Micronucleus Assay with Tetrad Cells of Tradescantia.

The Tradescantia micronucleus assay has been used since 50 years for the detection of genotoxins (including carcinogens) in the environment. A large database concerning the effects of individual chemicals and complex environmental mixtures (soil, air and waters) has accumulated. In contrast to other mutagenicity test systems, the effects of low concentrations of heavy metals, radionuclides, certain herbicides, pesticides and gaseous mutagens can be detected and it is also possible to conduct in situ biomonitoring studies with plant. The test system has been validated and standardized protocols have been developed for laboratory experiments and for field studies which are described in this chapter.

Assessment of the genotoxic potential of water courses impacted by wastewater treatment effluents using micronucleus assay in plants from the species Tradescantia.

Water pollution and the increase in genotoxic consequences in aquatic environments are well documented indicating the necessity and importance of biomonitoring programs. The objective of the present study was to determine the environmental quality of water resources and genotoxic potential of materials present within water samples obtained from the Perdizes River and the Mumbuca Stream, located in a region of discharge of wastewater treatment effluents using Tradescantia micronucleus assay (Trad - MCN). Water samples were collected from different locations up and downstream of the wastewater treatment plant during rainy season and subsequently submitted to physico-chemical analysis and Trad-MCN bioassay. The spatial distribution of the physico-chemical parameters assessed suggested that discharges of wastewater treatment effluents reduced water quality at all sites examined. Further, exposure to wastewater treatment effluents produced genotoxic effects on tetrads of Tradescantia pallida. These results reinforce the sensitivity of the Trad-MCN bioassay and its potential application in water quality monitoring programs concomitant with physicochemical evaluation.

Genotoxicity and carcinogenicity of ivermectin and amoxicillin in vivo systems.

Antiparasitic substances are chemicals used to control or kill endoparasites and ectoparasites. Based on the premise that Ivermectin (IVM) and Amoxicillin (AMX) are commonly considered in parasitic control in mammals, the present study aimed to evaluate the carcinogenic and genotoxic potential of different concentrations of IVM and AMX through the detection of epithelial tumor test in Drosophila melanogaster. Third-instar larvae descending from the cross between wts/TM3, Sb1 females and mwh/mwh males were treated with different concentrations of IVM (2.9, 5.8, 11.6 and 23.2 x 10-17 mM) or AMX (1.37, 2.74, 5.48 and 10.9 x 10-16mM). The results revealed that IVM increased the frequency of epithelial tumor in D. melanogaster considering all evaluated concentrations, while AMX showed no carcinogenic effect. Furthermore, the Micronucleus (MN) test in Tradescantia pallida was used to evaluate the genotoxic effect of IVM and AMX. T. pallida individuals were exposed for 8 hours at different concentrations of IVM (5.71, 11.42, 22.84 and 45.68 x 10-5mM) or AMX (5.13, 10.26, 20.52 and 41.05 x 10-3mM). Findings showed an increase in the frequency of micronuclei in T. pallida treated with 11.42, 22.84 and 45.68 x 10-5mM of IVM. We conclude that chronic exposure to IVM is directly associated with events resulting from genetic instability (genotoxicity and carcinogenicity). On the other hand, AMX was neither carcinogenic nor genotoxic for D. melanogaster and T. pallida.

Evaluation of the genotoxicity of neurotoxic insecticides using the micronucleus test in Tradescantia pallida.

Neonicotinoids and phenylpyrazoles are classes of neurotoxic insecticides which are able to bind at different ligand sites of neural receptors, leading to the deregulation of insect neural activity and hence resulting in death. The misuse or indiscriminate use of these chemicals is directly associated with several toxicological effects in biota and at different trophic levels. Based on this premise, the aim of the present study was to evaluate and compare the genotoxic capacity of different concentrations of thiamethoxam (TMX), acetamiprid (ACP), imidacloprid (IMI) and fipronil (FP) through the Micronucleus Test in Tradescantia pallida (Trad-MCN). After acclimatization (24 h), T. pallida stems were treated with stablished concentrations of TMX, ACP, IMI and FP for 8 h. Then, the stems of the model organism were submitted to a recovery phase (24 h). The young inflorescences were harvested and fixed in Carnoy solution and, after 24 h, were conserved in ethanol 70% until the analyzes. The obtained anthers were macerated on slides for microscopy, stained with acetic carmine dye and covered with coverslips before analysis by light microscopy. Considering the insecticides, the micronuclei (MN) frequency in plants treated at concentrations of 0.2 and 0.4 g L-1 for TMX, 0.2; 0.4 and 0.8 g L-1 for ACP, 0.1; 0.2; 0.4; 0.8 and 1.6 g L-1 for IMI and 0.2; 0.4; 0.8 and 1.6 g L-1 for FP differed statistically (p < 0.05, Tukey) from the MN frequency of the negative control. All chemicals evaluated revealed genotoxic activity in T. pallida at the highest concentrations.

Genotoxic and mutagenic assessment of spinosad using bioassays with Tradescantia pallida and Drosophila melanogaster.

Spinosad (SPN) is a naturally-occurring insecticide obtained from the fermentation process of the actinomycete Saccharopolyspora spinosa. Owing to the larvicidal action, the compound has been used in the control of Aedes aegypti. As a new insecticide commercially available in the market, few data are reported on genotoxic effects in non-target organisms. The objective of the present study was to evaluate the mutagenic effect of SPN through the Micronucleus Test in Tradescantia pallida (Trad-MCN) and using the mutation and somatic recombination test in Drosophila melanogaster (SMART). At the Trad-MCN, after acclimatization (24 h), T. pallida stems were submitted to chronic treatment with SPN at concentrations of 0.156; 0.312; 0.625; 1.25 and 2.5 g/L solution for 24 h, followed by a recovery period. In SMART, considering the third stage larvae, offspring resulting from the ST and HB crossing were placed on chronic treatment (48 h) with 0.039; 0.078 and 0.156 µg/mL of SPN solution. No mutagenic effect was observed at any of the evaluated concentrations in SMART. Additionally, SPN is more toxic after metabolism via CYP6A2 (cytochrome P450) in D. melanogaster. However, SPN at the concentrations of 0.625; 1.25 and 2.5 g/L was able to induce high frequency of micronuclei in T. pallida. Under the experimental conditions of T. pallida in the present study, SPN caused genotoxic activity.

Response of Tradescantia plants to oxidative stress induced by heavy metal pollution of soils from industrial areas.

Numerous investigations have demonstrated that even soil in which concentrations of individual elements do not exceed permissible limits can cause harmful effects in living organisms. In the present study, polluted-soil-induced oxidative stress was evaluated using Tradescantia clone 4430, which is widely used for genotoxicity evaluations, employing biochemical (superoxide dismutase (SOD), contents of ascorbic acid (AA), carotenoids (Car), hydrogen peroxide (H2O2), chlorophyll (Chl) a/b ratio), and molecular (RAPD and differential display (DD-PCR)) markers after long-term exposure. The activity (staining intensity) of SOD isoforms in Tradescantia leaves was higher in plants grown in all heavy-metal-polluted test soils compared to the control. No direct link between the soil pollution category and the contents of AA, Car, Chl a/b in Tradescantia leaves was revealed, but the concentration of H2O2 was shown to be a sensitive biochemical indicator that may appropriately reflect the soil contamination level. Both short-term (treatment of cuttings with H2O extracts of soil) and long-term (0.5 and 1.0 year) exposure increased MN frequencies, but the coincidence of the MN induction and the soil pollution level was observed only in some cases of long-term exposure. Soil (geno)toxin-induced polymorphism in the RAPD profile was determined with two primers in plants after long-term exposure to soils of an extremely hazard category. Transcript profiling of plants after long-term cultivation in test soils using DD-PCR showed that the majority of differentially expressed transcript-derived fragments (TDFs) were homologous to genes directly or indirectly participating in photosynthesis, the abiotic stress response, and signal transduction cascades.

Tradescantia as a biomonitor for genotoxicity evaluation of diesel and biodiesel exhaust emissions.

Biodiesel, an alternative energy source, is promoted as cleaner and safer than other fuel options due to its reported reduction of particulate and gaseous emissions (CO2, CO, and total hydrocarbons). However, its volatile organic compounds (VOCs) and polycyclic aromatic hydrocarbon (PAHs) emissions are key to understanding its toxic, mutagenic and carcinogenic risk factors. This research was developed to assess the genotoxic impact of exhaust emissions using biodiesel from animal fat, palm oil and soybean oil blended with diesel (B80). Diluted exhaust gases were analyzed simultaneously for pollutant emissions and for toxicity using an exposure chamber called the BioToxMonitor, where Tradescantia pallida and a KU-20 clone were exposed to exhaust following Trad-MCN and Trad-SH bioassays. The results show differences in the emission compositions and considerable mutagenic potential among the three biodiesels tested, with palm oil biodiesel emissions being the least harmful, based on its low pollutant concentrations and the negative response in the TradSH bioassay. In contrast, the animal fat biodiesel and soybean oil biodiesel emissions were as toxic as the diesel emissions, being positive in both Trad bioassays. This could be related to the PAH and carbonyl concentrations found in the vehicular exhaust. The genotoxicity of diesel emissions was related to PM1 and the concentrations of both gas and particle PAHs concentrations, which were two times higher compared to the highest concentrations observed for biodiesel. The data suggest that micronucleus assays in Tradescantia pallida are more sensitive for gaseous pollutant exposure. This is the first reported study of biodiesel exhaust biomonitoring in situ and under controlled conditions inside an exposure chamber.

Clastogenecity evaluation of water of Lake Sevan (Armenia) using Tradescantia micronucleus assay.

The clastogenic effects of water samples in seven locations of Lake Sevan (Armenia) with the application of Trad-MCN (micronuclei) bioassay using Tradescantia (clone 02) were investigated. A significant increase in the frequency of micronuclei in tetrads of pollen microspores and tetrads with micronuclei exposed to the test samples compared to the control has been revealed. A multivariate analysis indicated linkage between the frequencies of occurrence of micronuclei in the cells and Ni and Co ions. The results were compared with the endpoints of another Tradescantia-based test system (stamen hair mutation test) performed on the same water samples and generation of the plant: occurrences of micronuclei in sporogenic cells coincided with that of non-surviving stamen hair.

Evaluation of vehicular pollution using the TRAD-MCN mutagenic bioassay with Tradescantia pallida (Commelinaceae).

Biomonitoring is one of the tools used to assess the mutagenic potential of the atmosphere. In this study, the mutagenicity of Tradescantia pallida, a species of plant largely present in urban environments, was investigated. The objectives of this study was to estimate the mutagenic potential of vehicular flow through the TRAD-MCN bioassay in cities located at different altitudes in the southwest mesoregion of Mato Grosso do Sul, Brazil, to infer possible abiotic agents that may contribute to the effects of atmospheric pollutants, and finally to map the cities with greater risks to the health of the local population. To achieve these objectives, the Tradescantia-micronucleus test was performed on young buds of T. pallida collected between August 2015 and August 2016 in nine cities of Mato Grosso do Sul. These buds were exposed to traffic flows of various intensities. The data collected consisted of measurements of meteorological parameters and vehicular traffic counts for each city. The variables considered were: mean ambient temperature; micronuclei frequency; vehicular flow; altitude; relative humidity; pluviosity. The application of the Trad-MCN bioassay, with the consideration of environmental variables and altitudes, and the use of the Kernel interpolation technique, allowed us to map the areas with significant pollution risks to the population. The highest frequency of exposure to mutagens occurred in the cities with the highest vehicular traffic intensity. The average ambient temperature failed to show a linear association with the frequency of the micronuclei in the samples analyzed (r = 0.11ns). A positive correlation was observed between micronuclei frequency and vehicular flow, (r = 0.67; p ≤ 0.001%) and between micronuclei frequency and altitude (r = 0.24; p ≤ 0.05). A negative correlation was found between relative humidity and micronuclei frequency (r = -0.19; p ≤ 0.05%). Thus, higher micronuclei frequency tended to be present in locations with low relative humidity and high altitudes and vehicular flow.

Metal bioaccumulation and mutagenesis in a Tradescantia clone following long-term exposure to soils from urban industrial areas and closed landfills.

Soil mutagens, particularly metals, may persist long after the source of pollution has been removed, representing a hazard to plants, animals, and humans in or near contaminated areas. Often, due to urban growth, previous land uses may be forgotten and hazards overlooked. We exposed Tradescantia clone #4430 plants to soil from two industrial areas (with different former uses) and two urban waste landfills in the city of Vilnius, all of which were long disused. Two modes of exposure were used: long-term exposure of growing plants in test soils for 0.5 or 1.0y, and short-term exposure of cuttings to water and dimethyl sulfoxide (DMSO) soil extracts. An increased frequency of micronuclei (MN) was observed with both modes of exposure. The concentrations of 24 metals and other elements were analyzed in the test soils and in above-ground plant parts, under both exposure modes, and the concentration coefficients (Cc) for various elements, the total contamination index (Zs) for soils and plants, and the bioaccumulation factor (BAF) for plants were calculated. These measurements allow a comparison of the contamination levels of soils and plants with equalized values. Metal accumulation levels in plants and soils showed significant differences, providing a better understanding of the genotoxicity of soils from closed landfills and highlighting the need to determine the concentrations of metals and other genotoxicants in plants in relation to genotoxicity.

The ecogenotoxic plant biomonitoring of a long-term polluted area in central Slovakia.

The aim of the study was to analyze, update, and complete the results of research in the field of in situ phytoindication of environmental genotoxicity near the aluminum plant in Ziar nad Hronom in central Slovakia. The authors focused on two methodologies: pollen abortivity assay of native flora and Trad-MCN assay. Comparison of changes in responses of living systems to changes in precipitation was conducted, and it suggests that there is an existence of an impact from a dilution effect to the plants. Also, the gradual increase of pollen abortivity in the 1990s and its decrease from 2009 were observed in a majority of species of wild flora. On an annual basis, abortivity has declined, although it has risen up slightly within each season. Despite a gradual decrease in the micronucleus frequency, the study area is still influenced by the ecogenotoxic factors. Pollen analysis of native flora and introduced Tradescantia plants indicates the long-time presence of ecogenotoxicity in this region due to the presence of aluminum plant.

Assessment of mutagenicity of water from Lake Sevan, Armenia with application of Tradescantia (clone 02).

For many decades water resources in Armenia have been affected by anthropogenic activity, therefore, a regular bioindication of genotoxic effects of the water bodies is desirable. The genotoxicity of water samples collected from different parts of Lake Sevan were assessed by means of Trad-SHM (stamen hair mutation) assay using Tradescantia (clone 02). Here we report a significant increase in the frequency of somatic mutations and morphological changes in the Tradescantia inflorescences exposed to the water samples compared to the control. The somatic mutations (recessive mutation and white mutation events) were mostly linked to the concentration of Al, Ni, As, Co and Pb in Artanish, Tsapatakh and Karchaghbyur, Noradus, Martuni and Litchk, while morphological changes (non-surviving hairs) were related to Co level in Tsapatakh and Karchaghbyur. The results obtained show that Lake Sevan contains substances which may cause genotoxicity and teratogenicity in Tradescantia and probably also in aquatic animals. The results also show that Trad-SHM assay can be used for monitoring natural resources.

Evaluation the urban atmospheric conditions in different cities using comet and micronuclei assay in Tradescantia pallida.

In the present study, genotoxicity and mutagenicity were investigated in Tradescantia pallida exposed to vehicular traffic at different sites in a high-altitude tropical climate. During March, May, July, September, and November 2014, a comet assay and micronucleus bioassays were conducted on young inflorescences and leaves of T. pallida collected from twelve towns in the southern region of Mato Grosso do Sul with different amounts of vehicular traffic. Weather parameters (temperature, relative humidity and rainfall) were measured and vehicles were counted to determine traffic levels in each town. A higher frequency of genotoxic and mutagenic damage was observed in the municipality of Dourados. The highest frequency of genetic damage was observed in September and November according to both assays. Relative humidity and rainfall were inversely proportional to the frequency of genetic damage in T. pallida during the collection period. Based on these results, we conclude that the bioassays are efficient for assessing the effects of vehicular traffic in these towns with respect to weather conditions over time. These bioassays can be applied to identify risk areas, which are determined by climatic conditions and air pollutants released.

Biomonitoring of genotoxic effects and elemental accumulation derived from air pollution in community urban gardens.

Urban gardening is a growing global phenomenon with a positive impact on society. Despite several associated benefits, growing vegetables in urban gardens that are localized in highly polluted areas poses questions about the safety of the produced food. Therefore, the identification of risk factors that result in possible deleterious effects to human health is important for realizing all of the benefits to society. We evaluated the use of two biomonitoring methods in ten urban gardens of Sao Paulo city and one control site: the micronuclei frequencies for early tetrads of Tradescantia pallida (Rose) Hunt. cv. "Purpurea" Boom (hereafter, Trad-MCN) as a short-term indicator of genotoxic response and tree barks to quantify the accumulation of traffic-related chemical elements as a long-term biomarker of air pollution in urban gardens. Mature plants of Tradescantia pallida were exposed in each garden, and their inflorescences were sampled over three months. A random set of 300 early tetrads in 13 to 21 slides per garden were evaluated for micronuclei frequencies. Elemental concentrations in 428 tree barks samples from 107 different trees in the areas surrounding urban gardens were quantified using an energy dispersive X-ray fluorescence spectrometer. The frequency of Trad-MCN has a significant correlation with traffic variables and chemical elements related to road dust and tailpipe emissions deposited in tree barks. Negative associations between Trad-MCN and both the distance through traffic and the presence of vertical obstacles were observed in the community gardens. The Mn/Zn concentrations in tree barks were associated with increased Trad-MCN.

Tradescantia as a biomonitor for pesticide genotoxicity evaluation of iprodione, carbaryl, dimethoate and 4,4'-DDE.

There is a current tendency to develop and apply environmentally friendly techniques that meet the requirements of green analytical chemistry as an alternative to conventional analytical methods. For toxicity evaluation, these alternatives may be found in bioassays such as Tradescantia. This technique, developed in the 1980s, is highly sensitive to evaluate environmental mutagens, simple and cheap. In this paper, the sensibility of both the Tradescantia micronucleus bioassay (Trad-MCN) and the Tradescantia stamen hair bioassay (Trad-SH) were studied for carbaryl, dimethoate and iprodione, common agricultural and domestic pesticides that are currently used in Chile, which have never been tested with such bioassays. Biomonitor exposures were performed by capillary absorption for each individual pesticide over a wide range of concentrations, from maximum residue limits (trace levels) up to the application dose in agricultural fields. In addition, the organochloride 4,4'-DDE was included but only in the concentration range from 0.01mgL-1 to 1mgL-1, mimicking residue concentrations since it is not a commercial product but, rather, the main breakdown product of the persistent organochloride pesticide 4,4-DDT, whose use was discontinued in Chile in the 1980s. The Trad-MCN bioassay revealed a significant increase in micronucleus frequency at the early tetrads of meiotic pollen mother cells of the biomonitor Tradescantia pallida var. purpurea, induced by 4,4'-DDE (for 1mgL-1), dimethoate (for 40mgL-1, 200mgL-1, 400mg/L-1) and carbaryl (for 889mgL-1). Iprodione did not generate any significant change at the tested concentration. Meanwhile, the Trad-SH bioassay was carried out by analysis of the phenotype variations of the stamen hair cells of the Tradescantia clone KU-20 for the same pesticides and doses. This bioassay was not sufficiently sensitive for toxicity evaluation of most of the pesticides tested, with exception of dimethoate in low doses (2 and 5mg/L-1).

Genotoxicity evaluation of two metallic-insecticides using Allium cepa and Tradescantia pallida: A new alternative against leaf-cutting ants.

In order to combat leaf-cutting ants, the pesticide sulfluramid used to be the most widely utilized active ingredient. However, its use was banned in 2009 by the Stockholm Convention, although some countries were allowed to continue using it. As an effective alternative to its replacement, researchers developed a metallic-insecticide system, which is a natural product linked to metal complexes. Thus, the aim of this study was to evaluate the ability of these new metallic-insecticides in change the genetic material of non-target organisms. The tests were performed utilizing chromosomal aberrations and micronucleus tests in the Allium cepa test system and the Trad-MCN test in Tradescantia pallida. To better understand the results, one of the components of the formula, 5-methyl-phenanthroline, was also analyzed according to the same parameters. To A. cepa, the results showed that one of the metallic insecticides induced cytotoxicity and genotoxicity at different concentrations, while the other metallic-insecticide showed chromosomal instability only at the highest concentration. The analysis of 5-methyl-phenanthroline revealed that it can be related with the positive results, since genotoxic effects were induced. In the Trad-MCN test, none of the metallic-insecticides showed genotoxic activity, although one of them induced more micronucleus formation.

Chemical and toxicological characterisation of anticancer drugs in hospital and municipal wastewaters from Slovenia and Spain.

Anticancer drugs are continuously released into hospital and urban wastewaters, where they, most commonly, undergo conventional treatment in wastewater treatment plants (WWTPs). Wastewaters contain complex mixtures of substances including parent compounds, their metabolites and transformation products (TPs). In this study, samples of hospital effluents and WWTP influents and effluents from Slovenia and Spain were analyzed for twenty-two selected anticancer drugs, their metabolites and transformation products. Acute and chronic toxicity tests were performed on the crustacean Ceriodaphnia dubia, genotoxicity was determined with Tradescantia and Allium cepa micronucleus (MN) assays and in vitro comet assay in zebrafish (Danio rerio) liver cell line (ZFL cells). Sixty of the two hundred-twenty determinations revealed detectable levels of anticancer drug residues. Among the targeted compounds, platinum based were most frequently detected (90%). Furthermore, erlotinib was detected in 80%, cyclophosphamide and tamoxifen in 70% and methotrexate in 60% of the samples. Seven of ten samples were toxic to C. dubia after acute exposure, whereas after chronic exposure all samples reduced reproduction of C. dubia at high sample dilutions. Allium cepa proved insensitive to the potential genotoxicity of the tested samples, while in Tradescantia increased MN frequencies were induced by a hospital effluent and WWTP influents. In ZFL comet assay all but one sample induced a significant increase of DNA strand breaks. Correlations of chemotherapeutics or their TPs were detected for all bioassays except for Allium cepa genotoxicity test, however for each test the highest correlations were found for different substances indicating differential sensitivities of the test organisms.

Biomonitoring of the environmental genotoxic potential of emissions from a complex of ceramic industries in Monte Carmelo, Minas Gerais, Brazil, using Tradescantia pallida.

The micronucleus (MN) test and analysis of heavy metal biological accumulation in Tradescantia pallida (T. pallida) were bioassays used to assess the genotoxic potential of emissions from a complex of ceramic industries into the atmosphere in a city in Brazil that is considered a national reference source for roof tile production. The ceramic industry emission-exposed T. pallida plants were biomonitored during the dry season, in June, July, and August 2013. In addition to the contaminated monitoring site, a reference site in a peri-urban area was utilized, for comparative purposes. Genotoxicity assessments were determined monthly, while heavy metal bioaccumulation was measured at the end of the total exposure period. The MN frequency was significantly greater in T. pallida plants exposed in the ceramic industry emission monitored area compared to the reference site, and highest MN rates were observed in July and August. With respect to heavy metal bioaccumulation in T. pallida leaves, cadmium (Cd) and chromium (Cr) concentrations were significantly higher in plants at the ceramic industry emission monitoring site. Thus, in relation to the parameters assessed, T. pallida was found to be sensitive to atmospheric contamination by heavy metals attributed to ceramic products emissions generated by the ceramic industry, confirming that this plant species may be employed as a reference organism in biomonitoring studies.

Genotoxicity of Water Contaminants from the Basin of Lake Sevan, Armenia Evaluated by the Comet Assay in Gibel Carp (Carassius auratus gibelio) and Tradescantia Bioassays.

Combination of bioassays and chemical analysis was applied to determine the genotoxic/mutagenic contamination in four different sites of the basin of Lake Sevan in Armenia. Water genotoxicity was evaluated using the single cell gel electrophoresis technique (comet assay) in erythrocytes of gibel carp (Carassius auratus gibelio), Tradescantia micronucleus (Trad-MCN) and Tradescantia stamen hair mutation (Trad-SHM) assays. Significant inter-site differences in the levels of water genotoxicity according to fish and Trad-MCN bioassays have been revealed. Two groups of locations with lower (south-southwest of the village Shorzha and Peninsula of Lake Sevan) and higher (estuaries of Gavaraget and Dzknaget rivers) levels of water genotoxicity were distinguished. Correlation analysis support the hypothesis that the observed genetic alterations in fish and plant may be a manifestation of the effects of water contamination by nitrate ions, Si, Al, Fe, Mn and Cu. Increase of DNA damage in fish also correlated with content of total phosphorus.

Analyses of combined effects of cytostatic drugs on micronucleus formation in the Tradescantia.

Recent experiments showed that 5-fluorouracil (5FU), cisplatin (CDDP), etoposide (ET), and imatinib mesylate (IM), which are currently among the most widely used anticancer drugs, cause damage of the genetic material in higher plants. The aim of the present study was to determine whether mixtures of these drugs cause synergistic or antagonistic effects which may have an impact on their environmental safety. Therefore, the effects of binary mixtures of these anticancer drugs on the induction of micronuclei (MN) which reflect structural and numerical chromosomal aberrations were assessed in Tradescantia tetrads. Synergistic/antagonistic effects were determined by comparison with single exposures that would be equally effective in a reference model of independent action. This comparison was performed at two distinct effect sizes. We found clear evidence for synergisms in combination experiments with IM and antagonism in a high-dose experiment with ET and 5FU. Our findings indicate that IM increases the genotoxic effects of other anticancer drugs. The maximal effects which we found were in the range between 19 and 38 % in the excess of effect sizes predicted under independent action. These effects may have an impact on the overall genotoxic activities of untreated hospital waste waters but not on the environment in general as the predicted environmental concentrations of the studied drugs are several orders of magnitude lower as the levels which are required to cause induction of MN in higher plants.