Ecotoxicity responses of the freshwater cnidarian Hydra attenuata to 11 rare earth elements.

Lanthanides are the major family of rare earth elements (REEs) owing to the essential properties these metallic species provide in diverse fields of today's world economy. They are now being mined and produced as never before. This raises new environmental concerns in terms of their expected future discharges notably to aquatic systems. Interspecies studies of their ecotoxicity are sparse and effects on aquatic life are still poorly understood. Absence of such information for cnidarians, an ecologically relevant freshwater community, thus prompted the present research on REEs toxicity using Hydra attenuata as our animal model. Lethal and sublethal ecotoxicity data generated with the 11 REEs displayed LC50 values ranging from 0.21 to 0.77 mg L-1and EC50 values ranging from 0.02 to 0.27 mg L-1, thereby confirming the inherent sensitivity of Hydra to REE exposure at environmentally relevant concentrations. Additionally, two properties of REEs were shown to modulate Hydra (sub)lethal toxicity (LC50 and EC50) which decreases with increasing atomic number and with decreasing ionic radius. Compared to studies carried out with different taxonomic groups, Hydra toxicity responses to REEs proved to be among the most sensitive, along with those of other invertebrate species (i.e., Daphnia magna, Ceriodaphnia dubia, Hyalella azteca), suggesting that members of this community are likely more at risk to eventual REE discharges in aquatic environments. Demonstrated Hydra sensitivity to REE exposure strongly justifies their future use in toxicity testing battery approaches to evaluate liquid samples suspected of harbouring REEs.

Effects of a major municipal effluent on the St. Lawrence River: A case study.

The St. Lawrence River (SLR) is the second largest waterway in North America. The discharge of the City of Montreal wastewater treatment plant (WWTP) represents the largest volume of treated wastewaters being released into the river. It also ranks as the largest sewage treatment plant of its kind in North America. Over the last decade, intensive multidisciplinary research has focused on assessing the impacts of Montreal wastewater effluents on the SLR. We describe the major findings of these investigations, including the determination of the fate of contaminants, bioaccumulation in fish and invertebrates, ecotoxicological measurements of aquatic animal health, evaluation of endocrine disruption, parasitism in fish, and combined effects of multiple stressors on the SLR. Impacts of the effluents from the WWTP on aquatic organisms from the SLR are both toxicological and ecological, demonstrating the need for an integrated view of the impacts of municipal effluents on aquatic ecosystems.

Spatial and seasonal variations and ecotoxicological significance of sediment trace metal concentrations in Kebir-Rhumel basin (Northeast of Algeria).

This study sought to assess sediment contamination by trace metals (cadmium, chromium, cobalt, copper, manganese, nickel, lead and zinc), to localize contaminated sites and to identify environmental risk for aquatic organisms in Wadis of Kebir Rhumel basin in the Northeast of Algeria. Water and surficial sediments (0-5 cm) were sampled in winter, spring, summer and autumn from 37 sites along permanent Wadis of the Kebir Rhumel basin. Sediment trace metal contents were measured by Flame Atomic Absorption Spectroscopy. Trace metals median concentrations in sediments followed a decreasing order: Mn > Zn > Pb > Cr > Cu > Ni > Co > Cd. Extreme values (dry weights) of the trace metals are as follows: 0.6-3.4 microg/g for Cd, 10-216 microg/g for Cr, 9-446 microg/g for Cu, 3-20 microg/g for Co, 105-576 microg/g for Mn, 10-46 microg/g for Ni, 11-167 microg/g for Pb, and 38-641 microg/g for Zn. According to world natural concentrations, all sediments collected were considered as contaminated by one or more elements. Comparing measured concentrations with American guidelines (Threshold Effect Level: TEL and Probable Effect Level: PEL) showed that biological effects could be occasionally observed for cadmium, chromium, lead and nickel levels but frequently observed for copper and zinc levels. Sediment quality was shown to be excellent for cobalt and manganese but medium to bad for cadmium, chromium, copper, lead, nickel and zinc regardless of sites.

Ecotoxicology, revisiting its pioneers.

Ecotoxicology is a discipline resulting from pollution events that harmed human and environmental health by the mid-twentieth century. Environmental considerations were simply inexistent at this time, and inevitably deleterious effects and environmental disasters followed. These historical events, like Clear Lake disaster in California, will be recalled, as well as new concepts developed, and scientists involved in these findings. A special tribute is given to Professor Jean-Michel Jouany who conceptualized newly acquired knowledge into an emerging discipline, which he named "ecotoxicology" in the 1960s, and understood to be "toxicology in an ecological perspective." However, René Truhaut is considered as the "father of ecotoxicology" by posterity, while his young mentor Jouany was shadowed by the latter. It is timely to "open the book" as concerns these two exceptional personalities and their working relationships, first to set the record straight and second to give credit where credit is due.

Comparative population genetics of Amblyomma maculatum and Amblyomma americanum in the mid-Atlantic United States.

The Gulf Coast tick, Amblyomma maculatum, is undergoing a northward expansion along the United States East Coast, most recently establishing populations in Virginia, Maryland, and Delaware. This expansion has human health implications, as A. maculatum is the primary natural vector of the bacterium Rickettsia parkeri, which causes a spotted fever-type rickettsiosis. Newly established populations of A. maculatum in Virginia tend to have high prevalence of R. parkeri, compared to lower infection rates in the historical range. The factors contributing to high R. parkeri prevalence in Virginia are not known. Investigating connectivity between sites colonized with A. maculatum can help determine whether sites with higher prevalence are isolated or well-connected through migration, thus serving as a source of infected individuals. We characterized 16S rRNA haplotypes of A. maculatum and, for comparison, the congeneric Amblyomma americanum collected from sites where these species co-occur. We then explored connectivity and genetic structure among Virginia populations using pairwise ΦST and AMOVA analyses. Our study identified one recently restored native grassland site with low A. maculatum haplotype diversity and strong evidence of a founder effect, whereas most sites are haplotypically diverse but with no clear genetic structure or connectivity between sites. These findings contrast with high connectivity and a slight mainland/island structure among A. americanum populations. Our results suggest that A. maculatum populations occasionally arise following long-distance drop-offs of few individual ticks in suitable habitat, but no clear migration patterns were observed. The distinct population genetic patterns between species might result from differences in host utilization.

Phytobenthos and phytoplankton community changes upon exposure to a sunflower oil spill in a South African protected freshwater wetland.

The occurrence of a sunflower oil spill in 2007 in the Con Joubert Bird Sanctuary freshwater wetland, South Africa, inhibited the growth of sensitive phytoplankton species and promoted that of tolerant species. The algal divisions Chlorophyta and Euglenophyta were well represented in the sunflower oil contaminated water, especially the species Euglena sociabilis, Phacus pleuronectes and Chlamydomonas africana. Young and mature resting zygotes of Chlamydomonas africana were recorded in high abundance at all the sunflower oil contaminated sampling sites. The phytobenthos diversity and abundance were significantly suppressed and negatively associated with low Dissolved Oxygen concentrations and the negative redox potential of the bottom sediment. At the intracellular level, phytoplankton chlorophyll a and b concentrations as physiological variables were more sensitive indicators of the adverse effects of sunflower oil than the 72 h Selenastrum capricornutum algal bioassay conducted.

Monitoring of a flame retardant (tetrabromobisphenol A) toxicity on different microalgae assessed by flow cytometry.

Microalgae are key organisms in aquatic ecosystems. Emergent pollutants like the tetrabromobisphenol A (TBBPA) are potential threat for these primary producers at the base of the trophic chain. The effects of this flame retardant on three microalgae (Pseudokirchneriella subcapitata, Nitzschia palea and Chlamydomonas reinhardtii) commonly observed in freshwater ecosystems were studied using a flow cytometer. Each species was exposed to 1.8, 4.8, 9.2, 12.9 and 16.5 µmol L⁻¹ of TBBPA for 72 h. After staining with fluorescein diacetate (FDA), viable cells were discriminated in flow cytogram according to the chlorophyll autofluorescence and the intracellular enzyme activity (esterase) to assess the sensitivity of microalgae to the TBBPA with multi-parametric analysis. For P. subcapitata and N. palea, growth inhibitions of viable cells were lower when the viability was assessed with chlorophyll autofluorescence in comparison with esterase activity. These results are related to the appearance of cells presenting optimal chlorophyll fluorescence without intracellular esterase activity after exposure to TBBPA. Abundance increase of these cells was higher in N. palea than in P. subcapitata. No similar trends were observed in C. reinhardtii populations due to the very high mortality of this microalgal species exposed to TBBPA.

Evaluation of the acute, chronic and teratogenic effects of a mixture of eleven pharmaceuticals on the cnidarian, Hydra attenuata.

Pharmaceuticals have recently emerged as novel pollutants of potential concern in the aquatic environment where they are commonly introduced as complex mixtures via municipal effluent. In the present experiment, the freshwater cnidarian Hydra attenuata was exposed to a mixture of 11 pharmaceuticals (ibuprofen, naproxen, gemfibrozil, bezafibrate, carbamazepine, sulfapyridine, oxytetracycline, novobiocin, trimethoprim, sulfamethoxazole and caffeine) up to 10000 times (x) the concentration found in municipal effluent. Hydra regeneration and teratogenicity was measured, having an IC(50) of 781x and was found to be non teratogenic with an A/D value of approximately 1. Toxicity was investigated using both lethal (based on morphology) and sub-lethal (based on morphology, feeding behaviour, hydranth number and attachment) endpoints. The pharmaceutical mixture incurred a significant decrease in morphology at 0.1, 10 and 100x but a significant increase at 1000x. All parameters were significantly reduced at 10000x. An EC(50) of 425x and 65x based on morphology and feeding respectively and a toxicity threshold (TT) of 3.2x were calculated. When compared to the toxicity of each pharmaceutical exposed individually as previously reported [Quinn B, Gagné F, Blaise C. An investigation into the acute and chronic toxicity of eleven pharmaceuticals found in wastewater effluent on the cnidarian, H. attenuata. Sci Total Environ 2008a; 389: 306-314], the compounds in the mixture were present at concentrations 2 to 3 orders of magnitude lower for the equivalent toxicity (EC(50) and TT). These results indicate that pharmaceuticals act additively in a mixture, having sub-lethal effects at environmentally relevant (microg/L-ng/L) concentrations and that their combined concentrations could potentially prove significantly ecotoxic to Hydra and possibly to other aquatic taxa.

Vitellogenin as a biomarker of exposure to estrogenic compounds in aquatic invertebrates: a review.

Vitellogenins (Vg) are the major precursor of the egg-yolk proteins, vitellins (Vn), which provide energy reserves for embryonic development in oviparous organisms. In mature females, Vg are generally synthesised in response to endogenous estrogens, such as 17beta-estradiol (E2), released into the bloodstream and then stored in developing oocytes. In males, the Vg gene, although present, is normally silent. However, it may be activated by (xeno)-estrogens. These substances belong to a large and heterogeneous group of environmental contaminants capable of altering endocrine functions in organisms. For this reason, they are named endocrine disrupting chemicals (EDCs). Natural steroidal estrogens [E2, estrone (E1), estriol (E3)] and synthetic chemicals, such as pesticides, polychlorinated biphenyls (PCBs) and household products (surfactants) are estrogenic compounds widespread in aquatic ecosystems. The main sources of contamination mostly stem (or originate) from municipal and industrial effluents, agricultural practices, livestock wastes and sewage treatment plants (STP). Recently, Vg induction, mainly in males and immature females, has been proposed as a useful biomarker to assess estrogenic contamination in aquatic environments. Indeed, estrogenicity of individual chemicals and mixtures has extensively been evaluated, in both laboratory and field studies, albeit mostly in fish. In contrast, limited attention has centred on evaluating xenoestrogen effects in aquatic invertebrates, probably owing to limited knowledge of their endocrinology. This review focuses on a brief description of xenoestrogens and their occurrence in aquatic environments, and on Vg induction in aquatic invertebrates in response to both experimental and environmental exposure to estrogenic compounds. Results of recent field and laboratory studies are presented. In addition, new perspectives about the use of Vg induction as a biomarker of exposure to estrogenic compounds are also discussed.

An investigation into the acute and chronic toxicity of eleven pharmaceuticals (and their solvents) found in wastewater effluent on the cnidarian, Hydra attenuata.

Pharmaceuticals previously identified in the effluent from the wastewater treatment plant (WWTP) in Montreal discharging into the St. Lawrence river, were tested for acute and chronic toxicity using the cnidarian Hydra attenuata. Acute toxicity was based on the established technique looking at morphological changes in the Hydra, while recently developed endpoints of feeding behaviour, attachment and growth (hydranth number) were used to measure chronic effects. The compounds under investigation (ibuprofen, naproxen, gemfibrozil, bezafibrate, carbamazepine, sulfamethoxazole, sulfapyridine, oxytetracycline, novobiocin, trimethoprim and caffeine) were tested individually in controlled laboratory exposures with LC(50) and EC(50) results calculated. All compounds tested had relatively high LC(50) values with gemfibrozil, ibuprofen and naproxen having the lowest at 22.36 mg/L and EC(50) values based on morphology of 1.18 to 2.62 mg/L (all concentrations are nominal). The EC(50) values based on feeding were similar to those based on morphology but with increased sensitivity for carbamazepine, bezafibrate and novobiocin. A trend of a reduction in feeding with deterioration in morphology was observed in the Hydra, with the exception of novobiocin, where a lower than expected EC(50) of 13.53 mg/L was found with no negative effect on morphology. Significant reductions in attachment and hydranth number were seen at concentrations of 1 and 5 mg/L for gemfibrozil and ibuprofen respectively. A toxicity threshold (TT) of 320 microg/L was calculated for ibuprofen, only a factor of 10(2) or 10 higher than the concentration found in the effluent in the present study (1.19 mug/L) and in other Canadian effluents studied (22 microg/L [Brun GL, Bernier M, Losier R, Doe K, Jackman P, Lee HB, Pharmaceutically active compounds in Atlantic Canadian sewage treatment plant effluents and receiving waters and potential for environmental effects as measured by acute and chronic aquatic toxicity. Environ Toxicol Chem 2006; 25(8): 2163-2176.] respectively. Using EU directive 93/67/EEC the pharmaceuticals under investigation can be classified as toxic (gemfibrozil, ibuprofen and naproxen), harmful (carbamazepine, bezafibrate, sulfapyridine, oxytetracycline and novobiocin) and non-toxic (sulfamethoxazole, trimethoprim and caffeine) and their potential toxicity for the aquatic environment is discussed.

The effects of pharmaceuticals on the regeneration of the cnidarian, Hydra attenuata.

The Hydra attenuata regeneration assay was used to identify the teratogenic potential of 10 pharmaceuticals identified in effluent from a large city wastewater treatment plant (WWTP). Three types of solvents were used to solubilise the pharmaceuticals (DMSO, acetone and ethanol), at concentrations determined to have no significant effect on measured endpoints. On the one hand, regeneration was significantly inhibited at (nominal) concentrations of 1, 5 and 1 mg/L for gemfibrozil, ibuprofen and naproxen respectively and at the higher concentration of 50 mg/L for bezafibrate and trimethoprim. On the other hand, carbamazepine and the antibiotics sulfapyridine, oxytetracycline and novobiocin significantly increased regeneration at 25, 5, 50 and 50 mg/L respectively. Relatively high IC50 values of 0.9, 3.84, 4.9 and 22.5 mg/L were calculated for gemfibrozil, ibuprofen, naproxen and bezafibrate, respectively. However when subjected to tier two toxicity assessment under EU regulatory guidance using environmentally relevant concentrations a MEC/PNEC value>1 was calculated for gemfibrozil, ibuprofen and naproxen indicating teratogenic potential and the necessity for further tier three assessment. A toxicity index (TI) was also calculated using three different techniques, with TI values>3 (indicating teratogenic potential) found for gemfibrozil, ibuprofen, naproxen and bezafibrate and >1 (indicating a weak teratogenic potential) found for carbamazepine. These results are discussed in the context of their environmental relevance and toxic potential.

Three simple biomarkers useful in conducting water quality assessments with bivalve mollusks.

While biomarkers are undeniably key tools in aquatic ecotoxicology to measure adverse effects linked to contamination events, their application is often inhibited by monetary constraints negating the possibility of having access to dedicated equipment, special wares, and/or expensive reagents. To offset this bottleneck, we propose three simple physiological biomarkers, quantifiable in bivalves, that are free of cost considerations and that can provide basic knowledge on animal health and water quality. Indeed, condition index (CI), growth index (GI), and SOS response (air-time survival) comprise measurements straightforward enough to perform by any laboratory or science body on the planet. Long-term monitoring or screening studies can be carried out with these biomarkers and they are able to provide robust information notably after exposure of bivalves to either singular or multiple agents of contamination. By highlighting examples of data generated in aquatic studies conducted in Eastern Canada under both laboratory and field situations with different species of marine and freshwater mollusks, we establish the suitability of these biomarkers for assessing environmental contamination. Their relationships with other biomarkers are also shown which further corroborate their value as reliable indicators of ecosystem health.

Use of hydra for chronic toxicity assessment of waters intended for human consumption.

Methods developed with the cnidarian, Hydra attenuata (Pallas), have proven effective for screening acute toxicity in aqueous samples, whereas their use in revealing (sub)chronic toxic effects have had mitigated success. We therefore sought to explore manifestations of hydra mortality and abnormal morphological changes, as well as the reproductive capacity of hydras to further enhance the bioassay sensitivity and to assess (sub)chronic toxicity. These parameters were recorded following the onset of experiments after 8, 12 and 19-21 days of hydra exposure. Results obtained with potable waters (30 brands of bottled waters and artesian waters from 9 wells) showed chronic sublethal and lethal effects or reproduction rate inhibition for most samples. The effectiveness of the hydra toxicity test was demonstrated in comparison with other widely used bioassays. Our previous and present investigations suggest that hydra is a reliable and relevant test organism for the assessment of acute and chronic water toxicity.

Exposure of caged mussels to metals in a primary-treated municipal wastewater plume.

The biological availability of metals in municipal wastewater effluents is strongly influenced by the physical and chemical conditions of both the effluent and the receiving water. Aquatic organisms are exposed to both dissolved and particulate (food ingestion) forms of these metals. In the present study, the distribution of metals in specific tissues was used to distinguish between exposure routes (i.e. dissolved vs. particulate phase) and to examine metal bioavailability in mussels exposed to municipal effluents. Caged Elliptio complanata mussels were deployed at sites located between 1.5 km upstream and 12 km downstream of a major effluent outfall in the St. Lawrence River. Metals in surface water samples were fractionated by filtration techniques to determine their dissolved, truly-dissolved (<10 kDa), total-particulate and acid-reactive-particulate forms. At the end of the exposure period (90 days), pooled mussel soft tissues (digestive gland, gills, gonad, foot and mantle) were analyzed for several metals. The results showed that gills and digestive gland were generally the most important target tissues for metal bioaccumulation, while gill/digestive gland metal ratios suggest that both exposure routes should be considered for mussels exposed to municipal effluents. We also found that Ag and Cd in the dispersion plume nearest the outfall, in contrast to other metals such as Cu and Zn, are more closely associated with colloids and were generally less bioavailable than at the reference site in the St. Lawrence River.

4-nonylphenol-induced toxicity and apoptosis in Hydra attenuata.

Effects of 4-nonylphenol (4-NP) on the morphology and survival of the cnidarian Hydra attenuata were studied under acute exposure conditions. The lethal concentration value inducing 50% mortality after 96 h was 97.5 +/- 20 microg/L, whereas the lethal concentration value inducing 10% mortality after 96 h was 64 +/- 25.5 microg/L. The no-observed-effect concentration based on morphological criteria was less than 25 microg/L. Hydra was one of the most sensitive freshwater invertebrate species behind the amphipod Hyalella azteca. Toxicity effects appeared rapidly and did not evolve substantially between 24 and 96 h of exposure. Induction of apoptosis was registered during the first hour of exposure to 4-NP at lethal concentrations, indicating rapid effects of the chemical. Abnormal increase of apoptosis may explain the acute toxicity of 4-NP in hydra. Results show that hydra viability is affected in the short term at 4-NP concentrations normally found in contaminated sites, but not at those concentrations reflecting lower levels of environmental contamination.

Ecotoxicological effects of a semi-submerged municipal dump (Castle harbour, Bermuda) on the Calico scallop Argopecten gibbus.

A biomarker study was undertaken using the Calico scallop Argopecten gibbus to assess the ecotoxicological effects of a semi-submerged municipal dump on the adjacent patch reef lagoon ecosystem (Castle harbour, Bermuda). Caged scallops were deployed in situ for 2 months at various distances from the dump (50 m, 900 m and 2.7 km) and at a reference site (14 km). A suite of biomarkers comprising metallothionein (MT), lipid peroxidation (LPO), vitellin-like proteins (Vn), glutathione S-transferase (GST), DNA strand breaks and condition factor (CF) were investigated in various tissues of the scallop (gill, digestive gland and gonad). Levels of heavy metals were also measured in the whole scallop soft tissue. While there was some variation in response between tissues, in general the results indicated that the dump was negatively impacting scallops deployed in the adjacent marine environment: elevated levels of MT, DNA strand breaks, Vn and GST and reduced condition factor were found for scallops deployed nearest to the dump and at the site 1.5 km from this point source of contamination (Tuckers town) in Castle harbour, with respect to the reference site, North Rock (although this exhibited some degree of metal contamination). The gills from scallops deployed at the dump site were the most responsive tissue, with the highest expression of MT, LPO and DNA damage. This study indicates the potential of the Calico scallop as a convenient bioindicator species in the marine tropical benthos.

Photo-catalysis of bromacil under simulated solar light using Au/TiO2: evaluation of main degradation products and toxicity implications.

Bromacil (5-bromo-3-sec-butyl-6-methyluracil) is a substituted uracil herbicide used worldwide. It is not readily biodegradable and has the potential to contaminate different types of water bodies with possible impact on diverse non-target species. In this work, degradation of bromacil in aqueous Au/TiO2 suspension under simulated sunlight allowed fourteen degradation products to be identified. The photodegradation of bromacil followed (pseudo) first order kinetics in the presence of 0.2 g L(-1) of Au/TiO2 with a half-life of 25.66 ± 1.60 min and a rate constant of 0.0271 ± 0.0023 min(-1). Transformation routes of the photo-catalytic degradation of bromacil were then proposed. Complementary toxicity assessment of the treated bromacil solution revealed a marked decrease in toxicity, thereby confirming that by-products formed would be less harmful from an environmental point of view. Photo-catalytic degradation of bromacil thus appears to hold promise as a cost-effective treatment technology to diminish the presence of this herbicide in aquatic systems.

Microscale Ecotoxicity Testing of Moselle River Watershed (Lorraine Province, France) Sediments.

The ecotoxic potential of seven Moselle river watershed sediments was assessed with a battery of bioassays comprised of rapid phototrophic [LuminoTox solid phase (L-SPA) and elutriate (L-ELU) assays] and bacterial [Microtox solid phase assay (M-SPA)] exposure tests, as well as with two micro-invertebrate solid phase tests conducted with Hydra attenuata (lethal and sublethal effects solid phase assay, HL-SPA and HSL-SPA) and Chironomus riparius. Measured effects of sediments and their elutriates were varied and reflected responses that were ecotoxicity test-, endpoint- and site-dependent, suggesting some degree of risk toward benthic and water column organisms, respectively, at specific sites. Correlation analysis demonstrated that L-SPA and M-SPA ecotoxicity responses were significantly linked with the Hydra HSL-SPA assay, indicating their ability to predict ecotoxicity towards an invertebrate taxonomic group representing secondary consumers. While the L-SPA and M-SPA assays hold promise as rapid screens for sediment ecotoxicity, correlation analysis with grain size (L-SPA: r=-0.795, P=0.033; M-SPA: r=-0.73, P=0.07) points out that their responses can be influenced by the presence of fines (i.e., sediment particles ≤0.063 mm in size) and that this information is essential to properly interpret ecotoxicity data generated with these assays. Finally, notable differences observed in trophic level sensitivities once again recall the importance of employing a test battery to adequately appraise the ecotoxicity of sediments.

Application of rough sets analysis to identify polluted aquatic sites based on a battery of biomarkers: a comparison with classical methods.

The evaluation of toxicological effects at the cellular and molecular levels in organisms are often used to determine sites subjected to contamination problems that pose a threat to the long-term survival of organisms. However, the integration of multiple measurements on the health status of organisms into a model for site discrimination is challenging. This study compares two discrimination methods which are based on rule inference: rough sets (RS) analysis and classification trees (CT) with classical multivariate discriminant analysis (DA). Site classification was attempted with six biomarkers of effects: metallothionein levels, lipid peroxidation, DNA damage, levels of lipophosphoproteins (i.e., vitellins), phagocytosis activity and haemocyte cell viability on clam (Mya arenaria) populations from the Saguenay River fjord (Quebec, Canada). Rule based methods have the advantage of complete independence from data distribution constraints in contrast to the classification methods from multivariate analysis that are more commonly used in ecotoxicology. Results show that RS and CT gave better classifications than DA because they do not require strong distributional assumptions. Moreover, RS provided classification rules that could identify the most important biomarker(s) for site discrimination. RS and CT were shown to be simple and efficient methods for classifying multivariable ecotoxicological data. This methodology would be especially useful when freedom from distributional assumptions is required.

Cyst-based toxicity tests XVI--sensitivity comparison of the solid phase Heterocypris incongruens microbiotest with the Hyalella azteca and Chironomus riparius contact assays on freshwater sediments from Peninsula Harbour (Ontario, Canada).

In view of the complexity and costs of "traditional" whole sediment assays, a "culture/maintenance free" direct contact microbiotest has been developed with the freshwater ostracod Heterocypris incongruens. The new Toxkit assay (named Ostracodtoxkit) has been applied to 33 sediment samples from Peninsula Harbour, located in Lake Superior of the Great Lakes water basin in Ontario, Canada. The microbiotest was applied in parallel to direct contact tests with the amphipod Hyalella azteca and the midge larva Chironomus riparius, to compare its relative sensitivity with that of the two "conventional" assays. The study was undertaken in the framework of remediation action plans for specific areas of concern, to enable decision making by the Canadian authorities for the restoration of impacted aquatic environments. Most sediments were found non-toxic (<20% mortality) to both the conventional test species and the ostracod. For the large majority of samples, a very good correspondence was found between the two crustacean test species for the intensity of the toxic signal "mortality", as reflected by a 0.71 (p<0.05) correlation coefficient. Growth inhibition, which is determined in the ostracod microbiotest as a sublethal effect criterion, allowed the earmarking of some sediment samples, which were apparently more toxic for the amphipod than to the ostracod. For 20% of the samples, substantially higher mortality scores were noted with the ostracod assay than with the midge larvae tests and the overall correlation coefficient between these two tests was lower (r=0.60,p<0.05). The results obtained in the present study corroborate those of previous research on sediments collected from various rivers in Flanders, Belgium, and confirm the potential of the new ostracod microbiotest as a reliable and sensitive low cost alternative for traditional whole sediment assays.