Use of plant genotoxicity bioassay for the evaluation of efficiency of algal biofilters in bioremediation of toxic industrial effluent

The toxicity and efficacy of an algal-based bioremediation technology were assessed through bioassays for ecological risk of contaminated industrial effluents. The algal bioremoval of heavy metals was evaluated using an in vitro approach. Phytogenotoxicity tests were conducted with Allium cepa and Vicia faba plants to evaluate the genotoxicity of the industrial effluents before and after treatment with different kinds of algal biofilters (BF). Root cells were exposed for 24 h to different dilutions of both raw and treated effluent of a chemical fertilizer factory. Three cytogenetic endpoints were used to assess the mutagenic potencies of the industrial effluent: mitotic inhibition, mitotic chromosome aberrations, and nuclear irregularities in interphase cells. Before algal treatment, the industrial effluent caused strong genotoxic effects represented by severe inhibition in mitotic activity of meristematic cells and high frequency of both chromosome and nucleus abnormalities. After algal treatment, the cytotoxic effects of 30% and 60% concentrations of the treated effluent were comparable to those of 5% and 10% concentrations before treatment, respectively, and the frequency of both chromosome and nuclear abnormalities declined by approximately 50%. Statistical analysis of the data indicates a significant reduction in genotoxicity associated with a remarkable reduction in heavy metal concentrations after bioremediation by algal BF. The Allium and Vicia genotoxicity approach was effective in monitoring bioremediated effluent for toxicity.     

Comet Assay to Assess the Genotoxicity of Persian Walnut (Juglans regia L.) Husks with Statistical Evaluation

The aim of this study was to confirm the utility of the Comet assay as a genotoxicity screening test for evaluating the impact of walnut husk aqueous extract. Phytotoxicity assays using diluted and undiluted walnut husk aqueous extracts were performed on young roots of Raphanus sativus (radish), and the Comet assay was used to evaluate DNA integrity in isolated radish radicle nuclei. The results reveal a dose-dependent accumulation of DNA damage in radish radicles treated with walnut husks water extract and that the Kolmogorov-Smirnov test combined with Johnson SB distribution was the best approach for describing Comet assay data.     

Soil contamination detected using bacterial and plant mutagenicity tests and chemical analyses

Soil contaminants are common in industrialized countries, causing widespread contamination directly of soil and indirectly of ground water and food. Among these pollutants particular attention should be paid to soil mutagens and carcinogens due to their potentially hazardous effects on animal populations and human health. The aim of this research was to evaluate the genotoxicity of contaminated soils by means of an integrated chemical/biological approach, using a short-term bacterial mutagenicity test (Ames test), a plant genotoxicity test (Tradescantia/micronucleus test), and chemical analyses. Soil samples were collected in a highly industrialized area in the Lombardy region, in Northern Italy. Soil samples were extracted with water or with organic solvents. Water extracts of soil samples were tested using the Tradescantia genotoxicity test and organic solvent extracts were analyzed for their polycyclic aromatic hydrocarbon (PAH) concentrations and for their mutagenicity with the Ames test. Heavy metal concentrations were also studied. Some soil samples showed mutagenic activity with the Ames test and clastogenicity with the Tradescantia/micronucleus test. The same soils showed high concentrations of genotoxic PAH and heavy metals.     

Phytoextraction Potential of <Emphasis Type="Italic">Prosopis juliflora</Emphasis> (Sw.) DC. with Specific Reference to Lead and Cadmium

Root and shoot samples of Prosopis juliflora were assessed for their heavy metal content to evaluate the species as a green solution to decontaminate soils contaminated with lead and cadmium. The highest uptake of both the metals was observed in plants from industrial sites. Sites with more anthropogenic disturbance exhibited reduced chlorophyll levels, stunted growth, delayed and shortened reproductive phase. The ratios of lead and cadmium in leaves to lead and cadmium in soil were in the range of 0.62–1.46 and 0.55–1.71, respectively. Strong correlation between the degree of contamination and concentrations of lead and cadmium in plant samples identifies P. juliflora as an effective heavy metal remediator coupled with environmental stress.     

Assessment of hormonal activities and genotoxicity of industrial effluents using in vitro bioassays combined with chemical analysis

Wastewaters from various industries are a main source of the contaminants in aquatic environments. The authors evaluated the hormonal activities (estrogenic/anti-estrogenic activities, androgenic/anti-androgenic activities) and genotoxicity of various effluents from textile and dyeing plants, electronic and electroplate factories, pulp and paper mills, fine chemical factories, and municipal wastewater treatment plants in the Pearl River Delta region by using in vitro bioassays (yeast estrogen screen [YES]; yeast androgen screen [YAS]; and genotoxicity assay [umu/SOS]) combined with chemical analysis. The results demonstrated the presence of estrogenic, anti-estrogenic, and anti-androgenic activity in most industrial effluents, whereas no androgenic activities were detected in all of the effluents. The measured estrogenic activities expressed as estradiol equivalent concentrations (EEQs) ranged from below detection (3 of 26 samples) to 40.7 ng/L, with a mean of 7.33 ng/L in all effluents. A good linear relationship was found between the EEQs measured by YES bioassay and the EEQs calculated from chemical concentrations. These detected estrogenic compounds, such as 4-nonylphenol and estrone, were responsible for the estrogenic activities in the effluents. The genotoxic effects expressed as benzo[a]pyrene equivalent concentrations (BaP EQs) varied between below detection and 88.2 μg/L, with a mean of 8.76 μg/L in all effluents. The target polycyclic aromatic hydrocarbons were minor contributors to the genotoxicity in the effluents, and some nontarget compounds in the effluents were responsible for the measured genotoxicity. In terms of estrogenic activities and genotoxicity, discharge of these effluents could pose high risks to aquatic organisms in the receiving environments.     

Assessment of the Genotoxicity of Endosulfan in Earthworm and White Clover Plants Using the Comet Assay

Endosulfan, as one of the most widely used organochlorine pesticides in the world, has increased the public concern about genotoxicity in soil ecosystems. The comet assay has been widely used in the fields of genetic toxicology and environmental biomonitoring. In the present study we conducted comet assay of endosulfan in earthworm (Eisenia foetida) and white clover (Trifolium repens L.), which are sensitive organisms suitable for acting as a bioindicator for agricultural ecosystems. Earthworms were exposed to endosulfan concentrations of 0.1, 1.0, and 10.0 mg/kg in the soil. White clover roots were immersed in hydroponic pots containing nutrient solutions of different endosulfan concentrations: 0.1, 1.0, and 10.0 mg/L. Tissues from each treatment were collected on the 7th, 14th, 21st, and 28th days of treatment process. Significant effects (p < 0.01) of both concentrations and times of exposure were observed. And endosulfan induced DNA damage in earthworm and white clover nuclei. The comet assay can be used as a reliable tool for early detection of endosulfan.     

Modulatory role of copper on β-naphthoflavone-induced DNA damage in European eel (Anguilla anguilla L.)

Gill and kidney DNA integrity (alkaline unwinding assay) was assessed in Anguilla anguilla exposed for 24-h to copper (Cu: 1 or 2.5 μM), with or without 24-h pre-exposure to a polycyclic aromatic hydrocarbon (PAH)-like compound—β-naphthoflavone (BNF: 2.7 μM). Gill showed DNA integrity loss in all the exposure conditions, reflecting a dual mode of BNF–Cu interaction depending on the metal concentration. Thus, antagonistic or additive effects were observed for BNF+Cu 1 μM or BNF+Cu 2.5 μM, respectively. Kidney showed decreased DNA integrity for single exposures (BNF, Cu 1 μM), whereas sequential exposures displayed higher DNA integrity than BNF alone, revealing a Cu antagonistic effect at both the concentrations. The results also demonstrated that (i) both organs are receptive for Cu inhibitory role against BNF genotoxicity; (ii) kidney is more resistant to Cu individual exposures; and (iii) under multi-pollution conditions genotoxicity cannot be predicted on the basis of individual chemicals responses.     

Comparison of microbial tests for the detection of heavy metal genotoxicity

Heavy metal genotoxicity was evaluated by using different microbial tests. Four genotoxicity assays were employed: the Ames test, the E. coli WP2 test, the Mutatox test detecting mutagenicity, and the SOS assay with E. coli-detecting enzyme induction. All the metals tested (cadmium, chromium, copper, mercury, nickel, and zinc) were detected as genotoxic by the Mutatox and the SOS tests. The Ames test and the E. coli WP2 assay only detected chromium as genotoxic, causing a mutagenic effect. The sensitivity to metals of all the assays used was maintained when they were dissolved in sewage, although there was a slight increase in the sensitivity thresholds.     

Baccharis trimera (Less.) DC as Genotoxicity Indicator of Exposure to Coal and Emissions from a Thermal Power Plant

During coal combustion, hazardous elements are discharged that impair environmental quality. Plant cover is the first available surface for the atmospheric pollutants in terrestrial ecosystems. The aim of this study was to evaluate genotoxicity in the aqueous extract of the native plant, Baccharis trimera, exposed to coal and emissions from a thermal power plant (coal-fired power plant in Candiota, Brazil), correlating seasonality, wind tunnel predominance, and presence of inorganic elements. The presence of inorganic elements in the aerial parts of B. trimera was analyzed by particle-induced X-ray emission (PIXE) spectrometry, and genotoxicity was evaluated by ex vivo comet assay. The genotoxic effects of aqueous extracts of B. trimera from four sites located in the area around power plant were analyzed by comet assay in peripheral human lymphocytes. Winter samples showed greater levels of metals than summer samples. Genotoxicity was detected in B. trimera extracts collected from the region exposed to extraction and burning coal. Extracts from the site impacted by the dominant wind induced more damage to DNA than those from other sites. Based on our data, we can suggest that in winter the inorganic elements from extraction and burning of coal and carried through the wind tunnel were responsible for the genotoxicity observed in aqueous extract of B. trimera.     

Biotests and biosensors in ecotoxicological risk assessment of field soils polluted with zinc, lead, and cadmium

The combined chemical and ecotoxicological hazard evaluation study was conducted on 60 smelter-influenced soils containing 1 to 13, 50 to 653, and 100 to 1,198 mg/kg of Cd, Pb, and Zn, respectively. For these soils (liquid-to-soil ratio = 10), water extractability of Zn, Cd, and Pb was less than 0.19% (median values). Acetic acid (0.11 M) extracted 23, 9.7, and 0.7% of Cd, Zn, and Pb, respectively. Although heavy metal concentrations in the studied soils were high, the toxic effects of water extracts were observed only in few samples and in few biotests (algae Selenastrum capricornutum and metal detector assay). For most of the aquatic test organisms (e.g., crustaceans, photobacteria), the bioavailable concentrations of metals in soil-water extracts were either subtoxic, or the adverse effects were compensated by soil nutrients, etc. However, analysis of the soils with recombinant Cd sensor Bacillus subtilis (pTOO24) showed that about 65% of these apparently subtoxic samples contained bioavailable Cd when analyzed in the suspension assay (detection limit 1.5 mg Cd/kg soil), indicating the desorption of Cd induced by direct contact of bacteria with soil particles. The median bioavailable fraction of Cd (1%) was 23-fold lower than the fraction extracted by acetic acid. The Pb-Cd sensor Staphylococcus aureus (pT0024) detected bioavailable Pb only in the suspensions of five of the most lead-polluted soils (>417 mg Pb/kg): the median bioavailability of Pb was 0.42%. Consequently, the hazard assessment relying on total metal levels in soils should be revised by critical comparison with data obtained from bioassays. Development and use of biosensors (excellent tools for mechanistic studies and signaling hazard already at subtoxic level) should be encouraged.     

River water genotoxicity evaluation using micronucleus assay in fish erythrocytes

The quality of Caí river water (Rio Grande do Sul State) in an area under the influence of a petrochemical complex was studied using the micronucleus assay in erythrocytes from peripheral blood of the fathead minnow Pimephales promelas. This cytogenetic in vivo assay was performed to evaluate the effects of petrochemical effluents on the stream. Organisms were exposed to samples collected at four sites, during an 11-month period. Three different exposure periods were used (7, 14, and 21 days) to evaluate their influence in genotoxic detection. The 14-day exposure period was most effective in detecting genotoxicity in samples from this area. The presence of substances with clastogenic and/or aneugenic potential could be detected at the different sites analyzed. This in vivo assay allowed the detection of genotoxicity in the area studied, indicating the potential for environmental genotoxicity monitoring.     

Beryllium metal I. experimental results on acute oral toxicity, local skin and eye effects, and genotoxicity

The toxicity of soluble metal compounds is often different from that of the parent metal. Since no reliable data on acute toxicity, local effects, and mutagenicity of beryllium metal have ever been generated, beryllium metal powder was tested according to the respective Organisation for Economical Co-Operation and Development (OECD) guidelines. Acute oral toxicity of beryllium metal was investigated in rats and local effects on skin and eye in rabbits. Skin-sensitizing properties were investigated in guinea pigs (maximization method). Basic knowledge about systemic bioavailability is important for the design of genotoxicity tests on poorly soluble substances. Therefore, it was necessary to experimentally compare the capacities of beryllium chloride and beryllium metal to form ions under simulated human lung conditions. Solubility of beryllium metal in artificial lung fluid was low, while solubility in artificial lysosomal fluid was moderate. Beryllium chloride dissolution kinetics were largely different, and thus, metal extracts were used in the in vitro genotoxicity tests. Genotoxicity was investigated in vitro in a bacterial reverse mutagenicity assay, a mammalian cell gene mutation assay, a mammalian cell chromosome aberration assay, and an unscheduled DNA synthesis (UDS) assay. In addition, cell transformation was tested in a Syrian hamster embryo cell assay, and potential inhibition of DNA repair was tested by modification of the UDS assay. Beryllium metal was found not to be mutagenic or clastogenic based on the experimental in vitro results. Furthermore, treatment with beryllium metal extracts did not induce DNA repair synthesis, indicative of no DNA-damaging potential of beryllium metal. A cell-transforming potential and a tendency to inhibit DNA repair when the cell is severely damaged by an external stimulus were observed. Beryllium metal was also found not to be a skin or eye irritant, not to be a skin sensitizer, and not to have relevant acute oral toxic properties.     

Dioxin-Like and Endocrine Disruptive Activity of Traffic-Contaminated Soil Samples

Pollution of surface soils by traffic, especially along major highways, can be a significant issue. Numerous studies have demonstrated traffic to be an important source of particulate matter and gas-phase organic air pollutants that produce many types of deleterious effects. This article brings original information about the presence of contaminants with specific mechanisms of action in traffic-influenced soils as determined by bioanalytical approaches and instrumental analyses. The initial phase of the study aimed to compare contamination of soils near highways with those from reference localities, whereas the second phase of the study investigated the influence of traffic pollution in soils at various distances from highways. For the reference areas, forest soils contained greater concentrations of 2,3,7,8-tetrachlorodibenzo-p-dioxin equivalents (TCDD-EQs; 483 to 2094 pg/g) than did arable soils (96 to 478 pg/g), which represent the relevant reference for the studied soils along highways. The total concentration of TCDD-EQs determined in the in vitro transactivation assay ranged from 225 to 27,700 pg/g in traffic-affected soils. The greatest concentration of TCDD-EQs among the studied sites was observed in soils collected near highway D1, which is the primary thoroughfare in the Czech Republic. The concentrations of TCDD-EQs in roadside soils were the greatest and decreased with increased distance from highways, and this spatial distribution corresponded with the levels of polycyclic aromatic hydrocarbons (PAHs). Soils collected 100 m away from highways in most cases contained concentrations of TCDD-EQs similar to background values. Most TCDD-EQ presence was caused by nonpersistent compounds in soils, with a significant contribution from PAHs as well as other unknown nonpersistent chemicals. Extracts from most soils collected near highways exhibited antiestrogenic and in some cases antiandrogenic activities; for several sites the activity was also detected in soils farther from highways. The presence of TCDD-EQs and antihormonal activity in highway-affected soils points to traffic as a source of polluting compounds having specific effects.     

A plant life-cycle bioassay for contaminated soil,with comparison to other bioassays: Mercury and zinc

Bioassays using rapid-cycling plants allow measurement of multiple endpoints and assessment of impacts on both growth and reproduction. Selections of Brassica rapa develop rapidly in a broad range of soils and are very consistent in production of flower and seed. Their sensitivity to variation in growth conditions was investigated to define the variables that most affect performance. Yield differences between soils were substantial, indicating the need for careful selection and use of control treatments. The sensitivity to contaminants was investigated with applications of mercury (Hg) and zinc (Zn) to three soils. In a sand soil, bloom initiation was slowed by <10 mg Hg kg^–1 soil and <50 mg Zn kg^–1 soil. In contrast, lettuce emergence and earthworm survival were less sensitive to these metals in this soil. Survival of Daphnia magna and the Microtox^® assay in soil extracts were more sensitive to Hg than bloom initiation, but less sensitive to Zn. A similar relationship among the bioassays was observed for two finer-textured soils, although for these, effects were usually apparent only at soil metal concentrations >200 mg kg^–1. Enzyme assays were included for comparison, but were not sensitive to Hg contamination. Rapid-cycling B. rapa selections are suitable for routine bioassays, and are representative of several widely distributed and utilized species.     

Application of the Hydra attenuata assay for identifying developmental hazards among natural waters and wastewaters

This study concerns application of the Hydra attenuata assay to detect the developmental toxicity potential of various aqueous samples. First, the assay was modified for testing aqueous samples because water quality has a major impact on aquatic toxicity testing and the results thus obtained. Ranges of sample pH, salinity (conductivity), and hardness were examined for their adverse effects upon the hydra. Adult hydra were unaffected morphologically by pH 5.5–9.5, and the artificial embryo (“embryo”) developed normally in a pH range of 6.25 to 8.25. For water hardness, the minimal affective concentration was 1000 mg/liter (as CaCO₃) in adults and 625 mg/liter in the embryos; the NOAELs for these were 750 mg/liter in the adult and 250 mg/liter CaCO₃ in the embryo. Salinity in excess of 5 ppt was lethal to adults and embryos, indicating the assay may not be applicable to marine or highly saline samples. Finally, grab samples were tested from rivers in Maryland, Pennsylvania, New Jersey, and Delaware, some of which are impacted by industrial and agricultural activities, as well as several samples of industrial wastewaters from one major facility. The assay functioned normally with these diverse samples and yielded results that can be used in assessing the potential developmental hazard of these materials.     

Assessment of Heavy Metal Pollution in Urban Soils of Havana City,Cuba

Concentrations of Co, Ni, Cu, Zn, Pb and Fe in the top-soils (0–10 cm) from urbanized and un-urbanized areas of Havana city were measured by X-ray fluorescence analysis. The mean Co, Ni, Cu, Zn and Pb contents in the urban topsoil samples (13.9 ± 4.1, 66 ± 26, 101 ± 51, 240 ± 132 and 101 ± 161 mg kg⁻¹, respectively) were compared with mean concentrations for other cities around the world. The results revealed the highest concentrations of metals in topsoil samples from industrial sites. Lowest metal contents were determined in the un-urbanized areas. The comparison with Dutch soil quality guidelines showed a slight contamination with Co, Ni Cu and Zn in all studied sites and with Pb in industrial soils. On the other hand, the metal-to-iron normalisation using Earth crust contents as background showed that soils from urbanized areas in Havana city (industrial sites, parks and school grounds) are moderately enriched with zinc, moderately to severe enriched (city parks and school grounds) and severe enriched (industrial sites) with lead. The values of integrated pollution index (IPI) indicated that industrial soils are middle and high contaminated by heavy metals (1.19 ≤ IPI ≤ 7.54), but enrichment index values (EI) shows that metal concentrations on the studied locations are not above the permissible levels for urban agriculture, except soils from power and metallurgical plants surroundings.     

Toxicity Assessment of Two Soils from Jales Mine (Portugal) Using Plants: Growth and Biochemical Parameters

Contaminants in soils can enter food chains through primary producers. Bioavailable contaminants can induce growth, and reproductive or biochemical changes in plants. To evaluate the bioavailability of heavy metals in two soils from Jales mine surroundings, bioassays with the plants Brassica rapa (RCBr) and Avena sativa were performed. Biochemical parameters (protein and malondialdehyde [MDA] content, and catalase and peroxidase activities) were also measured. The soils had different heavy metal contents: JNC soil contained low heavy metal concentrations, whereas JC soil had high heavy metal contents. Results stressed the difference between species sensitivity, with A. sativa showing no toxicity effects when exposed to both soils. On the other hand, B. rapa presented a decrease in growth parameters when exposed to JNC soil and no changes when exposed to JC soil. A Life Cycle Bioassay confirmed this trend for B. rapa exposed to JNC soil, but also evidenced that JC soil was affecting B. rapa in terms of flower and seed pod production. Biochemical assays showed that plants affected by heavy metals also displayed oxidative stress, with an increase in MDA production, reduction of protein content, and reduction of catalase and peroxidase activities. All bioassays revealed that JNC soil, although with a lower heavy metal content, had a higher bioavailable fraction when compared to JC soil, which consequently increased its toxicity to plants.     

Genotoxicity of 4-nonylphenol and nonylphenol ethoxylate mixtures by the use of Saccharomyces cerevisiae D7 mutation assay and use of this text to evaluate the efficiency of biodegradation treatments

Nonylphenol ethoxylates (NPnEOs, where n is the number of ethoxylic units in the molecule) are non-ionic surfactants widely used for domestic and industrial purposes. 4-Nonylphenol (4-NP), the main product of NPnEO biodegradation, is a toxic xenobiotic compound classified as endocrine disrupter. While numerous studies reported the toxicity and oestrogenic activity of nonylphenols, little is known about the mutagenicity of these compounds. In this paper, the genotoxicity of 4-NP and NPnEO mixtures was evaluated by using the D7 strain of Saccharomyces cerevisiae as experimental model. The same genotoxicity tests were applied to effluents deriving from experimental packed-bed bioreactors, developed for the treatment of NPnEO contaminated wastewater, in order to evaluate the residual genotoxic potential with respect to the influent waste. The target compounds fed to the bioreactors were 4-NP and NPnEO mixtures possessing an average of 5 or 1.5 ethoxylic units (Igepal CO-520 and Igepal CO-210, respectively). The results showed that 4-NP induced significant cytotoxic effect on S. cerevisiae cells at 50 mg/L, as well as mutagenic effects at the lowest tested concentrations (12 and 25 mg/L). 4-NP was the most genotoxic compound among those assayed, followed by Igepal CO-210, whereas Igepal CO-520 did not induce genotoxicity at any of the assayed concentrations. The genotoxic effects of 4-NP on yeast cells disappeared after the treatment of 4-NP artificially contaminated water in the bioreactor. This indicates that the biological treatment is capable of removing not only the pollutant, but also the toxicity associated to the compound and its degradation metabolites. This study represents, to the best of our knowledge, the first report that evaluates the genotoxicity of both 4-NP, NPnEOs and their potential aerobic degradation products on an eukaryotic organism. The obtained results suggest that the S. cerevisiae D7 strain is a very effective model microorganism to study the induction of genotoxic damage by the compounds under study. Moreover, this yeast assay has been proved effective to evaluate the detoxification effect deriving from biotreatment processes.     

The genetic toxicology of organic compounds in natural waters and wastewaters

This review was drawn from the literature describing genotoxic organic compounds in natural water (rivers, lakes, streams) and wastewater, as well as from recent discussions with industrial scientists and environmental regulators. Testing of wastewaters for genotoxicity may become a routine requirement for some industrial wastewater discharge permits, not unlike the more common requirement for routine aquatic toxicity tests. The stimuli for this are concerns that aquatic organisms inhabiting waters impacted by wastewater discharges suffer an increased risk of genetic damage or cancer, and that humans utilizing these waters for recreational and drinking water purposes may suffer similar genetic or carcinogenic risks. Some evidence suggests that neoplasia in aquatic organisms is related to habitat contamination, yet field evaluations fail to substantiate adequately a cause-and-effect relationship. Because aquatic organisms respond like mammals to the same genotoxic compounds, the increased burden of genotoxic compounds to the environment may impact certain endemic species. Wastewater discharges may be one source of genotoxic organic compounds in those impacted areas. With respect to potential human health impacts, evidence is supportive of increased cancer risk to individuals drinking water from surface sources; however, this risk may or may not relate to whether the drinking water source received input of wastewater discharges or known carcinogens. Throughout the published literature reviewed herein, the Salmonella/Ames gene mutation test was widely used to assess genotoxic activity, although studies using indigenous plants and aquatic organisms as in vivo monitors of genotoxic activity exist. No "standard" or frequently followed protocols for sample collection, sample processing, selection of tests or their conduct, or interpretation of data exist for most of the genotoxicity studies reviewed. For the Salmonella/Ames test, the aqueous samples were concentrated usually on XAD resin or by liquid:liquid extraction, and without this concentration step few samples exhibited genotoxic activity. Hence, in most instances, the ambient concentration of the compounds causing this activity is below that which is readily detectable by this test, a finding not new to this review. In contrast, aquatic organisms in laboratory and field studies responded to ambient concentrations of genotoxic compounds, thus alleviating the need for sample concentration. However, there appears to be a reluctance to utilize this information for extrapolation to potential human health effects. Unfortunately, no generally accepted and scientifically validated protocol for preparing aqueous samples for genotoxicity testing exists. Developing such a protocol is necessary before embarking on widespread genotoxicity testing of wastewaters, especially if results are to be used for permit compliance.(ABSTRACT TRUNCATED AT 400 WORDS)     

Evaluation of a microplate assay specific for heavy metal toxicity

A rapid, quantitative microbial assay, which is specific for heavy metal toxicity, has been developed. The assay (MetPLATE) is in a 96-well microtitration plate format and is suitable for determining toxicity characteristics such as median inhibitory concentrations. The sensitivity of MetPLATE to heavy metals [Cu, Zn, Cd, Pb, Hg, Cr(III)] was generally higher than Microtox and was of the same order as or better than Daphnia and fish bioassay. MetPLATE was insensitive to organic compounds at concentrations higher than those found in the environment. Six out of 10 industrial wastewaters or process waters surveyed were toxic. Heavy metal analysis of these waters confirmed the presence of heavy metals in the toxic samples. MetPLATE can be run concurrently with other assays for general toxicity to help determine the nature of chemicals causing toxicity.