Toxicity identification in metal plating effluent: Implications in establishing effluent discharge limits using bioassays in Korea

Because of complexity and diversity of toxicants in effluent, chemical analysis alone gives very limited information on identifying toxic chemicals to test organisms. Toxicity identification evaluation (TIE) techniques have been widely used to identify toxicants in various samples including industrial wastewater as well as natural waters. In response to new regulation for effluent discharge in Korea, which will be effective from 2011, a necessity of studies emerges that investigates toxicity levels in industrial effluents. This work was a preliminary study examining toxicity levels in effluent from one metal plating factory using Daphnia magna (48 h immobility) and identifying toxicity-causing substances. Toxicity tests showed variability on different sampling occasions and the results of TIE methods indicated that both organic compounds and metals contributed to the observed toxicity in metal plating effluent. Further studies are necessary to help reduce effluent toxicity especially from direct dischargers, who will have to comply with the new regulation.     

Burrowing and avoidance behaviour in marine organisms exposed to pesticide-contaminated sediment

Behavioural effects of marine sediment contaminated with pesticides (6000 ppm parathion, 200 ppm methyl parathion, 200 ppm malathion) were studied in a number of marine organisms in laboratory tests and in situ. The burrowing behaviour in Macoma baltica, Cerastoderma edule, Abra alba, Nereis diversicolor and Scoloplos armiger was impaired in the contaminated sediment compared to control. The impairment was most pronounced in the laboratory tests, where almost no burrowing occured. In a very simple laboratory set-up, highly significant avoidance of the contaminated sediment was demonstrated for Crangon crangon and Solea solea, but not for Carcinus meanas and Pomatoschistus minutus. The validity of both behavioural tests was supported by in situ observations and investigations on the distribution of the species. It is concluded that both tests are useful tools in the assessment of the impact of contaminated sediments.     

Toxicity studies of elemental sulfur in marine sediments

Elemental sulfur(ES) is a component essential for proper development of animals, but it can be toxic for aquatic organisms. The objectives of the study reported here included determination of ES concentrations in sediment collected in the area of the Gulf of Gdansk(Baltic Sea) and search for the possible correlations with the ecotoxicity results. Sediment samples were collected from four locations: the Vistula River mouth, in the Port Basin, in the area of discharge of the treated wastewater from the Wastewater Treatment Plant(WWTP) and in the area of the sunken World War Ⅱ s/s 'Stuttgart' shipwreck. The levels of elemental sulfur were determined using a gas chromatograph coupled with a mass spectrometer(GC-MS). Toxicity of marine sediment samples was estimated for three biotest organisms-bioluminescent bacteria Vibrio fischeri, crustacean Heterocypris incongruens, and Synapis alba plant. The highest toxicity of sediments for all indicator organisms was observed in samples taken at the place of the shipwreck(2-100%). The same samples had a high sulfur content(16.7-143.2 μg/g dry weight(d.w.))therefore, it was decided to investigate whether the presence of ES in the sediment in the studied area can have an impact on the results of the ecotoxicity determination in real samples. However, the removal of sulfur from the samples resulted in no significant changes in the level of toxicity of the samples. In this research it could not be confirmed that the presence of elemental sulfur is the only factor responsible for the observed sediment toxicity. It seems that other compounds or their mixtures present in the sediment may have a significant influence on the results obtained.     

The acute toxicity of motor fuels to brackish water organisms

The addition of synthetic additives, such as methanol and methyl tertiary butyl ether (MTBE) to the base fuel, with the aim of raising the octane content of motor fuel, is currently of special interest in Sweden. A question, which has to be solved, is whether it is preferable to carry the additives by ship in separate tanks, because of the probable enhanced toxicity of the motor fuels by the addition of solvents, or not. The acute toxicity of these blend fuels has been compared with the acute toxicity of base fuel, gasoline and low leaded gasoline. As test organisms, two representative species of the Baltic Sea have been used, the harpacticoid Nitocra spinipes and the bleak, Alburnus alburnus. The results of the tests show that the acute toxicity of base fuel to aquatic organisms is not increased by the addition of 15% methanol or 5% MTBE.     

Evaluation of induction of metallothionein-like proteins (MTLPs) in the polychaetes for biomonitoring of heavy metal pollution in marine sediments

Polychaetes are suitable organisms for evaluation of impact of sediment pollution. We evaluated toxicity of cadmium and copper and measured metallothionein-like proteins (MTLPs) in the polychaete Perinereis nuntia. At the same concentration ranges copper was unexpectedly more toxic than cadmium. Copper also caused no significant increase in MTLPs in the polychaetes. When P. nuntia and another polychaete species, Cirratulus cirratus were cultured in the contaminated sediments collected from Lake Sihwa (Korea), a high mortality of 80% was observed on day 6 in P. nuntia in the sediment with the highest metal concentration. However, no mortality was observed up to 35 days in C. cirratus in any sediment. MTLP contents between two species also varied. These findings suggest that MTLP induction response in the polychaetes varies with the metal type and species and it may be used as a biomarker of sediment pollution in the polychaetes after further validation and field trials.     

Validation of Arenicola marina in field toxicity bioassays using benthic cages: biomarkers as tools for assessing sediment quality

Sediment toxicity assessments using caged organisms present advantages over using laboratory and native community studies. The use of caged Arenicola marina in sediment toxicity assessments was evaluated. Lugworms were exposed in situ to sediments from coastal and port areas in Spain for seven days, and the activities of the biotransformation enzymes ethoxyresorufin O-deethylase, dibenzylfluorescein dealkylase and glutathione S-transferase, the activities of the antioxidant enzymes glutathione reductase and glutathione peroxidase and lipid peroxidation were then analyzed as biomarkers. Biomarker results and sediment physicochemical data were integrated. Cádiz Bay (SW Spain) sediments presented metal contamination that was not linked to a biochemical response. In LPGC Port (SW Spain), Pb contamination exhibited a moderate toxic potential, while PAHs, and presumably pharmaceuticals, provoked biochemical responses that efficiently prevented lipid peroxidation. In Santander Bay (N Spain), exposure to PAHs and, presumably, pharmaceuticals induced biomarker responses, but lipid peroxidation occurred nevertheless. These results indicated that caged A. marina were effective for the assessment of sediment quality and that the selected biomarkers were sufficiently sensitive to identify chemical exposure and toxicity.     

Comparative performances of eggs and embryos of sea urchin (Paracentrotus lividus) in toxicity bioassays used for assessment of marine sediment quality

The potential toxicity of sediments from various ports was assessed by means of two different liquid-phase toxicity bioassays (acute and chronic) with embryos and eggs of sea urchin Paracentrotus lividus. Performances of embryos and eggs of P. lividus in these bioassays were compared for their interchangeable applicability in integrated sediment quality assessment. The obtained endpoints (percentages of normally developed plutei and fertilized eggs) were linked to physical and chemical properties of sediments and demonstrated dependence on sediment contamination. The endpoints in the two bioassays were strongly correlated and generally exhibited similar tendency throughout the samples. Therein, embryos demonstrated higher sensitivity to elutriate exposure, compared to eggs. It was concluded that these tests could be used interchangeably for testing toxicity of marine sediments. Preferential use of any of the bioassays can be determined by the discriminatory capacity of the test or vulnerability consideration of the test subject to the surrounding conditions.     

Effects of new antifouling compounds on the development of sea urchin

Tributyltin oxide (TBTO) has been used worldwide in marine antifouling paints as a biocide for some time. However, it produced toxic effects, especially in marine water/sediment ecosystems. Consequently, its use in antifouling paints has been prohibited in many countries. In this study, the toxicity of alternative and/or new antifouling biocides compared with TBTO is assessed by a biological method. The effects of these chemicals on marine species have not been well studied. This paper assesses, comparatively, the effects of eight biocides on sea urchin eggs and embryos. The chemicals assessed were TBTO, Irgarol 1051, M1 (the persistent degradation product of Irgarol), Diuron, zinc pyrithione, 'KH101', 'Sea-Nine 211', and copper pyrithione. For these chemicals, toxicity appears to be in the order zinc pyrithione > Sea-Nine 211 > KH101 > copper pyrithione > TBTO > Diuron approximately = Irgarol 1051 > M1. Here, we show that zinc pyrithione, Sea-Nine 211, KH101, and copper pyrithione are much more toxic to sea urchins than TBTO or the other chemicals.     

Distribution, magnitude and characterization of the toxicity of Ukrainian estuarine sediments

During the Soviet era, Ukraine was an important industrial and agricultural region of the Soviet Union. This industrial and agricultural activity resulted in contamination of Ukraine’s estuaries with legacy anthropogenic pollutants. Investigations on the toxicological effects of this estuarine contamination have been limited. For this research, we measured the toxicity of contaminated sediments from four Ukrainian estuaries to several aquatic organisms over 3 years. Sediment chemical analyses and whole sediment toxicity identification evaluations (TIEs) were also performed to determine the classes of contaminants contributing to toxicity. Toxic sediments were observed in several of the Ukrainian estuaries and chemical analyses of the sediments demonstrated anthropogenic contaminants were widely distributed. Contaminants were also detected in macrobenthic organisms collected from the sediments. Several lines of evidence, including TIEs, indicated hydrophobic organic chemicals (HOCs) were contributing substantially to observed toxicity. This information can guide environmental managers to prioritize portions of the estuaries requiring remediation.     

Axenic Cultures of Tetrahymena Pyriformis as Toxicological Tools

The method of determining toxicity in streams by the use of axenic cultures of the protozons species Tetrahymena pyriformis is described. Contrary to the similar tests which are used in medicine in defined but rather complicated culture media, for the toxicological experiments in surface water it is possible to use very simple media for the cultivation of test organisms. Duration of the whole direct test of toxicity is limited to 24 hours. The other advantages of using the test organism Tetrahymena pyriformis are mentioned as well. According to the agricultural and industrial pollution which often takes place in our surface waters five chemicals, most of them pesticides, were tested.     

Toxicity of sediment-incorporated drilling fluids

The 24, 96, or 168h LC₅₀s of four used drilling fluids or barite incorporated into sediment were determined in toxicity tests with lancelets (Branchiostoma caribaeum), a benthic chordate. Concurrent to LC₅₀ determinations, EC₅₀s were calculated from the number of lancelets that did not burrow into contaminated sediments. Observations of the burrowing behaviour allowed quantitation of effects after 24h exposures to each of the drilling fluids whereas lancelet mortality was sufficient to calculate 24h LC₅₀s for only one drilling fluid. Drilling fluids were less toxic to lancelets when incorporated into sediments than to mysids (Mysidopsis bahia) or benthic invertebrate communities in water-column exposures.     

Using growth measures in the freshwater shrimp Caridina nilotica as biomarkers of Roundup® pollution of South African freshwater systems

There has been global concern about the effect of toxic chemicals on aquatic biota due to the upsurge in contamination of aquatic ecosystems by these chemicals, which includes pesticides. Roundup® and other glyphosate-based herbicides are frequently used in the chemical control of weeds and invading alien plant species in South Africa. These bio-active chemicals ultimately get into water courses directly or indirectly through processes such as drifting, leaching, surface runoff and foliar spray of aquatic nuisance plants. However, there is no South African water quality guideline to protect indigenous freshwater non-target organisms from the toxic effects of glyphosate-based herbicides. This study evaluated the possible use of growth measures in Caridina nilotica as biomarkers of Roundup® pollution as part of developing glyphosate water quality guideline for the protection of aquatic life in South Africa. Using static-renewal methods in a 25-day growth toxicity test, 40 days post hatch shrimps were exposed to different sub-lethal Roundup® concentrations of 0.0 (control), 2.2, 2.8, 3.4, 4.3 and 5.4 mg/L. Shrimps were fed daily with TetraMin® flake food and test solutions changed every third day. Shrimp total lengths and wet weights were measured every fifth day. These data were used to determine the shrimp’s growth performance and feed utilization in terms of percent weight gain (PWG), percent length gain (PLG), specific growth rate (SGR), condition factor (CF), feed intake (FI), feed conversion ratio (FCR) and feed conversion efficiency (FCE). Moulting was observed for 14 days and the data used to determine the daily moult rate for each concentration. Results of growth performance and food utilization indices showed that growth was significantly impaired in all exposed groups compared to control (p < 0.05). Moulting frequency was also higher in all exposed groups than in control (p < 0.05). Although all the tested growth measures proved to be possible biomarkers of Roundup® pollution, moulting frequency gives a clearer indication of the sub-lethal effects of Roundup® toxicity.     

Application of bioassays in toxicological hazard, risk and impact assessments of dredged sediments

Given the potential environmental consequences of dumped dredged harbour sediments it is vital to establish the potential risks from exposure before disposal at sea. Currently, European legislation for disposal of contaminated sediments at sea is based on chemical analysis of a limited number of well-known contaminants for which maximum acceptable concentrations, action levels (ALs), have been set. The present paper addresses the issue of the applicability of in vitro and in vivo bioassays for hazard, risk and local impact assessment of dredged polluted sediments to be disposed of at sea. It discusses how and to what extent selected bioassays can fill in the gaps left open by chemical analysis and the way in which the bioassays may contribute to the present licensing system for disposal. Three different purposes for application were distinguished: the most basic application (A) is a rapid determination of the hazard (potential toxicity) of dredged sediments which is then compared to ALs in a licensing system. As with chemical analysis on whole sediment extracts, the bioavailability of the chemicals is not taken into account. As in vitro assays with sediment extracts are not sensitive to matrix effects, a selection of specific in vitro bioassays can be suitable fast and standardized additions for the licensing system. When the outcome of (A) does not convincingly demonstrate whether the sediment is clean enough or too polluted, further bioanalysis can help the decision making process (B). More aspects of the mostly unknown complex chemical mixtures are taken into account, including the bioavailability and chronic toxicity focusing on ecologically relevant endpoints. The ecotoxicological pressure imposed by the dredged sediments can be quantified as the potentially affected fraction (PAF) based on chemical or biological analysis of levels of contaminants in sediment or biota. To validate the predicted risk, the actual impact of dumped harbour sediments on local ecosystems (C) can be determined using a dedicated set of in vitro and in vivo bioassays as well as bio-indicators selected based on the information obtained from (A) and (B) and on the characteristics of the local ecosystem. Conversely, the local sediment impact assessment (C) can direct fine-tuning of the selection of chemical and bioassay analyses and for setting safe levels in the licensing system. It is concluded that in vitro and in vivo bioassays and biological indicators are useful tools in the process of hazard, ecotoxicological risk and impact assessment of dredged harbour sediments, provided they are consciously chosen and quality criteria for assay performance are defined.     

Pore water testing and analysis: the good,the bad,and the ugly

The increasingly common practice of collecting and assessing sediment pore water as a primary measure of sediment quality is reviewed. Good features of this practice include: pore water is a key exposure route for some organisms associated with sediments; pore water testing eliminates particle size effects; pore water analyses and tests can provide useful information regarding contamination and pollution. Bad features include: pore water is not the only exposure route; pore water tests lack chemical or biological realism: their "sensitivity" relative to other tests may be meaningless due to manipulation and laboratory artifacts; many sediment and surface dwelling organisms are not directly influenced by pore water. Bad features can become ugly if: other exposure pathways are not considered (for toxicity or bioaccumulation); manipulation techniques are not appropriate; pore water tests are inappropriately linked to population-level effects. Pore water testing and analyses can be effective tools provided their limitations are well understood by researchers and managers.     

Synergistic toxic effects of zinc pyrithione and copper to three marine species: Implications on setting appropriate water quality criteria

Zinc pyrithione (ZnPT) is widely applied in conjunction with copper (Cu) in antifouling paints as a substitute for tributyltin. The combined effects of ZnPT and Cu on marine organisms, however, have not been fully investigated. This study examined the toxicities of ZnPT alone and in combination with Cu to the diatom Thalassiosira pseudonana, polychaete larvae Hydroides elegans and amphipod Elasmopus rapax. Importantly, ZnPT and Cu resulted in a strong synergistic effect with isobologram interaction parameter λ > 1 for all test species. The combined toxicity of ZnPT and Cu was successfully modelled using the non-parametric response surface and its contour. Such synergistic effects may be partly due to the formation of copper pyrithione. It is, therefore, inadequate to assess the ecological risk of ZnPT to marine organisms solely based on the toxicity data generated from the biocide alone. To better protect precious marine resources, it is advocated to develop appropriate water quality criteria for ZnPT with the consideration of its compelling synergistic effects with Cu at environmentally realistic concentrations.     

Sediment PAH: Contrasting levels in the Caspian Sea and Anzali Wetland

A comparative study of 23 PAH congeners in sediment of the Caspian Sea coast and Anzali Wetland was conducted in 2010. Surface sediment was analyzed using chromatography and mass spectrometry. Total PAH concentrations ranged between 212 and 9009 ng g⁻¹ dw. Spatial distribution maps revealed that PAH levels were higher in the coastal areas of the Caspian Sea where oil related activities have been common since 1800’s. Diagnostic ratios analysis indicated that PAHs largely originated from petrogenic processes. PAH toxicity level was assessed using sediment quality guidelines and toxic equivalent concentrations to determine toxic effects on marine organism. Based on these investigations, in our study areas, the probability of toxicity for benthic organisms is “low to medium”. The toxic equivalent concentrations of carcinogenic PAHs varied between 11 and 231 ng TEQ/g; higher total toxic equivalent concentrations values were found in the coastal areas of the Caspian Sea.     

Toxicity identification in metal plating effluent: implications in establishing effluent discharge limits using bioassays in Korea

Because of complexity and diversity of toxicants in effluent, chemical analysis alone gives very limited information on identifying toxic chemicals to test organisms. Toxicity identification evaluation (TIE) techniques have been widely used to identify toxicants in various samples including industrial wastewater as well as natural waters. In response to new regulation for effluent discharge in Korea, which will be effective from 2011, a necessity of studies emerges that investigates toxicity levels in industrial effluents. This work was a preliminary study examining toxicity levels in effluent from one metal plating factory using Daphnia magna (48 h immobility) and identifying toxicity-causing substances. Toxicity tests showed variability on different sampling occasions and the results of TIE methods indicated that both organic compounds and metals contributed to the observed toxicity in metal plating effluent. Further studies are necessary to help reduce effluent toxicity especially from direct dischargers, who will have to comply with the new regulation.     

A comparative study of toxicity identification using Daphnia magna and Tigriopus japonicus: Implications of establishing effluent discharge limits in Korea

In Korea, the new permission criteria for industrial effluents based on Daphnia magna acute toxicity tests will be gradually implemented starting from 2011. Thus, in this study, toxicity assessment and identification using a marine species (Tigriopus japonicus) and the freshwater species (D. magna) was comparatively investigated. Effluent from an acid mine drainage treatment plant showed acute toxicity toward both organisms due to low pH, which was removed by neutralization of the effluent. Additionally, evaluation of the effluent of an electronics company revealed that Cu was attributable to the observed toxicity, and the effluent was more toxic toward T. japonicus than D. magna. Moreover, effluents from a metal plating factory were acutely toxic toward D. magna (6.50 TU), while they were not toxic against T. japonicus. Toxicity identification revealed that the high level of Cl⁻ (12,841 mg L⁻¹) was the cause of toxicity. Thus, the effluents had no effect on the marine species, T. japonicus. These findings suggest that a marine species rather than a freshwater species is more desirable for toxicity assessment of industrial effluent discharged into the saltwater, and thus should be considered in the legislation of toxicity-based discharge limits in Korea.     

Integrative assessment of coastal pollution: Development and evaluation of sediment quality criteria from chemical contamination and ecotoxicological data

Elutriate embryo-larval bioassays with sea-urchins (Paracentrotus lividus) were conducted concurrently with chemical analyses of sediments and biota as part of an integrative assessment of pollution in highly productive coastal regions. High metal contents and organic compounds in sediments and mussels were found in localised areas from the inner part of the estuaries indicating a clear anthropogenic influence. In particular, average maximum concentrations of 2803 mg Cu/kg dw, 776 mg Pb/kg dw, 2.5 mg Hg/kg dw and 5803 μg ∑₇PAHs/kg dw were measured in sediments from the most polluted sites. Significant correlations were observed between sediment chemistry and toxicity bioassays. Moreover, the Mantel test revealed a significant correlation (r_M=0.80; p<0.01) between sediment pollutant concentrations and toxicity data profiles. In addition, sediment quality criteria were used to help in the ecological interpretation of sediment chemistry data and to identify pollutants of concern. The toxicity bioassays identified polluted sites and quantified the level of toxicity, providing a cost-effective tool to complement the routine chemical monitoring currently conducted in European coastal waters with ecologically relevant information. This is in line with the recent European legislation that advocates the use of biological tools with the ultimate aim of protecting marine resources from anthropogenic substances that will affect their sensitive early life stages.     

Some effects of pollutants on the behaviour of the bivalve Tellina tenuis

Prediction of safe levels of effluent discharge have in the past been largely based on the results of short term acute toxicity tests (LC_{5 0}s). Such tests are usually carried out in unnatural experimental conditions, and in consequence the results are of only limited value. There is a need to enlarge the concept of the routine bioassay test to include quantitative measurement of the effects of pollutants on behaviour, physiology and metabolism. This paper describes a simple rapid bioassay test in which the effect of pollutants on the burrowing of the bivalve Tellina tenuis has been used as a quantitative measure of pollutant effect.