A multi-evidence approach in an Amazonian river based on land use, water quality, histopathological effects and habitat selection behavior in fish.

The Amazon rivers constitute the largest river basin in the world, with a high level of biodiversity. The Tocantins River is one of the most important rivers in this region, which has been impacted by different land uses. The objective of this study was to carry out a multi-evidence analysis focusing on the water quality of the Tocantins River, close to the municipality of Marabá-PA. We analyzed forest cover and water quality and, using the model organism Danio rerio, performed toxicity tests for histopathological effects, as well as the habitat selection approach by exposing fish to different river water samples in a multi-compartment device. The results showed that the studied area has already lost almost 30% of its forests in recent decades. Regarding water quality, the upstream (C1) and downstream (C5) points are the least impacted. On the other hand, the other points (C2-C4), closer to the city, greater input of pollutants was detected. Fish exposed to water samples from the most impacted sites showed several oedemas and hyperplastic cells in the gills. Regarding habitat selection behavior, there was a marked avoidance by samples with the highest contamination load. The results of this study lead to the understanding of the potential negative effects of human activities on local Amazonian biodiversity, since the potential toxicity of the environment, in conjunction with changes in the habitat selection process, could lead to a decline in populations of aquatic organisms, altering the environmental balance.

Ecological consequences when organisms avoid a contaminated environment: A study evaluating the toxicity of fipronil.

The ability of aquatic organisms to sense the surrounding environment chemically and interpret these signals correctly is crucial to their survival and ecological niche. This study applied the Heterogenous Multi-Habitat Assay System - HeMHAS to evaluate the avoidance potential of Daphnia magna to detect fipronil-contaminated habitats in a connected landscape after a short (48 h), previous, forced exposure to an environmentally relevant concentration of the same insecticide. The swimming of daphnids was also analyzed by recording the total distance covered. D. magna preferred areas with less contamination, although the effect of fipronil on their swimming ability (a decrease) was observed for all the concentrations tested. The application of non-forced multi-compartment exposure methodologies is a recent trend and is ecologically relevant as it is based on how contamination can really produce changes in an organism's habitat selection. Finally, we consider the importance of more non-forced exposure approaches where Stress Ecology can be aggregated to improve systemic understanding of the risk that contaminants pose to aquatic ecosystems from a broader landscape perspective.

Population and functional changes in a multispecies co-culture of marine microalgae and cyanobacteria under a combination of different salinity and temperature levels.

Changes in the temperature or salinity of ocean waters can affect marine organisms at multiple trophic levels. Both environmental variables could have an impact on marine microalgae populations. Therefore, the effect of the combination of three levels of temperature (20, 24 and 28 °C), and three levels of salinity (33, 36, and 39 PSU) were evaluated on the growth of a multispecies community of five common species of phytoplankton: (one cyanobacteria, Synechococcus sp., and four microalgae, Chaetoceros gracilis, Amphidinium carterae, Pleurochrysis roscoffensis and Rhodomonas baltica). The co-culture was monitored by flow cytometry under controlled conditions in a 96 h study. The effect of both variables on dissolved oxygen concentrations was measured using the SDR SensorDish Reader system. The results demonstrated that Synechococcus sp., C. gracilis, and A. carterae displayed a high growth at the temperature of 28 °C combined with the lowest salinity assayed. However, salinity increases negatively affected the growth of P. roscoffensis and R. baltica. Decreased salinity combined with decreased temperature exhibited a higher net O2 production. The interaction of two environmental factors related to global change such as temperature and salinity can cause structural (community growth) and functional (net oxygen production) changes in a phytoplanktonic community.

Behavioural, developmental and biochemical effects in zebrafish caused by ibuprofen, irgarol and terbuthylazine.

The increasing use of chemicals and their release into aquatic ecosystems are harming aquatic biota. Despite extensive ecotoxicological research, many environmental pollutants' ecological effects are still unknown. This study examined the spatial avoidance, behavioural and biochemical impacts of ibuprofen, irgarol, and terbuthylazine on the early life stages of zebrafish (Danio rerio) under a range of ecologically relevant concentrations (0-500 µg/L). Embryos were exposed following the OECD guideline "fish embryo toxicity test" complemented with biochemical assessment of AChE activity and behavioural analyses (swimming activity) using the video tracking system Zebrabox. Moreover, spatial avoidance was assessed by exposing 120 hpf-old larvae of D. rerio to a gradient of each chemical, by using the heterogeneous multi-habitat assay system (HeMHAS). The results obtained revealed that the 3 compounds delayed hatching at concentrations of 50 and 500 µg/L for both ibuprofen and irgarol and 500 µg/L for terbuthylazine. Moreover, all chemicals elicited a dose-dependent depression of movement (swimming distance) with LOEC values of 5, 500 and 50 µg/L for ibuprofen, irgarol and terbuthylazine, respectively. Zebrafish larvae avoided the three chemicals studied, with 4 h-AC50 values for ibuprofen, irgarol, and terbuthylazine of 64.32, 79.86, and 131.04 µg/L, respectively. The results of the HeMHAS assay suggest that larvae may early on avoid (just after 4 h of exposure) concentrations of the three chemicals that may later induce, apical and biochemical effects. Findings from this study make clear some advantages of using HeMHAS in ecotoxicology as it is: ecologically relevant (by simulating a chemically heterogeneous environmental scenario), sensitive (the perception of chemicals and the avoidance can occur at concentrations lower than those producing lethal or sublethal effects) and more humane and refined approach (organisms are not mandatorily exposed to concentrations that can produce individual toxicity).

Multi-level biological responses of Daphnia magna exposed to settleable atmospheric particulate matter from metallurgical industries.

Metallurgical industries are a continuous source of air pollution due to the amount of settleable particulate matter (SePM) they release. This SePM is a complex mixture formed by metallic nanoparticles and metals, which reach terrestrial and aquatic ecosystems and can be a significant source of contamination. The aim of this study was to evaluate the adverse effects of SePM at different levels of biological organization in order to estimate its ecological impacts on aquatic ecosystems. For this purpose, the crustacean Daphnia magna was exposed to different concentrations of SePM (0.01, 0.1, 1, 5, 10 g/L) using a multi-level response approach. The endpoints studied were: avoidance throughout 24 h in a non-forced exposure system, reproduction (total number of neonates per female after 21 days of exposure), acetylcholinesterase activity (AChE) after 48 h, and finally, the feeding rates during a short-term exposure (48 h) and a long-term exposure (21 day + 48 h). There was a negative effect of SePM on all responses measured at high concentrations. The avoidance was concentration-dependent and represented 88 % and 100 % at the two highest concentrations. The AChE activity was significantly inhibited at 5 and 10 g/L. The total number of neonates increased from 1 g/L of SePM and the first brood occurred earlier as of 5 g/L compared to control. The post-exposure feeding rates were lower during long-term exposure at the highest concentration. Chemical analyses were performed to characterize the metals present in this SePM, but this study did not report any direct relationship with toxicity, due to the chemical heterogeneity of the particles. The emission of compounds caused by anthropogenic activity may have significant ecological consequences, so it is important to consider these possible effects on aquatic biota generated by the mixture of metals present in SePM originated from metallurgical activities. Environmental and sectorial regulations are needed to prevent contamination and ecological disturbances.

Dissimilar behavioral and spatial avoidance responses by shrimps from tropical and temperate environments exposed to copper.

Behavioral changes associated with exposure to pollutants represent the earliest response for organisms confronted by perceivable chemical signals. This study was carried out with the objective of evaluating behavioral responses associated with different scenarios of exposure to pollutants (non-forced vs forced) in two shrimp species (Penaeus vannamei and Palaemon varians), representative of different latitudes and using copper as a model contaminant. The effects on locomotion were evaluated by exposing the shrimps to a range of copper concentrations (0, 0.5, 5, 50, and 250 µg/L) in the forced scenario. After exposure, the movement patterns for each shrimp were recorded and used to estimate changes in the shrimps' locomotion. For the non-forced scenario, the avoidance response was assessed by placing shrimps in a multi-compartment system where they were able to move freely along a gradient of copper (0, 0.5, 5, 50, and 250 µg/L). In terms of locomotion, an opposite trend was observed between the species: movements were significantly reduced in P. varians with concentrations above 50 µg/L, while hyperactivity was observed for P. vannamei. When exposed to a gradient of copper in the multi-compartment system, both species significantly avoided the highest concentrations of copper, although the repellence of copper was stronger for P. vannamei. In summary, both species of shrimps were able to recognize and avoid copper; however, in terms of locomotion, they showed an opposite behavioral reaction. These results show that a contamination event can have different behavioral outcomes depending on the species and complementing forced and non-forced exposure with species-specific information can be helpful to characterize and predict the effects of contaminants at higher biological levels.

Ecotoxicological assessment of the effects of fluoxetine on Daphnia magna based on acute toxicity, multigenerational reproduction effects, and attraction-repellence responses.

Fluoxetine, a common pharmaceutical used as an antidepressant, is already considered potentially hazardous to biota due to its increasing use and detection in European, North American, and Asian rivers. We studied the effects of fluoxetine on Daphnia magna, as we hypothesized that fluoxetine might have harmful effects, short and long-term, at different levels: survival, behaviour, and reproduction (offspring production). We applied two different approaches: (i) a scenario at environmentally relevant concentrations (0.1-1.0 µg/L) and (ii) a scenario simulating a future worsening of contamination (1-800 µg/L) until the reach of lethal concentrations. In the former, we examined whether there are multigenerational effects on reproduction and on the avoidance/colonisation behaviour in previously exposed populations. In the latter, three responses were assessed: survival, avoidance behaviour and reproduction. We did not detect differences in the reproduction output of D. magna among the treatments over the three generations examined. Irrespective of the multigenerational treatment, D. magna colonised the environments with fluoxetine in a similar way. In the second scenario, we determined the lethal concentration for 50% of the population (96 h-LC50 = 365 µg/L), which, in spite of the toxic effect, was attractive to organisms during the avoidance tests (24 h); in fact, D. magna were attracted (no repellence) even to the highest concentrations of fluoxetine tested (800 µg/L). Lastly, in a 21-day chronic toxicity test the reproduction output of D. magna increased with higher concentrations of fluoxetine. This effect might be related to the fact that the organisms in the contaminated treatment began their first reproduction earlier, when compared to that in the control treatments. In conclusion, this study discusses an identified hazard for aquatic biota due to the fluoxetine attraction effect and a predictive assessment of the consequences expected if its indiscriminate use increases.

Short and long-term exposure to the pesticides fipronil and 2,4-D: Effects on behavior and life history of Daphnia magna.

The high levels of contamination in aquatic ecosystems caused by pesticides and the organisms' consequent continuous exposure to it has made them vulnerable to damage. However, mobile organisms can avoid this continued exposure to contaminants by moving to less disturbed habitats. Therefore, through the use of the Heterogenous Multi-Habitat Assay System (HeMHAS), our objective was to evaluate the ability of Daphnia magna to detect and avoid habitats contaminated by fipronil and 2,4-D, in a spatially connected landscape. Further, the role of contamination by these pesticides, isolated and in mixtures, concerning the colonization of habitats by daphnids was also evaluated. Given that not all organisms successfully escape contamination, the chronic toxicity of the same pesticides using different parameters for D. magna (maternal survival, fecundity and maternal body length) was also evaluated. When evaluating the avoidance response by D. magna exposed to pesticides, there was no preference for the less contaminated areas for both compounds. However, organisms did not move to contaminated zones in the colonization experiments, with no immigration of daphnids to the zones with intermediate and the highest levels of fipronil, nor to the highest concentration of 2,4-D. Finally, the colonization by daphnids was significantly prevented when exposed to a mixture of the pesticides, in which the areas with the highest combinations of pesticide concentrations were not colonized by D. magna. Regarding the long-term chronic effects, negative consequences were observed, particularly for maternal body length, fecundity and maternal survival, due to the exposure to fipronil. Considering that pesticides can limit the areas colonized by organisms by making them unattractive, the risk of local population extinction may be underestimated if only standard endpoints involving forced exposure are studied.

Assessing the colonization by Daphnia magna of pesticide-disturbed habitats (chlorpyrifos, terbuthylazine and their mixtures) and the behavioral and neurotoxic effects.

The spread of pesticides in water bodies integrated into agricultural landscapes may prevent some areas from being colonized. In this study, the effects on the colonization responses of D. magna exerted by gradients of realistic environmental concentrations of the pesticides chlorpyrifos, terbuthylazine and their mixtures were tested in a novel multicompartment non-forced exposure system. Furthermore, the effects of both pesticides and their mixtures on the swimming behavior and the neurotransmission activity of D. magna were analyzed using a traditional forced exposure system. The synthesis and concentration of the main environmental metabolites of terbuthylazine were also analyzed. Results confirmed that D. magna exposed to mixture gradients were able to detect the pollutants and their colonization dynamics were drastically inhibited. The swimming behavior increased in D. magna exposed to the highest concentration of the mixture treatment. AChE activity was only significantly inhibited in the D. magna exposed to the highest concentration of chlorpyrifos. Changes in swimming behavior could not be directly related to the effects on AChE. Furthermore, the synthesis of the metabolite terbuthylazine 2-hydroxy during the course of the experiments was confirmed. These results demonstrate the importance of integrating pesticide mixtures in both non-forced and forced exposure systems during ecotoxicological assays.

Avoidance responses by Danio rerio reveal interactive effects of warming, pesticides and their mixtures.

Temperature variations and thermal extremes events caused by climate change can have profound implications for the toxicity of pesticides in aquatic organisms. Using an innovative system (Heterogeneous Multi-Habitat Test System - HeMHAS) that allows the simulation of different scenarios within a spatially heterogeneous landscape, the effects on the habitat selection of Danio rerio fish caused by the pesticides fipronil and 2,4-D were studied as single compounds and in mixture and integrated with air temperature variation (20, 24 and 28 °C). As a result, D. rerio detected and avoided both pesticides at air temperatures of 20 and 24 °C; however, at 28 °C no significant difference was observed in habitat choice by fish. Additionally, when pesticides were mixed in a heterogeneously contaminated landscape, it was observed that D. rerio detected contamination and preferred the clean zone at 20 and 24 °C; however, at 28 °C the potential to escape from the most contaminated areas was impaired. Thus, contamination by both pesticides made the habitat selection behavior of fish at 20 and 24 °C more noticeable. In addition, the association between pesticides and temperature showed negative effects on the response of fish to detect and escape from contaminated environments, suggesting the influence of temperature in altering the ability of the organism to provide an efficient response to stress.

Humane acute testing with tadpoles for risk assessment of chemicals: Avoidance instead of lethality.

In spite of the sensitivity of amphibians to contamination, data from fish have been commonly used to predict the effects of chemicals on aquatic life stages. However, recent studies have highlighted that toxicity data derived from fish species may not protect all the aquatic life stages of amphibians. For pesticide toxicity assessment (PTA), EFSA has highlighted that more information on lethal toxicity for the aquatic life stages of amphibians is still needed to reduce uncertainties. The current review aims to propose a test with amphibians based on spatial avoidance, as a more humane alternative method to the lethality tests for chemicals. A review of lethal toxicity tests carried out with amphibians in the period between 2018 and 2021 is presented, then we discuss the suitability of using fish toxicity data as a surrogate to predict the effects on more sensitive amphibian groups. The possible differences in sensitivity to chemicals may justify the need to develop further tests with amphibian embryos and larvae in order to reduce uncertainties. A new test is proposed focused on the avoidance behaviour of organisms fleeing from contamination to replace lethal tests. As avoidance indicates the threshold at which organisms will flee from contamination, a reduction in the population density, or its disappearance, at the local scale due to emigration is expected, with ecological consequences analogous to mortality. Avoidance tests provide an ethical advantage over lethal tests as they respect the concepts of the 3 Rs (mainly Refinement), reducing the suffering of the organisms.

Experimental evidence of contamination driven shrimp population dynamics: Susceptibility of populations to spatial isolation.

Contamination is likely to affect the composition of an ecological landscape, leading to the rupture of ecological connectivity among habitats (ecological fragmentation), which may impact on the distribution, persistence and abundance of populations. In the current study, different scenarios within a spatially heterogeneous landscape were simulated in the Heterogeneous Multi-Habitat Assay System (HeMHAS) to evaluate the potential effect that contamination (copper at 0.5 and 25 µg/L) might have on habitat selection by the estuarine shrimp Palaemon varians in combination with two other ecological factors: predator presence and food availability. As a result, P. varians detected and avoided copper; however, in the presence of the predation signal, shrimps shifted their response by moving to previously avoided regions, even if this resulted in a higher exposure to contamination. When encouraged to move towards environments with a high availability of food, a lower connectivity among the shrimp populations isolated by both contamination and predation risk simultaneously was evidenced, when compared to populations isolated only by the risk of predation. These results indicate that contamination might: (i) trigger avoidance in shrimps, (ii) prevent colonization of attractive foraging areas, (iii) enhance populations' isolation and (iv), make populations more susceptible to local extinction.

Can a mixture of agrochemicals (glyphosate, chlorpyrifos and chlorothalonil) mask the perception of an individual chemical? A hidden trap underlying ecological risk.

As aquatic environments associated with conventional agriculture are exposed to various pesticides, it is important to identify any possible interactions that modify their effects when in a mixture. We applied avoidance tests with Danio rerio, exposing juveniles to three relevant current use pesticides: chlorpyrifos (CPF), chlorothalonil (CTL) and glyphosate (Gly), individually and in binary mixtures (CPF-Gly and CTL-Gly). Our goal was to identify the potential of contaminants to trigger the avoidance response in fish and detect any changes to that response resulting from binary mixtures. Avoidance was assessed for three hours using an open gradient system with six levels of increasing concentrations. Fish avoided environmentally relevant concentrations of the three compounds. The avoidance of CPF [AC50 = 7.95 (3.3-36.3) µg/L] and CTL [AC50 = 3.41 (1.2-41.6) µg/L] was evident during the entire period of observation. In the case of Gly, the response changed throughout the experiment: initially (until 100 min) the fish tolerated higher concentrations of the herbicide [AC50 = 52.2 (12.1-2700) µg/L] while during the later period (after 100 min) a clearer avoidance [1.5 (0.8-4.2) µg/L] was observed. The avoidance recorded using CPF and CTL alone was attenuated by the presence of Gly. Applying an additive concentration model, Gly initially acted synergistically with the other two compounds, although this interaction was not observed during the later period. Avoidance gives us an idea of how the distribution of populations may be altered by contamination, our results suggest that in some mixtures this response may be inhibited, at least temporarily, thus masking the ecological risk of the exposure.

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.

Galaxolide and tonalide modulate neuroendocrine activity in marine species from two taxonomic groups.

Galaxolide (HHCB) and tonalide (AHTN) are polycyclic musk compounds (PMCs) used in household and personal care products that have been included on the list as emerging contaminants of environmental concern due to their ubiquity in aquatic and terrestrial environments. There still exists a dearth of information on the neurotoxicity and endocrine disrupting effects of these contaminants, especially for marine and estuarine species. Here, we assessed the neuroendocrine effects of HHCB and AHTN using adult clams, Ruditapes philippinarum, and yolk-sac larvae of sheepshead minnow, Cyprinodon variegatus. The clams were treated with concentrations (0.005-50 µg/L) of each compound for 21 days. Meanwhile, sheepshead minnow larvae were exposed to 0.5, 5 and 50 µg/L of HHCB and AHTN for 3 days. Enzyme activities related to neurotoxicity (acetylcholinesterase - AChE), neuroendocrine function (cyclooxygenase - COX), and energy reserves (total lipids - TL) were assessed in R. philippinarum. Gene expression levels of cyp19 and vtg1 were measured in C. variegatus using qPCR. Our results indicated induction of AChE and COX in the clams exposed to HHCB while AHTN exposure significantly inhibited AChE and COX. Gene expression of cyp19 and vtg1 in yolk-sac C. variegatus larvae exposed to 50 µg/L AHTN was significantly downregulated versus the control. The results of this study demonstrate that HHCB and AHTN might pose neurotoxic and endocrine disrupting effects in coastal ecosystems.

Fipronil and 2,4-D effects on tropical fish: Could avoidance response be explained by changes in swimming behavior and neurotransmission impairments?

Brazil is the largest producer of sugarcane, a crop largely dependent on chemical control for its maintenance. The insecticide fipronil and herbicide 2,4-D stand out among the most commonly used pesticides and, therefore, environmental consequences are a matter of concern. The present study aimed to investigate the toxicity mechanisms of Regent® 800 WG (a.i. fipronil) and DMA® 806 BR (a.i. 2,4-D) pesticides using forced and non-forced exposures through an integrative approach: firstly, to assess whether contamination by fipronil and 2,4-D can trigger the avoidance behavior of the fish Danio rerio (zebrafish) and Hyphessobrycon eques (serpae tetra or mato-grosso). Additionally, the effects on fish were analyzed considering the swimming behavior together with a biomarker of neurotoxicity, the activity of acetylcholinesterase (AChE). In avoidance tests with pesticide gradients, D. rerio avoided the highest concentrations of the two compounds and H. eques avoided only the highest concentration of 2,4-D. The swimming behavior (distance moved) was reduced and AChE was inhibited when D. rerio was exposed to fipronil. The 2,4-D affected the swimming (maximum speed) of H. eques, but AChE was not altered. Avoidance response seemed not to have been affected by possible effects of contaminants on swimming behavior and Ache activity. This study showed the importance of knowing the avoidance capacity, swimming behavior and neurotoxic effects of pesticides on fish in an integrated and realistic context of exposure in environments contaminated with pesticides and can be useful as ecologically relevant tools for ecological risk assessment.

The environmental risks of pharmaceuticals beyond traditional toxic effects: Chemical differences that can repel or entrap aquatic organisms.

The aim of the present study was to assess the risks of four different pharmaceutical active compounds (PhACs; diazepam, metformin, omeprazole and simvastatin). Acute and chronic toxicities were studied using the bacterium Aliivibrio fischeri and the microalgae Pseudokirchneriella subcapitata; while the repellency and attractiveness were assessed by avoidance tests with juvenile Cypirinus carpio using a multi-compartmented exposure system. Omeprazole was found to be an acutely toxic drug (EC50: 0.015 mg/L), while the other PhACs, except simvastatin, showed some chronic toxicity. Regarding avoidance, simvastatin and omeprazole induced an escape response for 50% of the fish population at 0.032 and 0.144 mg/L, respectively; contrarily, diazepam was attractive, even at lethal concentrations, representing a dangerous trap for organisms. The toxicity of the PhACs seemed not to be directly related to their repellency; and the mode of action seems to determine the repellency or attractiveness of the chemicals. Contamination by PhACs is of concern due to the environmental disturbance they might cause, either due to their acute and chronic toxicity (at the individual level), repellency (at the ecosystem level: loss of local biodiversity) or attraction to potentially lethal levels.

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.

Potential of environmental concentrations of the musks galaxolide and tonalide to induce oxidative stress and genotoxicity in the marine environment.

Polycyclic musk compounds have been identified in environmental matrices (water, sediment and air) and in biological tissues in the last decade, yet only minimal attention has been paid to their chronic toxicity in the marine environment. In the present research, the clams Ruditapes philippinarum were exposed to 0.005, 0.05, 0.5, 5 and 50 µg/L of the fragrances Galaxolide® (HHCB) and Tonalide® (AHTN) for 21 days. A battery of biomarkers related with xenobiotics biotransformation (EROD and GST), oxidative stress (GPx, GR and LPO) and genotoxicity (DNA damage) were measured in digestive gland tissues. HHCB and AHTN significantly (p < 0.05) induced EROD and GST enzymatic activities at environmental concentrations. Both fragrances also induced GPx activity. All concentrations of both compounds induced an increase of LPO and DNA damage on day 21. Although these substances have been reported as not acutely toxic, this study shows that they might induce oxidative stress and genotoxicity in marine organisms.