Assessment of toxicity reduction after metal removal in bioleached sewage sludge

Sewage sludge can be applied to land to supply and recycle organic matter and nutrients. Trace elements in sludge, however, may accumulate in the soil with repeated sludge applications. Reducing metal content may therefore reduce the adverse effects of sludge application. The objective of this study was to evaluate the efficiency of bioleaching technology in reducing metal content and toxicity as measured by a battery of terrestrial and liquid-phase bioassays. Sludge-soil mixtures simulating the application of sludge to land were tested by means of terrestrial bioassays, barley (Hordeum vulgare L.) seed germination (5 d) and sprout growth (14 d), lettuce (Lactuca sativa) seed germination (5 d), and worm (Eisenia andrei) mortality (14 d). Liquid-phase bioassays, Microtox (Vibrio fischeri, 15 min), lettuce root elongation (L. sativa, 5 d), cladoceran mortality (Daphnia magna, 48 h), and SOS Chromotest (Escherichia coli) were used after elutriation of the sludge. Comparison of the bioassay results (except for D. magna) before and after treatment demonstrated that this bioleaching process reduced both sludge toxicity and metal content. In addition, lower Cu and Zn concentrations found in barley sprouts following treatment supported the assumption that the bioleaching process, by decreasing metal content and bioavailability, reduced sewage sludge toxicity. This study also emphasized the interest of using ecotoxicological bioassays for testing biosolids. In particular, the terrestrial bioassays after simulation of land application and the Microtox test after sludge elutriation proved to be the most appropriate procedures.     

Toxicity of benzotriazole and benzotriazole derivatives to three aquatic species

Benzotriazole and its derivatives comprise an important class of corrosion inhibitors, typically used as trace additives in industrial chemical mixtures such as coolants, deicers, surface coatings, cutting fluids, and hydraulic fluids. Recent studies have shown that benzotriazole derivatives are a major component of aircraft deicing fluids (ADFs) responsible for toxicity to bacteria (Microtox). Our current research compared the toxicity of benzotriazole (BT), two methylbenzotriazole (MeBT) isomers, and butylbenzotriazole (BBT). Acute toxicity assays were used to model the response of three common test organisms: Microtox bacteria (Vibrio fischeri), fathead minnow (Pimephales promelas) and water flea (Ceriodaphnia dubia). The response of all the three organisms varied over two orders of magnitude among all compounds. Vibrio fischeri was more sensitive than either C. dubia or P. promelas to all the test materials, while C. dubia was less sensitive than P. promelas. The response of test organisms to unmethylated benzotriazole and 4-methylbenzotriazole was similar, whereas 5-methylbenzotriazole was more toxic than either of these two compounds. BBT was the most toxic benzotriazole derivative tested, inducing acute toxicity at a concentration of < or = 3.3 mg/l to all organisms.     

Use of highly sensitive sublethal stress responses in the social amoeba Dictyostelium discoideum for an assessment of freshwater quality

In this work, the sensitivity of a battery of tests on the social amoeba Dictyostelium discoideum has been assessed within a freshwater toxicity study. The results obtained from the evaluation of survival and replication rate of D. discoideum were compared to those derived with a series of widely used tests for freshwater toxicity assessment, i. e. bioassays using Vibrio fischeri, Daphnia magna and Pseudokirchneriella subcapitata. The effects on sublethal endpoints, i.e. lysosomal membrane stability (LMS) and endocytotic rate, were analysed in conjunction with high-level endpoints to verify the potential to make a typical bioassay more sensitive. The field ecotoxicological investigation employing D. discoideum is part of a monitoring study assessing environmental quality of the Bormida River (Italy), subjected until recently to a chronic industrial pollution. The survey was carried out at several stations (upstream and downstream of a chemical factory outlet) in two different periods. In 2002, the results of chemical analyses performed on river water indicated no contamination. The ecotoxicological data obtained in this period showed that no evidence of biological effects was observed using V. fischeri and D. magna bioassays. In spite of the previous classical acute toxicity tests, significant differences in cell viability of D. discoideum were found. By analysing the effects measured on LMS and endocytotic rate, more relevant changes were observed for these sublethal stress biomarkers compared to survival. The chronic toxicity data showed significant changes in cell growth both of P. subcapitata and D. discoideum. Nevertheless, more sensitive and rapid responses were obtained when assessing the effects of exposure on D. discoideum. The chemical and ecotoxicological data obtained in 2006 indicated a full recovery of the quality of the river water (neither contamination nor toxicity found). Altogether, the results reported in this study underline that the use of a battery of biomarkers in conjunction with high-level endpoints may help follow the pollutant-induced stress syndrome in the organisms from early sublethal effects to starting mortality.     

Responses in sediment bioassays used in the Netherlands: can observed toxicity be explained by routinely monitored priority pollutants?

In order to identify the cause of toxicity in sediments and suspended matter, a large number of samples with different degrees of contamination was taken at various locations in The Netherlands. Standard acute bioassays were carried out with the bacterium Vibrio fischeri, the rotifer Brachionus calyciflorus and the anostracan Thamnocephalus platyurus. Chronic standard tests were performed using the water flea Daphnia magna and larvae of the midge Chironomus riparius. Some novel bioassays were performed as well. Most toxic effects observed in standard bioassays with sediments from polluted sediments (class 3 and 4 on a scale of 0-4 according to the Dutch criteria) could be partly explained by toxic concentrations of known persistent priority pollutants, mainly heavy metals and occasionally polycyclic aromatic hydrocarbons. In some of the samples, ammonia toxicity was a confounding factor during testing. Suspended matter from the Meuse river at Eijsden, which may be considered as 'new' sediment (pollution class 2), was moderately to highly toxic in almost all bioassays. This could have been associated with a combination of heavy metals, PAHs and ammonia. At two locations from the Lake IJssel area with no apparent persistent pollution, moderate and strong effects were nonetheless observed in invertebrate tests. This might have been due to agricultural run-off of pesticides, which are not routinely measured in sediments. A few effects on V. fischeri in canals and a small stream could not be explained with standard chemical analysis, but seemed associated with the outlets of sewage water treatment plants and industrial effluents. Additional chemical analysis of pore water samples from five selected sediments yielded more identified substances such as phtalates, decanes, cosanes and fragrances, but it was estimated that their contribution to the effects observed on V. fischeri, D. magna and C. riparius was negligible.     

Biomonitoring of the genotoxic potential of aqueous extracts of soils and bottom ash resulting from municipal solid waste incineration, using the comet and micronucleus tests on amphibian (Xenopus laevis) larvae and bacterial assays (Mutatox and Ames tests)

The management of contaminated soils and wastes is a matter of considerable human concern. The present study evaluates the genotoxic potential of aqueous extracts of two soils (leachates) and of bottom ash resulting from municipal solid waste incineration (MSWIBA percolate), using amphibian larvae (Xenopus laevis). Soil A was contaminated by residues of solvents and metals and Soil B by polycyclic aromatic hydrocarbons and metals. MSWIBA was predominantly contaminated by metals. Two genotoxic endpoints were analysed in circulating erythrocytes taken from larvae: clastogenic and/or aneugenic effects (micronucleus induction) after 12 days of exposure and DNA-strand-breaking potency (comet assay) after 1 and 12 days of exposure. In addition, in vitro bacterial assays (Mutatox and Ames tests) were carried out and the results were compared with those of the amphibian test. Physicochemical analyses were also taken into account. Results obtained with the amphibians established the genotoxicity of the aqueous extracts and the comet assay revealed that they were genotoxic from the first day of exposure. The latter test could thus be considered as a genotoxicity-screening tool. Although genotoxicity persisted after 12 days' exposure, DNA damage decreased overall between days 1 and 12 in the MSWIBA percolate, in contrast to the soil leachates. Bacterial tests detected genotoxicity only for the leachate of soil A (Mutatox). The results confirm the ecotoxicological relevance of the amphibian model and underscore the importance of bioassays, as a complement to physico-chemical data, for risk evaluation.     

Use of multidimensional scaling in the selection of wastewater toxicity test battery components

In aquatic toxicity testing, no single test species is sensitive to all toxicants. Therefore, test batteries consisting of several individual assays are becoming more common. The organisms in a test battery should be representative of the entire system of interest. The results of the assays should be complementary to other components in the test battery to avoid redundancy. With the aid of multidimensional scaling (MDS), a multivariate statistical method, we examined the toxicity data of five bioassays (the continuous Shk1, Polytox, activated sludge respiration inhibition, Nitrosomonas, and Tetrahymena assays) that could serve as test battery components for the assessment of wastewater toxicity to activated sludge. MDS mapped the five assays into a two-dimensional space and showed that the Nitrosomonas assay should be included in test batteries plus one of the remaining four assays for assessing wastewater toxicity to activated sludge.     

Algae and crustaceans as indicators of bioactivity of industrial wastes

Freshwater (Selenastrum capricornutum) and marine (Skeletonema costatum) algae were exposed to liquid wastes from 10 industrial sites in laboratory bioassays. All wastes affected algal growth, either by stimulation only or by stimulation at low concentrations and inhibition at high concentrations. Generally, S. capricornutum and Sk. costatum responded similarly to each waste: SC20's (concentration that stimulated growth by 20%) were between 0.01 and 20.0% waste; EC50's (concentration that inhibited growth by 50%), between 5.1 and 85.5% waste. Since toxicity to S. capricornutum was usually lost by the sixth or seventh day of exposure in all wastes except one, it is recommended that algal tests be carried out for 4 days. Both algal species were more sensitive to the wastes than were Daphnia magna (freshwater) and Mysidopsis bahia (marine). Only three wastes were toxic to D. magna and two were toxic to M. bahia. SC20 and EC50 values are used to calculate the 7-day, 10-year flow rate of the receiving stream required for dilution of effluents to non-toxic concentrations.     

Quality assessment of bed sediments of the Po River (Italy)

Comprehensive and contemporary evaluations of physical, chemical and toxicological endpoints have been performed on bed sediments of the Po River, the major Italian watercourse. Two extensive sampling campaigns were conducted in summer and winter low-flow conditions. Composite sediment samples were collected from ten reaches of the main river: the first was located in the upper region (ambient control), and the others downstream of the confluences of nine principal tributaries. The two sampling programs were paralleled by contemporary investigations on the macroinvertebrate community. The particle-size composition along the Po River showed a relatively uniform distribution of fine sand, a progressive downstream decrease of coarse sands and a corresponding increase of fine materials. The levels of polychlorinated biphenyls (PCB), polycyclic aromatic hydrocarbons (PAH), extractable organo halides (EOX), Cd, Cr, Cu, Hg, Ni, Pb and Zn were determined in sediment fine particles (< 63 microm), and showed marked changes across the ten river reaches. Their longitudinal trends, as those of organic carbon and total nitrogen, were very similar and largely independent of the survey season. Sediment quality benchmarks were used to evaluate sediment chemistry, and, although the overall level of contamination was from moderate to low, the reaches located downstream of the tributaries Dora Riparia, Dora Baltea, Lambro and Oglio were considered to be at risk. Sediments were tested for toxicity on Oncorhynchus mykiss, Ceriodaphnia dubia, Raphidocelis subcapitata and Vibrio fischeri. The toxicity tests were conducted both with sediment extracts and whole samples. Sediment extracts showed toxic potentials that were consistent with the spatial distribution of contaminants. Whole-sediment toxicity showed moderate/low effects which also included false positives and negatives. Alterations of the macroinvertebrate community were found for many kilometers downstream of Dora Riparia, and with a seasonal dependence, also in other reaches of the Italian river. Principal component analysis (PCA) was used to describe the longitudinal and temporal changes of the Po River, and allowed the selection of the most useful and discriminating indicators.     

Chemical and ecotoxicological assessment of polycyclic aromatic hydrocarbon--contaminated sediments of the Niger Delta, Southern Nigeria

The extent of environmental contamination and sources of polycyclic aromatic hydrocarbons (PAHs) compounds to sediments of the Niger Delta, Nigeria were assessed using combined chemical analysis and toxicity bioassay techniques. Concentrations of two- to six-ring PAHs of molecular mass 128-278 and toxicity to Vibrio fischeri and Lemna minor are considered in this investigation. Levels of the sum of the 16 USEPA priority pollutant PAHs varied from 20.7 to 72.1 ng/g dry weight. Sediment PAH levels were highest in samples collected from Delta Steel located at the outskirts of Warri, and Quality control centre, Ughelli West; with total PAH concentrations of 72.1 and 67.5 ng/g dry weight, respectively. The overall levels of PAHs in this study are low compared to other regions and reveal moderate PAHs pollution in the sediments of the Niger Delta. Two- and three-ring aromatic hydrocarbons predominated in almost all the sediments, which indicate a petrogenic origin. The sediment total PAH (PAHtot) concentration, normalized to organic carbon content (OC), ranged from 120.2 to 1.99 ng PAHtot/mg OC; and showed distinctively that the sedimentary organic matter of the sample from Delta Steel is highly contaminated with PAHs, and had a value of 120.2 ng PAHtot/mg OC. The toxicity bioassays indicated that the sample collected from Warri Refinery Area (SDWRR) was the most toxic to V. fischeri, with an EC50 value of 0.45 mg sediment equiv./mL test medium; and samples from Ogunu (SDOGN) and Warri Refinery area (SDWRR) showed high toxicity to L. minor, with percent inhibitions of 42.6% and 33.67%, respectively, after 7 days of exposure. The total PAH concentrations showed no correlation with toxicity bioassays, and thereby implied that chemical analysis of PAHs cannot be an indicator of sediment toxicity.     

膨胀剂增强地下混凝土结构抗裂效应实测分析

以实际工程为背景,通过比较搅拌站试件试验及实际施工测试结果,分析了混凝土膨胀剂增强地下混凝土结构的膨胀抗裂效应。试件试验结果表明,掺有膨胀剂混凝土的限制膨胀率随时间变化呈现明显规律性,前3 d迅速增长并于第14 d达到峰值,随后随着时间的增加略微降低,第28 d的限制膨胀率基本保持在0.030%。现场实测的混凝土限制膨胀率随时间发展规律与试件试验相同,但其峰值相对较小。现场实测的混凝土膨胀应变相比混凝土限制膨胀率更早达到峰值,但趋势大致相同。掺有膨胀剂的后浇带及后浇式膨胀加强带能有效提高结构混凝土的膨胀应变,增强抗裂效应,距离膨胀加强带和墙体越近膨胀应变值越大。     

Biodegradability of meat industry wastes under anaerobic and aerobic conditions

In this study the biodegradation kinetics and the different biologically degradable fractions (readily, slowly and inert fractions) of the organic wastes generated in a meat industry have been estimated under both anaerobic and aerobic conditions. Three of these could be degraded under both conditions, whereas one (pig/cow waste slurries) could only be aerobically degraded since the high ammonia concentration caused inhibition in anaerobic experiments. Mathematical models for anaerobic and aerobic degradations were used to estimate the readily (S(S)) and slowly (X) biodegradable fractions of the meat industry wastes. Using these models, a good agreement was observed between the calculated S(S) fractions for each waste under anaerobic and aerobic conditions, while the calculated X fractions in the aerobic tests were lower than in the anaerobic tests. The experiments, in combination with the modelling, showed that aerobic respirometric test may be used for predicting readily biodegradable fractions under both anaerobic and aerobic conditions provided that ammonia concentration is not significantly high. Thus, for complex organic wastes the length of experiments for estimating biodegradable fractions may be considerably reduced (in this case by 10-12 days).     

Effects of aqueous, dietary and combined exposures of cadmium to Ceriodaphnia dubia

While effects of water-borne metal exposures on freshwater animals have been well documented, the effect of dietary metal exposure is less understood but is gaining importance. However, little attention has been given to the importance of combining both exposure pathways. In this study, we compared effects of aqueous ('water only'), dietary ('food only') and combined ('water+food') exposures of cadmium to the freshwater cladocerans, Ceriodaphnia dubia. Major test endpoints included survival, feeding rate and reproduction. The C. dubia three-brood reproduction tests were conducted according to the United States Environmental Protection Agency (U.S. EPA) methods. Three exposure scenarios were used: aqueous, dietary, and combined aqueous and dietary exposures. Results showed that all three exposures affected survival, feeding rate and reproduction of C. dubia. Interestingly, combined exposure showed contribution effects of aqueous and dietary exposures. Lower cadmium concentrations were needed in combined exposure to produce effects as compared to higher concentrations in aqueous or dietary exposure alone. These results demonstrated the potential importance of dietary and combined exposures for consideration of cadmium regulation and risk assessment of metals.     

Monitoring the biological activity of micropollutants during advanced wastewater treatment with ozonation and activated carbon filtration

A bioanalytical test battery was used to monitor the removal efficiency of organic micropollutants during advanced wastewater treatment in the South Caboolture Water Reclamation Plant, Queensland, Australia. This plant treats effluent from a conventional sewage treatment plant for industrial water reuse. The aqueous samples were enriched using solid-phase extraction to separate some organic micropollutants of interest from metals, nutrients and matrix components. The bioassays were chosen to provide information on groups of chemicals with a common mode of toxic action. Therefore they can be considered as sum indicators to detect certain relevant groups of chemicals, not as the most ecologically or human health relevant endpoints. The baseline toxicity was quantified with the bioluminescence inhibition test using the marine bacterium Vibrio fischeri. The specific modes of toxic action that were targeted with five additional bioassays included aspects of estrogenicity, dioxin-like activity, genotoxicity, neurotoxicity, and phytotoxicity. While the accompanying publication discusses the treatment steps in more detail by drawing from the results of chemical analysis as well as the bioanalytical results, here we focus on the applicability and limitations of using bioassays for the purpose of determining the treatment efficacy of advanced water treatment and for water quality assessment in general. Results are reported in toxic equivalent concentrations (TEQ), that is, the concentration of a reference compound required to elicit the same response as the unknown and unidentified mixture of micropollutants actually present. TEQ proved to be useful and easily communicable despite some limitations and uncertainties in their derivation based on the mixture toxicity theory. The results obtained were reproducible, robust and sensitive. The TEQ in the influent ranged in the same order of magnitude as typically seen in effluents of conventional sewage treatment plants. In the initial steps of the treatment chain, no significant degradation of micropollutants was observed, and the high levels of dissolved organic carbon probably affected the outcome of the bioassays. The steps of coagulation/flocculation/dissolved air flotation/sand filtration and ozonation decreased the effect-based micropollutant burden significantly.     

Evaluation of the genotoxic and teratogenic potential of a municipal sludge and sludge-amended soil using the amphibian Xenopus laevis and the tobacco: Nicotiana tabacum L. var. xanthi Dulieu

The toxic, genotoxic and teratogenic potential of a municipal sewage sludge was assessed using the micronucleus assay on the larvae of the amphibian Xenopus laevis and with the tobacco somatic mutation test using the yellow-green xanthi Dulieu mutant a(1)(+)/a(1) a(2)(+)/a(2). The teratogenic potential was assessed by means of the Frog Embryo Teratogenesis Assay-Xenopus (FETAX). Various doses of the pasty sludge added to a crop soil were tested using the three bioassays. The test systems were performed either directly with sludge or sludge-amended soil samples (plant model) or with aqueous extracts (aquatic animal model). Using the tobacco model, we found no mutagenic impact of the soil amended with the sludge, perhaps because the clay-like nature of the soil, with its high adsorption capacity, may have prevented the contaminants from reaching the target. All leachates of amended soils produced a significant size reduction in Xenopus embryos. Depending on the soil/sludge ratio, some leachates were found to be genotoxic but were never teratogenic. This battery of in vivo test systems enabled us to estimate the global long-term effects under agricultural conditions with various genetic endpoints on ecologically relevant organisms characteristic of the aquatic and terrestrial compartments.     

Potential of olive mill waste and compost as biobased pesticides against weeds, fungi, and nematodes

The phytotoxic and antimicrobial properties of olive mill wastes have been widely investigated and demonstrated over the past decade. However, their potential utilization as biodegradable pesticides against plant pathogens is still poorly understood. In this study, a series of laboratory bioassays was designed to test the inhibitory effects of sterile water extracts of two-phase olive mill waste (TPOMW) and TPOMW composts with different degrees of stabilization on several different plant pathogens. Fungicidal properties of TPOMW extracts, assayed in a microwell assay format, showed that the growth of Phytophthora capsici was consistently and strongly inhibited by all TPOMW extracts diluted 1:10 (w:v). In contrast, suppression of Pythium ultimum and Botrytis cinerea by the extracts was not as strong and depended on the specific TPOMW sample. Mature compost inhibited P. capsici and B. cinerea at dilutions as great as 1:50, w:v. Neither TPOMW nor TPOMW compost extracts were able to inhibit the growth of the basidiomycete root rot agent Rhizoctonia solani. In addition, studies were conducted on the allelopathic effects of TPOMW extracts on seed germination of four highly invasive and globally distributed weeds (Amaranthus retroflexus, Solanum nigrum, Chenopodium album and Sorghum halepense). Both the TPOMW and immature TPOMW compost extracts substantially inhibited germination of A. retroflexus and S. nigrum, whereas mature composts extracts only partially reduced the germination of S. nigrum. Finally, TPOMW extracts strongly inhibited egg hatch and second-stage juvenile (J2) motility of the root-knot nematode Meloidogyne incognita. However, only higher concentrations of stage-one and stage-two TPOMW compost extracts exerted a suppressive effect on both J2 motility and on egg hatch. The study shows the high potential of naturally occurring chemicals present in TPOMW and TPOMW composts that should be further investigated as bio-pesticides for their use in sustainable agricultural systems.     

Toxicity to Daphnia magna and Vibrio fischeri of Kraft bleach plant effluents treated by catalytic wet-air oxidation

Two Kraft-pulp bleaching effluents from a sequence of treatments which include chlorine dioxide and caustic soda were treated by catalytic wet-air oxidation (CWAO) at T=463 K in trickle-bed and batch-recycle reactors packed with either TiO2 extrudates or Ru(3 wt%)/TiO2 catalyst. Chemical analyses (TOC removal, color, HPLC) and bioassays (48-h and 30-min acute toxicity tests using Daphnia magna and Vibrio fischeri, respectively) were used to get information about the toxicity impact of the starting effluents and of the treated solutions. Under the operating conditions, complex organic compounds are mostly oxidized into carbon dioxide and water, along with short-chain carboxylic acids. Bioassays were found as a complement to chemical analyses for ensuring the toxicological impact on the ecosystem. In spite of a large decrease of TOC, the solutions of end products were all more toxic to Daphnia magna than the starting effluents by factors ranging from 2 to 33. This observation is attributed to the synergistic effects of acetic acid and salts present in the solutions. On the other hand, toxicity reduction with respect to Vibrio fischeri was achieved: detoxification factors greater than unity were measured for end-product solutions treated in the presence of the Ru(3 wt%)/TiO2 catalyst, suggesting the absence of cumulative effect for this bacteria, or a lower sensitivity to the organic acids and salts. Bleach plant effluents treated by the CWAO process over the Ru/TiO2 catalyst were completely biodegradable.     

Ecotoxicological tests assessment of soils polluted by chromium (VI) or pentachlorophenol

Carbon mineralisation and plant germination and growth (Lactuca sativa seeds) tests have been performed in two soils of different properties, experimentally spiked with pentachlorophenol (PCP) or Cr (VI), in concentrations between 0.001 and 1000 mg kg(-1). The evaluation has been done considering the following parameters of carbon transformation test: soil cumulative basal respiration after 14 days incubation, substrate induced respiration after 12 h of glucose addition, and, in the plant germination and growth test: number of germinated seeds, root elongation and total biomass produced. The most sensitive assay found in our work has been carbon mineralisation test, from which the lowest toxic concentrations were obtained (especially substrate induced respiration test). In the plant germination and growth test, the measurement of root elongation has shown the best sensitiveness, followed by plant biomass and seed germination numbers. Regarding the contaminants, the highest toxicity, considering the minimum concentration with toxic effect, has been found in PCP (0.01 mg kg(-1)) in C mineralisation test in the granitic soil. For Cr, the minimum concentration with toxic effect has been 0.1 mg kg(-1), also in the C mineralisation test and the granitic soil. The granitic soil has shown more vulnerability to the pollutants assayed in the respiration test, whereas the calcareous soil has shown more vulnerability in the plant germination and growth test.     

A comparison of microbial bioassays for the detection of aquatic toxicants

The toxicity of 35 test chemicals was analyzed using two microbial bioassay systems. The commercially available Microtox Toxicity Analyzer System™ and the two-organism procedure of Tchan were used to determine the concentration of test chemicals resulting in a 50% reduction in response (EC₅₀). Both of the tests employed a luminescent bacterium while the procedure of Tchan also utilized an alga. Results from the two microbial tests were compared with available data obtained with fish toxicity bioassays and with each other. The Microtox™ procedure was somewhat more sensitive than the Tchan bioassay in detecting most of the test chemicals and fish bioassays were generally more sensitive than either of the microbial tests. As a notable exception, photosynthesis-inhibiting herbicides were detected at remarkably lower concentrations with the procedure of Tchan than any of the other bioassays. Potential applications for these tests are discussed.     

Comparison of several microbiological toxicity screening tests

Four short-term microbiological toxicity screening tests were compared using the following test chemicals: 3,5 dichlorophenol, cetyl trimethyl ammonium chloride, sodium lauryl sulfate, phenol, copper(II) sulfate, mercury(II) chloride and zinc(II) sulfate. These seven chemicals represent a wide range of toxicity. The methods examined were Beckman's Microtox system, the Spirillum volutans motility test, inhibition of respiratory activity of activated sludge, and inhibition of activated sludge TTC-dehydrogenase activity.The results obtained indicate that each method has its own toxicity sensitivity pattern, and among the substances tested, only mercury(II) chloride and phenol were ranked equally by the four methods as the most and the least toxic chemical, respectively. In a tentative ranking of the methods, according to sensitivity, the Microtox test came out as the most sensitive test, followed by the Spirillum test, which in turn appeared more sensitive than the two sludge tests. This ranking has meaning only in a statistical sense, however, and the variable nature of the results support the philosophy that for assessing toxicity more thoroughly, a battery of several tests is required.     

Combination of in vitro bioassays encompassing different mechanisms to determine the endocrine-disrupting effects of river water

In this study, the total toxic effects of river water samples were assessed using a series of cell culture bioassays which encompassed different mechanisms, based on specific modes of action. River water samples were collected from three tributaries of the Youngsan River in the western portion of Korea. We confirmed that Youngsan River water was polluted with a complex mixture of estrogenic and dioxin-like compounds. The total toxic effects of the downstream water samples were found to be higher than that of the upstream water samples. In the upstream water samples, total estrogenic activity was measured to be between 0.005 and 0.049 ng-EEQ/l (17beta-estradiol-equivalent concentration) and no CYP1A activity was detected. In the downstream water samples, however, total estrogenic activity was measured to be between 0.021 ng-EEQ/l and 1.918 ng-EEQ/l, and total CYP1A activity was between 0.63 and 3.55 microg-MEQ/l (3-methylcholanthrene-equivalent concentration). When assessed according to a concentration-response curve, downstream water sample extracts exerted dual actions on estrogen receptors, depending on the concentration volume of the samples. The concentration volume range proximal to the original water sample exhibited estrogenic activity, whereas antiestrogenic activity was observed at high concentration volumes (more than 5 times concentration) in the extracts. This study involved a combination of in vitro bioassays, designed to encompass different mechanisms. The bioassays used included the estrogen receptor binding affinity test, E-screen assay, aromatase assay, and EROD assay. These tests provided a great deal of useful information regarding the potency and action modes of estrogenicity and antiestrogenicity inherent in the sampled river water. Although further study is necessary to determine the relationship between toxic responses in in vitro bioassay systems and chronic toxicity in aquatic organisms, our approach is expected to be fairly accurate with regard to the detection of endocrine-disrupting effects in an aquatic environment.