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.     

Identification of organic compounds and ecotoxicological assessment of sewage treatment plants (STP) effluents

An integrated approach combining chemistry and biological methods was conducted to assess the toxicity of seven sewage treatment plant effluents. Solid phase concentration procedures were applied to facilitate the study of organic micro pollutants. A chemical analysis was performed by GC/MS. Organic fraction toxicity was determined by using bioassays such as Daphnia magna and Chlorella vulgaris tests and sub-lethal effects were also evaluated by using Salmonella typhimurium Test (mutagenicity), recombinant yeast screen (estrogenicity), and Oryzias latipes embryo-larval test. More than 49 compounds were detected in the organic fraction due to the various inputs of each effluents. The most frequently detected compounds in the effluents were bisphenol A (BPA), octylphenol (OP), 1,2-benzenedicarboxylic acid, bis(2-ethylhexyl) ester (DEHP) and 1,2-benzenedicarboxylic acid, bis(methylpropyl) ester (DBP). Biological assays showed toxicity effects on D. magna tests in all samples, whereas toxicity on C. vulgaris or S. typhimurium tests were not observed. Estrogenicity and teratogenicity were observed in several samples. The cause-effect relationship could not be established given the high chemical complexity of the effluents and the lack of information available on 70% of the detected compounds subsequent to reviewing various data bases. Nevertheless, due to the high chemical variability revealed by STP effluents, bioassay sets may provide a very useful amount of information for detecting potential toxicity risks.     

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.     

Effect of pH on toxicity of kraft pulp and paper mill effluent to salmonid fish in fresh and seawater

In freshwater bioassays with juvenile rainbow trout (Salmo gairdneri), at initial pH values from 4 to 11, kraft mill effluents were considerably less toxic at pH 9–10 than at neutrality. When pH of test solutions was controlled throughout the bioassay period, the least toxic range was 8.5–9.5. Toxicity at typical receiving-water pH values was 50–67% greater.The acute toxicity of effluent samples to yearling coho salmon (Oncorhynchus kisutch) was identical for these effluents in seawater and freshwater respectively, provided that the pH was adjusted and held at the same value, and that test fish were previously acclimated to the dilution water for several months. Thus seawater constituents other than pH did not affect the acute toxicity of pulp and paper mill effluents appreciably.     

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.     

Comparison of algal assay systems for detecting waterborne herbicides and metals

The EPA Bottle Test was compared to a short-term algal bioassay method (Oxygen Evolution Assay) for detecting waterborne herbicides and metals. Although the Bottle Test proved to be a more sensitive assay system, the responses of the two bioassays correlated well. The Oxygen Evolution Assay may be useful as a rapid preliminary screening method for determining which chemicals should undergo further testing. In both bioassays, differences were observed in the results of experiments conducted in the standard algal growth medium and in nutrient-enriched water samples, suggesting that water chemistry plays an important role in the determination of specific toxicity thresholds for algae.     

Selecting a battery of bioassays for ecotoxicological characterization of wastes

This study was conducted in France within the context of waste classification (Hazardous Waste Council Directive 91/689/EEC), and focused on "ecotoxic" property (H14). In 1998, an experimental test strategy was developed to assess ecotoxicological properties of wastes using a battery of six standardized bioassays. This combined direct and indirect approaches integrating two solid-phase tests: emergence and growth inhibition of Lactuca sativa (14 days), mortality of Eisenia fetida (14 days) and four standardized tests performed on water extracts from wastes: growth inhibition of Pseudokirchneriella subcapitata (3 days), inhibition of mobility of Daphnia magna (48 h), inhibition of reproduction of Ceriodaphnia dubia (7 days), inhibition of light emission of Vibrio fischeri (30 min). This study aimed to set up preliminary conclusions on relevancy of this experimental test strategy, based on data obtained since 1998. Results were analyzed from the combined use of Hierarchical Cluster Analysis, Principal Component Analysis and Nonlinear Mapping. These multivariate analyses clearly showed that it was possible to reduce this number of tests without changing the typology of the wastes. A battery of bioassays including one solid phase test and two tests performed on water extracts (L. sativa, V. fischeri and C. dubia) was found as an optimal solution for characterizing the toxicity of the studied wastes. This optimal battery represents a good basis for determining the H14 property.     

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.     

TOXICITY TESTING OF GROUNDWATER QUALITY

Groundwater is an important resource in the UK, with 45% of public water supplies in the Thames Water region derived from subterranean sources. In urban areas, groundwater has been affected by anthropogenic activities over a long period of time and from a multitude of sources. At present, groundwater quality is assessed using a range of chemical species to determine the extent of contamination. However, analysing a complex mixture of chemicals is time-consuming and expensive, whereas the use of an ecotoxicity test provides information on (a) the degree of pollution present in the groundwater and (b) the potential effect of that pollution. Microtox^TM, Eclox^TM and Daphnia magna microtests were used in conjunction with standard chemical protocols to assess the contamination of groundwaters from sites throughout the London Borough of Hounslow and nearby Heathrow Airport. Because of their precision, range of responses and ease of use, Daphnia magna and Microtox^TM tests are the bioassays that appear to be most effective for assessing groundwater toxicity. However, neither test is ideal because it is also essential to monitor water hardness. Eclox^TM does not appear to be suitable for use in groundwater-quality assessment in this area, because it is adversely affected by high total dissolved solids and electrical conductivity.     

Bioanalytical tools for the evaluation of organic micropollutants during sewage treatment, water recycling and drinking water generation

A bioanalytical test battery was used for monitoring organic micropollutants across an indirect potable reuse scheme testing sites across the complete water cycle from sewage to drinking water to assess the efficacy of different treatment barriers. The indirect potable reuse scheme consists of seven treatment barriers: (1) source control, (2) wastewater treatment plant, (3) microfiltration, (4) reverse osmosis, (5) advanced oxidation, (6) natural environment in a reservoir and (7) drinking water treatment plant. Bioanalytical results provide complementary information to chemical analysis on the sum of micropollutants acting together in mixtures. Six endpoints targeting the groups of chemicals with modes of toxic action of particular relevance for human and environmental health were included in the evaluation: genotoxicity, estrogenicity (endocrine disruption), neurotoxicity, phytotoxicity, dioxin-like activity and non-specific cell toxicity. The toxicity of water samples was expressed as toxic equivalent concentrations (TEQ), a measure that translates the effect of the mixtures of unknown and potentially unidentified chemicals in a water sample to the effect that a known reference compound would cause. For each bioassay a different representative reference compound was selected. In this study, the TEQ concept was applied for the first time to the umuC test indicative of genotoxicity using 4-nitroquinoline as the reference compound for direct genotoxicity and benzo[a]pyrene for genotoxicity after metabolic activation.The TEQ were observed to decrease across the seven treatment barriers in all six selected bioassays. Each bioassay showed a differentiated picture representative for a different group of chemicals and their mixture effect. The TEQ of the samples across the seven barriers were in the same order of magnitude as seen during previous individual studies in wastewater and advanced water treatment plants and reservoirs. For the first time a benchmarking was performed that allows direct comparison of different treatment technologies and covers several orders of magnitude of TEQ from highly contaminated sewage to drinking water with TEQ close or below the limit of detection. Detection limits of the bioassays were decreased in comparison to earlier studies by optimizing sample preparation and test protocols, and were comparable to or lower than the quantification limits of the routine chemical analysis, which allowed monitoring of the presence and removal of micropollutants post Barrier 2 and in drinking water. The results obtained by bioanalytical tools were reproducible, robust and consistent with previous studies assessing the effectiveness of the wastewater and advanced water treatment plants. The results of this study indicate that bioanalytical results expressed as TEQ are useful to assess removal efficiency of micropollutants throughout all treatment steps of water recycling.     

Evaluation of sample preparation techniques for algal bioassays

The chemical and biostimulatory properties of mesotrophic pond water and eutrophic feedlot runoff were examined after subsamples of each were separately subjected to standard membrane filtration and autoclaving pretreatments for bottle test bioassays. Correspondingly separate subsamples were exposed to ultraviolet light to determine its effectiveness as a possible pretreatment. All three pretreatments fulfilled the requirement of destroying or removing indigenous algae, but each technique substantially altered the water chemistry changing the bioassay results. Filtration removed phosphorus; autoclaving raised the pH value, precipitated salts, and eliminated dissolved carbon dioxide; and ultra-violet light oxidized organic and inorganic nitrogen compounds.     

Toxicity assays: a way for evaluating AOPs efficiency

The technical feasibility and performance of photocatalytic degradation of aqueous methomyl (50 mg/L) have been studied at pilot scale in two well-defined systems of special interest because natural-solar UV light can be used: heterogeneous photocatalysis with titanium dioxide and homogeneous photocatalysis by photo-Fenton. The pilot plant is made up of compound parabolic collectors specially designed for solar photocatalytic applications. Experimental conditions allowed pesticide disappearance, degree of mineralisation and toxicity achieved in the two photocatalytic systems to be compared. Total disappearance of methomyl is attained by photo-Fenton in 60 min and by TiO2 in 100 min. Hundred percent of nitrogen and sulphur are recovered as ammonium and sulphate. By contrast, complete mineralisation of total organic carbon (TOC) is not achieved even after quite a long time (more than 300 min). Three different bioassays (Vibrio fischeri, Daphnia magna and a Microalga) have been used for testing the progress of toxicity during treatment. All remained toxic down to very low-pesticide concentrations and in some bioassays were still toxic after total disappearance of the pesticide. Only if treatment is maintained throughout enough mineralisation (i.e. TOC disappearance), the toxicity is reduced to below the threshold (EC50%).     

The identification of phosphorus as a growth-limiting nutrient in lough neagh using bioassays

Algae were used in monthly bioassays of Lough Neagh water to evaluate nutrient limitation. Blue-green algae were limited by chelated iron; but once this limitation was overcome both nitrogen and phosphorus were necessary to produce a large growth response. The green alga Selenastrum showed a low requirement for chelated iron. Addition of phosphorus was necessary in all bioassays for prolonged algal growth, confirming the strategy of removing phosphorus from sewage effluents to reduce algal levels in the Lough.     

Evaluation of water column and sediment toxicity from an abandoned uranium mine using a battery of bioassays

Uranium mining activities in Cunha Baixa, Mangualde (Portugal), were extensive between 1967 and 1993, with high production of poor ore. Ore exploitation left millions of tons of tailings in the surrounding area, close to human houses. Contamination of the area (water and soil compartment) presently represents a serious hazard to humans and wildlife. The aim of this work was to evaluate the acute toxicity of water and sediments from a pond that floods a uranium mine pit, in two periods (spring and autumn). High contents of metals were found in water samples (chiefly Mn, Fe, Al, U, Sr). A battery of assays was applied to screen the acute toxicity of the different compartments using algae, crustaceans and dipterans. Results showed that the sediments were non-toxic, unlike the superficial water. Water toxicity was higher in the autumn, when the effluent was more acidic, compared to spring. In the water toxicity assays, the relative sensitivity of the test species used was Daphnia longispina>Pseudokirchneriella subcapitata>Daphnia magna. The present study is part of the chemical and ecotoxicological characterisation of the aquatic compartment performed in the Tier 1 of the Ecological Risk Assessment of the Cunha Baixa mining area.     

A new method for evaluating biological safety of environmental water with algae, daphnia and fish toxicity ranks

In this study, an innovative approach to evaluate biological safety of environmental water with toxicity ranks was proposed. Widely used species, algae (Selenustrum capricornutum), daphnia (Daphnia magna) and fish (Oryzias latipes larvae) belonging to three trophic levels in aquatic ecosystem, were selected and combined as a test set to measure the bio-toxicity of water sample. Maximum exposure concentrations for algae, daphnia and fish test were respectively designed as 10-, 50- and 50-fold of river water based on a simplification of conventional toxicity extrapolation method EU Directive EEC/93/67. A novel assessment index "safety score" of 1, 2, 3 and 4 with 1 being the safest was established for normalizing the toxicity effects. Safety score was determined according to the highest exposure concentration where adverse ecotoxicological effects could not be observed, and a triangle figure was designed to visually describe the safety scores of three toxicity tests. Finally, in order to conveniently evaluate the biological safety of environmental water, an integrated assessment index "bio-safety rank" (BSR) was established and determined according to the safety scores of the three tests, and with the index BSR, water sample could be ranked as A, B, C or D with A being the safest. It was shown that the proposed new method was effective for screening and evaluating the biological safety of river water in case studies.     

Effect of organic solvents on growth of freshwater algae

The impact of acetone, methanol, ethanol and dimethyl sulfoxide (DMSO) as carrier solvents on two species of algae was investigated. Scenedesmus quadricauda of green algae and Anabaena flos‐aquae of blue green algae were subjected to different concentrations of the studied solvents which ranged from 0.01 to 7.29 mL/L for 10 days in batch studies. The data recommended the use of acetone, ethanol and methanol than dimethyl sulfoxide in this order as a carrier solvent for chemical toxicity tests using a maximum concentration of 7.29 mL/L which represents 0.729% (volume/volume) in the algal bioassay test.     

Differential sensitivity of three cyanobacterial and five green algal species to organotins and pyrethroids pesticides

In this work, five organotins and pyrethroids pesticides were tested to examine their effects on the three cyanobacteria Anabaena flos-aquae, Microcystis flos-aquae, Mirocystis aeruginosa and on the five green algae Selenastrum capricornutun, Scenedesmus quadricauda, Scenedesmus obliqnus, Chlorella vulgaris, Chlorella pyrenoidosa through 96 h acute toxicity tests. The results indicated that: (1) the decreasing order of the average acute toxicity to cyanobacteria and green algae of five dissimilar organotins and pyrethroids pesticides was: fentin hydroxide > cyhexatin > azocyclotin > fenbutatin oxide > beta-cyfluthrin. (2) Wide variations occurred in response to the tested pesticides among the eight individual species of cyanobacteria and green algae. The sensitivity of various species of algae exposed to fenbutatin-oxide varied over one order of magnitude, exposed to cyhexatin/fentin-hydroxide/beta-cyfluthrin varied over two orders of magnitude and exposed to azocyclotin varied three orders of magnitude. (3) In contrast with the sensitivity of cyanobacteria and green algae, cyanobacteria were much less sensitive to beta-cyfluthrin than green algae. The pollutants may result in a shift of green algal and cyanobacterial group structure, especially in a shift from dominance by green algae to dominance by cyanobacteria, and may sustain cyanobcterial blooms during the special period. Thus, the decreasing order of the aquatic ecological risk was: beta-cyfluthrin > fentin hydroxide > cyhexatin > azocyclotin > fenbutatin oxide. There was a strong variance between toxicity and ecological risk, i.e. "low toxicity" does not automatically imply "low ecological risk". The toxicity of pyrethroids pesticides was lower than that of organotins pesticides, whereas the aquatic ecological risk of pyrethroids pesticides was higher than that of organotins pesticides.     

The application of batch bioassay techniques to the study of salinity toxicity to freshwater phytoplankton

The effect of salinity additions on the growth of freshwater algae was studied in the laboratory utilizing batch bioassays. These batch bioassays were used to screen variations of ten salts in single addition and the concentration effects of the salts on the productivity of standard test alga and an indigenous diatom from Lake Powell. The indigenous algal genus was found to be more tolerant to salinity addition than the standard test alga.The batch bottle bioassays were conducted following the standard algal assay procedure as closely as possible. Variations in the standard algal assay procedure included media variation with the use of the indigenous alga for test inoculum in the bioassay procedure.     

Is Cr(VI) toxicity to Daphnia magna modified by food availability or algal exudates? The hypothesis of a specific chromium/algae/exudates interaction

The long-term effects of Cr(VI) on life history traits of Daphnia magna Straus were studied in relation to different feeding conditions or to the presence of Scenedesmus acutus exudates, which had proved to reduce chromium toxicity to the alga. In high feeding conditions the concentration of 14 microg Cr(VI)/L had negative effects on survival, growth and fecundity. In low feeding conditions the effects on growth were not evident, but daphnids changed the reproductive strategy normally adopted in case of food shortage and showed a daily rate of newborn production higher than control; however, newborn were of a smaller size and less resistant to starvation. The most relevant effect of Cr was a shortening of the daphnid life span, which was independent of the feeding conditions and occurred also when growth and daily rate of neonate production were not affected. The algal exudates resulted ineffective in reducing Cr toxicity to daphnids, but caused a strong decrease in daphnid fecundity. Further experiments on S. acutus demonstrated that the detoxifying effect observed in the algae might involve a specific Cr/algae/exudates interaction, which occurs only when the algae are previously subjected to a short-time stress by chromium.