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Does temperature influence on biomarker responses to copper exposure? The invasive bivalve Limnoperna fortunei (Dunker 1857) as a model
Abstract
Biomarkers are useful tools for assessing the early warning effects of pollutants. However, their responses can be influenced by confounding factors. In this study, we investigated the influence of temperature on multiple biomarkers in the invasive freshwater bivalve Limnoperna fortunei exposed to copper (Cu). The mussels were exposed to low and high environmental Cu concentrations at two temperatures (15°C and 25°C). After 96 h, the oxidative stress, neurotoxicity, and metabolic parameters were assessed. Our results showed that temperature is a key factor influencing biomarker responses in mussels, with higher glutathione S-transferase activity and lower energy reserves at cold temperature. In addition, the effects of Cu were greater at the highest concentration at 15°C (increased lipid peroxidation and cholinesterase activity). Overall, these findings suggest that cold stress increases the susceptibility of L. fortunei to metal effects and highlight the importance of including temperature in toxicity testing and biomonitoring. In addition, using the invasive bivalve L. fortunei as a model could prove valuable in its role as a sentinel species for other organisms.
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Invasive alien species (IAS) exert a negative impact on native ecosystems and on various human activities. Limnoperna fortunei (Dunker 1857), a sessile mytilid introduced from Asia in the Río de la Plata, demonstrates a high dispersal capacity, growing over other organisms and artificial structures. Understanding its behavior is crucial for developing appropriate control and mitigating its detrimental effects. This study investigated the population dynamics of this mollusk in response to urban effluents in a large river. Water and population parameters of L. fortunei were measured both upstream and downstream of a small town from July to December 2020. Downstream areas exhibited increased temperature and decreased transparency compared to upstream locations, while pH, conductivity, and dissolved oxygen remained relatively constant. However, downstream, the L. fortunei population comprised fewer and smaller individuals, displaying high width-to-length ratios. Most measured water parameters exhibited significant differences between sampling months. The abundance of L. fortunei remained relatively stable over time due to its nearly continuous reproduction in Uruguay. Nonetheless, shell elongation (15.3–22.8 mm) coincided with an increase in dry weight (0.009–0.038 g) from winter to spring. Allometric relationships, primarily following a power-law model, varied between months, ranging from H = 0.39L1.04 to H = 1.36L0.57. This study contributes to the comprehension of the population dynamics and life history of L. fortunei, thereby aiding in the conservation of invaded aquatic systems. Moreover, it provides valuable insights into the effects of urban effluents on this species, contributing to our understanding of its potential role as a bioindicator.
Surfactants are among the most common PPCPs that reach coastal systems, being often used in large quantities in cleaning products such as detergents and soap powders. Sodium lauryl sulfate (SLS) is listed in this group of emerging contaminants. Previous studies have already demonstrated the presence of SLS in aquatic environments and the negative effects on organisms living there. However, with ocean acidification and warming predictions, SLS-induced impacts may differ from those currently known. In this context, the present study aimed to reproduce environmental conditions by assessing the release of substances over a short period and to understand the influence of a rapid increase in temperature on the impacts caused. The marine bivalve Mytilus galloprovincialis was exposed to 2.0 mg/L SLS at 17 °C and 21 °C for 7 days. To assess the possible biochemical changes resulting from the exposure of mussels to SLS, a series of biomarkers related to oxidative stress/damage, detoxification, and metabolic capacity were measured. The SLS accumulation in soft tissues was low (about 0.7 ng/g) at both temperatures. The results evidenced increased metabolic activity, especially in mussels exposed to SLS at 17 °C. An increase in protein content was also observed upon exposure to SLS and increased temperature compared to controls at 17 °C. Although no effects on antioxidant enzymes were observed, protein damage was recorded, especially at 21 °C. These findings confirmed that SLS induces toxic effects and predicted climate change factors may increase the impact on M. galloprovincialis.
Biological early warning systems (BEWSs) monitor the behaviour or physiology of living organisms as an indirect mechanism to sense local environmental changes, and have become a widely established tool for monitoring water pollution. Complementary to conventional chemical and physical techniques, their strength lies in the ability to continuously monitor water quality, providing direct and rapid warning of toxic discharges. Bivalve molluscs (mussels and clams) are ideal sensing organisms for BEWSs, owing to their: high water filtration capacity; sensitivity to numerous, diverse pollutants; considerable longevity; high abundance; and wide range of physical responses to environmental change. Two behavioural metrics of bivalves have dominated in ecotoxicological studies, and consequently spawned development into BEWSs: valve movements, typically measured using electromagnetic techniques; and bivalve cardiac activity, typically measured using infrared photoplethysmography. However, other, less studied bivalve behaviours may exhibit pollutant impacts, including shell growth and dissolution, premature egg/larval release, horizontal movement, and burial. Such metrics may, with further research, provide bases for future BEWSs, some through current valve movement monitoring technology, others potentially through acoustic telemetry. Future bivalve-based BEWSs may become more informative and sensitive through the greater nuance that lies in integration of different metrics simultaneously and/or multi-species monitoring. Future directions may seek techniques that reduce disturbance to bivalves and enable observation of a more natural mode of behaviour, through using untethered, or entirely unequipped, specimens. With rigorous controls for individual- and population-level variation, bivalves have great power as biosensors and, bolstered by the scope for future advancements, this field has the potential to make significant contributions to future water quality management.
The pollution of coastal water areas by heavy metals is constantly growing; therefore, the study of the mechanisms of impact of these toxicants on the organisms of hydrobionts is a topical direction of toxicology. Particularly pertinent are questions about the state of the reparation system in the aquatic organisms, which make it possible to assess the resistance, survival of hydrobionts, and the probability of remote consequences under the impact of heavy metals. Therefore, in this work, we investigated genome integrity and DNA repair ability in the gill cells of Mytilus trossulus, under conditions of copper (Cu²⁺) accumulation, and in the process of tissue depuration from this metal. Although the biochemical detoxification system was functioning, it is noted that destructive processes developed, including the accumulation of lipid peroxidation products (MDA) and DNA damage. It was also found that in all of the experimental groups of Mytilus trossulus, in the process of depuration from Cu²⁺ for 24 h, the levels of DNA damage and MDA content were markedly reduced, and further reduction was less intense.
A comprehensive understanding of chemical toxicity and temperature interaction is essential to improve ecological risk assessment under climate change. However, there is only limited knowledge about the effect of temperature on the toxicity of chemicals. To fill this knowledge gap and to improve our mechanistic understanding of the influence of temperature, the current study explored toxicokinetics and the chronic toxicity effects of two insecticides, imidacloprid (IMI) and flupyradifurone (FPF), on Gammarus pulex at different temperatures (7–24 °C). In the toxicokinetics tests, organisms were exposed to IMI or FPF for 2 days and then transferred to clean water for 3 days of elimination at 7, 18, or 24 °C. In the chronic tests, organisms were exposed to the individual insecticides for 28 days at 7, 11, or 15 °C. Our research found that temperature impacted the toxicokinetics and the chronic toxicity of both IMI and FPF, while the extent of such impact differed for each insecticide. For IMI, the uptake rate and biotransformation rate increased with temperature, and mortality and food consumption inhibition was enhanced by temperature. While for FPF, the elimination rate increased with temperature at a higher rate than the increasing uptake rate, resulting in a smaller pronounced effect of temperature on mortality compared to IMI. In addition, the adverse effects of the insecticides on sublethal endpoints (food consumption and dry weight) were exacerbated by elevated temperatures. Our results highlight the importance of including temperature in the ecological risk assessment of insecticides in light of global climate change.
Climatic change (global warming) not only limited to the terrestrial environmental variations but also transforming the aquatic environments. Recent studies have reported that both coastal and freshwater bodies have experienced the progressive warming and will intensify (giant leap) during this century. Same time the bewildering array of thermal stresses poses serious threats to the aquatic organisms. Although, there is a growing body of literature and scientific consensus regarding the effects of temperature to the aquatic biota. Hitherto, the extent to which the thermo-chemical stress on aquatic organisms intensifying is not fully elaborated. We have summarized following declarations:
• Temperature variations induce a range of physiological and biological responses in aquatic organisms including corticosteroid response, metabolic response, immune responses, heat shock proteins, and wavering in haemolymph parameters.
• The observed responses due to temperature fluctuations are actually attributed to the whole organism level performances like, acclimation and adaptation, fitness, feeding and food conversion ratio, growth and development.
• Temperature dependent chemical toxicities are subjected to influence the chemical processes and underlying molecular/genetic mechanisms of aquatic organisms at individual population and community levels.
Additionally, this review outlined the series of mechanisms in aquatic organisms associated with temperature changes in single or combined with chemicals. These observations provide significant evidence that the climate change and temperature variations are critical and there is pressing need to carefully evaluate the conditions and responses at large geographical scales.
Studies of changes in fatty acids in response to environmental temperature changes have been conducted in many species, particularly mammals. However, few studies have considered aquatic invertebrates, even though they are particularly vulnerable to changes in environmental temperature. In this review, we summarize the process by which animals synthesize common fatty acids and point out differences between the fatty acid profiles of vertebrates and those of aquatic invertebrates. Unlike vertebrates, some aquatic invertebrates can directly synthesize polyunsaturated fatty acids (PUFAs), which can be used to respond to temperature changes. Various studies have shown that aquatic invertebrates increase the degree of saturation in their fatty acids through an increase in saturated fatty acid production or a decrease in PUFAs as the temperature increases. In addition, we summarize recent studies that have examined the complex effects of temperature and combinational stressors to determine whether the degree of saturation in aquatic invertebrates is influenced by other factors. The combined effects of carbon dioxide partial pressure, food quality, starvation, salinity, and chemical exposures have been confirmed, and fatty acid profile changes in response to high temperature were greater than those from combinational stressors.
The purification of homogeneous glutathione S-transferases B and C from rat liver is described. Kinetic and physical properties of these enzymes are compared with those of homogeneous transferases A and E. The letter designations for the transferases are based on the reverse order of elution from carboxymethylcellulose, the purification step in which the transferases are separated from each other. Transferase B was purified on the basis of its ability to conjugate iodomethane with glutathione, whereas transferase C was purified on the basis of conjugation with 1,2-dichloro-4-nitrobenzene. Although each of the four enzymes can be identified by its reactivity with specific substrates, all of the enzymes are active to differing degrees in the conjugation of glutathione with p-nitrobenzyl chloride. Assay conditions for a variety of substrates are included.
All four glutathione transferases have a molecular weight of 45,000 and are dissociable into subunits of approximately 25,000 daltons. Despite the similar physical properties and overlapping substrate specificities of these enzymes, only transferases A and C are immunologically related.
The literature indicates that exotic species have a greater tolerance to environmental stressors compared with native species. In recent decades, the introduction of contaminants into the environment has increased as a result of industrialization. The objective of this study was to verify the resistance of bivalve mollusks from freshwater native (Anodontites trapesialis) and exotic (Limnoperna fortunei) species to chemical contamination using an ex vivo/in vitro approach. Gill and muscle tissues were exposed to two different types of environmental stressors, copper (metal), and Roundup Transorb® (herbicide). The tissues were submitted to a cytotoxicity test in which the lysosomal integrity was assessed, from the adaptation of a method to isolated cells, and multixenobiotic resistance (MXR) test which evaluated cellular defense. In the exotic species, only copper at 9000 μg/L and Roundup Transorb® at 5000 μg/L were cytotoxic. In the native species, copper cytotoxicity at 900 and 9000 μg/L and Roundup Transorb® at 50 and 5000 μg/L were observed. Results were the same in both tissues. The MXR, responsible for the extrusion of contaminants (cell defense), was inhibited in both species when exposed to the contaminants, this cell defense system seems to be more inhibited in the native species, when exposed to both pollutants, indicating greater sensitivity. Therefore, cytotoxicity may be related to the lack of capacity of cellular defense. In relation to lysosomal integrity, the native species was more sensitive to cytotoxic pollutants, where a greater number of experimental conditions of metals and herbicide showed cytotoxicity, as well as more experimental situations inhibited its ability to defend itself.
Heavy metals are the most dangerous hazards affecting aquatic biota in Egypt specially the Nile Tilapia, Oreochromis niloticus, which is an important species in commercial fisheries. Some areas were not fully studied to screen the hazards that may affect this economic fish. Therefore, the present study aimed to evaluate the potential hazards of heavy metals on O. niloticus in Al-Gharbiya Governorate in the Middle delta of Egypt. Water and fish samples were collected from Al-Qased canal, Kafr El-Zayaat Nile, El-Maash canal in Al-Gharbiya Governorate plus a reference site which is a fish farm at the Faculty of Agriculture, Damietta University, Damitta Governorate, Egypt. The results showed a significant increase (p < 0.05) in the lead, zinc, magnesium, manganese, and copper levels while showed a significant decrease (p < 0.05) in the iron level in the water from all the investigated areas. Cadmium level was significantly high (p < 0.05) in Kafr El-Zayaat Nile and EL-Maash canal only. These metals were also accumulated in the fish livers and gills. Consequently, ALT and AST activities and creatinine level were significantly (p < 0.05) high in all the investigated areas. Histopathological examination revealed cytoplasmic and nuclear degeneration in the hepatocytes in all the investigated areas. Renal glomeruli and Bowman’s capsule were not completely intact in Al-Qased and El-Maash canals while conspicuous shrinkage of the glomeruli was determined in Kafr El-Zayaat Nile. Furthermore, slight damage in the secondary lamellae was detected in the gill from Al-Qased canal while in the other areas appeared vacuolated or destructed. Finally, spleen sections of fish from different sites showed the absence of melanin pigments and some vacuoles. In conclusion, the Nile Tilapia, Oreochromis niloticus, is affected by the toxic effects of the heavy metals in Al-Gharbiya Governorate in Egypt and this gives an alarm and should be taken into consideration.
Limnoperna fortunei (Dunker 1857) is a freshwater mussel with physiological tolerance to different environmental conditions, which may explain its success as an invasive species. The role of abiotic factors in its establishment, abundance and projections of risk of further spread into several areas has been studied. These mussels may respond to multiple environmental stressors, such as temperature, through physiological mechanisms, behavioral responses, mortality or some combination of these. The aim of this study was to investigate the behavioral responses (valve closing), glycogen concentrations and mortality of L. fortunei under four different temperatures (5°C, 10°C, 20°C and 30° C) during a chronic test (30 days). Two-way analysis of variance (ANOVA) was used to compare glycogen concentrations across days of the experiment and at the different temperatures. Differences in valve-closing behavior and mortality among temperatures were tested using repeated-measures ANOVA. We observed that most of the mussels maintained at 5°C closed their valves (74.7 ± 15.3%), indicating that they remain inactive at low temperatures. The glycogen levels significantly differed among the temperatures tested. These differences occurred mainly due to the high glycogen values observed in mussels exposed to 10°C. Stability in glycogen concentrations was observed within each particular temperature. The cumulative mortality was higher at extreme temperatures (5°C and 30°C). The ideal temperature for laboratory maintenance and tests is approximately 20°C. Our data also show that L. fortunei can survive and maintain their energy reserves (glycogen) for several days at 5°C, an important feature related to its invasion success.
The present study focuses on the evaluation of metal (chromium, copper, and lead), arsenic, and pesticide (atrazine and endosulfan) contamination in freshwater streams of one of the most important agricultural and industrial areas of central-eastern Argentina, which has not been reported earlier. The environmental fate of inorganic microcontaminants and pesticides was assessed. Samples were collected monthly for a year. Pesticide concentrations were measured in water; metal and arsenic concentrations were measured in water and sediments, and physicochemical variables were analyzed. In most cases, metals and arsenic in water exceeded the established guideline levels for the protection of aquatic biota: 98 and 56.25% of the samples showed higher levels of Cr and Pb, while 81.25 and 85% of the samples presented higher values for Cu and As, respectively. Cr, Pb, Cu, and As exceeded 181.5 times, 41.6 times, 57.5 times, and 12.9 times, respectively, the guideline level values. In sediment samples, permitted levels were also surpassed by 40% for Pb, 15% for As, 4% for Cu, and 2% for Cr. Geoaccumulation Index (Igeo) demonstrated that most of the sediment samples were highly polluted by Cr and Cu and very seriously polluted by Pb, which indicates progressive deterioration of the sediment quality. Atrazine never exceeded them, but 27% of the 48 water samples contained total endosulfan that surpassed the guidelines. The findings of this study suggest risk to the freshwater biota over prolong periods and possible risk to humans if such type of contaminated water is employed for recreation or human use. Improper disposal of industrial effluents and agricultural runoffs need to be controlled, and proper treatment should be done before disposal to avoid further deterioration of the aquifers of this area.
Pre-spawning blue mussels (Mytilus edulis) appear sensitive to elevated temperature and robust to elevated pCO2; however, the effects of these stressors soon after investing energy into spawning remain unknown. Furthermore, while studies suggest that elevated pCO2 affects the byssal attachment strength of Mytilus trossulus from southern latitudes, pCO2 and temperature impacts on the byssus strength of other species at higher latitudes remain undocumented. In a 90 day laboratory experiment, we exposed post-spawning adult blue mussels (Mytilus edulis) from Atlantic Canada to three pCO2 levels (pCO2 ~625, 1295, 2440 μatm) at two different temperatures (16 ºC and 22 ºC) and assessed energetic reserves on day 90, byssal attachment strength on days 30 and 60, and condition index and mortality on days 30, 60 and 90. Results indicated that glycogen content was negatively affected under elevated temperature, but protein, lipid, and overall energy content were unaffected. Reductions in glycogen content under elevated temperature was associated with reduced condition index, reduced byssal thread attachment strength, and increased mortality; elevated pCO2 had no effects. Overall, these results suggest that the glycogen reserves of post-spawning adult M. edulis are sensitive to elevated temperature, and can result in reduced health and byssal strength, leading to increased mortality. These results are similar to those reported for pre-spawning mussels and suggest that post-spawning blue mussels are tolerant to elevated pCO2 and sensitive to elevated temperature. In contrast to previous studies, however, elevated pCO2 did not affect byssus strength, suggesting that negative effects of elevated pCO2 on byssus strength are not universal.
Most molluscs have blue blood because their respiratory molecule is hemocyanin, a type-3 copper-binding protein that turns blue upon oxygen binding. Molluscan hemocyanins are huge cylindrical multimeric glycoproteins that are found freely dissolved in the hemolymph. With molecular masses ranging from 3.3 to 13.5 MDa, molluscan hemocyanins are among the largest known proteins. They form decamers or multi-decamers of 330- to 550-kDa subunits comprising more than seven paralogous functional units. Based on the organization of functional domains, they assemble to form decamers, di-decamers, and tri-decamers. Their structure has been investigated using a combination of single particle electron cryo-microsopy of the entire structure and high-resolution X-ray crystallography of the functional unit, although, the one exception is squid hemocyanin for which a crystal structure analysis of the entire molecule has been carried out. In this review, we explain the molecular characteristics of molluscan hemocyanin mainly from the structural viewpoint, in which the structure of the functional unit, architecture of the huge cylindrical multimer, relationship between the composition of the functional unit and entire tertiary structure, and possible functions of the carbohydrates are introduced. We also discuss the evolutionary implications and physiological significance of molluscan hemocyanin.
In the context of environmental monitoring studies bioindicators reflect organisms (or parts of organisms or communities of organisms) that contain information on quality of the environment (or a part of the environment). Biomonitors, on the other hand, are organisms (or parts
of organisms or communities of organisms) that contain informations on the quantitative aspects of quality of the environment. When data and information obtained by bioindication are moved up to the level of knowledge the subjectivity of interpretation increases with the
complexity and dynamics of a system (“staircase of knowing”).
In this article clearcut definitions are attempted for most terms used in environmental monitoring studies. From there a comparison of instrumental measurements with the use of bioindicators/biomonitors with respect to harmonisation and quality control will be drawn.
Precision, accuracy, calibration and harmonisation in between national standards and international routines seem to be the leading goals in quality studies of international working groups dealing with biomonitoring throughout the world. Common strategies and concepts
will fill the gap in between single source results and integrated approaches related either for human health aspects or environmental protection purposes, f.e. via biodiversity monitoring. Here we report on well established monitoring programmes like Environmental Specimen
Banking (ESB) or newly developed strategies as the Multi-Markered Bioindication Concept (MMBC) with its functional and integrated windows of prophylactic health care.
Full-text access: https://authors.elsevier.com/a/1hqpHB8ccyf4B Pollution of surface waters is a global threat, with particular concern about pesticides due to their severe negative effects on ecosystem functioning and human health. The aims of this study were to identify the spatiotemporal patterns of water and sediment quality, and the key variables related to the variation in pesticide pollution, in headwater streams (surrounding land uses: crop or mixed crop-livestock systems) and floodplain streams (surrounding land uses: urban development or natural wetland) of the Paraná River basin in the central area of Argentina. We found significant differences in water and sediment quality related to local land uses among headwater streams, but not among floodplain streams. These differences were more noticeable during spring than during autumn. Pesticides were widespread in all the streams, independently of the surrounding land use, reflecting the combination of local inputs and the role of floodplain hydrological connectivity in transporting pollutants from upstream sources. The most frequently detected compound was atrazine (75 %), whereas the highest concentration of an individual compound was observed for the glyphosate metabolite aminomethylphosphonic acid (AMPA, up to 4 μg L−1). The significant explanatory variables for pesticide pollution were turbidity, chromophoric dissolved organic matter (CDOM), sub-basin area, side slope of streams (positive relations), wetland cover, and precipitations (negative relations). Our results can be useful for the design of monitoring programs that capture the spatial and temporal variability of pesticide pollution.
Lead and Lithium are metals which have been detected in the environment and, at high concentrations, can induce toxic effects that disturb the growth, metabolism or reproduction of organisms along the entire trophic chain. The impacts of these metals have scarcely been investigated using marine bivalves, especially when acting as a mixture. The present study aimed to investigate the influence of temperature on the ecotoxicological effects caused by Pb and Li, acting alone and as a mixture, on the mussel species Mytilus galloprovincialis after 28 days of exposure. The impacts were evaluated under actual (17 °C) and projected (+4 °C) warming conditions, to understand the influence of temperature rise on the effects of the metals (both acting alone or as a mixture). The results obtained showed that the increased temperature did not influence the accumulation of metals. However, the biomarkers evaluated showed greater responses in mussels that are exposed to metals under increased temperature (21 °C). The IBR index showed that there is a comparable toxic effect of Li and Pb separately, while exposure to a mixture of both pollutants causes a significantly higher stress response. Overall, the results obtained revealed that temperature may cause extra stress on the mussels and exposure to the metal mixture caused the greatest impacts compared to each metal acting alone.
Intertidal mussels are well adapted to withstand emersion from water during low tide, but they may be intermittently exposed to waterborne toxicants such as copper, which targets physiological processes including metabolism, ammonia excretion, and osmoregulation. To determine if copper exposure damages intertidal organisms' ability to tolerate tidal emersion, Mediterranean mussels (Mytilus galloprovincialis) were exposed to copper for 96 h followed by 6 h of emersion. Oxygen uptake increased after copper exposure which suggests that copper accumulation caused moderate stress in the mussels, but ammonia excretion and anaerobic metabolism were unaffected by mixed copper and emersion exposures. Shell composition analyses indicate that cycles of copper exposure and tidal emersion may affect bivalve shell growth, but copper deposition into shells may decrease the metal's overall toxicity. Results suggest that copper does not damage M. galloprovincialis's tolerance to tidal emersion, and insight is provided into the mussel's ability to overcome mixed stressor exposures.
Cobalt (Co) is among the hazardous substances identified in aquatic environments. Industrialization and population growth have also contributed to climate change, namely in what concerns ocean temperature rise. The aim of the present study was to evaluate the influence of temperature rise on the impacts caused by Co on Mytilus galloprovincialis. To this end, mussels were exposed for 28 days to 17 °C and 21 °C, without and with 200 μg L⁻¹ of Co. Results showed no significant differences in Co bioaccumulation by the organisms between temperatures. A significant interaction between temperature and Co contamination was observed in terms of oxidative damage, detoxification capacity, and neurotoxicity, with a synergistic effect particularly evident in terms of biotransformation enzymes' activity. The obtained results point out that population survival and distribution may be limited in the long term, highlighting the need for future research on the combined effects of both stressors.
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 28days. 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.
Copper has become one of the most important heavy metal pollutants in the environment because of its wide application and high toxicity, but research on water quality criteria (WQCs) on copper is limited, especially the derivation of seawater WQC. In addition, the toxicity of copper in the seawater system is affected by various environmental factors. Therefore, establishing a WQC that meets the characteristics of the regional environment is a top priority. The correlations between four factors of temperature, salinity, pH, dissolved organic carbon (DOC) and the toxic effect values of copper were analyzed in this study, and the temperature was determined as the most influential factor among the four factors in the Bohai Sea. A specific correlation between temperature and the toxic effects of copper was identified, and WQCs were derived based on the identified correlation and the variations of the Bohai Sea's temperature in different seasons by species sensitivity distribution (SSD) method. Under the condition of the winter, spring, autumn, and summer with an average water temperature of 0.09, 15.96, 17.83, and 24.87 °C, the obtained short-term water quality criteria (SWQCs) were 44.29, 4.70, 4.31, and 3.33 μg/L; the long-term water quality criteria (LWQCs) were 18.14, 1.93, 1.77 and 1.36 μg/L. The findings indicated the importance of introducing specific environmental conditions during the derivation process. This work could provide valuable information for pollution prevention and aquatic life protection in the Bohai Sea and provide a valuable reference for the derivation of criteria in other regions alike.
Behavioral parameters are increasingly considered sensitive and early bioindicators of toxicity in aquatic organisms. A video-tracking tool was specifically developed to monitor the swimming behaviour of D-larvae of the Mediterranean mussel, Mytilus galloprovincialis, in controlled laboratory conditions. Both maximum and average swimming speeds and trajectories were recorded. We then investigated the impact of copper and silver with or without a moderate rise of temperature on swimming behavior and acetylcholinesterase (AChE) activity of mussel D-larvae and the possible mechanistic link between both biological responses. Our results showed that copper and/or silver exposure, as well as temperature increase, disrupts the swimming behavior of mussel larvae which could compromise their dispersal and survival. In addition, the combined effect of temperature and metals significantly (p < 0.05) increased AChE activity in mussel larvae. Pearson's correlation analysis was performed and results showed that the AChE activity is positively correlated with maximum speeds (r = 0.71, p < 0.01). This study demonstrates the value of behavioral analyzes of aquatic invertebrates as a sensitive and integrate marker of the effects of stressors.
Plasticity and variability of morphological and physiological traits are common characteristics that allow invasive species to survive, establish and spread to new environments. In order to study the metabolic response of the invasive mussel Limnoperna fortunei (Dunker) (Mytilidae), which has spread through South America and Asia, oxygen consumption was measured under three different acclimation temperatures (18, 22 and 27°C). After an acclimation period of at least 7 days, oxygen consumption was measured and analysed as functions of length and mass. Biochemical composition was also investigated on specimens collected under natural conditions. Environmental scanning electron microscopy determined that the elemental composition and crystalline structure of the shells were normal as described for L. fortunei previously. Average oxygen consumption per mussel was 1.63 ± 0.12 μmol O2 h−1 for an average ash‐free dry weight (AFDW) of 58.7 ± 3.0 mg. The average standard metabolic rate (SMR) per mussel was 14.94 ± 1.30, 30.04 ± 3.38, and 47.10 ± 3.54 μmol O2 gAFDW−1 h−1, at 18, 22 and 27°C, respectively. Significant higher SMR values were recorded at 27°C than at lower temperatures. This metabolic response allows L. fortunei to survive, reproduce and spread across a wide range of habitats, including tropical and subtropical environments in South America.
Acetylcholinesterase (AChE; EC 3.1.1.7) from aquatic organisms have been used to evaluate the exposure of specimens to pesticides and heavy metals at sublethal levels in environmental samples. AChE of Mytella charruana was extracted to characterize its physicochemical and kinetic properties as well as the effect of organophosphate (dichlorvos, diazinon, chlorpyrifos, methyl-parathion and temephos), carbamates (carbaryl, carbofuran and aldicarb), benzoylureas (diflubenzuron and novaluron), pyrethroid (cypermethrin) and juvenile hormone analog - JHA (pyriproxyfen) and the effect of metal ions: Hg²⁺, Cd²⁺, Pb²⁺, As³⁺, Cu²⁺ and Zn²⁺, in order to evaluate the potential of the enzyme as biomarker. The optimum pH of M. charruana AChE was 8.5 and the maximum activity peak occurred at 48 °C, being highly thermostable maintaining 97.8% of its activity after incubation at 60 °C. The Michaelis-Menten constants (km) for the substrates acetylthiocholine and propionylthiocholine were 2.8 ± 1.26 and 4.94 ± 6.9 mmol·L⁻¹, respectively. The Vmax values for the same substrates were 22.6 ± 0.90 and 10.2 ± 4.94 mU·mg⁻¹, respectively. Specific inhibition results suggest an AChE presenting active site with dimensions between those of AChE and butyrylcholinesterase (BChE). The IC20 values related to the effect of the pesticides on the enzyme showed higher inhibitory power of temephos (0.17 μmol·L⁻¹), followed by aldicarb (0.19 μmol·L⁻¹) and diflubenzuron (0.23 μmol·L⁻¹). Metal ions inhibited M. charruana enzyme in the following order: Hg²⁺ > Pb²⁺ > Cd²⁺ > As³⁺ > Cu²⁺ > Zn²⁺. These data suggest that the enzyme showed potential as in vitro biomarker of the exposure to temephos, mercury, zinc and copper.
Pharmaceuticals drugs are Contaminants of Emerging Concern (CECs) and are continuously discharged into the environment. As a result of human and veterinary use, these substances are reaching aquatic coastal systems, with limited information regarding the toxic effects of these compounds towards inhabiting organisms. Among CECs are pharmaceuticals like 17 alpha-ethinylestradiol (EE2), which is a synthetic hormone with high estrogenic potency. EE2 has been increasingly found in different aquatic systems but few studies addressed its potential toxicity to marine wildlife, in particular to bivalves. Therefore, the aim of the present study was to evaluate the influence of temperature (17 ºC-control and 21 °C) on the potential effects of EE2 on the mussel Mytilus galloprovincialis. For this purpose, mussels were exposed to different concentrations of EE2 (5.0; 25.0; 125.0 and 625 ng/L), resembling low to highly polluted sites. Mussels exposed to each concentration were maintained under two temperatures, 17 and 21 °C, which represent actual and predicted warming conditions, respectively. After 28 days, oxidative stress status, metabolism related parameters, neurotoxicity and histopathological alterations were measured. The results obtained clearly showed an interactive effect of increased temperature and EE2, with limited antioxidant and biotransformation capacity when both stressors were acting together, leading to higher cellular damage. The combination of both stressors also enhanced mussels’ metabolic capacity and neurotoxic effects. Nevertheless, loss of redox balance was confirmed by the strong decrease of the ratio between reduce glutathione (GSH) and oxidized glutathione (GSSG) in contaminated mussels, regardless the temperature. Histopathological indexes in contaminated mussels were significantly different from the control group, indicating impacts in gills and digestive glands of mussels due to EE2, with higher values observed at 21 °C. Overall, this study demonstrates that of EE2 represents a threat to mussels and predicted warming conditions will enhance the impacts, which in a near future might result in impairments at the population and community levels.
Contamination and biological invasions are important factors that alter the functioning of freshwater systems. We carried out two experiments involving daily measurements of dissolved oxygen (DO) and Limnoperna fortunei mortality: 1) the impact of DO on mussel survival mediated by glyphosate, 2,4-D and their mixture was analysed in a 22-day indoor experiment (IE) under controlled conditions using microcosms with L. fortunei, with and without air supply; and 2) the effect of each herbicide and their mixture on mussel accumulated mortality was compared in a 18-day outdoor experiment (OE) using mesocosms without air supply, with and without L. fortunei. Results showed that glyphosate, alone or mixed affected L. fortune survival both directly and indirectly. In IE we observed direct toxicity of glyphosate in treatments with air supply, with accumulated mortality of 20.0% for glyphosate and 10.0% for the mixture. In OE, L. fortunei deepened the changes in the patterns of DO fluctuations driven by the herbicides, which led to hypoxia in the system. The accumulated mortality was 46.7, 8.6 and 48.2% for glyphosate, 2,4-D and the mixture, respectively. This study contributes to the understanding of the mechanisms that control the invasion of L. fortunei in freshwater systems influenced by agrochemicals.
The Amazon is Earth's largest river by volume output, making it an important source of trace metals and dissolved organic matter (DOM) to the Atlantic Ocean. Despite major recent anthropogenic disruptions to the Amazon catchment area, data for trace metals such as copper (Cu) in the Amazon River estuary and associated mixing plume are still rare. Furthermore, there is currently no existing data in this region for Cu-binding ligands, which govern the amount of bioavailable Cu. To understand trace metal mixing and transport processes, the GEOTRACES process study GApr11 (cruise M147 with RV Meteor) was conducted in 2018 in the Amazon and Pará River estuaries and mixing plume in the tropical North Atlantic Ocean during high river discharge. Size-fractionated surface samples were collected along the full salinity gradient for concentrations of Cu, apparent Cu-binding organic ligands (LCu) and corresponding conditional stability constants (K′CuL, Cu2+cond), electroactive humic substances (eHS), solid phase extractable organic Cu (SPECu), dissolved organic carbon (DOC), chlorophyll a (Chl a) and macronutrients. Dissolved (<0.2 μm) and soluble (<0.015 μm) Cu correlated negatively with salinity and largely followed values expected from conservative mixing. Cu was primarily in the soluble fraction, with the exception of a minor fraction of large colloidal Cu at low salinity (S ≤ 10). Organic ligands (log K′CuL, Cu2+cond = 12.6–15.6) were present in excess of Cu and likely played a role in solubilizing Cu and preventing Cu being affected by colloidal flocculation. Cu-associated DOM (measured as LCu, eHS and SPECu) correlated negatively with salinity and appeared to be primarily governed by river input and mixing with seawater. However, an increase in the colloidal fraction for LCu and eHS observed at S ~ 6–10 was attributed to possible additional autochthonous (phytoplankton) ligand production. In all dissolved samples, organic complexation kept free Cu below levels potentially toxic for phytoplankton (<1 pmol L⁻¹). Despite increasing anthropogenic activity over the past century, we find Cu concentrations remained similar to the 1970s, suggesting that the large overall river flow may so far minimize the impact of Cu pollution.
We built a simulation model based on Dynamic Energy Budget theory (DEB) to assess the growth and reproductive potential of the native European clam Ruditapes decussatus and the introduced Manila clam Ruditapes philippinarum under current temperature and pH conditions in a Portuguese estuary and under those forecasted for the end of the 21st c. The climate change scenario RCP8.5 predicts temperature increase of 3°C and a pH decrease of 0.4 units. The model was run under additional conditions of exposure to the emerging contaminant triclosan (TCS) and in the absence of this compound. The parameters of the DEB model were calibrated with the results of laboratory experiments complemented with data from the literature available for these two important commercial shellfish resources. For each species and experimental condition (eight combinations), we used data from the experiments to produce estimates for the key parameters controlling food intake flux, assimilation flux, somatic maintenance flux and energy at the initial simulation time. The results showed that the growth and reproductive potential of both species would be compromised under future climate conditions, but the effect of TCS exposure had a higher impact on the energy budget than forecasted temperature and pH variations. The egg production of R. philippinarum was projected to suffer a more marked reduction with exposure to TCS, regardless of the climatic factor, while the native R. decussatus appeared more resilient to environmental causes of stress. The results suggest a likely decrease in the rates of expansion of the introduced R. philippinarum in European waters, and negative effects on fisheries and aquaculture production of exposure to emerging contaminants (e.g., TCS) and climate change.
This study aimed to estimate the impact of an abandoned copper (Cu) mine on ecosystem environmental quality, using integrated ecological and biogeochemical analyses. Through a controlled experiment, the amount of Cu released by waste rocks, Cu adsorbed in soils, Cu geochemical behaviour and its leached amount were measured. Furthermore, to investigate the impacts of mine drainage on the adjacent ecosystem, samples of sediments, water and aquatic macroinvertebrates were analysed. We found that waste rocks still have high Cu concentration even after 30 years under weathering, ranging from 7,782 to 8,717 mg kg⁻¹, associated mainly with carbonates, amorphous oxides and sulphides. It was estimated that 7.2 tonnes of Cu were released by waste rocks into the environment over last 30 years. The concentration of Cu observed in Ubari stream water was (<dl to 90 μg L⁻¹), in sediments (28.0 to 1,185 mg kg⁻¹) and in macroinvertebrates (1.3 to 28.9 mg kg⁻¹ d/w). The ecological indexes showed that near mine discharge a significance decrease in the density of aquatic macroinvertebrates and a significance increase of Cu in biological tissues occurs, causing disturbances in biodiversity. The results showed that, even after long periods, the waste rocks from abandoned mines still contain high levels of metal, that are gradually released into the environment through weathering and erosion, representing a potential source of environmental pollution and a clear threat to the environmental quality of adjacent ecosystems.
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 twenty-eight days 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 CeO2 NPs 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 decrease 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.
We determined values of antioxidant biomarkers catalase (CAT), superoxide dismutase (SOD), glutathione peroxidase (GPx), glutathione S-transferase (GST) and malondialdehyde (MDA) in two bivalve species from a subtropical system in Brazil. Differences in the biomarkers activities measured in digestive glands and gills sampled at two seasons and in sites with different levels of contamination were tested using univariate and multivariate analyses. Seasonality was the main factor accounting for overall variability. Significant SOD and GPx activities and increased levels of lipid peroxides were detected in summer, while CAT activity was higher in winter. However, biomarkers respond to complex environmental settings as indicated by the significant interaction between season and contamination level. We propose A. flexuosa as a candidate for monitoring studies with sampling occurring at a yearly timescale, and using C. rhizophorae is better suited for designs that include both seasons.
Environmental stressors induce changes in marine mussels from molecular (e.g., neurotransmitter and chaperone concentration, and expression of immune- and heat-shock protein-related genes) to physiological (e.g., filtration and heart rates, the number of circulating hemocytes) levels. Temperature directly affects the biogeographic distribution of mussels. Chaperones might form an essential part of endogenous protective mechanisms for the adaptation of these animals to low temperatures in nature. Here, we review the available studies dealing with cold stress responses of Mytilidae family members in their natural environment.
Ocean acidification and warming are among the man-induced factors that most likely impact aquatic wildlife worldwide. Besides effects caused by temperature rise and lowered pH conditions, chemicals of current use can also adversely affect aquatic organisms. Both climate change and emerging pollutants, including toxic impacts in marine invertebrates, have been investigated in recent years. However, less information is available on the combined effects of these physical and chemical stressors that, in nature, occur simultaneously. Thus, this study contrasts the effects caused by the antimicrobial agent and plastic additive, Triclosan (TCS) in the related clams Ruditapes philippinarum (invasive) and Ruditapes decussatus (native) and evaluates if the impacts are influenced by combined temperature and pH modifications. Organisms were acclimated for 30 days at two conditions (control: 17 °C; pH 8.1 and climate change scenario: 20 °C, pH 7.7) in the absence of the drug (experimental period I) followed by a 7 days exposure under the same water physical parameters but either in absence (unexposed) or presence of TCS at 1 μg/L (experimental period II). Biochemical responses covering metabolic, oxidative defences and damage-related biomarkers were contrasted in clams at the end of experimental period II. The overall picture showed a well-marked antioxidant activation and higher TCS bioaccumulation of the drug under the forecasted climate scenario despite a reduction on respiration rate and metabolism in the exposed clams. Since clams are highly consumed shellfish, the consequences for higher tissue bioaccumulation of anthropogenic chemicals to final consumers should be alerted not only at present conditions but more significantly under predicted climatic conditions for humans but also for other components of the marine trophic chain.
The concentration of Ag, As, Cd, Cu, Cr, Hg, Ni, Pb, Se, U, V, and Zn was analyzed in water, sediment, and different organs of Prochilodus lineatus (muscle, liver and gill) in three commercially important catch areas along La Plata Basin, namely High Paraná River (HPR), Middle Paraná River (MPR) and Río de la Plata Estuary (RPE). The concentration of As, Cu and Zn (RPE) and Se (HPR) in water and As (RPE), Ni (HPR and MPR), Cu and Cr (all sites) in sediments exceeded the limits considered as hazardous for aquatic life. With the exception of Se (sediment-liver) and Pb (sediment-liver and sediment-gill), it was not observed a significant correlation between the element concentration in tissues and that found in water and sediment. No correlation was found between the size fish and element concentrations. Liver appeared to be the main storage tissue of trace elements and was classified as a macroconcentrator of Ag, Cu, Hg and Zn. Levels of trace elements in muscle of streaked prochilod were below recommended maximum levels established by national and international guidelines. According to Target Hazard Quotient (THQ), the consumption of muscle of P. lineatus from three sampling sites did not present toxicological risk for general and fishermen populations. Multivariate analyses suggest that the three groups studied remain with an important degree of geographical segregation, indicating that regulations should be revised according to the presence of contaminants in the different fishing areas.
Nineteen elements were analyzed by ICP-MS in surface water from 44 sampling stations from La Plata Basin (South America), including 5 countries and 7 different environments (streams, rivers, meanders, flood valleys, wetland, dam, delta and estuary). A multi-elemental pollution index (EPI) was employed for classifying the water quality while, the potential non-carcinogenic risk through direct ingestion and skin contact were assessed using Hazard Quotients (HQ). Except Fe (2.5-692 μg/L), elements were in accordance to the quality standards for drinking and biota aquatic protection set by international guidelines. EPI indicated a critical scenario for 10 sampling sites (dam, estuary and some meanders, rivers and streams), while the remaining 34 sites were classified within the high pollution status. HQ indicated that water would not be safe (HQ=1.0-2.11) for human consumption in 5 areas (wetland and river and estuary) while, results indicated that there would be no risk (HQ=0.0005-0.01) through chronic water contact. The main contributors to the HQ indices were As (up to 6.3 µg/L) and Li (up to 49.9 µg/L). Correlations (r≥0.8) such as Li-B, Li-As, Li-Sr, Li-Ba, Li-U, Li-conductivity, B-As, B-Sr, B-Ba, B-U, B-conductivity, Sr-Ba, Sr-U, Sr-conductivity provided powerful information about natural sources of some elements. Principal component analysis showed relations between elements and environments such as As, Li, Cs with Andes Mountains and Al and Fe with Atlantic forest. Results allowed to propose management policy recommendations based on strategies for reducing the direct water ingestion, removal methods, and implementation of monitoring programs in the most affected water bodies.
Emissions of metals and metalloids (Hg; Cd; Cr; Cu; Pb; Ni; Zn; Fe; Mn; As; Se) generated by natural (e.g.,geothermal activity) or anthropic causes (eg., industry or mining) represent a worldwide contamination problem, especially in developing countries. Exposure to high concentrations of these elements is harmful to living beings, including humans. Information on this type of contamination is scarce and fragmented, limiting research which could benefit from these data. To know the state of the research, we reviewed the studies of environmental pollution by metals and metalloids carried out on animal species in Latin America. The use of animals as biomonitors of contamination by metals and metalloids is a continuously expanding practice that allows for early detection of problems. With this work, we were able to identify the most studied areas in Latin America(Amazon, Gulf of California, coastal area between Rio de Janeiro and Florianopolis and River Plate Estuary). Moreover, we provide information on the most studied metals (Hg, Cd, Cu, Pb, Zn) and wild species, which evidence the use of endangered species. The data reviewed should help researchers to direct their efforts towards sparsely researched areas and facilitate bibliographic consultation of scientific information on exposure to metals and metalloids in Latin America.
Mussels, such as the marine bivalve Mytilus galloprovincialis are sentinels for marine pollution but they are also excellent bioindicators under laboratory conditions. For that, in this study we tested the modulation of biochemical responses under realistic concentrations of the toxic metal Lead (Pb) in water for 28 days under different conditions of salinity and temperature, including control condition (temperature 17 ± 1.0 °C and salinity 30 ± 1.0) as well as those within the range expected to occur due to climate change predictions (± 5 in salinity and + 4 °C in temperature). A comprehensive set of biomarkers was applied to search on modulation of biochemical responses in terms of energy metabolism, energy reserves, oxidative stress and damage occurrence in lipids, proteins as well as neurotoxicity signs. The application of an integrative Principal Coordinates Ordination (PCO) tool was successful and demonstrated that Pb caused an increase in the detoxification activity mainly evidenced by glutathione S-transferases and that the salinities 25 and 35 were, even in un-exposed mussels, responsible for cell damage seen as increased levels of lipid peroxidation (at salinity 25) and oxidised proteins (at salinity 35).
This study evaluated the Limnoperna fortunei (golden mussel) as a bioindicator of cytotoxicity and genotoxicity in aquatic environments contaminated by heavy metals. Five groups of 50 subjects each were exposed to different concentration of mercuric chloride (HgCl2) (0.001 mg/L, group I; 0.005 mg/L, group II; 0.01 mg/L, group II; 0.02 mg/L, group IV; and 0.1 mg/L, group V). The control group for both chronic and acute treatment did not receive HgCl2. For chronic exposure, the respective groups were placed in aquaria with water contaminated with the above concentrations of HgCl2. For acute exposure, the different concentrations of HgCl2 were injected into the posterior adductor muscle of the individuals belonging to the aforementioned groups. The biological matrix used in the tests was the whole body muscle. Tests (cell viability assay, alkaline comet test; enumeration of micronuclei and necrotic cells, quantification of Hg content in tissues and water, and histopathological analysis of tissues), were carried out on the 7th, 15th, and 30th treatment days or 2 h after injection. Our results demonstrated that L. fortunei showed cell damage in both chronic and acute exposure groups. Significant DNA damage was observed at both the 15th (0.1 mg/L) and 30th (0.01–0.1 mg/L) days of chronic exposure. However, in acute treatment all concentrations induced DNA breaks. The presence of necrosis increased at all concentrations tested for both acute and chronic exposure. Tissue mercury retention on the 15th day was higher than on the 30th day of exposure, while in the same period, there was a decrease in the mercury content of aquarium water. Taking the data together, it is concluded that L. fortunei as a possible bioindicator of the quality of aquatic environments.
Biomarkers (CAT, SOD, GPx, and AChE) were investigated in hepatopancreas and soft tissues of mussels (Mytilus galloprovincialis) and clams (Tapes decussatus) in response to metal and pesticide pollutions in Homa Lagoon. Mussel and clam samples were seasonally collected in January, May, August and November 2015. According to the results, higher metal concentrations determined in the soft tissues of both species in January. Hepatopancreas was found to accumulate higher levels of metals than soft tissue. Metal deposition potential in clams were higher than mussels. Pesticide levels were below detection limits in clams. In mussels, only 4,4′-DDE was detected in soft tissues and hepatopancreas in November. Highest AChE, CAT, SOD and GPx activities in mussels were determined in January. On the other hand, maximum AChE and CAT activities in clams were found in November. Seasonal variations of metals and biomarker levels might be related to species-specific physiology and environmental conditions.
Interest on the effects of emerging contaminants over aquatic organisms has increased in the last years. Nonetheless, the toxic action of classical natural and anthropogenically-driven metals has also to be monitored, especially because they reflect real environmental situations. For that, in the present study we focused on the effects on the marine mussel Mytilus galloprovincialis of the personal care product Triclosan (TCS) and Lead (Pb), as toxic metal, under separate and co-exposure situations at environmentally relevant concentrations: TCS (1 μg/L) and Pb (50 μg/L). The consideration of an additional factor such as an increase in ambient temperature was also included to provide a forecasted scenario of climate change: from the ambient temperature at actual conditions (17 °C) to a predicted warming situation (22 °C). Water chemical characterization and some physical properties and bioaccumulation of TCS and Pb in mussels at the end of the experiment (28 days) was considered. The parameters followed up comprise the energy related system production (electron transport system) and glycogen and protein reserves. Antioxidant enzymatic defences towards reactive oxygen species (ROS) and the consequences of ROS damage over endogenous lipids (LPO) and proteins (PC). Overall the results suggested only particular responses to chemical exposures at 17 °C whereas at 22 °C the detoxification machinery was set up and this prevented the occurrence of LPO. Nonetheless, PC formation occurred under Pb and TCS + Pb co-exposure at the highest temperature. Due to the complexity of the study: 4 chemical conditions, 2 temperatures and 10 biomarkers considered, a principal component ordination (PCO) analysis was included. The results of this integrative analysis confirmed a clear effect of the temperature, more responsiveness to drugs at 22 °C and in all likelihood due to Pb presence.
Determination of relative sensitivity of biota following exposures to contaminants including metals is important for environmental protection. Copper (Cu), although biologically essential can be highly toxic to biota if present at higher concentrations in the natural environment. Given its ubiquitous presence within coastal and inland water bodies, we compared Cu-induced genotoxicity in two ecologically important mussel species, the freshwater Dreissena polymorpha (DP) and marine Mytilus galloprovincialis (MG), along with its tissue specific accumulation. Novel biomarker in terms of induction of gamma H2AX (γ-H2AX) foci, along with comet assay and induction of micronuclei (MN) were used to determine DNA damage response (DDR) in these two species following exposure to a range of Cu concentrations (18, 32, 56 μg L⁻¹) for 10 days. Concentration-dependent increases in Cu concentration in gill tissue, as determined by Inductively Coupled Plasma Mass Spectrometry (ICP-MS), were paralleled by a greater degree of genotoxicity. An induction of γ-H2AX foci was present in all Cu exposure concentrations, proving this technique to be a sensitive and suitable biomarker of genotoxicity in bivalves. The multi-biomarker approach adopted here suggests firstly that in parallel with MG, which is widely used to assess the health of marine and coastal environment, DP is also suitable representative of inland water bodies, and that there is a similar mechanism of action for the induction of genotoxicity between the two species, following exposure to Cu. Secondly, for genotoxicity assessment a battery of responses could simultaneously be assessed in these two bivalve species. Finally, for adequate protection of the environment it is vital to adopt a multi-biomarker, multi-species approach to determine adverse biological effects to gain a holistic understanding of the real threat posed by contaminants to hydrosphere.
This work aims to assess the effects induced by 24 h exposure to a sub‐toxic copper concentration on reproductive system (gonads, spermatozoa and protamine‐like proteins) of Mytilus galloprovincialis. Inductively coupled plasma‐mass spectrometry indicated accumulation of this metal in gonads, spermatozoa and protamine‐like proteins of exposed mussels. Further, RT‐qPCR analyses showed altered expression levels of mt10 and protamine‐like proteins genes in spermatozoa and gonads, respectively of exposed mussels. Protamine‐like proteins, which represent the main basic component of sperm chromatin of this organism, showed, in exposed mussels, an higher DNA binding affinity and a different DNA binding mode. Moreover, an increased amount of NaCl was required for the release from sperm nuclei of PL‐III, the main protamine‐like proteins component. Finally, protamine‐like proteins, extracted from exposed mussels promoted DNA oxidative damage, in the presence of H2O2. These results demonstrate that also tolerable copper amount could affect protamine‐like proteins properties and determine negative effects on reproductive system of this organism. These analyses could be useful to develop quick and efficient chromatin‐based genotoxicity tests for pollution biomonitoring programs.
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Climate change and pollution resulting from human activities in the Arctic require reliable monitoring systems in sentinel species. Mytilus spp. are used as sentinel species all around the world. The use of Mytilus spp. in environmental monitoring requires knowledge about natural variations in pollution biomarkers. Seasonal variations in baseline levels of biomarkers were studied over a year in the mussels from both upper and lower littoral zones in Rakkfjorden, Norway, as they underwent their annual reproductive cycle. Spatial variations of these biomarker baseline levels were measured in five mussel populations within a 60-km radius from Rakkfjorden to investigate universality of the results from the specific population of Rakkfjorden at a regional scale. Seasonal variations in biomarker baseline levels were revealed and seemed to be related to the reproductive state of the mussels and the tidal zone. The mussels appeared to be more sensitive to oxidative stress during gametogenesis in autumn and winter, when having lower lysosome membrane stability and lower baseline levels of antioxidant biomarkers. An increase in baseline levels of these biomarkers was reported during spawning in spring, however, it was not possible to reveal whether these changes were due to spawning, or to a higher metabolic activity in mussels in response to elevated water temperature and food abundance. The differences between the tidal zones were reflected in reduced size of the mussels from the upper littoral zone, their late spawning in the season and increased baseline levels of antioxidant biomarkers during the coldest month, indicating a more challenging environment in the upper littoral zone. The spatial study indicated that the biomarker baseline levels measured in Rakkfjorden were no different from the levels measured in the mussels from five other sites and thus, are representative for mussels on a regional scale.
We investigated the impacts of acute cold and heat exposure on mortality, oxidative stress, antioxidative and stress response, and apoptotic activation in gill tissue of Mytilus galloprovincialis. Our results showed that both cold (4 °C) and heat stress (35 °C) resulted in high mortality. Reactive oxygen species increased significantly after cold stress at 4 °C and heat stress at 30 °C (P < 0.01). In addition, both malondialdehyde content and antioxidative enzyme (SOD, CAT and GSH-Px) activities increased significantly after both cold and heat stress (P < 0.05). However, the transcripts of Hsp70 up-regulated only after heat stress. Western-blot analysis of gill extracts showed that the phosphorylation levels of p38-MAPK were enhanced only at the beginning of cold and heat stress, but elevated activation levels of caspase-3 were observed throughout a 24-h recovery period. Our results suggested that extreme cold and heat air exposure resulted in oxidative stress in the gill tissue of the mussels. The antioxidant enzymes are crucial under both cold and heat exposure, while Hsp70 is induced by heat stress. It seems that p38-MAPK plays a key role in the early response to both cold and heat stress and that apoptosis might be activated in the gill tissue of M. galloprovincialis.
Although some studies have showed the effects of different crystalline structures of nTiO2 (anatase and rutile) and their applicability in several fields, few studies has analyzed the effect of coexposure with other environmental contaminants such as copper. Thus, the objective of this study was to evaluate if the coexposure to nTiO2 (nominal concentration of 1 mg/L; anatase or rutile) can increase the incorporation and toxic effect induced by Cu (nominal concentration of 56 μg/L) in different tissues of Linmoperna fortunei after 120 h of exposure. Our results showed that the coexposure increased the accumulation of Cu in the gills and adductor muscle independently of the crystalline form and can positively or negatively modulate the antioxidant system, depending on the tissue analyzed. However, exposure only to rutile nTiO2 induced damage in the adductor muscle evidenced by the infiltration of hemocytes in this tissue. Additionally, histomorphometric changes based on fractal dimension analysis showed that coexposure to both forms of nTiO2 induced damage in the same tissue. These results suggest that both crystalline forms exhibited toxicity depending on the analyzed tissue and that coexposure of nTiO2 with Cu may be harmful in L. fortunei, indicating that increased attention to the use and release of nTiO2 in the environment is needed to avoid deleterious effects in aquatic biota.
As a consequence of climate change, global warming is expected to increase during the 21st century. Taking this into account, the impact of rising temperatures on the native R. decussatus and introduced R. philippinarum bivalve species was assessed, through biochemical and mRNA expression analyses. Our findings showed that at 21 °C, the electron transport system and antioxidant enzymes activities, as well as the expression of Hsp70 gene were induced in R. decussatus when compared with 17 °C. On the other hand, at 25 °C, results suggested that R. decussatus closed their valves during short periods, probably as a behavioral strategy, downregulating the expression of genes associated with mitochondrial metabolism (Cox-1 and 16S) and molecular chaperone (Hsp70) compared with organisms at 17 °C. In addition, the introduced species (R. philippinarum) increased the electron transport system and antioxidant activities, as well as gene expression of antioxidant enzymes and molecular chaperone (Hsp70) at 21 °C. However, antioxidant mechanisms may not have been enough to prevent together lipid membrane damages at 21 °C. At 25 °C, R. philippinarum presented increased electron transport system and antioxidant mechanisms, as well as the expression of genes associated with apoptosis regulation and molecular chaperone. Overall, the present findings indicate that in a global warming scenario both species are able to induce different mechanisms to mitigate the impacts of temperature increase.