How do life-history traits influence the fate of intertidal and subtidal Mytilus galloprovincialis in a changing climate?

Coastal organisms (i.e. intertidal or upper subtidal species) live in between the terrestrial and aquatic realms, making them particularly vulnerable to climate change. In this context, intertidal organisms may suffer from the predicted sea level rise (increasing their submerged time) while subtidal organisms may suffer from anthropically-induced hypoxia and its consequences. Although there is some knowledge on how coastal organisms adapt to environmental changes, the biochemical and physiological consequences of prolonged submergence periods have not yet been well characterized. Thus, the present study aimed to assess the biochemical alterations experienced by intertidal organisms maintained always under tidal exposure (IT); intertidal organisms maintained submersed (IS); subtidal organisms maintained always submersed (SS); subtidal organisms under tidal exposure (ST). For this, Mytilus galloprovincialis specimens from contiguous intertidal and subtidal populations were exposed to the above mentioned conditions for twenty-eight days. Results indicated that both intertidal and subtidal mussels are adapted to the oxidative stress pressure caused by tidal and submerged conditions tested. Intertidal mussels did not seem to be negatively affected by submergence while ST specimens were energetically challenged by tidal exposure. Both IT and ST mussels consumed glycogen to fuel up mechanisms aiming to maintain redox homeostasis. Overall, both intertidal and subtidal populations were capable of coping with tidal exposure, although the strategies employed differed between them. These findings indicate that although IT mussels may not significantly suffer from the longer-term submergence, hypoxic events occurring in the context of global warming and other anthropogenic impacts may have consequences on both IT and ST populations. Altogether, it is important to highlight that tides may act as a confounding factor in experiments concerning coastal organisms, as it causes additional physiological and biochemical perturbations.

Will temperature rise change the biochemical alterations induced in Mytilus galloprovincialis by cerium oxide nanoparticles and mercury?

It is known that, for marine coastal ecosystems, pollution and global warming are among the most threatening factors. Among emerging pollutants, nanoparticles (NPs) deserve particular attention as their possible adverse effects are significantly influenced by environmental factors such as salinity, pH and temperature, as well as by their ability to interact with other contaminants. In this framework, the present study aimed to evaluate the potential interactions between CeO2 NPs and the toxic classic metal mercury (Hg), under current and warming conditions. The marine bivalve Mytilus galloprovincialis was used as biological model and exposed to CeO2 NPs and Hg, either alone or in combination, for 28 day at 17 °C and 22 °C. A suite of biomarkers related to energetic metabolism, oxidative stress/damage, redox balance, and neurotoxicity was applied in exposed and non-exposed (control) mussels. The Hg and Ce accumulation was also assessed. Results showed that the exposure to CeO2 NPs alone did not induce toxic effects in M. galloprovincialis. On the contrary, Hg exposure determined a significant loss of energetic metabolism and a general impairment in biochemical performances. Hg accumulation in mussels was not modified by the presence of CeO2 NPs, while the biochemical alterations induced by Hg alone were partially canceled upon co-exposure with CeO2 NPs. The temperature increase induced loss of metabolic and biochemical functions and the effects of temperature prevailed on mussels exposed to pollutants acting alone or combined.

Complex interactions of rare earth elements in aquatic systems: Comparing observed and predicted cellular responses on Mytilus galloprovincialis.

Recent societal and technological developments have led to new sources of contamination, particularly from electronic waste (e-waste). The rapid increase in e-waste, combined with inadequate disposal and recycling practices has resulted in rising levels of hazardous substances in aquatic systems, including rare-earth elements (REEs). However, the effects of REEs on aquatic organisms remain poorly understood. This lack of understanding is concerning since REEs can simultaneously appear in aquatic systems. Thus, this study aimed to evaluate the impacts of Yttrium (Y), Lanthanum (La), and Gadolinium (Gd), individually and as mixtures on the mussel species Mytilus galloprovincialis. Biomarkers related to metabolism, energy reserves, defence enzymes, redox balance, cellular damage, and neurotoxicity were analyzed. The results obtained showed that Y alone caused minimal stress, while Gd, La, and their mixtures induced from moderate to severe stress, increasing metabolic activity, and enzyme responses. This study highlights the ecological impacts of REEs mixtures on aquatic organisms. The complex interactions and additive effects, especially with Gd, underline the need for further research on contaminant mixtures.

How predicted temperature and salinity changes will modulate the impacts induced by terbium in bivalves?

The threat of climate change, which includes shifts in salinity and temperature, has generated a global concern for marine organisms. These changes directly impact them and may alter their susceptibility to contaminants, such as terbium (Tb), found in electronic waste. This study assessed how decreased and increased salinity, as well as increased temperature, modulates Tb effects in Mytilus galloprovincialis mussels. After an exposure period of 28 days, Tb bioaccumulation and biochemical changes were evaluated. Results indicated no significant modulation of salinity and temperature on Tb accumulation, suggesting detoxification mechanisms and adaptations. Further analysis showed that Tb exposure alone caused antioxidant inhibition and neurotoxicity. When exposed to decreased salinity, these Tb-exposed organisms activated defense mechanisms, a response indicative of osmotic stress. Moreover, increased salinity also led to increased oxidative stress and metabolic activity in Tb-exposed organisms. Additionally, Tb-exposed organisms responded to elevated temperature with altered biochemical activities indicative of damage and stress response. Such responses suggested that Tb effects were masked by osmotic and heat stress. This study provides valuable insights into the interactions between temperature, salinity, and contaminants such as Tb, impacting marine organisms. Understanding these relationships is crucial for mitigating climate change and electronic waste effects on marine ecosystems.

Yttrium effects on the Mediterranean mussel under a scenario of salinity shifts and increased temperature.

Climate change (CC) induces significant worldwide alterations in salinity and temperature, impacting ecosystems and their services. Marine organisms, susceptible to these changes, may experience modified vulnerability to anthropogenic contaminants, including rare-earth elements (REEs) such as yttrium (Y) derived from electronic waste. This study investigated the influence of temperature and salinity changes on the impacts of Y in Mytilus galloprovincialis mussels. Organisms were subjected to Y (0 and 10 µg/L) for 28 days under three salinity scenarios (20, 30 (control), and 40, at a control temperature of 17 °C) or to two temperatures (17 and 22 °C, at the control salinity of 30). Under these conditions, Y bioaccumulation and different biomarkers were evaluated. Results showed that salinity and temperature did not affect Y accumulation, indicating effective detoxification mechanisms and physiological adaptations in the exposed organisms. However, in Y-exposed mussels effects were intensified under decreased salinity, evidenced by increased metabolism, defense enzyme activities, and acetylcholinesterase (AChE) levels. Similar responses occurred under heat stress with enhanced metabolic capacity, AChE activity, and activation of defense mechanisms such as glutathione S-transferases. These defense mechanisms mitigated cellular damage caused by Y, but under the highest temperature and especially lower salinity, Y-exposed mussels exhibited increased oxidative stress and decreased efficiency of activated defense enzymes, resulting in cellular damage compared to their uncontaminated counterpart. The present study sheds light on the effects that interactions between temperature, salinity, and the presence of emerging contaminants like REEs may have on marine organisms. Such assessments are crucial for developing effective strategies to mitigate the impacts of CC and protect the long-term health and resilience of marine ecosystems.

Threats of Pollutants Derived from Electronic Waste to Marine Bivalves: The Case of the Rare-Earth Element Yttrium.

The production of electrical and electronic equipment waste (e-waste) is increasing at an alarming rate worldwide. This may eventually lead to its accumulation in aquatic environments, mainly because of the presence of nonbiodegradable components. The rare-earth element yttrium (Y) is particularly relevant because it is present in a wide variety of electro-based equipment. Within this context, the present study investigated the biological consequences of anthropogenic Y exposure in Mytilus galloprovincialis. Mussels were exposed to Y (0, 5, 10, 20, 40 µg/L) for 28 days, and their bioaccumulation and biomarkers related to metabolism, oxidative stress defenses, cellular damage, and neurotoxicity were evaluated. The results revealed that tissue Y content increased at increasing exposure concentrations (though the bioconcentration factor decreased). At the lowest Y dosage (5 µg/L), mussels lowered their electron transport system (ETS) activity, consumed more energy reserves (glycogen), and activated superoxide dismutase activity, thus preventing cellular damage. At the highest Y dosage (40 µg/L), mussels reduced their biotransformation activities with no signs of cellular damage, which may be associated with the low toxicity of Y and the lower/maintenance of ETS activity. Although only minor effects were observed, the present findings raise an environmental concern for aquatic systems where anthropogenic Y concentrations are generally low but still may compromise organisms' biochemical performance. Particularly relevant are the alterations in energy metabolism and detoxification processes for their longer-term impacts on growth and reproduction but also as defense mechanisms against other stressors. Environ Toxicol Chem 2023;42:166-177. © 2022 The Authors. Environmental Toxicology and Chemistry published by Wiley Periodicals LLC on behalf of SETAC.

Gadolinium accumulation and its biochemical effects in Mytilus galloprovincialis under a scenario of global warming.

Electrical and electronic equipment reaching the end of its useful life is currently being disposed of at such an alarmingly high pace that raises environmental concerns. Together with other potentially dangerous compounds, electronic waste contains the rare-earth element gadolinium (Gd), which has already been reported in aquatic systems. Additionally, the vulnerability of aquatic species to this element may also be modified when climate change related factors, like increase in temperature, are taken into consideration. Thus, the present study aimed to evaluate the toxicity of Gd under a scenario of increased temperature in Mytilus galloprovincialis mussels. A multi-biomarker approach and Gd bioaccumulation were assessed in mussels exposed for 28 days to 0 and 10 µg/L of Gd at two temperatures (control - 17 °C; increased - 22 °C). Results confirmed that temperature had a strong influence on the bioaccumulation of Gd. Moreover, mussels exposed to Gd alone reduced their metabolism, possibly to prevent further accumulation, and despite catalase and glutathione S-transferases were activated, cellular damage seen as increased lipid peroxidation was not avoided. Under enhanced temperature, cellular damage in Gd-exposed mussels was even greater, as defense mechanisms were not activated, possibly due to heat stress. In fact, with increased temperature alone, organisms experienced a general metabolic depression, particularly evidenced in defense enzymes, similar to the results obtained under Gd-exposure. Overall, this study underlines the importance of conducting environmental risk assessment taking into consideration anticipated climate change scenarios and exposures to emerging contaminants at relevant environmental concentrations.

Assessing the impact of terbium on Mytilus galloprovincialis: Metabolic and oxidative stress responses.

The increasing demand for electric and electronic equipment has led to a rise in potentially hazardous electronic waste, including rare-earth elements (REEs), such as terbium (Tb), which have been already detected in aquatic systems. This study investigated the biochemical effects of anthropogenic Tb on mussels over a 28-day period. The mussels were exposed to different concentrations of Tb (0, 5, 10, 20, 40 µg/L), and biomarkers related to metabolism, oxidative stress, cellular damage, and neurotoxicity were evaluated. Bioaccumulation of Tb in the mussels' tissue increased with exposure concentrations, but the bioconcentration factor remained similar between treatments. Exposure to Tb enhanced glycogen consumption and decreased metabolic capacity which could be seen as a physiological adaptation to limit Tb accumulation. Antioxidant defenses and glutathione S-transferases showed a more complex dose-response, with enzymatic responses increasing until 10 µg/L but then returning to control levels at 20 µg/L. At 40 µg/L, enzymatic responses were also enhanced but to a lower extent than at 10 µg/L. The presence of Tb had clearly an inhibitory effect on biotransformation enzymes such as carboxylesterases in a dose-dependent manner. Likely, thanks to biochemical and physiological adaptations, no cellular damage or neurotoxicity was observed in any treatments, confirming the mussels' ability to tolerate Tb exposure. Nevertheless, prolonged exposure to these concentrations could lead to harmful consequences when facing other environmental stressors, such as misallocating energy resources for growth, reproduction, and defense mechanisms.

Do climate change related factors modify the response of Mytilus galloprovincialis to lanthanum? The case of temperature rise.

Most of the electric and electronic waste is not recycled and the release of its components into the environment is expected, including the rare-earth element Lanthanum (La), which has already been reported in the aquatic systems. Furthermore, considering climate change factors such as the predicted increase in temperature, the susceptibility of aquatic organisms to these rare elements may be modified. In light of this, the present study aimed to evaluate the relevance of temperature on La-derived effects in the mussel Mytilus galloprovincialis. Several biomarkers and La bioaccumulation were assessed in organisms exposed to 0 (control) and 10 µg/L of La at two distinct temperatures (17 and 22 °C) for 28 days. Results showed that temperature did not influence La bioaccumulation in mussels. However, exposure to La resulted in a decreased metabolic capacity and an enhancement of biotransformation enzymes activity, as a possible defense behavior of mussels to avoid La accumulation and toxicity. Nevertheless, antioxidant defenses were also inhibited leading to increased lipid peroxidation (LPO) levels. Warming alone seemed to cause a metabolic shutdown seen as reduced enzyme activities and protein carbonylation (PC) levels. Simultaneous La exposure and temperature rise caused combined effects on mussels, as they accused metabolic depression, biotransformation defenses activation, antioxidant capacity reduction, and higher cellular damage. Overall, this study highlights the need to perform environmental risk assessment studies, by considering emerging contaminants exposures at relevant concentrations, both at present and forecasted climate change scenarios.

Will climate changes enhance the impacts of e-waste in aquatic systems?

The increase of the worlds' population is being accompanied by the exponential growth in waste of electrical and electronic equipment (e-waste) generation as a result of the rapid technological implementations. The inappropriate processing and disposal of this e-waste, containing rare-earth elements (REEs) such as gadolinium (Gd), may enhance its occurrence in the environment. In particular, the presence of Gd in marine systems may lead to environmental risks which are still unknown, especially considering foreseen climate modifications such as water salinity shifts due to extreme weather events. Within this context, the present study intended to assess the combined effects of Gd at variable salinities. For that, biochemical modifications were assessed in mussels, Mytilus galloprovincialis, exposed to Gd (0 and 10 µg/L) and different salinity levels (20, 30 and 40), acting individually and in combination. A decrease in salinity, induced an array of biochemical effects associated to hypotonic stress in non-contaminated and contaminated mussels, including metabolism, antioxidant and biotransformation defenses activation. Moreover, in Gd-contaminated organisms, the increase in salinity was responsible for a significant reduction of metabolic and defense mechanisms, possibly associated with a mussels' physiological response to the stress caused by the combination of both factors. In particular, Gd caused cellular damage at all salinities, but mussels adopted different strategies under each salinity to limit the extent of oxidative stress. That is, an increase in metabolism was associated to hypotonic stress and Gd exposure, an activation of defense enzymes was revealed at the control salinity (30) and a decrease in metabolism and non-activation of defenses, associated with a possible physiological defense trait, was evidenced at the highest salinity. The different strategies adopted highlight the need to investigate the risk of emerging contaminants such as REEs at present and forecasted climate change scenarios, thus providing a more realistic environmental risk assessment.

Salinity influences on the response of Mytilus galloprovincialis to the rare-earth element lanthanum.

The multiplicity and wide variety of applications of electrical and electronic equipment has largely increased with the technological and economic progress and, in consequence, the amount of generated waste of electrical and electronic equipment (WEEE). Due to inappropriate processing and disposal of WEEE, different chemical elements and compounds, including rare-earth elements such as Lanthanum (La) have been released in the environment. Nevertheless, the environmental risks resulting from La presence are almost unknown, especially in marine systems, which may be challenged by foreseen climate changes such as water salinity shifts. Within this context, the present study aimed to understand the combined effects of salinity and La by assessing biochemical alterations in mussels Mytilus galloprovincialis exposed to La (0 and 10 µg/L) at different salinity levels (20, 30 and 40). A decrease in salinity caused a wide range of biochemical changes to both non-contaminated and contaminated organisms, such as metabolism, antioxidant and biotransformation defenses activation, associated to hypotonic stress. Furthermore, the decrease in salinity enhanced the effects of La exposure seen as an increase on lipid and protein cellular damage in those exposed, probably due to free metal ions increase at lower salinities, resulting in a higher bioaccumulation and toxicity. In general, La exposure caused cellular damage and inhibition of antioxidant defenses in contaminated mussels when compared to non-contaminated ones, with cellular damages being higher at the lowest salinity. Overall, the present study highlights the need to investigate the presence and impacts of emerging contaminants of WEEE source at environmental relevant concentrations, not just at present but also under forecasted climate change scenarios, thus providing a more realistic environmental risk assessment.

Are we neglecting earth while conquering space? Effects of aluminized solid rocket fuel combustion on the physiology of a tropical freshwater invertebrate.

Space launchers often use aluminized-solid fuel ("propergol") as propellant and its combustion releases tons of Al2O3 and HCl that sink in terrestrial and aquatic environments, polluting and decreasing water pH. We studied the impact of these events on the biochemical/physiological performance of the freshwater shrimp Macrobrachium jelskii, with wild specimens collected from a non-impacted site in French Guiana. In the laboratory, shrimps were exposed for one week to: i) undisturbed conditions; ii) Al2O3 exposure (0.5 mg L-1) at normal pH (6.6); iii) decreased pH (4.5) (mimicking HCl release in the environment) with no Al2O3; or iv) Al2O3 0.5 mg L-1 and pH 4.5, representing the average conditions found in the water bodies around the Ariane 5 launch pad. Results showed that shrimps bioaccumulated aluminium (Al) regardless of water pH. The combined effect of Al2O3 and low pH caused the most impact: acetylcholinesterase and carboxylesterase activities decreased, indicating neurotoxicity and reduced detoxification capacity, respectively. Animal respiration was enhanced with Al2O3 and pH variations alone, but the synergic interaction of both stressors caused respiration to decrease, suggesting metabolic depression. Oxidative damage followed a similar pattern to respiration rates across conditions, suggesting free radical-mediation in Al toxicity. Antioxidant activities varied among enzymes, with glutathione reductase being the most impacted by Al2O3 exposure. This study shows the importance of addressing space ports' impact on the environment, setting the bases for selecting the most appropriate biomarkers for future monitoring programs using a widespread and sensitive crustacean in the context of an increasing space-oriented activity across the world.

Does salinity variation increase synergistic effects of triclosan and carbon nanotubes on Mytilus galloprovincialis? Responses on adult tissues and sperms.

The use of carbon nanotubes (CNTs) is rapidly increasing and several scientific studies have addressed their toxicological properties. However, only a very small number of publications have deal with the interaction between CNTs and other molecules. Triclosan (TCS) is an antibacterial agent used in personal care and household products. Commonly detected in aquatic ecosystems, there is a strong evidence that aquatic biota is sensitive to this compound. Aside from emergent pollutants, aquatic organisms are continuously subjected to abiotic variations including salinities. Therefore, the main goal of the present study was to better understand how physio-chemical interactions of CNTs with TCS under different salinity levels (37, 28 and 19) affect the mussel species Mytilus galloprovincialis through the evaluation of biochemical alterations on gametes (sperms) and adult tissues, providing more ecologically relevant information on organisms' responses. The results showed toxicological effects in terms of sperm metabolic activity and intracellular reactive oxygen species production as well as cellular damage and alteration of metabolic capacity at the adult's stage when exposed to both contaminants acting alone and in combination, under tested salinities. Moreover, when the mussels were exposed to the combination of both contaminants, they showed major toxic impacts on both assessed biological levels (adult tissues and sperms) especially under control salinity. This suggests that toxicity upon mixture exposure compared to single-substance exposure may impair mussels' populations, affecting reproduction success and growth.

Are the effects induced by increased temperature enhanced in Mytilus galloprovincialis submitted to air exposure?

Intertidal mussel species are frequently exposed to changes of environmental parameters related to tidal regimes that include a multitude of stressors that they must avoid or tolerate by developing adaptive strategies. In particular, besides air exposure during low tides, intertidal mussels are also subjected to warming and, consequently, to higher risk of desiccation. However, scarce information is available regarding the responses of mussels to tidal regimes, particularly in the presence of other stressors such as increased temperature. Investigating the impacts of such combination of conditions will allow to understand the possible impacts that both factors interaction may generate to these intertidal organisms. To this end, the present study evaluated the impacts of different temperatures (18 °C and 21 °C) on Mytilus galloprovincialis when continuously submersed or exposed to a tidal regime for 14 days. Results showed that in mussels exposed to increased temperature under submersion conditions, the stress induced was enough to activate mussels' antioxidant defenses (namely glutathione peroxidase, GPx), preventing oxidative damage (lipid peroxidation, LPO; protein carbonylation, PC). In mussels exposed to tides at control temperature, metabolic capacity increased (electron transport system activity, ETS), and GPx was induced, despite resulting in increased LPO levels. Moreover, the combination of tides and temperature increase led to a significant decrease of lipid (LIP) content, activation of antioxidant defenses (superoxide dismutase, SOD; GPx) and increase of oxidized glutathione (GSSG), despite these mechanisms were not sufficient to prevent increased cellular damage. Therefore, the combination of increased temperature and air exposure induced higher oxidative stress in mussels. These findings indicate that increasing global warming could be more impacting to intertidal organisms compared to organisms continuously submersed. Furthermore, our results indicate that air exposure can act as a confounding factor when assessing the impacts of different stressors in organisms living in coastal systems.

The impacts of warming on the toxicity of carbon nanotubes in mussels.

With the increased production and research on nanoparticles, the presence of carbon nanotubes (CNTs) in aquatic systems is very likely to increase. Although it has been shown that CNTs may cause toxicity in marine organisms, to our knowledge, the possible impacts under global temperature increase is still unknown. For this reason, biochemical and physiological impacts induced in Mytilus galloprovincialis due to the presence of functionalized multi-walled CNTs (f-MWCNTs) and increased temperature were investigated in the present study. The mussels exposed to increased temperature alone presented higher metabolic capacity and expenditure of glycogen as an energy resource to fuel up defense mechanisms and thus preventing oxidative damage. Contrarily, organisms exposed to f-MWCNTs alone seemed not stressed enough to demonstrate differences in the metabolism capacity. Furthermore, f-MWCNTs seemed not able to significantly activate their antioxidant and biotransformation enzymes, which in turn may led to oxidative damage in the cells especially when organisms were exposed to a warmer temperature. In fact, at higher temperature, the antioxidant response of organisms exposed to f-MWCNTs was not effective and oxidative damage levels were observed. Nevertheless, no additive or synergetic effects were observed when mussels were exposed to both stressors simultaneously.

Biochemical changes in mussels submitted to different time periods of air exposure.

Intertidal species face multiple stressors on a daily basis due to their particular habitat. The submergence at high tide in the aquatic environment and emergence at low tide to the aerial environment, associated with a wide variation of abiotic parameters, along with anthropogenic contamination are some of the daily stresses that these organisms are exposed to. With such a dynamic environment, organisms developed strategies that allow them to avoid or tolerate these stressors. Among these species, bivalves are some of the most hypoxia tolerant, being commonly used as a biomonitoring tool due to their capacity to accumulate pollutants from the environment and reflect the imposed toxic impacts. However, when evaluating the response ability of organisms to different stressors under laboratory conditions, it is not common to consider the fact that exposure to tides can act as a confounding factor. The present study assessed the effects of air exposure on the biochemical (metabolic capacity, energy reserves, and oxidative stress related biomarkers) performance of intertidal Mytilus galloprovincialis mussels. Specimens of M. galloprovincialis were submitted once every 24 h to different periods of air exposure (3 and 6 h) for 14 days, under constant air and seawater temperature (19 ± 1 °C). Results obtained revealed that air exposure can cause biochemical changes in mussels. The present findings demonstrated that individuals exposed to air induced superoxide dismutase (SOD) and catalase (CAT) activity as mechanisms to withstand the abiotic changes while mobilizing lipid content as the principal source of energy, and increasing protein content possibly as a result of an increase in the number of antioxidant defense enzymes. Moreover, individuals under air exposure suffered higher oxidative damage while showing higher metabolic rate. Results demonstrated that longer periods of air exposure induced more injuries, since individuals emerged during 6 h presented higher oxidative stress than individuals under 3 h of air exposure.

Are the impacts of carbon nanotubes enhanced in Mytilus galloprovincialis submitted to air exposure?

Intertidal species are frequently exposed to environmental changes associated with multiple stressors, which they must either avoid or tolerate by developing physiological and biochemical strategies. Some of the natural environmental changes are related with the tidal cycle which forces organisms to tolerate the differences between an aquatic and an aerial environment. Furthermore, in these environments, organisms are also subjected to pollutants from anthropogenic sources. The present study evaluated the impacts in Mytilus galloprovincialis exposed to multi-walled carbon nanotubes (0.01 mg/L MWCNTs) when continuously submersed or exposed to tides (5 h of low tide, 7 h of high tide) for 14 days. Our results demonstrated that mussels were physiologically and biochemically affected by MWCNTs, especially when exposed to tides. In fact, when only exposed to the carbon nanoparticles or only exposed to tides, the stress induced was not enough to activate mussels' antioxidant defenses which resulted in oxidative damage. However, when mussels were exposed to the combination of tides and MWCNTs increased metabolism was observed, associated with a possible higher production of reactive oxygen species (ROS), leading to a significant increase in the activities of antioxidant enzymes (superoxide dismutase, SOD and glutathione peroxide, GPx) and oxidized glutathione content (GSSG), preventing the occurrence of cellular damage, expressed as no lipid peroxidation (LPO) or protein carbonylation (PC). Therefore, organisms seemed to be able to tolerate MWCNTs and air exposure during tidal regime; however, the combination of both stressors induced higher oxidative stress. These findings indicate that the increasing presence of carbon nanoparticles in marine ecosystems can induce higher toxic impacts in intertidal organisms compared to organisms continuously submerged. Also, our results may indicate that air exposure can act as a cofounding factor on the assessment of different stressors in organisms living in coastal systems.