Selective removal of resin and fatty acids from mechanical pulp effluents by ozone

Fresh chemithermomechanical (CTMP) mill primary clarifier (combined) effluents were treated using a system capable of accurately applying small measured charges of ozone. It was shown that the efficiency of ozone transfer into effluent was much better than into water, and that there was no residual dissolved ozone. Small ozone charges destroyed high proportions of the total resin and fatty acids (RFAs) and juvabiones (JBs) present in the effluents. This effect diminished with increasing ozone charges. Likewise, effluent toxicity, as measured in the Microtox assay, decreased rapidly with the low ozone charges. The low charges of ozone were also shown to be effective in destroying both soluble and particulate RFAs. It was shown that there was a strong positive correlation between the RFA content and the acute toxicity of an effluent sample, and that all the acute toxicity was extractable with methyl-t-butyl ether. Both Microtox and Daphnia magna acute toxicity assays correlated with effluent RFA content, but the Microtox was somewhat more sensitive. Thus, ozone may be a useful economical treatment agent for selectively removing RFAs and toxicity from mill effluent streams.     

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.     

Toxicity assessment of common xenobiotic compounds on municipal activated sludge: comparison between respirometry and Microtox

The toxicity of four xenobiotic compounds 3,5-dichlorophenol, formaldehyde, 4-nitrophenol and dichloromethane, representative of industrial wastewater contaminants was evaluated by a simple respirometric procedure set up on the basis of OECD Method 209 and by the Microtox bioassay. Very good reproducibility was observed for both methods, the variation coefficients being in the range of 2-10% for the respirometric procedure and 6-15% for Microtox, values that can be considered very good for a biological method. Comparison of EC(50) data obtained with the two methods shows that in both cases 3,5-dichlorophenol is more toxic than other compounds investigated and dichloromethane has a very low toxicity value. Intermediate EC(50) values were found for the two other chemicals, formaldehyde and 4-nitrophenol. Moreover, the Microtox EC(50) values are generally lower (except for dichloromethane) than the respirometric ones: these differences could be explained by the fact that the Microtox method uses a pure culture of marine species and, therefore, should not necessarily be expected to behave like a community of activated sludge bacteria. In conclusion, both methods can be usefully applied for toxicity detection in wastewater treatment plants but it is advisable to take into account that Microtox is more sensitive than respirometry in estimating the acute toxicity effect on the biomass operating in the plant.     

Plasticizer metabolites in the environment

Earlier work with pure cultures had shown that the interaction of microbes with plasticizers leads to the formation of metabolites including 2-ethylhexanoic acid and 2-ethylhexanol that resist further degradation. The presence of these metabolites is now reported in a variety of environmental samples. Thus, even in a complex ecosystem, when plasticizers are degraded, the breakdown is not complete and significant amounts of 2-ethylhexanoic acid and 2-ethylhexanol are observed. These compounds have been shown to exhibit acute toxicity using Microtox, Daphnia, rainbow trout and fathead minnow toxicity assays. Since it is already well established that plasticizers are ubiquitous in the environment, it is expected that their recalcitrant metabolites will also be ubiquitous. This is of concern because, while the plasticizers do not exhibit acute toxicity, their metabolites do.     

The effects of nonylphenol on Daphnia magna

The acute and chronic toxicity of nonylphenol to the freshwater invertebrate Daphnia magna has been determined. The acute 24 and 48 h EC₅₀ values, based on immobilization, were 0.30 and 0.19 mg/l, respectively, using mean measured concentrations. In a separate study, effects of nonylphenol on survival, growth and reproduction of D. magna were measured over a period of 21 days. The no observed effect concentration (NOEC), based on mean measured concentrations, was found to be 0.024 mg/l. This is approximately one order of magnitude below the acute value and one order of magnitude higher than the highest reported environmental levels.     

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 chlorpyrifos and TCP alone and in combination to Daphnia carinata: the influence of microbial degradation in natural water

The acute toxicity of chlorpyrifos and its principal metabolite 3,5,6-trichloropyridinol (TCP) alone and in combination to a cladoceran, Daphnia carinata, was studied in both cladoceran culture medium and natural water collected from a local suburban stream. TCP was found to be more toxic than its parent chemical chlorpyrifos to Daphnia survival in cladoceran culture medium. However, TCP in natural water was not toxic to D. carinata up to 2 microgL(-1). The LC(50) values for chlorpyrifos, TCP and chlorpyrifos+TCP were 0.24, 0.20 and 0.08 microgL(-1), respectively, in cladoceran culture medium. Although the parent chemicals and their degradation products co-exist in natural waters, the existing guidelines for water quality are based on individual chemicals. The results of this investigation suggest that chlorpyrifos and TCP can interact synergistically, additively or antagonistically, resulting in an increase or decrease in the overall toxicity of the mixture compared to individual compounds. The indigenous microorganisms in natural water could play a significant role in degradation of these compounds thereby influencing their toxicity in receiving waters. This study clearly suggests that the joint action of pesticides and their degradation products should be considered in the development of water quality guidelines. To our knowledge, this is the first study on the interactive effect of chlorpyrifos and TCP to a cladoceran and suggests that these two compounds are non-toxic when present together at concentrations up to 0.12 microgL(-1). However, these compounds together act additively at and above 0.5 microgL(-1) to fresh water invertebrates and therefore pollution with these compounds may adversely affect natural ecosystems.     

Assessing the influence of methanol-containing additive on biological characteristics of diesel exhaust emissions using microtox and mutatox assays

Here we investigate the effect of the methanol-containing additive (MCA) on the biological characteristics of diesel exhaust emissions. Microtox and Mutatox assays, respectively, were used to evaluate the acute toxicity and genotoxicity of crude extracts from diesel engine exhaust. The engine was tested on a series of diesel fuels blended with five additive levels (0, 5, 8, 10 and 15% of MCA by volume). Emission tests were performed over the hot start portion of the transient Heavy-Duty-Federal Test Procedure (HD-FTP) and two selected steady-state modes. Microtox results show that MCA additive moderately lowers the toxicity levels of particle-associated (SOF) samples, but generally increase the vapor-phase (XOC) associated toxicity. A strong correlation was found between XOC-associated toxicity and total hydrocarbon (THC) concentrations, while only a slight link was found between SOF-associated toxicity and particulate matter (PM) concentrations. For Mutatox test results, when either 5 or 8% MCA used, XOC and SOF-associated genotoxicity in both steady-state and hot-start transient cycle tests were relatively lower compared to those of the base diesel. The genotoxic potential of XOC samples was significantly increased after treatment with an exogenous metabolic activation system (S9). On the contrary, the genotoxic potential of SOF samples without S9 metabolic activation was generally higher than those with S9. It is noteworthy that the total particle-associated (SOF) PAHs emissions showed trends quite similar to that of the genotoxic potential. As expected, the total particle-associated (SOF) PAHs correlated moderately with direct mutagenicity, and fairly well with indirect mutagenicity. Finally, the genotoxicity data did not parallel the Microtox results in this study, indicating that potentially long-term genotoxic agents may not be revealed by short-term toxicity assays.     

Toxicity of the 13 priority pollutant metals to Vibrio fisheri in the Microtox chronic toxicity test

The Microtox Acute Toxicity Test has been successfully used to measure the toxicity of metals and other pollutants at high concentrations (ppm) in selected environmental samples. However, metals and other toxicants are often found in much lower concentrations (ppb) in many municipal wastewaters and receiving waters. In order to assess the toxicity of these pollutants in these samples, a more sensitive toxicity assay is needed. The Microtox chronic toxicity test has been developed to measure the sublethal effect of toxicants over multiple generations of the test species, Vibrio fisheri. In this study, the toxicity of the 13 priority pollutant metals [i.e. As, Se, Cd, Cr (III and VI), Cu, Pb, Sb, Ag, Tl, Zn, Be, Hg and Ni] to V. fisheri was evaluated using the Microtox chronic toxicity test. In this test, the inhibitory concentration (IC), lowest observable effect concentration (LOEC), and no observable effect concentration (NOEC) were obtained after 22-h of incubation at 27+/-1 degrees C, by comparing the light output of the control to that of the test sample. Among the 13 priority pollutant metals, beryllium (Be) was found to be the most toxic in the test (LOEC=0.742-1.49 microg/l) while thallium (Tl) was the least toxic (LOEC=3840-15300 microg/l). The LOECs for copper (as Cu) and lead (Pb) in reagent (ASTM Type I) water were 6.78-13.6 microg/l and 626-1251 microg/l, respectively. The toxicity of copper sulfate (as Cu) in reagent water was shown and significantly reduced with the addition of natural organic matter (fulvic acid) or EDTA to the sample. The LOEC values for the 13 priority pollutant metals in this test were comparable to or lower than those reported for commonly used aquatic toxicity tests, such as the Ceriodaphnia dubia assay.     

Contribution for tier 1 of the ecological risk assessment of Cunha Baixa uranium mine (Central Portugal): I soil chemical characterization

Within the tier 1 of a site specific risk assessment, the pseudo-total concentrations (extracted with aqua regia) and the potential mobile fractions of metals were determined to perform a preliminary evaluation of risks posed by contaminated soils from an abandoned uranium mine (Mangualde, Central Portugal). Considering the mobile fractions of metals, extracted with artificial rain water, aluminium and uranium were the most concerning elements, since their concentrations were above soil quality criteria values (SQGVs) established for both elements. However, according to the evaluation based on potential mobile fractions of elements, rather than on pseudo-total metal concentrations the risks were limited to sites within the exploitation area, where contamination derives mainly from past in-situ leaching activities of pore ore as well as from the deposition of sludge from the effluent pond. The exclusion of other sites under evaluation, from the risk assessment process, requires additional data provided by soil screening ecotoxicological assays.     

Evaluation of wastewater toxicity: comparative study between Microtox and activated sludge oxygen uptake inhibition

Microtox is a frequent toxicity tool for the screening of wastewaters discharged into wastewater treatment plants. There is currently an increasing controversy between this test and others using activated sludge. A Microtox and electrolytic respirometry comparative study for toxicity determination has been performed. Seven organic and five inorganic toxic compounds have been assessed for comparing both methods. Microtox proved to have a higher sensitivity to toxicants but was less representative of effects on activated sludge compared to respirometry. For instance, assays accomplished with LAS, a biodegradable reference surfactant, showed a toxic effect by Microtox but good biodegradability and no toxicity in respirometry. This could be explained by the different nature of the biological material used, as Microtox utilises the seawater Vibrio fischeri, whereas respirometry uses the bacterial consortium in activated sludge. For the evaluation of the potential toxicity of a compound on a WWTP, the preferred biological material be used should be activated sludge itself. Results obtained with any other biological material would be just an approach to reality.     

Comparative temporal ecotoxicological study in a river basin influenced by petrochemical industries

An approach was developed to evaluate ecotoxicological effects in river basins impacted by anthropic industrial discharges. Genotoxicity was the first level of evaluation of the river water, but when cytotoxicity was associated with genotoxic effects, our research group added chronic assays allowing the assessment of the ontogenic cycle in environmental diagnosis. The genotoxicity of river water sampled during two periods was compared using the microscreen phage-induction and Salmonella/microsome assays. The study assessed the ability of these assays to diagnose environmental quality in an area where petrochemical impacts occur. The study was performed at six sampling sites using the Salmonella/microsome assay for 10 sampling periods and the microscreen phage-induction assay for 15 samplings. The percentage of mutagenic activity was higher at the sites sampled in front of the petrochemical complex in both periods, but there were more significant mutagenic responses in the first assessment. However, comparing the percentages of samples with mutagenic and cytotoxic activity observed during the two periods it becomes clear that there are more cytotoxic samples during the second period throughout the area studied. The genotoxic activity analyzed by the microscreen phage-induction assay was constant in the second period. Chronic toxicity studies with the microcrustacean Daphnia magna confirm the toxic effects observed. The mortality of individuals was higher at the site most influenced by the petrochemical complex, followed by the station located upstream from this area, while the formation of ephippial eggs was uniform at all stations. The rise in the toxic potential of the region detected by cytotoxic and chronic toxicity may interfere and probably impair the use of the Salmonella/microsome assay to determine the potential of the area, showing the importance of assay association to evaluate potentially contaminated areas.     

Effects of dissolved humic materials on acute toxicity of some organic chemicals to aquatic organisms

The influence of dissolved humic materials (DHM) on the acute toxicity of diazinon, tetrabromobisphenol-A (TBP), 4-chloroanilin (4-CA) and pentachlorophenol (PCP) was evaluated using the zebrafish (Brachydanio rerio Hamilton-Buchanan) and waterflea (Daphnia magna Straus). The 96-h LC₅₀ (zebrafish) and 48-h EC₅₀ values for four chemicals were determined in the presence of 0, 0.5, 5.0 and 50 TOC mg/l. The effects of DHM on the toxicity of four test chemicals to B. rerio were not observed in all of the DHM treatments. In the daphnid toxicity test, DHM significantly reduced the toxicity of diazinon and 4-CA, while no reduction of the toxicity of TBP and PCP was observed. These results indicate that the interaction between DHM and chemicals can alter the toxicity of some chemicals to D. magna.     

Toxicity to Daphnia of the end products of wet oxidation of phenol and substituted phenols

The process of wet oxidation breaks down organic substances in aqueous solution at elevated temperatures and pressures. Experimental wet oxidations were carried out on pure solutions of phenol, 2-chlorophenol and 4-nitrophenol. After 1-h wet oxidation, final concentrations of these compounds averaged 3% of their concentrations in the starting solutions. The toxicities of the starting compounds and the residual toxicity of the end-product solutions were measured with 48-h acute toxicity tests using Daphnia magna. The solutions of end products were all less toxic than the starting solutions by factors ranging from 10 to 120. However, the end-product solutions were somewhat more toxic than would be predicted from the known concentration of initial compound remaining in the solution of end products.     

Preliminary toxicity assessment of water after treatment with UV-irradiation and UVC/H2O2

Photooxidation (UV radiation) and enhanced photooxidation (UVC/H2O2) are water treatment technologies which remove aquatic natural organic matter (NOM) by photodegradation, producing lower molecular weight components and CO2. Since these technologies are being investigated for the treatment of drinking water, knowledge of the potential toxicity of the photooxidation by-products is vital. The potential toxicity of UVA-, UVB-, UVC-irradiated, and UVC/H2O2-treated aquatic NOM in two spot samples from two Australian reservoirs was analysed in two spot samples using Vibriofischeri in the Microtox test, African green monkey kidney cells (AS/NZS 4020:1999), and Daphnia carinata in an acute immobilisation test. Toxicity was not apparent for both the Microtox procedure and cytotoxicity analyses for the UVC-irradiated and UVC/H2O2-treated NOM samples, while UVA- and UVB-irradiated water samples were non-toxic to D. carinata. In contrast, acute toxicity was observed for UVC- and UVC/H2O2-treated water samples. The observed toxicity was attributed to photooxidative degradation of NOM-metal binding sites, which resulted in the release of bioavailable copper ions, as evidenced by higher concentrations of free copper ions in photooxidised water. As the total copper concentrations of the two raw water samples were well below the Australian Water Quality Guidelines for metals in domestic supplies, the release of copper from photooxidised NOM is unlikely to cause health concerns in these samples.     

Toxicological and chemical characterization of organic micropollutants in river po waters (Italy)

River Po waters were collected at the end of the drainage basin, extracted by means of XAD-2 resins and tested for toxicity on aquatic organisms (Daphnia magna and Selenastrum capricornutum) and for mutagenicity with a modified Ames test. The extracts were analyzed by HRGC/FID and selective detectors for the determination of the most common classes of toxic compounds. The relationships between toxicological responses and analytical results are discussed.     

Aquatic microbial activity and macrofaunal profiles of an Oklahoma stream

A toxicity assessment was conducted on Skeleton Creek, near Enid, Okla. This creek received effluents from an oil refinery, municipal waste treatment facility, and a fertilizer processing plant. Microbial activity assays, which focused on sediments, included: dehydrogenase, alkaline phosphatase, glucosidase, amylase and protease activity. Microbial electron transport system activity was also measured in overlying waters and sediments. Results were compared to a study conducted at the same time by the U.S. Environmental Protection Agency (EPA) which measured in-stream fish, plankton and benthic communities. In addition, EPA conducted 7-day Ceriodaphnia reproductive toxicity tests and 7-day larval fathead minnow growth tests. Effects occurred below the 3 discharges and similar response patterns were noted between surrogate microbial assays and stream communities. These results highlight the importance and feasibility of multi-tiered test schemes in aquatic toxicity assessments.     

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.     

Risk-based ecological soil quality criteria for the characterization of contaminated soils. Combination of chemical and biological tools

This paper describes the development of soil quality criteria for the characterization of soils focused on the potential risk to the ecosystem. The approach combines both Generic Soil Quality standards (GSQs) for individual chemicals and direct ecotoxicity assays on soil samples taken from the site. Criteria establish three main risk levels with their corresponding trigger values. The trigger values to determine high risk or "polluted" soils are exclusively based on direct toxicity assessments. The trigger values for the other categories are established by a combination of the application of GSQs and the results of bioassays. Low-risk is assumed when no toxicity is observed and GSQs based on precautionary ecotoxicity thresholds are not exceeded; high-risk must be considered if acute toxicity above the proposed trigger value is observed in soil or leachate samples. In between these levels, the risk cannot be elucidated and a site-specific assessment is required. The GSQs take into account the current or future land use, thus defining three categories: industrial soils, urban/residential soils and natural/agricultural/forest soils, each of them with different ecological requirements. The GSQ values are established following an inverse risk assessment methodology, integrating ecotoxicity and exposure models and setting the soil levels associated to pre-established criteria for the assumption of low risk. The proposed methodology covers all relevant ecological receptors and processes, soil organisms, potential contamination of ground and surface waters, and exposure of terrestrial vertebrates due to bioaccumulation and biomagnification. Exposure routes and protection criteria are defined in each protection goal. The relevance of each receptor and route is established according to the land use.     

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.