Mechanism of barotaxis in marine zooplankton.

Hydrostatic pressure is a dominant environmental cue for vertically migrating marine organisms but the physiological mechanisms of responding to pressure changes remain unclear. Here, we uncovered the cellular and circuit bases of a barokinetic response in the planktonic larva of the marine annelid Platynereis dumerilii. Increased pressure induced a rapid, graded, and adapting upward swimming response due to the faster beating of cilia in the head multiciliary band. By calcium imaging, we found that brain ciliary photoreceptors showed a graded response to pressure changes. The photoreceptors in animals mutant for ciliary opsin-1 had a smaller sensory compartment and mutant larvae showed diminished pressure responses. The ciliary photoreceptors synaptically connect to the head multiciliary band via serotonergic motoneurons. Genetic inhibition of the serotonergic cells blocked pressure-dependent increases in ciliary beating. We conclude that ciliary photoreceptors function as pressure sensors and activate ciliary beating through serotonergic signalling during barokinesis.

Fauna - Microplastics interactions: Empirical insights from benthos community exposure to marine plastic waste.

Microplastic deposition in soft marine sediments raises concerns on their role in sediment habitats and unknown effects on resident macrobenthic communities. To assess the reciprocal influence that MPs and macrobenthos might have on each other, we performed a mesocosm experiment with ambient concentrations of environmental Polyethylene (PE) and a non-manipulated, natural macrobenthic community from the Belgian part of the North Sea (BPNS). Our results show that PE fragments increase mortality of abundant bivalves (specifically Abra alba) after 30 days of exposure but not for the most abundant polychaete Owenia fusiformis, possibly due to its predominant suspension feeding behavior. Fast burial of surface MPs exposes deep-dwelling burrowers to the pollutant, however reducing the amount of MPs interacting with (sub) surface living fauna. We conclude that macrobenthos promotes the sequestration of deposited MPs, counteracting resuspension, and can have cascading effects on biodiversity due to their effect on abundant and functionally important species.

Aggregative versus solitary settlement in Spirobranchus cariniferus gray, 1843 (serpulinae). What is the trade-off?

Sessile marine invertebrates usually follow a distinct pattern of living in dense aggregations or as solitary individuals. However, at least some serpulins, including Spirobranchus cariniferus, seem to be able to settle aggregative or solitary. To understand how living solitary or in aggregation is beneficial, it is essential to understand the advantages and disadvantages of both settlement strategies for sessile invertebrates. Benefits of living in aggregations include securing suitable habitat, improving the probability of survival by mitigating physical stress and increasing reproductive success. However, living in patches also comes with some disadvantages for the individual, such as higher intra- and interspecific competition for food, space and oxygen. Increased physiological stress can lead to increased mortality and decreased reproductive success, whereas solitary individuals could produce more gametes because of a lack of competition for food and space. On the other hand, predators would have easier access to them, and the possibility of fertilisation success may be lower because of a lack of synchronisation and a greater distance between individuals of different sexes. These issues have not been sufficiently addressed, particularly for serpulids. Individuals of the New Zealand endemic polychaete Spirobranchus cariniferus can be found solitary and aggregative in the same habitat. Therefore, this study is the first on serpulids comparing the growth and mortality of individuals living alone or in aggregations. Hence, bi-monthly observation of mortality and growth measurements were conducted on tagged individuals in the field, and weekly observations were conducted in a laboratory-based study on individuals of both settlement configurations. A final comparison of body metrics to tube dimensions was made by removing an individual from their tube. My findings revealed that while solitary and aggregative individuals elongate their tubes at a similar rate, further correlations of the body to tube sizes lead to the conclusion that solitary worms focus more of their energy on tube growth rather than body size increment than aggregative conspecifics. Mortality is highly variable and seems not to differ between both configurations. However, individuals living in a patch can better recover from damage to their tubes. Here presented observations hopefully initiated further studies into the effects of aggregation size and density on individual growth. Results of this and subsequent studies can inform the management efforts for reefs of serpulins, bivalves and other invertebrates.

Bacterial envelope polysaccharide cues settlement and metamorphosis in the biofouling tubeworm Hydroides elegans.

Metamorphosis for many marine invertebrates is triggered by external cues, commonly produced by bacteria. For larvae of Hydroides elegans, lipopolysaccharide (LPS) from the biofilm-dwelling bacterium Cellulophaga lytica induces metamorphosis. To determine whether bacterial LPS is a common metamorphosis-inducing factor for this species, we compare larval responses to LPS from 3 additional inductive Gram-negative marine biofilm bacteria with commercially available LPS from 3 bacteria not known to induce metamorphosis. LPS from all the inductive bacteria trigger metamorphosis, while LPS from non-inductive isolated marine bacteria do not. We then ask, which part of the LPS is the inductive element, the lipid (Lipid-A) or the polysaccharide (O-antigen), and find it is the latter for all four inductive bacteria. Finally, we examine the LPS subunits from two strains of the same bacterial species, one inductive and the other not, and find the LPS and O-antigen to be inductive from only the inductive bacterial strain.

A new genus and species of nudibranch-mimicking Syllidae (Annelida, Polychaeta).

Nudibranch mollusks, which are well-known for their vivid warning coloration and effective defenses, are mimicked by diverse invertebrates to deter predation through both Müllerian and Batesian strategies. Despite extensive documentation across different taxa, mimickers have not been detected among annelids, including polychaetes, until now. This study described a new genus and species of polychaete living on Dendronephthya octocorals in Vietnam and Japan. Belonging to Syllidae, it exhibits unique morphological adaptations such as a low number of body segments, simple chaetae concealed within the parapodia and large and fusiform antennae and cirri. Moreover, these appendages are vividly colored, featuring an internal dark red area with numerous terminal white spots and bright yellow tips, effectively contributing to mimicking the appearance of a nudibranch. This discovery not only documents the first known instance of such mimicry among annelids, but also expands our understanding of evolutionary adaptation and ecological strategies in marine invertebrates.

Variations in cell plasticity and proliferation underlie distinct modes of regeneration along the antero-posterior axis in the annelid Platynereis.

The capacity to regenerate lost tissues varies significantly among animals. Some phyla, such as the annelids, display substantial regenerating abilities, although little is known about the cellular mechanisms underlying the process. To precisely determine the origin, plasticity and fate of the cells participating in blastema formation and posterior end regeneration after amputation in the annelid Platynereis dumerilii, we developed specific tools to track different cell populations. Using these tools, we find that regeneration is partly promoted by a population of proliferative gut cells whose regenerative potential varies as a function of their position along the antero-posterior axis of the worm. Gut progenitors from anterior differentiated tissues are lineage restricted, whereas gut progenitors from the less differentiated and more proliferative posterior tissues are much more plastic. However, they are unable to regenerate the stem cells responsible for the growth of the worms. Those stem cells are of local origin, deriving from the cells present in the segment abutting the amputation plane, as are most of the blastema cells. Our results favour a hybrid and flexible cellular model for posterior regeneration in Platynereis relying on different degrees of cell plasticity.

Developmental stage dependent effects of posterior and germline regeneration on sexual maturation in Platynereis dumerilii.

Regeneration, regrowing lost and injured body parts, is an ability that generally declines with age or developmental transitions (i.e. metamorphosis, sexual maturation). Regeneration is also an energetically costly process, and trade-offs occur between regeneration and other costly processes such as growth, or sexual reproduction. Here we investigate the interplay of regeneration, reproduction, and developmental stage in the segmented worm Platynereis dumerilii. P. dumerilii can regenerate its whole posterior body axis, along with its reproductive cells, thereby having to carry out the two costly processes (somatic and germ cell regeneration) after injury. We specifically examine how developmental stage affects the success of germ cell regeneration and sexual maturation in developmentally young versus developmentally old organisms. We hypothesized that developmentally younger individuals (i.e. with gametes in early mitotic stages) will have higher regeneration success than the individuals at developmentally older stages (i.e. with gametes undergoing meiosis and maturation). Surprisingly, older amputated worms grew faster and matured earlier than younger amputees. To analyze germ cell regeneration during and after posterior regeneration, we used Hybridization Chain Reaction for the germline marker vasa. We found that regenerated worms start repopulating new segments with germ cell clusters as early as 14 days post amputation. In addition, vasa expression is observed in a wide region of newly-regenerated segments, which appears different from expression patterns during normal growth or regeneration in worms before gonial cluster expansion.

Combined effects of organic and mineral UV-filters on the lugworm Arenicola marina.

Pollution from personal care products, such as UV-filters like avobenzone and nano-zinc oxide (nZnO), poses a growing threat to marine ecosystems. To better understand this hazard, especially for lesser-studied sediment-dwelling marine organisms, we investigated the physiological impacts of simultaneous exposure to nZnO and avobenzone on the lugworm Arenicola marina. Lugworms were exposed to nZnO, avobenzone, or their combination for three weeks. We assessed pollutant-induced metabolic changes by measuring key metabolic intermediates in the body wall and coelomic fluid, and oxidative stress by analyzing antioxidant levels and oxidative lesions in proteins and lipids of the body wall. Exposure to UV filters resulted in shifts in the concentrations of Krebs' cycle and urea cycle intermediates, as well as alterations in certain amino acids in the body wall and coelomic fluid of the lugworms. Pathway enrichment analyses revealed that nZnO induced more pronounced metabolic shifts compared to avobenzone or their combination. Exposure to avobenzone or nZnO alone prompted an increase in tissue antioxidant capacity, indicating a compensatory response to restore redox balance, which effectively prevented oxidative damage to proteins or lipids. However, co-exposure to nZnO and avobenzone suppressed superoxide dismutase and lead to accumulation of lipid peroxides and methionine sulfoxide, indicating oxidative stress and damage to lipids and proteins. Our findings highlight oxidative stress as a significant mechanism of toxicity for both nZnO and avobenzone, especially when combined, and underscores the importance of further investigating the fitness implications of oxidative stress induced by these common UV filters in benthic marine organisms.

Metal bioaccumulation and effects of olivine sand exposure on benthic marine invertebrates.

Due to the anthropogenic increase of atmospheric CO2 emissions, humanity is facing the negative effects of rapid global climate change. Both active emission reduction and carbon dioxide removal (CDR) technologies are needed to meet the Paris Agreement and limit global warming to 1.5 °C by 2050. One promising CDR approach is coastal enhanced weathering (CEW), which involves the placement of sand composed of (ultra)mafic minerals like olivine in coastal zones. Although the large-scale placement of olivine sand could beneficially impact the planet through the consumption of atmospheric CO2 and reduction in ocean acidification, it may also have physical and geochemical impacts on benthic communities. The dissolution of olivine can release dissolved constituents such as trace metals that may affect marine organisms. Here we tested acute and chronic responses of marine invertebrates to olivine sand exposure, as well as examined metal accumulation in invertebrate tissue resulting from olivine dissolution. Two different ecotoxicological experiments were performed on a range of benthic marine invertebrates (amphipod, polychaete, bivalve). The first experiment included acute and chronic survival and growth tests (10 and 20 days, respectively) of olivine exposure while the second had longer (28 day) exposures to measure chronic survival and bioaccumulation of trace metals (e.g. Ni, Cr, Co) released during olivine sand dissolution. Across all fauna we observed no negative effects on acute survival or chronic growth resulting solely from olivine exposure. However, over 28 days of exposure, the bent-nosed clam Macoma nasuta experienced reduced burrowing and accumulated 4.2 ± 0.7 µg g ww-1 of Ni while the polychaete Alitta virens accumulated 3.5 ± 0.9 µg g ww-1 of Ni. No significant accumulation of any other metals was observed. Future work should include longer-term laboratory studies as well as CEW field studies to validate these findings under real-world scenarios.

Secondary-tail formation during stolonization in the Japanese green syllid, Megasyllis nipponica.

Benthic annelids belonging to the family Syllidae show a distinctive sexual reproduction mode called "stolonization," in which posterior segments are transformed into a reproductive individual-like unit called a "stolon." Megasyllis nipponica forms a stolon head and a secondary tail in the middle of the trunk before a stolon detaches, while, in the case of posterior amputation, posterior regeneration initiates at the wound after amputation. To understand the difference between posterior regeneration and secondary-tail formation during stolonization, detailed comparisons between the developmental processes of these two tail-formation types were performed in this study. Morphological and inner structural observations (i.e., cell proliferation and muscular/nervous development) showed that some processes of posterior regeneration, such as blastema formation and muscular/nervous regeneration at the amputation site, are missing during secondary-tail formation. In contrast, the secondary tail showed some unique features, such as the formation of ventrolateral half-tail buds that later fused in the middle and muscle/nerve branches formed before the detachment of the stolon. These novel features in the process of stolonization are suggested to be adaptive since the animals need to recover a posterior end quickly to stolonize again.

Influence of habitat features on the colonisation of native and non-indigenous species.

Marine artificial structures provide substrates on which organisms can settle and grow. These structures facilitate establishment and spread of non-indigenous species, in part due to their distinct physical features (substrate material, movement, orientation) compared to natural habitat analogues such as rocky shores, and because following construction, they have abundant resources (space) for species to colonise. Despite the perceived importance of these habitat features, few studies have directly compared distributions of native and non-indigenous species or considered how functional identity and associated environmental preferences drive associations. We undertook a meta-analysis to investigate whether colonisation of native and non-indigenous species varies between artificial structures with features most closely resembling natural habitats (natural substrates, fixed structures, surfaces oriented upwards) and those least resembling natural habitats (artificial materials, floating structures, downfacing or vertical surfaces), or whether functional identity is the primary driver of differences. Analyses were done at global and more local (SE Australia) scales to investigate if patterns held regardless of scale. Our results suggest that functional group (i.e., algae, ascidians. barnacles, bryozoans, polychaetes) rather than species classification (i.e., native or non-indigenous) are the main drivers of differences in communities between different types of artificial structures. Specifically, there were differences in the abundance of ascidians, barnacles, and polychaetes between (1) upfacing and downfacing/vertical surfaces, and (2) floating and fixed substrates. When differences were detected, taxa were most abundant on features least resembling natural habitats. Results varied between global and SE Australian analyses, potentially due to reduced variability across studies in the SE Australian dataset. Thus, the functional group and associated preferences of the highest threat NIS in the area should be considered in design strategies (e.g., ecological engineering) to limit their establishment on newly built infrastructure.

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.

Temperature and sex shape reproductive barriers in a climate change hotspot.

Climate change is altering species ranges and reproductive interactions in existing ranges, offering species new scope to mate and hybridize. The outcomes will depend on how environmental factors shape reproductive barriers across life stages, yet this is rarely assessed across the environments that species encounter in nature. We assess prezygotic and postzygotic barriers, and their dependence on temperature and parental sex, in species of a reef-building tubeworm (Galeolaria) from a fast-warming biodiversity hotspot in southern Australia. By replicating pure and reciprocal hybrid crosses across 5 temperatures spanning species' thermal ranges, we estimate thermal tolerance curves (defining niches) for crosses and reproductive isolation at each temperature. By also replicating crosses at 3 life stages, we partition the contributions of prezygotic barriers at fertilization, postzygotic barriers at embryogenesis, and postzygotic barriers at larval development to reproductive isolation. We show that barriers are weaker at fertilization and embryogenesis, but stronger and more temperature sensitive at larval development, as species diverge in thermal niche. Asymmetry of barriers between parental sexes, moreover, suggests a complex interplay between niche differentiation and maternal inheritance. Our findings point to a key role for temperature in reproductive isolation, but also challenges for predicting the fate of isolation in future climates.

Identification and expression analysis of Oxfibrillin gene involved in the regeneration process of Ophryotrocha xiamen (Annelida, Dorcilleidae).

Regeneration of lost body parts is a widespread phenomenon across annelids. However, the molecular inducers of the cell sources for this reparative morphogenesis have not been identified. We have identified a regeneration-related gene Oxfibrillin from the transcriptome analysis of a polychaeta, Ophryotrocha xiamen, which is found to be a well-suited model to study the mechanisms of regeneration. Fibrillins are large glycoproteins that assemble to form the microfibrils and regulate growth factors or other transfer processes. Here, we obtained the 31,274 bp genomic DNA sequences of Oxfibrillin. The coding sequence length was 5784 bp encoding 1927 amino acids with a VWD domain, EGF/cb-EGF domains, a TR domain, and a transmembrane domain. Oxfibrillin was positioned within the subgroup of invertebrates and showed low scores for homology to mammalian fibrillin. In gene expression analysis, Oxfibrillin genes were constantly upregulated during the early regeneration process and then remained stable until the formation of the complete tail which indicated that it might be a vital factor to affect posterior regeneration process. Therefore, the Oxfibrillin of O. xiamen might play important roles in the regeneration process.

Avoiding UV light.

The larvae of an annelid worm use nitric oxide signalling to activate the neural pathways needed to swim away from the harmful ultraviolet light of the sun.

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.

Cell Proliferation Indices in Regenerating Alitta virens (Annelida, Errantia).

In recent years, interest in the possible molecular regulators of cell proliferation and differentiation in a wide range of regeneration models has grown significantly, but the cell kinetics of this process remain largely a mystery. Here we try to elucidate the cellular aspects of regeneration by EdU incorporation in intact and posteriorly amputated annelid Alitta virens using quantitative analysis. We found that the main mechanism of blastema formation in A. virens is local dedifferentiation; mitotically active cells of intact segments do not significantly contribute to the blastemal cellular sources. Amputation-induced proliferation occurred predominantly within the epidermal and intestinal epithelium, as well as wound-adjacent muscle fibers, where clusters of cells at the same stage of the cell cycle were found. The resulting regenerative bud had zones of high proliferative activity and consisted of a heterogeneous population of cells that differed in their anterior-posterior positions and in their cell cycle parameters. The data presented allowed for the quantification of cell proliferation in the context of annelid regeneration for the first time. Regenerative cells showed an unprecedentedly high cycle rate and an exceptionally large growth fraction, making this regeneration model especially valuable for studying coordinated cell cycle entry in vivo in response to injury.

Future research directions of the model marine tubeworm Hydroides elegans and synthesis of developmental staging of the complete life cycle.

BACKGROUND: The biofouling marine tube worm, Hydroides elegans, is an indirect developing polychaete with significance as a model organism for questions in developmental biology and the evolution of host-microbe interactions. However, a complete description of the life cycle from fertilization through sexual maturity remains scattered in the literature, and lacks standardization. RESULTS AND DISCUSSION: Here, we present a unified staging scheme synthesizing the major morphological changes that occur during the entire life cycle of the animal. These data represent a complete record of the life cycle, and serve as a foundation for connecting molecular changes with morphology. CONCLUSIONS: The present synthesis and associated staging scheme are especially timely as this system gains traction within research communities. Characterizing the Hydroides life cycle is essential for investigating the molecular mechanisms that drive major developmental transitions, like metamorphosis, in response to bacteria.

Rapid manoeuvre of fan worms (Annelida: Sabellidae) through tubes.

Multiple variables determine the success of an escape response of an animal, and the rapidity of the escape manoeuvre is often the most important. Fan worms (Annelida: Sabellidae) can rapidly withdraw their tentacles, which are covered in heavily ciliated ramifications called pinnules, into their tubes to protect them from approaching threats. Here, we explore the dynamic and mechanistic features behind this escape manoeuvre. The escape responses of fan worms were recorded by high-speed videography and quantified by computerized motion analysis, showing an ultrahigh retraction speed of 272±135 mm s-1 (8±4 body lengths s-1). We found that fan worms possess powerful muscle-driven systems, which can generate contractive forces up to 36 times their body weight. In order to achieve these rapid, forceful movements through seawater without damaging their tentacles, fan worms have developed functional morphological adaptations to reduce fluidic drag, including the flattening of their radiolar pinnules and the deformation of bodily segmental ridges. Our hydrodynamic models indicate that these mechanical processes can decrease fluidic drag by 47%, trapped mass by 75% and friction coefficient by 89%. These strategies allow fan worms to execute rapid escape responses and could inspire the design of fast in-pipe robots.

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.