Sulfur, sterol and trehalose metabolism in the deep-sea hydrocarbon seep tubeworm Lamellibrachia luymesi.

BACKGROUND: Lamellibrachia luymesi dominates cold sulfide-hydrocarbon seeps and is known for its ability to consume bacteria for energy. The symbiotic relationship between tubeworms and bacteria with particular adaptations to chemosynthetic environments has received attention. However, metabolic studies have primarily focused on the mechanisms and pathways of the bacterial symbionts, while studies on the animal hosts are limited. RESULTS: Here, we sequenced the transcriptome of L. luymesi and generated a transcriptomic database containing 79,464 transcript sequences. Based on GO and KEGG annotations, we identified transcripts related to sulfur metabolism, sterol biosynthesis, trehalose synthesis, and hydrolysis. Our in-depth analysis identified sulfation pathways in L. luymesi, and sulfate activation might be an important detoxification pathway for promoting sulfur cycling, reducing byproducts of sulfide metabolism, and converting sulfur compounds to sulfur-containing organics, which are essential for symbiotic survival. Moreover, sulfide can serve directly as a sulfur source for cysteine synthesis in L. luymesi. The existence of two pathways for cysteine synthesis might ensure its participation in the formation of proteins, heavy metal detoxification, and the sulfide-binding function of haemoglobin. Furthermore, our data suggested that cold-seep tubeworm is capable of de novo sterol biosynthesis, as well as incorporation and transformation of cycloartenol and lanosterol into unconventional sterols, and the critical enzyme involved in this process might have properties similar to those in the enzymes from plants or fungi. Finally, trehalose synthesis in L. luymesi occurs via the trehalose-6-phosphate synthase (TPS) and trehalose-6-phosphate phosphatase (TPP) pathways. The TPP gene has not been identified, whereas the TPS gene encodes a protein harbouring conserved TPS/OtsA and TPP/OtsB domains. The presence of multiple trehalases that catalyse trehalose hydrolysis could indicate the different roles of trehalase in cold-seep tubeworms. CONCLUSIONS: We elucidated several molecular pathways of sulfate activation, cysteine and cholesterol synthesis, and trehalose metabolism. Contrary to the previous analysis, two pathways for cysteine synthesis and the cycloartenol-C-24-methyltransferase gene were identified in animals for the first time. The present study provides new insights into particular adaptations to chemosynthetic environments in L. luymesi and can serve as the basis for future molecular studies on host-symbiont interactions and biological evolution.

Quantitative comparison of clamworm (Perinereis aibuhitensis) and crab (Macrophthalmus japonicus) burrowing effects on nitrogen and phosphorus dynamics at the sediment-water interface.

Exploring nitrogen and phosphorus dynamics in estuarine ecosystems is of great significance for understanding the nutrient cycle in terrestrial and marine ecosystems. However, the ecological functions of common species remain unclear because there is a dearth of observational data. To quantitatively compare the bioturbation effects of the clamworm Perinereis aibuhitensis and crab Macrophthalmus japonicus on the migration of nitrogen and phosphorus in the sediment-pore water-overlying water interfaces, an indoor experiment was carried out using microcosms containing substrate, water, and living specimens collected from the Dagu Estuary, China. The physical structure of the sediments was greatly altered by both clamworms and crabs, and the amount of suspended particulate matter in the overlying water was in the order: crab > clamworm > control groups. Sediments, especially the 0-2 cm layer, were continuously mineralized under bioturbation, releasing NH3-N, NO2--N, NO3--N, and dissolved total phosphorus. During the experiment, the release rate of NH3-N (2.45 mg L-1 to 10.23 mg L-1), NO2--N (0.25 mg L-1 to 7.34 mg L-1), NO3--N (0.02 mg L-1 to 6.03 mg L-1), and dissolved total phosphorus (0.07 mg L-1 to 0.16 mg L-1) in the crab group was higher than that in the clamworm group (NH3-N, 0.93 mg L-1 to 6.11 mg L-1; NO2--N, 0.51 mg L-1 to 6.18 mg L-1; NO3--N, 0.01 mg L-1 to 5.24 mg L-1, and dissolved total P, 0.04 mg L-1 to 0.08 mg L-1). Our findings indicate that crabs might play a more important role in promoting nitrogen and phosphorus migration than clamworms because of their stronger bioturbation strategies in sediments. Bioturbation by both clamworms and crabs reduced nitrogen and phosphorus accumulation in surface sediments and accelerated organic matter mineralization in the sediments, which may promote nutrient recycling in estuarine ecosystems.

Phytoremediation of crude oil-contaminated sediment using Suaeda heteroptera enhanced by Nereis succinea and oil-degrading bacteria.

A 150-day experiment was performed to investigate the stimulatory effect of a promising phytoremediation strategy consisting of Suaeda heteroptera (S. heteroptera), Nereis succinea (N. succinea), and oil-degrading bacteria for cleaning up total petroleum hydrocarbons (TPHs) in spiked sediment. Inoculation with oil-degrading bacteria and/or N. succinea increased plant yield and TPH accumulation in S. heteroptera plants. The highest TPH dissipation (40.5%) was obtained in the combination treatment, i.e., S. heteroptera + oil-degrading bacteria + N. succinea, in which the sediment TPH concentration decreased from an initial value of 3955 to 2355 mg/kg in 150 days. BAF, BCF, and TF confirmed the role of N. succinea and oil-degrading bacteria in the amelioration and translocation of TPHs. In addition, TPH toxicity of S. heteroptera was alleviated by N. succinea and oil-degrading bacteria addition through the reduction of oxidative stress. Therefore, S. heteroptera could be used for cleaning up oil-contaminated sediment, particularly in the presence of oil-degrading bacteria + N. succinea. Field studies on oil-degrading bacteria + N. succinea may provide new insights on the rehabilitation and restoration of sediments contaminated by TPHs.

Our study attempted to investigate the stimulatory effect of a promising phytoremediation strategy consisting of Suaeda heteroptera (S. heteroptera), Nereis succinea (N. succinea), and oil-degrading bacteria for cleaning up TPH in spiked sediment. Planting S. heteroptera can greatly increase sediment TPH removal, and its removal was enhanced greater after inoculation with oil-degrading bacteria and/or N. succinea. Moreover, the promising phytoremediation strategy developed in the current work can serve as an efficient, novel approach to removal TPH in sediment/soil. In our opinions, these findings provide insights into the assessment of their ecological risks in the environments that are of interest to broad readership of International Journal of Phytoremediation.

Endogenous Fluorescent Proteins in the Mucus of an Intertidal Polychaeta: Clues for Biotechnology.

The vast ocean holds many unexplored organisms with unique adaptive features that enable them to thrive in their environment. The secretion of fluorescent proteins is one of them, with reports on the presence of such compounds in marine annelids being scarce. The intertidal Eulalia sp. is an example. The worm secretes copious amounts of mucus, that when purified and concentrated extracts, yield strong fluorescence under UV light. Emission has two main maxima, at 400 nm and at 500 nm, with the latter responsible for the blue-greenish fluorescence. Combining proteomics and transcriptomics techniques, we identified ubiquitin, peroxiredoxin, and 14-3-3 protein as key elements in the mucus. Fluorescence was found to be mainly modulated by redox status and pH, being consistently upheld in extracts prepared in Tris-HCl buffer with reducing agent at pH 7 and excited at 330 nm. One of the proteins associated with the fluorescent signal was localized in secretory cells in the pharynx. The results indicate that the secretion of fluorescent proteinaceous complexes can be an important defense against UV for this dweller. Additionally, the internalization of fluorescent complexes by ovarian cancer cells and modulation of fluorescence of redox status bears important considerations for biotechnological application of mucus components as markers.

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.

Responses of the macrobenthic community to the Dalian Bay oil spill based on co-occurrence patterns and interaction networks.

On July 16, 2010, a pipeline explosion spilled 1500 tons of crude oil into the Port of Dalian, China. To identify taxa responses to the spill, we exploited seven years of monitoring data to examine the co-occurrence of taxon pairs and the variation of the macrobenthic community. Non-parametric correlation analysis was used to construct interaction networks of relationships between oil spill contaminants and macrobenthic taxa. We observed that the impacted macrobenthic community not restored before 2016. The tolerance/sensitivity of taxa was inconsistent with the studies of oil impacts in other locations. We suggest revision of the ecological group classification of Sabellidae, Lumbrineridae, Terebellidae, Sternaspidae, and Spionidae. The variation in the frequency of coexistence indicates the potential impact of oil spill pollution on resource occupation. The interaction network involving macrobenthic families and stressors associated with the oil spill highlights how different macrobenthic families respond to different combinations of stressors.

Anti-fatigue effect of aqueous extract of Hechong (Tylorrhynchus heterochaetus) via AMPK linked pathway.

Tylorrhynchus heterochaetus (Hechong in Chinese) has been used in Chinese traditional medicine for treating various diseases. This study was aimed to assess the anti-fatigue effect of T. heterochaetus on Kunming mice and its primary mechanism of action using forced running, rotating rod and weight-loaded swimming tests. Low (2.70 mg/0.5 mL/20 g), medium (5.41 mg/0.5 mL/20 g) and high (6.58 mg/0.5 mL/20 g) doses of T. heterochaetus aqueous extract were treated to mice for 28 days. Among the doses, the low and medium doses showed significant (p ≤ 0.05) anti-fatigue effect on the weight-loaded swimming test. Also, T. heterochaetus extract showed significant (p ≤ 0.05) effects on fatigue-related blood parameters by increasing the GLU, TG and LDH levels and decreasing the LA, CK and BUN levels. The levels of liver and skeletal muscle glycogen were also significantly (p ≤ 0.05) increased after treatment. Further, on Western blot analysis, it has been found that T. heterochaetus enhanced the expressions of AMPK and PGC-1α in the liver and skeletal muscles of mice. From the study, our outcomes suggest that T. heterochaetus possess an anti-fatigue effect through the AMPK-linked pathway and thereby it can regularize the energy metabolism.

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.

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.

Analysis of petroleum biodegradation by a bacterial consortium isolated from worms of the polychaeta class (Annelida): Implications for NPK fertilizer supplementation.

Degradation of petroleum hydrocarbons using bacterial consortia may be a means of optimizing bioremediation techniques. In this study, bacterial strains were isolated from the digestive tract of polychaete worms and evaluated concerning the potential of the bacteria to degrade petroleum compounds (Acinetobacter sp., Bacillus sp., Pantoea sp. and Enterobacter sp.). The strains were separately screened regarding their potential to degrade oil after 24 h. The main experiment was carried out for 30 days with the addition of nitrogen, phosphorus and potassium (NPK) after 15 days (Bact-NPK15) and 28 days (Bact-NPK28) and without NPK (Bact). The Bact treatment biomass remained stable until the 20th day of the experiment. C13-C40 n-alkanes were degraded with all treatments in the following order: Bact>Bact-NPK28>Bact-NPK15. Significant differences were observed between the controls and all treatments (p = 0.00031). Measurement of polycyclic aromatic hydrocarbons (PAHs) indicates a lower contribution of these compounds in the Bact-NPK28 treatment, although no significant difference between groups was observed. Bact-NPK28 was able to remove 40% of naphthalene, while Bact-NPK15 removed 20%; this effect was not observed in Bact. Higher hopane degradation levels were observed in Bact and, to a lesser extent, in Bact-NPK28. NPK application for 28 days mainly favored PAH degradation. The evaluated consortium thus exhibits potential in the bioremediation of petroleum-contaminated areas.

Effects of mixtures of anticancer drugs in the benthic polychaete Nereis diversicolor.

The increasing consumption of anticancer drugs through single and/or combinatory chemotherapy worldwide raised concern regarding their toxicity burden in coastal zones. The toxicity of a mixture of three compounds involving the drugs cisplatin (CisPt), cyclophosphamide (CP) and tamoxifen (TAM) was determined on the marine polychaete Nereis diversicolor exposed to an increasing range of their concentrations, respectively: Mix A: 0.1 + 10 + 0.1 ng L-1; Mix B: 10 + 100 + 10 ng L-1; Mix C: 100 + 500 + 25 ng L-1; Mix D: 100 + 1000 + 100 ng L-1. Different endpoints were assessed, including disturbance in the burrowing behaviour, neurotoxicity (acetylcholinesterase - AChE activity), antioxidant enzymes (superoxide dismutase - SOD; catalase - CAT; selenium-dependent glutathione peroxidase - Se-GPx and total glutathione peroxidases T-GPx activities), biotransformation metabolism (glutathione-S-transferases - GST), lipid peroxidation (LPO) and genotoxicity (DNA damage). Biological effects of the mixtures of anticancer compounds on N. diversicolor were compared with previous studies about effects on the same biological model under single-drug exposure conducted with the same molecules. Regarding SOD activity, TAM showed an antagonist effect over CisPt and CP in mixtures C and D. In Mix D, there was a synergistic effect of TAM and CisPt that inhibited CAT activity and an additive interaction of CisPt and CP on the Phase II biotransformation enzyme. Drugs in Mix A also suppressed polychaetes' GST activity, although different from the respective single-drug responses, besides able to induce T-GPx activity, that was not sufficient to avoid oxidative damage and mid-grade DNA damage. Due to the absence of burrowing impairment in Mix A, mechanisms involved in neurotoxicity were other than the one driven by AChE alterations. At the intermediary concentrations (Mix B and C), only LPO occurred. Data from drugs individually may not predict the risks provided by mixtures.

Enhanced removal of sediment-associated total petroleum hydrocarbons under bioturbation by polychaete perinereis aibuhitensis.

Bioturbation processes could influence the physical, chemical and biological properties of aquatic sediments and improve the mineralization of organic matter in sediment. The influence of bioturbation by polychaete Perinereis aibuhitensis on the removal of sediment-associated total petroleum hydrocarbon (TPH) was evaluated through laboratory experiment with P. aibuhitensis cultured in crude oil contaminated coastal sediment. After 60 days, the TPH concentrations in the sediments were significantly decreased compared to the initial concentrations, in which the TPH concentrations in bioturbation experiments (with worms) were significantly lower than those in control experiments (without worms) for both low (1.48 ± 0.19g/kg dry wt) and high (2.67 ± 0.33 g/kg dry wt) TPH-contaminated groups, indicating bioturbation enhanced the removal of TPH in sediment. The TPH removal rates in high TPH group were significantly lower than those in low TPH group, suggested that petroleum pollution inhibited the degradation of petroleum hydrocarbons in sediment. However, the stimulation efficiency was higher in high TPH group than that in low TPH group, which may be the result of enhanced hydrocarbon's bioavailability by digestive fluid during gut transit.

Cytotoxicity and enzymatic biomarkers as early indicators of benthic responses to the soluble-fraction of diesel oil.

Xenobiotics from oil tanker leaks and industrial discharges are amongst the main human impacts to confined coastal areas. We assessed the genotoxic responses to the water-soluble fraction of diesel oil in the polychaete Laeonereis culveri and the bivalve Anomalocardia flexuosa, two widespread benthic species in subtropical estuaries from the Southwestern Atlantic. We hypothesized that the highest responsiveness would be expressed by significantly different biomarkers responses between control and oil-impacted treatments. Responsiveness to diesel oil was investigated using an experimental design with two fixed factors (contaminant percentages and times of exposure). After exposure, we monitored the responses of the oxidative stress enzymes and performed micronuclei tests. Results were congruent for both species. Antioxidant defense of glutathione S-transferase and the induction of micronuclei and nuclear buds, the latter just for the bivalve, were significantly affected by polycyclic aromatic hydrocarbons, with significant increases on the seventh day of exposure and in the higher concentrations, compared to controls groups. We assessed the benefits and drawbacks of using each biomarker in laboratory experiments. Both species are indicators of early, and rapid responses to genotoxic contaminants in subtropical estuarine habitats. We suggest that the micronuclei frequency in A. flexuosa is a simple, fast and cheap test for genotoxicity in oil-impacted areas. Such early biomarkers are needed to develop better protocols for impact assessment and monitoring under real field conditions.

Effects of an invasive polychaete on benthic phosphorus cycling at sea basin scale: An ecosystem disservice.

Macrofaunal activities in sediments modify nutrient fluxes in different ways including the expression of species-specific functional traits and density-dependent population processes. The invasive polychaete genus Marenzelleria was first observed in the Baltic Sea in the 1980s. It has caused changes in benthic processes and affected the functioning of ecosystem services such as nutrient regulation. The large-scale effects of these changes are not known. We estimated the current Marenzelleria spp. wet weight biomass in the Baltic Sea to be 60-87 kton (95% confidence interval). We assessed the potential impact of Marenzelleria spp. on phosphorus cycling using a spatially explicit model, comparing estimates of expected sediment to water phosphorus fluxes from a biophysical model to ecologically relevant experimental measurements of benthic phosphorus flux. The estimated yearly net increases (95% CI) in phosphorous flux due to Marenzelleria spp. were 4.2-6.1 kton based on the biophysical model and 6.3-9.1 kton based on experimental data. The current biomass densities of Marenzelleria spp. in the Baltic Sea enhance the phosphorus fluxes from sediment to water on a sea basin scale. Although high densities of Marenzelleria spp. can increase phosphorus retention locally, such biomass densities are uncommon. Thus, the major effect of Marenzelleria seems to be a large-scale net decrease in the self-cleaning capacity of the Baltic Sea that counteracts human efforts to mitigate eutrophication in the region.

The biogenic reefs formed by the alien polychaete Hydroides dianthus (Serpulidae, Annelida) favor the polyp stage of Aurelia coerulea (Cnidaria, Scyphozoa) in a coastal artificial lake.

Blooms of the moon jellyfish Aurelia coerulea frequently occur in coastal waters. The increased availability of substrates for the settlement and proliferation of polyps due to the expansion of artificial structures in coastal areas has been proposed as a possible contributing factor in jellyfish blooms. This paper investigates whether a marine artificial lake (Fenghuang Lake) provides additional substrates for A. coerulea polyps and contributes to jellyfish blooms. High densities of A. coerulea ephyrae were discovered in this lake, with a mean density of 41 individuals/m3 and a maximum measured density of 128 individuals/m3. Meanwhile, A. coerulea ephyrae were also found in the two emptying channels outside the lake, with a mean density of 13 individuals/m3. Underwater surveys revealed that dense colonies of A. coerulea polyps occurred mainly on biogenic reefs formed by a polychaete, which was identified as an invasive serpulid species Hydroides dianthus, based on the phylogenetic analysis of mitochondrial COI gene sequences. Our study highlights the potential modification of habitats by the alien polychaete H. dianthus, which might provide complex benthic habits suitable for the settlement and proliferation of A. coerulea polyps and may contribute to jellyfish blooms in the marine artificial lake and nearby coastal waters.

Biomarker responses to environmental contamination in estuaries: A comparative multi-taxa approach.

Estuaries are highly productive ecosystems subjected to numerous anthropogenic pressures with consequent environmental quality degradation. In this study, multiple biomarker responses [superoxide dismutase (SOD), catalase (CAT), ethoxyresorufin O-deethylase (EROD) and glutathione S-transferase (GST) activities, as well as lipid peroxidation (LPO) and DNA damage (DNAd)] were determined in two fish (Dicentrarchus labrax and Pomatoschistus microps) and four macroinvertebrate species (Carcinus maenas, Crangon crangon, Hediste diversicolor and Scrobicularia plana) from the Ria de Aveiro and Tejo estuaries over distinct months. Two sites per estuarine system were selected based on anthropogenic pressures and magnitude of environmental contamination. Antioxidant enzyme activities in fish species suggested a ubiquitous response to oxidative stress, while biotransformation and effect biomarkers exhibited higher spatial and temporal variation. In invertebrate species, biotransformation enzyme activity was clearly less variable than in fish evidencing lower xenobiotic transformation capability. Overall, largest biomarker responses were found in the most contaminated sites (Tejo), yet species-specific patterns were evident. These should be factored in multi-taxa approaches, considering that the differential functional traits of species, such as habitat use, life-stage, feeding or physiology can influence exposure routes and biomarker responses. The Integrated Biomarker Response index highlighted patterns in biomarker responses which were not immediately evident when analyzing biomarkers individually. Overall, results provided insights into the complexity of species responses to contamination in naturally varying estuarine environments. Ultimately, multi-taxa and multi-biomarker approaches provide a comprehensive and complementary view of ecosystem health, encompassing diverse forms of biological integration and exposure routes, and allow the validation of results among markers and species.

New species for the biomitigation of a super-intensive marine fish farm effluent: Combined use of polychaete-assisted sand filters and halophyte aquaponics.

The main objective of this study was to test an innovative biomitigation approach, where polychaete-assisted (Hediste diversicolor) sand filters were combined with the production of Halimione portulacoides in aquaponics, to remediate an organic-rich effluent generated by a super intensive fish farm operating a land-based RAS (Recirculating aquaculture system). The set up included four different experimental combinations that were periodically monitored for 5months. After this period, polychaete-assisted sand filters reduced in 70% the percentage of OM and the average densities increased from ≈400ind.m-2 to 7000ind.m-2. H. portulacoides in aquaponics contributed to an average DIN (Dissolved inorganic Nitrogen) decrease of 65%, which increased to 67% when preceded by filter tanks stocked with polychaetes. From May until October (5months) halophytes biomass increased from 1.4kgm-2±0.7 (initial wet weight) to 18.6kgm-2±4.0. Bearing in mind that the uptake of carbon is mostly via photosynthesis and not though the uptake of dissolved inorganic carbon, this represents an approximate incorporation of ≈1.3kgm-2 carbon (C), ≈15gm-2 nitrogen (N) and ≈8gm-2 phosphorus (P) in the aerial part (76% of total biomass), and an approximate incorporation of ≈0.5kgm-2 carbon (C), ≈3gm-2 nitrogen (N) and ≈2gm-2 phosphorus (P) in the roots (24% of total biomass). In the present study, the potential of the two extractive species for biomitigation of a super-intensive marine fish farm effluent could be clearly demonstrated, contributing in this way to potentiate the implementation of more sustainable practices.

Molecular cloning and characterization of an estrogen receptor gene in the marine polychaete Perinereis aibuhitensis.

Estrogen receptors (ERs) are the primary mediators of estrogen signaling, and play crucial roles in the reproduction and development of vertebrates. The full-length cDNA of Perinereis aibuhitensis estrogen receptor (paER) was cloned and characterized for the first time. The positions of the cysteine residues and the residues around them, which constitute two zinc finger motifs and a P-box, are conserved in both vertebrates and invertebrates. A phylogenetic analysis revealed that paER is an orthologue of ER in the polychaete Platynereis dumerilii. A tissue distribution analysis of paER mRNA showed that it is expressed in various tissues, including the body wall, head, esophageal gland, esophagus, stomach, and most strongly in the intestines. Its expression was also measured in P. aibuhitensis after exposure to 17ß-estradiol (E2) for 48h. The paER mRNA levels in the body wall were measured after 6, 12, 24, and 48h in E2-exposed and control animals. However, no significant differences in paER expression were observed between them at any time point. This report describes the first molecular characterization of full-length paER and its tissue-specific expression in P. aibuhitensis.

Cu/Zn superoxide dismutase (SOD) and catalase (CAT) response to crude oil exposure in the polychaete Perinereis aibuhitensis.

Cu/Zn superoxide dismutase (SOD) and catalase (CAT) cDNAs from the polychaete Perinereis aibuhitensis were cloned and characterized in order to investigate the relationship between crude oil exposure and stress response in this worm. The full length of PaSOD was 870 bp and PaCAT was 1967 bp encoding 150 and 506 amino acids, respectively. Gene expression and enzyme activity of Cu/Zn SOD and CAT in response to crude oil contaminated soil (500, 1500, and 3000 mg/kg) were measured. The results showed that expression of the CAT gene and enzyme activity in P. aibuhitensis was positively correlated to the concentration of crude oil and reached a maximum at 15 days of exposure to 3000 mg/kg crude oil. The expression of the SOD gene and enzyme activity of SOD in P. aibuhitensis also increased during exposure to crude oil and reached a maximum at 10 days of exposure to 3000 mg/kg crude oil. These results indicated that SOD and CAT are important for maintaining the balance of cellular metabolism and protecting P. aibuhitensis from crude oil toxicity.

The integrated biomarker response: a suitable tool to evaluate toxicity of metal-based nanoparticles.

Nanotechnology is a much promising field of science and technology with applications in a wide range of areas such as electronics, biomedical applications, energy and cosmetics. Metal-based engineered nanoparticles (ENPs) are common in many technological applications; some of the most common nanoparticles available commercially are silver, gold, copper oxide (CuO), zinc oxide (ZnO) and cadmium sulphide (CdS). The toxicity of metal-based NPs may be either due to their specific physical characteristics as NPs or to the specific toxicity of metals released from NPs under environmental conditions. In this study we evaluated the toxicity effects of a range of ENPs (Ag, Au, CuO, CdS, ZnO) along with a control containing equivalent quantities of dissolved metal on two endobenthic species: the ragworm Hediste diversicolor and the bivalve Scrobicularia plana. A suite of complementary biomarkers was used to reveal toxicity effects. A common challenge in multibiomarkers studies is to go beyond an independent interpretation of each one, and to assess a global response of individuals. The Integrated Biomarker Response (IBR) was calculated for both species exposed to the different metal-based ENPs studied or to their dissolved metal counterpart to provide efficient and easy tools for environmental managers. We evidence that metal-based NPs lead to an overall difference in biological responses from that of their dissolved counterparts. The IBR could thus be considered as an efficient tool to transfer research results to stakeholders with possible implementation for regulatory purposes.