Impact of Residual Radioactivity Versus Pesticide-Derived Soil Pollution on Genotoxicity

A comparative analysis was performed of the impact on living organisms, in terms of genotoxicity, of two factors, namely, radiation (residual 137Cs) and a chemical factor (pesticides) . The experiment was conducted both on individual pesticide samples and the aqueous extracts of soil samples. Soil samples were collected in the territory of the Orel region (Russia) from May 12 to May 15, 2010, i.e., years after the Chernobyl accident. Changes in the mutagenic index derived from Ames assay (Salmonella/microsomes) and the data obtained from mammalian cell culture in the Cricetulus griseus test (Chinese hamster) of the soil samples arising from the cesium radioactivity and the content of pesticides in these samples were considered. No direct correlation was found between dose level and genotoxicity at low radiation doses (20 μR/h), i.e., correlation with DNA mutations produced, both in terms of frameshifting and the base replacement type. On the contrary, a clear tendency was observed for increase in genotoxicity with increase in the concentration of pesticides and the entire amount of anthropogenic toxicants in the studied soil samples.

Anti-HIV-1 activity and safety profile of a polyherbal gel formulation as a candidate microbicide

Microbicides represent one of the promising options for the prevention of sexually transmitted HIV infection. Medicinal plants have a long history of use to provide treatment against many diseases. In this regard, polyherbal aqueous gel formulation comprising 50% ethanolic extract prepared from the heart wood of Acacia catechu, leaves of Lagerstroemia speciosa, fruits of Aegle marmelos, Phyllanthus emblica and Terminalia chebula using carbopol 974 P NF polymer has been prepared. The gel formulation showed significant anti-HIV-1 activity against both CXCR4 tropic (HIV-1NL4.3) and CCR5 tropic (HIV-1NLAD8) viruses with IC50 values of 58.17 ± 4.4 and 63.54 ± 6.8 μg/ml respectively. It also showed significant inhibition of HIV-1 reverse transcriptase (p = 0.004), protease (p = 0.014), and integrase (p = 0.018) enzymes activity as compared to gel base. The gel formulation did not show any adverse effect on the viability of the vaginal lactobacilli, human vaginal keratinocytes (Vk2/E6E7), and the monolayer formed by Caco-2 cells. Furthermore, no significant changes in the secretion of pro-inflammatory cytokines such as IL-1β, IL-6, IL-8, IL-10, IL-12p70, and TNF from the gel formulation treated Vk2/E6E7 cells were observed as compared to cells treated with gal base or medium control. Gel formulation did not show any significant increase in mutagenic index using Salmonella typhimurium. Gel formulation stored at either 30 ± 2 °C or 40 ± 2 °C was stable for 6 months for anti-HIV-1 activity. In conclusion, the polyherbal gel formulation reported herein having potent anti-HIV-1 activity and with limited safety profile may be a promising candidate as an herbal microbicide.

In vitro mutagenicity and genotoxicity of raw and simulated leachates from plastic waste dumpsite

Increase in production of different types of plastics has led to increase in the amount of plastic waste generation worldwide. The chemical constituents of these plastic wastes have made their disposal an important economic and environmental health problem globally. This study assessed the mutagenic and genotoxic potential of plastic waste dumpsite raw and simulated leachates using the Ames Salmonella fluctuation test with Salmonella typhimurium strains TA98 and TA100, and the SOS chromotest with Escherichia coli PQ37. Physico-chemical parameters and organic constituents of the leachates were also analyzed. The result of the Ames test showed that the leachates are mutagenic even at low concentration. Also, the TA100 strain was the more responsive strain in terms of mutagenic index in the absence of metabolic activation. The SOS chromotest results complimented the Ames Salmonella fluctuation test results. Nevertheless, the E. coli PQ37 system was slightly more sensitive than the Salmonella assay for detecting mutagens and genotoxins in the tested leachates. Generally, simulated leachate showed a higher mutagenicity and genotoxicity than the raw leachate. Pb, Cd, Cr, Ni, Cu, As, PBDEs, PAHs, PCBs, and Bisphenol A contents analyzed in the leachates were believed to play significant role in the observed mutagenicity and genotoxicity in the microbial assays. These data showed that plastic waste constituents are capable of inducing DNA damage in exposed organisms and might induce similar damage in plants, animals and humans exposed to it, hence, great care should be taken to eliminate indiscriminate disposal of plastics in the environment.

Dietary Heterocyclic Amine Intake and Colorectal Adenoma Risk: A Systematic Review and Meta-analysis.

Heterocyclic amines (HCA) are potent carcinogenic substances formed in meat. Because of their mutagenic activity, they may increase the risk of colorectal adenomas, which are the precursors of colorectal cancer, one of the most prevalent cancers worldwide. The aim of this meta-analysis was to synthesize the knowledge about the intake of HCAs and its associations with CRA. We conducted a systematic search in PubMed and EMBASE. We used odds ratios (OR); or relative risks, RR) from every reported intake and compared the highest versus lowest level of dietary HCAs. In addition, we assessed a dose-response relationship. Twelve studies on HCA intake and risk of CRA were included in our analysis. We observed a statistically significant association when comparing top versus bottom intake category of 2-amino-1-methyl-6-phenylimidazo[4,5-b]pyridine [PhIP; OR = 1.20; 95% confidence interval (CI) = 1.12-1.29], 2-amino-3,8-dimethylimidazo[4,5-f]quinoxaline (MeIQx; OR = 1.20; 95% CI = 1.08-1.34), 2-amino-3,4,8-trimethylimidazo[4,5-f]quinoxaline (DiMeIQx; OR = 1.16; 95% CI = 1.05-1.27), benzo(a)pyrene (BaP; OR = 1.15; 95% CI = 1.04-1.27), and mutagenicity index (OR = 1.22; 95% CI = 1.06-1.41). Furthermore, we observed a significant dose-response effect for PhIP, MeIQx, and mutagenicity index. This meta-analysis suggests that there is a positive association of HCAs, BaP, mutagenicity index with risk of CRA. In addition, our dose-response analyses showed an increased risk of CRA for PhIP, MeIQx, and mutagenicity index. This study provides evidence for a positive association between the dietary intake of meat mutagens and CRA risk.

Significant increase in the dominant lethal mutations: benzo(a)pyrene

Background: Use of biomarkers particularly under in vivo condition is of importance in risk assessment. This study is proposed to identify the in vivo antigenotoxic potential using dominant lethal mutation test in B(a)P exposed, turmeric fed animals and evaluate the role of turmeric in counteracting the germ cell mutations. Methods: Twenty four male mice were taken and divided into 4 groups each group containing 6 mice/group. The first and second group received stock diet (control) while the third and fourth groups received 5% turmeric diet for a period of one month. At the end of feeding period, second and fourth group received a single dose of benzo(a)pyrene 1mg/mouse intraperitoneally. After 1 week each male mouse was mated to 3 female mice at 1,4, 8 and 12 week intervals. On 13th day from mid point of mating, the females were sacrificed and live and dead embryos were counted to study the pre and post implantation loss of embryos. The differences in the frequencies of pre and post implant deaths were done by comparing treated with control group at 4-time periods by analysis of variance (ANOVA) and chi-square test which is used to compare treatment group and positive control with negative control. Results: Feeding of 5% turmeric and treatment with a single dose of B(a)P did not show any significant effect on the mutagenic index or the frequency of pregnancy in treated groups compared with control. There was no apparent induction of dominant lethal mutations in B(a)P or B(a)P+Turmeric treated groups. No post implantation dominant lethal effects produced by B(a)P could be detected in this study. The induction of B(a)P and treatment with turmeric did not influence the dominant lethal test. Conclusion: B(a)P did not produce a significant increase in the dominant lethal mutations. Turmeric also did not give any positive response and may be considered as non-mutagenic. The negative result suggests that under the conditions of the test the test substance may not be genotoxic in the germ cell of the treated sex of the test species.

Herbal Gel Formulation Developed for Anti-Human Immunodeficiency Virus (HIV)-1 Activity Also Inhibits In Vitro HSV-2 Infection.

Herpes simplex virus-2 (HSV-2) infection is the most common cause of genital ulcers. The impact of ulcers also demonstrates a strong link to the human immunodeficiency virus (HIV) infection. Complications, drug resistance, and side-effects of anti-viral drugs make the treatment of HSV-2 infection challenging. Herbal medicines have shown potential against HSV-2 and HIV infections. In this context, polyherbal gel formulation comprising 50% ethanolic extracts from Acacia catechu, Lagerstroemia speciosa, Terminalia chebula and Phyllanthus emblica has been developed. The gel formulation significantly exhibited virucidal activity against both HIV-1 and HSV-2 infections with IC50, 55.93 ± 5.30 µg/mL and 27.26 ± 4.87 µg/mL, respectively. It also inhibited HSV-2 attachment and penetration to the Vero cells with an IC50 = 46.55 ± 1.25 µg/mL and 54.94 ± 2.52 µg/mL respectively, which were significantly lower than acyclovir. However, acyclovir is more potent in post-infection assay with an IC50 = 0.065 ± 0.01 µg/mL whereas gel formulation showed an IC50 = 469.05 ± 16.65 µg/mL under similar conditions. Gel formulation showed no inhibitory effect on the viability of lactobacilli, human vaginal keratinocyte cells (Vk2/E6E7), and the integrity of the Caco-2 cells monolayer. Gel formulation did not lead to any significant increase in the secretion of pro-inflammatory cytokines and mutagenic index. The proposed gel formulation may be a promising candidate microbicide for the prevention of sexually transmitted HIV-1 and HSV-2.

Modified tannins and their application in wastewater treatment.

The bio-flocculants used in this study were synthesised by the Mannich reaction, which includes three reagents: a substrate (tannin extracts of Acacia, Quebracho, and Castanea), formaldehyde, and an amine derivative (ethanolamine, diethanolamine, ammonium chloride). Nine natural flocculants were prepared by combining extracts and amines; these products were evaluated in three different wastewater samples in two experimental phases. In phase I, five physicochemical parameters were analysed. From the data obtained, a multivariate, completely randomised design (CRD-Manava) was used, with a factorial arrangement and mean plots. In phase II, the three bio-flocculants with the most statistically significant responses and their mixtures were examined, evaluating 14 biological and physicochemical parameters. Statistical analysis was guided in this phase by CRD blocks, finding a significant removal in the physicochemical parameters analysed in the different types of wastewater and obtaining removal rates between 50 and 90%, depending on the parameter. At the end of both phases, the bio-flocculants acacia-ammonium chloride and quebracho-diethanolamine were the most efficient in the removal of turbidity (34-99%), true colour (93-100%) and total solids (12-99%). In addition, the natural flocculants showed low mutagenicity index (MI: 0.33-0.93) compared to aluminium sulphate (MI: 4.87-8.81).

Genetic damage induced by electronic waste leachates and contaminated underground water in two prokaryotic systems

The inappropriate and unsafe management practices related to disposal and recycling of electronic wastes in Nigeria has led to environmental and underground water contamination. Reports on the level and type of contamination as well as the possible DNA damage effects of this contamination are insufficient. This study evaluated the DNA damaging potential of e-waste simulated and raw leachates, and its contaminated underground water using the SOS chromotest on Escherichia coli PQ37 and the Ames Salmonella fluctuation test on Salmonella typhimurium strains TA98 and TA100, without and with metabolic activation. Physico-chemical parameters of the samples were also analyzed. The result of the Ames test showed induction of base pair substitution and frameshift mutation by the test samples. However, the TA100 was the more responsive strain for the three samples in terms of mutagenic index in the absence and presence of metabolic activation. The SOS chromotest results were in agreement with those of the Ames Salmonella fluctuation test. Nevertheless, the E. coli PQ37 system was slightly more sensitive than the Salmonella assay for detecting genotoxins in the tested samples. Lead, cadmium, manganese, copper, nickel, chromium, arsenic, and zinc contents analyzed in the samples were believed to play a significant role in the observed DNA damage in the microbial assays. The results of this study showed that e-waste simulated and raw leachates, and its contaminated underground water are of potential mutagenic and genotoxic risks to the exposed human populace.

Genotoxic and cytotoxic potential of whole plant extracts of Kalanchoe laciniata by Ames and MTT assay.

Lack of data on safety of herbal medicines have endangered human health and life. The present study evaluated the genotoxic and mutagenic effect of Kalanchoe laciniata to access the safety and usefulness of the medicinal plant. Aqua-methanolic and n-hexane extracts of K. laciniata were evaluated for the genotoxic potential using Ames assay and cytotoxicity was evaluated using MTT assay. Ames assay was conducted using two strains of Salmonella typhimurium TA-100 and TA-102 whereas MTT assay was performed on baby hamster kidney cell line BHK-21. Aqua-methanolic extract of K. laciniata exhibited significant mutagenicity when exposed to TA-102 strain with a mutagenic index of 50.66 and 54.74 at maximum dose 150 mg/plate. The extract was also mutagenic to TA-100 strain but to a lesser extent. M.I of n-hexane extract was 12.15 and 15.51 for TA-100 and TA-102 respectively. n-hexane extract was mutagenic but little difference was observed between results of two strains. Both extracts were found to be cytotoxic with an IC50 of 321.9 and 638.5 µg/mL for aqua-methanolic and n-hexane extracts respectively. On the basis of results it was concluded that aqua-methanolic and n-hexane extracts of K. laciniata possess mutagenic and cytotoxic potential. It is suggested to explore the plant further to evaluate its safety in rodents and other species.

Mutagenicity of Wastewater Extracts from Pulp and Paper Industry

Wastewater samples were collected from pulp and paper mill located in Kashipur (India) and were extracted using dichloromethane (DCM), chloroform and hexane solvents (all were HPLC-grade). The extracts were assayed for their mutagenic potential using Ames Salmonella mutagenicity assay. TA98 strain was found to be the most responsive, in terms of induction factor (Mi) and slope (m) of the initial linear dose-response curve as determined by linear regression analysis up to the increasing doses indicating the presence of frame shift mutagens in the test samples. Mutagenicity of different extracts is arranged as follows: dichloromethane extracted water samples > hexane extracted water samples > chloroform extracted water samples. Hexane extract exhibited maximum mutagenic index of 13.0 and induction factor (Mi) 2.48 with TA98. The order of responsiveness based on the mutagenic index and induction factor for the test samples was in the following order: TA98 > TA97a > TA100 > TA102 > TA104. Our findings suggest that TA97a, TA98, TA100, TA102, TA104 were sensitive towards the wastewater extracts and showed considerable mutagenicity.