Transgenerational endocrine disruptor effects of cadmium in zebrafish and contribution of standing epigenetic variation to adaptation.

Evidence has emerged that environmentally-induced epigenetic changes can have long-lasting effects on gene transcription across generations. These recent findings highlight the need to investigate the transgenerational impacts of pollutants to assess their long term effects on populations. In this study, we investigated the transgenerational effect of cadmium on zebrafish across 4 generations. A first whole methylome approach carried out on fish of the first two generations led us to focus our investigations on the estradiol receptor alpha gene (esr1). We observed a sex-dependent transgenerational inheritance of Cd-induced DNA methylation changes up to the last generation. These changes were associated with single nucleotide polymorphisms (SNPs) that were themselves at the origin of the creation or deletion of methylation sites. Thus, Cd-induced genetic selection gave rise to DNA methylation changes. We also analyzed the transcription level of various sections of esr1 as well as estrogen responsive genes. While Cd triggered transgenerational disorders, Cd-induced epigenetic changes in esr1 contributed to the rapid transgenerational adaptation of fish to Cd. Our results provide insight into the processes underpinning rapid adaptation and highlight the need to maintain genetic diversity within natural populations to bolster the resilience of species faced with the global environmental changes.

Genetic and epigenetic interplay allows rapid transgenerational adaptation to metal pollution in zebrafish.

Despite still being a matter of debate, there is growing evidence that pollutant-induced epigenetic changes can be propagated across generations. Whereas such modifications could have long-lasting effects on organisms and even on population, environmentally relevant data from long-term exposure combined with follow-up through multiple generations remain scarce for non-mammalian species. We performed a transgenerational experiment comprising four successive generations of zebrafish. Only fish from the first generation were exposed to an environmentally realistic concentration of cadmium (Cd). Using a whole methylome analysis, we first identified the DNA regions that were differentially methylated in response to Cd exposure and common to fish of the first two generations. Among them, we then focused our investigations on the exon 3 (ex3) of the cep19 gene. We indeed recorded transgenerational growth disorders in Cd-exposed fish, and a mutation in this exon is known to cause morbid obesity in mammals. Its methylation level was thus determined in zebrafish from all the four generations by means of a targeted and base resolution method. We observed a transgenerational inheritance of Cd-induced DNA methylation changes up to the fourth generation. However, these changes were closely associated with genetic variations, mainly a single nucleotide polymorphism. This single nucleotide polymorphism was itself at the origin of the creation or deletion of a methylation site and deeply impacted the methylation level of neighboring methylation sites. Cd-induced epigenetic changes were associated with different mRNA transcripts and an improved condition of Cd fish. Our results emphasize a tight relationship between genetic and epigenetic mechanisms and suggest that their interplay and pre-existing diversity can allow rapid adaptation to anthropogenic environmental changes.

Metal contamination and heat stress impair swimming behavior and acetylcholinesterase activity in embryo-larval stages of the Mediterranean mussel, Mytilus galloprovincialis.

Behavioral parameters are increasingly considered sensitive and early bioindicators of toxicity in aquatic organisms. A video-tracking tool was specifically developed to monitor the swimming behaviour of D-larvae of the Mediterranean mussel, Mytilus galloprovincialis, in controlled laboratory conditions. Both maximum and average swimming speeds and trajectories were recorded. We then investigated the impact of copper and silver with or without a moderate rise of temperature on swimming behavior and acetylcholinesterase (AChE) activity of mussel D-larvae and the possible mechanistic link between both biological responses. Our results showed that copper and/or silver exposure, as well as temperature increase, disrupts the swimming behavior of mussel larvae which could compromise their dispersal and survival. In addition, the combined effect of temperature and metals significantly (p < 0.05) increased AChE activity in mussel larvae. Pearson's correlation analysis was performed and results showed that the AChE activity is positively correlated with maximum speeds (r = 0.71, p < 0.01). This study demonstrates the value of behavioral analyzes of aquatic invertebrates as a sensitive and integrate marker of the effects of stressors.

Effects of Biofilms and Particle Physical Properties on the Rising and Settling Velocities of Microplastic Fibers and Sheets.

Vertical dynamics of microplastics (MPs) in the water column are complex and not fully understood due to the diversity of environmental MPs and the impact of weathering and biofouling on their dynamical properties. In this study, we investigate the effects of the particle properties and biofilm on the vertical (settling or rising) velocity of microplastic sheets and fibers under laboratory conditions. The experiments focus on three types of MPs (polyester PES fibers, polyethylene terephthalate PET sheets, and polypropylene PP sheets) of nine sizes and two degrees of biological colonization. Even though pristine PES fibers and PET sheets had a similar density, the sinking velocity of fibers was much smaller and independent of their length. The settling or rising velocity of sheets increased with the particle size up to a threshold and then decreased in the wake of horizontal oscillations in large particles. Biofilms had unexpected effects on vertical velocities. Irregular biofilm distributions can trigger motion instabilities that decrease settling velocities of sheets despite the increase in density. Biofilms can also modify the orientation of fibers, which may increase their settling velocity. Finally, we selected the most performant theoretical formulation for each type of particle and proposed modifications to consider the effect of biofilm distribution.

Pesticide mixture toxicity assessment through in situ and laboratory approaches using embryo-larval stages of the pacific oyster (Magallana gigas).

Worsened state of oysters in French Arcachon Bay, demand an investigation of possible causes. This study evaluated the effects of an environmentally relevant mixture of five common pesticides on the early-life stages of the Pacific oyster (Magallana gigas). Laboratory assays with artificial mixture and in situ transplantation were complementarily used to investigate a series of sublethal endpoints. The laboratory exposure revealed developmental toxicity at 0.32 µg/L, which corresponds to mixture concentrations in Arcachon Bay. Downregulation of some gene transcriptions was observed at environmental level. No difference in larvae development was revealed among the three sites in Arcachon Bay. This study was the first to evaluate locomotion of oyster larvae exposed in situ. Suspected poor water quality in the inner part of Arcachon Bay was reflected by impairment at the molecular level. In conclusion, current concentrations of the tested pesticides in Arcachon Bay hinder larval development and affect several biological functions.

Parasitism in ecosystem engineer species: A key factor controlling marine ecosystem functioning.

Although parasites represent a substantial part of marine communities' biomass and diversity, their influence on ecosystem functioning, especially via the modification of host behaviour, remains largely unknown. Here, we explored the effects of the bopyrid ectoparasite Gyge branchialis on the engineering activities of the thalassinid crustacean Upogebia pusilla and the cascading effects on intertidal ecosystem processes (e.g. sediment bioturbation) and functions (e.g. nutrient regeneration). Laboratory experiments revealed that the overall activity level of parasitized mud shrimp is reduced by a factor 3.3 due to a decrease in time allocated to burrowing and ventilating activities (by factors 1.9 and 2.9, respectively). Decrease in activity level led to strong reductions of bioturbation rates and biogeochemical fluxes at the sediment-water interface. Given the world-wide distribution of mud shrimp and their key role in biogeochemical processes, parasite-mediated alteration of their engineering behaviour has undoubtedly broad ecological impacts on marine coastal systems functioning. Our results illustrate further the need to consider host-parasite interactions (including trait-mediated indirect effects) when assessing the contribution of species to ecosystem properties, functions and services.

Assessment of swimming behavior of the Pacific oyster D-larvae (Crassostrea gigas) following exposure to model pollutants.

This study describes an image analysis method that has been used to analyze the swimming behavior of native oyster D-larvae (Crassostrea gigas) from the Arcachon Bay (SW, France). In a second time, this study evaluated the impact of copper and S-metolachlor pollutants on D-larvae swimming activity and the possible relationship between developmental malformations and abnormal swimming behavior. Analyses in wild and cultivated oyster D-larvae were investigated during two breeding-seasons (2014 and 2015) at different sampling sites and dates. In controlled conditions, the average speed of larvae was 144 µm s-1 and the maximum speed was 297 µm s-1 while the trajectory is mainly rectilinear. In the presence of environmental concentration of copper or S-metolachlor, no significant difference in maximum or average larval speed was observed compared to the control condition but the percentage of circular trajectory increased significantly while the rectilinear swimming larvae significantly declined. The current study demonstrates that rectilinear trajectories are positively correlated to normal larvae while larvae with shell anomalies are positively correlated to circular trajectories. This abnormal behavior could affect the survival and spread of larvae, and consequently, the recruitment and colonization of new habitats.

Declining maerl vitality and habitat complexity across a dredging gradient: Insights from in situ sediment profile imagery (SPI).

Maerl beds form complex biogenic benthic habitats, characterized by high productivity as well as diverse biological communities. Disturbances associated with extraction and/or fishing activities using mobile bottom-contacting gears such as clam-dredges induce the most severe and long-term effects on these fragile habitats. We here investigated the effects of dredge-fishing on maerl in the bay of Brest (France). We quantified maerl beds structure and vitality across a fine scale quantified dredging intensity gradient through the acquisition of in-situ images of beds cross-section using Sediment Profile Imaging system (SPI). Declines in the proxies of maerl vitality and habitat complexity were measured across the gradient, and were associated with significant changes in the vertical distribution of live and dead maerl as well as of interstitial space. Fishing with dredges caused maerl mortality, substratum compaction, and decreasing habitat complexity. SPI imaging techniques also allowed for an assessment of changes in spatial heterogeneity that dredging created on several aspects of the structure and vitality of maerl beds. It suggests that direct and indirect disturbances induced by dredging are not acting at the same spatial scale, and can thereby differentially affect the ecosystem functions linked to vitality and habitat complexity.

Experimental Assessment of the Effects of Temperature and Food Availability on Particle Mixing by the Bivalve Abra alba Using New Image Analysis Techniques.

The effects of temperature and food addition on particle mixing in the deposit-feeding bivalve Abra alba were assessed using an experimental approach allowing for the tracking of individual fluorescent particle (luminophore) displacements. This allowed for the computations of vertical profiles of a set of parameters describing particle mixing. The frequency of luminophore displacements (jumps) was assessed through the measurement of both waiting times (i.e., the time lapses between two consecutive jumps of the same luminophore) and normalized numbers of jumps (i.e., the numbers of jumps detected in a given area divided by the number of luminophores in this area). Jump characteristics included the direction, duration and length of each jump. Particle tracking biodiffusion coefficients (Db) were also computed. Data originated from 32 experiments carried out under 4 combinations of 2 temperature (Te) and 2 food addition (Fo) levels. For each of these treatments, parameters were computed for 5 experimental durations (Ed). The effects of Se, Fo and Ed were assessed using PERmutational Multivariate ANalyses Of VAriance (PERMANOVAs) carried out on vertical depth profiles of each particle mixing parameter. Inversed waiting times significantly decreased with Ed whereas the normalized number of jumps did not, thereby suggesting that it constitutes a better proxy of jump frequency when assessing particle mixing based on the measure of individual particle displacements. Particle mixing was low during autumn temperature experiments and not affected by Fo, which was attributed to the dominant effect of low temperature. Conversely, particle mixing was high during summer temperature experiments and transitory inhibited by food addition. This last result is coherent with the functional responses (both in terms of activity and particle mixing) already measured for individual of the closely related clam A. ovata originating from temperate populations. It also partly resulted from a transitory switch between deposit- and suspension-feeding caused by the high concentration of suspended particulate organic matter immediately following food addition.

Long-term (1998-2010) large-scale comparison of the ecological quality status of gulf of lions (NW Mediterranean) benthic habitats.

A comprehensive Mediterranean data set has been used to address 3 questions associated with the use of sensitivity/tolerance based biotic indices to infer the Ecological Quality status (EcoQs) of benthic habitats. Our results showed: (1) a significant effect of the reference database on derived sensitivity/tolerance measure (ES500.05) as well as associated Benthic Quality Index values and derived EcoQs; (2) a lack of correlation neither between BQI and AZTI Marine Biotic Index values nor between BQI and Multivariate-AZTI Marine Biotic Index values; (3) a lack of correlation between the values of the Benthic Habitat Quality Index (index derived from Sediment Profile Imagery) and those of either of the 3 tested biotic indices; and (4) a general agreement between the 3 tested biotic indices in describing the lack of global trend for the EcoQs of the Gulf of Lions despite the occurrence of significant changes in benthic macrofauna composition between 1998 and 2010.

A bioturbation classification of European marine infaunal invertebrates.

Bioturbation, the biogenic modification of sediments through particle reworking and burrow ventilation, is a key mediator of many important geochemical processes in marine systems. In situ quantification of bioturbation can be achieved in a myriad of ways, requiring expert knowledge, technology, and resources not always available, and not feasible in some settings. Where dedicated research programmes do not exist, a practical alternative is the adoption of a trait-based approach to estimate community bioturbation potential (BPc). This index can be calculated from inventories of species, abundance and biomass data (routinely available for many systems), and a functional classification of organism traits associated with sediment mixing (less available). Presently, however, there is no agreed standard categorization for the reworking mode and mobility of benthic species. Based on information from the literature and expert opinion, we provide a functional classification for 1033 benthic invertebrate species from the northwest European continental shelf, as a tool to enable the standardized calculation of BPc in the region. Future uses of this classification table will increase the comparability and utility of large-scale assessments of ecosystem processes and functioning influenced by bioturbation (e.g., to support legislation). The key strengths, assumptions, and limitations of BPc as a metric are critically reviewed, offering guidelines for its calculation and application.