Mario Sousa Diniz
@fct.unl.pt
Department of Chemistry/ NOVA SCHOOL OF SCIENCE AND TECHNOLOGY
Universidade Nova de Lisboa
EDUCATION
PhD in environmental sciences
RESEARCH INTERESTS
Environmental Toxicology, Biochemistry, Climate change effects, Proteomics, Food safety
Scopus Publications
Scholar Citations
Scholar h-index
Scholar i10-index
Scopus Publications
Isa Marmelo, Marta Dias, Ana Grade, Pedro Pousão-Ferreira, Mário Sousa Diniz, António Marques, and Ana Luísa Maulvault
Frontiers Media SA
The increasing demand for healthy aquatic products has contributed to the rapid growth of aquaculture. Despite the many progresses, over the years, in aquaculture production, animal welfare and reduction of the stress induced by captivity remain a challenge in this sector, and overcoming it is fundamental to improve animal growth performance and resilience to environmental threats (e.g., disease outbreaks and/or climate change impacts). To address this challenge, aquafeeds biofortified with natural ingredients with functional properties, such as macroalgae, have emerged as a promising strategy to enhance fish immunity. The present study aimed at evaluating the potential of Laminaria digitata to improve juvenile gilthead seabream (Sparus aurata) physiological status and resilience, through the modulation of immunomodulatory and antioxidant responses. To explore this hypothesis, four independent treatments were carried out in which fish were fed with different experimental aquafeeds: control feed and biofortified feeds with 1.5%, 3% and 6% of whole L. digitata (incorporated as a dried and powdered ingredient). Fish (n=9) were collected from each treatment upon 30 and 60 days of biofortification in order to assess animal growth and condition indexes, haematological parameters, innate humoral parameters (antiprotease (AP) activity, peroxidase (POD) activity and immunoglobulin M (IgM)) and oxidative stress biomarkers (lipid peroxidation (LPO), catalase (CAT) activity, glutathione S-transferase (GST) activity, superoxide dismutase (SOD) activity and total antioxidant capacity (T-AOC)). Significant differences (α = 0.05) between treatments in biomarker responses were tested through a nested-design ANOVA, followed by Tukey HSD multiple comparisons. Pearson correlations between the different endpoints were also explored. Results indicate that a dietary inclusion of 1.5% L. digitata is the most effective biofortification dose, significantly improving antioxidant responses of S. aurata, by increasing levels of CAT, GST and T-AOC, along with the reduction in LPO levels. Additionally, the inclusion of 1.5% L. digitata enhances immunity by increasing the levels of IgM and POD, while maintaining the growth performance and condition indices of the fish. These findings underscore the potential utilization of L. digitata as a functional feed ingredient in aquaculture with relevant immune and antioxidant attributes.
Inês Rodrigues, Inês João Ferreira, Regina M. B. O. Duarte, and Mário Diniz
MDPI AG
Atmospheric particulate matter (PM) contains numerous constituents, including organic molecules, inorganic ions, and metals, with some of them possessing hazardous properties. Although mainly associated with air pollution, PM can rapidly be transferred from air and land to aquatic ecosystems, and consequently poses a risk to marine biota. The aim of this work was to evaluate how urban atmospheric PM (a standard reference mixture of urban PM, known to contain various organic and inorganic contaminants), suspended in seawater, may cause toxicity in marine organisms. To this purpose, mussels (Mytilus galloprovincialis) were exposed to two concentrations of suspended PM: 5.7 and 11.4 mg/L. After 7, 14, and 21 days, the animals were collected and the gills and digestive gland were analysed for stress biomarkers (CAT, SOD, GPX, GST, MDA, and Ubi). In general, the results show that exposure to different concentrations of PM caused an increase in GST, UBI, and GPx activities compared to their respective controls. The average activities of GST (87.65 ± 30.23 nmol/min/mg of total protein) in the gills of the animals exposed to 11.4 mg/L of PM increased after 21 days of exposure, and the activity of GPx (8.04 ± 3.09 nmol/min/mg of total protein) in the gills increased after 14 days in the animals exposed to 5.7 mg/L of PM. MDA results also provided information on cellular damage, with the most pronounced effects being found in the gills of exposed mussels. This study confirms that mussels are useful as “early warning” indicators of environmental contamination and provides important information on the effects of PM on marine biota.
Janina Leonie Röckner, Vanessa M. Lopes, José Ricardo Paula, Maria Rita Pegado, Martim Costa Seco, Mário Diniz, Tiago Repolho, and Rui Rosa
Springer Science and Business Media LLC
Rita Dias, Michiel A. Daam, Mário Diniz, and Rita Maurício
Elsevier BV
Antonio Cid-Samamed, Miguel Ángel Correa-Duarte, Andrea Mariño-López, and Mário S. Diniz
MDPI AG
The increasing attention that carbon-based nanomaterials have attracted due to their distinctive properties makes them one of the most widely used nanomaterials for industrial purposes. However, their toxicity and environmental effects must be carefully studied, particularly regarding aquatic biota. The implications of these carbon-based nanomaterials on aquatic ecosystems, due to their potential entry or accidental release during manufacturing and treatment processes, need to be studied because their impacts upon living organisms are not fully understood. In this research work, the toxicity of oxidized multi-walled carbon nanotubes (Ox-MWCNTs) was measured using the freshwater bivalve (Corbicula fluminea) after exposure to different concentrations (0, 0.1, 0.2, and 0.5 mg·L−1 Ox-MWCNTs) for 14 days. The oxidized multi-walled carbon nanotubes were analyzed (pH, Raman microscopy, high-resolution electron microscopy, and dynamic light scattering), showing their properties and behavior (size, aggregation state, and structure) in water media. The antioxidant defenses in the organism’s digestive gland and gills were evaluated through measuring oxidative stress enzymes (glutathione-S-transferase, catalase, and superoxide dismutase), lipid peroxidation, and total ubiquitin. The results showed a concentration-dependent response of antioxidant enzymes (CAT and GST) in both tissues (gills and digestive glands) for all exposure periods in bivalves exposed to the different concentrations of oxidized multi-walled carbon nanotubes. Lipid peroxidation (MDA content) showed a variable response with the increase in oxidized multi-walled carbon nanotubes in the gills after 7 and 14 exposure days. Overall, after 14 days, there was an increase in total Ub compared to controls. Overall, the oxidative stress observed after the exposure of Corbicula fluminea to oxidized multi-walled carbon nanotubes indicates that the discharge of these nanomaterials into aquatic ecosystems can affect the biota as well as potentially accumulate in the trophic chain, and may even put human health at risk if they ingest contaminated animals.
Antonio Cid-Samamed and M. S. Diniz
MDPI AG
This short review aims to critically discuss the recent advances in supramolecular chemistry to achieve the aggregation of nanoplastics in aquatic systems. Polymer modification provides a vital tool for designing novel and ad hoc synthesized surfactants with properties tuned for some specific applications (e.g., stimuli-responsive nanomaterial, conducting polymers), mainly to aggregate other polymers from the environment. Far from the typical use of surfactants, which ease the dispersion of insoluble molecules in water media or aid solubilization of insoluble molecules on local media, in this case, nanoarchitectonics serve researchers to design surfactants with a focus on the capture of nanoplastics from the environment. Additionally, monovalent and divalent salt additions aided NPs in coagulating in the aquatic systems. Finally, the latest research on NPs’ removal efficiency on wastewater treatment plant is reviewed to summarize the advances.
Rui Cereja, Joana P. C. Cruz, Joshua Heumüller, Bernardo Vicente, Ana Amorim, Frederico Carvalho, Sara Cabral, Paula Chainho, Ana C. Brito, Inês J. Ferreira,et al.
MDPI AG
Bivalves accumulate toxins produced by microalgae, thus becoming harmful for humans. However, little information is available about their toxicity to the bivalve itself. In the present work, the physiological stress and damage after the ingestion of toxic dinoflagellate species (Gymnodinium catenatum) and a diatom species (Skeletonema marinoi, which is non-toxic to humans but may be to grazers) in the oyster Magallana angulata are evaluated against a control treatment fed with the chlorophyte Tetraselmis sp. Oysters were exposed for two hours to a concentration of 4 × 104 cells/L of G. catenatum and 2 × 107 cells/L of S. marinoi. The biomarkers superoxide dismutase (SOD), catalase (CAT), glutathione S-Transferase, total Ubiquitin (Ubi) and Acetylcholinesterase (AchE) were assessed. The exposure of M. angulata to G. catenatum lead to a reduction in SOD and AchE activity and ubiquitin concentrations when compared to the control treatment. Moreover, it increased CAT activity in the adductor muscle, and maintained its activity in the other tissues tested. This may be related to the combination of reduced metabolism with the deployment of detoxification processes. S. marinoi also lead to a decrease in all biomarkers tested in the gills and digestive glands. Therefore, both species tested caused physiological alterations in M. angulata after two hours of exposure.
Sara Valente, Filipe Oliveira, Inês João Ferreira, Alexandre Paiva, Rita G. Sobral, Mário S. Diniz, Susana P. Gaudêncio, and Ana Rita Cruz Duarte
American Chemical Society (ACS)
Marine biofouling negatively impacts industries with off-shore infrastructures, such as naval, oil, and aquaculture. To date, there are no ideal sustainable, economic, and environmentally benign solutions to deal with this phenomenon. The advances achieved in green solvents, as well as its application in different industries, such as pharmaceutical and biotechnology, have promoted the emergence of deep eutectic systems (DES). These eutectic systems have applications in various fields and can be revolutionary in the marine-based industrial sector. In this study, the main objective was to investigate the potential use of hydrophobic DES (HDES) based on menthol and natural organic acids for their use as marine antifouling coatings. Our strategy encompassed the physicochemical characterization of different formulations, which allowed us to identify the most appropriate molar ratio and intermolecular interactions for HDES formations. The miscibility of the resulting HDES with the marine coating has been evaluated and proven to be successful. The Men/OL (1:1) system proved to be the most promising in terms of cost-production and thus was the one used in subsequent antifouling tests. The cytotoxicity of this HDES was evaluated using an in vitro cell model (HaCat cells) showing no significant toxicity. Furthermore, the application of this system incorporated into coatings that are used in marine structures was also studied using marine species (Mytilus edulis mussels and Patella vulgata limpets) to evaluate both their antifouling and ecotoxicity effects. HDES Men/OL (1:1) incorporated in marine coatings was promising in reducing marine macrofouling and also proved to be effective at the level of microfouling without viability impairment of the tested marine species. It was revealed to be more efficient than using copper oxide, metallic copper, or ivermectin as antifouling agents. Biochemical assays performed on marine species showed that this HDES does not induce oxidative stress in the tested species. These results are a strong indication of the potential of this HDES to be sustainable and efficiently used in marine fouling control technologies.
Bruno Moreira-Leite, Rafael Antunes, João Cotas, Nuno Martins, Nuno Costa, João P. Noronha, Paulina Mata, and Mário Diniz
MDPI AG
Conventional conservation techniques such as drying, salting or freezing do not allow for preserving the original characteristics of seaweeds. The present work aims to study the impact of minimal processing, in particular “Modified Atmosphere Packaging” (MAP), on the physicochemical characteristics and food safety of two seaweed species, “laver” (Porphyra umbilicalis) and “sea-lettuce” (Ulva lactuca), stored at 6 °C for 15 days. Different parameters were evaluated using analytical methods, namely the composition of headspace gases, color, texture, microorganisms, and volatile organic compounds (VOCs). The main findings of this study were that the MAP treatment was able to inhibit the respiration rate of minimally processed seaweeds, also preserving their color and texture. There was a remarkable reduction in the microbial load for P. umbilicalis treated under modified and vacuum atmospheres, and U. lactuca exhibited relatively steady values with no notable differences between the treatments and the control. Therefore, during the 15-day study period, both seaweeds met the requirements for food safety. GC–TOF-MS allowed to conclude that both MAP and vacuum treatments were more efficient in maintaining the odor characteristics of U. lactuca compared to P. umbilicalis with no significant differences throughout the storage days. Metabolic responses to diverse sources of abiotic stress seemed to account for most of the changes observed.
Pedro Catalão Moura, Jorge Manuel Fernandes, Mário Sousa Diniz, Viktor Fetter, and Valentina Vassilenko
MDPI AG
The inclusion of seaweeds in daily-consumption food is a worthy-of-attention challenge due to their high nutritional value and potential health benefits. In this way, their composition, organoleptic profile, and toxicity must be assessed. This work focuses on studying the volatile organic compounds (VOCs) emitted by three edible seaweeds, Grateloupia turuturu, Codium tomentosum, and Bifurcaria bifurcata, with the aim of deepening the knowledge regarding their organoleptic profiles. Nine samples of each seaweed were prepared in glass vials, and the emitted headspace was analyzed, for the first time, with a gas chromatography—ion mobility spectrometry device, a highly sensitive technology. By statistically processing the collected data through PCA, it was possible to accurately differentiate the characteristic patterns of the three seaweeds with a total explained variance of 98%. If the data were pre-processed through PLS Regression, the total explained variance increased to 99.36%. The identification of 13 VOCs was accomplished through a developed database of compounds. These outstanding values in addition to the identification of the main emissions of VOCs and the utilization of a never-before-used technology prove the capacity of GC-IMS to differentiate edible seaweeds based solely on their volatile emissions, increase the knowledge regarding their organoleptic profiles, and provide an important step forward in the inclusion of these highly nutritional ingredients in the human diet.
Jaquelino Varela, Sandra Martins, Melanie Court, Catarina Pereira Santos, José Ricardo Paula, Inês João Ferreira, Mário Diniz, Tiago Repolho, and Rui Rosa
MDPI AG
Climate change is leading to the loss of oxygen content in the oceans and endangering the survival of many marine species. Due to sea surface temperature warming and changing circulation, the ocean has become more stratified and is consequently losing its oxygen content. Oviparous elasmobranchs are particularly vulnerable as they lay their eggs in coastal and shallow areas, where they experience significant oscillations in oxygen levels. Here, we investigated the effects of deoxygenation (93% air saturation) and hypoxia (26% air saturation) during a short-term period (six days) on the anti-predator avoidance behavior and physiology (oxidative stress) of small-spotted catshark (Scyliorhinus canicula) embryos. Their survival rate decreased to 88% and 56% under deoxygenation and hypoxia, respectively. The tail beat rates were significantly enhanced in the embryos under hypoxia compared to those exposed to deoxygenation and control conditions, and the freeze response duration showed a significant opposite trend. Yet, at the physiological level, through the analyses of key biomarkers (SOD, CAT, GPx, and GST activities as well as HSP70, Ubiquitin, and MDA levels), we found no evidence of increased oxidative stress and cell damage under hypoxia. Thus, the present findings show that the projected end-of-the-century deoxygenation levels elicit neglectable biological effects on shark embryos. On the other hand, hypoxia causes a high embryo mortality rate. Additionally, hypoxia makes embryos more vulnerable to predators, because the increased tail beat frequency will enhance the release of chemical and physical cues that can be detected by predators. The shortening of the shark freeze response under hypoxia also makes the embryos more prone to predation.
Inês João Ferreira, Alexandre Paiva, Mário Diniz, and Ana Rita Duarte
Springer Science and Business Media LLC
Abstract Deep eutectic systems (DES) have shown increasing popularity in last decade; however, the number of studies on the potential toxicity towards living organisms remains scarce. These studies are of the utmost importance to infer on the claimed non-toxicity and biocompatibility of DES. Most articles published, at this moment, only evaluate the toxicity towards a cell model or in different strains of bacteria. For this purpose, in this work, the effect of two DES (betaine:sorbitol:water 1:1:3 and betaine:glycerol 1:2) and their individual components were evaluated at different concentrations after administered via intraperitoneal injection in zebrafish (Danio rerio). The total antioxidant capacity, lipoperoxidation, and the activity of various enzymes that work in different antioxidant pathways (superoxide dismutase, glutathione peroxidase, catalase, and glutathione S-transferase) were assessed. The results show no significant toxicity within the tested concentrations: up to 5000 µM and 3000 µM, for the assays using the system betaine:sorbitol:water 1:1:3 and for betaine:glycerol 1:2, respectively. The toxicity of individual components was studied up to 1000 µM. Based on the encouraging results that have been obtained, it is safe to conclude that these two deep eutectic systems can be used as the new class of environmentally friendly solvents. Graphical Abstract
João Coelho, Mélanie Court, Eve Otjacques, Vanessa M. Lopes, José Ricardo Paula, Tiago Repolho, Mário Diniz, and Rui Rosa
Springer Science and Business Media LLC
Bruno Miguel Campos, Edgar Ramalho, Isa Marmelo, João Paulo Noronha, Manuel Malfeito-Ferreira, Paulina Mata, and Mário Sousa Diniz
IMR Press
BACKGROUND
The aim of this work was the study of the proximate composition and profile of fatty acids, minerals, and some microbiological aspects of four edible seaweed species (Chondrus crispus, Palmaria palmata, Porphyra sp., and Ulva sp.) available in the Portuguese market for food consumption, and produced in a national Integrated Multi-Trophic System (IMTA).
METHODS
Moisture, ash, and total lipids were determined gravimetrically. Crude protein was analysed by Duma's combustion procedures. The total carbohydrate content was assayed by the phenol/sulphuric acid method. The assessment of the fatty acids methyl esters (FAMEs) was determined through GC-MS. Characterization of elemental analysis was performed by ICP-AES. Different standard microbiological methods were applied for microorganisms. Statistics were performed using the non-parametric Mann-Whitney U test to assess significant differences between samples.
RESULTS
Lipid contents (n = 3) were very low (1.6-2.3%), particularly in Palmaria palmata, and Chondrus crispus (1.6-1.7%). The protein content (n = 4) varied from 14.4% in P. palmata to 23.7% in Porphyra sp. Carbohydrates (n = 3) were the major constituent of most seaweeds (31-34%), except in Porphyra sp., with higher content in proteins than carbohydrates. Regarding the fatty acid content (n = 4), in general, saturated fatty acids (SFAs) were the most abundant followed by polyunsaturated fatty acids (PUFAs) and monounsaturated fatty acids (MUFAs). Among macro and trace elements (n = 3), Chondrus crispus shows the highest average content in Zn (71.1 mg ⋅ kg - 1 D.W.), Palmaria palmata the highest average content in K (124.8 g ⋅ kg - 1 D.W.), Porphyra sp. the highest average content in P (2.1 g ⋅ kg - 1 D.W.), and Ulva sp. the highest average content of Ca (5.5 g ⋅ kg - 1 D.W.), Mg (55.8 g ⋅ kg - 1 D.W.), and Fe (336.3 mg ⋅ kg - 1 D.W.). In general, Na and K were the most abundant elements among analysed seaweed. Additionally, the microbiological results (n = 4) comply with the Portuguese guidelines (subgroup 2D) on the application of general principles of food hygiene in ready-to-eat foods.
CONCLUSIONS
Overall, the results highlight the potential of using these seaweeds as an alternative and sustainable source of elements and bioactive compounds to produce enriched food products with a beneficial potential for human nutrition.
Cátia Figueiredo, Tiago F. Grilo, Rui Oliveira, Inês João Ferreira, Fátima Gil, Clara Lopes, Pedro Brito, Pedro Ré, Miguel Caetano, Mário Diniz,et al.
Elsevier BV
Cátia Figueiredo, Tiago F. Grilo, Rui Oliveira, Inês João Ferreira, Fátima Gil, Clara Lopes, Pedro Brito, Pedro Ré, Miguel Caetano, Mário Diniz,et al.
Elsevier BV
Cátia Figueiredo, Tiago F. Grilo, Rui Oliveira, Inês João Ferreira, Fátima Gil, Clara Lopes, Pedro Brito, Pedro Ré, Miguel Caetano, Mário Diniz,et al.
Elsevier BV
Inês João Ferreira, Liane Meneses, Alexandre Paiva, Mário Diniz, and Ana Rita C. Duarte
Elsevier BV
Patrícia Branco, Mário Diniz, and Helena Albergaria
MDPI AG
Lactic acid bacteria (LAB) and Brettanomyces bruxellensis are the main contaminants of bioethanol fermentations. Those contaminations affect Saccharomyces cerevisiae performance and reduce ethanol yields and productivity, leading to important economic losses. Currently, chemical treatments such as acid washing and/or antibiotics are used to control those contaminants. However, these control measures carry environmental risks, and more environmentally friendly methods are required. Several S. cerevisiae wine strains were found to secrete antimicrobial peptides (AMPs) during alcoholic fermentation that are active against LAB and B. bruxellensis strains. Thus, in the present study, we investigated if the fuel-ethanol commercial starter S. cerevisiae Ethanol Red (ER) also secretes those AMPs and evaluated its biocontrol potential by performing alcoholic fermentations with mixed-cultures of ER and B. bruxellensis strains and growth assays of LAB in ER pre-fermented supernatants. Results showed that all B. bruxellensis strains were significantly inhibited by the presence of ER, although LAB strains were less sensitive to ER fermentation metabolites. Peptides secreted by ER during alcoholic fermentation were purified by gel-filtration chromatography, and a bioactive fraction was analyzed by ELISA and mass spectrometry. Results confirmed that ER secretes the AMPs previously identified. That bioactive fraction was used to determine minimal inhibitory concentrations (MICs) against several LAB and B. bruxellensis strains. MICs of 1–2 mg/mL were found for B. bruxellensis strains and above 2 mg/mL for LAB. Our study demonstrates that the AMPs secreted by ER can be used as a natural preservative in fuel-ethanol fermentations.
Ricardo Lagoa, Dorinda Marques-da-Silva, Mário Diniz, Maria Daglia, and Anupam Bishayee
Elsevier BV
Carolina Madeira, Marta Dias, Ana Ferreira, Raúl Gouveia, Henrique Cabral, Mário S. Diniz, and Catarina Vinagre
Frontiers Media SA
Ocean warming has been a major driver of coral reef bleaching and mass mortality. Coupled to other biotic pressures, corals’ ability for acclimatization and adaptation may become compromised. Here, we tested the combined effects of warming scenarios (26, 30, and 32°C) and predation (wound vs. no wound) in coral health condition (paleness, bleaching, and mortality), cellular stress responses (heat shock protein 70 kDa Hsp70, total ubiquitin Ub, and total antioxidant capacity TAC), and physiological state (integrated biomarker response index, IBR) of seven Scleractinian coral species, after being exposed for 60 days. Results show that although temperature was the main factor driving coral health condition, thermotolerant species (Galaxea fascicularis, Psammocora contigua, and Turbinaria reniformis) displayed increased paleness, bleaching, and mortality in predation treatments at high temperature, whereas thermosensitive species (Acropora tenuis, Echinopora lamellosa, and Montipora capricornis brown and green morphotypes) all died at 32°C, regardless of predation condition. At the molecular level, results show that there were significant main and interactive effects of species, temperature, and predation in the biomarkers assessed. Temperature affected Hsp70, Ub, and TAC, evidencing the role of protein folding and turnover, as well as reactive oxygen species scavenging in heat stress management. Predation increased Hsp70 and Ub, suggesting the activation of the pro-phenoloxidase system and cytokine activity, whereas the combination of both stressors mainly affected TAC during moderate stress and Ub under severe stress, suggesting that redox balance and defense of homeostasis are crucial in tissue repair at high temperature. IBR levels showed an increasing trend at 32°C in predated coral fragments (although non-significant). We conclude that coral responses to the combination of high temperature and predation pressure display high inter-species variability, but these stressors may pose a higher risk of endosymbiont loss, depending on species physiology and stress intensity.
Roberta G. Leitão, Maria P. Silva, Mario S. Diniz, and Mauro Guerra
Elsevier BV
Diana Madeira, Joana Filipa Fernandes, Daniel Jerónimo, Patrícia Martins, Fernando Ricardo, Andreia Santos, Maria Rosário Domingues, Mário Sousa Diniz, and Ricardo Calado
Elsevier BV
Carolina Madeira, Diana Madeira, Nemiah Ladd, Carsten J. Schubert, Mário S. Diniz, Catarina Vinagre, and Miguel C. Leal
Elsevier BV
Ana Rita Lopes, Cátia Figueiredo, Eduardo Sampaio, Mário Diniz, Rui Rosa, and Tiago F. Grilo
Elsevier BV