Active Notch signaling is required for arm regeneration in a brittle star.

Cell signaling pathways play key roles in coordinating cellular events in development. The Notch signaling pathway is highly conserved across all multicellular animals and is known to coordinate a multitude of diverse cellular events, including proliferation, differentiation, fate specification, and cell death. Specific functions of the pathway are, however, highly context-dependent and are not well characterized in post-traumatic regeneration. Here, we use a small-molecule inhibitor of the pathway (DAPT) to demonstrate that Notch signaling is required for proper arm regeneration in the brittle star Ophioderma brevispina, a highly regenerative member of the phylum Echinodermata. We also employ a transcriptome-wide gene expression analysis (RNA-seq) to characterize the downstream genes controlled by the Notch pathway in the brittle star regeneration. We demonstrate that arm regeneration involves an extensive cross-talk between the Notch pathway and other cell signaling pathways. In the regrowing arm, Notch regulates the composition of the extracellular matrix, cell migration, proliferation, and apoptosis, as well as components of the innate immune response. We also show for the first time that Notch signaling regulates the activity of several transposable elements. Our data also suggests that one of the possible mechanisms through which Notch sustains its activity in the regenerating tissues is via suppression of Neuralized1.

Kinetics and Equilibrium of Mercury Sorption by Three Different Types of Live Algae

Abstract The kinetics and equilibrium of experimental data of mercury (II) sorption using three different macrophytes E. crassipes, E. azurea and S. ariculata were analyzed. From the kinetic models used, the model 1, which considers the surface area of constant sorption, presents the coefficient of determination, R2, closer to the unit (0.97). Already, in the liquid phase, the best fit of the experimental data was obtained for model 2 (R2=0.96), which considers the variable surface area. The calculated values for the determination coefficients indicate that the Redlich-Peterson isotherm best describes the equilibrium (R2=0.79). The results show that the macrophyte S. ariculata surface area, which presented the highest adsorption potential (15.77x10-4m2.g-1), was far below those found in the best adsorbents. However, considering the large volume of adsorbent material required in an industrial plant and the low cost of the analyzed adsorbents, it is considered that the macrophytes investigated have a considerable potential for the removal of mercury from wastewater.

Characterisation of artisanal mine waste on Buru Island, Indonesia and toxicity to the brittle star Amphipholis squamata.

Artisanal small-scale gold mining (ASGM) using mercury (Hg) amalgamation commenced on Buru Island, Indonesia, in 2012, but was halted in 2015 due to concerns of widespread Hg contamination. Much of the Hg used in the mining process is lost to trommel waste which is disposed of in settlement ponds that drain into adjacent waterways and into Kayeli Bay. Several thousand unmanaged trommel sites and associated tailing ponds exist on Buru Island. This study shows that waste from the Marloso trommel at the Gogrea site contained 203 mg/kg total Hg (THg), with a negligible proportion present as bioavailable methyl Hg (MeHg) and a low total organic carbon content. There are currently very few tools available for ecotoxicological risk assessment of mine tailings for tropical marine ecosystems, and we support the development of Tailings Toxicity Tests (TTTs) and describe laboratory toxicity test methods using the cosmopolitan benthic echinoderm Amphipholis squamata. Undiluted trommel waste caused 100% mortality of A. squamata within 48 h, and a 96-h LC50 of 6.7% w/w trommel waste (4 mg/kg THg) was estimated. Sub-lethal effects on the water vascular system of the brittle star were assessed by quantification of the Ability to Right Itself (ARI), and a 48-h EC50 of 7.3% w/w trommel waste (14.4 mg/kg THg) was estimated. The results show that trommel waste produced on Buru Island is highly contaminated with THg and is acutely toxic, raising serious concern for receiving ecosystems where Hg methylation to more toxic and bioavailable forms is likely.

Regeneration of the digestive system in the crinoid Himerometra robustipinna occurs by transdifferentiation of neurosecretory-like cells.

The structure and regeneration of the digestive system in the crinoid Himerometra robustipinna (Carpenter, 1881) were studied. The gut comprises a spiral tube forming radial lateral processes, which gives it a five-lobed shape. The digestive tube consists of three segments: esophagus, intestine, and rectum. The epithelia of these segments have different cell compositions. Regeneration of the gut after autotomy of the visceral mass progresses very rapidly. Within 6 h after autotomy, an aggregation consisting of amoebocytes, coelomic epithelial cells and juxtaligamental cells (neurosecretory neurons) forms on the inner surface of the skeletal calyx. At 12 h post-autotomy, transdifferentiation of the juxtaligamental cells starts. At 24 h post-autotomy these cells undergo a mesenchymal-epithelial-like transition, resulting in the formation of the luminal epithelium of the gut. Specialization of the intestinal epithelial cells begins on day 2 post-autotomy. At this stage animals acquire the mouth and anal opening. On day 4 post-autotomy the height of both the enterocytes and the visceral mass gradually increases. Proliferation does not play any noticeable role in gut regeneration. The immersion of animals in a 10-7 M solution of colchicine neither stopped formation of the lost structures nor caused accumulation of mitoses in tissues. Weakly EdU-labeled nuclei were observed in the gut only on day 2 post-autotomy and were not detected at later regeneration stages. Single mitotically dividing cells were recorded during the same period. It is concluded that juxtaligamental cells play a major role in gut regeneration in H. robustipinna. The main mechanisms of morphogenesis are cell migration and transdifferentiation.

Functional Morphology of the Arm Spine Joint and Adjacent Structures of the Brittlestar Ophiocomina nigra (Echinodermata: Ophiuroidea).

The skeletal morphology of the arm spine joint of the brittlestar Ophiocomina nigra was examined by scanning electron microscopy and the associated epidermis, connective tissue structures, juxtaligamental system and muscle by optical and transmission electron microscopy. The behaviour of spines in living animals was observed and two experiments were conducted to establish if the spine ligament is mutable collagenous tissue: these determined (1) if animals could detach spines to which plastic tags had been attached and (2) if the extension under constant load of isolated joint preparations was affected by high potassium stimulation. The articulation normally operates as a flexible joint in which the articular surfaces are separated by compliant connective tissue. The articular surfaces comprise a reniform apposition and peg-in-socket mechanical stop, and function primarily to stabilise spines in the erect position. Erect spines can be completely immobilised, which depends on the ligament having mutable tensile properties, as was inferred from the ability of animals to detach tagged spines and the responsiveness of isolated joint preparations to high potassium. The epidermis surrounding the joint has circumferential constrictions that facilitate compression folding and unfolding when the spine is inclined. The interarticular connective tissue is an acellular meshwork of collagen fibril bundles and may serve to reduce frictional forces between the articular surfaces. The ligament consists of parallel bundles of collagen fibrils and 7-14 nm microfibrils. Its passive elastic recoil contributes to the re-erection of inclined spines. The ligament is permeated by cell processes containing large dense-core vesicles, which belong to two types of juxtaligamental cells, one of which is probably peptidergic. The spine muscle consists of obliquely striated myocytes that are linked to the skeleton by extensions of their basement membranes. Muscle contraction may serve mainly to complete the process of spine erection by ensuring close contact between the articular surfaces.

Acute effects of non-weathered and weathered crude oil and dispersant associated with the Deepwater Horizon incident on the development of marine bivalve and echinoderm larvae.

Acute toxicity tests (48-96-h duration) were conducted with larvae of 2 echinoderm species (Strongylocentrotus purpuratus and Dendraster excentricus) and 4 bivalve mollusk species (Crassostrea virginica, Crassostrea gigas, Mytilus galloprovincialis, and Mercenaria mercenaria). Developing larvae were exposed to water-accommodated fractions (WAFs) and chemically enhanced water-accommodated fractions (CEWAFs) of fresh and weathered oils collected from the Gulf of Mexico during the Deepwater Horizon incident. The WAFs (oils alone), CEWAFs (oils plus Corexit 9500A dispersant), and WAFs of Corexit alone were prepared using low-energy mixing. The WAFs of weathered oils had no effect on survival and development of echinoderm and bivalve larvae, whereas WAFs of fresh oils showed adverse effects on larval development. Similar toxicities were observed for weathered oil CEWAFs and WAFs prepared with Corexit alone for oyster (C. gigas and C. virginica) larvae, which were the most sensitive of the tested invertebrate species to Corexit. Mean 10% effective concentration values for total polycyclic aromatic hydrocarbons and dipropylene glycol n-butyl ether (a marker for Corexit) in the present study were higher than all concentrations reported in nearshore field samples collected during and after the Deepwater Horizon incident. The results suggest that water-soluble fractions of weathered oils and Corexit dispersant associated with the Deepwater Horizon incident had limited, if any, acute impacts on nearshore larvae of eastern oysters and clams, as well as other organisms with similar sensitivities to those of test species in the present study; however, exposure to sediments and long-term effects were not evaluated. Environ Toxicol Chem 2016;35:2016-2028. © 2016 SETAC.

Bioindicators as metrics for environmental monitoring of desalination plant discharges.

Development of desalination projects requires simple methodologies and tools for cost-effective and environmentally-sensitive management. Sentinel taxa and biotic indices are easily interpreted in the perspective of environment management. Echinoderms are potential sentinel taxon to gauge the impact produced by brine discharge and the BOPA index is considered an effective tool for monitoring different types of impact. Salinity increase due to desalination brine discharge was evaluated in terms of these two indicators. They reflected the environmental impact and recovery after implementation of a mitigation measure. Echinoderms disappeared at the station closest to the discharge during the years with highest salinity and then recovered their abundance after installation of a diffuser reduced the salinity increase. In the same period, BOPA responded due to the decrease in sensitive amphipods and the increase in tolerant polychaete families when salinities rose. Although salinity changes explained most of the observed variability in both indicators, other abiotic parameters were also significant in explaining this variability.

The effect of copper and temperature on juveniles of the eurybathic brittle star Amphipholis squamata--exploring responses related to motility and the water vascular system.

The limited availability of test organisms that represent tropical and deeper water environments is a significant concern when assessing the risk of contaminants in these environments. Amphipholis squamata (Delle Chiaje 1828) is a widely distributed brittle star with many phylogenetic clades reported from different latitudes, and it also occurs from the intertidal zone to a depth of ∼1300 m. In the present study, the effect of copper on four behavioural responses and mortality of A. squamata were quantified at four different temperatures including 25, 20, 15 and 10°C. At 25°C the four behavioural responses and mortality were relatively sensitive to copper, with 96 h EC50 values of 25 (confidence interval 18-44), 24 (7-26), 32 (24-41), 29 (9-41) µg L(-1) for the measured ability to turn from the oral surface up to oral surface down, curling behaviour, tube foot movement, and tube foot retraction respectively. The average 96-h LC50 value for copper at 25°C was 46 µg L(-1). Some endpoints investigated showed significant effects of reduced temperature compared to the optimal temperature. These effects were enhanced with increasing copper concentrations and significant differences in copper toxicity between temperature treatments were most notable when measuring the ability to turn from the oral surface up to oral surface down where the EC50 changed from 25 (18 to 44) to 6 (-18 to 14) µg L(-1) with a reduction of temperature from 25 to 15°C. The results showed that A. squamata is relatively sensitive to copper and that further investigation into the effects of other stressors on these endpoints is warranted.

Comparative DNA damage and repair in echinoderm coelomocytes exposed to genotoxicants.

The capacity to withstand and repair DNA damage differs among species and plays a role in determining an organism's resistance to genotoxicity, life history, and susceptibility to disease. Environmental stressors that affect organisms at the genetic level are of particular concern in ecotoxicology due to the potential for chronic effects and trans-generational impacts on populations. Echinoderms are valuable organisms to study the relationship between DNA repair and resistance to genotoxic stress due to their history and use as ecotoxicological models, little evidence of senescence, and few reported cases of neoplasia. Coelomocytes (immune cells) have been proposed to serve as sensitive bioindicators of environmental stress and are often used to assess genotoxicity; however, little is known about how coelomocytes from different echinoderm species respond to genotoxic stress. In this study, DNA damage was assessed (by Fast Micromethod) in coelomocytes of four echinoderm species (sea urchins Lytechinus variegatus, Echinometra lucunter lucunter, and Tripneustes ventricosus, and a sea cucumber Isostichopus badionotus) after acute exposure to H2O2 (0-100 mM) and UV-C (0-9999 J/m2), and DNA repair was analyzed over a 24-hour period of recovery. Results show that coelomocytes from all four echinoderm species have the capacity to repair both UV-C and H2O2-induced DNA damage; however, there were differences in repair capacity between species. At 24 hours following exposure to the highest concentration of H2O2 (100 mM) and highest dose of UV-C (9999 J/m2) cell viability remained high (>94.6 ± 1.2%) but DNA repair ranged from 18.2 ± 9.2% to 70.8 ± 16.0% for H2O2 and 8.4 ± 3.2% to 79.8 ± 9.0% for UV-C exposure. Species-specific differences in genotoxic susceptibility and capacity for DNA repair are important to consider when evaluating ecogenotoxicological model organisms and assessing overall impacts of genotoxicants in the environment.

Sewage-exposed marine invertebrates: survival rates and microbiological accumulation.

A large number of bacteria, including agents responsible for diseases, characterise sewage-polluted seawaters. Apart from standards for bathing waters and bivalve aquaculture waters, there are no general microbiological standards applicable to seawaters to help decide if bacterial pollution is within acceptable ranges. This study represents an attempt towards the issue of comparing the susceptibility of different marine invertebrates subjected to polluted seawater with a high microbial contamination. We explored the survival rates and the microbiological accumulation of mollusc bivalves, echinoderms and crustaceans species exposed to sewage-polluted seawaters. Microbiological analyses were performed on the polluted seawater and on the homogenates of exposed and unexposed specimens. Culturable bacteria (22 °C and 37 °C) and microbial pollution indicators (total coliforms, Escherichia coli and intestinal enterococci) were measured. When exposed to the sewage-polluted seawater, the examined invertebrates showed different survival rates. In the filter feeders, bacterial densities at 22 °C and 37 °C rose after 96 h of exposure to sewage. The highest concentrations of total coliforms and intestinal enterococci were found in exposed bivalve Mytilus galloprovincialis. The concentrations of bacteria growing at 37 °C were lower in the exposed deposit feeders compared to the polluted seawater. Some yeasts were absent in several exposed species although these yeasts were present in the polluted seawater. Our data suggest that the examined filter feeders, given their capability to survive and accumulate bacteria, may counteract the effects of sewage and restore seawater quality.

Serotonin and its metabolism in basal deuterostomes: insights from Strongylocentrotus purpuratus and Xenoturbella bocki.

Serotonin (5-HT), an important molecule in metazoans, is involved in a range of biological processes including neurotransmission and neuromodulation. Both its creation and release are tightly regulated, as is its removal. Multiple neurochemical pathways are responsible for the catabolism of 5-HT and are phyla specific; therefore, by elucidating these catabolic pathways we glean greater understanding of the relationships and origins of various transmitter systems. Here, 5-HT catabolic pathways were studied in Strongylocentrotus purpuratus and Xenoturbella bocki, two organisms occupying distinct positions in deuterostomes. The 5-HT-related compounds detected in these organisms were compared with those reported in other phyla. In S. purpuratus, 5-HT-related metabolites include N-acetyl serotonin, gamma-glutamyl-serotonin and 5-hydroxyindole acetic acid; the quantity and type were found to vary based on the specific tissues analyzed. In addition to these compounds, varying levels of tryptamine were also seen. Upon addition of a 5-HT precursor and a monoamine oxidase inhibitor, 5-HT itself was detected. In similar experiments using X. bocki tissues, the 5-HT-related compounds found included 5-HT sulfate, gamma-glutamyl-serotonin and 5-hydroxyindole acetic acid, as well as 5-HT and tryptamine. The sea urchin metabolizes 5-HT in a manner similar to both gastropod mollusks, as evidenced by the detection of gamma-glutamyl-serotonin, and vertebrates, as indicated by the presence of 5-hydroxyindole acetic acid and N-acetyl serotonin. In contrast, 5-HT metabolism in X. bocki appears more similar to common protostome 5-HT catabolic pathways.

Impact of near-future ocean acidification on echinoderms.

As a consequence of increasing atmospheric CO(2), the world's oceans are warming and slowly becoming more acidic (ocean acidification, OA) and profound changes in marine ecosystems are certain. Calcification is one of the primary targets for studies of the impact of CO(2)-driven climate change in the oceans and one of the key marine groups most likely to be impacted by predicted climate change events are the echinoderms. Echinoderms are a vital component of the marine environment with representatives in virtually every ecosystem, where they are often keystone ecosystem engineers. This paper reviews and analyses what is known about the impact of near-future ocean acidification on echinoderms. A global analysis of the literature reveals that echinoderms are surprisingly robust to OA and that important differences in sensitivity to OA are observed between populations and species. However, this is modulated by parameters such as (1) exposure time with rare longer term experiments revealing negative impacts that are hidden in short or midterm ones; (2) bottlenecks in physiological processes and life-cycle such as stage-specific developmental phenomena that may drive the whole species responses; (3) ecological feedback transforming small scale sub lethal effects into important negative effects on fitness. We hypothesize that populations/species naturally exposed to variable environmental pH conditions may be pre-adapted to future OA highlighting the importance to understand and monitor environmental variations in order to be able to to predict sensitivity to future climate changes. More stress ecology research is needed at the frontier between ecotoxicology and ecology, going beyond standardized tests using model species in order to address multiple water quality factors (e.g. pH, temperature, toxicants) and organism health. However, available data allow us to conclude that near-future OA will have negative impact on echinoderm taxa with likely significant consequences at the ecosystem level.

Chemical fate and biological effects of several endocrine disrupters compounds in two echinoderm species.

Two echinoderm species, the sea urchin Paracentrotus lividus and the feather star Antedon mediterranea, were exposed for 28 days to several EDCs: three putative androgenic compounds, triphenyltin (TPT), fenarimol (FEN), methyltestosterone (MET), and two putative antiandrogenic compounds, p,p'-DDE (DDE) and cyproterone acetate (CPA). The exposure nominal concentrations were from 10 to 3000 ng L(-1), depending on the compound. This paper is an attempt to join three different aspects coming from our ecotoxicological tests: (1) the chemical behaviour inside the experimental system; (2) the measured toxicological endpoints; (3) the biochemical responses, to which the measured endpoints may depend. The chemical fate of the different compounds was enquired by a modelling approach throughout the application of the 'Aquarium model'. An estimation of the day-to-day concentration levels in water and biota were obtained together with the amount assumed each day by each animal (uptake in microg animal(-1) d(-1) or ng g-wet weight(-1) d(-1)). The toxicological endpoints investigated deal with the reproductive potential (gonad maturation stage, gonad index and oocyte diameter) and with the regenerative potential (growth and histology). Almost all the compounds exerted some kind of effect at the tested concentrations, however TPT was the most effective in altering both reproductive and regenerative parameters (also at the concentration of few ng L(-1)). The biochemical analyses of testosterone (T) and 17beta-estradiol (E(2)) also showed the ability of the selected compounds to significantly alter endogenous steroid concentrations.

The effects of chronic inorganic and organic phosphate exposure on bactericidal activity of the coelomic fluid of the sea urchin Lytechinus variegatus (Lamarck) (Echinodermata: Echinoidea).

The sea urchin Lytechinus variegatus can survive chronic exposure to sodium phosphate (inorganic phosphate) concentrations as high as 3.2 mg L-1, and triethyl phosphate (organic phosphate) concentrations of 1000 mg L-1. However, chronic exposure to low (0.8 mg L-1 inorganic and 10 mg L-1 organic phosphate), medium (1.6 mg L-1 inorganic and 100 mg L-1 organic phosphate) or high (3.2 mg L-1 inorganic and 1000 mg L-1 organic phosphate) sublethal concentrations of these phosphates inhibit bactericidal clearance of the marine bacterium Vibrio sp. Bacteria were exposed to coelomic fluid collected from individuals maintained in either artificial seawater, or three concentrations of either inorganic phosphate or organic phosphate. Sterile marine broth, natural seawater and cell free coelomic fluid (cfCF) were employed as controls. Bacterial survival indices were measured at 0, 24 and 48 h periods once a week for four weeks. Bacteria were readily eliminated from the whole coelomic fluid (wCF) of individuals maintained in artificial seawater. Individuals maintained in inorganic phosphates were able to clear bacteria following a two week exposure period, while individuals maintained at even low concentrations of organic phosphates failed to clear all bacteria from their coelomic fluid. Exposure to phosphates represses antimicrobial defenses and may ultimately compromise survival of L. variegatus in the nearshore environment.

A dynamic model for predicting chemical concentrations in water and biota during the planning phase of aquatic ecotoxicological tests.

An unsteady-state fugacity model has been developed and validated as a predictive tool that will be useful in the planning phase of aquatic ecotoxicological tests. The model predicts the compound concentration trends in water and biota in experimental aquaria, with respect to the chemical and experimental conditions. The model has been validated with two echinoderm species, Paracentrotus lividus and Antedon mediterranea after a 28-days exposure to p,p'-DDE or triphenyltin chloride (TPT-Cl), respectively. Differences between the predicted vs. measured concentrations of these compounds in water and biota were generally below a factor of two for both compounds. The model here proposed considers three different compartments, water, animals, and glass, and five loss processes: volatilisation, glass adsorption, abiotic degradation, bioconcentration and biotransformation. In particular, adsorption onto glass materials was introduced into the model by means of two equations (R(2) values of 0.86 and 0.90) relating the adsorption rate constant and glass-water partition coefficient on the base of the physical-chemical properties of the compound (log K(ow)). The model can be applied during the planning phase of ecotoxicological tests and for understanding the behaviour of the compound at this micro-ecosystem scale after the tests have been performed.

Echinoderm regenerative response as a sensitive ecotoxicological test for the exposure to endocrine disrupters: effects of p,p'DDE and CPA on crinoid arm regeneration.

Echinoderms are valuable test species in marine ecotoxicology and offer a wide range of biological processes appropriate for this approach. Regenerating echinoderms can be regarded as amenable experimental models for testing the effects of exposure to contaminants, particularly endocrine disrupter compounds (EDCs). As regeneration is a typical developmental process, physiologically regulated by humoral mechanisms, it is highly susceptible to the action of pseudo-hormonal contaminants which appear to be obvious candidates for exerting deleterious actions. In our laboratory experiments, selected EDCs suspected for their antiandrogenic action (p,p'-DDE and cyproterone acetate) were tested at low concentrations on regenerating specimens of the crinoid Antedon mediterranea. An integrated approach which combines exposure experiments and different morphological analyses was employed; the obtained results suggest an overall pattern of plausible endocrine disruption in the exposed samples, showing that processes such as regenerative growth, histogenesis, and differentiation are affected by the exposure to the selected compounds. These results confirm that (1) regenerative phenomena of echinoderms can be considered valuable alternative models to assess the effects of exposure to exogenous substances such as EDCs, and (2) these compounds significantly interfere with fundamental processes of developmental physiology (proliferation, differentiation, etc...) plausibly via endocrine alterations. In terms of future prospects, taking into account the increasing need to propose animal models different from vertebrates, echinoderms represent a group on which ecotoxicological studies should be encouraged and specifically addressed.

Sex steroids and potential mechanisms of non-genomic endocrine disruption in invertebrates.

The review reports on the presence and metabolism of sex steroids in several invertebrate species and provides detailed information on possible mechanisms of endocrine disruption other than the interaction with nuclear receptors. The presence of most vertebrate sex steroids in invertebrate tissues has been demonstrated by liquid or gas chromatography coupled to mass spectrometry. In addition, enzymatic pathways involved in the steroidogenic pathway have been described in at least some invertebrate phyla. Some endocrine disruptors induce alterations in these metabolic pathways and might lead to changes in steroid levels. Growing evidence suggests that estradiol can act through non-genomic pathways in molluscs, and that xenobiotics can as well interfere in these signalling cascades. In spite of these recent advances, most question marks on the action and function of sex steroids in invertebrates remain to be answered.

Endocrine disrupting compounds and echinoderms: new ecotoxicological sentinels for the marine ecosystem.

Echinoderms are valuable test species in marine ecotoxicology and offer a wide range of biological processes appropriate for this approach. In spite of this potential, available data in literature are still rather limited, particularly with regard to the possible effects of endocrine disrupter compounds (EDCs). This review presents echinoderms as useful models for ecotoxicological tests and gives a brief overview of the most significant results obtained in recent years, particularly in the context of the COMPRENDO EU project. In this research project two different aspects of echinoderm physiology, plausibly regulated by humoral mechanisms, were investigated: reproductive biology and regenerative development. Selected EDCs suspected for their androgenic or antiandrogenic action were tested at low concentrations. The results obtained so far showed that different parameters such as regenerative growth, histological pattern, egg diameter and gonad maturation were affected by the exposure to the selected compounds. These results substantiate that reproductive and regenerative phenomena of echinoderms can be considered valuable alternative models for studies on EDCs and confirm that these compounds interfere with fundamental physiological processes, including growth, development and reproductive competence.

Bioavailability of metals along a contamination gradient in San Diego Bay (California, USA).

San Diego Bay is heavily contaminated with metals, but little is known about their biological availability to local marine organisms. This study on 15 elements showed that concentrations of metals associated with sediment increased from the mouth to the back of the Bay while metals in seawater particulates were similar throughout the Bay. Metal bioavailability was assessed over 8weeks following transplant of the local brittlestar, Ophiothrix spiculata (Ophuroidea, Echinodermata), from outside to inside the Bay. Despite a gradient of contamination, brittlestars accumulated similar levels of metals throughout the Bay, suggesting that metal contamination occurred through dissolved metals as well as through the diet. Sediment transplanted in dialysis tubing in the Bay accumulated metals only when placed on the seafloor bottom, indicating greater metal bioavailability near the bottom; the level of accumulation was similar between the mouth and the back of the Bay. The results are consistent with a circulation pattern in which a bottom layer of seawater, enriched with metals, drains from the back to the mouth of the Bay. There was a positive correlation between metal concentration in brittlestars and tidal range, suggesting increased metal exposure due to bay-ocean water exchange. For brittlestar arms the correlation was higher at the mouth than the back of the Bay, indicating greater metal accumulation in arms from dissolved metals in seawater than from ingestion of metal contaminated diet. In contrast, for brittlestar disks the correlation was higher at the back of the Bay, indicative of metal accumulation mainly through the diet. The results highlight the importance of considering bioavailability and physical processes in environmental quality assessments.

Regenerative response and endocrine disrupters in crinoid echinoderms: an old experimental model, a new ecotoxicological test.

The regenerative phenomena that reproduce developmental processes in adult organisms and are regulated by endocrine and neurohumoral mechanisms can provide new sensitive tests for monitoring the effects of exposure to anthropogenic chemicals such as endocrine disrupter (ED) contaminants. These pollutants in fact can be bioaccumulated by the organisms, causing dysfunctions in steroid hormone production/metabolism and activities and inducing dramatic effects on reproductive competence, development and growth in many animals, man included. Current research is exploring the effects of exposure to different classes of compounds well known for their ED activity, such as polychlorinated biphenyls (PCBs), nonylphenols and organotins, on regenerative potential of echinoderms, a relatively unexplored and promising applied approach which offers the unique chance to study physiological developmental processes in adult animals. The selected test species is the crinoid Antedon mediterranea, which represents a valuable experimental model for investigation into the regenerative process from the macroscopic to the molecular level. The present study employs an integrated approach which combines exposure experiments, chemical analysis and biological analysis utilizing classical methods of light (LM) and electron (TEM and SEM) microscopy and immunocytochemistry. The experiments were carried out on experimentally induced arm regenerations in controlled conditions with exposure concentrations comparable to those of moderately polluted coastal zones in order to reproduce common conditions of exposure to environmental contaminants. The results of the exposure tests were analysed in terms of effects at the whole organism, at the tissue and cellular level, and possible sites of action of EDs. Our results show that prolonged exposure to these compounds significantly affects the regenerative mechanisms by inducing appreciable anomalies in terms of regeneration times, overall growth, general morphology and histological and cellular pattern. A concentration/effect relationship could be found for all substances. Interestingly, contrasting results in terms of inhibition or acceleration of regeneration phenomenon were obtained for different chemicals.