Browsing by Author "Carvalho, Ricardo Cruz de"
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- Effect biomarkers of the widespread antimicrobial triclosan in a marine model diatomPublication . Duarte, Bernardo; Feijão, Eduardo; Carvalho, Ricardo Cruz de; Matos, Ana Rita; Cabrita, Maria Teresa; Novais, Sara C.; Moutinho, Ariana; Lemos, Marco F.L.; Marques, João Carlos; Caçador, Isabel; Reis-Santos, Patrick; Fonseca, VanessaThe present-day COVID-19 pandemic has led to the increasing daily use of antimicrobials worldwide. Triclosan is a manmade disinfectant chemical used in several consumer healthcare products, and thus frequently detected in surface waters. In the present work, we aimed to evaluate the effect of triclosan on diatom cell photophysiology, fatty acid profiles, and oxidative stress biomarkers, using the diatom Phaeodactylum tricornutum as a model organism. Several photochemical effects were observed, such as the lower ability of the photosystems to efficiently trap light energy. A severe depletion of fucoxanthin under triclosan application was also evident, pointing to potential use of carotenoid as reactive oxygen species scavengers. It was also observed an evident favouring of the peroxidase activity to detriment of the SOD activity, indicating that superoxide anion is not efficiently metabolized. High triclosan exposure induced high cellular energy allocation, directly linked with an increase in the energy assigned to vital functions, enabling cells to maintain the growth rates upon triclosan exposure. Oxidative stress traits were found to be the most efficient biomarkers as promising tools for triclosan ecotoxicological assessments. Overall, the increasing use of triclosan will lead to significant effects on the diatom photochemical and oxidative stress levels, compromising key roles of diatoms in the marine system.
- Effects of glyphosate-based herbicide on primary production and physiological fitness of the macroalgae Ulva lactucaPublication . Carvalho, Ricardo Cruz de; Feijão, Eduardo; Matos, Ana Rita; Cabrita, Maria Teresa; Utkin, Andrei B.; Novais, Sara C.; Lemos, Marco F.L.; Caçador, Isabel; Marques, João Carlos; Reis-Santos, Patrick; Fonseca, Vanessa F.; Duarte, BernardoThe use of glyphosate-based herbicides (GBHs) worldwide has increased exponentially over the last two decades increasing the environmental risk to marine and coastal habitats. The present study investigated the effects of GBHs at environmentally relevant concentrations (0, 10, 50, 100, 250, and 500 ug.L-1) on the physiology and biochemistry (photosynthesis, pigment, and lipid composition, antioxidative systems and energy balance) of Ulva lactuca, a cosmopolitan marine macroalgae species. Although GBHs cause deleterious effects such as the inhibition of photosynthetic activity, particularly at 250 ug.L-1, due to the impairment of the electron transport in the chloroplasts, these changes are almost completely reverted at the highest concentration (500 ug.L-1). This could be related to the induction of tolerance mechanisms at a certain threshold or tipping point. While no changes occurred in the energy balance, an increase in the pigment antheraxanthin is observed jointly with an increase in ascorbate peroxidase activity. These mechanisms might have contributed to protecting thylakoids against excess radiation and the increase in reactive oxygen species, associated with stress conditions, as no increase in lipid peroxidation products was observed. Furthermore, changes in the fatty acids profile, usually attributed to the induction of plant stress response mechanisms, demonstrated the high resilience of this macroalgae. Notably, the application of bio-optical tools in ecotoxicology, such as pulse amplitude modulated (PAM) fluorometry and laser-induced fluorescence (LIF), allowed separation of the control samples and those treated by GBHs in different concentrations with a high degree of accuracy, with PAM more accurate in identifying the different treatments.
- Effects of propranolol on growth, lipids and energy metabolism and oxidative stress response of Phaeodactylum tricornutumPublication . Duarte, Bernardo; Feijão, Eduardo; Carvalho, Ricardo Cruz de; Duarte, Irina A.; Silva, Marisa; Matos, Ana Rita; Cabrita, Maria Teresa; Novais, Sara C.; Lemos, Marco F.L.; Marques, João Carlos; Caçador, Isabel; Reis-Santos, Pactick; Fonseca, Vanessa F.Present demographic trends suggest a rise in the contributions of human pharmaceuticals into coastal ecosystems, underpinning an increasing demand to evaluate the ecotoxicological effects and implications of drug residues in marine risk assessments. Propranolol, a non-selective B-adrenoceptor blocker, is used worldwide to treat high blood pressure conditions and other related cardiovascular conditions. Although diatoms lack B-adrenoceptors, this microalgal group presents receptor-like kinases and proteins with a functional analogy to the animal receptors and that can be targeted by propranolol. In the present work, the authors evaluated the effect of this non-selective B-adrenoceptor blocker in diatom cells using P. tricornutum as a model organism, to evaluate the potential effect of this compound in cell physiology (growth, lipids and energy metabolism and oxidative stress) and its potential relevance for marine ecosystems. Propranolol exposure leads to a significant reduction in diatom cell growth, more evident in the highest concentrations tested. This is likely due to the observed impairment of the main primary photochemistry processes and the enhancement of the mitochondrial respiratory activity. More specifically, propranolol decreased the energy transduction from photosystem II (PSII) to the electron transport chain, leading to an increase in oxidative stress levels. Cells exposed to propranolol also exhibited high-dissipated energy flux, indicating that this excessive energy is effciently diverted, to some extent, from the photosystems, acting to prevent irreversible photoinhibition. As energy production is impaired at the PSII donor side, preventing energy production through the electron transport chain, diatoms appear to be consuming storage lipids as an energy backup system, to maintain essential cellular functions. This consumption will be attained by an increase in respiratory activity. Considering the primary oxygen production and consumption pathways, propranolol showed a significant reduction of the autotrophic O2 production and an increase in the heterotrophic mitochondrial respiration. Both mechanisms can have negative effects on marine trophic webs, due to a decrease in the energetic input from marine primary producers and a simultaneous oxygen production decrease for heterotrophic species. In ecotoxicological terms, bio-optical and fatty acid data appear as highly effcient tools for ecotoxicity assessment, with an overall high degree of classification when these traits are used to build a toxicological profile, instead of individually assessed.
- Fluoxetine arrests growth of the model diatom Phaeodactylum tricornutum by increasing oxidative stress and altering energetic and lipid metabolismPublication . Feijão, Eduardo; Carvalho, Ricardo Cruz de; Duarte, Irina A.; Matos, Ana Rita; Cabrita, Maria Teresa; Novais, Sara C.; Lemos, Marco F.L.; Caçador, Isabel; Marques, João Carlos; Reis-Santos, Patrick; Fonseca, Vanessa F.; Duarte, BernardoPharmaceutical residues impose a new and emerging threat to aquatic environments and its biota. One of the most commonly prescribed pharmaceuticals is the antidepressant fluoxetine, a selective serotonin re-uptake inhibitor that has been frequently detected, in concentrations up to 40 ug L-1, in aquatic ecosystems. The present study aims to investigate the ecotoxicity of fluoxetine at environmentally relevant concentrations (0.3, 0.6, 20, 40, and 80 ug L-1) on cell energy and lipid metabolism, as well as oxidative stress biomarkers in the model diatom Phaeodactylum tricornutum. Exposure to higher concentrations of fluoxetine negatively affected cell density and photosynthesis through a decrease in the active PSII reaction centers. Stress response mechanisms, like b-carotene (b-car) production and antioxidant enzymes [superoxide dismutase (SOD) and ascorbate peroxidase (APX)] up-regulation were triggered, likely as a positive feedback mechanism toward formation of fluoxetine-induced reactive oxygen species. Lipid peroxidation products increased greatly at the highest fluoxetine concentration whereas no variation in the relative amounts of long chain polyunsaturated fatty acids (LC-PUFAs) was observed. However, monogalactosyldiacylglycerol-characteristic fatty acids such as C16:2 and C16:3 increased, suggesting an interaction between light harvesting pigments, lipid environment, and photosynthesis stabilization. Using a canonical multivariate analysis, it was possible to evaluate the efficiency of the application of bio-optical and biochemical techniques as potential fluoxetine exposure biomarkers in P. tricornutum. An overall classification efficiency to the different levels of fluoxetine exposure of 61.1 and 88.9% were obtained for bio-optical and fatty acids profiles, respectively, with different resolution degrees highlighting these parameters as potential efficient biomarkers. Additionally, the negative impact of this pharmaceutical molecule on the primary productivity is also evident alongside with an increase in respiratory oxygen consumption. From the ecological point of view, reduction in diatom biomass due to continued exposure to fluoxetine may severely impact estuarine and coastal trophic webs, by both a reduction in oxygen primary productivity and reduced availability of key fatty acids to the dependent heterotrophic upper levels.
- Fluoxetine induces photochemistry-derived oxidative stress on Ulva lactucaPublication . Feijão, Eduardo; Carvalho, Ricardo Cruz de; Duarte, Irina A.; Matos, Ana Rita; Cabrita, Maria Teresa; Utkin, Andrei B.; Caçador, Isabel; Marques, João Carlos; Novais, Sara C.; Lemos, Marco F.L.; Reis-Santos, Patrick; Fonseca, Vanessa; Duarte, BernardoEmerging pollutants impose a high degree of stress on marine ecosystems, compromising valuable resources, the planet and human health. Pharmaceutical residues often reachmarine ecosystems, and their input is directly related to human activities. Fluoxetine is an antidepressant, and one of the most prescribed selective serotonin reuptake inhibitors globally and has been detected in aquatic ecosystems in concentrations up to 40 μg L−1. The present study aims to evaluate the impact of fluoxetine ecotoxicity on the photochemistry, energy metabolism and enzyme activity of Ulva lactuca exposed to environmentally relevant concentrations (0.3, 0.6, 20, 40, and 80 μg L−1). Exogenous fluoxetine exposure induced negative impacts on U. lactuca photochemistry, namely on photosystem II antennae grouping and energy fluxes. These impacts included increased oxidative stress and elevated enzymatic activity of ascorbate peroxidase and glutathione reductase. Lipid content increased and the altered levels of key fatty acids such as hexadecadienoic (C16:2) and linoleic (C18:2) acids revealed strong correlations with fluoxetine concentrations tested. Multivariate analyses reinforced the oxidative stress and chlorophyll a fluorescence-derived traits as efficient biomarkers for future toxicology studies.
- Glyphosate-based herbicide toxicophenomics in marine diatoms: impacts on primary production and physiological fitnessPublication . Carvalho, Ricardo Cruz de; Feijão, Eduardo; Matos, Ana Rita; Cabrita, Maria Teresa; Novais, Sara C.; Lemos, Marco F.L.; Caçador, Isabel; Marques, João Carlos; Reis-Santos, Patrick; Fonseca, Vanessa F.; Duarte, BernardoGlyphosate is the main active component of the commercial formulation Roundup®, the most widely used chemical herbicide worldwide. However, its potential high toxicity to the environment and throughout trophic webs has come under increasing scrutiny. The present study aims to investigate the application of bio-optical techniques and their correlation to physiological and biochemical processes, including primary productivity, oxidative stress, energy balance, and alterations in pigment and lipid composition in Phaeodactylum tricornutum, a representative species of marine diatoms, using the case study of its response to the herbicide glyphosate-based Roundup® formulation, at environmentally relevant concentrations. Cultures were exposed to the herbicide formulation representing effective glyphosate concentrations of 0, 10, 50, 100, 250, and 500 ug L-1. Results showed that high concentrations decreased cell density; furthermore, the inhibition of photosynthetic activity was not only caused by the impairment of electron transport in the thylakoids, but also by a decrease of antioxidant capacity and increased lipid peroxidation. Nevertheless, concentrations of one of the plastidial marker fatty acids had a positive correlation with the highest concentration as well as an increase in total protein. Cell energy allocation also increased with concentration, relative to control and the lowest concentration, although culture growth was inhibited. Pigment composition and fatty acid profiles proved to be efficient biomarkers for the highest glyphosate-based herbicide concentrations, while bio-optical data separated controls from intermediate concentrations and high concentrations.
- Potential of Asparagopsis armata as a biopesticide for weed control under an invasive seaweed circular-economy frameworkPublication . Duarte, Bernardo; Carreiras, João; Feijão, Eduardo; Carvalho, Ricardo Cruz de; Matos, Ana Rita; Fonseca, Vanessa F.; Novais, Sara C.; Lemos, Marco F. L.Marine macroalgae have been increasingly targeted as a source of bioactive compounds to be used in several areas, such as biopesticides. When harvesting invasive species, such as Asparagopsis armata, for this purpose, there is a two-folded opportunity: acquiring these biomolecules from a low-cost resource and controlling its spreading and impacts. The secondary metabolites in this seaweed’s exudate have been shown to significantly impact the physiology of species in the ecosystems where it invades, indicating a possible biocidal potential. Considering this in the present work, an A. armata exudate cocktail was applied in the model weed Thellungiella halophila to evaluate its physiological impact and mode of action, addressing its potential use as a natural biocide. A. armata greatly affected the test plants’ physiology, namely, their photochemical energy transduction pathway (impairing light-harvesting and chemical energy production throughout the chloroplast electron transport chain), carotenoid metabolism and oxidative stress. These mechanisms of action are similar to the ones triggered when using the common chemical pesticides, highlighting the potential of the A. armata exudate cocktail as an eco-friendly biopesticide.