Simulating a heavy metal spill under estuarine conditions: Effects on the clam Scrobicularia plana

We describe the effect of heavy metals Zn, Cd, Pb and Cu on the induction of methallothioneins on the clam Scrobicularia plana along a salinity gradient simulated under laboratory conditions. The clams were exposed to constant heavy metal concentrations in a dynamic estuary simulator during a 15-day assay to investigate possible induction of metal-binding proteins in them. The concentration of heavy metals in water was analysed. Clams were analysed for methallothionein concentrations. The speciation of Zn, Cd, Pb and Cu along the salinity gradient was modelled. Zn showed the highest concentrations and its prevalent species was the free ion. Intersite differences have been observed in methallothionein concentration and related to the salinity gradient. It seems that synthesis of methallothioneins is the result of physiological forces acting in concert with the changes in the chemical speciation of metals, owing to the trace metals uptake is controlled by means of an interaction of physiology and physicochemistry.     

Potential toxicity of resuspended particulate matter and sediments: Environmental samples from the Bay of Banyuls-sur-Mer and Thermaikos Gulf

This contribution attempts to determine whether, and to what degree, environmental samples of resuspended particulate matter and sediments exert a toxicological impact. Further, an attempt is made also to screen the toxic level of potentially hazardous sites, based upon established sediment toxicity criteria. Therefore, a rapid, cost-effective and highly sensitive biotest (bioluminescence assay, based upon marine bacteria) has been applied on: biological fluid extracts; bottom sediments; and sediment trap samples. Samples were taken either from the Bay of Banyuls-sur-Mer (northwestern Mediterranean, France) or Thermaikos Gulf (northeastern Mediterranean, Greece). Biological fluid extracts and sediment trap samples corresponded to periods of resuspension events, or preceding and following such events.The results have revealed that the sampling strategy and biotest implemented in this study might be a useful tool for screening the toxicity of resuspended matter and sediments. Resuspension events appear to be able to exert an influence on the chemical forms of the micro-pollutants; thus on their bioavailability and toxicity. Nevertheless, based upon chemical analysis combined with the bioassay, the toxic level of the sediment samples could be determined, with the level at potentially hazardous sites being characterised.     

Application of the luciferase cell culture bioassay for the detection of refined petroleum products

A luciferase cell culture-based bioassay, developed to detect 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD)-like activity of halo-genated and polycyclic aromatic hydrocarbons, was optimized to detect refined petroleum products and to determine their relative inducing potency. Quality control standards from 32 refined products (gasolines and diesels, jet fuels, lubricating oils, fuel oils and weathered products) and three commercial products were evaluated. Induction equivalents (I-EQs) were determined by direct comparison of the EC50 and EC20 values (based on the median and 20% TCDD maximal response, respectively) from dose-response curves for each product to those obtained with TCDD. Most petroleum products were active in the luciferase bioassay, with those products composed of fractions produced later in the distillation process (i.e. fuel oils) inducing higher levels. Additionally, weathering of products reduced their induction potency. Based on the high I-EQ estimates of many products, biological effects associated with exposure may have been previously underestimated using other diagnostic methods.     

Expression of heat shock protein 70 in a permanent cell line (EPC) exposed to sediment extracts from the North Sea and the Baltic Sea

Heat shock proteins 70 (hsp70) are known to be induced by a great variety of chemical stressors. The effects of different environmental contaminants, which were identified in sediments of the North Sea and the Baltic Sea as main contaminants or as contaminants of special toxicological interest, on the extent of expression of hsp70 in a permanent cell line Epithelioma papulosum cyprini (EPC) were investigated. Concentrations of the single contaminants leading to elevated hsp70 levels were compared to the respective concentrations of these contaminants encountered in the sediment extracts thus allowing an assessment of their contribution to the toxicity of these sediments. Analysis of hsp70 was performed using western blotting techniques with subsequent comparative quantification by densitometry. Eleven of thirteen contaminants have shown a dose-dependent increase in hsp70 levels. An expression of hsp70 was observed in most cases between 0.1 and 10 microg/mL test solution. Some of the investigated stressors led to extremely high hsp70 contents even at low concentrations. However, in most cases contaminant concentrations in sediments were too low to cause an effect. For this reason, effects of the tested sediments are attributed to other contaminants or rather to mixtures of stressors.     

Some Advantages of the Duckweed Test to Assess the Toxicity of Environmental Samples

In Germany, wastewater legislation requires all municipal and industrial leachate to be subjected to toxicity tests. However, no phytotoxicity tests using higher plants are currently included among the standard tests. Freshwater microalgae have been used in most phytotoxicity tests and have often been considered as surrogates for higher plants. However, microalgae often do not show the same sensitivity as higher plants and have major disadvantages for the testing of unmodified environmental samples. In the following study, we evaluated the suitability of the giant duckweed Spirodela polyrhiza for assessing the toxicity of a municipal landfill leachate and two leachates of copper mining residue. Duckweed static toxicity tests were performed, and frond number was the endpoint used to calculate EC₅₀ values. Symptoms of stress (chlorosis, necrosis, root destruction, and colony breakup) were also recorded. The landfill leachate was toxic with EC_50;96h values ranging from 1.3 to 2.7% leachate (v/v). Toxicity of the copper slag leachate was largely determined by the elution method used. Leachate obtained using conventional German leaching methods (S4-eluate) was not toxic to duckweed, whereas EC₅₀ values for the pH_stat4-eluate ranged from 3.2 to 4.2% leachate (v/v). The results demonstrate the suitability of S. polyrhiza for the testing of unmodified wastewater samples and provide further evidence for the addition of a duckweed toxicity test to the standard tests conducted in Germany.     

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.     

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.     

Harmful algal bloom species and phosphate-processing effluent: field and laboratory studies

In 2002, the Florida Department of Environmental Protection began discharging phosphate-processing effluent into Bishop Harbor, an estuary within Tampa Bay. Because of concerns that the effluent would serve as a nutrient source for blooms of the toxic dinoflagellate Karenia brevis, a field monitoring program was established and laboratory bioassays were conducted. Several harmful algal bloom (HAB) species, including Prorocentrum minimum and Heterosigma akashiwo, were observed in bloom concentrations adjacent to the effluent discharge site. Blooms of diatoms were widespread throughout Bishop Harbor. K. brevis was observed with cell concentrations decreasing with increasing proximity to the effluent discharge site. Bioassays using effluent as a nutrient source for K. brevis resulted in decreased cell yields, increased growth rates, and increased time to log-phase growth. The responses of HAB species within Bishop Harbor and of K. brevis to effluent in bioassays suggested that HAB species differ in their response to phosphate-processing effluent.     

Using a macroalgal δ15N bioassay to detect cruise ship waste water effluent inputs

Green macroalgae bioassays were used to determine if the δ¹⁵N signature of cruise ship waste water effluent (CSWWE) could be detected in a small harbor. Opportunistic green macroalgae (Ulva spp.) were collected, cultured under nutrient depleted conditions and characterized with regard to N content and δ¹⁵N. Samples of algae were used in controlled incubations to evaluate the direction of isotope shift from exposure to CSWWE. Algae samples exposed to CSWWE exhibited an increase of 1-2.5‰ in δ¹⁵N values indicating that the CSWWE had an enriched isotope signature. In contrast, algae samples exposed to field conditions exhibited a significant decrease in the observed δ¹⁵N indicating that a light N source was used. Isotopically light, riverine nitrogen derived from N₂-fixing trees in the watershed may be a N source utilized by algae. These experiments indicate that the δ¹⁵N CSWWE signature was not detectable under the CSWWE loading conditions of this experiment.