High concentrations of phthalates affect the early development of the sea urchin Paracentrotus lividus.

The toxicity of three phthalates (PAEs) - butylbenzyl phthalate (BBP), diethyl phthalate (DEP), and di-(2-ethylhexyl) phthalate (DEHP) - was tested on the Mediterranean sea urchin Paracentrotus lividus. Fertilized eggs were exposed to environmental and high PAE concentrations for 72 h. The potential toxic effects on larval development and any morphological anomalies were then assessed to estimate PAEs impact. Environmental concentrations never affected development, while high concentrations induced toxic effects in larvae exposed to BBP (EC50: 2.9 ×103 µg/L) and DEHP (EC50: 3.72 ×103 µg/L). High concentrations caused skeletal anomalies, with a slight to moderate impact for DEP/DEHP and BBP, respectively. PAE toxicity was: BBP>DEHP>DEP. In conclusion, the three PAEs at environmental concentrations do not pose a risk to sea urchins. However, PAE concentrations should be further monitored in order not to constitute a concern to marine species, especially at their early developmental stages.

Nanoplastic uptake temporarily affects the pulsing behavior in ephyrae of the moon jellyfish Aurelia sp.

The aim of this study is to investigate for the first time the uptake and ecotoxicological effects of nanoplastics (NPs) in a marine cnidarian. Ephyrae of the moon jellyfish Aurelia sp. of different ages (0 and 7 days old) were exposed to negatively charged polystyrene NPs for 24 h; then, the uptake was assessed through traditional and novel techniques, namely microscopy and three-dimensional (3D) holotomography. Immobility and behavioral responses (frequency of pulsations) of ephyrae were also investigated to clarify if NP toxicity differed along the first life stages. NP uptake was observed in ephyrae thanks to the 3D technique. Such internalization did not affect survival, but it temporarily impaired the pulsation mode only in 0 day old ephyrae. This may be ascribed to the negative charged NPs, contributing to jellyfish behavioral alteration. These findings promote 3D holotomography as a suitable tool to detect NPs in marine organisms. Moreover, this study recommends the use of cnidarians of different ages to better assess NP ecotoxicological effects in these organisms, key components of the marine food web.

Cold storage effects on lethal and sublethal responses of Amphibalanus amphitrite Nauplii.

Bioassays are extensively used in ecotoxicology and there is a constant need for even more sensitive, reliable and easy to rear and obtain model organisms. Larvae of the crustacean Amphibalanus amphitrite are a good ecotoxicological model, for their high sensitivity to a wide range of toxicants and emerging contaminants. A standardized protocol for this toxicity bioassay has been recently proposed. Nevertheless, a limit of this model organism is the lack of resting stages and the need to use larvae immediately after their release from adults, thus increasing laboratory efforts related to the maintenance of adults. The aim of this work is to verify if short-term cold storage of A. amphitrite larvae prior to use in ecotoxicological tests may affect the ecotoxicological responses of these organisms. Three end-points (mortality, immobilization and swimming speed alteration) were measured on nauplii after storing them at 4 ± 1 °C for different times (24, 72 and 120 h) before bioassay set-up. Bioassays were set up using: (i) clean filtered natural sea water (0.22 µm FNSW), (ii) a reference toxicant (Cadmium Nitrate) and (iii) an environmental matrix (sediment elutriate). Results show that mortality, differently from the other two endpoints, was not affected by cold-storage. Even after 5 days of larvae storage at 4 ± 1 °C before bioassay set up, mortality data were comparable to those obtained for non-cold-stored organisms. Moreover, larval sensitivity to the reference toxicant and sediment elutriate did not change. Regarding the other two end points, low cadmium concentrations significantly changed immobility and swimming activity in cold-stored nauplii compared to larvae used immediately after larval release. In conclusion, short-term cold storage of A. amphitrite nauplii before bioassay set up is an appropriate procedure in ecotoxicological testing if mortality is the endpoint to be considered for final evaluation.

Chemicals sorbed to environmental microplastics are toxic to early life stages of aquatic organisms.

Microplastics are ubiquitous in aquatic ecosystems, but little information is currently available on the dangers and risks to living organisms. In order to assess the ecotoxicity of environmental microplastics (MPs), samples were collected from the beaches of two islands in the Guadeloupe archipelago, Petit-Bourg (PB) located on the main island of Guadeloupe and Marie-Galante (MG) on the second island of the archipelago. These samples have a similar polymer composition with mainly polyethylene (PE) and polypropylene (PP). However, these two samples are very dissimilar with regard to their contamination profile and their toxicity. MPs from MG contain more lead, cadmium and organochlorine compounds while those from PB have higher levels of copper, zinc and hydrocarbons. The leachates of these two samples of MPs induced sublethal effects on the growth of sea urchins and on the pulsation frequency of jellyfish ephyrae but not on the development of zebrafish embryos. The toxic effects are much more marked for samples from the PB site than those from the MG site. This work demonstrates that MPs can contain high levels of potentially bioavailable toxic substances that may represent a significant ecotoxicological risk, particularly for the early life stages of aquatic animals.

Microplastics ingestion in the ephyra stage of Aurelia sp. triggers acute and behavioral responses.

For the first time, we report a correspondence between microplastics (MP) ingestion and ecotoxicological effects in gelatinous zooplankton (Cnidarian jellyfish). The ephyra stage of the jellyfish Aurelia sp. was exposed to both environmental and high concentrations of fluorescent 1-4 µm polyethylene MP (0.01-10 mg/L). After 24 and 48 h, MP accumulation, acute (Immobility) and behavioral (Frequency pulsation) endpoints were investigated. MP were detected by confocal and tomographic investigations on gelatinous body and mouth, either attached on the surface or ingested. This interaction was responsible for impairing ephyrae survival and behavior at all tested concentrations after 24 h. Acute and behavioral effects were also related to mechanical disturbance, caused by MP, triggering a loss of radial symmetry. Contaminated ephyrae exposed to clean seawater showed full recovery after 72 h highlighting the organisms without the microspheres, attached on body jellyfish surface around the mouth and lappets. In conclusion, short-term exposure to MP affects ephyrae jellyfish health, impairing both their survival and behavior. Polyethylene MP temporarily affect both Immobility and Frequency of pulsation of Aurelia sp. jellyfish. This study provides a first step towards understanding and clarifying the potential impacts of MP contamination in gelatinous zooplankton.

A short-term swimming speed alteration test with nauplii of Artemia franciscana.

The presence of toxicant needs to be assessed within short time in order to effectively protect the aquatic environment from serious threat. Based on the observation that at high temperatures aquatic organisms become more vulnerable to stressors than those maintained at room temperature, a new test was developed. The proposed bioassay consisted in the evaluation of the swimming speed alteration (SSA) of nauplii of Artemia franciscana incubated at 39°C (± 1) for 6h, using a Swimming Behavior Recorder system (SBR). A comparative ecotoxicological study between the 6h SSA test and the 24h mortality test was carried out in order to validate the new method in terms of sensitivity by means of EC50 values. The bioassay was applied to screen different toxicants: K2Cr2O7, Cu(SO4)2, NaClO, SDS and Sertraline hydrochloride. The EC50s calculated for the short-term SSA test and those of the mortality test showed comparable values. For all toxicants, the 6h SSA test was proved to be as sensitive as the 24h mortality test. The method developed in this study is the first temperature-based toxicity test with nauplii of Artemia franciscana and it represents an attractive assay in ecotoxicology because of its convenience in terms of time and costs, feasibility and sensitivity.

Effects of polystyrene microbeads in marine planktonic crustaceans.

Plastic debris accumulates in the marine environment, fragmenting into microplastics (MP), causing concern about their potential toxic effects when ingested by marine organisms. The aim of this study was to verify whether 0.1µm polystyrene beads are likely to trigger lethal and sub-lethal responses in marine planktonic crustaceans. MP build-up, mortality, swimming speed alteration and enzyme activity (cholinesterases, catalase) were investigated in the larval stages of Amphibalanus amphitrite barnacle and of Artemia franciscana brine shrimp exposed to a wide range of MP concentrations (from 0.001 to 10mgL-1) for 24 and 48h. The results show that MP were accumulated in crustaceans, without affecting mortality. Swimming activity was significantly altered in crustaceans exposed to high MP concentrations (> 1mgL-1) after 48h. Enzyme activities were significantly affected in all organisms exposed to all the above MP concentrations, indicating that neurotoxic effects and oxidative stress were induced after MP treatment. These findings provide new insight into sub-lethal MP effects on marine crustaceans.

Temperature and salinity effects on cadmium toxicity on lethal and sublethal responses of Amphibalanus amphitrite nauplii.

The official protocol of an ecotoxicological assay employing larvae of the crustacean Amphibalanus amphitrite as a model organism has recently been published by the Italian regulatory authority UNICHIM. Such assay is now one of the applicable tests for water quality assessment under Italian law. While specific temperature and salinity values are recommended by ecotoxicology bioassay protocols for test set up, little information is available on response changes in case of parameter variations. In particular, information is totally lacking for this innovative model organism. Under the standard test protocol, 20°C and 37‰ temperature and salinity, respectively, are required to be set in A. amphitrite bioassay. In order to evaluate the environmental relevance of the test, laboratory experiments simulating the effect on larval responses due to variations of temperature and salinity expected in field collected samples were carried out. The effect of temperature and salinity changes on different end-points, involving increasing sensitivity levels, has been investigated, with and without the presence of cadmium nitrate, Cd(NO3)2, as a reference toxicant, to determine the possible interactions between pollutants and environmental parameters fluctuations. Three end-points - mortality, immobilization, and swimming speed alteration - were measured in order to evaluate the impact of a wide range of temperature (5, 10, 15, 20, 25, 30, 35, 40°C) and salinity values (10, 20, 30, 37, 40, 50, 60, 70‰) on response variation after 24 and 48h of exposure. For each parameter, a Non-Effect Range (NER) - namely the limit values within which no effect related to environmental parameter changes is observed - has been defined. For both parameters, NER resulted to be wider for the less sensitive end-points - such as mortality and immobilization - and for shorter exposure time (24h). Later, the same end-points have been evaluated by exposing the same organisms to a reference toxic compound, Cd(NO3)2 (0, 0.2, 0.4, 0.8, 1.6, 3.2mg/L), within the detected NER both for temperature and salinity. LC50 and EC50 values have been calculated for each end-point after 24 and 48h. Cadmium toxicity was shown to decrease at higher salinity values and increase at higher temperatures. Obtained results offer a better bioassay characterization, and the possibility of a more realistic estimation of ecotoxicological assessments performed on field collected samples. Further studies are needed, especially to investigate the effects of simultaneous salinity and temperature changes on end-points.

Review: Morphofunctional and biochemical markers of stress in sea urchin life stages exposed to engineered nanoparticles.

We describe the use of different life stages of the Mediterranean sea urchin Paracentrotus lividus for the assessment of the possible risk posed by nanoparticles (NPs) in the coastal water. A first screening for the presence of NPs in sea water may be obtained by checking their presence inside tissues of organisms taken from the wild. The ability of NPs to pass from gut to the coelomic fluid is demonstrated by accumulation in sea urchin coelomocytes; the toxicity on sperms can be measured by embryotoxicity markers after sperm exposure, whereas the transfer through the food chain can be observed by developmental anomalies in larvae fed with microalgae exposed to NPs. The most used spermiotoxicity and embryotoxicity tests are described, as well as the biochemical and histochemical analyses of cholinesterase (ChE) activities, which are used to verify toxicity parameters such as inflammation, neurotoxicity, and interference in cell-to-cell communication. Morphological markers of toxicity, in particular skeletal anomalies, are described and classified. In addition, NPs may impair viability of the immune cells of adult specimens. Molecular similarity between echinoderm and human immune cells is shown and discussed. © 2015 Wiley Periodicals, Inc. Environ Toxicol 31: 1552-1562, 2016.

Effects of urea on the molecules involved in the olfactory signal transduction: a preliminary study on Danio rerio.

Among vertebrates, the physiologically uremic Chondrichthyes are the only class which are not presenting the ciliated olfactory receptor neurons in the olfactory neuroepithelium. The only sequenced genome for this class revealed only three olfactory receptor genes and the immunohistochemical detection of G protein alpha subunit typically coupled to the olfactory receptors (Gα(olf)) failed in different species. Chronic renal disease can represent a cause of olfactory impairment in human. In this context, our present study focused on investigating potential effects of high urea concentration on the olfactory epithelium of vertebrates. Larvae of the teleost fish Danio rerio were exposed to urea in order to assess the effects on the olfactory signal transduction; in particular on both the olfactory receptors and the Gα(olf). The endocytosis of neutral red dye in the olfactory mucosa was detected in control and urea-exposed larvae. The amount of neutral red dye uptake was used as a marker of binding and internalization of the Gα(olf). The neutral red dye endocytosis was not affected by urea exposure, hence suggesting that the presence of the Gα(olf) and their binding to the odorants are not affected by urea treatment, either. The presence and distribution of Gα(olf) were investigated in the olfactory epithelium of control and urea-exposed larvae, using a commercial antibody. The immunoreactivity was increased after urea treatment, suggesting an effect of urea on the expression or degradation of this G protein alpha subunit.

Effects of selected metal oxide nanoparticles on Artemia salina larvae: evaluation of mortality and behavioural and biochemical responses.

The aim was to investigate the toxicity of selected metal oxide nanoparticles (MO-NPs) on the brine shrimp Artemia salina, by evaluating mortality and behavioural and biochemical responses. Larvae were exposed to tin(IV) oxide (stannic oxide (SnO2)), cerium(IV) oxide (CeO2) and iron(II, III) oxide (Fe3O4) NPs for 48 h in seawater, with MO-NP suspensions from 0.01 to 1.0 mg/mL. Mortality and behavioural responses (swimming speed alteration) and enzymatic activities of cholinesterase, glutathione-S-transferase and catalase were evaluated. Although the MO-NPs did not induce any mortality of the larvae, they caused changes in behavioural and biochemical responses. Swimming speed significantly decreased in larvae exposed to CeO2 NPs. Cholinesterase and glutathione-S-transferase activities were significantly inhibited in larvae exposed to SnO2 NPs, whereas cholinesterase activity significantly increased after CeO2 NP and Fe3O4 NP exposure. Catalase activity significantly increased in larvae exposed to Fe3O4 NPs. In conclusion, swimming alteration and cholinesterase activity represent valid endpoints for MO-NP exposure, while glutathione-S-transferase and catalase activities appear to be NP-specific.

New insights into the impact of leachates from in-field collected plastics on aquatic invertebrates and vertebrates.

The impact of leachates from micronized beached plastics of the Mediterranean Sea and Atlantic Ocean on coastal marine ecosystems was investigated by using a multidisciplinary approach. Chemical analysis and ecotoxicological tests on phylogenetically distant species were performed on leachates from the following plastic categories: bottles, pellets, hard plastic (HP) containers, fishing nets (FN) and rapido trawling rubber (RTR). The bacteria Alivibrio fischeri, the nauplii of the crustaceans Amphibalanus amphitrite and Acartia tonsa, the rotifer Brachionus plicatilis, the embryos of the sea urchin Paracentrotus lividus, the ephyrae of the jellyfish Aurelia sp. and the larvae of the medaka Oryzias latipes were exposed to different concentrations of leachates to evaluate lethal and sub-lethal effects. Thirty-one additives were identified in the plastic leachates; benzophenone, benzyl butyl phthalate and ethylparaben were present in all leachates. Ecotoxicity of leachates varied among plastic categories and areas, being RTR, HP and FN more toxic than plastic bottles and pellets to several marine invertebrates. The ecotoxicological results based on 13 endpoints were elaborated within a quantitative weight of evidence (WOE) model, providing a synthetic hazard index for each data typology, before their integrations in an environmental risk index. The WOE assigned a moderate and slight hazard to organisms exposed to leachates of FN and HP collected in the Mediterranean Sea respectively, and a moderate hazard to leachates of HP from the Atlantic Ocean. No hazard was found for pellet, bottles and RTR. These findings suggest that an integrated approach based on WOE on a large set of bioassays is recommended to get a more reliable assessment of the ecotoxicity of beached-plastic leachates. In addition, the additives leached from FN and HP should be further investigated to reduce high concentrations and additive types that could impact marine ecosystem health.

Multiple interannual records of young-of-the-year identify an important area for the protection of the shortfin mako, Isurus oxyrinchus.

The shortfin mako (Isurus oxyrinchus) is the second most fishery-exploited pelagic shark in the Mediterranean Sea, thus its conservation status is a cause for concern. Despite the species has been listed in fishery and trade regulations to hinder its population decline, the lack of knowledge on its distribution patterns and habitats essential for its persistence still hampers the implementation of sound conservation actions. Combining data from local expert knowledge, opportunistic catch records, and Baited Remote Underwater Videos, we show evidence of the interannual presence of young-of-the-year (YOY) I. oxyrinchus in the Pelagie Archipelago (Central Mediterranean Sea). A total of twenty-one individuals ranging 71-92.5 cm FL were incidentally caught (on average 2.3 YOY/1000 hooks) or documented on BRUVS in July and August over three consecutive years. These data coupled with questionnaires administered to longline fishers identify one specific area used by YOY in the summer months. Our study presents the most abundant record of YOY shortfin makos in the Mediterranean Sea within such a restricted time and limited area providing important information for improving the protection of this critically endangered species.

Assessing the toxicity of aegerolysin-based bioinsecticidal complexes using the sea urchin Paracentrotus lividus as model organism.

The use of alternative solutions for pest management to replace pesticides in agriculture is of great interest. Proteinaceous complexes deriving from edible oyster mushrooms were recently proposed as environmentally friendly bioinsecticides. Such complexes, composed of ostreolysin A6 (OlyA6) and pleurotolysin B (PlyB), target invertebrate-specific membrane sphingolipids in insect's midgut, causing death through the formation of transmembrane pores. In this work, the potential impact of OlyA6/PlyB complexes was tested in the Mediterranean sea urchin Paracentrotus lividus, as an indicator of environmental quality. The ability of the fluorescently tagged OlyA6 to bind sea urchin gametes (sperm, eggs), the lipidome of sea urchin gametes, and the potential toxic effects and developmental anomalies caused by OlyA6/PlyB complexes on P. lividus early development (embryo, larvae) were investigated. The binding of the fluorescently tagged OlyA6 could be observed only in sea urchin eggs, which harbor OlyA6 sphingolipid membrane receptors, conversely to sperm. High protein concentrations affected sea urchin fertilization (>750 µg/L) and early development (> 375 µg/L in embryos; >100 µg/L in larvae), by causing toxicity and morphological anomalies in embryos and larvae. The main anomalies consisted in delayed embryos and incorrect migration of the primary mesenchyme cells that caused larval skeletal anomalies. The classification of these anomalies indicated a slight environmental impact of OlyA6/PlyB complexes at concentrations higher than 750 µg/L. Such impact should not persist in the marine environment, due to the reversible anomalies observed in sea urchin embryos and larvae that may promote defense strategies. However, before promoting the use of OlyA6/PlyB complexes as bio-pesticides at low concentrations, further studies on other marine coastal species are needed.

Nanomaterial Ecotoxicology in the Terrestrial and Aquatic Environment: A Systematic Review.

This systematic review analyzes the studies available on the ecotoxicity of nanomaterials (NMs) in the environment to understand where future research should be addressed for achieving Agenda 2030 goals on sustainable development and environmental safety. We discuss the status of NMs ecotoxicological effects across different organisms that are representative of all natural environments (land, air, water). A total of 1562 publications were retrieved from the Web of Science (all databases) by using the search criteria "nanomaterials" and "ecotoxicology"; among them, 303 studies were included in the systematic review because they met any of the following criteria: (i) focalize on both search criteria; (ii) deal with terrestrial, or aquatic environment; (iii) address models (organisms, cells) for the nano environmental risk assessment and exposure. The knowledge gaps are identified together with novel insights that need to be further investigated to better understand the ecotoxicological environmental impacts of NMs.

An integrated approach to characterize deep sediment toxicity in Genoa submarine canyons (NW Mediterranean).

The aim of this study was to evaluate deep sediment toxicity in Genoa submarine canyons (Northwestern Mediterranean), for the first time, by using an integrated approach that combined chemistry and ecotoxicology. Sediments were collected from the main submarine canyons in the Gulf of Genoa (Polcevera and Bisagno) and along the adjacent Western Open Slope. A multi-endpoint ecotoxicological approach was taken by exposing two crustacean larvae (Amphibalanus amphitrite and Artemia sp.). Lethal and sub-lethal responses (mortality, swimming behavior) were investigated. Chemical analysis showed that this area is characterized by metal enrichment, including lead, cadmium, chromium, and nickel. Ecotoxicological tests highlighted that elutriates from the different submarine canyons were toxic only for A. amphitrite nauplii: Polcevera Canyon and Western Open Slope sediments induced stronger lethal and sub-lethal ecotoxicological effects than those from Bisagno Canyon. No direct correlation was found between the outcome of chemical and ecotoxicological characterization. However, barnacle was the most prone species to metal contamination: lethal and sub-lethal responses found in this species may be linked to an increase in the concentration of some metals (i.e., Cr, Ni) from offshore to coastal waters, probably due to anthropogenic activity. These findings suggest that the proposed approach can be a suitable tool for deep-sea sediment contamination monitoring; however, the use of a battery of bioassays involving multiple species and endpoints is recommended to better clarify the dynamics of contaminants in marine sediments at very high depths.

Ecotoxicity of Polyvinylidene Difluoride (PVDF) and Polylactic Acid (PLA) Microplastics in Marine Zooplankton.

The aim of this study was to investigate the ecotoxicity of polyvinylidene difluoride (PVDF) and polylactic acid (PLA) microplastics (MPs) in two marine zooplankton: the crustacean Artemia franciscana and the cnidarian Aurelia sp. (common jellyfish). To achieve this goal, (i) MP uptake, (ii) immobility, and (iii) behavior (swimming speed, pulsation mode) of crustacean larval stages and jellyfish ephyrae exposed to MPs concentrations (1, 10, 100 mg/L) were assessed for 24 h. Using traditional and novel techniques, i.e., epifluorescence microscopy and 3D holotomography (HT), PVDF and PLA MPs were found in the digestive systems of the crustaceans and in the gelatinous tissue of jellyfish. Immobility was not affected in either organism, while a significant behavioral alteration in terms of pulsation mode was found in jellyfish after exposure to both PVDF and PLA MPs. Moreover, PLA MPs exposure in jellyfish induced a toxic effect (EC50: 77.43 mg/L) on the behavioral response. This study provides new insights into PLA and PVDF toxicity with the potential for a large impact on the marine ecosystem, since jellyfish play a key role in the marine food chain. However, further investigations incorporating additional species belonging to other trophic levels are paramount to better understand and clarify the impact of such polymers at micro scale in the marine environment. These findings suggest that although PVDF and PLA have been recently proposed as innovative and, in the case of PLA, biodegradable polymers, their effects on marine biota should not be underestimated.

Early-stage anomalies in the sea urchin (Paracentrotus lividus) as bioindicators of multiple stressors in the marine environment: Overview and future perspectives.

The morphological anomalies of the early development stages of the sea urchin Paracentrotus lividus, caused by exposure to environmental stressors, are used as biomarker in ecotoxicological and ecological investigations. Here, we reviewed the available literature and classified the embryo and larval anomalies identified so far, to highlight potential commonalities or differences related to the biological action of the different stressors and their ecological impact. Morphological anomalies are influenced by a) the developmental stage of exposure to stressors; b) the intensity of the stress; c) the intra- and inter-cellular mechanisms affected by the exposure to environmental agents. The classification and analysis of embryo and larvae anomalies, either observed by the authors of this review and reported in literature, indicate that sea urchin abnormalities, caused by exposure to different stressors, can be very similar among them and classified into 18 main types, which can occur individually or mixed. All anomalies can be used to calculate an Index of Contaminant Impact to assess the impact of multiple stressors and to identify relationships between morphological anomalies and compromised biological mechanisms. This approach could be useful for a first screening of the presence of potential stressors impairing the growth and development of the early life stages of marine organisms, thus providing a relevant advancement for in future monitoring activities devoted to assess the health status in coastal marine ecosystems.

The GABAergic-like system in the cyprid of Balanus amphitrite (=Amphibalanus amphitrite) (Cirripedia, Crustacea).

In the present study, biochemical and immunochemical methods were used to investigate the presence and distribution of GABA, glutamate decarboxylase (GAD), GABA(B)R1 and GABA(A) gamma2 subunit receptors and the vesicular GABA transporter (vGAT) in the cyprid of Balanus amphitrite (=Amphibalanus amphitrite). GAD(65/67) immunoreactive neuron cell bodies and nerve fibers were detected in the central nervous system. Paired GAD(65/67) immunoreactive nerves running from the posterior ganglion to the body and limb muscles were detected. Thin GABA-immunoreactive nerve terminals were present on striated muscular fibers and in the antennules. Furthermore, GABA, GAD(65/67), GABA(B)R1 and GABA(A)gamma2 subunit receptors and vGAT were observed in the lateral compound eyes, and GABA(A)gamma2 subunit receptor immunoreactivity was seen in the naupliar eye. These results suggest a neurotransmitter/neuromodulatory role for GABA in thoracic muscle contraction and regulatory functions in compound eyes and antennules of B. amphitrite cyprids.

Swimming speed alteration of Artemia sp. and Brachionus plicatilis as a sub-lethal behavioural end-point for ecotoxicological surveys.

In this study, we investigated the possibility to improve a new behavioural bioassay (Swimming Speed Alteration test-SSA test) using larvae of marine cyst-forming organisms: e.g. the brine shrimp Artemia sp. and the rotifer Brachionus plicatilis. Swimming speed was investigated as a behavioural end-point for application in ecotoxicology studies. A first experiment to analyse the linear swimming speed of the two organisms was performed to verify the applicability of the video-camera tracking system, here referred to as Swimming Behavioural Recorder (SBR). A second experiment was performed, exposing organisms to different toxic compounds (zinc pyrithione, Macrotrol MT-200, and Eserine). Swimming speed alteration was analyzed together with mortality. The results of the first experiment indicate that SBR is a suitable tool to detect linear swimming speed of the two organisms, since the values have been obtained in accordance with other studies using the same organisms (3.05 mm s(-1) for Artemia sp. and 0.62 mm s(-1) for B. plicatilis). Toxicity test results clearly indicate that swimming speed of Artemia sp. and B. plicatilis is a valid behavioural end-point to detect stress at sub-lethal toxic substance concentrations. Indeed, alterations in swimming speed have been detected at toxic compound concentrations as low as less then 0.1-5% of their LC(50) values. In conclusion, the SSA test with B. plicatilis and Artemia sp. can be a good behavioural integrated output for application in marine ecotoxicology and environmental monitoring programs.