Posts Tagged 'toxicants'

Seawater acidification and emerging contaminants: a dangerous marriage for haemocytes of marine bivalves


• Reduced pH and diclofenac affect haemocyte parameters in exposed mussels and clams.

• During exposure effects of pH persisted, those of diclofenac appeared later.

• Different patterns in haemocyte responses were observed in the two species.

• Interaction between pH and diclofenac was more evident in mussels.


The combined effects of seawater acidification and the non-steroidal anti-inflammatory drug diclofenac on haemocyte parameters of the mussel Mytilus galloprovincialis and the clam Ruditapes philippinarum were investigated for the first time. Animals were maintained for one week (T0) in natural pH condition (8.1) and two reduced pH values (pH −0.4 units and pH −0.7 units). Bivalves were then exposed for additional 14 days (T1 and T2) to the three experimental pH values in both the presence and absence of environmentally realistic concentrations of diclofenac (0.05 and 0.50 μg/L). To assess potential impairment in immunosurveillance, haemocyte parameters (total haemocyte count, haemocyte volume and diameter, Neutral Red uptake, haemocyte proliferation and lysozyme activity) were measured after 7, 14 and 21 days of exposure to differing pH value or pH/diclofenac combinations. In both species, pH affected the whole haemocyte data set at all sampling times, influencing most of the parameters measured at T0 and T1 in clams, and at T2 in mussels. Conversely, in both species diclofenac affected the overall haemocyte response at T2 only. However, in R. philippinarum a higher number of haemocyte parameters were significantly influenced even at T1. A significant interaction between pH and diclofenac was mainly evident in mussels, affecting haemocyte size and lysozyme activity at both T1 and T2. Overall, the results obtained demonstrated that the experimental conditions tested can alter markedly haemocyte parameters in marine bivalves.

Continue reading ‘Seawater acidification and emerging contaminants: a dangerous marriage for haemocytes of marine bivalves’

Stress responses in Crassostrea gasar exposed to combined effects of acute pH changes and phenanthrene


• Acidification is capable to unbalance transcription of biotransformation genes.

• CYP2AU1, CYP2-like2 and GSTΩ genes were upregulated at pH 6.5.

• Water acidification increases gill’s susceptibility to oxidative stress.

• PHE activated enzymatic antioxidant system after 96 h exposure.

• Oysters can protect against with oxidative stress.


Ocean acidification is a result of the decrease in the pH of in marine water, caused mainly by the increase in CO2 released in the atmosphere and its consequent dissolution in seawater. These changes can be dramatic for marine organisms especially for oysters Crassostrea gasar if other stressors such as xenobiotics are present. The effect of pH changes (6.5, 7.0 and 8.2) was assessed on the transcript levels of biotransformation [cytochromes P450 (CYP2AU1, CYP2-like2) and glutathione S-transferase (GSTΩ-like)] and antioxidant [superoxide dismutase (SOD-like), catalase (CAT-like) and glutathione peroxidase (GPx-like)] genes, as well as enzyme activities [superoxide dismutase, (SOD), catalase (CAT), glutathione reductase (GR), glutathione-S-transferases (GST) and glucose-6-phosphate dehydrogenase (G6PDH)] and lipid peroxidation (MDA) in the gills of Crassostrea gasar exposed to 100 μg·L−1 of phenanthrene (PHE) for 24 and 96 h. Likewise, the PHE burdens was evaluated in whole soft tissues of exposed oysters. The accumulation of PHE in oysters was independent of pH. However, acidification promoted a significant decrease in the transcript levels of some protective genes (24 h exposure: CYP2AU1 and GSTΩ-like –; 96 h exposure: CAT-like and GPx-like), which was not observed in the presence of PHE. Activities of GST, CAT and SOD enzymes increased in the oysters exposed to PHE at the control pH (8.2), but at a lower pH values, this activation was suppressed, and no changes were observed in the G6PDH activity and MDA levels. Biotransformation genes showed better responses after 24 h, and antioxidant-coding genes after 96 h, along with the activities of antioxidant enzymes (SOD, CAT), probably because biotransformation of PHE increases the generation of reactive oxygen species. The lack of change in MDA levels suggests that antioxidant modulation efficiently prevented oxidative stress. The effect of pH on the responses to PHE exposure should be taken into account before using these and any other genes as potential molecular biomarkers for PHE exposure.

Continue reading ‘Stress responses in Crassostrea gasar exposed to combined effects of acute pH changes and phenanthrene’

Interaction of short-term copper pollution and ocean acidification in seagrass ecosystems: toxicity, bioconcentration and dietary transfer


• Toxicity and bioconcentration of copper in seagrasses were not affected by pH.
• Complex copper-pH interactions were observed in the seagrass photosynthesis.
• Seagrasses can act as a copper source in the food web via direct consumption.


We aimed to show how the predicted pH decrease in the ocean would alter the toxicity, bioconcentration and dietary transfer of trace metal copper on seagrass ecosystems, on a short-term basis. Seagrass Zostera noltei was exposed to two pH levels (8.36 and 8.03) and three copper levels (nominal concentrations, <3, 30 and 300 μg Cu L−1) in a factorial design during 21 days, while Gammarus locusta amphipods were continuously fed with the treated seagrass leaves. We found that the toxicity and bioconcentration of copper in seagrasses were not affected by pH, yet complex copper-pH interactions were observed in the seagrass photosynthesis. We demostrated that seagrasses can act as a copper source in the food web via direct consumption by herbivores. Future research need to investigate the interactive effects on a long-term basis, and to include biochemical and molecular endpoints to provide additional insights to the complex phisiological interactions observed.

Continue reading ‘Interaction of short-term copper pollution and ocean acidification in seagrass ecosystems: toxicity, bioconcentration and dietary transfer’

The effect of pH on the acute toxicity of phenanthrene in a marine microalgae Chlorella salina

Phenanthrene is one of the most abundant polycyclic aromatic hydrocarbons (PAHs) found in continental shelf environment of China and is on the EPA’s Priority Pollutant list. In this study, the effects of phenanthrene on marine algal growth rate were determined after 96-h exposure at pH 6.0, 7.0, 8.0, 9.0, and 10.0 in seawater of salinity 35. Two measuring techniques to assess growth inhibition were also compared using prompt fluorescence and microscopic cell count. The results showed that the toxicity of phenanthrene increased significantly (p < 0.05) with decreasing pH, with the nominal concentration required to inhibit growth rate by 50%, EC50, decreasing from 1.893 to 0.237 mg L−1 as pH decreased from 9.0 to 6.0, with a decrease higher than 55% from 10.0 to 9.0. In addition, the nominal EC50 values calculated in this study were at the same range of some environmental concentrations of phenanthrene close to areas of crude oil exploration. Based on the two measuring techniques, the results showed that cell count and fluorescence measurement were significantly different (p < 0.05), and the nominal EC50 values calculated with cell count measurement were significantly higher than fluorescence measurement at pH 8.0, 9.0 and 10.0. In conclusion, the present studies confirmed that acidification of seawater could affect the toxicity of phenanthrene to this species of microalgae, and which encouraged further studies involving responses of marine organisms to ocean acidification.

Continue reading ‘The effect of pH on the acute toxicity of phenanthrene in a marine microalgae Chlorella salina’

Genotoxic effects of combined multiple stressors on Gammarus locusta haemocytes: Interactions between temperature, pCO2 and the synthetic progestin levonorgestrel


• Combined factors (temperature, pCO2, levonorgestrel) were assessed on G. locusta.

• Comet assay as valid technique to evaluate DNA damage in haemocytes.

• Haemocytes of each treatment were exposed, ex vivo, to an oxidant agent – H2O2.

• Stress factors alone did not significantly increase DNA damage, after 21 days.

• Solvent increased haemocytes DNA damage – not observed in hormone treatments.


Climate change and pharmaceutical contamination are two priority research topics due to their impacts in the aquatic ecosystems and in the food chain structure. In the bottom of many food chains are the invertebrates, like the amphipods, which are important environmental and ecotoxicological models. In this study, we combined the increase of temperature [ambient and warming temperature], pCO2 [normocapnia and hypercapnia] and the synthetic progestin levonorgestrel (LNG) [environmentally relevant concentration (10 ng L−1) and 100-fold higher (1000 ng L−1)] to evaluate the genotoxic effects on the amphipod Gammarus locusta haemocytes, using the comet assay technique. Additionally, the study examined protective/potentiating effects of the three tested factors against hydrogen peroxide (H2O2)-induced DNA damage in haemocytes after ex vivo exposure. Our data revealed no significant effects of any of the three stressors on DNA damage of G. locusta haemocytes or protection against H2O2-induced DNA damage after twenty-one days exposure. Only a significant effect of the solvent was visible, since it was able to induce higher DNA damage (i.e. strand breaks) on exposed individuals. On the other hand, LNG exposure seemed to induce a slight increase of DNA damage after H2O2 exposure. Our findings suggest that more short-term studies to conclude about the genotoxicity and/or protective effects of the stress factors in G. locusta should be made, attending to the fast turnover rate of repairing cells that could have masked impacts seen only after the end of the experiment.

Cruzeiro C., Ramos A., Loganimoce E. M., Arenas F., Rocha E. & Cardoso P. G., 2019. Genotoxic effects of combined multiple stressors on Gammarus locusta haemocytes: Interactions between temperature, pCO2 and the synthetic progestin levonorgestrel. Environmental Pollution 245: 864-872. Article (subscription required).

Effects of oil and global environmental drivers on two keystone marine invertebrates

Ocean warming (OW) and acidification (OA) are key features of global change and are predicted to have negative consequences for marine species and ecosystems. At a smaller scale increasing oil and gas activities at northern high latitudes could lead to greater risk of petroleum pollution, potentially exacerbating the effects of such global stressors. However, knowledge of combined effects is limited. This study employed a scenario-based, collapsed design to investigate the impact of one local acute stressor (North Sea crude oil) and two chronic global drivers (pH for OA and temperature for OW), alone or in combination on aspects of the biology of larval stages of two key invertebrates: the northern shrimp (Pandalus borealis) and the green sea urchin (Strongylocentrotus droebachiensis). Both local and global drivers had negative effects on survival, development and growth of the larval stages. These effects were species- and stage-dependent. No statistical interactions were observed between local and global drivers and the combined effects of the two drivers were approximately equal to the sum of their separate effects. This study highlights the importance of adjusting regulation associated with oil spill prevention to maximize the resilience of marine organisms to predicted future global conditions.

Continue reading ‘Effects of oil and global environmental drivers on two keystone marine invertebrates’

Bioaccumulation and ecotoxicological responses of juvenile white seabream (Diplodus sargus) exposed to triclosan, warming and acidification


• TCS was primarily bioaccumulated by fish muscle, followed by liver and brain.
• Climate change significantly reduced TCS bioaccumulation in all fish tissues.
• TCS and TCS + acidification increased LPO and inhibited HSP70/HSC70 synthesis.
• AChE activity was inhibited by TCS alone or combined with both abiotic stressors.
• VTG content was further diminished by TCS co-exposed with both abiotic stressors.


Triclosan (TCS) is a synthetic microbial compound widely used in the formulation of various personal care products. Its frequent detection in marine ecosystems, along with its physical and chemical properties, suggest that TCS can be highly persistent, being easily bioaccumulated by biota and, therefore, eliciting various toxicological responses. Yet, TCS’s mechanisms of bioaccumulation and toxicity still deserve further research, particularly focusing on the interactive effects with climate change-related stressors (e.g. warming and acidification), as both TCS chemical behaviour and marine species metabolism/physiology can be strongly influenced by the surrounding abiotic conditions. Hence, the aim of this study was to assess TCS bioaccumulation and ecotoxicological effects (i.e. animal fitness indexes, antioxidant activity, protein chaperoning and degradation, neurotoxicity and endocrine disruption) in three tissues (i.e. brain, liver and muscle) of juvenile Diplodus sargus exposed to the interactive effects of TCS dietary exposure (15.9 μg kg−1 dw), seawater warming (ΔTºC = +5 °C) and acidification (ΔpCO2 ∼ +1000 μatm, equivalent to ΔpH = −0.4 units). Muscle was the primary organ of TCS bioaccumulation, and climate change stressors, particularly warming, significantly reduced TCS bioaccumulation in all fish tissues. Furthermore, the negative ecotoxicological responses elicited by TCS were significantly altered by the co-exposure to acidification and/or warming, through either the enhancement (e.g. vitellogenin content) or counteraction/inhibition (e.g. heat shock proteins HSP70/HSC70 content) of molecular biomarker responses, with the combination of TCS plus acidification resulting in more severe alterations. Thus, the distinct patterns of TCS tissue bioaccumulation and ecotoxicological responses induced by the different scenarios emphasized the need to further understand the interactive effects between pollutants and abiotic conditions, as such knowledge enables a better estimation and mitigation of the toxicological impacts of climate change in marine ecosystems.

Continue reading ‘Bioaccumulation and ecotoxicological responses of juvenile white seabream (Diplodus sargus) exposed to triclosan, warming and acidification’

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Ocean acidification in the IPCC AR5 WG II

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