Ex situ remediation of contaminated sediments using mineral additives: assessment of pollutant bioavailability with the Microtox solid phase test

The aim of this work is to assess the potential ecotoxicological effects of contaminated sediments treated with mineral additives. The Microtox solid phase test was used to evaluate the effect of mineral additives on the toxicity of sediment suspensions. Four Mediterranean port sediments were studied after dredging and bioremediation: Sample A from navy harbor, sample B from commercial port and samples C and D from pleasure ports. Sediment samples were stabilized with three mineral additives: hematite, zero-valent iron and zeolite. Results show that all studied mineral additives can act as stabilizer agent in highly contaminated sediments (A and C) by decreasing dissolved metal concentrations and sediment toxicity level. On the contrary, for the less contaminated samples (B and D) hematite and zeolite can provoke toxic effect towards Vibrio fischeri since additive particles can favor bacteria retention and decrease bioluminescence emission.     

Arsenic in marine sediments from French Mediterranean ports: Geochemical partitioning,bioavailability and ecotoxicology

This work investigates arsenic mobility, bioavailability and toxicity in marine port sediments using chemical sequential extraction and laboratory toxicity tests. Sediment samples were collected from two different Mediterranean ports, one highly polluted with arsenic and other inorganic and organic pollutants (Estaque port (EST)), and the other one, less polluted, with a low arsenic content (Saint Mandrier port (SM)). Arsenic distribution in the solid phase was studied using a sequential extraction procedure specifically developed for appraising arsenic mobility in sediments. Toxicity assessment was performed on sediment elutriates, solid phases and aqueous arsenic species as single substance using the embryo-toxicity test on oyster larvae (Crassostrea gigas) and the Microtox test with Vibrio fischeri. Toxicity results showed that all sediment samples presented acute and sub-chronic toxic effects on oyster larvae and bacteria, respectively. The Microtox solid phase test allow to discriminate As-contaminated samples from the less contaminated ones, suggesting that toxicity of whole sediment samples is related to arsenic content. Toxicity of dissolved arsenic species as single substance showed that Vibrio fischeri and oyster larvae are most sensitive to As(V) than As(III). The distribution coefficient (Kd) of arsenic in sediment samples was estimated using results obtained in chemical sequential extractions. The Kd value is greater in SM (450 L kg^?1) than in EST (55 L kg^?1), indicating that arsenic availability is higher for the most toxic sediment sample (Estaque port). This study demonstrates that arsenic speciation play an important role on arsenic mobility and its bioavailability in marine port sediments.     

Evaluation of time-to-effects as a basis for quantifying the toxicity of contaminated sediments

Due to uncertainties as to appropriate procedures and dilution materials, most sediment tests are conducted only with undiluted, whole samples. Hence, it is not possible to use conventional concentration-response approaches to quantify toxicity of samples that elicit a 100% effect (e.g., mortality) at a preset test interval (typically 10 d). An alternative approach to quantifying the relative toxicity of test sediments is to determine time-to-effects. The objective of this study was to assess the utility of a time-to-effects approach for quantifying toxicity of freshwater sediments to the invertebrates Hyalella azteca and Chironomus tentans. Survival of both species and growth of C. tentans was determined using five sediments (four test samples and a control sediment) by destructively sampling replicate test chambers over the course of a "standard" 10-d assay. Studies with the control sediment and a non-toxic test sample indicated excellent recovery of test animals, even early in the test (e.g., <24 h) when individuals of both species are relatively small. Reasonable, typically monotonic, time-to-death relationships were observed for both H. azteca and C. tentans exposed to three comparatively toxic test sediments, all of which caused significant mortality by 10 d. Use of the time-to-effects approach allowed expression of toxicity of the three samples relative to one another, as well as documentation of decreases in toxicity of one of the sediments with increased storage time. These studies demonstrate the feasibility of use of time-to-effects as a basis for quantifying the relative toxicity of contaminated sediments.     

Direct comparison of amphipod sensitivities to dredged sediments from Spanish ports

The sensitivity of the benthic amphipod species Ampelisca brevicornis and Corophium volutator to dredged sediments was compared through simultaneous testing on the standard 10 days sediment toxicity test. The results of mortality obtained for 22 harbor sediments sampled at several Spanish ports were studied together with the physico-chemical characteristics of the samples to obtain the incidence of toxicity in terms of dredged material categories and to identify possible differences in the amphipod mortality results when using one or another test species. The results showed a similar incidence of toxicity for medium-high and highly contaminated sediments for both amphipod species, similar to that obtained through the comparison of the chemical concentrations measured in sediments with the single limit values used in Spain for dredged material characterization and management. On the contrary, C. volutator presented a higher mortality and a higher incidence of toxicity when exposed to low and medium-low contaminated sediments, which may have been caused by the lower sensitivity of A. brevicornis when exposed to sediments from its natural environment. When compared to other amphipod species used for whole sediment toxicity assessment, both amphipod species used in this study reported slightly higher sensitivities although these differences could have been associated to the different set of chemical compounds considered when characterizing the sediment samples. In this sense, the amphipod mortality results were better predicted through the use of mean quotients than just by comparing the measured chemical concentrations with the single limit values used in Spain, which indicates that the toxic response of both species was caused by the cocktail of contaminants present in the sediments. Finally, the correlation analysis identified a higher association between A. brevicornis mortality and the metallic contaminants while C. volutator was more correlated with the organic micro-pollutants. Despite these differences, the results indicate that Ampelisca brevicornis can be used as test organism for dredged material characterization when enough individuals of other recommended species such as Corophium volutator are not available.     

Heavy metal contamination from mining sites in South Morocco: 1. Use of a biotest to assess metal toxicity of tailings and soils

Our work was conducted to investigate the heavy metal toxicity of tailings and soils collected from five metal mines located in the south of Morocco. We used the MetPAD biotest Kit which detects the toxicity specifically due to the heavy metals in environmental samples. This biotest initially developed to assess the toxicity of aquatic samples was adapted to the heterogeneous physico-chemical conditions of anthropogenic soils. Contrasted industrial soils were collected from four abandoned mines (A, B, C and E) and one mine (D) still active. The toxicity test was run concurrently with chemical analyses on the aqueous extracts of tailings materials and soils in order to assess the potential availability of heavy metals. Soil pH was variable, ranging from very acidic (pH 2.6) to alkaline values (pH 8.0-8.8). The tailings from polymetallic mines (B and D) contained very high concentrations of Zn (38,000-108,000 mg kg(-1)), Pb (20,412-30,100 mg kg(-1)), Cu (2,019-8,635 mg kg(-1)) and Cd (148-228 mg kg(-1)). Water-extractable metal concentrations (i.e., soil extracts) were much lower but were highly toxic as shown by the MetPAD test, except for soils from mines A, E and site C3 from mine C. The soil extracts from mine D were the most toxic amongst all the soils tested. On this site, the toxicity of soil water extracts was mainly due to high concentrations of Zn (785-1,753 mg l(-1)), Cu (1.8-82 mg l(-1)) and Cd (2.0-2.7 mg l(-1)). The general trend observed was an increase in metal toxicity measured by the biotest with increasing available metal contents in tailings materials and soils. Therefore, the MetPAD test can be used as a rapid and sensitive predictive tool to assess the heavy metal availability in soils highly contaminated by mining activities.     

Effects of a chelating resin on metal bioavailability and toxicity to estuarine invertebrates: Divergent results of field and laboratory tests

Benthic invertebrates can uptake metals through diffusion of free ion solutes, or ingestion of sediment-bound forms. This study investigated the efficacy of the metal chelating resin SIR 300™ in adsorbing porewater metals and isolating pathways of metal exposure. A field experiment (Botany Bay, Sydney, Australia) and a laboratory toxicity test each manipulated the availability of porewater metals within contaminated and uncontaminated sediments. It was predicted that within contaminated sediments, the resin would adsorb porewater metals and reduce toxicity to invertebrates, but in uncontaminated sediments, the resin would not significantly affect these variables. Whereas in the laboratory, the resin produced the predicted results, in the field the resin increased porewater metal concentrations of contaminated sediments for at least 34 days and decreased abundances of four macroinvertebrate groups, and richness in all sediments. These contrasting findings highlight the limits of extrapolating the results of laboratory experiments to the field environment.     

Field validation of a battery of biomarkers to assess sediment quality in Spanish ports

Two marine invertebrates, the crab Carcinus maenas and the clam Ruditapes philippinarum, were used as bioindicator species to assess contamination when exposed in situ to sediment from different sites from four Spanish ports Cadiz (SW Spain), Huelva (SW Spain), Bilbao (NE Spain) and Pasajes (NE Spain). In an attempt to determine sediments toxicity, a combination of exposure biomarkers was analyzed in both species: metallothionein-like-proteins (MTLPs), ethoxyresorufin O-deethylase (EROD), glutathione S-transferase activity (GST), glutathione peroxidase (GPX) and glutathione reductase (GR). In parallel, physical and chemical characterization of the different sediments was performed and biological responses related to the contaminants. Significant induction of MTLPs was observed when organisms were exposed to metal contaminated sediments (port of Huelva), and EROD and GPX activities after exposure to sediments containing organic compounds (port of Bilbao and Pasajes). No significant interspecies differences were observed in biomarker responses except for the GST and GR.     

Evaluation of the phytotoxicity of contaminated sediments deposited "on soil": II. Impact of water draining from deposits on the development and physiological status of neighbouring plants at growth stage

As part of a study of the phytotoxic risk of spreading contaminated sediments "on soil", a laboratory experiment was carried out to assess the impact of water draining from sediments on peripheral vegetation. Drainage water was obtained in the laboratory by settling three sediments with different pollutants levels, and the supernatant solutions (respectively A1, B1, C1 drainage waters) were used as soaking water for maize (Zea ma?s L.) and ryegrass (Lolium perenne L.). The physicochemical characteristics of the supernatant water, particularly metal contents, showed a pattern of contamination, with C1>A1>B1. The plants tested were grown on soil for 21 days, before being soaked for another 21-day period with drainage water (treatments) and distilled water (control). Biomass parameters (fresh weight, length, etc.), enzymatic activity [glutamine synthetase (GS), phosphoenolpyruvate carboxylase (PEPc)] and Zn, Cu, Cd and Cr contents were measured on both the shoots and roots of each plant. Biomass parameters were stimulated by C1, not affected by A1 and decreased with B1 for maize, whereas they increased for ryegrass in all the treatments. Compared to the control, GS activity was stimulated by C1 in the shoots of both plants and inhibited by treatments B1 and C1 in maize roots. PEPc activity in ryegrass was 1.5-5 times higher with contaminated water treatment, while contrasting effects were observed in maize plants. Both plants showed greater accumulation of chromium and zinc than cadmium and copper. Treatment A1 was found to be less active on plant growth and have a lower impact on the physiological status (enzymatic activities) of both plants. Treatment C1 stimulated the growth and physiological status of the plants, especially in shoots, with higher metal accumulation values in both plants. Treatment B1 was found to show more variable effects on growth indices, enzymatic activity and metal accumulation according to plant species.     

Development and application of a sediment toxicity test using the benthic cladoceran Chydorus sphaericus

This study reports on the development and application of a whole sediment toxicity test using a benthic cladoceran Chydorus sphaericus, as an alternative for the use of pelagic daphnids. A C. sphaericus laboratory culture was started and its performance under control conditions was optimised. The test was firstly validated by determining dose-response relationships for aqueous cadmium and copper and ammonia, showing a sensitivity of C. sphaericus (96 h LC(50) values of 594 microg Cd/L, 191 microg Cu/L and 46 mg ammonia/L at pH 8) similar to that of daphnids. Next, sediment was introduced into the test system and a series of contaminated sediments from polluted locations were tested. A significant negative correlation between survival and toxicant concentrations was observed. It is concluded that the test developed in the present study using the benthic cladoceran C. sphaericus is suitable for routine laboratory sediment toxicity testing.     

Evaluation of the phytotoxicity of contaminated sediments deposited "on soil". I. Impact of water draining from the deposit on the germination of neighbouring plants

As part of a study of the phytotoxic risk of spreading of contaminated sediments "on soil", we carried out a laboratory experiment assessing the impact of water draining from sediments in a deposit scenario on the peripheral vegetation. The plant tested were the Chinese cabbage (Brassica campestris L. var. chinensis), maize (Zea mays L.) and ryegrass (Lolium perenne L.). The draining water samples (A1, B1 and C1) were obtained after decantation in laboratory of the sediments. The classification of the water sampled in decreasing order of cumulative contamination was C1 > A1 > B1. The B1 and C1 water samples inhibited the germination of seeds tested to various extents. The percentage of seeds that did not germinate was 1.3-fold times higher for Chinese cabbage with B1 than for the control and 2.3-fold times higher for ryegrass with C1 than for the control. Seeds watered with B1 had lower moisture contents than the control: 10% lower for ryegrass and maize and 50% lower for Chinese cabbage. An increase (about 1.5 microg/ml) in total soluble protein (versus the control) was observed for all three plant species tested in the presence of C1. Glutamine synthetase activity was significantly (1.35 times) higher in ryegrass seeds in the presence of C1 than in the control. We also observed changes in the specific activity of phosphoenolpyruvate carboxylase, which increased for ryegrass and decreased for maize as the concentration of contaminants in the water increased. The results show (i) the necessity to use different plant species to evaluate the toxic effect of sediment deposited on soil on the vegetation, and (ii) that soon as on germination an evaluation of an impact is possible.     

Can behavioural responses of Lumbriculus variegatus (Oligochaeta) assess sediment toxicity? A case study with sediments exposed to acid mine drainage

The São Domingos mine (Portugal) is, potentially, a good site for ecotoxicological studies, due to a pH and metal gradient of acid mine drainage. In this study, the toxicity of several mine sediments was evaluated using the aquatic oligochaete Lumbriculus variegatus as a test organism. Our hypothesis was that exposure to contaminated sediments would cause behavioural early warning responses in L. variegatus. Five sites, with pH ranging from 2.5 to 6.5, and with associated metals, were investigated. The results showed poor sediment quality in most of the collected sediments and Fe, S and As were the dominant elements in the samples. High mortalities were observed, ranging from 32.6 to 100%, indicating severe contamination. The collected sediments did not support good L. variegatus growth and significantly changed its behaviour. Early warning responses consisted of decreased locomotion and decreased peristaltic movements. A behaviour inhibition will affect the ecosystem balance by limiting the organisms' ability to avoid capture, which leads to a higher risk of predation.     

Influence of storage time on toxicity of freshwater sediments to benthic macroinvertebrates

Guidance concerning recommended storage times for sediments to be used in toxicity tests generally has not been based upon systematically collected experimental data. The objective of this study was to better define the effects of storage time on toxicity of a series of freshwater sediments. Sixteen sediments with varying types of contaminants were collected, homogenized and stored at 4 degrees C in 1 liter aliquots, which were periodically tested for toxicity to the amphipod Hyalella azteca and the midge Chironomus tentans after storage times of up to 101 weeks. The sediments ranged from non-toxic to extremely toxic (100% mortality) in 10-day assays, with several of the samples displaying an intermediate degree of toxicity (e.g. partial mortality, reduced growth). Biological responses in most of the samples did not vary with time relative to their statistical relationship to control values; samples identified initially as toxic (or non-toxic) tended to remain toxic (or non-toxic) regardless of when they were tested. The variations that were observed in biological responses over time generally were not systematic; that is, there were no apparent trends in samples becoming more (or less) toxic in the 10-day assays. This suggests that the source of at least some of the temporal changes in toxicity were due to inherent biological variability of the assays used to assess the sediments, rather than the effects of storage. In C. tentans tests with the least toxic sediments, among-replicate variability tended to be greater in initial assays than in tests with samples that had been stored for some period of time. This may have been due to the presence of indigenous competitive or predatory organisms that did not survive during prolonged storage.     

Passive samplers versus surfactant extraction for the evaluation of PAH availability in sediments with variable levels of contamination

The purpose of this study was to test the efficiency of passive solid samplers, polyoxymethylene (POM) strips and polydimethylsiloxane (PDMS) silicon tubing, to predict the bioavailability of native PAHs in contaminated sediments. Results were compared with worm bioaccumulation data and solid/liquid extraction using the surfactant Brij((R)) 700 (B700). The two passive samplers were found to act differently. The PDMS sampler overestimated the availability of PAHs in all studied sediments. The POM method provided results in accordance with those obtained with the B700 extraction. However, POM and B700 methods underestimated PAH availability in low contaminated sediments where biological factors (digestible organic matter) become important. Bioavailability of total PAHs was correctly predicted by POM and B700 in highly contaminated aluminum smelter sediments. A closer examination of individual PAH results indicated that both techniques overestimated the availability of large molecules with logK(ow)>6 suggesting a biological mechanism limiting uptake of larger PAHs which seems to be related to the molecular size of compounds.     

化学物质对发光菌的联合毒性评价方法

毒性单位法(TU)的理论基础来源于剂量加和模型(DA),目前仅在二元联合毒性评价中广泛应用。为了确定TU模型适合评价的混合物类型,实验选取5种剂量效应曲线类型不同的物质,采用微板光度计测试了一元、二元混合物对发光菌青海弧菌-Q67(Vibrio-qinghaiensis sp.-Q67)的急性毒性。根据物质的剂量效应曲线形状将物质分为A、B、C 3类,利用毒性单位法(TU)和联合作用定义法分别对AA类、AB类、AC类、BC类混合物进行分析。结果表明,TU法仅适合于由剂量效应曲线接近直线的物质组成的混合物进行联合毒性的评价。以效应为基准、TU模型为框架建立了TU’模型,该模型可以满足对任何类型已知成分的混合物或者未知成分的实际水样之间的多元联合作用的评价。     

Comparison of in-house-developed ELISA with HPLC techniques for the analysis of atrazine residues

In-house developed ELISA was standardized to monitor atrazine residues in different environmental samples. The standard curve was linear, indicating an increase in log concentration with decrease in absorbance (%B/B(0)=1.075-0.042 Log C; r= -0.966). The middle of the test was at 75 ng/L and the lowest detection limit at 4 ng/L. ELISA significantly correlated with the high performance liquid chromatography (HPLC) (r=0.990). Internal validation showed good accuracy and precision. Maximum atrazine residues were present in Jehlum River water/sediments and maize/sugarcane plant roots. Most of the food samples were found to be contaminated. ELISA required less clean-up steps than HPLC, but showed matrix effect in soil/colored extracts.     

Potential impacts of water injection dredging on water quality and ecotoxicity in Limehouse Basin, River Thames, SE England, UK

The use of water injection dredging (WID) is increasing in the UK's inland waterways and marinas. Jets of water are injected under low pressure directly into bottom sediment creating a turbulent water-sediment mixture that flows under the influence of gravity. Many of these sediments are highly contaminated and little is known of the effects of contaminant release on water quality or the risk to biota living in both the sediment and the water column. Sediment cores were collected from Limehouse Basin, a proposed WID site in SE England and current sediment toxicity was assessed using a number of techniques. Comparison of metal data to US sediment quality guidelines indicated intermediate levels of toxicity while, calculation of acid volatile sulphide to simultaneously extracted metal ratios underestimated the potential toxicity to sediment dwelling organisms. In contrast, porewater ammonia concentrations were in excess of all published ecotoxicological guidelines and indicate serious risk to biota. Re-suspension experiments were used to mimic the effects of WID on overlying water quality and ecotoxicity tests were carried out on elutriates using Daphnia magna to examine the impacts on biota. Concentrations of a range of metals in the elutriates predict that adverse biological effects would be observed during WID, however only 10% of the elutriate samples caused an adverse effect on Daphnia. Limehouse Basin is a complex aquatic environment receiving predominantly fresh waters while the sediments have high porewater chloride concentrations reminiscent of previous tidal inputs to the basin, making the choice of test organism problematic.     

Effects of storage method and duration on the toxicity of marine sediments to embryos of Crassostrea gigas oysters

The objective of laboratory sediment bioassays is to estimate in situ toxicity. This goal is difficult to achieve, as one of the main limitations of sediment toxicity tests is disruption of sediment geochemistry during sampling, handling and preservation. The effects of storage on the estimation of marine sediment toxicity to Crassostrea gigas embryos and larvae were investigated. Three storage methods and four storage periods were compared with three different sediment types contaminated by heavy metals, polycyclic aromatic hydrocarbons (PAHs) and both contaminants. Freezing and freeze-drying considerably increased the toxicity of decanted sediments and their elutriates as compared to the toxicity obtained with fresh sediments. Concerning the elutriates, the toxicity found with frozen and freeze-dried sediments was correlated with DOC, ammonia and PAH contents. However, the toxicity of fresh sediments kept at 4 degrees C increased with increasing duration of storage and was also correlated with the amount of ammonia in the elutriates.     

Seasonal variation of sediment toxicity in the Rivers Dommel and Elbe

Contaminated sediment in the river basin has become a source of pollution with increasing importance to the aquatic ecosystem downstream. To monitor the temporal changes of the sediment bound contaminants in the River Elbe and the River Dommel monthly toxicity tests were applied to layered sediment and river water samples over the course of 10 months. There is an indication that contaminated sediments upstream adversely affected sediments downstream, but this process did not cause a continuous increase of sediment toxicity. A clear decrease of toxic effects in water and upper layer sediment was observed at the River Elbe station in spring related to high water discharge and algal blooms. The less obvious variation of sediment toxicity in the River Dommel could be explained by stable hydrological conditions. Future monitoring programmes should promote a more frequent and intensive sampling regime during these particular events for ecotoxicological evaluation.     

Accumulation and histopathological damage in the clam Ruditapes philippinarum and the crab Carcinus maenas to assess sediment toxicity in Spanish ports

The degree of contamination and toxicity in sediment from four Spanish ports (Cádiz, Huelva, Pasajes and Bilbao) was assessed in the present study. Two marine invertebrate species, the shore crab Carcinus maenas and the clam Ruditapes philippinarum, were exposed to the different sediments under laboratory conditions for 28 days. Relationships were developed among metal contamination (As, Cd, Cr, Cu, Fe, Hg, Mn, Ni, Pb and Zn) in sediments, metal accumulation in gill tissues and histopathological lesions in different biological tissues to assess sediment toxicity. A multivariate analysis approach was used to calculate sediment quality guidelines (SQGs) by linking metal concentration in sediments with histopathological lesions measured in the exposed organisms. The results showed significant contaminant accumulation and histopathological lesions due to As, Pb and Zn in sediments at the port of Huelva; As, Cr, Cu, Hg and Ni at the ports of Pasajes and Cádiz; and Cr and Hg at the port of Bilbao. The link between chemical concentration in sediments and histopathological lesions allowed the determination of SQGs for the chemicals As, Cd, Cu, Cr, Hg, Ni and Zn.     

Assessing sediment toxicity and arsenite concentration with bacterial and traditional methods

Three sediment samples LP (pool where logs are stored), LF (brook through landfill area), KN (Kaskesniemi) which is in Lake Pyh?selk? downstream from the mill, were taken from an old sawmill area and one from the unpolluted Lake H?yti?inen. The arsenite concentration was measured by GFAAS and two arsenite biosensing bacterial strains Pseudomonas fluorescens OS8 (pTPT31) and Escherichia coli MC1061 (pTOO31). The toxicity of sediment and pore water samples was determined by using luminescent bacteria (Flash test) and, further, whole sediment toxicity was measured using 10 days growth test and 50 days emergency test with midges (Chironomus riparius). With the flash test a lowered EC50 value was found only in sediment LF (EC50=0.17 v/v%). The Flash test indicated that all sediment samples taken from the sawmill area were highly toxic to bacteria, whereas growth and the emergence of chironomids showed no effects in other samples than LF. The midges tolerate well the contaminated environment. In contrast, bioavailability of arsenite of sediment samples KN and LF was quite high determined using the biosensor-strains in a direct contact assay. The bioavailable fraction of sediment LP was 6-10% out of the total arsenite concentration obtained with GFAAS (0.46-0.77 microg g-1 dw). The results show that the choice of analysis method grossly affects the outcome without any of the method giving an incorrect result. Different methods measure different parameters of a toxic sample and can thus be used to complement each other.