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.

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.

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.

Assessing the spatial pattern of a river water quality in southern Brazil by multivariate analysis of biological and chemical indicators / Avaliação do padrão espacial da qualidade da água de um rio no Sul do Brasil por meio da análise multivariada de indicadores biológico e químicos

Abstract This study assessed the genotoxicity and chemical quality of the Rio dos Sinos, southern Brazil. During two years, bimonthly, cuttings of Tradescantia pallida var. purpurea with flower buds were exposed to river water samples from Caraá, Santo Antônio da Patrulha, Taquara and Campo Bom, which are municipalities located in the upper, middle and lower stretches of the Rio dos Sinos basin. Simultaneously, chemical parameters were analyzed, rainfall data were surveyed and negative (distilled water) and positive (0.1% formaldehyde) controls were made. Micronuclei (MCN) frequencies were determined in tetrads of pollen grain mother cells. From the upper stretch toward the lower, there was an increase in the frequency of MCN and in concentrations of chemical parameters. Cadmium, lead, copper, total chromium and zinc were present at the four sites and a concentration gradient was not demonstrated along the river. The multivariate analysis revealed that two principal components exist, which accounted for 62.3% of the observed variances. Although genotoxicity was observed in Santo Antônio da Patrulha, the water presented higher mean values for most of the assessed parameters, in the lower stretch, where urbanization and industrialization are greater. The spatial and temporal pattern of water quality observed reinforces the importance of considering the environmental factors and their effects on organisms in an integrated way in watercourse monitoring programs.

Resumo Este estudo avaliou a genotoxicidade e a qualidade química do Rio dos Sinos, Sul do Brasil. Durante dois anos, com periodicidade bimensal, ramos de Tradescantia pallida var. purpurea com botões florais foram expostos a amostras de água do rio de Caraá, Santo Antônio da Patrulha, Taquara e Campo Bom, municípios localizados nos trechos superior, médio e inferior da Bacia do Rio dos Sinos. Simultaneamente, foram analisados parâmetros químicos, levantados dados de precipitação e realizados controles negativos (água destilada) e positivos (0,1% formaldeído). Frequências de micronúcleos (MCN) foram determinadas em tétrades de células-mãe de grão de pólen. Do trecho superior em direção ao inferior, foi observado aumento da frequência de MCN e na concentração de parâmetros químicos. Cádmio, chumbo, cobre, cromo total e zinco estiveram presentes nos quatro pontos amostrais, sem apresentar um gradiente de concentração ao longo do rio. A análise multivariada demonstrou a existência de dois componentes principais que explicaram 62,3% das variâncias observadas. Embora em Santo Antônio da Patrulha tenha sido observada genotoxicidade, a água do Rio dos Sinos apresentou valores médios superiores para a maioria dos parâmetros avaliados no trecho inferior, onde a urbanização e a industrialização são maiores. O padrão espacial e temporal de qualidade da água observado reforça a importância de considerar os fatores ambientais e seus efeitos nos organismos de forma integrada em programas de monitoramento de cursos hídricos.

Integrated Environmental Assessment of streams in the Sinos River basin in the state of Rio Grande do Sul, Brazil / Avaliação Ambiental Integrada de arroios da bacia do Rio dos Sinos no Estado do Rio Grande do Sul, Brasil

The deterioration of environmental quality in the Sinos River basin is directly associated with the impacts of intense industrialization and urbanization. An integrated environmental assessment (IEA) was conducted in July and September of 2012, in areas along the sources of the EstânciaVelha/Portão, Pampa and Schmidt streams using physical, chemical and biological methods. The water in the three sampling sites was not proper for human consumption, presented a low toxic contamination index (TCI) and mesotrophic characteristics. One site was included in Class 4, and two, in Class 3, according to current legislation. The rapid assessment protocol (RAP) indicated a natural environmental condition for habitat diversity and environmental impact in the three sites. The Tradescantia pallida (Rose) D.R. Hunt var. purpurea Boom biomarker showed water genotoxicity in two of the sites. The integrated diagnosis of water quality in these streams is fundamentally important to ensure the sustainable management of water resources and their multiple uses, as well to estimate their contribution to pollution in this river basin.

A deterioração da qualidade ambiental da bacia do Rio dos Sinos está diretamente associada a impactos oriundos da intensa industrialização e urbanização. A avaliação ambiental integrada (AAI) foi realizada nos meses de julho e setembro de 2012, em áreas junto às nascentes dos arroios Estância Velha/Portão, Pampa e Schmidt por meio de metodologias que utilizam parâmetros físicos, químicos e biológicos. Os três sítios amostrais apresentaram águas impróprias para o consumo humano, baixo índice de contaminação por tóxicos (ICT) e características mesotróficas. Um sítio foi enquadrado na Classe 4 e dois na Classe 3, segundo a legislação vigente. Os resultados da aplicação do protocolo de avaliação rápida (PAR) indicaram uma situação ambiental natural, quanto à diversidade de habitats e aos impactos ambientais, nos três sítios amostrados. O biondicador Tradescantia pallida (Rose) D.R. Hunt var. purpurea Boom evidenciou genotoxicidade da água em dois sítios amostrados. O diagnóstico integrado da qualidade das águas destes arroios é de fundamental importância para assegurar o gerenciamento sustentado dos recursos hídricos e seus múltiplos usos, além de estimar sua contribuição à poluição desta bacia hidrográfica.

Biomonitoring of water genotoxicity in a Conservation Unit in the Sinos River Basin, Southern Brazil, using the Tradescantiamicronucleus bioassay / Biomonitoramento da genotoxicidade da água em uma Unidade de Conservação na Bacia do Rio dos Sinos, Sul do Brasil, utilizando o bioensaio de micronúcleo em Tradescantia

The Tradescantia micronucleus (Trad-MCN) bioassay was used to investigate genotoxicity of water bodies in the Parque Municipal Henrique Luís Roessler (PMHLR), a conservation unit in the city of Novo Hamburgo, Southern Brazil, from November 2010 to October 2011. Every month, cuttings with young inflorescences of Tradescantia pallida var. purpurea were exposed for 24 hours to water collected at three sites in the park: (S1) head of the main stream; (S2) head of a secondary stream; (S3) a point past the junction of the two water bodies in which S1 and S2 were located. As a negative control, cuttings were exposed to distilled water for 24 h every quarter. Micronuclei (MCN) frequency was determined in young tetrads of pollen mother cells and described as MCN/100 tetrads. Rainfall data were also recorded. In nine months at S1 and S2, and in eleven months at S3, micronuclei frequencies were significantly higher than in quarterly controls, in which frequencies varied from 1.19 to 1.62.

During sampling, no significant differences were found in MCN frequencies at S1, which ranged from 2.2 to 3.6. At the other sampling sites, there were significant differences between the months evaluated, and MCN frequencies ranged from 1.3 to 6.5 at S2 and from 2.3 to 5.2 at S3. There were no associations between rainfall and MCN frequencies at the three sampling sites. Tradescantia pallida var. purpureaconfirmed genotoxicity in the water bodies of the PMHLR, even at the head of the streams, which suggests that actions should be promoted to control anthropogenic effects in the streams of this conservation unit.

O bioensaio de micronúcleo em Tradescantia (Trad-MCN) foi utilizado para investigar a genotoxicidade de corpos d'água no Parque Municipal Henrique Luís Roessler (PMHLR), uma Unidade de Conservação no município de Novo Hamburgo, Sul do Brasil, de novembro de 2010 a outubro de 2011. Mensalmente, ramos com inflorescências jovens de Tradescantia pallida var. purpurea foram expostos por 24 horas a amostras de água coletadas em três pontos de cursos d’água no interior do parque: (S1) nascente do principal curso d’água; (S2) nascente de um curso d’agua secundário; (S3) ponto situado após a junção dos corpos d’água em que S1 e S2 estavam localizados. Como controle negativo, ramos foram expostos à água destilada por 24 h, com periodicidade trimestral. Frequências de micronúcleos (MCN) foram determinadas em tétrades jovens de células-mãe de grãos de pólen e expressas como MCN/100 tétrades. Dados de precipitação também foram registrados durante o experimento. Foram observadas frequências de MCN significativamente superiores em nove meses em S1 e S2 e em onze meses em S3 quando comparadas às frequências nos controles trimestrais, que variaram de 1,19 a 1,62. Durante o período de amostragem, não foram encontradas diferenças significativas nas frequências de MCN em S1, que variaram de 2,2 a 3,6. Nos demais pontos amostrais, houve diferenças significativas entre os meses avaliados para as frequências de MCN, que variaram de 1,3 a 6,5, em S2, e de 2,3 a 5,2, em S3. Não foi verificada relação entre a pluviosidade e as frequências de MCN observadas nos três pontos amostrais. Tradescantia pallida var. purpurea indicou haver genotoxicidade nos corpos hídricos do PMHLR, inclusive junto às nascentes, indicando a necessidade de ações que visem ao controle do efeito antropogênico sobre a água nesta unidade de conservação.

Micronucleus assay with tetrad cells of Tradescantia.

The Tradescantia micronucleus assay is being used since almost 50 years for the detection of genotoxins (including carcinogens) in the environment. A large database on the effects of individual compounds and of complex environmental mixtures (soil, air and water) has accumulated. In contrast to other mutagenicity test systems, the effects of low concentrations of heavy metals, radionuclides, certain herbicides and pesticides, and gaseous mutagens can be detected and it is also possible to use the test for in situ biomonitoring studies. 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.

Genotoxicity on Tradescantia pallida var. purpurea plants exposed to urban and rural environments in the metropolitan area of Porto Alegre, southern Brazil / Genotoxicidade em plantas de Tradescantia pallida var. purpurea expostas a ambientes urbano e rural na área metropolitana de Porto Alegre, Região Sul do Brasil

The Trad-MCN bioassay was used to investigate the genotoxicity on Tradescantia pallida var. purpurea plants exposed to variations in the environmental conditions in urban and rural sites in the metropolitan area of Porto Alegre, southern Brazil, over a one-year period. In spring 2009 and in summer, autumn and winter 2010, potted plants of T. pallida var. purpurea were exposed at two sites with different characteristics: the urban area of the municipality of Estância Velha, with leather and footwear industrial activity, and a Site of Special Environmental Interest in the rural area of the municipality of Novo Hamburgo. Other plants comprised the control group and were kept indoors. Frequencies of micronuclei (MCN) were determined in early tetrads of pollen mother cells and expressed as MCN/100 tetrads. Climate data were also registered during the experiment. MCN frequencies in the urban area were significantly higher (up to 8.13) than those found in the rural area (up to 1.26) and in the control group (up to 1.10), which did not differ statistically from each other over the year. The higher MCN frequencies observed in the urban site can be attributed to air pollution, but also may have been influenced by microclimatic and daily thermal variation differences between sites. Higher temperatures recorded in spring and summer may have influenced MCN frequencies observed in the urban site. No clear relation was observed between rainfall and MCN frequencies. Similar and high relative humidity percentages were registered over the period of the study. Considering that the bioindicator plant presents an integrated response to abiotic factors such as pollutants and weather conditions, it can be used as an additional tool that can point to synergistic effects of environmental variables on organisms.

O bioensaio Trad-MCN foi utilizado para investigar a genotoxicidade em plantas de Tradescantia pallida var. purpurea expostas a variações nas condições ambientais, em uma área urbana e uma área rural, na região metropolitana de Porto Alegre, Região Sul do Brasil, durante o período de um ano. Na primavera de 2009 e no verão, no outono e no inverno de 2010, plantas de T. pallida var. purpurea em vasos foram expostas em dois pontos amostrais, com diferentes características: a área urbana do município de Estância Velha, com atividade industrial do setor coureiro calçadista, e uma Área de Especial Interesse Ambiental (AEIA), na zona rural do município de Novo Hamburgo. Outras plantas compuseram o grupo controle e foram mantidas em ambiente interno. Frequências de micronúcleos (MCN) foram determinadas em tétrades jovens de células-mãe de grãos de pólen e expressas como MCN/100 tétrades. Dados climáticos também foram registrados durante o experimento. As frequências de MCN na área urbana foram significantemente superiores (até 8,13) àquelas encontradas na área rural (até 1,26) e no grupo controle (até 1,10), as quais não diferiram estatisticamente entre si ao longo do ano. As maiores frequências de MCN observadas na área urbana podem ser atribuídas à poluição atmosférica, mas também podem ter sido influenciadas por diferenças microclimáticas e de variação térmica diária entre áreas. Temperaturas mais altas registradas na primavera e no verão podem ter influenciado as frequências de MCN observadas na área urbana. Não foi observada uma clara relação entre a pluviosidade e as frequências de MCN. Similares e altas porcentagens de umidade relativa foram registradas ao longo do período de estudo. Considerando‑se que a planta bioindicadora apresenta uma resposta integrada a fatores abióticos, tais como poluentes e condições climáticas, esta pode vir a ser utilizada como uma ferramenta adicional, que pode apontar para efeitos sinérgicos de variáveis ambientais sobre os organismos.

Ability of Allium cepa L. root tips and Tradescantia pallida var. purpurea in N-nitrosodiethylamine genotoxicity and mutagenicity evaluation.

N-nitroso compounds, such as N-nitrosodiethylamine (NDEA), can be formed by the reaction of secondary amines with nitrosating agents, and are suspected to be involved in tumors in humans. NDEA has been considered a weak carcinogen in genotoxic assays probably due to the inefficient nitrosamine activation system that is used and/or to the efficient repair system. In this work, we evaluated the sensibility of Allium cepa L. root tips and Tradescantia stamen hair mutation assay (Trad-SH) using Tradescantia pallida var. purpurea for NDEA (0.1; 0.5; 5 and 25 mM) genotoxicity and mutagenicity induction. Allium cepa L. was treated with different NDEA concentrations for 3h, for 3 consecutive days, including negative control (distilled water) and positive control maleic hydrazide (MH 30 mg/mL). After treatment, the roots were hydrolyzed, squashed, and the mitotic index (MI) and cytological abnormalities were scored. The results revealed a cytostatic effect of NDEA (0.5 and 5mM), showing a significant reduction in the MI. Chromosome stickiness suggests a NDEA toxic effect. T. pallida purpurea did not respond to mutagens with a dose-dependent pattern. In conclusion, our study indicates that the root tips of Allium cepa L. have sensibility to detect NDEA genotoxicity, but not for Trad-SH test.

Ability of Allium cepa L. root tips and Tradescantia pallida var. purpurea in N-nitrosodiethylamine genotoxicity and mutagenicity evaluation

N-nitroso compounds, such as N-nitrosodiethylamine (NDEA), can be formed by the reaction of secundary amines with nitrosating agents, and are suspected to be involved in tumors in humans. NDEA has been considered a weak carcinogen in genotoxic assays probably due to the inefficient nitrosamine activation system that is used and/or to the efficient repair system. In this work, we evaluated the sensibility of Allium cepa L. root tips and Tradescantia stamen hair mutation assay (Trad-SH) using Tradescantia pallida var. purpurea for NDEA (0.1; 0.5; 5 and 25mM) genotoxicity and mutagenicity induction. Allium cepa L. was treated with different NDEA concentrations for 3h, for 3 consecutive days, including negative control (distilled water) and positive control maleic hydrazide (MH 30mg/mL). After treatment, the roots were hydrolyzed, squashed, and the mitotic index (MI) and cytological abnormalities were scored. The results revealed a cytostatic effect of NDEA (0.5 and 5mM), showing a significant reduction in the MI. Chromosome stickiness suggests a NDEA toxic effect. T. pallida purpurea did not respond to mutagens with a dose-dependent pattern. In conclusion, our study indicates that the root tips of Allium cepa L. have sensibility to detect NDEA genotoxicity, but not for Trad-SH test.

Nitrocompostos, como N-nitrosodietilamina (NDEA), podem ser formados pela reação entre uma amina secundária e agentes nitrosantes e são suspeitos de estarem envolvidos na formação de tumores em humanos. NDEA é considerada um carcinógeno fraco e ensaios genotóxicos provavelente pela utilização de um sistema de ativação ineficiente e/ou pela utilização de um eficiente sistema de reparo. Neste trabalho, nós avaliamos a sensibilidade de ensaios com Alliu cepa L. e Tradescantia pallida var. purpurea (Trad-SH) à genotoxicidade e mutagenicidade induzidas por diferentes concentrações de NDEA (0,1; 0,5; 5 e 25mM) por 3h, por 3 dias consecutivos, incluindo controle negativo (água destilada) e controle positivo, hidrazida maleica (MH 30mg/mL). Depois do tratamento, as raízes foram hidrolizadas, esmagadas e o índice mitótico (IM) e anormalidades citológicas foram contadas. Os resultados revelaram um efeito citostático de NDEA (0,5 e 5mM), pela significante redução do IM. Chromosome stickiness sugere um efeito citotóxico de NDEA. T pallida purpurea não respondeu ao mutágeno com um padrão dose dependente. Em conclusão, nossos estudos indicaram que raízes de Allium cepa L. possue sensibilidade na detecção genotóxica de NDEA, mas não para o ensaio Trad-SH.

Assessing the genotoxicity of urban air pollutants using two in situ plant bioassays.

Genotoxicity of urban air has been analysed almost exclusively in airborne particulates. We monitored the genotoxic effects of airborne pollutants in the urban air of Perugia (Central Italy). Two plant bioindicators with different genetic endpoints were used: micronuclei in meiotic pollen mother cells using Tradescantia-micronucleus bioassay (Trad-MCN) and DNA damage in nuclei of Nicotiana tabacum leaves using comet assay (Nicotiana-comet). Buds of Tradescantia clone # 4430 and young N. tabacum cv. Xanthi plants were exposed for 24 h at three sites with different pollution levels. One control site (indoor control) was also used. The two bioassays showed different sensitivities toward urban pollutants: Trad-MCN assay was the most sensitive, but DNA damage in N. tabacum showed a better correlation with the pollutant concentrations. In situ biomonitoring of airborne genotoxins using higher plants combined with chemical analysis is thus recommended for characterizing genotoxicity of urban air.

Association between micronuclei frequency in pollen mother cells of Tradescantia and mortality due to cancer and cardiovascular diseases: a preliminary study in Sao José dos Campos, Brazil.

The present study was designed to explore the correlation between the frequency of micronuclei in Trad-MN, measured across 28 biomonitoring stations during the period comprised between 11 of May and 2 of October, 2006, and adjusted mortality rates due to cardiovascular, respiratory diseases and cancer in Sao José dos Campos, Brazil, an area with different sources of air pollution. For controlling purposes, mortality rate due to gastrointestinal diseases (an event less prone to be affected by air pollution) was also considered in the analysis. Spatial distribution of micronuclei frequency was determined using average interpolation. The association between health estimators and micronuclei frequency was determined by measures of Pearson's correlation. Higher frequencies of micronuclei were detected in areas with high traffic and close to a petrochemical pole. Significant associations were detected between micronuclei frequency and adjusted mortality rate due to cardiovascular diseases (r=0.841, p=0.036) and cancer (r=0.890, p=0.018). The association between mortality due to chronic obstructive pulmonary diseases was positive but did not reach statistical significance (r=0.640, p=0.172), probably because of the small number of events. Gastrointestinal mortality did not exhibit significant association with micronuclei frequency. Because the small number of observations and the nature of an ecological study, the present findings must be considered with caution and considered as preliminary. Further studies, performed in different conditions of contamination and climate should be done before considering Trad-MN in the evaluation of human health risk imposed by air pollutants.

Evaluation of chlorite and chlorate genotoxicity using plant bioassays and in vitro DNA damage tests.

In the last few years chlorine dioxide has been increasingly used for disinfecting drinking water in many countries. Although it does not react with humic substances, chlorine dioxide added to water is reduced primarily to chlorite and chlorate ions, compounds that are under investigation for their potential adverse effects on human health. The aim of this research was to study the genotoxicity of chlorite and chlorate and their mixtures. The end-points included two plant tests (chromosomal aberration test in Allium cepa and micronucleus assay in Tradescantia, carried out at different times of exposure) and two genotoxicity tests in human HepG2 cells (comet assay and cytokinesis-blocked micronucleus test). Preliminary toxicity tests were carried out for both plant and HepG2 assays. The results showed that chlorite and chlorate are able to induce chromosomal damage to plant systems, particularly chromosomal aberrations in A. cepa root tip cells, even at concentrations lower than the limit established by Italian normative law and WHO guidelines. In HepG2 cells increased DNA damage was only observed for chlorate at the lowest concentration. No increase in micronuclei frequency was detected in any of the samples tested in human HepG2 cells.

Evaluation of toxic and genotoxic effects of low-level 137Cs ionising radiation on plants.

The aim of this study was to evaluate the impact of low internal exposure to 137Cs on L. sativum meristem cells and Tradescantia stamen hair cells. It also compared the impact of 137Cs internal and external irradiation of similar level on the plant seed germination and root growth. Compared to control, the tested internal (0.0007 mGy to 0.7 mGy) and external (0.04 mGy to 5.5 mGy) 137Cs ionising radiation doses stimulated the elongation of L. sativum roots by 11% to 12% and 24% to 33%, respectively. Internal 137Cs exposure (0.0003 mGy to 0.5 mGy) for 14 days caused 1.2% to 1.6% of somatic mutations and 19% to 87% of non-viable stamen hair in Tradescantia.

Tradescantia micronucleus test indicates genotoxic potential of traffic emissions in European cities.

Urban atmospheres contain complex mixtures of air pollutants including mutagenic and carcinogenic substances such as benzene, diesel soot, heavy metals and polycyclic aromatic hydrocarbons. In the frame of a European network for the assessment of air quality by the use of bioindicator plants, the Tradescantia micronucleus (Trad-MCN) test was applied to examine the genotoxicity of urban air pollution. Cuttings of Tradescantia clone #4430 were exposed to ambient air at 65 monitoring sites in 10 conurbations employing a standardised methodology. The tests revealed an elevated genotoxic potential mainly at those urban sites which were exposed to severe car traffic emissions. This bioassay proved to be a suitable tool to detect local 'hot spots' of mutagenic air pollution in urban areas. For its use in routine monitoring programmes, however, further standardisation of cultivation and exposure techniques is recommended in order to reduce the variability of results due to varying environmental conditions.

Genotoxic effects of extremely low frequency (ELF) magnetic fields (MF) evaluated by the Tradescantia-micronucleus assay.

Extremely low frequency (ELF) electric fields (EF) and magnetic fields (MF) are generated during the production, transmission, and use of electrical energy. Although epidemiology studies suggest that there is a cancer risk associated with exposure to ELF-MF, short-term genotoxicity assays with bacteria and mammalian cells have produced inconsistent results. In the present study, we investigated the possible genotoxicity of ELF-MF by using the Tradescantia-micronucleus (Trad-MN) assay, a sensitive, reproducible, well-standardized assay for genotoxicity testing. A 50 Hz ELF-MF was generated by a laboratory exposure system consisting of a pair of parallel coils in a Helmholtz configuration. Exposure of Tradescantia (clone # 4430) inflorescences to the ELF-MF, at a flux density (B) corresponding to 1 mT, for 1, 6, and 24 h resulted in a time-dependent increase in MN frequency. The results indicate that a 50 Hz MF of 1 mT field strength is genotoxic in the Trad-MN bioassay and suggest that this assay may be suitable as a biomonitor for detecting the genotoxicity of ELF-MF in the field.

Assessment of the genotoxicity of 137Cs radiation using Vicia-micronucleus, Tradescantia-micronucleus and Tradescantia-stamen-hair mutation bioassays.

Since the middle of the 20th century, ionizing radiations from radioactive isotopes including 137Cs have been investigated to determine their genotoxic impact on living organisms. The present study was designed to compare the effectiveness of three plant bioassays to assess DNA damage induced by low doses of 137Cs: Vicia-micronucleus test (Vicia-MCN), Tradescantia-micronucleus test (Trad-MCN) and Tradescantia-stamen-hair mutation test (Trad-SH) were used. Vicia faba (broad bean) and Tradescantia clone 4430 (spiderwort) were exposed to 137Cs according to different scenarios: external and internal (contamination) irradiations. Experiments were conducted with various levels of radioactivity in solution or in soil, using solid or liquid 137Cs sources. The three bioassays showed different sensitivities to the treatments. Trad-MCN appeared to be the most sensitive test (significative response from 1.5 kBq/200 ml after 30 h of contamination). Moreover, at comparable doses, internal irradiations led to larger effects for the three bioassays. These bioassays are effective tests for assessing the genotoxic effects of radioactive 137Cs pollution.

The study of the effects of low-level exposure to ionizing radiation using a bio-indicator system.

The object of the present study was to apply the high sensitivity of a botanical mutagenicity test (Tradescantia stamen-hair mutation bioassay), in "in situ" bioassays to determine the responses induced by the exposure to low levels of ionizing radiation. Mutagenesis was evaluated during 28 days, in an environment that presented gamma radiation exposure rates of 1.6 (control), 25.0 and 75.0 microR min(-1). From the results we observed that there was no linear response throughout the exposure time when compared mutagenic events and gamma radiation exposure rates were compared. The results obtained in the second week of exposure of Tradescantia showed that after exposure to 25.0 and 75.0 microR min(-1) there was a significant increase (p<0.05) in the mutation levels in Tradescantia stamen-hair. In the third and fourth week after exposure 25.0microR min(-1), the plants demonstrated a mutation rate that was not significantly different from the control (p>0.05).

Comparative study on sensitivity of higher plants and fish to heavy fuel oil.

Laboratory tests were conducted on higher plants [garden cress (Lepidium sativum), great duckweed (Spirodela polyrrhiza), and Tradescantia clone BNL 02] and fish [rainbow trout (Oncorhynchus mykiss) at all stages of development: eggs, larvae and adults] to estimate their sensitivity to heavy fuel oil (HFO). A number of biological indices (survival, growth, and physiological and morphological parameters) as well as the genotoxic impact (Tradescantia) of HFO was evaluated by acute and chronic toxicity tests. Fish were found to be more sensitive to the toxic effect of HFO than were higher plants. EC(50) values obtained for higher plants ranged from 8.7 g/L (L. sativum) to 19.8 g/L (Tradescantia), and maximum-acceptable-toxicant concentration (MATC) values ranged from 0.1 to 1.0 g/L of total HFO for L. sativum and Tradescantia, respectively. The 96-h LC(50) values ranged from 0.33 g/L, for larvae, to 2.97 g/L, for adult fish, and the MATC value for fish was found to be equal to 0.0042 g/L of total HFO. To evaluate and predict the ecological risk of the overall effects of oil spills, studies should be performed using a set of acute and chronic bioassays that include test species of different phylogenetic levels with the most sensitive morphological, physiological, and genotoxic indices.

Genotoxicity of surface water treated with different disinfectants using in situ plant tests.

Disinfection of surface drinking water, in particular water chlorination, results in many by-products with potential genotoxic and/or carcinogenic activity. In the present study, we evaluated the genotoxicity of surface water after treatment with different disinfectants by means of in situ plant genotoxicity assays (micronucleus and chromosomal aberration tests) which can detect both clastogenic and aneugenic effects. The study was carried out at a pilot plant using lake water after sedimentation and filtration. This water supplied four stainless steel basins: three basins were disinfected with sodium hypochlorite, chlorine dioxide, and peracetic acid and the fourth basin containing untreated lake water was used as a control. Plants were exposed in situ in the basins. The study was carried out using water collected in different seasons over a period of about 1 year in order to assess the treatments in different physical and chemical lake water conditions. The micronucleus test in root cells of Vicia faba (Vicia faba/MCN test) revealed genotoxicity in many samples of disinfected water. The micronucleus test in Tradescantia pollen cells and the chromosome aberration test in root cells of Allium cepa showed genotoxic effects only in some disinfected samples, but also revealed genotoxicity in raw water. The results of the study indicated that the Vicia faba/MCN test was the most sensitive plant assay for disinfected water and that peracetic acid disinfection produced similar or lower genotoxicity than sodium hypochlorite or chlorine dioxide treatment.