Comparative analysis of the jaw apparatus of three marine annelids using scanning electron microscopy: Microstructure and elemental composition.

Polychaeta are highly diversified invertebrates that inhabit marine, brackish or freshwater environments. They have acquired a unique range of adaptative features for securing food. However, the jaw apparatus may reveal not only defence and predation mechanisms, but also its relation to environmental chemistry. The present work compared the structure and chemical profile of the jaws of different estuarine Polychaeta: Nephtys hombergii (Nephtyidae), Hediste diversicolor (Nereididae) and Glycera alba (Glyceridae) using Scanning Electron Microscopy (SEM) and Scanning Electron Microscopy with Energy Dispersive X-Ray (SEM-EDX). Analyses revealed that N. hombergii possesses a muscular jawless proboscis with terminal sensorial papillae for detecting prey, whereas the G. alba proboscis exhibits four delicately sharp jaws with perforations for venom delivery and H. diversicolor bears two blunt denticulated jaws to grasp a wide variety of food items. Melanin and metals like copper provide hardness to the slender jaws of Glycera, while, in the absence of heavier metallic elements, halogens contribute to H. diversicolor jaws robustness. The more specific chemistry of the jaws of glycerids is associated with its more refined venom injection, whereas Hediste is an opportunistic omnivore and Nepthys an agile forager. Altogether, the chemistry of jaws is an adaptive feature for feeding, locomotion and even resilience to complex and often adverse chemical profiles of estuaries.

Calcium homeostasis and stable fatty acid composition underpin heatwave tolerance of the keystone polychaete Hediste diversicolor.

Extreme weather events, such as heatwaves, are becoming increasingly frequent, long-lasting and severe as global climate change continues, shaping marine biodiversity patterns worldwide. Increased risk of overheating and mortality across major taxa have been recurrently observed, jeopardizing the sustainability of ecosystem services. Molecular responses of species, which scale up to physiological and population responses, are determinant processes that modulate species sensitivity or tolerance to extreme weather events. Here, by integrating proteomic, fatty acid profiling and physiological approaches, we show that the tolerance of the intertidal ragworm Hediste diversicolor, a keystone species in estuarine ecosystems and an emergent blue bio-resource, to long-lasting heatwaves (24  vs 30 °C for 30 days) is shaped by calcium homeostasis, immune function and stability of fatty acid profiles. These features potentially enabled H. diversicolor to increase its thermal tolerance limit by 0.81 °C under the heatwave scenario and maintain survival. No growth trade-offs were detected, as wet weight remained stable across conditions. Biological variation of physiological parameters was lower when compared to molecular measures. Proteins showed an overall elevated coefficient of variation, although decreasing molecular variance under the heatwave scenario was observed for both proteins and fatty acids. This finding is consistent with the phenomenon of physiological canalization in extreme environments and contradicts the theory that novel conditions increase trait variation. Our results show that keystone highly valued marine polychaetes are tolerant to heatwaves, confirming the potential of H. diversicolor as a blue bio-resource and opening new avenues for sustainable marine aquaculture development.

Toxicological evaluation of sediment samples spiked with human pharmaceutical products: Energy status and neuroendocrine effects in marine polychaetes Hediste diversicolor.

There is a lack of studies about the ecotoxicology of pharmaceutical products on marine environment. To predict possible adverse effects of pharmaceutical products on benthic biota, polychaetes Hediste diversicolor were exposed for 14-days to pharmaceutical-spiked sediments under laboratory conditions. Carbamazepine (CBZ), ibuprofen (IBP) and propranolol (PRO) at concentrations of 500ngg(-1), 50ngg(-1), 5ngg(-1), 0.5ngg(-1) and 0.05ngg(-1), fluoxetine (FX) and 17α-ethynylestradiol (EE2) at concentrations of 100ngg(-1), 10ngg(-1), 1ngg(-1), 0.1ngg(-1) and 0.01ngg(-1), including environmental concentrations (underlined), were spiked in marine sediment samples. After the exposure, cellular energy status (total lipids content - TLP; and mitochondrial electron transport activity - MET), metabolism of monoamines (monoamine oxidase activity - MAO) and inflammation properties (cyclooxygenase activity - COX) were observed in polychaetes. CBZ increased TLP content and MET activity, and decreased MAO activity in polychaetes. IBP did not interfere on the TLP level, but on the MET and MAO activities (environmental concentrations). FX did not cause changes in the energy status. Therefore, environmental concentration diminished MAO activity. EE2 did not affect the energy status, however, MAO activity was significantly lower in polychaetes exposed to environmental concentration. PRO increased TLP level in polychaetes, but not MET activity. MAO activity was significantly lower for polychaetes exposed to environmental concentration. Except FX, all pharmaceuticals showed anti-inflammatory properties confirmed by the decrease of COX activity. Pharmaceutical products affected H. diversicolor physiology and health. As a benthic top predator, adverse effects on sea-worms can potentially culminate in ecosystem perturbations.

A whole-body transcriptome assembly of the annelid worm Hediste diversicolor.

The Annelida phylum is composed of a myriad of species exhibiting key phenotypic adaptations. They occupy key ecological niches in a variety of marine, freshwater and terrestrial ecosystems. Importantly, the increment of omic resources is rapidly modifying the taxonomic landscape and knowledge of species belonging to this phylum. Here, we comprehensively characterised and annotated a transcriptome of the common ragworm, Hediste diversicolor (OF Müller). This species belongs to the family Nereididae and inhabits estuarine and lagoon areas on the Atlantic coasts of Europe and North America. Ecologically, H. diversicolor plays an important role in benthic food webs. Given its commercial value, H. diversicolor is a promising candidate for aquaculture development and production in farming facilities, under a circular economy framework. We used Illumina next-generation sequencing technology, to produce a total of 105 million (M) paired-end (PE) raw reads and generate the first whole-body transcriptome assembly of H. diversicolor species. This high-quality transcriptome contains 69,335 transcripts with an N50 transcript length of 2313 bp and achieved a BUSCO gene completeness of 97.7% and 96% in Eukaryota and Metazoa lineage-specific profile libraries. Our findings offer a valuable resource for multiple biological applications using this species.

Towards the understanding of the uptake and depuration of microplastics in the ragworm Hediste diversicolor: Field and laboratory study.

An important number of studies have evaluated the presence of microplastics, particles with a size below 5 mm, in aquatic organisms. Studies have shown that these fragments are widely present in the marine environment, but research on the estuarine ecosystem is still scarce. In this study, two different approaches were used to evaluate the presence and ingestion of plastic particles in the ragworm Hediste diversicolor: a field study for the environmental assessment and a laboratory experiment in controlled condition. For the environmental evaluation, ingestion of microplastics was evaluated in the ragworm H. diversicolor sampled from the mudflats of the Seine estuary (France) during March and June 2017 and 2018, on two locations: S1 and S2, both characterized by high anthropogenic pressures, and for S2 a more influential hydrodynamic component. Ingestion of microplastics was measured in ragworms tissues and in gut content (sediment) after depuration. The number of particles as well as their size, shape and color were reported and compared between sampling period and locations. Results showed the presence of a low number of particles in both worms and gut content. In gut content, 45.6% and 87.58% of samples from site S1 and S2 respectively contained plastic like particles. In worms, 41.7% (S1) and 75.8% (S2) of analysed samples contained plastic like items. The lowest mean number of particles was 0.21 ± 0.31 (S1 in June 2017) in worms' tissues, but 0.80 ± 0.90 (S1 in June 2017) in the gut content and the highest was 1.47 ± 1.41 (S2 in April 2017) while the highest number was 2.55 ± 2.06 (S2 in June 2017) in worms and gut content respectively. The majority of suspected microplastics observed were fibers (66%) and fragments (27%), but films (3.7%) foam (2.1%), and granules (0.2%) were also identified. In addition, the most polymer type observed by Raman spectroscopy was polypropylene. Furthermore, a preliminary study of the ingestion and egestion of fluorescent polyethylene (PE) microbeads in the digestive tract of ragworms was conducted after exposure through water, during 1h at 1.2 × 106 MP/mL. Results showed a rapid turnover of PE microbeads throughout the digestive tract of worms especially after exposure through water. This study revealed that microplastics are ingested by the ragworm H. diversicolor but do not seem to bioaccumulate. More research is needed to measure potential chronic effects of microplastics on physiological parameters of H. diversicolor and potential trophic transfer of microplastics.

Effect of an environmental microplastic mixture from the Seine River and one of the main associated plasticizers, dibutylphthalate, on the sentinel species Hediste diversicolor.

The aim of this study was to explore the adverse effects of a microplastic (MP) mixture obtained from litter accumulated in the Seine River (France) compared to those of their major co-plasticizer, dibutylphthalate (DBP), on the sentinel species Hediste diversicolor. A suite of biomarkers has been investigated to study the impacts of MPs (100 mg kg-1 sediment), DBP (38 µg kg-1 sediment) on worms compared to non-exposed individuals after 4 and 21 days. The antioxidant response, immunity, neurotoxicity and energy and respiratory metabolism were investigated using biomarkers. After 21 days, worms exposed to MPs showed an increasing aerobic metabolism, an enhancement of both antioxidant and neuroimmune responses. Energy-related biomarkers demonstrated that the energy reallocated to the defence system may come from proteins. A similar impact was depicted after DBP exposure, except for neurotoxicity. Our results provide a better understanding of the ecotoxicological effects of environmental MPs and their associated-contaminants on H. diversicolor.

Innovative in vivo and in vitro bioassays for the establishment of toxicity thresholds of pollutants in sediment quality assessment using polychaetes and their immune cells.

Sediment toxicity testing has become a crucial component for assessing the risks posed by contaminated sediments and for the development of sediment quality assessment strategies. Commonly used organisms for bioassays with estuarine sediments include amphipods, Arenicola marina polychaetes and echinoids. Among the latter, the Sea Urchin Embryo test (SET) is the most widely used. However, one relevant limitation of this bioassay is the unavailability of gametes all year-round, particularly outside the natural spawning seasons. Consequently, the establishment of an appropriate and complementary model organism for a continuous assessment of sediment quality is recommended. A reliable assessment of the hazards resulting from pollutants in sediments or pore water, can be achieved with ecologically relevant species of sediment such as the polychaete Hediste diversicolor, which is widespread in estuaries and has the capacity to accumulate pollutants. The aim of this work was to develop reliable in vivo and in vitro bioassays with H. diversicolor and its coelomocytes (immune cells) to determine the toxicity thresholds of different contaminants bounded to sediments or resuspended into water. Polychaetes were exposed to sublethal concentrations of CuCl2 (in vivo) and a non-invasive method for collection of polychaetes coelomocytes was applied for the in vitro bioassay, exposing cells to a series of CuCl2 and AgNPs concentrations. Same reference toxicants were used to expose Paracentrotus lividus following the SET (ICES Nº 51; Beiras et al., 2012) and obtained toxicity thresholds were compared between the two species. In vivo exposure of polychaetes to high concentrations of Cu produced weight loss and histopathological alterations. After in vitro approaches, a significant decrease in coelomocytes viability was recorded for both toxicants, in a monotonic dose-response curve, at very short-exposure times (2 h). The toxicity thresholds obtained with polychaetes were in line with the ones obtained with the SET, concluding that their sensitivity is similar. In conclusion, in vivo and in vitro bioassays developed with H. diversicolor are accurate toxicity screenings of pollutants that could be bounded to sediments or dissolved in the pore water, and may complement the SET outside the spawning period of the echinoderms. The bioassays herein developed could be applied not only to establish the toxicity thresholds of individual compounds or mixtures, but also to assess the toxicity of field collected sediments.

Time-course distribution of fluorescent microplastics in target tissues of mussels and polychaetes.

The majority of the plastic produced in the last century is accumulated in the environment, leading to an exacerbated contamination of marine environments due to transport from land to the ocean. In the ocean, mechanical abrasion, oxidation, and photodegradation degrade large plastics into microplastics (MPs) - 0.1 µm to 5 mm (EFSA, 2016) which are transported through water currents reaching the water surface, water column, and sediments. Further, they can be accumulated by aquatic and benthic species, entering the trophic chain and becoming a potential threat to humans. In the present research, we aimed to decipher the accumulation and distribution time-courses between different organs or target tissues of organisms inhabiting coastal areas such as mussels Mytilus galloprovincialis and polychaetes Hediste diversicolor. Both were exposed in microcosm experiments to fluorescent polystyrene MPs (1 µm) which were spiked at two doses (103 and 105 particles/mL) for 1, 4, 24, and 72 h. Mussels and polychaetes were digested with 10% KOH and filtered to quantify the number of MPs incorporated. Different anatomical parts of the body were selected and processed for cryosectioning and posterior microscopic localisation of MPs. Both species accumulate MPs spiked in water column, mainly after exposure to the highest dose. In mussels, particles were found in distinct parts of the digestive tract (stomach, digestive diverticula, ducts) and gills. Even if the majority of MPs were localised in the lumen of the digestive tract, in some cases, were inside the digestive epithelium. The identification of MPs and their internalization in the digestive system was studied using Raman spectroscopy. A decreasing trend with time regarding MPs number in the digestive tract (stomach) of mussels was observed while the opposite was recorded for polychaetes and sediments. The combination of microscopical observations of frozen sections and Raman, appeared to be accurate methodologies to address MPs abundances and to reveal their localisation in different organs. This work has enabled to understand the distribution and fate of MPs in different environmental compartments and it could contribute to gain knowledge about their impact after ingestion by coastal organisms.

An investigation into the toxicity of tissue extracts from two distinct marine Polychaeta.

The present study investigated the potential toxicity of venomous secretions of two polychaetes, Hediste diversicolor and Glycera alba (Annelida: Phyllodocida). Toxic activity of putative toxins, measured on mussel gills through the Comet assay, revealed higher effects caused by extracts from H. diversicolor skin and G. alba specialised, jawed proboscis, when compared to control. The results suggest that H. diversicolor secretes toxins via skin for protection against predators, contrarily to G. alba, who secretes toxins for predation.

Investigating the impact of remediation efforts on Hedistediversicolor in the Seine estuary using multiple levels of biological organization.

The Seine Estuary is historically one of the most contaminated estuaries in Europe. In 2002 the estuary underwent major redevelopment which emphasized environmental remediation to measure biological effects. The present study investigated the health status of the sentinel species Hediste diversicolor in the Seine estuary by comparing data from a sampling campaign immediately after the Seine underwent reconstruction and 10 years afterward. Both studies implemented multiple levels of biological organization ranging from enzymatic biomarkers to population density. Integrative modeling was used to establish a holistic status assessment for H. diversicolor in the Seine. H. diversicolor populations in the Seine estuary were impacted in both campaigns suggesting no significant improvements over the 10-year period. This interpretation however can be improved by increasing the number of reference sites used to establish the baseline natural variation, integrating additional environmental parameters into the model and measuring additional biological endpoints, such as reproductive status.

Are tolerance processes limiting the responses of Hediste diversicolor to cadmium exposure? A multimarker approach.

Cadmium (Cd) is considered a priority hazardous substance under the European Community Directive 2013/39 due to its ecotoxicity. The ragworm Hediste diversicolor (O.F. Müller, 1776), a common species in estuaries and coastal lagoons, plays an important ecological role in these ecosystems and is a suitable bioindicator of environmental chemical contamination. In this study, H. diversicolor was chosen as an ecotoxicological model with the aim of evaluating the responses to Cd contamination, considering a multi-biomarker approach (mortality, biometry, behaviour, Cd bioaccumulation, oxidative stress and damage, and energy metabolism). Also, the hypothesis of different tolerances resulting in different responses was evaluated, by collecting worms from three systems distinctly impacted by metal contamination (Mondego estuary, Óbidos Lagoon and Sado estuary - Portugal). Animals were exposed under laboratory conditions to cadmium (10, 50 and 100 µg/L), for 10 days. Significant differences were observed in responses amongst worms originating from the different sites. Organisms from the less impacted systems revealed greater effects on mortality, biomass decrease and burrowing behaviour, as well as higher bioaccumulation potential, after exposure to Cd. Biochemical and behaviour impairments were observed as a consequence of Cd exposure, although not in a concentration-dependant manner. The results obtained in this study reinforce the importance of integrating endpoint responses, at the individual and sub-individual levels, to assess potential changes induced by pollutants in the physiological status and fitness of H. diversicolor and help to predict what their ecological consequences might be.

Uptake, accumulation and associated cellular alterations of environmental samples of microplastics in the seaworm Hediste diversicolor.

The ubiquitous distribution of microplastics (MPs) in the marine environment raises global concern to understand their impact. Environmental MPs have been shown to exhibit different physicochemical properties during their life cycles. However, the body of knowledge regarding their accumulation and biological effects is still significantly limited compared to manufactured MPs. To evaluate the hazardous effects of a mixture of environmental MPs collected along the Tunisian beaches, their accumulation and cellular effects were investigated in Hediste diversicolor. MP sample was composed of polyethylene (PE), polyethylene vinyl acetate (PEVA), low-density polyethylene (LDPE), high-density polyethylene (HDPE), polypropylene (PP) and polyamide (PA) analyzed using Raman microspectroscopy (RM). The concentrations of MPs in seaworm tissues increased over time, following the order 1.2-0.45 µm > 3-1.2 µm > 100-3 µm. The ingestion of MPs by H. diversicolor reduced their survival and growth, affected the neuro-transmission and antioxidant pathways. Our data emphasised that the toxic effects of environmental MPs were closely related to the exposure dose and period. The results also demonstrated that the size distribution of MPs in seaworms was mainly correlated with biochemical markers. This study highlights the ecological risk in the ingestion and accumulation of environmental MPs by biota that threatens their functional parameters.

Experimental evidence of uncertain future of the keystone ragworm Hediste diversicolor (O.F. Müller, 1776) under climate change conditions.

It is currently assumed that climate change related factors pose severe challenges to biodiversity maintenance. This paper assesses the multi-stressor effects of elevated temperature (15 °C as control, 25 °C as elevated) and CO2 levels (pH 8.1 as control, 7.5 and 7.0 representing acidifying conditions) on the physiological (survival and regenerative capacity), behavioral (feeding and burrowing activities), and biochemical changes (metabolic capacity, oxidative status and biotransformation mechanisms) experienced by the keystone polychaete Hediste diversicolor. Temperature rise enlarged the adverse effect of marine acidification on the survival of H. diversicolor, delayed the beginning of the excavation activity, enhancing the negative effects that pH decrease had in the burrowing behavior of this polychaete. Additionally, regardless of the temperature, exposure of H. diversicolor to acidification results in a reduction in the feeding rate. It is the first time that this decreased feeding capacity is found related to seawater acidification in this species. The healing of the wound and the blastemal formation were retarded due to these two climatic factors which hinder the regenerative process of polychaetes. These vital physiological functions of H. diversicolor can be related to the oxidative stress induced by climate change conditions since free radicals overproduced will impair cells functioning, affecting species biochemical and physiological performance, including feeding and tissue regeneration. The present results also demonstrated that although polychaetes' metabolic capacity was enhanced under stress conditions, organisms were still able to increase or maintain their energy reserves. Our findings are of major environmental relevance considering that predicted climate change conditions will affect species vital and ecological and physiological capacities. These can be translated into shrinking not only at the individual and population level but also in microbial and endofaunal diversities, in the detritus processing in estuaries and biogeochemical cycles at the ecosystem level. Thus the conservation of H. diversicolor populations is vital for the normal functioning of estuarine mudflat ecosystems.

Complete mitochondrial genome of the marine polychaete Hediste japonica (Phyllodocida, Nereididae).

To date, only five species are registered in the genus Hediste, and complete mitochondrial genome is reported in one species, Hediste diadroma. In this study, a complete 15,783 bp genome for the marine polychaete H. japonica mitochondrion was assembled through Illumina HiSeq platform. The complete mitochondrial genome of H. japonica contained 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, two ribosomal RNA (rRNA) genes, and one control region. Overall genomic structure and gene orientation of H. japonica mitogenome are identical to those of H. diadroma. Phylogenetic analysis using the maximum likelihood method validated the sister relationship between Hediste sp. and other polychaetes. This information will be useful to understand geographical distribution, phylogenetic relationship, and evolutionary history of marine polychates.

First characterization of the ecotoxicity of paraffin particles: assessment of biochemical effects in the marine polychaete Hediste diversicolor.

Plastics are a widespread environmental problem, particularly in the form of small particles or fragments (microplastics). With the purpose of gradually replacing the use of plastics in cosmetic products (primary microplastics), the use of paraffin in microspheres has recently been applied. Paraffin waxes are composed by organic molecules usually derived from petroleum, thus assumed to be chemically and biologically inert, having a lower (eco)toxicity and residence time compared with plastic particles. However, the low ecotoxicity of paraffin waxes may be somewhat questionable, because some paraffin constituents can be absorbed and catabolized, thus exerting biological effects. The main objective of this study was to understand the potential toxic effects of paraffin microparticles on key physiological processes of the polychaete Hediste diversicolor. To attain this objective, this work assessed the acute and chronic effects of three densities (5, 20, and 80 mg L-1) of four size ranges of paraffin particles (from 100 to 1200 µm) on this epibenthic organism. Although paraffin wax can be generically considered innocuous, the present study was able to demonstrate a significant inhibition in the activity of acetylcholinesterase in acutely exposed organisms and an increase in the activity of glutathione peroxidase, catalase, and glutathione S-transferase under some specific scenarios.

Accumulation and immunotoxicity of microplastics in the estuarine worm Hediste diversicolor in environmentally relevant conditions of exposure.

The presence of plastic debris < 5 mm called microplastics (MPs) which results mainly from macroplastic's fragmentation has been reported in aquatic ecosystems. Several studies have shown that MPs are persistent and their accumulation was observed in various aquatic species. However, the majority of studies focused on marine species, and much less on continental and estuarine biota. The goal of the present study was to investigate the effects of a mixture of two types of MPs (polyethylene and polypropylene), frequently found in natural environments, towards the ragworm Hediste diversicolor to determine their accumulation in organisms exposed through the water phase or sediment. Two concentrations of exposure were selected for medium and heavily contaminated areas reported for water phase (10 and 100 µg/L) and sediment (10 and 50 mg of MPs/kg). To study the potential toxic effect of MPs, immune parameters were selected since they are involved in many defense mechanisms against xenobiotics or infectious agents. An average number of MP items/worm ranging from 0 to 2.5 and from 1 to 36 were identified in animals exposed to the lowest and the highest concentration of MPs through water exposure. In worms exposed through sediment, less than 1 MP/worm was found and a greater number of particles were identified in depurated sediment. For immunotoxic impact, MP exposure induced a decrease in coelomocytes viability, but no alteration of phagocytosis activity, phenoloxydase, and acid phosphatase was measured. This study brings new results on the potential accumulation and immunotoxicity of MPs for the ragworm H. diversicolor who plays a key role in the structure and functioning of estuarine ecosystem.

Study of the effects of zinc pyrithione in biochemical parameters of the Polychaeta Hediste diversicolor: evidences of neurotoxicity at ecologically relevant concentrations.

Nowadays there are various groups of biocidal chemical agents, which can be used in diverse areas, such as personal hygiene, disinfection, antiparasitic action, and also in antifouling mixtures or paints. The versatility and efficacy of some of these agents favors their use and ultimate release into the aquatic environment, where they may still exert toxic activity. Zinc pyrithione is classified as a metal biocide with bactericidal, algicidal, and fungicidal actions. It has been formulated in antifouling paints, which prevent the formation of biofilms in submerged structures, and has also been used for dermocosmetic purposes, in shampoos for the treatment of dandruff and seborrhea. Some of the uses of zinc pyrithione are responsible for its direct release as flakes that reach the bottom sediments, especially in estuarine areas. Considering this fate, the ecotoxicity assessment of its effects towards sediment organisms, namely Polychaetous species, is extremely important. The present study characterized the acute potential toxicity of zinc pyrithione in terms of parameters of oxidative stress (catalase, glutathione S-transferases (GSTs), and thiobarbituric acid reactive substances (TBARS)), and neurotoxicity (acetylcholinesterase) which were evaluated in individuals of the polychaete Hediste diversicolor. Regarding the results obtained, only the activity of GSTs and AChE was significantly altered in relation to non-exposed animals. This set of results indicates that oxidative stress did not occur.

Is there a link between acetylcholinesterase, behaviour and density populations of the ragworm Hediste diversicolor?

The main objective of the present study was to explore the potential link between acetylcholinesterase (AChE) activity and burrowing behaviour of the ragworm Hediste diversicolor, which may have consequences at higher levels of biological organisation. Two complementary studies were conducted. AChE activity, at the sub-individual level, and behavioural responses, at the individual level, were evaluated in worms from the Loire estuary (France), whereas density and biomass of H. diversicolor were determined at the population level. A Spearman positive correlation between both biomarkers (AChE and burrowing) suggested that inhibition of AChE activity was linked to behaviour impairments. At the population level, lower AChE and behaviour activities were detected in worms corresponding to lower population density and biomass. These results provide direct empirical field evidence demonstrating the sensitivity of behaviour of H. diversicolor as a biomonitor of estuarine health status assessment.

Metal stress-related gene expression patterns in two marine invertebrates, Hediste diversicolor (Annelida, Polychaeta) and Littorina littorea (Mollusca, Gastropoda), at a former mining site.

Abandoned mines often contaminate sediments with dissolved heavy metals and are known to impact many taxa. Physiological responses needed to avoid oxidative stress from metal toxicity include the upregulation of detoxification and metal-binding proteins such as glutathione-s-transferases and metallothioneins, which have been studied in diverse species. Fewer studies, however, have focused on gene expression changes to better understand these molecular mechanisms, especially across multiple species at a single contaminated site. To this end, the purpose of this study was to characterize metal stress-related gene expression in two species from different phyla, Hediste diversicolor (Annelida, Polychaeta) and Littorina littorea (Mollusca, Gastropoda), at a former mine site (Callahan Mine, Maine, USA). Both species and sediments were collected from a mine-affected tidal estuary (Goose Pond) and a nearby reference site. Elevated sediment metal levels were confirmed at Goose Pond. H. diversicolor individuals weighed significantly less at Goose Pond, while L. littorea weighed similarly at both sites. Transcript levels were stable in H. diversicolor but weakly upregulated in L. littorea, which likely reflect the importance of other physiological strategies for metal sequestration, or variable metal exposure at the individual level, respectively. In addition, patterns in glutathione-s-transferase expression differed across isoforms in H. diversicolor, while L. littorea exhibited divergent expression patterns in foot muscle and hepatopancreas. Overall, these results reinforce that diverse species likely undergo different physiological responses to metal toxicity, and more research is needed to investigate these mechanisms.

Effect of low frequency electromagnetic field on the behavior and bioenergetics of the polychaete Hediste diversicolor.

The aim of the present study was to determine the effect of an electromagnetic field (EMF) of value typically recorded in the vicinity of submarine cables (50 Hz, 1 mT) on the behavior and bioenergetics of the polychaete Hediste diversicolor. No avoidance or attraction behavior to EMF was shown, but the burrowing activity of H. diversicolor was enhanced in EMF treatment, indicating a stimulating effect on bioturbation potential. The polychaete maintained a positive energy balance and high amount (85% of assimilated energy) of energy available for individual production (scope for growth) after exposure to EMF. Food consumption and respiration rates were not affected but ammonia excretion rate was significantly reduced in EMF-exposed animals compared to the control conditions (geomagnetic field). The mechanisms behind this effect remain, however, unclear. This is the first study demonstrating the effects of environmentally realistic EMF value on the behavior and physiology of marine invertebrates, thus there is a need for more research.