Personal care products: an emerging threat to the marine bivalve Ruditapes philippinarum.

In the last few decades, there has been a growing interest in understanding the behavior of personal care products (PCPs) in the aquatic environment. In this regard, the aim of this study is to estimate the accumulation and effects of four PCPs within the clam Ruditapes philippinarum. The PCPs selected were triclosan, OTNE, benzophenone-3, and octocrylene. A progressive uptake was observed and maximum concentrations in tissues were reached at the end of the exposure phase, up to levels of 0.68 µg g-1, 24 µg g-1, 0.81 µg g-1, and 1.52 µg g-1 for OTNE, BP-3, OC, and TCS, respectively. After the PCP post-exposure period, the removal percentages were higher than 65%. The estimated logarithm bioconcentration factor ranged from 3.34 to 2.93, in concordance with the lipophobicity of each substance. No lethal effects were found although significant changes were observed for ethoxyresorufin O-demethylase activity, glutathione S-transferase activity, lipid peroxidation, and DNA damage.

Are the BPA analogues an alternative to classical BPA? Comparison between 2D and alternative 3D in vitro neuron model to assess cytotoxic and genotoxic effects.

BPA is used in a wide range of consumer products with very concern toxicological properties. The European Union has restricted its use to protect human health. Industry has substituted BPA by BPA analogues. However, there is a lack of knowledge about their impacts. In this work, BPA and 5 BPA analogues (BPS, BPAP, BPAF, BPFL and BPC) have been studied in classical SH-SY5Y and the alternative 3D in vitro models after 24 and 96 h of exposure. Cell viability, percentage of ROS, cell cycle phases as well as the morphology of the spheroids were measured. The 2D model was more sensitive than the 3D models with differences in cell viability higher than 60% after 24 h of exposure, and different mechanisms of ROS production. After chronic exposure, both models were more affected in comparison to the 24 h exposure. After a recovery time (96 h), the spheroids exposed to 2.5-40 µM were able to recover cell viability and the morphology. Among the BPs tested, BPFL>BPAF>BPAP and >BPC revealed higher toxicological effects, while BPS was the only one with lower effects than BPA. To conclude, the SH-SY5Y 3D model is a suitable candidate to perform more reliable in vitro neurotoxicity tests.

Single and multispecies microalgae toxicological tests assessing the impact of several BPA analogues used by industry.

BPA is a hazard for human and environmental health and recently BPA was added to the Candidate List of substances of very high concern by European Chemical Agency (ECHA). In accordance with this proposal, the authorities have encouraged the replacement of BPA by BPA analogues; however, little is known about the impact of these compounds on the environment. Due to this situation five BPA analogues (BPS, BPAP, BPAF, BPFL and BPC) were chosen in order to study their effects on marine primary producers. Three marine microalgae species (Phaeodactylum tricornutum, Tetraselmis suecica and Nannochloropsis gaditana) were selected for single and multispecies tests concerning the ecotoxicological effects of these BPA analogues. Microalgae were exposed to BPs over 72 h at different dosages (5, 20, 40, 80, 150 and 300 µM). Responses such as: growth, ROS production, cell complexity, cell size, autofluorescence of chlorophyll a, effective quantum yield of PSII and pigment concentrations were assessed at 24, 48 and 72 h. The results revealed that BPS and BPA showed lower toxicity to microalgae in comparison with BPFL > BPAF > BPAP and >BPC for the endpoints studied. N. gaditana was the least sensitive microalgae in comparison to P. tricornutum and T. suecica. However, a different trend was found in the multispecies tests where T. suecica dominated the microalgae community in relation to N. gaditana and P. tricornutum. The results of this work revealed for first time that present day BPA analogues are a threat and not a safe substitute for BPA in terms of the marine phytoplanktonic community. Therefore, the results of their impact on aquatic organisms should be shared.

Contamination from microplastics and other anthropogenic particles in the digestive tracts of the commercial species Engraulis encrasicolus and Sardina pilchardus.

Fragments of microplastics (<5 mm) found in commercial species of fish, crustaceans, and bivalves, are an issue of global concern. The bioaccumulation of microplastics and other anthropogenic particles in different levels of the food web may provoke unwanted impacts on marine ecosystems and cause pernicious effects on human health. Here, we study the presence of anthropogenic particles and the fraction of microplastics in the target organs of two representative commercial fish species in Spain; the European anchovy (Engraulis encrasicolus) and the European pilchard (Sardina pilchardus). The individuals were sampled along the continental shelf of the Gulf of Cádiz, from the Bay of Cádiz to Cape Santa Maria. The isolation of the microplastics (MPs) was carried out with a complete alkaline-oxidant organic digestion (KOH-H2O2) of the digestive tract, including both the contents ingested and the muscle tissues. Anthropogenic particles were found in all individuals of both species with an average of 8.94 ± 5.11 items·ind-1. Fibres made up 93 % of the items while fragments and films were represented by the remaining 7 %. The average size of the anthropogenic particles was 0.89 ± 0.82 mm. In addition to the fragment and film particles identified as microplastics, 29 % of the fibres were estimated to be microplastics by Fourier-transform infrared spectroscopy (FTIR) analysis. The main polymer found in both species was nylon. No significant correlation was found between the abundance and size of anthropogenic particles ingested and individual size or other body variables. The analysis of similarities (ANOSIM) and the distanced-based multiple linear regression model showed a high homogeneity in anthropogenic particle contamination in both species throughout the study area along the continental shelf of the Gulf of Cádiz.

Adverse (geno)toxic effects of bisphenol A and its analogues in hepatic 3D cell model.

Bisphenol A (BPA) is one of the most widely used and versatile chemical compounds in polymer additives and epoxy resins for manufacturing a range of products for human applications. It is known as endocrine disruptor, however, there is growing evidence that it is genotoxic. Because of its adverse effects, the European Union has restricted its use to protect human health and the environment. As a result, the industry has begun developing BPA analogues, but there are not yet sufficient toxicity data to claim that they are safe. We investigated the adverse toxic effects of BPA and its analogues (BPS, BPAP, BPAF, BPFL, and BPC) with emphasis on their cytotoxic and genotoxic activities after short (24-h) and prolonged (96-h) exposure in in vitro hepatic three-dimensional cell model developed from HepG2 cells. The results showed that BPFL and BPC (formed by an additional ring system) were the most cytotoxic analogues that affected cell viability, spheroid surface area and morphology, cell proliferation, and apoptotic cell death. BPA, BPAP, and BPAF induced DNA double-strand break formation (γH2AX assay), whereas BPAF and BPC increased the percentage of p-H3-positive cells, indicating their aneugenic activity. All BPs induced DNA single-strand break formation (comet assay), with BPAP (≥0.1 µM) being the most effective and BPA and BPC the least effective (≥1 µM) under conditions applied. The results indicate that not all of the analogues studied are safer alternatives to BPA and thus more in-depth research is urgently needed to adequately evaluate the risks of BPA analogues and assess their safety for humans.

Products released from surgical face masks can provoke cytotoxicity in the marine diatom Phaeodactylum tricornutum.

Surgical face masks are more present than ever as personal protective equipment due to the COVID-19 pandemic. In this work, we show that the contents of regular surgical masks: i) polypropylene microfibres and ii) some added metals such as: Al, Fe, Cu, Mn, Zn and Ba, may be toxic to some marine life. This work has got two objectives: i) to study the release rate of the products from face masks in marine water and ii) to assess the toxicity in Phaeodactylum tricornutum of these by-products. To achieve these two objectives, we performed release kinetic experiments by adding masks in different stages of fragmentation to marine water (i.e. whole face masks and fragments of them 1.52 ± 0.86 mm). Released microfibres were found after one month in shaking marine water; 0.33 ± 0.24 and 21.13 ± 13.19 fibres·mL-1 were collected from the whole and fragmented face masks, respectively. Significant amounts of dissolved metals such as Mn, Zn and Ni, as well as functional groups only in the water containing the face mask fragments were detected. Water from both treatments was employed to study its toxicity on the marine diatom. Only the water from the face mask fragments showed a significant, dose-dependent, decrease in cell density in P. tricornutum; 53.09 % lower than in the controls. Although the water from the face mask fragments showed greater effects on the microalgae population than the water from the whole face mask, the latter treatment did show significant changes in the photosynthetic apparatus and intrinsic properties of the cells. These results indicate that during fragmentation and degradation face masks a significant chemical print can be observed in the marine environment.

Coastal gradients of small microplastics and associated pollutants influenced by estuarine sources.

Small microplastics (SMPs) in the gulf of Cadiz was sampled at 5 m depth by pumping it through the ship's pipe system and filtered through a 45 µm mesh size net. Our study reveals that higher densities have been found (130 mg·m-3) compared to other regions worldwide and these densities decreased from the coastline to the outer stations, showing a general coastal gradient influenced by estuarine outflows. SMPs with a size range between 45 and 193 µm were predominant and most of them composed by polyethylene and polypropylene. The metals associated with the MPs were mainly Na (21.1%), K (11.3%), Fe (8.5%), Ca (2.1%), Cr (1.8%), Zr (13.3%) and Hf (0.7%). The high proportion of Zr compared to Fe, which is different from what can be found in the environment, suggests that this metal is intrinsic to the materials used in catalytic processes during plastic production.

Surgical face masks as a source of emergent pollutants in aquatic systems: Analysis of their degradation product effects in Danio rerio through RNA-Seq.

Surgical face masks are the most popularised and effective personal equipment for protecting public health during the COVID-19 pandemic. They are composed of plastic polymer fibres with a large amount of inorganic and organic compounds that can be released into aquatic environments through degradation processes. This source of microplastics and inorganic and organic substances could potentially impact aquatic organisms. In this study, the toxicogenomic effects of face masks at different stages of degradation in water were analysed in zebrafish larvae (Danio rerio) through RNA-Seq. Larvae were exposed for 10 days to three treatments: 1) face mask fragments in an initial stage of degradation (poorly degraded masks -PDM- products) with the corresponding water; 2) face mask fragments in an advanced stage of degradation (highly degraded masks -HDM- products) with the corresponding water; and 3) water derived from HDM (W-HDM). Transcriptome analyses revealed that the three treatments provoked the down-regulation of genes related to reproduction, especially the HDM products, suggesting that degradation products derived from face masks could act as endocrine disruptors. The affected genes are involved in different steps of reproduction, including gametogenesis, sperm-egg recognition and binding or fertilisation. Immune-related genes and metabolic processes were also differentially affected by the treatments.

Are habitable clean areas in heterogeneously contaminated landscapes functioning as escape zones for fish populations to alleviate stress?

Environmental contamination is a problem that reduces the quality of ecosystems and may make them unsuitable to accommodate life. As many ecosystems are connected, some organisms avoid the stress from continuous exposure to contaminants by moving towards less disturbed areas. However, the landscapes in which organisms move might vary regarding the concentrations of contaminants, in the form of gradients or patches of contamination. Therefore, although it is expected that organisms prefer clean areas, their sporadic contact with contamination should not be ignored, as the greater the probability of being in contact with contaminated areas, the higher the stress. The aim of this study was to assess how the stress (cortisol levels) of zebrafish (Danio rerio) varies as a consequence of heterogeneity in the chemical composition of the habitats and the presence of uncontaminated areas in this heterogeneous landscape. Zebrafish were exposed to heterogeneous contamination scenarios containing different concentrations of copper along a free-choice multi-compartmented system, in which they were able to flee from the most contaminated areas. Fish escaped from the most contaminated areas with an avoidance by 50% of population (AC50) at concentrations of 41 (copper gradient scenario), 25 (spatially limited contamination scenario) and 69 (highly contaminated scenario) µg/L. Higher cortisol levels were observed in the populations exposed to homogeneously contaminated and highly contaminated (by copper) scenarios (both with no acceptable clean area to flee to). In summary, the uncontaminated areas might be crucial for the spatial dynamics of fish populations in a chemically heterogeneous landscape due to their role as escape zones to alleviate stress.

Could Contamination Avoidance Be an Endpoint That Protects the Environment? An Overview on How Species Respond to Copper, Glyphosate, and Silver Nanoparticles.

The use of non-forced multi-compartmented exposure systems has gained importance in the assessment of the contamination-driven spatial avoidance response. This new paradigm of exposure makes it possible to assess how contaminants fragment habitats, interfering in the spatial distribution and species' habitat selection processes. In this approach, organisms are exposed to a chemically heterogeneous scenario (a gradient or patches of contamination) and the response is focused on identifying the contamination levels considered aversive for organisms. Despite the interesting results that have been recently published, the use of this approach in ecotoxicological risk studies is still incipient. The current review aims to show the sensitivity of spatial avoidance in non-forced exposure systems in comparison with the traditional endpoints used in ecotoxicology under forced exposure. To do this, we have used the sensitivity profile by biological groups (SPBG) to offer an overview of the highly sensitive biological groups and the species sensitive distribution (SSD) to estimate the hazard concentration for 5% of the species (HC5). Three chemically different compounds were selected for this review: copper, glyphosate, and Ag-NPs. The results show that contamination-driven spatial avoidance is a very sensitive endpoint that could be integrated as a complementary tool to ecotoxicological studies in order to provide an overview of the level of repellence of contaminants. This repellence is a clear example of how contamination might fragment ecosystems, prevent connectivity among populations and condition the distribution of biodiversity.

An integrative toxicogenomic analysis of plastic additives.

In developed countries, contact with plastics is constant. Plastics contain a vast number of additives such as plasticisers, stabilisers, antioxidants, flame retardants, etc., that can impact human health. Most of them have been studied separately; however, an integrative approach to identify genes, biological processes, molecular functions, and diseases linked to exposure to these compounds has not been addressed until now. The genes most commonly affected by plastic additives are related to apoptosis, cell death, proliferation and differentiation, immunity and insulin-related processes, and are mainly associated with cancer, mental disorders, diabetes mellitus type II and obesity. The most commonly affected molecular functions included steroid hormone receptor activity implicated in cancer, mental disorders, immune signalling and gonadotropin-releasing hormones. These processes and functions affected by plastic additives are related to the diseases of the developed world, most of which are linked to the endocrine system, such as cancer, diabetes, infertility and obesity. The strong interconnection among the top 50 genes modulated by plastic additives shows that the pathways affected are strongly interrelated. Therefore, studying the effects of plastic additives through a single-compound approach cannot be sufficient and a holistic approach is more appropriate for evaluating the potential effects of plastics in human health.

Effects of classical PKC activation on hippocampal neurogenesis and cognitive performance: mechanism of action.

Hippocampal neurogenesis has widely been linked to memory and learning performance. New neurons generated from neural stem cells (NSC) within the dentate gyrus of the hippocampus (DG) integrate in hippocampal circuitry participating in memory tasks. Several neurological and neuropsychiatric disorders show cognitive impairment together with a reduction in DG neurogenesis. Growth factors secreted within the DG promote neurogenesis. Protein kinases of the protein kinase C (PKC) family facilitate the release of several of these growth factors, highlighting the role of PKC isozymes as key target molecules for the development of drugs that induce hippocampal neurogenesis. PKC activating diterpenes have been shown to facilitate NSC proliferation in neurogenic niches when injected intracerebroventricularly. We show in here that long-term administration of diterpene ER272 promotes neurogenesis in the subventricular zone and in the DG of mice, affecting neuroblasts differentiation and neuronal maturation. A concomitant improvement in learning and spatial memory tasks performance can be observed. Insights into the mechanism of action reveal that this compound facilitates classical PKCα activation and promotes transforming growth factor alpha (TGFα) and, to a lesser extent, neuregulin release. Our results highlight the role of this molecule in the development of pharmacological drugs to treat neurological and neuropsychiatric disorders associated with memory loss and a deficient neurogenesis.

Not Only Toxic but Repellent: What Can Organisms' Responses Tell Us about Contamination and What Are the Ecological Consequences When They Flee from an Environment?

The ability of aquatic organisms to sense the surrounding environment chemically and interpret such signals correctly is crucial for their ecological niche and survival. Although it is an oversimplification of the ecological interactions, we could consider that a significant part of the decisions taken by organisms are, to some extent, chemically driven. Accordingly, chemical contamination might interfere in the way organisms behave and interact with the environment. Just as any environmental factor, contamination can make a habitat less attractive or even unsuitable to accommodate life, conditioning to some degree the decision of organisms to stay in, or move from, an ecosystem. If we consider that contamination is not always spatially homogeneous and that many organisms can avoid it, the ability of contaminants to repel organisms should also be of concern. Thus, in this critical review, we have discussed the dual role of contamination: toxicity (disruption of the physiological and behavioral homeostasis) vs. repellency (contamination-driven changes in spatial distribution/habitat selection). The discussion is centered on methodologies (forced exposure against non-forced multi-compartmented exposure systems) and conceptual improvements (individual stress due to the toxic effects caused by a continuous exposure against contamination-driven spatial distribution). Finally, we propose an approach in which Stress and Landscape Ecology could be integrated with each other to improve our understanding of the threat contaminants represent to aquatic ecosystems.

Immunotoxicity of polystyrene nanoplastics in different hemocyte subpopulations of Mytilus galloprovincialis.

Plastic represents 60-80% of litter in the ocean. Degradation of plastic to small fragments leads to the formation of microplastics (MPs <5 mm) and nanoplastics (NPs <1 µm). One of the most widely used and representative plastics found in the ocean is polystyrene (PS). Among marine organisms, the immune system of bivalves is recognized as suitable to assess nanomaterial toxicity. Hemocyte subpopulations [R1 (large granular cells), R2 (small semi-granular cells) and R3 (small agranular or hyaline cells)] of Mytilus galloprovincialis are specialized in particular tasks and functions. The authors propose to examine the effects of different sizes (50 nm, 100 nm and 1 µm) PS NPs on the different immune cells of mussels when they were exposed to (1 and 10 mg·L-1) of PS NPs. The most noteworthy results found in this work are: (i) 1 µm PS NPs provoked higher immunological responses with respect to 50 and 100 nm PS NPs, possibly related to the higher stability in size and shape in hemolymph serum, (ii) the R1 subpopulation was the most affected with respect to R2 and R3 concerning immunological responses and (iii) an increase in the release of toxic radicals, apoptotic signals, tracking of lysosomes and a decrease in phagocytic activity was found in R1.

Ingestion and bioaccumulation of polystyrene nanoplastics and their effects on the microalgal feeding of Artemia franciscana.

Nanoplastics (NPs) have become one of the most serious environmental problems nowadays. The environmental issues linked to NPs are attributed to the effects after ingestion in marine organisms. Due to the incipient and controversial information about the effects of PS NPs on the feeding of organisms, the aim of this work is to assess (i) digestion dynamics of Artemia franciscana when exposed to PS NPs as the lowest concentration of PS NPs reported in toxicity test [0 (control), 0.006 and 0.6 mg·L-1] and possible interferences in the ingestion of microalgae and (ii) the accumulation and depuration of PS NPs by A. franciscana. Artemia were subjected to ingestion experiments [24 h and 3.5 h], in which the organisms were exposed to PS NPs or to PS NPs + microalgae. Post-exposure feeding (24 h exposure and 2 h feeding) and depuration (24 h exposure and 24 h of depuration) were also carried out. More than 90% of the PS NPs were ingested by Artemia and bioaccumulated in the mandible, stomach, gut, tail gut and appendages after 24 h. The ingestion of microalgae was not affected by the presence of the PS NPs. Data of post-exposure feeding indicated that Artemia previously exposed to plastic and/or microalgae presented similar microalgal ingestion (around 70%); the highest microalgal consumption (around 90%) was recorded in the treatment in which Artemia were previously starved (no plastic and no microalgae). The presence of PS NPs in the gut after the depuration experiments indicates that 24 h was not enough to eliminate the PS NPs.

Microplastics do not affect standard ecotoxicological endpoints in marine unicellular organisms.

In this study, the acute toxicity of microplastics (MPs) on unicellular organisms as marine decomposers and microalgae was assessed, by evaluating standards endpoints included in International Standard Organization (ISO) protocols. The bacteria Vibrio fischeri and the diatom Phaeodactylum tricornutum were exposed to different sizes (1-500 µm) of polyethylene MPs in order to evaluate bioluminescence inhibition and microalgal growth. No acute toxicity was found on bacteria or microalgae in an order of magnitude above environmentally relevant concentrations, suggesting that tested MPs did not affect the investigated biological processes. In conclusion, standard ecotoxicological endpoints are not sufficiently sensitive to assess the potential effects of MPs on decomposers and primary producers, conversely to nanoplastics. These findings highlight that the current approach for MP risk assessment in unicellular species should be revised, by providing alternative endpoints to be included in standardized protocols, able to monitor the fate and biological effects of MPs.

Biochemical response of the clam Ruditapes philippinarum to silver (AgD and AgNPs) exposure and application of an integrated biomarker response approach.

Silver is a ubiquitous metal in the marine environment which can be accumulated by marine organisms. In order to assess the effect of dissolved silver (AgD) and AgNPs in R. philippinarum, the organisms were exposed to 20 µg L-1 of AgD and AgNPs (15 nm) over 7 days. Bioaccumulation of the metal and oxidative and detoxification biomarkers were studied in control and exposed clams. Ag was accumulated in gills and digestive glands. Results for biochemical biomarkers (superoxide dismutase, catalase and glutathione reductase activity, lipid peroxidation and metallothionein provoked a general increase in the integrated biomarker response index (IBR) values) indicating the induction of oxidative stress in the clams exposed to both Ag treatments. Therefore, the presence of Ag forms at the tested concentration in the aquatic medium represent a risk for R. philippinarum.

Genomics and immunity of the Mediterranean mussel Mytilus galloprovincialis in a changing environment.

The Mediterranean mussel (Mytilus galloprovincialis) is a marine invasive species cultured all over the world. Mussels are an appreciated resource in local aquaculture enterprises because of their robust production and resilience that translates into a reliable economic value. So far, no massive mortalities have been reported in natural or cultured populations of this species. In the last years, the knowledge about its immune system has greatly improved but there are still many questions to be answered. One of them is why mussels, with their high filtering activity, are able to be exposed to a high number of potential pathogens without getting infected and without developing an elevated inflammatory response. The sequencing of the mussel genome has revealed a very complex organization with high heterozygosity, abundance of repetitive sequences and extreme intraspecific sequence diversity among individuals, mainly in immune related genes. Among those genes, antimicrobial peptides are the most expressed gene families in mussels, highly polymorphic and with antimicrobial effect against molluscs pathogens, but also against pathogens of lower vertebrates and humans. The combination of a complex genome with the adaptation of mussel immune system to a changing environment could explain this high variability, not only in immune-related genes, but also in the functional response among individuals sampled in the same location and date.

Are the primary characteristics of polystyrene nanoplastics responsible for toxicity and ad/absorption in the marine diatom Phaeodactylum tricornutum?

Nowadays, the occurrence of a large volume of plastic litter in oceanic and coastal zones has increased concern about its impacts on marine organisms. The degradation of plastic polymers leads to the formation of smaller fragments at both micro and nano scale (<5 mm and <1 µm respectively). Nanoplastics (NPs), due to their smaller size and high specific surface area can establish colloidal interactions with marine microalgae, therefore potential toxicity can be led. . To assess this hypothesis, the aim of the present study is to examine the behaviour of polystyrene nanoparticles (PS NPs) of different sizes (50 and 100 nm) in marine water and their possible effects at different physiological and cellular levels in the marine diatom Phaeodactylum tricornutum. Different biomarkers and stress responses in P. tricornutum were analysed when organisms were exposed to environmentally relevant PS NPs concentrations between 0.1 and 50 mg L-1. Our results showed significant differences between controls and exposure microalgae, indicating toxicity. After 24 h, an increase in oxidative stress biomarkers, damage to the photosynthetic apparatus, DNA damage and depolarization of mitochondrial and cell membrane from 5 mg L-1 were observed. Further after 72 h the inhibition of population growth and chlorophyll content were observed. Examining effects the effects related to PS NPs size, the smallest (50 nm) induced greater effects at 24 h while bigger PS NPs (100 nm) at72 h. This bigger particles (100 nm) showed more stability (in size distribution and spherical form) in the different culture media assayed, when compared with the rest of particles used. Strong adsorption and/or internalization of PS NPs was confirmed through changes in cell complexity and cell size as well as the fluorescence of 100 nm fluoresbrite PS NPs after washing cell surface.

Erythromycin sensitivity across different taxa of marine phytoplankton. A novel approach to sensitivity of microalgae and the evolutionary history of the 23S gene.

Erythromycin has been recorded in coastal waters and could pose a severe threat to marine microbial life. Macrolides such as erythromycin may affect microalgae by inhibiting the pathways involved in protein synthesis. Toxicological testing of microalgae has proven to be a useful tool for the risk assessment of a substance affecting phytoplankton. Due to the controversial results concerning the sensitivity of microalgal species to erythromycin found in the literature, the goals of this work were, initially, to assess the erythromycin sensitivity of different species of marine microalgae from different and representative taxonomic groups; and, secondly, to examine whether the sensitivity to erythromycin could be explained by the differences in the phylogenetic evolution. We chose eight species: two green algae, four heterokonts, one haptophyte and one dinoflagellate, which were then exposed to erythromycin (0.1 to 10 mg L-1). Our results showed a wide range of sensitivities indicating that the biology of each species was primarily responsible for the variation observed. To test the second objective, we contrasted different ecotoxicological endpoints (growth, cellular properties and status of the photosynthetic apparatus) with the phylogenetic distribution [eukaryotic host (concatenated nuclear tree), evolutionary history of the chloroplast (16S tree), efficiency and repair of photosystem II (psbA tree), and the binding site of erythromycin (23S tree)] of the species. We found that the growth inhibition of microalgae as a toxicological endpoint was the endpoint best explained by the topology of the 23S rRNA gene tree when it was modelled following a non-stationary evolutionary process.