Daphnia Magna Toxicity Research Articles

Toxicity and genotoxicity of hospital laundry wastewaters treated with photocatalytic ozonation

The aim of the present study was to assess the efficiency of advanced oxidative processes based on photocatalytic ozonation (O3, UV, UV/O3, UV/O3/Fe2+ 50mgL−1 and 150mgL−1) in the treatment of hospital laundry wastewaters. The analysis of the investigated wastewater revealed high chemical oxygen demand (COD — 3343.8mgL−1), biochemical oxygen demand (BOD5 — 1906.4mgL−1), total Kjeldahl nitrogen (TKN — 79.8mgL−1) and Daphnia magna toxicity (EC50=1.73). Genotoxic effects were also detected for Allium cepa. Reductions of some parameters occurred after photocatalytic ozonation. The UV/O3/Fe2+ 150mgL−1 method was more efficient in reducing COD (59.1%), BOD5 (50.3%) and TKN (86.8%). There was significant reduction (p<0.05) in D. magna toxicity, O3 (EC50=47.3%), UV (EC50=50.6%) and UV/O3/Fe2+ 150mgL−1 (EC50=45.4%) processes. Normalization of the mitotic index and reduction of micronucleated cells were observed in A. cepa after the treatments. Results demonstrate that these methods were efficient in the degradation of hospital laundry wastewaters, representing a thriving alternative for the removal of pollutants that cause toxicity and genotoxicity.

Organic carbon content effects on bioavailability of pyrethroid insecticides and validation of Solid Phase Extraction with Poly (2,6-diphenyl-p-phenylene oxide) Polymer by Daphnia magna toxicity tests

Solid phase extraction with Poly (2,6-diphenyl-p-phenylene oxide) Polymer (Tenax) was used for determining the bioavailability of eleven pyrethroids in field collected sediments with different organic carbon content (OC). The bioavailable fraction of pyrethroids decreased with increasing OC in sediments; the percentages of desorption ranged from 10 to 20% for sediment with higher OC content (5.8%) and 15–40% for that with lower OC (2%). Generally pyrethroids showed low bioavailability and cyfluthrin resulted to be the most bioavailable among the studied pyrethroids. Acute toxicity tests with Daphnia magna were carried out on sediment spiked with three selected pyrethroids (λ-cyhalothrin, cypermethrin and deltamethrin) and served to validate the efficiency of Tenax as a method for assessing the bioavailability of pyrethroids. Toxicity test demonstrated that Tenax was able to remove the toxic bio-available fraction of pyrethroids in sediment. Extracts from Tenax beads after the desorption experiments and spiked sediment before desorption had an equivalent toxicity (LC50) to D. magna neonates at 48 and 72h of exposure. These results indicate that Tenax beds can be used to predict bio-available and toxic fractions of pyrethroids sorbed to sediments to aquatic organisms like D. magna.

Impact of biodegradation on the potential bioaccumulation and toxicity of refinery effluents

Whole effluent assessments (WEA) are being investigated as potential tools for controlling aqueous industrial discharges and minimizing environmental impact. The present study investigated how toxicity and the presence of potentially bioaccumulative substances altered when refinery effluents were subjected to biodegradation tests. Three petrochemical effluents were assessed, two freshwater and one saline, and subjected to two different types of biodegradation tests, resembling either a ready style (dissolved organic carbon (DOC)-die away) or an inherent style (Zahn-Wellens) test and the toxicity and potential to bioaccumulate parameters were re-analysed during and after biodegradation. A high proportion of the potentially bioaccumulative substances (PBS) in these effluents was easily biodegradable. Biodegradation not only lowered the PBS concentration but also toxicity. Appropriate controls are required however, as some increases in toxicity were observed after 4 h. In the present study, six other petrochemical effluents were also assessed for their PBS content and toxicity to increase the understanding of the relationship between PBS and toxicity. The results showed that the PBS concentrations in these samples were lower than the estimated benchmarks of acute toxicity for algae, fish and crustacean, although two samples were above the critical PBS values for chronic narcotic toxicity for Daphnia magna, which support the assumption that narcotic effects are mainly responsible for the observed toxicity in refinery effluents. It can be concluded that for facilities processing petroleum products that the measurement of PBS is a suitable surrogate for toxicity tests at the screening stage. Finally, the combination of persistency, bioaccumulation, and toxicity tests was shown to have additional value compared to an approach using only toxicity tests.

Biological Surface Coating and Molting Inhibition as Mechanisms of TiO2 Nanoparticle Toxicity in Daphnia magna

The production and use of nanoparticles (NP) has steadily increased within the last decade; however, knowledge about risks of NP to human health and ecosystems is still scarce. Common knowledge concerning NP effects on freshwater organisms is largely limited to standard short-term (≤48 h) toxicity tests, which lack both NP fate characterization and an understanding of the mechanisms underlying toxicity. Employing slightly longer exposure times (72 to 96 h), we found that suspensions of nanosized (∼100 nm initial mean diameter) titanium dioxide (nTiO2) led to toxicity in Daphnia magna at nominal concentrations of 3.8 (72-h EC50) and 0.73 mg/L (96-h EC50). However, nTiO2 disappeared quickly from the ISO-medium water phase, resulting in toxicity levels as low as 0.24 mg/L (96-h EC50) based on measured concentrations. Moreover, we showed that nTiO2 (∼100 nm) is significantly more toxic than non-nanosized TiO2 (∼200 nm) prepared from the same stock suspension. Most importantly, we hypothesized a mechanistic chain of events for nTiO2 toxicity in D. magna that involves the coating of the organism surface with nTiO2 combined with a molting disruption. Neonate D. magna (≤6 h) exposed to 2 mg/L nTiO2 exhibited a “biological surface coating” that disappeared within 36 h, during which the first molting was successfully managed by 100% of the exposed organisms. Continued exposure up to 96 h led to a renewed formation of the surface coating and significantly reduced the molting rate to 10%, resulting in 90% mortality. Because coating of aquatic organisms by manmade NP might be ubiquitous in nature, this form of physical NP toxicity might result in widespread negative impacts on environmental health.

Aggregation and dispersion of silver nanoparticles in exposure media for aquatic toxicity tests

Silver nanoparticles (AgNPs) are currently being very widely used in industry, mainly because of their anti-bacterial properties, with applications in many areas. Once released into the environment, the mobility, bioavailability, and toxicity of AgNPs in any ecosystem are dominated by colloidal stability. There have been studies on the stability or the aggregation of various nanoparticles (NPs) under a range of environmental conditions, but there is little information on fully characterised AgNPs in media used in (eco)toxicity studies. In this study, monodisperse 7, 10 and 20 nm citrate-stabilised AgNPs were synthesised, characterised and then fractionated and sized by flow field-flow fractionation (FFF) and measured with dynamic light scattering (DLS) in different dilutions of the media recommended by OECD for Daphnia magna (water flea) toxicity testing. Stability of NPs was assessed over 24 h, and less so over 21 days, similar time periods to the OECD acute and chronic toxicity tests for D. magna. All particles aggregated quickly in the media with high ionic strength (media1), resulting in a loss of colour from the solution. The size of particles could be measured by DLS in most cases after 24 h, although a fractogram by FFF could not be obtained due to aggregation and polydispersity of the sample. After diluting the media by a factor of 2, 5 or 10, aggregation was reduced, although the smallest NPs were unstable under all media conditions. Media diluted up to 10-fold in the absence of AgNPs did not induce any loss of mobility or fecundity in D. magna. These results confirm that standard OECD media causes aggregation of AgNPs, which result in changes in organism exposure levels and the nature of the exposed particles compared to exposure to fully dispersed particles. Setting aside questions of dose metrics, significant and substantial reduction in concentration over exposure period suggests that literature data are in the main improperly interpreted and nanoparticles are likely to have far greater biological effects than suggested thus far by poorly controlled exposures. We recommend that the standard OECD media is diluted by a factor of ca. 10 for use with these NPs and this test media, which reduces AgNP aggregation without affecting the viability of the text organism.

Evaluation of buffer zone effectiveness in mitigating the risks associated with agricultural runoff in Prince Edward Island

To minimize the risk posed by runoff from row crops, Prince Edward Island introduced buffer legislation in 2000. The legislation mandates 10-m and 20-m buffers, respectively, for moderate sloped (i.e. < 5%) and steep sloped (i.e. > 5%) agricultural fields that border streams. Since 2001, Environment Canada has been evaluating the effectiveness of various buffer widths on operational farms in reducing toxicity and contaminant concentrations in runoff. Sample collectors, placed in 44 fields at the field edge (0 m), 10 m and at distances out to 30 m, collected overland flow following rainfall-induced runoff events. Samples were collected within 24 hours of an event and analysed for seven pesticides (endosulfan, chlorothalonil, carbofuran, linuron, metribuzin, metalaxyl, mancozeb), water quality parameters and Daphnia magna toxicity. The 10-m buffer required for moderate sloped fields was effective at reducing contaminant concentrations but not always to less than lethal concentrations to Daphnia magna. Limited data beyond 10 m for fields of both slope types precluded making recommendations on a suitable buffer width for shallow sloped fields and evaluating the effectiveness of 20-m buffers for steep sloped fields. When paired data were combined and statistically tested for all fields, the studied pesticides underwent a 52-98% and 68-100% reduction in aqueous and particulate concentrations within 10 m and 30 m, respectively. In addition, by 10 m, soluble phosphorus, nitrate-nitrogen and total suspended solids were reduced by 34%, 38% and 64%, respectively. Results suggest buffer zones on operational farms are capable of achieving contaminant reductions comparable to those reported for controlled experiments. Inconsistent siting of sample collectors beyond 10 m limited the evaluation of the effects of field slope and buffer width on buffer effectiveness on working farms. Future studies on buffer efficiency on operational farms should focus on building the data set beyond 10 m and evaluating load reductions.

Sensitivity of bacterial vs. acute Daphnia magna toxicity tests to metals

Abstract The objectives of this study were to evaluate the sensitivity of two bacterial tests commonly used in metal toxicity screening — the Vibrio fischeri bioluminescence inhibition test and the Pseudomonas putida growth inhibition test — in comparison to the standard acute Daphnia magna test, and to estimate applicability of the selected methods to the toxicity testing of environmental samples. The D. magna acute test proved to be more sensitive to cadmium (Cd), zinc (Zn) and manganese (Mn) than the two bacterial assays, whereas P. putida seems to be the most sensitive species to lead (Pb). Manganese appears to be slightly toxic to D. magna and non-toxic to the two selected bacteria. This leads to the conclusion that even in regions with high background concentrations, manganese would not act as a confounding factor. Low sensitivity of V. fischeri to heavy metals questions its applicability as the first screening method in assessing various environmental samples. Therefore, it is not advisable to replace D. magna with bacterial species for metal screening tests. P. putida, V. fischeri and/or other bacterial tests should rather be applied in a complex battery of ecotoxicological tests, as their tolerance to heavy metals can unravel other potentially present toxic substances and mixtures, undetectable by metal-sensitive species.

Influence of pH, Hardness, Dissolved Organic Carbon Concentration, and Dissolved Organic Matter Source on the Acute Toxicity of Copper to Daphnia Magna in Soft Waters: Implications for the Biotic Ligand Model

The influence of pH, dissolved organic carbon (DOC) concentration, water hardness, and dissolved organic matter (DOM) source on the acute toxicity of copper were investigated with standardized 48-h Daphnia magna toxicity tests. Toxicity tests were conducted according to a four-factor complete factorial design. Nominal factor levels were as follows: pH 6 and 8; DOC, 2.5 and 10 mg/L; hardness, 10, 20, and 40 mg/L as CaCO3; and two DOM sources (collected from the Black River and Edisto River, SC, USA). The experimental design resulted in 24 different factor level combinations. Results indicated that all factors had significant effects on copper toxicity. Furthermore, a strong interactive effect of DOC concentration and pH was detected. Because the biotic ligand model (BLM) has become a widely used tool for predicting toxicity and interpreting toxicity test results, its performance with these data was evaluated. Seventy percent of BLM predictions were within twofold of the observed median lethal concentrations. However, BLM parameters could be adjusted to improve model performance with this data set. This analysis suggested that in soft waters, the CuOH+ complex binds more strongly with the biotic ligand and that the competitive effect of hardness cations should be increased. The results of the present study may have implications for application of the BLM to some types of surface waters. Furthermore, a comprehensive analysis of BLM performance with all available data should be performed, and necessary updates to model parameters should be made to produce the most robust and widely applicable model.

Biodegradation and detoxication of phenol by using free and immobilized cells of Acinetobacter sp. XA05 and Sphingomonas sp. FG03

Strain XA05 and FG03 with high biodegradation activity of phenol were isolated from the activated sludge and phenol-contaminated soils in Northwest of China, respectively. DNA sequencing and homologous analysis of 16s rRNA gene identified that XA05 belonged to an Acinetobacter sp. and FG03 was closely related to the Sphingomonas sp. Cells of strain XA05 and FG03 were mixed at the ratio of 1:1, and polyvinyl alcohol (PVA) was used as a gel matrix to immobilize mixed cells by repeated freezing and thawing. Biodegradation was evaluated by determining phenol. Detoxication was evaluated by using Daphnia magna toxicity tests. The removal effciency of phenol and factors affecting phenol degradation were investigated, the stability of the immobilized cells was also reported. Experimental values indicated that both free cells and immobilized cells showed high phenol degradation effciencies, higher than 95% within 35 h with an initial concentration of 800 mg/L phenol, and the immobilized cells showed better performance than that of the suspended-culture cells. These results indicate that immobilized Acinetobncter sp. XA05 and Sphingomonas sp. FG03 possesses a good application potential in the treatment of phenol-containing wastewater.

Generation of endocrine disruptor compounds during ozone treatment of tannery wastewater confirmed by biological effect analysis and substance specific analysis

Ozone (O3) with its high oxidation potential was used to degrade or eliminate pollutants contained in tannery wastewater when applying different pHs and quantities of O3. Our objective was a chemical degradation by O3 to achieve an enhancement of biodegradability, with a parallel decrease in toxicity. Conventional analyses and bioassays beside substance specific analyses were performed to clear-up the behaviour of wastewater content from tanning process. The results demonstrate that the dominant organic pollutants were chemically degraded by oxidation as the chemical and biochemical oxygen demand (COD and BOD) prove, while changes in carbon content monitored by total or dissolved organic carbon content (TOC or DOC) were only marginal. Vibrio fischeri and Daphnia magna toxicity testing performed in parallel proved a decrease in toxicity after O3-treatment, while the estrogenic activity determined by enzyme-linked receptor assay (ELRA), however, proved an increase of endocrine disruptor compounds (EDC). Results could be explained by substance-specific analyses using gas chromatography (GC-MS) and liquid chromatography/mass spectrometry (LC-MS). From GC-MS analysis the elimination of non-polar compounds could be recognized, whereas the oxidative conversion led to an increase of EDC compounds, which qualitatively could be identified by LC-MS as nonylphenol ethoxylate (NPEO) degradation products: short chain NPEOs, nonylphenol carboxylates (NPECs) and nonylphenol (NP).

Classificatory and quantitative models of the relationship between the structures of chemical compounds and their toxicity for Daphnia magna

A structural similarity method was used to construct stable classificatory models for discriminating compounds with mechanisms of general toxicity to Daphnia magna compared with all other compounds with different actions, compounds with the polar narcosis mechanism compared with all others, compounds inhibiting cholinesterase compared with compounds with other mechanisms, and compounds with general toxic actions compared with substances with the polar narcosis mechanism. Simultaneous use of the classificatory and regression models allowed clear identification of substances acting via the general toxicity mechanism, with quantitative assessments of LC50 values and analysis of contributions of steric interactions and hydrogen bonds to toxicity. Our own experimental data on the toxicities (LC50) of 119 chemical compounds with different types of toxic actions to Daphnia magna are presented.

Does sulfide or water hardness protect against chronic silver toxicity in Daphnia magna? A critical assessment of the acute-to-chronic toxicity ratio for silver

The protective effects of sulfide and water hardness against acute and chronic silver toxicity in Daphnia magna were assessed in the presence of food. Results showed that both sulfide and water hardness protected against lethal acute and chronic silver toxicity in terms of mortality. However, only sulfide showed a protective effect against the sub-lethal chronic silver effects on growth and reproduction. These findings suggest that both reactive sulfide and water hardness must be taken into account in the development of a chronic version of the Biotic Ligand Model (BLM) for waterborne silver. Furthermore, acute-to-chronic ratio values for silver toxicity showed that only small increases in toxicity are seen over the chronic exposure relative to the acute toxicity. Mortality is the most sensitive endpoint in moderately hard water and in the presence of sulfide. Reproduction, measured as the number of neonates produced per adult per reproduction day, is the most sensitive one in hard water in the absence of sulfide.

Degradation and detoxification of cresols in synthetic and industrial wastewater by an indigenous strain of Pseudomonas putida in aerobic reactors

We studied the degradation of mixtures of o-cresol, m-cresol, and p-cresol, by Pseudomonas putida isolated from natural sources, and the application of this degradation to the depuration and detoxification of synthetic and industrial wastewater. Biodegradation assays were performed in batch and continuous-flow fixed-bed aerobic reactors. Biodegradation was evaluated by cresol determination using micellar electrokinetic capillary chromatography, UV spectrophotometry, and chemical oxygen demand (COD). Mineralization of cresols was assessed by gas chromatography performed both at the end of the batch process and in the continuous flow reactor effluent. Microbial growth was measured by the plate count method. Scanning electronic microscopy was employed to observe bacterial cells adsorbed on polyvinyl chloride cylinders in the reactor. Detoxification was evaluated by Vibrio fischeri, Pseudokirchneriella subcapitata, and Daphnia magna toxicity tests. Results obtained show that under batch conditions the strain grew exponentially with 100, 200, and 300 mg/L of each of the isomers in synthetic minimal medium within 48 h; in industrial wastewater with 540 mg/L of cresols similar results were obtained. Removal of cresols and COD was higher than 99.9% and 95.0%, respectively. When assays were performed in continuous flow reactor in synthetic wastewater under operating conditions a removal of total cresols and COD of 99.9% and 96.4%, respectively, was achieved. Results of capillary electrophoresis may suggest a concurrent isomers utilization and simultaneous growth on the substrates. Toxicity was neither detected at the end of the batch process nor in the continuous flow reactor effluent.

Effects of waterborne uranium on survival, growth, reproduction and physiological processes of the freshwater cladoceran Daphnia magna

Acute uranium toxicity (48 h immobilisation test) for Daphnia magna was determined in two different exposure media, differing in pH and alkalinity. LC 50 varied strongly between media, from 390 ± 40 μg L −1 U at pH 7 to 7.8 ± 3.2 mg L −1 U at pH 8. According to the free ion activity model uranium toxicity varies as a function of free uranyl concentration. This assumption was examined by calculating uranium speciation in our water conditions and in those reported in the literature. Predicted changes in free uranyl concentration could not solely explain observed differences in toxicity, which might be due to a competition or a non-competitive inhibition of H + for uranium transport and/or the involvement of other bioavailable chemical species of uranium. Chronic effects of uranium at pH 7 on mortality, ingestion and respiration, fecundity and dry mass of females, eggs and neonates were investigated during 21-day exposure experiments. A mortality of 10% was observed at 100 μg L −1 U and EC 10 for reproduction was 14 ± 7 μg L −1 U. Scope for growth was affected through a reduction in feeding activity and an increase in oxygen consumption at 25 μg L −1 U after 7 days of exposure. This had strong consequences for somatic growth and reproduction, which decreased, respectively, by 50% and 65% at 50 μg L −1 U after 7 days and at 25 μg L −1 U after 21 days. Uranium bioaccumulation was quantified and associated internal alpha dose rates from 2.1 to 13 μGy h −1 were estimated. Compared to the toxicity of other alpha-emitting radionuclides and stable trace metals, our results confirmed the general assumption that uranium chemical toxicity predominates over its radiotoxicity.

Recovery of N and P from human urine by freezing, struvite precipitation and adsorption to zeolite and active carbon

The majority of the nutrients in domestic waste originate from human urine. This study deals with methods for recovery of N and P from urine. Results from a freezing-thawing method (FTM) together with struvite recovery and nitrogen adsorption on zeolite and active carbon (AC) are presented. Various amounts of MgO, zeolite and AC were added to samples of 100 ml urine. After 3 days the supernatants were analysed for pH, total-N, total-P and acute toxicity for Daphnia magna. One set of samples was frozen and then thawed and the supernatants collected were tested as before. The FTM method concentrated 60% of the nutrients in 40% of the initial volume and significantly improved the N reduction and D. magna survival. The P recovery was 95–100%, mainly as struvite. No significant effect of AC was found. Zeolite improved the P recovery and in some combinations of MgO also the N recovery.

The applicability of acetylcholinesterase and glutathione S-transferase in Daphnia magna toxicity test

The most commonly used toxicity test worldwide is the acute Daphnia magna test. The relevance of acetylcholinesterase (AChE) and glutathione S-transferase (GST) activity in D. magna exposed to chromium, cadmium, and diazinon was evaluated in connection with this standard test. We found no link between enzyme activities and immobility. Concentrations of Cr 6+ up to 280 μg/L had no effect on AChE and GST activities, while 20% immobility was observed. At concentrations of 20–25 μg/L of Cd 2+ AChE activity was increased by about 50%. The effect of diazinon on both enzymes was insignificant up to concentrations that caused 27% immobility. Consequently, while the use of AChE and GST activities is recommended when the mode of action of chemicals is studied, the value of these biomarkers in routine acute toxicity tests is limited because the relationship between enzyme activities and immobility of D. magna exposed to different chemicals is unclear.

Effect of Perozonation on Biodegradability and Toxicity of a Penicillin Formulation Effluent

The pretreatment of synthetic penicillin formulation effluent containing Procain Penicillin G (PPG) with the O3/H2O2 process (applied ozone dose = 1440 mg h−1 treatment time = 60 minutes; pH 7; H2O2 = 10 mM) was investigated. The effect of chemical pretreatment was assessed on the basis of acute toxicity and biodegradability with activated sludge using water flea Daphnia magna toxicity and activated sludge inhibition tests. Biological treatability studies were performed with a mixture of untreated or pretreated PPG effluent (25% on volume basis) and synthetic domestic wastewater simulating readily biodegradable organic substrate to simulate the characteristics of domestic wastewater (75% on volume basis). Pretreatment of PPG effluent the O3/H2O2 process resulted in more than 70% chemical oxygen demand (COD) removal and a 50% decrease in the acute toxicity towards Daphnia magna. On the other hand, biodegradation of untreated PPG effluent needed prolonged acclimation periods to obtain a significant biological COD removal (= 80%). Pretreatment employing the O3/H2O2 process not only decreased the ultimate biodegradability of PPG effluent but also increased its inhibitory effects on activated sludge treatment speculatively due to the formation of less biodegradable oxidation by-products.

Effects of N‐heterocyclic polyaromatic hydrocarbons on survival, reproduction, and biochemical parameters in Daphnia magna

N-heterocyclic polycyclic aromatic hydrocarbons (N-PAHs) belong among newly identified classes of environmental pollutants with relatively high toxic potential. N-PAHs have been detected in air, soil, marine environments, and freshwater sediments. The N-PAHs are present at lower concentrations than their nonsubstituted analogues but their greater solubility would lead to greater bioavailibity and potential for toxic effects. Here we present results of acute and chronic toxicity in traditional aquatic invertebrate ecotoxicological model (Daphnia magna) along with assessment of biochemical responses. Studied biomarkers in D. magna exposed to N-heterocyclic derivatives included glutathione levels and activities of detoxication and antioxidative enzymes glutathione S-transferase and glutathione peroxidase. Phenanthrene and 1,10-phenathroline were the most toxic of all tested compounds (EC50 < 6 microM after 48 h exposure) and all tested N-PAHs suppressed reproduction of Daphnia magna. The data suggest that N-PAHs can induce oxidative stress in D. magna. The significant decline of glutathione content was found in animals treated with acridine, 1,10-phenanthroline, benzo(h)quinoline, phenantridine, and phenazine. Significant decrease of GPx activities relative to controls was found for all tested compounds except of phenanthrene and phenazine. Activities of GST increased after exposure to phenanthridine, phenazine, and benzo(h)quinoline, and declined in D. magna treated with phenanthrene (significant at one concentration) or anthracene (not significant). Our results confirmed significant acute as well as chronic toxicities of N-PAHs as well as potential of biochemical parameters to be used as early warning signals of toxicity in Daphnia magna.

Sequential anaerobic/aerobic biological treatment of olive mill wastewater and municipal wastewater

AbstractThis work investigated the efficiency of the combined anaerobic/aerobic biological co‐treatment of olive mill wastewater and primary municipal wastewater. A laboratory‐scale (6.5 L) upflow anaerobic sludge bed reactor received a mixture of olive mill wastewater and primary municipal wastewater at a loading rate ranging between 3 and 7 kg chemical oxygen demand (COD) m−3 day−1. The input COD concentration ranged between 1800 and 4400 mg L−1. The anaerobic reactor was operated at mesophilic conditions (35 °C). The effluent organic load was between 400 and 600 mg COD L−1, while the suspended solids removal efficiency varied between 75 and 95%. Average biogas production ranged between 3 and 4 L g−1 COD removed. The anaerobic reactor effluent was further treated in a laboratory‐scale activated sludge treatment plant. Aerobic treatment reduced the organic load even further to 85–175 mg COD L−1. However, the final effluent still retained a significant level of colour. Removal of colour was possible by ozonation or coagulation. Finally, the treated effluent was non‐ecotoxic, as indicated by the Daphnia magna toxicity test. This treatment method showed that it is feasible to treat olive mill wastewater in a municipal wastewater treatment plant by means of a high‐rate anaerobic reactor located between the primary clarifier and the aeration tank. Copyright © 2006 Society of Chemical Industry

Effects of copper pre-exposure routes on the energy reserves and subsequent copper toxicity inDaphnia magna

The hypothesis was tested that copper uptake routes affect the tolerance of Daphnia magna to copper and influence the energy reserves. These were determined in D. magna juveniles that had been exposed for 4 days to water borne and/or dietary copper (algae Pseudokichneriella subcapitata loaded with copper) at nominal concentrations of 0, 10, and 100 nM. Tolerance increased with dietary copper pre-exposure reflected in 24 and 48 h LC50 values of 466 and 398 nM at 100 nM pre-exposure versus 301 and 254 nM in controls, respectively. Control animals (no copper added to their exposure medium and diet) had the lowest lipid content and consequently the lowest energy content. The current study stresses the importance of addressing dietary exposure routes in metal toxicity assessments.