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.

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.

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.

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.

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.

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.

Hydra, a model system for environmental studies.

Hydra have been extensively used for studying the teratogenic and toxic potential of numerous toxins throughout the years and are more recently growing in popularity to assess the impacts of environmental pollutants. Hydra are an appropriate bioindicator species for use in environmental assessment owing to their easily measurable physical (morphology), biochemical (xenobiotic biotransformation; oxidative stress), behavioural (feeding) and reproductive (sexual and asexual) endpoints. Hydra also possess an unparalleled ability to regenerate, allowing the assessment of teratogenic compounds and the impact of contaminants on stem cells. Importantly, Hydra are ubiquitous throughout freshwater environments and relatively easy to culture making them appropriate for use in small scale bioassay systems. Hydra have been used to assess the environmental impacts of numerous environmental pollutants including metals, organic toxicants (including pharmaceuticals and endocrine disrupting compounds), nanomaterials and industrial and municipal effluents. They have been found to be among the most sensitive animals tested for metals and certain effluents, comparing favourably with more standardised toxicity tests. Despite their lack of use in formalised monitoring programmes, Hydra have been extensively used and are regarded as a model organism in aquatic toxicology.

Comparative toxicity of a brominated flame retardant (tetrabromobisphenol A) on microalgae with single and multi-species bioassays.

The potential threat of emerging chemicals to the aquatic flora is a major issue. The purpose of the study was to develop a multispecies microalgae test in order to determine the impact of species interactions on the cytoxicity of an emergent toxic contaminant: the tetrabromobisphenol A (TBBPA). Single and multi-species tests were thus performed to study the effects of this flame retardant on two microalgae (Pseudokirchneriella subcapitata and Nitzschia palea) commonly observed in freshwater. A synthetic medium was designed to allow the growth of both species. The algae were exposed to 1.8, 4.8, 9.2, 12.9 and 16.5 µM of TBBPA for 72 h. After staining with fluorescein diacetate (FDA), viable cells of each alga species were analyzed by flow cytometry based on chlorophyll autofluorescence and intracellular esterase activity. Density and abundance of viable cells were assessed to follow the population growth and the cell viability. In TBBPA treated samples, the growth of the two microalgae was significantly inhibited at the three highest concentrations (9.2, 12.9 and 16.5 µM) in the two tests. At the end of the experiment (t=72 h), the cell viability was also significantly smaller at these concentrations. The decreases of growth rate and viable cell abundance in TBBPA treated populations of N. palea were significantly higher in multi-species test in comparison with the single-species test. No significant differences were noticed between the two tests for P. subcapitata populations exposed to TBBPA.

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.

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.

An ecotoxicological characterization of nanocrystalline cellulose (NCC).

The pulp and paper industry in Canada is developing technology for the production and use of nanocrystalline cellulose (NCC). A key component of the developmental work is an assessment of potential environmental risks. Towards this goal, NCC samples as well as carboxyl methyl cellulose (CMC), a surrogate of the parent cellulosic material, were subjected to an ecotoxicological evaluation. This involved toxicity tests with rainbow trout hepatocytes and nine aquatic species. The hepatocytes were most sensitive (EC20s between 10 and 200 mg/l) to NCC, although neither NCC nor CMC caused genotoxicity. In tests with the nine species, NCC affected the reproduction of the fathead minnow at (IC25) 0.29 g/l, but no other effects on endpoints such as survival and growth occurred in the other species at concentrations below 1 g/l, which was comparable to CMC. Based on this ecotoxicological characterization, NCC was found to have low toxicity potential and environmental risk.

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.

Toxic effects and bioaccumulation of carbamazepine evaluated by biomarkers measured in organisms of different trophic levels.

The antiepileptic drug carbamazapine (CBZ) readily persists in sewage-water treatment plant wastewaters and finds its way into receiving water bodies. Our study sought to examine the bioaccumulation and toxicity of CBZ using an experimental aquatic trophic chain composed of the green alga, Pseudokirchneriella subcapitata, the crustacean, Thamnocephalus platyurus, and the cnidarian, Hydra attenuata. Bioaccumulation of CBZ was estimated by liquid chromatography-tandem mass spectrometry and revealed bioaccumulation factors of 2.2 and 12.6, respectively, in algae and crustaceans. No significant bioaccumulation was observed in H. attenuata. In T. platyurus, a strong stimulation of global heme oxidase (HO) (76%), and glutathione-S-transferase activity (130%) but a drastic inhibition of cytochrome P450 3A-like activity was found which suggests alteration of enzyme activity by CBZ. However, in the hydranth H. attenuata, an increase in both global cytochrome and cytochrome P450 3A-like activity was found, while GST activity was inhibited. Lipid peroxidation was reduced in T. platyurus and H. attenuata suggesting that redox activity of the lipophilic CBZ was at play. This study highlighted the processes of carbamazepine toxicity transfer between trophic levels in aquatic organisms.

Development of biomarkers of stress related to endocrine disruption in gastropods: alkali-labile phosphates, protein-bound lipids and vitellogenin-like proteins.

The occurrence in aquatic environments of compounds capable of acting as endocrine disruptors (ED) is of concern for wildlife. Elevated levels of the egg-yolk precursor protein vitellogenin (Vg) are widely accepted as a biomarker for xenoestrogen exposure in aquatic organisms. The purpose of this study was to develop a method for tracking changes in levels of Vg-like proteins in gastropods. Vg-like and egg-yolk proteins were analyzed in three freshwater gastropods having different modes of reproduction: Potamopyrgus antipodarum (asexual reproduction), Valvata piscinalis (hermaphroditism) and Lithoglyphus naticoides (sexual reproduction). Vitellogenin-like protein levels were examined by the alkali-labile phosphate (ALP) technique, a novel protein-bound lipid (PBL) assay, and by gradient gel electrophoresis with silver staining. The first phase of the study was dedicated to the development and optimization of an analytical method for detecting Vg-like proteins in these three gastropod species. In the second phase, the snails P. antipodarum and V. piscinalis were exposed to bisphenol A (BPA), octylphenol (OP) and tributyltin (TBT) for 14 and 28 days. Vg-like proteins were resolved in one major band at 250-300 kDa for L. naticoides and V. piscinalis and in two bands at 100 and 30 kDa for P. antipodarum. After 14 days of exposure, all techniques showed an increase in Vg-like protein levels at 100 microg/L BPA and at 1 microg/L OP in P. antipodarum. A decrease in these proteins was observed with high concentrations of OP (100 microg/L) and TBT (>or=5 ng/L). In V. piscinalis, a decrease in Vg-like proteins was shown after 14 days of exposure for OP >or=10 microg/L and TBT >or=5 ng/L; however, at 28 days, gel electrophoresis revealed an increase in these proteins. Histological observations showed significant necrosis in ovotestes of V. piscinalis with the three endocrine-disrupting compounds, while tissue modifications were not detected for P. antipodarum. A method for measuring Vg-like and egg-yolk proteins is proposed to track changes in both estrogenic and androgenic endocrine disruptors in gastropods. The potential use of these species as bioindicators of endocrine disruption in freshwater environments in terms of their reproduction mode is discussed.

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.

The use of biochemical responses to assess ecotoxicological effects of Pharmaceutical and Personal Care Products (PPCPs) after injection in the mussel Elliptio complanata.

A biomarker approach was undertaken using the mussel Elliptio complanata to assess the ecotoxicological effects after injection of a range concentration (0-10mM) of three different PPCPs: carbamazepine, caffeine, methotrexate; and an effluent extract (C8) from St. Lawrence wastewaters treatment plant (Montreal, Canada). A battery of biomarkers, involving oxidative stress and genotoxicity responses: glutation-S-transferase (GST), ethoxyresorufin O-deethylase (EROD), dibenzylflourescein dealkylase (DBF), xanthine oxidoreductase (XOR) activities, lipid peroxidation (LPO) and DNA damage were determined in gonad and digestive gland tissues after 48 h of injection. Results showed an induction of the oxidative metabolism with increasing pharmaceutical concentration in those mussels injected with the PPCPs and the effluent extract. Phase I detoxification enzymes were significantly induced (p<0.05), concretely DBF activity was significantly induced after caffeine, carbamazipine and C8 injection; and EROD activity after C8 and methotrexate injection. Oxidative stress induction only lead to lipid peroxidation (p<0.05) in organisms injected with carbamazepine and caffeine and DNA damage in organisms injected with methotrexate (p<0.05). EROD and DBF enzymatic activities have been found to be suitable biomarkers to determine bioavailability of pharmaceuticals. LPO and DNA damage to determine possible associated adverse effects. Nevertheless, their validation in realistic exposure scenarios and under exposure conditions should be performed in future research.

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.

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.

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.