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  • Fluoxetine induces photochemistry-derived oxidative stress on Ulva lactuca
    Publication . 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, Bernardo
    Emerging 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.
  • Comfortably numb: Ecotoxicity of the non-steroidal anti-inflammatory drug ibuprofen on Phaeodactylum tricornutum
    Publication . Feijão, Eduardo; Carvalho, Ricardo da Cruz de; Duarte, Irina A.; Matos, Ana Rita; Cabrita, Maria Teresa; Barreiro, Aldo; Lemos, Marco F.L.; Novais, Sara C.; Marques, João Carlos; Caçador, Isabel; Reis-Santos, P.; Silva, Marisa; Fonseca, Vanessa; Duarte, Bernardo
    Emerging pollutants such as pharmaceuticals are continuously released to aquatic environments posing a rising threat to marine ecosystems. Yet, monitoring routines and ecotoxicity data on biota worldwide for these substances are lacking. Non-steroidal anti-inflammatory drugs are among the most prescribed and found pharmaceuticals in aquatic environments. The toxicity effects of environmentally relevant concentrations of ibuprofen on primary productivity, oxidative stress and lipid metabolism of the diatom Phaeodactylum tricornutum were assessed. Diatom cultures were exposed to 0, 0.8, 3, 40, 100 and 300 μg L-1 ibuprofen concentrations, usually found in the vicinity of wastewater treatment plants and coastal environments. Higher concentrations (100 and 300 μg L-1) had a negative impact in P. triconutum growth, inhibiting the chloroplastic energy transduction in the electron transport chain resulting in lower energy reaching the PS I (r2 = - 0.55, p < 0.05). In contrast, the mitochondrial electron transport and available energy increased (r2 = 0.68 and r2 = 0.85, p < 0.05 respectively), mostly due to enhancements in lipid and protein contents as opposed to reduction of carbohydrates. A general up-regulation of the antioxidant enzymes could contributed to alleviate oxidative stress resulting in the decrease of lipid peroxidation products (r2 = 0.77, p < 0.05). Canonical analysis of principal components was performed and successfully discriminated exposure groups, with optical data excelling in classifying samples to different ibuprofen concentrations, being potentially used as environmental indicators. Finally, the identified mild to severe effects of ibuprofen on diatoms are likely to be exacerbated by the sustained use of this drug worldwide, underpinning the urgency of evaluating the impacts of this pharmaceutical on coastal and marine trophic webs.
  • Effect biomarkers of the widespread antimicrobial triclosan in a marine model diatom
    Publication . 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, Vanessa
    The 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 lactuca
    Publication . 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, Bernardo
    The 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.