BLUE YOUNG TALENT PLUS BYTplus

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1 BLUE YOUNG TALENT PLUS BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL CIIMAR PROPOSTA DE PROJETO DE MESTRADO ORIENTADOR(A)/SUPERVISOR: Agostinho Antunes (Líder de Grupo CIIMAR/Professor FCUP) GRUPO: Genómica Evolutiva e Bioinformática LOCAL DE REALIZAÇÃO DO TRABALHO: CIIMAR/FCUP TEMA/Theme: SeXomics Sexo e o meio ambiente: Descodificação genómica e a perpetuação da vida animal num mundo em mudança / SeXomics Sex and the environment: Genomic decoding and the perpetuation of animal life in a changing world RESUMO/SUMMARY RESUMO: A maioria dos animais reproduzem se sexualmente, mas existe uma notável diversidade de mecanismos determinantes do sexo, mesmo entre espécies intimamente relacionadas. Por exemplo, em algumas espécies as mudanças de sexo podem ocorrer dentro da vida de um indivíduo e incluem hermafroditas e o sexo pode ser controlado no nível do cromossoma ou através de pistas ambientais. Estes processos têm importantes implicações evolutivas e ecológicas nos padrões de variação genética, grau de endogamia, taxa de adaptação a novos ambientes e taxas de impacto e riscos de especiação e extinção. No entanto, além de algumas espécies modelo, sabemos pouco sobre por que e como diferentes sistemas sexuais evoluíram, particularmente em vertebrados inferiores e invertebrados. Estudos comparativos têm sido dificultados pela falta de dados biológicos e genómicos acessíveis sobre sistemas sexuais, mecanismos de determinação de sexo e diversificação na árvore animal da vida e pela falta de compreensão dos processos ambientais e genómicos envolvidos na evolução dos sistemas de acasalamento. Tais dados permitirão não só analisar em profundidade a evolução dos sistemas sexuais, como também analisar mais detalhadamente o risco de mudança ambiental na distribuição, invasividade e extinção de espécies. OBJECTIVOS/OBJECTIVES: Aqui, propomos caracterizar os padrões genéticos de genes associados à reprodução em espécies de metazoárias sob diferentes pressões selectivas ambientais, e diversificação sexual associada à adaptação ambiental nos níveis de escala ampla / fina. Estas questões incluem: (1) A determinação do sexo no ambiente coloca as espécies em alto risco de extinção, especialmente num mundo em mudança do clima; (2) Alguns sistemas sexuais são mais transitórios do que outros; e (3) Como os mecanismos de determinação do sexo afectam os padrões de evolução genómica? Propomos a utilização de análises de sequenciação de última geração de subgenomes reprodutivos (gene / nível cromossómico) e 2) análises proteómicas para testar interacções proteicas funcionais e putativas com poluentes ambientais para obter dados críticos de padrões de evolução de espécies, diversificação e mal funcionamento genético associado. Os dados genómicos fornecerão valor e ferramentas importantes para pesquisas ambientais e biomédicas, possivelmente levando a novas aplicações, incluindo terapias genéticas e regenerativas mais eficientes, desenho de novas drogas, novos mecanismos de controlo de pragas, etc.

2 PRINCIPAIS METODOLOGIAS/MAIN METHODOLOGIES: Identificação de substituições sinónimas (silenciosas) e não sinónimas (substituições aminoacídicas) para testar evidências de seleção a nível molecular utilizando testes estatísticos robustos. Interações funcionais importantes em sistemas enzimáticos (complexo recetor ligando) e interações proteína proteína serão estudados em detalhe atómico. A integração final destes resultados irá permitir a simulação de vários aspetos das complexas interações biológicas que influenciam a evolução do sexo, que poderão ser validados através de testes funcionais experimentais com proteínas recombinantes. O significado evolutivo de determinadas mutações poderá ser de particular interesse para a saúde humana e investigação farmacêutica. Finalmente, os dados também serão usados para reavaliar a evolução e as relações filogenéticas entre várias espécies de metazoários, o que pode fornecer valor sobre a capacidade das espécies de sobreviver, adaptar e/ou ter sucesso em ambientes degradados e em mudança afectados por poluentes ambientais ou mudança climática. Tarefas a desempenhar pelo estudante que se integrará na equipa deste projeto de investigação (PTDC/CTA AMB/31774/2017) financiado pela FCT: Análise laboratorial e bioinformática de genes envolvidos na reprodução e sexo em metazoários (vertebrados e invertebrados). O trabalho experimental incluirá a recolha de organismos e extração de DNA/RNA, PCR, sequenciação e outras metodologias de genómica molecular. Interpretação dos resultados obtidos utilizando métodos filogenéticos e de adaptação molecular. Participação na elaboração de manuscritos. REFERÊNCIAS/REFERENCES: BORGES R, JOHNSON WE, O BRIEN SJ, VASCONCELOS V, ANTUNES A The emergence and the duplication events in the melanopsin gene family (OPN4m and OPN4x) during vertebrate evolution. PLoS One 7(12): e (Factor de impacto: 4.092). PHILIP S, MACHADO JP, MALDONADO E, VASCONCELOS V, O'BRIEN SJ, JOHNSON WE, ANTUNES A. (2012). Fish lateral line innovation: insights into the evolutionary genomic dynamics of a unique mechanosensory organ. Molecular Biology and Evolution 29(12): (Factor de impacto: ). SUNAGAR K, JOHNSON WE, O BRIEN SJ, VASCONCELOS V, ANTUNES A. (2012) Evolution of CRISPs associated with toxicoferan reptilian venom and mammalian reproduction. Molecular Biology and Evolution 29(7): (Factor de impacto: ). DUTERTRE S, JIN A, VETTER I, HAMILTON B, SUNAGAR K, LAVERGNE V, DUTERTRE V, FRY BG, ANTUNES A, VENTER D, ALEWOOD P, LEWIS R (2014) Evolution of separate predation and defence evoked venoms in carnivorous snails. Nature communications 5:3521. (Factor de impacto: ). ZHANG G, LI C, LI Q, LI B, LARKIN DM, LEE C, STORZ JF, ANTUNES A, GREENWOLD MJ, MEREDITH RW, ET AL Comparative genomics reveals insights into avian genome evolution and adaptation. Science. 346(6215): (Factor de impacto: ). KHAN I, YANG Z, MALDONADO E, LI C, ZHANG G, GILBERT MTP, JARVIS ED, O BRIEN SJ, JOHNSON WE, ANTUNES A. (2015) Olfactory receptor subgenomes linked with broad ecological adaptations in Sauropsida. Molecular Biology and Evolution 32(11): (Factor de impacto: ). BORGES R, KHAN I, JOHNSON WE, GILBERT MTP, ZHANG G, JARVIS ED, O BRIEN SJ, ANTUNES A. (2015) Gene loss, adaptive evolution and the co evolution of plumage coloration genes with opsins in birds. BMC Genomics 16(1):751. (Factor de impacto: 3.990). SILVA L, ANTUNES A. (2017) Vomeronasal Receptors in Vertebrates and the Evolution of Pheromones Detection. Annual Review of Animal Bioseciences 5: (Factor de impacto: 4.348). BORGES R, JOHNSON WE, O BRIEN SJ, HEESY C, ANTUNES A. (2018) Adaptive genomic evolution of opsins reveals that early mammals flourished in nocturnal environments. BMC Genomics 19: 121. (Impact factor: 3.867). MARRA NJ, STANHOPE MJ, JUE N, WANG M, SUN Q, BITAR PP, RICHARDS VP, KOMISSAROV A, RAYKO M, KLIVER S, STANHOPE BJ, WINKLER C, O BRIEN SJ, ANTUNES A, JORGENSEN S, SHIVJI MS (2019) White shark genome: ancient elasmobranch adaptations associated with wound healing and the maintenance of genome stability. PNAS. Outros trabalhos do grupo publicados: Referências na comunicação social: de genomas de 48 especies revela como evoluiram os passaros_n789258# do genoma do grande tubarao branco tem adn da u porto/ tubaraobranco deixounos longevidade #gs.i3XgfDqg ÁREA CIENTÍFICA: Genómica, Genética, Evolução Molecular, Bioinformática. LICENCIATURAS ADMITIDAS: Biologia, Bioquímica, Ciências dos Computadores. Outras licenciaturas afins na área poderão eventualmente ser consideradas.

3 BLUE YOUNG TALENT PLUS - BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL - CIIMAR PROPOSTA DE PROJETO DE MESTRADO ORIENTADOR(A)/SUPERVISOR: Filipe Fernandes Coutinho GRUPO/GROUP: NUTRIMU - CIIMAR CO-ORIENTADOR(A)/CO-SUPERVISOR: Cláudia R. Serra GRUPO/GROUP: NUTRIMU - CIIMAR LOCAL DE REALIZAÇÃO DO TRABALHO/PLACE OF WORK: CIIMAR and FCUP TEMA/THEME: Cyanobacterial outer membrane vesicles as a novel technology for European sea bass juveniles vaccination against Mycobacterium marinum RESUMO/SUMMARY This work aims at implementing outer membrane vesicles (OMV) as novel platforms in vaccine technology. The goal will be to explore OMV derived from non-pathogenic bacteria (cyanobacteria) as an antigen delivery vehicle to immunize the European seabass against the opportunistic pathogen Mycobacterium marinum that causes mycobacteriosis, for which no vaccine or satisfactory treatment is available. This work will be dedicated to test the immunostimulatory capacity of OMVs loaded with Mycobacterium marinum-proteins. It will be divided in four tasks: RESUMO: Task 1 - Fish husbandry Optimal husbandry conditions following European sea bass requirements will be applied for cultivation at CIIMAR facilities. ESB juveniles (5-10 g) will be obtained from commercial farms and after a quarantine period, will be maintained at CIIMAR facilities in a thermoregulated (22-25ºC) marine (35 salinity) water recirculation system, on a 12h light/12h dark photoperiod. Task 2 - Fish immunization Aiming to induce specific and prolonged adaptive immune responses in European sea bass, 5 distinct combinations of OMV populations, each packaged with a specific M. marinum-proteins, will be administered to triplicate groups of fish. Juveniles European sea bass will be immunized intraperitoneally with 100 μl of OMV with the following characteristics: i) OMV carrying secreted-type antigens; ii) OMV carrying mycomembrane-associated antigens; iii) OMV carrying both secreted-type and mycomembrane associated antigens; iv) empty OMV; or v) no OMV, injected with the same volume of a sterile phosphatebuffered saline solution as the control group. Prior to vaccination, fish will be anesthetized with a 0.3 mg/l of ethylene glycol monophenyl ether. On day 0 and once a week thereafter (at days 7, 14, 21, 28 - until day 70 post-vaccination) 6 fish from each experimental group will be anesthetized for blood samples collection. At the end of the trial head-kidney from three fish per tank will also be sampled and stored in RNAlater for further analysis. Task 3 CyannoVaccine-specific immunoglobulins Blood samples will be used to determine the serum concentration of specific antibodies promoted by the vaccine, by measuring the presence of specific immunoglobulins against M. marinum antigenic proteins. For such analysis, an indirect ELISA method will be used as described in Gao et al., Task 3 Immune response The expression of several immune-related genes (pro-inflammatory cytokines, IL-1β, TNF-α and IL-6) in head-kidney will be analysed by real-time PCR to evaluate fish immune response (Azeredo et al., 2017). By the end of these tasks the OMV combination which induces the most effective adaptive immune response will be identified.

4 OBJECTIVOS/OBJECTIVES: Select the best CyanoVaccine candidate, through the identification of the OMV combination that induces the most effective adaptive immune response. PRINCIPAIS METODOLOGIAS/MAIN METHODOLOGIES: Husbandry of European sea bass juveniles Immunization of European sea bass juveniles by intraperitoneal injection of engineered outer membrane vesicles (OMV) Measurement of serum specific immunoglobulins against M. marinum antigenic proteins by indirect ELISA Determination of the expression of several immune-related genes in head-kidney by real-time PCR REFERÊNCIAS/REFERENCES: Azeredo R, Pérez-Sánchez J, Sitjà- Bobadilla A, Fouz B, Tort L, Aragão C, et al., European sea bass (Dicentrarchus labrax) immune status and disease resistance are impaired by arginine dietary supplementation. PLoS ONE 10(10): e Gao BL, Liu J, Dong LX, Zhang L, Qin JH, Wang JP, Broad specific enzyme-linked immunosorbent assay for determination of residual phenothiazine drugs in swine tissues. Anal Biochem. 454:7 13. Brudal E, Lampe EO, Reubsaet L, Roos N, Hegna IK, Thrane IM, Koppang EO, Winther-Larsen HC, Vaccination with outer membrane vesicles from Francisella noatunensis reduces development of francisellosis in a zebrafish model. Fish & Shellfish Immunology 42: ÁREA CIENTÍFICA/SCIENTIFIC AREA: Biological Sciences

5 BLUE YOUNG TALENT PLUS BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL CIIMAR PROPOSTA DE PROJETO DE MESTRADO ORIENTADOR(A)/SUPERVISOR: Emília Sousa GRUPO/GROUP: Natural Products and Medicinal Chemistry CO ORIENTADOR(A)/CO SUPERVISOR: Diana Resende GRUPO/GROUP: Natural Products and Medicinal Chemistry LOCAL DE REALIZAÇÃO DO TRABALHO/PLACE OF WORK: Faculdade de Farmácia da Universidade do Porto TEMA/THEME: Exploring the sea for new antimalarial agents RESUMO/SUMMARY RESUMO: Malaria is still among the main public health problems in least developed regions of the world and, despite severe efforts amongst the scientific community in order to control and eradicate this disease, new antimalarial compounds will not become available in the upcoming years. [1,2] New evidences of a multidrug-resistant P. falciparum parasite lineage in some countries in Southeast Asia represents a serious threat to global malaria control and eradication. [1] Recently our group synthesized a series of alkaloids based on the structure of known antimalarial drugs or lead drug candidates. Some of the compounds exhibited excellent antimalarial activity against the sensitive strain P. falciparum 3D7 (IC 50 = µm), thus representing potential new chemotypes for further optimization towards novel and affordable antimalarial drugs. OBJECTIVOS/OBJECTIVES: The main purpose of this work is to obtain analogues of the active alkaloids by structure based design performing molecular modifications to obtain more efficient analogues and screening for their antimalarial activities.

6 PRINCIPAIS METODOLOGIAS/MAIN METHODOLOGIES: Organic synthesis, purification and isolation procedures and structural elucidation of organic substances. The structure elucidation will be performed by spectroscopic methods (UV, IR, 1 H and 13 C NMR), and mass spectrometry. In collaboration with other research groups, the synthesized compounds will be tested for their antimalarial activity. Analogues of the most promising compounds with potential as drug candidates or industrial applications will be prepared by molecular modifications in order to perform structure activity relationship studies. REFERÊNCIAS/REFERENCES: [1] World health organization World malaria report Available online: malaria report 2018/en/ [2] Bule MH, Ahmed I, Maqbool F, Zia MA. International Journal of Pharmacology 2017, 13(7), ÁREA CIENTÍFICA/SCIENTIFIC AREA: Medicinal Chemistry

7 BLUE YOUNG TALENT PLUS BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL CIIMAR PROPOSTA DE PROJETO DE MESTRADO ORIENTADOR(A)/SUPERVISOR: Ana Paula Mucha GRUPO/GROUP: ECOBIOTEC CO ORIENTADOR(A)/CO SUPERVISOR: Marisa Almeida GRUPO/GROUP: ECOBIOTEC LOCAL DE REALIZAÇÃO DO TRABALHO/PLACE OF WORK: CIIMAR TEMA/THEME: Assessment of microbial communities associated with lost fishing gears RESUMO/SUMMARY RESUMO: Fishing related debris represents more than 50% of the weight of the floating macroplastics pollution found in marine environment. Plastic debris provides a substrate for microbes that lasts much longer than most natural floating substrates and has been implicated as a transportation vector for biological pollutants such as opportunistic pathogens. Nevertheless, biological pollution loads in lost fishing gear (mainly composed of plastics) have not yet been sufficiently explored, being this the aim of this work. Obtained results will contribute for a better understanding of the ecological implications of fishing related debris. An important contribution will be made for the understanding of the role of lost fishing gears as a substrate for microbial colonization, with special emphasis on potential opportunistic pathogens and pollutant degrading bacteria. OBJECTIVOS/OBJECTIVES: To assess microbial communities associated with lost fishing gear, with special emphasis on 1) the presence of harmful microorganisms such as opportunistic pathogens, and 2) the identification of pollutant degrading bacteria, which can indicate the possibility of microorganism playing a role in reducing lost fishing gears impacts.

8 PRINCIPAIS METODOLOGIAS/MAIN METHODOLOGIES: Laboratory manipulative experiments will be carried out using fragments of fishing gear exposed to natural sea water under different dynamic conditions (different levels of agitation), both in the presence and in the absence of marine sediment (sand with different grain size), to simulate microbial colonization that the gear can undergo in water column and after their seafloor deposition. The microbial communities extracted from fragments of lost fishing gear will be identified by using next generation sequencing technology (NGS). Special attention will be dedicated to the presence of specific microbial groups. One of these groups are the opportunistic pathogens, as plastic debris have been implicated as vector for transportation of harmful microorganisms. The other group are pollutant degrading bacteria, or groups of microorganism presenting degradative genes, that can point to a possible role of microorganisms in reducing lost fishing gears impacts. REFERÊNCIAS/REFERENCES: Zettler, E. R., Mincer, T. J., & Amaral Zettler, L. A. (2013). Life in the plastisphere : microbial communities on plastic marine debris. Environmental science & technology, 47(13), Almeida, C. M. R., Oliveira, T., Reis, I., Gomes, C. R., & Mucha, A. P. (2017). Bacterial community dynamic associated with autochthonous bioaugmentation for enhanced Cu phytoremediation of salt marsh sediments. Marine environmental research, 132, ÁREA CIENTÍFICA/SCIENTIFIC AREA: Environmental Sciences

9 BLUE YOUNG TALENT PLUS BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL CIIMAR PROPOSTA DE PROJETO DE MESTRADO ORIENTADOR(A)/SUPERVISOR: Carla Sofia Garcia Fernandes GRUPO/GROUP: NATURAL PRODUCTS AND MEDICINAL CHEMISTRY CO ORIENTADOR(A)/CO SUPERVISOR: Maria Elizabeth Tiritan GRUPO/GROUP: NATURAL PRODUCTS AND MEDICINAL CHEMISTRY LOCAL DE REALIZAÇÃO DO TRABALHO/PLACE OF WORK: FFUP (Rua de Jorge Viterbo Ferreira n.º 228, PORTO) TEMA/THEME: Síntese de Péptidos de Origem Marinha e Análogos com Potencial Atividade Antimicrobiana/ Synthesis of Marine derived Peptides and Analogues with Potential Antimicrobial Activity RESUMO/SUMMARY RESUMO: Os oceanos são uma fonte rica de compostos bioativos estruturalmente únicos sob a perspetiva de potenciais agentes terapêuticos. Nos últimos anos, os produtos naturais marinhos têm despertado grande interesse, essencialmente devido ao leque alargado de atividades biológicas e farmacológicas que demonstram exibir. Para além disso, são interessantes modelos para modificações moleculares e / ou síntese total, de modo a obter compostos mais potentes e em maior quantidade. Os péptidos marinhos constituem um grupo particularmente interessante devido à sua química e diversas atividades biológicas, incluindo antimicrobiana. Neste contexto, esta proposta tem como objetivo sintetizar péptidos marinhos e análogos para posterior avaliação de atividade antimicrobiana. A pureza enantiomérica dos péptidos obtidos será avaliada por cromatografia líquida utilizando fases estacionárias quirais. The oceans are a rich source of structurally unique bioactive compounds under the prospect of potential therapeutic agents. In recent years, natural marine products have aroused great interest, mainly due to their wide range of biological and pharmacological activities. Additionally, they are interesting models for molecular modifications and / or total synthesis to obtain more potent compounds and in higher quantity. Marine peptides are a particularly interesting group because of their chemistry and various biological activities, including antimicrobial. In this context, this proposal aims to synthesize marine derived peptides and analogues for further evaluation of antimicrobial activity. The enantiomeric purity of the obtained peptides will be evaluated by liquid chromatography using chiral stationary phases.

10 OBJECTIVOS/OBJECTIVES: 1. Sintetizar uma pequena biblioteca de péptidos e análogos para posterior avaliação da atividade antimicrobiana 2. Elucidar a estrutura dos compostos sintetizados 3. Determinar a pureza enantiomérica dos péptidos obtidos 1. Synthesize a small library of marine derived peptides and analogues for further evaluation of antimicrobial activity 2. Structure elucidation of the synthesized compounds 3. Determination of the enantiomeric purity of the obtained peptides PRINCIPAIS METODOLOGIAS/MAIN METHODOLOGIES: Síntese orgânica de péptidos Elucidação estrutural por 1 H RMN, 13 C RMN, IV, MS e cristalografia de raios X Avaliação da pureza enantiomérica por cromatografia líquida Organic Synthesis of Peptides Structural elution by 1 H NMR, 13 C NMR, IR, MS and X ray crystallography Evaluation of the enantiomeric purity by liquid chromatography REFERÊNCIAS/REFERENCES: Phyo, Y.; Ribeiro, J.; Fernandes, C.; Kijjoa, A.; Pinto, M.M.M. Marine Natural Peptides: Determination of Absolute Configuration using Liquid Chromatography Methods and Bioactivities, Molecules, 2018, 23(2), 306, doi: /molecules Cheung, R.C.; Ng, T.B.; Wong, J.H. Marine peptides: Bioactivities and applications. Mar. Drugs 2015, 13, Gogineni, V.; Hamann, M.T. Marine natural product peptides with therapeutic potential: Chemistry, biosynthesis, and pharmacology. Biochim. Biophys. Acta, 2018, 1862, Anjum, K.; Abbas, S.Q.; Akhter, N.; Shagufta, B.I.; Shah, S.A.A.; Hassan, S.S.U. Emerging biopharmaceuticals from bioactive peptides derived from marine organisms. Chem. Biol. Drug Des., 2017, 90, ÁREA CIENTÍFICA/SCIENTIFIC AREA: Química Medicinal/Medicinal Chemistry

11 BLUE YOUNG TALENT PLUS - BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL - CIIMAR PROPOSTA DE PROJETO DE MESTRADO ORIENTADOR(A)/SUPERVISOR: Catarina Magalhães GRUPO/GROUP: EcoBioTec CO-ORIENTADOR(A)/CO-SUPERVISOR: Pedro Duarte GRUPO/GROUP: Norwegian Polar Institute CO-ORIENTADOR(A)/CO-SUPERVISOR: Maria Paola Tomasino GRUPO/GROUP: EcoBioTec LOCAL DE REALIZAÇÃO DO TRABALHO/PLACE OF WORK: CIIMAR, EcoBioTec TEMA/THEME: Biogeography of Arctic Eukaryotic Microbiome: a combining approach of metabarcoding, metagenomics and microscopic cell counts. RESUMO/SUMMARY RESUMO: Global worming and climate change has been manifested in the decrease of the Arctic Ocean see ice extent and thickness (Parkinson and Comiso 2013). The thinner sea-ice regime that the Arctic has been facing changed Arctic s primary productivity and biogeochemistry (Barber et al. 2015). In fact the dramatically sea-ice retreat in the Arctic Ocean promoted an increase in productivity, and massive under-ice phytoplankton blooms (Assmy et al. 2017). Thus a remaining question is whether those communities are likely to change in the Artic and which feedbacks might be expected from this changing eukaryotic microbiome that could accelerate or mitigate climate change impacts. OBJECTIVOS/OBJECTIVES: Understand the dynamics of the microbial communities responsible for the productivity of the Arctic ecosystem will be worthwhile in order to draw future trends on an Arctic Climate Change Scenario. This master project will aim to present a comprehensive analysis of the biogeographic patters of Arctic phytoplankton diversity and distribution along two oceanographic transects in the Marginal Ice Zone around Svalbard. By combining 5 years of unique Arctic data sets (metabarcoding/metagenomic/microscope identification) from previous ( ) and future ( ) Arctic expeditions, this study will fill critical gaps concerning the response of the main Arctic primary producers, in terms of diversity and distribution dynamics, to Arctic climate driven changes.

12 PRINCIPAIS METODOLOGIAS/MAIN METHODOLOGIES: Fieldwork will be performed in the future campaigns ( ) at the Marginal Ice Zone and Kongsfjorden integrated into a long term international monitoring program lead by Norwegean Polar Institute for Climate Change Studies in the Arctic. Phytoplankton samples will be concentrated through filtration and preserved (-80ºC) on board to be used for later DNA extraction, and genomic and metagenomic analysis using specific methodologies (Sousa et al. 2019). Microscope phytoplankton counts and identification will be performed on board in fresh samples and also in the lab in preserved samples. Sequence analyses will be performed by using illumina technology and Bioinformatic tools will be apply to analyze all the data generated from next generation sequencing (metabarcoding and metagenomics) (Sousa et al. 2019). Statistical tools will be also use to integrate phytoplankton diversity and distribution in time and space with contextualized environmental data. REFERÊNCIAS/REFERENCES: Parkinson CL, Comiso JC On the 2012 record low Arctic sea ice cover: combined impact of preconditioning and an August storm. Geophys Res Lett 40: Barber et al Selected physical, biological and biogeochemical implications of a rapidly changing Arctic Marginal Ice Zone. Progr Oceanogr 139: Assmy P et al Leads in Arctic pack ice enable early phytoplankton blooms below snow-covered sea ice. Sci. Rep. 7:40850 ÁREA Sousa CIENTÍFICA/SCIENTIFIC AG et al Diversity AREA: and Composition of Pelagic Prokaryotic and Protist Communities in a Thin Arctic Sea- Ice Regime. Microb Ecol. DOI: /s

13 BLUE YOUNG TALENT PLUS - BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL - CIIMAR PROPOSTA DE PROJETO DE MESTRADO ORIENTADOR(A)/SUPERVISOR: Maria de Fátima Carvalho GRUPO/GROUP: EcoBioTec CO-ORIENTADOR(A)/CO-SUPERVISOR: Pedro Leão; Ralph Urbatzka GRUPO/GROUP: Cyanobacterial Natural Products; Blue biotechnology and ecotoxicology LOCAL DE REALIZAÇÃO DO TRABALHO/PLACE OF WORK: EcoBioTec TEMA/THEME: Deep-sea pharmacies: Exploring deep-sea Actinobacteria for the production of novel natural products with pharmaceutical applications RESUMO/SUMMARY RESUMO: Actinobacteria are a large group of Gram-positive bacteria, highly prolific in the production of bioactive secondary metabolites with a wide range of biological and pharmaceutical properties. Most of known actinobacterial species are of terrestrial origin, but it has been recently shown that they are also true habitants of the oceans and are a proven source of novel relevant secondary metabolites, such as antibiotics, antitumor, anti-inflammatory and antiviral compounds, biosurfactants, etc. (Dharmaraj, 2010; Olano et al., 2009). The deep sea is a unique environment characterized by extreme conditions, such as high pressure, low temperature, lack of light and variable salinity and oxygen concentrations, causing unique evolutionary pressures on microorganisms. Deep-sea ecosystems are known to contain a high diversity of actinobacteria, a great proportion of which are predicted to be novel species and genera (Bull and Stach, 2007). This proposal intends to investigate the cultivable actinobacterial community associated with deep-sea samples collected in Madeira and Azores archipelagos and explore their potential to produce bioactive compounds with pharmaceutical relevance. OBJECTIVOS/OBJECTIVES: In this proposal it is intended to: (i) isolate actinobacteria from various deep-sea samples collected in Madeira and Azores islands; (ii) identify the isolated strains; (iii) investigate the potential of the isolated actinobacteria to produce compounds with antimicrobial, anticancer, anti-inflammatory and antiobesity activities; (iv) identify secondary metabolites produced by the strains exhibiting bioactivity.

14 PRINCIPAIS METODOLOGIAS/MAIN METHODOLOGIES: Several deep-sea samples recently collected in expeditions in the Madeira and Azores archipelagos are currently available in the laboratory for the isolation of actinobacteria. The cultivable community of actinobacteria associated with these samples will be isolated in different selective media and identified through 16S rrna gene sequencing. A preliminary assessment of the production of bioactive compounds by the isolates will be performed by culturing each isolate in selective liquid medium, followed by extraction of culture medium with an organic solvent and screening for antimicrobial, anticancer, anti-inflammatory and antiobesity activities in the obtained extracts. The most promising bioactive extracts will be further studied in order to identify the compound(s) responsible for the bioactivity and their chemical structures. REFERÊNCIAS/REFERENCES: - Bull, A. T. and Stach, J.E. (2007). Marine actinobacteria: new opportunities for natural product search and discovery. Trends in microbiology, 15(11): Dharmaraj, S. (2010). Marine Streptomyces as a novel source of bioactive substances. World Journal of Microbiology and Biotechnology, 26 (12): Olano, C., Méndez, C. and Salas, J.A. (2009). Antitumor compounds from marine actinomycetes. Marine Drugs, 7(2): ÁREA CIENTÍFICA/SCIENTIFIC AREA: Ciências Biológicas e afins

15 BLUE YOUNG TALENT PLUS BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL CIIMAR PROPOSTA DE PROJETO DE MESTRADO ORIENTADOR(A)/SUPERVISOR: Filipe Castro GRUPO/GROUP: AGE CO ORIENTADOR(A)/CO SUPERVISOR: Raquel Ruivo GRUPO/GROUP: LOCAL DE REALIZAÇÃO DO TRABALHO/PLACE OF WORK: CIIMAR and FCUP TEMA/THEME: When less is more: the role of gene loss in phenotypic diversification RESUMO/SUMMARY RESUMO: Genomes are dynamic biological units, with processes of gene duplication and loss triggering evolutionary novelty. Mammalian radiation entailed the successful colonization of multiple and ecologically diverse habitats. This evolutionary path was accompanied by the appearance of novel phenotypic traits in association with genomic changes such as gene duplication. Yet, Gene Inactivation has emerged as well known driver of evolutionary change. The power of this evolutionary mechanism has been described in diverse vertebrate and non vertebrate lineages. Here, we will investigate the gene composition of multiple genetic pathways (e.g. lipid and sugar metabolism) in mammals. We will combine comparative genomics with functional assays to determine when and how have key adaptations emerged in vertebrate ancestry and whether gene loss has been an effective adaptive process. OBJECTIVOS/OBJECTIVES: To investigate and identify evolutionary processes of gene loss in Vertebrate genomes and their role in phenotypic diversification. PRINCIPAIS METODOLOGIAS/MAIN METHODOLOGIES: The work plan will be entail a strong component of Bioinformatics (e.g. gene mining, gene annotation, phylogenetics, selection) in combination with Molecular and Cell Biology techniques (e.g. next generation sequencing technologies).

16 REFERÊNCIAS/REFERENCES: Marlétaz F, Firbas PN, Maeso I, Tena JJ, Bogdanovic O, Perry M, Wyatt CDR, de la Calle Mustienes E, Bertrand S, Burguera D, Acemel RD, van Heeringen SJ, Naranjo S, Herrera Ubeda C, Skvortsova K, Jimenez Gancedo S, Aldea D, Marquez Y, Buono L, Kozmikova I, Permanyer J, Louis A, Albuixech Crespo B, Le Petillon Y, Leon A, Subirana L, Balwierz PJ, Duckett PE, Farahani E, Aury JM, Mangenot S, Wincker P, Albalat R, Benito Gutiérrez È, Cañestro C, Castro F, D'Aniello S, Ferrier DEK, Huang S, Laudet V, Marais GAB, Pontarotti P, Schubert M, Seitz H, Somorjai I, Takahashi T, Mirabeau O, Xu A, Yu JK, Carninci P, Martinez Morales JR, Crollius HR, Kozmik Z, Weirauch MT, Garcia Fernàndez J, Lister R, Lenhard B, Holland PWH, Escriva H, Gómez Skarmeta JL, Irimia M. Amphioxus functional genomics and the origins of vertebrate gene regulation. Nature Dec;564(7734): doi: /s Genes for de novo biosynthesis of omega 3 polyunsaturated fatty acids are widespread in animals. Kabeya N, Fonseca MM, Ferrier DEK, Navarro JC, Bay LK, Francis DS, Tocher DR, Castro LFC, Monroig Ó. Sci Adv May 2;4(5):eaar6849. doi: /sciadv.aar6849. ÁREA CIENTÍFICA/SCIENTIFIC AREA: Comparative Genomics and Evolution LICENCIATURAS ADMITIDAS /ADMITABLE DEGREES: Biology and Biochemistry

17 BLUE YOUNG TALENT PLUS BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL CIIMAR ORIENTADOR(A)/SUPERVISOR: Carolina Castro GRUPO/GROUP: NUTRIMU CO ORIENTADOR(A)/CO SUPERVISOR: Helena Peres GRUPO/GROUP: NUTRIMU PROPOSTA DE PROJETO DE MESTRADO LOCAL DE REALIZAÇÃO DO TRABALHO/PLACE OF WORK: FCUP and CIIMAR TEMA/THEME: Potential of solid state fermented wineries and olive oil by products as functional ingredients for European sea bass feeds RESUMO/SUMMARY RESUMO: Wineries and olive oil industries are important agro industrial activities in Portugal that produce large amounts and varieties of by products (such as grape marc (GM) and olive pomace (OP)). Reuse and valorization of these agro industrial by products are crucial both from economic and environmental perspectives. Solid state fermentation(ssf) using fungi is a low cost biotechnology that allows to convert these inexpensive agro industrial by products into add value and innovative ingredients. As a source of important bioactive molecules (such as antioxidant and phenolic compounds) that are reported to induce beneficial effects on animal health and wellbeing (Fontana et al., 2013; Ghanbari et al., 2012), these ingredients have the potential to be used as functional ingredient for fish feeds. However, the potential of solid state fermented ingredients from winery and olive oil byproducts as functional ingredient in fish feeds was up to now little explored. OBJECTIVOS/OBJECTIVES: The aim of this study is to access the potential of the solid state fermentation extract of GM and OP mixture as functional ingredient in feeds for European sea bass (Dicentrarchus labrax), an important European aquaculture species. The prospective beneficial effects of the solid state fermentation extract of GM and OP mixture will be evaluated in E. sea bass in terms of zootechnical performance, physiological, immunological and oxidative status. PRINCIPAIS METODOLOGIAS/MAIN METHODOLOGIES: The first stage of this project will comprise characterization of extract obtained by solid state fermentation of the GM and OP mixture, namely the antioxidant activity, type and level of phenolic compounds present on the final product (methodology described in Dulf et al., 2015).

18 The second stage of this project will involve a growth trial to test the efficacy of the solid state fermentation extract of GM and OP mixture on fish growth, voluntary feed intake, feed efficiency and health status. A control diet will be formulated without containing the fermented extract of GM and OP mixture. Three other diets will be formulated similarly to the control diet but including increasing levels of the extract. The extract levels to be included will be determinated according to antioxidant activity, type and levels of phenolic compounds present in the extract. Each diet will be tested in triplicated. The growth trial will be run at the Marine Zoology Station in a recirculation water system, following the experimental design described in Magalhães et al. (2016). At the end of the trial blood and intestine from 3 fish per tank will be sampled to characterize: Hematological profile by assessement of erythrocyte (haematocrit; haemoglobin concentration; erythrocytes number; mean cell volume; mean cell haemoglobin; mean haemoglobin concentration) and leukocyte (total and differential leukocyte numbers) parameters as described in Machado et al. (2019). immune response by quantifying nitric oxide, total Ig and peroxidase, lysozyme and alternative complement pathway activities in plasma, according to Machado et al. (2019); and expression of genes coding for key markers of mucosal immunity (IgT), Tcell markers (CD4; CD8), pro inflammatory (IL 1β; IL 6; TNF α) and anti inflammatory (IL 10;COX 2;TGF β) cytokines in intestine by RT PCR (Machado et al., 2019); intestine oxidative status by measuring activity of antioxidant enzymes (superoxide dismutase, SOD; catalase, CAT; glutathione peroxidase, GPX; glutathione reductase, GR) and non enzymatic oxidative stress indicators (lipidperoxidation, TBARS; reduced and oxidized glutathione, GSH; GSSG), as described by Castro et al. (2015). Data on zootechnical parameters, such as growth rate, weight gain, feed intake, protein efficiency and body condition indices will be obtained. Nitrogen and energy budgets will be obtained at the end of the growth trial. REFERÊNCIAS/REFERENCES: Castro, C., Peréz Jiménez, A., Coutinho, F., Díaz Rosales, P., Serra, C.A., Panserat, S., Corraze, G., Peres, H., Oliva Teles, A Dietary carbohydrate and lipid sources affect differently the oxidative status of European sea bass (Dicentrarchus labrax) juveniles. Br. J. Nutr. 114: Dulf, F.V., Vodnar, D.C., Dulf, E., Tosa, M.I Total Phenolic Contents, Antioxidant Activities, and Lipid Fractions from Berry Pomaces Obtained by Solid State Fermentation of Two Sambucus Species with Aspergillus niger. J. Agric. Food Chem. 63: Fontana, A. R., Antoniolli, A., Bottini, R Grape pomace as a sustainable source of bioactive compounds: extraction, characterization, and biotechnological applications of phenolics. J. Agric. Food Chem. 61: Ghanbari R.,,Anwar, F., Alkharfy, K.M., Gilani, A.H., Saari, N Valuable nutrients and functional bioactives in different parts of olive (Olea europaea L.) a review. Int. J. Mol. Sci. 13: Machado, M., Castro, C., Oliva Teles, A., Costas, B Interactive effects of dietary vegetable oil and carbohydrate incorporation on the innate immune response of European seabass (Dicentrarchus labrax) juveniles subjected to acute stress. Aquaculture 498: Magalhães, R., Lopes, T., Martins, N., Díaz Rosales, P., Couto, A., Pousão Ferreira, P., Oliva Teles, A., Peres, H Carbohydrases supplementation increased nutrient utilization in white seabream (Diplodus sargus) juveniles fed high soybean meal diets. Aquaculture. 463: ÁREA CIENTÍFICA/SCIENTIFIC AREA: Aquaculture; Fish nutrition

19 BLUE YOUNG TALENT PLUS - BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL - CIIMAR PROPOSTA DE PROJETO DE MESTRADO ORIENTADOR(A)/SUPERVISOR: Inês Guerreiro GRUPO/GROUP: NUTRIMU CO-ORIENTADOR(A)/CO-SUPERVISOR: Aires Oliva-Teles GRUPO/GROUP: NUTRIMU LOCAL DE REALIZAÇÃO DO TRABALHO/PLACE OF WORK: CIIMAR and FCUP TEMA/THEME: Regulation of appetite by glucose in gilthead seabream (Sparus aurata). RESUMO/SUMMARY RESUMO: Control of feed intake and energy metabolism is crucial for proper growth and development. However, in fish, understanding on the endocrine control of appetite and body weight is not as developed as in mammals (1, 2). In aquaculture such knowledge is of vital importance, to maximize animal growth while optimizing feed conversion. Appetite regulation and metabolic utilization of diets may be affected by glucose availability. Moreover, it is known that glucose has a central role in energy homeostasis and that glucose plasmatic levels are regulated by endocrine mediators, gut-derived factors, and nutritional factors (3). Ghrelin and leptin are key hormones involved in appetite regulation; while ghrelin is involved in orexigenic responses, leptin is involved in anorexigenic responses (1). Thus, understanding how glucose interact with appetite regulating hormones, namely ghrelin and leptin, is of particular relevance when considering new aquafeeds that include high levels of plant feedstuffs rich in carbohydrates. Gilthead seabream was the chosen species since it is the most produced fish in the Mediterranean and despite its economic importance, knowledge on appetite regulation is still limited. OBJECTIVOS/OBJECTIVES: This work aims to assess glucose effect on appetite regulating hormones in gilthead seabream.

20 PRINCIPAIS METODOLOGIAS/MAIN METHODOLOGIES: Gilthead seabream will be acclimatized to the experimental conditions for 2 weeks in CIIMAR. During this time fish will be fed a commercial diet, 2 times a day, until visual satiation. Then, after 1 day of feed deprivation, fish will be intraperitoneal injected with glucose or saline solution and after 3h (corresponding to glucose peak) and 24h (post-prandial basal level) will be sampled, to assess endocrine factors involved in appetite. Fish will be sampled for blood, brain, liver, stomach, intestine and adipose tissue. The following parameters will be determined: plasma metabolites indicators of nutrient metabolism; liver composition; expression of genes related with glucose metabolism, orexigenic (orexins; NPY; ghrelin) and anorexigenic molecules (CRH; CART; CCK; leptin). This work will result in one manuscript. REFERÊNCIAS/REFERENCES: (1) Hoskins, L.J., & Volkoff, H. (2012). Gen Comp Endocrinol 176, (2) Rønnestad, I. et al. (2017). Front Endocrinol 8:73, doi: /fendo (3) Riley Jr., L.G. et al. (2009). Comp Biochem Physiol A 154, ÁREA CIENTÍFICA/SCIENTIFIC AREA: Aquaculture Fish nutrition and physiology

21 BLUE YOUNG TALENT PLUS BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL CIIMAR PROPOSTA DE PROJETO DE MESTRADO ORIENTADOR(A)/SUPERVISOR: Benjamin Costas GRUPO/GROUP: NUTRIMU CO ORIENTADOR(A)/CO SUPERVISOR: Rita Azeredo GRUPO/GROUP: NUTRIMU LOCAL DE REALIZAÇÃO DO TRABALHO/PLACE OF WORK: CIIMAR TEMA/THEME: The potential use of tryptophan supplementation for animal health management RESUMO/SUMMARY RESUMO: The concept of maintaining animal health through the best possible nutrition is well accepted in modern animal farming and functional amino acids appear to be good candidates to improve health and survival. Tryptophan in particular have known roles in the improvement of the immune response to infection and recent evidence indicates that several immune mechanisms are influenced by its availability. In this context, this project mainly aims to provide a better understanding of the underpinning neuro endocrine and immune machinery of fish fed tryptophan supplements during inflammatory conditions and to deliver a deeper understanding of the basic mechanisms involved during acute and chronic inflammatory processes. This is particularly important for the aquaculture industry, where few therapeutic possibilities are available against infectious episodes. OBJECTIVOS/OBJECTIVES: The main goal of this proposal is to allow a better understanding of the role of tryptophan on the inflammatory response, immune modulation and disease resistance in fish. In particular, it is intended to verify the neuroendocrine and immune modulatory effects of tryptophan during inflammation, and to what extent tryptophan have a significant impact on disease resistance in fish. Understanding the role of this particular amino acid during inflammation could be of major importance to the nutritional immunology community and for the design of dietary therapeutic interventions.

22 PRINCIPAIS METODOLOGIAS/MAIN METHODOLOGIES: Fish trials and fish handling procedures Hematology Assessment of immune and oxidative stress parameters Gene expression REFERÊNCIAS/REFERENCES: N/A ÁREA CIENTÍFICA/SCIENTIFIC AREA: Biological and Veterinary Sciences

23 BLUE YOUNG TALENT PLUS BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL CIIMAR PROPOSTA DE PROJETO DE MESTRADO ORIENTADOR(A)/SUPERVISOR: Helena Silva (PhD) GRUPO/GROUP: Evolutionary Genomics and Blue Biotechnology Emergent Biotechnologies and Seafood Processing (IPMA) CO ORIENTADOR(A)/CO SUPERVISOR: GRUPO/GROUP: LOCAL DE REALIZAÇÃO DO TRABALHO/PLACE OF WORK: IPMA, I.P. Algés TEMA/THEME: Molecular zoogeography of bivalve molluscs RESUMO/SUMMARY RESUMO: Numerous species of bivalves are exploited commercially for food. Their origin and distribution is of major relevance and of economic importance. Though their distribution is relatively well known in many production areas, there are still the need for easy and quick geographic identification of some species of bivalves. Some molecular biology methodologies seem promising for this purpose. Some sampling in specific areas will be needed, DNA extraction, amplification, and purification prior sequencing will be done. Morphological identification of specimens will also accompany the molecular biology work. OBJECTIVOS/OBJECTIVES: The object of this work is to explore molecular biology methodologies for characterization and geographic identification of bivalve molluscs from known production areas.

24 PRINCIPAIS METODOLOGIAS/MAIN METHODOLOGIES: Basic molecular biology techniques, PCR, Real Time PCR. Analysis and treatment of results, use of Genetic Databases like BOLD and Blast. Anatomic identification of bivalves. REFERÊNCIAS/REFERENCES: ÁREA CIENTÍFICA/SCIENTIFIC AREA: Molecular Biology/Biology

25 BLUE YOUNG TALENT PLUS BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL CIIMAR PROPOSTA DE PROJETO DE MESTRADO ORIENTADOR(A)/SUPERVISOR: Joana Soares GRUPO/GROUP: Endocrine Disruption and Emergent Contaminants (EDEC) CO ORIENTADOR(A)/CO SUPERVISOR: Miguel Machado Santos GRUPO/GROUP: Endocrine Disruption and Emergent Contaminants (EDEC) LOCAL DE REALIZAÇÃO DO TRABALHO/PLACE OF WORK: CIIMAR (Interdisciplinary Centre of Marine and Environmental Research) TEMA/THEME: Application of embryo bioassay as a high throughput approach to toxicological screening of emerging contaminants RESUMO/SUMMARY RESUMO: Presently, concerns have increased on the toxicological risk of emerging pollutants, such as hightech materials, nanoparticles, pharmaceuticals, etc. Ultimately, the aquatic ecosystems are the final destination of these compounds, that can thus threat the integrity of a vast array of organisms [Soares et al., 2018]. Many of these chemicals are not new, but the knowledge on their behaviour, toxicity as well as ecological impact is still scarce, urging the need to improve this chemicals environmental risk assessment [Santos et al., 2018]. Concomitantly, the current paradigm of toxicity testing relies on the use of extensive juvenile/animals. This approach is time-consuming, expensive, and inadequate to face the increasing need of high-throughput testing of an increasing number of new chemicals. One of the most promising alternatives is the use of embryo bioassays. Importantly, available toxicological data dealing with biologically active compounds and priority chemicals indicates that impairment in early development is expected to provoke delayed effects at the adult stage, providing useful ecotoxicological information [Soares et al., 2009] It is in this context that the current project proposal emerges. We propose to improve environmental risk assessment of emerging pollutants through the application of sensitivity embryo bioassays with aquatic animals, linking the biological effects with affected pathways.

26 OBJECTIVOS/OBJECTIVES: The following specific aims will be addressed: a) Screen emerging pollutants ecotoxicological effects using embryo bioassays, b) Develop biochemical and molecular tools to characterize the mode of action of emerging pollutants, c) Improve emerging pollutants ecotoxicological risk assessment. PRINCIPAIS METODOLOGIAS/MAIN METHODOLOGIES: Embryo zebrafish bioassays will be used as they have already been significantly validated at international levels, and already incorporate the screening of sublethal effects in key stages of embryonic development. Furthermore, our group work has a long experience on embryo development and full-life cycle studies with Danio rerio as well as of markers of development. We will select for toxicity testing representative groups of emerging pollutants that raise major concern. Embryo exposure to the toxicants will be performed through water and microinjection. For those chemicals where sufficient toxicological data is available in international literature, we will contrast the information with our results to demonstrate the sensitivity of these embryo bioassays in the hazard and risk characterization of emerging pollutants. After determining the effects of the tested chemicals on the embryonic development, the screening of the chemicals mode of action will be performed, using both biochemical and molecular tools (i.e., oxidative stress, Real time PCR, immunocitochemistry). REFERÊNCIAS/REFERENCES: Soares, J., Neuparth, T., Lyssimachou, A., Lima, D., André, A., Reis-Henriques, M.A., Castro, L.F.C., Carvalho, A.P., Monteiro, N.M., Santos, M.M., α-ethynilestradiol and tributyltin mixtures modulates the expression of NER and p53 DNA repair pathways in male zebrafish gonads and disrupt offspring embryonic development. Ecological Indicators 95(2), doi: /j.ecolind Soares, J., Coimbra, A.M., Reis-Henriques, M.A., Monteiro, N.M., Vieira, M.N., Oliveira, J.M.A., Guedes-Dias, P., Fontaínhas-Fernandes, A., Sila Parra, S., Carvalho, A.P., Santos, M.M., Disruption of zebrafish (Danio rerio) embryonic development after full life-cycle parental exposure to low levels of ethinylestradiol. Aquatic Toxicology 95(4), doi: /j.aquatox Santos, M.M., Ruivo, R., Capitão, A., Fonsexa, E., Castro, L.F.C., Identifying the gaps: resources and perspectives on the use of nuclear receptor based-assays to improve hazard assessment of emerging contaminants. Journal of Hazardous Materials 15, doi: /j.jhazmat ÁREA CIENTÍFICA/SCIENTIFIC AREA: Endocrine Disruption and Ecotoxicology

27

28 BLUE YOUNG TALENT PLUS BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL CIIMAR PROPOSTA DE PROJETO DE MESTRADO ORIENTADOR(A)/SUPERVISOR: Leonardo J. Magnoni GRUPO/GROUP: LANUCE CO ORIENTADOR(A)/CO SUPERVISOR: Francisco Guardiola GRUPO/GROUP: Nutrition and immunobiology LOCAL DE REALIZAÇÃO DO TRABALHO/PLACE OF WORK: CIIMAR TEMA/THEME: The effect of sustained swim training on the welfare of gilthead seabream (Sparus aurata) RESUMO/SUMMARY RESUMO: Swimming is a key behavioural component contributing to the fitness of most species. Several studies have shown that induced sustained swimming in cultured fish enhances growth and increases survival rates 1,2. Nevertheless, most studies showing the beneficial effects of induced swimming have been conducted on salmonid fish 3-5. However, the implementation of these conditions has frequently been overlooked in other cultured species. Reduced swimming performance of cultured Atlantic salmon versus wild counterparts has been associated with decreased disease resistance 7. Interestingly, exercise promotes health in mammals, associated with the skeletal muscle s secretory function 9. This research proposed will investigate if sustained swim training increases welfare of gilthead seabream by reducing stress and improving immune response. OBJECTIVOS/OBJECTIVES: The research proposed here will investigate if sustained swim training (SST) at optimal speeds in gilthead seabream (Sparus aurata): Improve immune function. Increase the survival and welfare by boosting robustness (reducing stress).

29 PRINCIPAIS METODOLOGIAS/MAIN METHODOLOGIES: The following tasks are proposed: Determination of the swimming capacity. Following the methodology described for other species 6. Growth trial. Fish will be housed in tanks divided into two spaces holding control or swimming groups 8. Flow will be set based on the previous test. Fish will be fed a commercial diet ad libitum twice a day. Flow will be stopped to quantify voluntary feed intake. Growth performance will be evaluated every 30 days. After 60 days half of the fish in SST groups will be transferred to 3 tanks (detraining, set at similar flow as the control). After 30 days all the groups will be sampled. Fish will be anaesthetized and blood will be collected to analyse parameters related to stress and innate immune responses. These parameters will include HCT, Hb and WBCV (blood); cortisol, lactate and glucose levels, as well as ACH50, peroxidase and lysozyme activities (plasma). Mucus will be sampled to analyze viscosity and innate immune parameters. Fish will be euthanized and head-kidney, liver and skeletal muscle will be collected. Differential expression of key genes will be assessed in tissues by qpcr, including markers of stress and immune responses (e.g. GRPs, MtHsp, IgM, ILs, TNF-α). REFERÊNCIAS/REFERENCES: 1 Palstra, A. & Planas, J. Fish under exercise. Fish Physiol. Biochem. 37, , doi:doi: /s (2011). 2 Davison, W. & Herbert, N. A. in Swimming Physiology of Fish: Towards Using Exercise to Farm a Fit Fish in Sustainable Aquaculture (eds Arjan P. Palstra & Josep V. Planas) (Springer Berlin Heidelberg, 2013). 3 Walker, M. G. & Emerson, L. Sustained swimming speeds and myotomal muscle function in the trout, Salmo gairdneri. Journal of Fish Biology 13, , doi:doi: /j tb03457.x (1978). 4 Houlihan, D. F. & Laurent, P. Effects of Exercise Training on the Performance, Growth, and Protein Turnover of Rainbow Trout (Salmo gairdneri). Canadian Journal of Fisheries and Aquatic Sciences 44, , doi: /f (1987). 5 Totland, G. K. et al. Growth and composition of the swimming muscle of adult Atlantic salmon (Salmo salar L.) during long term sustained swimming. Aquaculture 66, , doi: / (87) (1987). 6 Tudorache, C., Viaene, P., Blust, R., Vereecken, H. & De Boeck, G. A comparison of swimming capacity and energy use in seven European freshwater fish species. Ecol. Freshw. Fish 17, , doi: /j x (2008). 7 Castro, V. et al. Disease resistance is related to inherent swimming performance in Atlantic salmon. BMC Physiology 13, 1, doi: / (2013). 8 Palstra, A. P. et al. Deep RNA Sequencing of the Skeletal Muscle Transcriptome in Swimming Fish. PLoS ONE 8, e53171, doi: /journal.pone (2013). 9 Pedersen, K. A. in Comprehensive Physiology (ed R. Terjung) (2013). ÁREA CIENTÍFICA/SCIENTIFIC AREA: Biology, Aquatic sciences, Animal welfare, Animal physiology

30 BLUE YOUNG TALENT PLUS BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL CIIMAR PROPOSTA DE PROJETO DE MESTRADO ORIENTADOR(A)/SUPERVISOR: Luisa Valente GRUPO/GROUP: LANUCE CO ORIENTADOR(A)/CO SUPERVISOR: Benjamin Costas GRUPO/GROUP: NUTRIMU LOCAL DE REALIZAÇÃO DO TRABALHO/PLACE OF WORK: CIIMAR TEMA/THEME: Valorisation of agro food by products: the potential hydrolysates in functional feeds RESUMO/SUMMARY RESUMO: Protein hydrolysates were recently identified as sustainable alternatives to replace fish meal in diets for marine fish species (3,4). They are high-quality protein sources and contain bioactive peptides able to stimulate fish growth and immunity, also having antimicrobial activity. The use of these compounds can increase fish disease resistance, reducing the need for antibiotics, and contribute to a circular economy concept targeting zero waste. Size-fractioned protein hydrolysates from agro food by products will be produced by innovative methods under the project MOBFOOD ( Such hydrolysates will be included in diets for European sea bass, an important marine fish species and fed to juveniles during 3 months. At the end of the trial, the impact of the diets on the immunological status of the fish will be evaluated and compared to a commercial diet. OBJECTIVOS/OBJECTIVES: Evaluate the bioactive properties of selected protein hydrolysates and its potential as feed ingredients for European sea bass. Different hydrolysates from agro food by products will be produced by innovative methods under the project MOBFOO and fully characterised. The most appropriate ones for sea bass will be selected and included in equilibrated diets for this species. After 3 months, the immunological status of the fish fed the hydrolysates will be compared with those fed commercial feeds.

31 PRINCIPAIS METODOLOGIAS/MAIN METHODOLOGIES: - Chemical evaluation of the hydrolysate: peptide characterisation (molecular weight by fast protein liquid chromatography (FPLC), hydrophobicity by high performance LC (HPLC) and peptide sequencing by LC with mass spectrometry (LC-MS), nutritional value (amino acid composition, protein and lipid content and fatty acid profile) - In vitro bacteriostatic and bactericidal activities of hydrolysates against bacteria with impact on D. labrax diseases - In vivo experiments with juvenile D. labrax for immune status evaluation: plasma activity of lysozyme, peroxidase, ACH50 and IgM) and haematological profile (haematocrit and blood cell counting) REFERÊNCIAS/REFERENCES: 1. Cuesta, A., Meseguer, J., Esteban, M. Vet Immunol Immunopathol 2004, 101(3 4), Machado, M., Azeredo, R., Díaz Rosales, P., Afonso, A., Peres, H., Oliva Teles, A., Costas, B. Fish & Shellfish Immunol (2), Skalli, A., Zambonino Infante, J. L., Kotzamanis, Y., Fabregat, R., Gisbert, E. Aquacult Nutr 2014, 20(2), Xu, H., Mu, Y., Liang, M., Zheng, K., & Wei, Y. Aquacult Res 2017, 48(6), ÁREA CIENTÍFICA/SCIENTIFIC AREA: Biochemistry; Biological Sciences; Veterninary Sciences

32 BLUE YOUNG TALENT PLUS BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL CIIMAR PROPOSTA DE PROJETO DE MESTRADO ORIENTADOR(A)/SUPERVISOR: Luis Filipe Rangel GRUPO/GROUP: Laboratório de Patologia Animal CO ORIENTADOR(A)/CO SUPERVISOR: Maria João Santos GRUPO/GROUP: Laboratório de Patologia Animal LOCAL DE REALIZAÇÃO DO TRABALHO/PLACE OF WORK: Faculdade de Ciências da UP / CIIMAR TEMA/THEME: Influência dos parasitas coccídeos na ocorrência de outras infeções por micro e macroparasitas em peixes de interesse económico RESUMO/SUMMARY RESUMO: O desenvolvimento da piscicultura passa pelo seu sucesso económico, ou seja, por reduzir os custos e maximizar os lucros. Um dos grandes entraves ao desenvolvimento da piscicultura são as doenças, nomeadamente as parasitoses que provocam mortalidade ou retardam o desenvolvimento e o crescimento dos peixes. Os microparasitas coccídeos, por exemplo, retardam o desenvolvimento dos peixes ao reduzirem a absorção dos nutrientes. As pisciculturas compensam esse atraso com o aumento do tempo de engorda, com a consequente redução do lucro. No entanto, desconhece se que outros efeitos estes parasitas coccídeos poderão provocar na saúde dos peixes. As pisciculturas, como são sistemas fechados, potenciam o aparecimento de constantes infecções parasitárias. Para além do aumento do tempo de engorda dos peixes, os coccídeos poderão estar a contribuir para o aumento da carga parasitária com eventuais graves prejuízos. OBJECTIVOS/OBJECTIVES: O objectivo principal deste trabalho é o de avaliar se uma maior intensidade de infecção por coccídeos, e consequente menor absorção de nutrientes, torna os peixes mais susceptíveis à infeção por outros grupos de parasitas.

33 PRINCIPAIS METODOLOGIAS/MAIN METHODOLOGIES: A metodologia a utilizar será a seguinte: i) Pesquisa de micro e macroparasitas nas espécies de peixes selecionadas; ii) Determinação das prevalências e intensidade de infecção de cada espécie de parasita por hospedeiro; iii) Comparação das prevalências e intensidades de infecção de todos os parasitas em relação aos parasitas coccídeos; iv) Caracterização morfológica e molecular das espécies parasitas novas para a ciência. REFERÊNCIAS/REFERENCES: Gjurcevic, E., Kuzir, S., Bazdaric, B., Matanovic, K., Debelic, I., Marino, F., Drasner, K. and Rosenthal, B. M. (2017). New data on Eimeria dicentrarchi (Apicomplexa: Eimeriidae), a common parasite of farmed European sea bass (Dicentrarchus labrax) from the mid eastern Adriatic. Veterinary Archives 87, Molnar, K. (2006): Phylum Apicomplexa. In Fish Diseases and Disorders, Volume 1: Protozoan and Metazoan Infections (2nd ed.), (Woo, P. T. K., Ed.), CAB International, Wallingford, pp Steinhagen, D., Oesterreich, B. and Korting, W. (1997). Carp coccidiosis: clinical and hematological observations of carp infected with Goussia carpelli. Diseases of Aquatic Organisms, 30, ÁREA CIENTÍFICA/SCIENTIFIC AREA: Parasitologia / Patologia

34 BLUE YOUNG TALENT PLUS BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL CIIMAR PROPOSTA DE PROJETO DE MESTRADO ORIENTADOR(A)/SUPERVISOR: Marcos RubalGarcía GRUPO/GROUP: Coastal Biodiversity CO ORIENTADOR(A)/CO SUPERVISOR: Puri Veiga Sánchez GRUPO/GROUP:Coastal Biodiversity LOCAL DE REALIZAÇÃO DO TRABALHO/PLACE OF WORK:LBC (CIIMAR) and Lab 2.54 (Biology Department of Faculty of Sciences, UP) TEMA/THEME: Abundance, distribution and phenology of native and non indigenous canopy macroalgae along north Portugal RESUMO/SUMMARY RESUMO: Marine forests constitute the largest biogenic structures found in benthic marine systems of the world s coldwater and temperate coastal habitats (Steneck et al., 2002). Canopy forming seaweeds are ecosystem engineers that are dramatically declining worldwide. Urban and industrial pollution can cause local extinction of canopies while at a larger spatial scale, eutrophication, and global warming are the main responsibles of canopy decline. These impacts are leading to shifts in habitat structure from canopy forming species to alternative states, in the worst case to barren grounds composed of filamentous and encrusting species (Micheli et al., 2005; Connell et al., 2008). Due to the ecological importance of assemblages dominated by canopies and the decline of their populations within the past decades, it is imperative a better knowledge of their distribution abundance and fitness to develop a conservation strategy. This proposal is integrated in the project Sargaço como recurso de alto valor numa mudança global. Vale a pena investir neste recurso? com referência POCI FEDER co financed by COMPETE 2020, Portugal 2020 and the European Union through the ERDF, and by FCT through national funds.

35 OBJECTIVOS/OBJECTIVES: The aims of this proposal are: i) To explore differences on the abundance and distribution of native and non indigenous canopies ii) To explore differences on the reproductive phenology of native and non indigenous canopies iii) To explore the differences on the fitness of native and non indigenous canopies PRINCIPAIS METODOLOGIAS/MAIN METHODOLOGIES: Field sampling of abundance and distribution of the main intertidal canopy macroalgae Determination of the reproductive status of the main intertidal canopy macroalgae Determination of the fitness of the main intertidal canopy macroalgae REFERÊNCIAS/REFERENCES: CONNELL S., RUSSELL B., TURNER D., SHEPHERD A., KILDEA T., MILLER D., AIROLDI L. & CHESHIRE A., 2008 Recovering a lost baseline: missing kelp forests from a metropolitan coast. Marine ecology progress series 360: MICHELI F., BENEDETTI CECCHI L., GAMBACCINI S., BERTOCCI I., BORSINI C., OSIO G.C. & ROMANO F., 2005 Cascading human impacts, marine protected areas, and the structure of Mediterranean reef assemblages. Ecological monographs 75(1): STENECK R.S., GRAHAM M.H., BOURQUE B.J., CORBETT D., ERLANDSON J.M., ESTES J.A. & TEGNER M.J., 2002 Kelp forest ecosystems: biodiversity, stability, resilience and future. Environmental conservation 29(4): ÁREA CIENTÍFICA/SCIENTIFIC AREA:Marine Biology, Ecology, Aquatic sciences

36 BLUE YOUNG TALENT PLUS - BYTplus CENTRO INTERDISCIPLINAR DE INVESTIGAÇÃO MARINHA E AMBIENTAL - CIIMAR PROPOSTA DE PROJETO DE MESTRADO ORIENTADOR(A)/SUPERVISOR: Marisa Almeida GRUPO/GROUP: EcoBioTec CO-ORIENTADOR(A)/CO-SUPERVISOR: Sandra Ramos GRUPO/GROUP: EcoBioTec LOCAL DE REALIZAÇÃO DO TRABALHO/PLACE OF WORK: CIIMAR TEMA/THEME: Potential of lost fishing gears for adsorption of pollutants RESUMO/SUMMARY RESUMO: Fishing-related debris represents more than 50% of the weight of the floating macroplastics pollution found in marine environment. Plastic is an inert material, but environmental dynamics can induce modifications on its surface, making it suitable to adsorb pollutants. Nevertheless, chemical pollution loads in lost fishing gear (mainly composed of plastics) have not yet been sufficiently explored, being this the aim of this work. Obtained results, regarding pollutants adsorption will contribute for a better understanding of the ecological implications of fishing-related debris. OBJECTIVOS/OBJECTIVES: To evaluate, under laboratory manipulative experiments, the potential of lost gears for adsorption of organic and inorganic pollutants