Conceição Santos

University of Porto, Oporto, Porto, Portugal

Are you Conceição Santos?

Claim your profile

Publications (103)226.62 Total impact

  • [Show abstract] [Hide abstract]
    ABSTRACT: Lycopene has been reported as the antioxidant most quickly depleted in skin upon UV irradiation, and thus it might play a protective role. Our goal was to investigate the effects of preexposure to lycopene on UV-B-irradiated skin cells. Cells were exposed for 24 h to 10 M lycopene, and subsequently irradiated and left to recover for another 24 h period. Thereafter, several parameters were analyzed by FCM and RT-PCR: genotoxicity/clastogenicity by assessing the cell cycle distribution; apoptosis by performing the Annexin-V assay and analyzing gene expression of apoptosis biomarkers; and oxidative stress by ROS quantification. Lycopene did not significantly affect the profile of apoptotic, necrotic and viable cells in nonirradiated cells neither showed cytostatic effects. However, irradiated cells previously treated with lycopene showed an increase in both dead and viable subpopulations compared to nonexposed irradiated cells. In irradiated cells, lycopene preexposure resulted in overexpression of BAX gene compared to nonexposed irradiated cells. This was accompanied by a cell cycle delay at S-phase transition and consequent decrease of cells in G0/G1 phase. Thus, lycopene seems to play a corrective role in irradiated cells depending on the level of photodamage. Thus, our findings may have implications for the management of skin cancer.
    No preview · Article · Jan 2016 · Oxidative medicine and cellular longevity
  • [Show abstract] [Hide abstract]
    ABSTRACT: Purpose Standard assays for phytotoxicity provide a reductionist view on the performance of plants under toxic stress. To address two of the most prominent issues in plant toxicity studies, our aims were (1) to assess how well physiological and biochemical parameters complement standard toxicological endpoints when testing natural soils and (2) to assess the suitability of three commonly used control soils as comparative references. Material and methods We compared the performance of Zea mays and Helianthus annuus in three control soils (artificial Organisation for Economic Co-operation and Development (OECD) soil, standard LUFA 2.2 soil, and turf-perlite) against three natural soils representing a gradient of contamination (from a deactivated uranium mine). Standard endpoints (emergence and biomass) were estimated, along with pigment content, photosynthetic parameters, cellular injury, and proline content. Results and discussion The toxicological profile of natural soils was highly dependent on the control soil used as reference; also, plant physiological performance was influenced by the soils’ properties. We discuss the need to interpret and combine multiple lines of evidence as a way to increase the degree of confidence one classifies soils based on their ecotoxicity, and this is where the integration of physiological and biochemical parameters bring added value. Conclusions When facing large variability in soil characteristics, it is best to collect and integrate as much information possible to strengthen conclusions about phytotoxicity of natural soils. Obviously, this refutes reductionist views and places the final conclusion in the hands of expert judgment.
    No preview · Article · Nov 2015 · Journal of Soils and Sediments
  • [Show abstract] [Hide abstract]
    ABSTRACT: Due to their antimicrobial properties, silver nanoparticles (AgNPs) are increasingly incorporated into consumer goods and medical products. Their potential toxicity to human cells is however a major concern, and there is a need for improved understanding of their effects on cell metabolism and function. Here, Nuclear Magnetic Resonance (NMR) metabolomics was used to investigate the metabolic profile of human epidermis keratinocytes (HaCaT cell line) exposed for 48h to 30 nm citrate-stabilized spherical AgNPs (10 and 40 μg/mL). Intracellular aqueous extracts, organic extracts and extracellular culture medium were analysed to provide an integrated view of the cellular metabolic response. The specific metabolite variations, highlighted through multivariate analysis and confirmed by spectral integration, suggested that HaCaT cells exposed to AgNPs displayed upregulated glutathione-based antioxidant protection, increased glutaminolysis, downregulated tricarboxylic acid (TCA) cycle activity, energy depletion and cell membrane modification. Importantly, most metabolic changes were apparent in cells exposed to a concentration of AgNPs which did not affect cell viability at significant levels, thus underlying the sensitivity of NMR metabolomics to detect early biochemical events, even in the absence of a clear cytotoxic response. It can be concluded that NMR metabolomics is an important new tool in the field of in vitro nanotoxicology.
    No preview · Article · Sep 2015 · Archives of Biochemistry and Biophysics
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Calendula L. is a circum-Mediterranean genus that comprises 10 to 25 species, depending on the taxonomic concept adopted. Several partial taxonomic revisions have been done, but none included the entire genus, and no agreement has been reached on its infrageneric classification. The inconsistent taxonomic concepts preconized by the different authors is a consequence of the large morphological variability of the genus (including the still not very well understood phenomenon of heterocarpy), of the occurrence of intermediate forms and of a large cytological variability (2n = 14 to 88 chromosomes, genome sizes and ploidy levels). To contribute to a better classification and understanding of the evolutionary relationships of the taxa of this genus, morphometric analysis, karyology and flow cytometry were used in plants extensively collected in the field, complemented with herbarium vouchers and plants obtained through seeds. The results obtained so far enabled us to delimit four species: C. arvensis, C. tripterocarpa, C. officinalis and C. suffruticosa. In the latter we included nine subspecies, including the formerly recognized species C. incana. As C. incana shares the same chromosome number and genome size as C. suffruticosa, the former species was considered a subspecies of C. suffruticosa and not a distinct species. This was further reinforced by the results of molecular analysis (O. Plume, pers. com.) which also do not support the segregation in two groups. In this work we present a summary of the data that led us to propose this classification of the genus for the Iberian Peninsula.
    Full-text · Poster · Aug 2015
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Germplasm preservation plays an important role in current breeding programs. A simple vitrification procedure that allows for the reproducible cryopreservation of two alder embryogenic lines is presented for the first time. Somatic embryos clumps (1–2 mm) were precultured in hormone-free medium (Murashige and Skoog half-strength macronutrients, MS1/2) supplemented with 0.3 M sucrose for 3 days, and then treated with a mixture of 2 M glycerol plus 0.4 M sucrose for 20 min at 25 °C. Osmoprotected somatic embryos were dehydrated using plant vitrification solution 2 (PVS2). The effect of different PVS2 incubation times was evaluated, and 60 min at 0 °C was considered to be the optimum period. After changing the solution with fresh PVS2, the somatic embryos were directly immersed in liquid nitrogen. Following rapid thawing in a water bath at 40 °C for 2 min, the somatic embryos were transferred onto MS1/2 supplemented with 0.1 mg l−1 benzyladenine, 30 g l−1 sucrose and 6 g l−1 Vitro agar. The cultures were kept in the dark for 1 week prior to exposure to light (16 h/8 h light/dark cycle). The recovery rate of vitrified somatic embryos reached over 90 % in both embryogenic lines. Cryopreservation did not affect the plant regeneration potential of Alnus glutinosa through somatic embryogenesis. The ploidy stability of the regenerated material was assessed by flow cytometry. Analysis of DNA ploidy stability of the control, PVS2 treated, cryopreserved somatic embryos and plantlets showed no significant differences.
    Full-text · Article · Aug 2015 · Plant Cell Tissue and Organ Culture
  • [Show abstract] [Hide abstract]
    ABSTRACT: Drumstick (Moringa oleifera L.) is distributed mostly in tropics/subtropics. This species have high nutritional and medicinal value. Also, interest on its industrial applications is emerging, supporting its putative economic value. Particularly, drumstick occurs in regions under desertification risk (e.g. Africa, South America) and has been for centuries a nutritional source (it provides a rich and unusual combination of important minerals, vitamins, amino acids, phenolic compounds and protein) for local populations. However, still little is known on its metabolome profile, and on the influence that UVB rays (increasing due to ozone depletion) and drought may have on drumstick. The aim of this work is to characterize metabolic changes in leaves of 2-month old drumstick exposed to drought stress (DS) and to UVB stress, (alone or combined), and subjected to a recovery period of 10 days. For that, spectrophotometric analyses and GC-MS, HPLC-MS will be used. Some of the spectrophotometric data on sugars and pigments indicate that total soluble sugars (TSS) increase with dissection (e.g. may have an osmotic role). Curiously, plants exposed to DS followed by UVB stress showed during the recovery phase an increase in TSS content, supporting that these conditions influence the metabolism of drumstick, but at different stages of exposure. Also, starch levels increased in the UVB-treated leaves, while decreasing during the recovery phase, supporting that UVB may directly stimulate metabolism associated with starch synthesis, or impair its degradation. However, the removal of UVB leads to a turnover of this metabolic response, decreasing the levels of starch. In UV stressed plants the content of pigments increases, an effect that was stabilized during the recovery. The preliminary data obtained in the present study is being confirmed by mass spectroscopy data analyses, but overall supports that both DS and UVB stress influence drumstick metabolism, being UVB more severe overall than DS. Acknowledgements: The authors thanks to FCT to support MCDias (SFRH/BPD/100865/2014) and the Ministério da Agricultura de Timor-Leste. FMachado and MAraujo contributed equally to this work.
    No preview · Conference Paper · Mar 2015
  • Márcia Araújo · Maria Costa · Conceição Santos · Maria Celeste Dias
    [Show abstract] [Hide abstract]
    ABSTRACT: Extreme abiotic factors like drought or UVB is influencing climate, and impact plant production. Moringa (Moringa oleifera L.) is a woody species (Moringaceae) native from sub-Himalaya region and currently worldwide spread particularly in tropical countries and in regions under high climate stress pressure (e.g. south Asia). The interest on this species is emerging due to its multiple ethnobotanical uses, and its nutritional value. However the impact of climate changes on this species remains unclear. In this work, we focus on how drought and UVB factors - alone or combined - can affect photosynthesis and oxidative homeostasis of M. oleifera, and the plant’s ability to recover after stress imposition. For that, 2 month old plants of M. oleifera were subjected (under greenhouse conditions) to the following treatments: drought stress (DS), UVB (UVB), DS+UVB and recovery (R, 10 days). Parameters associated with photosynthesis (F0, Fv/Fm), and with oxidative stress (malondialdehyde, MDA) and membrane damage (cell Membrane Permeability, CMP) and were determined. Preliminary data support that the UVB stress seems to have more impact than the others stresses, as all parameters were more drastically affected under this condition. Curiously, plants pretreated with DS and then exposed to UVB (DS+UVB) showed lower influence on photosynthetic parameters and on oxidative stress damages, supporting that water deficit may be used to acclimate plants to UVB stress. This hypothesis is currently under investigation (e.g. increase of the antioxidant stress). Also, our data demonstrate that studying the combination of these factors within a climate change perspective is fundamental to understand their impact on Moringa production. Acknowledge: The authors thanks to FCT to support MC Dias (SFRH/BPD/100865/2014) and Ministério da Agricultura de Timor Leste.
    No preview · Conference Paper · Mar 2015
  • Conceição Santos · Sónia Silva · Olinda Pinto-Carnide
    [Show abstract] [Hide abstract]
    ABSTRACT: The increase of aluminum (Al) bioavailability in the soil seriously compromises plant growth and crop production. Al phytotoxicity highly depends on the interactions between Al and root cells, apoplast and symplast, which are influenced by genotype and environmental conditions (e.g., soil pH, nutrients availability). Within physiological parameters used to assess Al phytotoxicity, root elongation is widely known, either by cell wall extensibility and cell cycle impairment. Besides, recent data gave new insight on the relations between Al and nutrients, Al allocation in apoplast/symplast, cell oxidative stress homeostasis, cell differentiation, and photosynthesis. We also discuss some of the recent advances on plant tolerance including genetic mechanisms for Al exclusion, where organic acids play a crucial role, and mechanisms involved in Al compartmentation and tolerance. Important advances on the interaction of Al with genes regulating these physiological events are also highlighted.
    No preview · Article · Dec 2014 · Advances in Molecular Toxicology
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Cadmium (Cd) is a widespread heavy metal used in numerous industrial processes. Cd exerts toxicological effects mostly in kidney and liver. Bone is also an important target of Cd, however, the cellular mechanisms of Cd toxicological effects in the bone cells are still poorly understood. Therefore, the present work aimed to investigate the putative cytotoxic and genotoxic effects of Cd to human bone cells. For that, the osteoblast-like MG-63 cells were exposed to 20 and 50 μM Cd for 24 and 48 h. Results showed a dose-dependent increase in Cd accumulation in cells and a decrease in cell viability, especially after 48 h. Cell cycle analysis showed a delay at S phase concomitant with a decrease in cells at G0/G1 phase. After 24 h, Cd treatment downregulated the expression of CHEK1, CHEK2 and CDK2 genes and upregulated the expression of CCNE1 gene. After 48 h, the expression of ATM and CCNB1 genes were downregulated. Also, a 3.3 fold increase on the expression of gene CCNE1 was detected. Both Cd doses induced DNA fragmentation at 48 h, while an increase in micronuclei (MN) and nucleoplasmic bridges (NPBs) together with an increase in the percentage of apoptotic/necrotic cells was detected for both time periods. Overall, our results demonstrate the cytotoxicity and genotoxicity of Cd in human bone cells. Also, the cytokinesis-block micronucleus (CBMN) assay parameters (MN, NPBs and the percentage of cells under apoptosis or necrosis) together with the cell cycle appear as the most sensitive to Cd cyto- and genotoxicity, being early affected even with the lowest Cd dose. Therefore, these cyto-/genotoxic techniques may be selected for early detection of Cd-induced toxicity.
    Full-text · Article · Oct 2014 · Mutation Research/Genetic Toxicology and Environmental Mutagenesis

  • No preview · Conference Paper · Sep 2014

  • No preview · Conference Paper · Sep 2014
  • [Show abstract] [Hide abstract]
    ABSTRACT: Unlabelled: BACKGROUND & SCOPE OF THE REVIEW: This review focuses on the UV radiation effects on skin, emphasizing the photoaging process, and the photoprotection conferred by tretinoin (all-trans retinoic acid or ATRA). Tretinoin is still the best tested retinoid to reverse photoaged skin. Major conclusions: Tretinoin can be used for photoaging treatment or combined treatment by different mechanisms. It binds to and activates retinoic acid receptors, inducing changes in gene expression that leads to cell differentiation, decreased cell proliferation, and inhibition of tumourigenesis. It has been demonstrated that photoaging resulting from UV-B radiation can be treated by retinoid formulations. Pretreatment of human skin with tretinoin blocks dermal matrix degradation followed by sun exposure, inhibiting the induction of the activated protein-1 (AP-1) transcription factor and AP-1 regulated matrix-degrading metalloproteinases. GENERAL SIGNIFICANCE AND INTEREST: Tretinoin should be considered as a key factor as it is the most potent and best-studied retinoid. In addition, the development of advanced drug delivery systems, especially novel nanoformulations, has contributed to overpass some technical drawbacks besides the skin irritation potential. The triple combination of tretinoin, hydroquinone and corticosteroids is still considered the gold standard for melasma. Although there are other novel therapeutic approaches, more high-quality clinical trials are still needed.
    No preview · Article · Aug 2014 · Mini Reviews in Medicinal Chemistry
  • Source

    Full-text · Dataset · Jul 2014
  • [Show abstract] [Hide abstract]
    ABSTRACT: Background & Scope of the reviewThis review focuses on the photoprotection conferred by lycopene, one of the most potent antioxidants. Lycopene has been recently proposed to play a critical role on anticarcinogenic action at different levels.Major Conclusions The photoprotective properties of lycopene remain contradictory. Some studies point out a positive and others a negative effect in both in vitro and in vivo models. Currently, researchers recognize that crucial gaps exist in understanding the role of carotenoids as effective modulators of apoptosis, cell cycle dynamics and/or of their in vivo behavior as cellular antioxidants.General SignificanceThe development of novel therapeutic strategies for skin disorders depends on our understanding of the molecular mechanism of UV damage on skin cells. The use of several effective phytocompounds, including lycopene, working through preventive and/ or corrective pathways in the cell, may be an approach for reducing UV-B generated damage.This article is protected by copyright. All rights reserved.
    No preview · Article · Jul 2014 · Experimental Dermatology
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Introduction Ultradeformable vesicles are highly promising tools to enhance the percutaneous transport of different drugs such as tretinoin across the skin barrier and also to increase the formulation stability at absorption site and reduce the drug induced irritation. Methods Topical delivery of tretinoin-loaded ultradeformable vesicles (tretinoin-UDV) was evaluated concerning different studies, such as: the release and permeation profiles (tape stripping); skin penetration (fluorescence analysis); induced electrical changes in skin barrier properties; cytotoxicity (Trypan Blue assay) and skin irritation in in vivo conditions (Draize test). The novel formulation performance was also compared to a commercial tretinoin formulation regarding in vivo studies. Results It was obtained a sustained and controlled drug release, as expected for UDV formulation. In addition, a dermal delivery was observed regarding the permeation study since it was not detected any drug amount in the receptor phase after 24 h. Nile Red-UDV stained intensively mostly in the stratum corneum, corroborating the tape stripping results. Tretinoin-UDV decreased skin resistance, suggesting its ability to induce skin barrier disruption. Finally, the formulation vehicle (empty UDV) and tretinoin-UDV were not toxic under in vitro and in vivo conditions, at least, at 5 × 10-3 mg/mL and 0.5 mg/mL of tretinoin, respectively. Conclusion Tretinoin-UDV is a promising delivery system for tretinoin dermal delivery without promoting skin irritation (unlike other commercial formulations), which is quite advantageous for therapeutic purpose.
    Full-text · Article · May 2014 · European Journal of Pharmaceutics and Biopharmaceutics
  • Maria Celeste Dias · Helena Oliveira · Armando Costa · Conceição Santos
    [Show abstract] [Hide abstract]
    ABSTRACT: Hormonal conditioning of plants in order to increase photosynthetic performance and reduce oxidative stress may improve plants’ tolerance to stress. This study aims to elucidate the effects of ABA pretreatment on the photosynthetic apparatus and antioxidant battery of Ulmus minor plants under well watered (WW) and drought stress (DS) conditions. Leaves were sprayed with ABA (50 and 100 μM). After 25 days of treatment DS was initiated by withholding water for 6 days. Water deficit decreased the RWC, induced stomatal closure and impaired net CO2 assimilation rate (A). However, independently of the water regime, ABA pretreatment increased plant DW accumulation, A, carotenoids and Chl a contents and reduced water loss. DS induced oxidative stress, but ABA application increased DS tolerance by the enhancement of the antioxidant system. Under WW conditions, the benefits of ABA application in reducing the cell membrane damages were noticeable. ABA pretreatment and DS induced changes in U. minor cell cycle of leaf cells, with a delay in S phase and an increase of FPCV coefficient. We propose that ABA pretreatment improves plant performance by increasing plant DW accumulation and augmenting the antioxidant system of U. minor plants, not only under DS conditions, but also under WW conditions. The use of ABA as pretreatment to alleviate the negative effects of DS seems to be a promising strategy to reduce plant's loss and improve plant productivity in drought prone habitats.
    No preview · Article · Apr 2014 · Environmental and Experimental Botany
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Sulforaphane (SFN) is a naturally-occurring isothiocyanate best known for its role as an indirect antioxidant. Notwithstanding, in different cancer cell lines, SFN may promote the accumulation of reactive oxygen species (ROS) and cause cell death e.g. by apoptosis. Osteosarcoma often becomes chemoresistant, and new molecular targets to prevent drug resistance are needed. Here, we aimed to determine the effect of SFN on ROS levels and to identify key biomarkers leading to ROS unbalance and apoptosis in the p53-null MG-63 osteosarcoma cell line. MG-63 cells were exposed to SFN for up to 48 h. At 10 μM concentration or higher, SFN decreased cell viability, increased the%early apoptotic cells and increased caspase 3 activity. At these higher doses, SFN increased ROS levels, which correlated with apoptotic endpoints and cell viability decline. In exposed cells, gene expression analysis revealed only partial induction of phase-2 detoxification genes. More importantly, SFN inhibited ROS-scavenging enzymes and impaired glutathione recycling, as evidenced by inhibition of glutathione reductase (GR) activity and combined inhibition of glutathione peroxidase (GPx) gene expression and enzyme activity. In conclusion, SFN induced oxidative stress and apoptosis via a p53-independent mechanism. GPx expression and activity were found associated with ROS accumulation in MG-63 cells and are potential biomarkers for the efficacy of ROS-inducing agents e.g. as co-adjuvant drugs in osteosarcoma.
    Full-text · Article · Mar 2014 · PLoS ONE
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Cork oak (Quercus suber L.) is a research priority in the Mediterranean area and because of cork oaks’ distribution these stands are experiencing daily stress. Based on projections of intensifying climate change and considering the key role of exploring the recovery abilities, cork oak seedlings were subjected to a cumulative temperature increase from 25 °C to 55 °C and subsequent recovery. CO2 assimilation rate, chlorophyll fluorescence, anthocyanins, proline and lipid peroxidation were used to evaluate plant performance, while the relative abundance of seven genes encoding for proteins of cork oak with a putative role in thermal/stress regulation (POX1, POX2, HSP10.4, HSP17a.22, CHS, MTL and RBC) was analyzed by qPCR (quantitative Polymerase Chain Reaction). A temperature change to 35 °C showed abundance alterations in the tested genes; at 45 °C, the molecular changes were associated with an antioxidant response, possibly modulated by anthocyanins. At 55 °C, HSP17a.22, MTL and proline accumulation were evident. After recovery, physiological balance was restored, whereas POX1, HSP10.4 and MTL abundances were suggested to be involved in increased thermotolerance. The data presented here are expected to pinpoint some pathways changes occurring during such stress and further recovery in this particular Mediterranean species.
    Full-text · Article · Mar 2014 · Journal of plant physiology
  • Source
    [Show abstract] [Hide abstract]
    ABSTRACT: Osteosarcoma is a recalcitrant bone malignancy with poor responsiveness to treatments; therefore, new chemotherapeutic compounds are needed. Sulforaphane (SFN) has been considered a promising chemotherapeutic compound for several types of tumors by inducing apoptosis and cytostasis, but its effects (e.g., genotoxicity) in osteosarcoma cells remains exploratory. In this work, the MG-63 osteosarcoma cell line was exposed to SFN up to 20 μM for 24 and 48 h. SFN induced G2/M phase arrest and decreased nuclear division index, associated with disruption of cytoskeletal organization. Noteworthy, SFN induced a transcriptome response supportive of G2/M phase arrest, namely a decrease in Chk1- and Cdc25C-encoding transcripts, and an increase in Cdk1-encoding transcripts. After 48-h exposure, SFN at a dietary concentration (5 μM) contributed to genomic instability in the MG-63 cells as confirmed by increased number of DNA breaks, clastogenicity, and nuclear and mitotic abnormalities. The increased formation of nucleoplasmic bridges, micronuclei, and apoptotic cells positively correlated with loss of viability. These results suggest that genotoxic damage is an important step for SFN-induced cytotoxicity in MG-63 cells. In conclusion, SFN shows potential to induce genotoxic damage at low concentrations and such potential deserves further investigation in other tumor cell types.
    Full-text · Article · Jan 2014 · Nutrition and Cancer
  • [Show abstract] [Hide abstract]
    ABSTRACT: This paper describes a proton high resolution magic angle spinning (HRMAS) nuclear magnetic resonance (NMR) metabolomic study of lettuce (Lactuca sativa L) leaves to characterise metabolic adaptations during leaf growth and exposure to mancozeb. Metabolite variations were identified through multivariate analysis and checked through spectral integration. Lettuce growth was accompanied by activation of energetic metabolism, preferential glucose use and changes in amino acids, phospholipids, ascorbate, nucleotides and nicotinate/nicotinamide. Phenylalanine and polyphenolic variations suggested higher oxidative stress at later growth stages. Exposure to mancozeb induced changes in amino acids, fumarate and malate, suggesting Krebs cycle up-regulation. In tandem disturbances in sugar, phospholipid, nucleotide and nicotinate/nicotinamide metabolism were noted. Additional changes in phenylalanine, dehydroascorbate, tartrate and formate were consistent with a higher demand for anti-oxidant defence mechanisms. Overall, lettuce exposure to mancozeb was shown to have a significant impact on plant metabolism, with mature leaves tending to be more extensively affected than younger leaves.
    No preview · Article · Jan 2014 · Food Chemistry

Publication Stats

1k Citations
226.62 Total Impact Points

Institutions

  • 2015
    • University of Porto
      • Faculty of Sciences
      Oporto, Porto, Portugal
  • 2014
    • Centro de Estudios y Servicios Ambientales
      Cuba
  • 2004-2014
    • University of Aveiro
      • • Division of Biology
      • • Centre for Environmental and Marine Studies (CESAM)
      Aveiro, Aveiro, Portugal