Book of abstracts. Editors: Helena Peres and Aires Oliva Teles

Transcription

1 Book of abstracts Editors: Helena Peres and Aires Oliva Teles 17th March 2017 NUTRIMU Nutrition and Immunobiology Research Group - Interdisciplinary Centre of Marine and Environmental Research Terminal de Cruzeiros do Porto de Leixões Avenida General Norton de Matos, S/N Matosinhos Portugal

2 Workshop Coordinators: Helena Peres and Aires Oliva Teles Workshop Scientific Committee: Aires Oliva Teles; Ana Couto; António Afonso; Benjamin Costas; Carolina Castro; Cláudia Serra; Francisco Guardiola; Helena Peres; Inês Guerreiro; Paula Enes; Patricia Díaz-Rosales; Sergio Fernández-Boo Workshop Local Organization: Aires Oliva Teles; Alexandre F. Diógenes; Ana Amaral; Ana Basto; Ana Couto; Andreia Soares; António Afonso; Benjamin Costas; Bruno Reis; Carolina Castro; Cláudia Mota; Cláudia Serra; Filipa Fontinha; Filipe Coutinho; Francisco Guardiola; Helena Peres; Inês Guerreiro; Joana Moura; Joana Saiote; Lourenço Pinto; Marina Machado; Nicole Martins; Patricia Díaz-Rosales; Paula Enes; Rafaela Araújo Santos; Ricardo Andrade; Ricardo Pires; Rita Azeredo; Rita Matos; Rui Magalhães; Sara Martins; Sergio Fernández-Boo; Tânia Martins; Tássia Tamires Estevão Rodrigues; Thaís Cruz. ISBN:

3 The 2nd AQUAIMPROVE Workshop aimed to promote the presentation and debate of recent research advances in aquaculture-related topics in Portugal and was directed to students, researchers, academic community, and stakeholders. It followed a first AQUAIMPROVE Workshop, held in December 2016, which had more than 120 participants. From the perception of the interest of this initiative, we were encouraged to promote this second workshop, which surpassed our best expectations as it received more than 150 participants. The success of this initiative will surely motivate us at NUTRIMU- to organize the next AQUAIMPROVE Workshop, and try to keep this initiative as a regular meeting point for the scientific community and stakeholders to discuss issues related to aquaculture. Of course, the success of this workshop is directly related to the willingness of participants to attend it, but it would not be possible to organize without the pro bono dedication of the local organization committee, composed by the NUTRIMU community. To all of them, our sincere thanks. The editors 3

4 Scientific program 9:00 Registration and Poster set-up 10:00 Plenary Talk Safety and sustainability of aquaculture. Aires Oliva Teles,, Porto 10:30 Coffee break - Poster session Chair Maria José Saavedra (UTAD/) 11:00 Cryptosporidium scophthalmi-like isolate from farmed turbot (Scophthalmus maximus) J. Costa, C. Cruz, J.C. Eiras, A. Saraiva 11:15 On the Syndesmid turbellarians (Rhabdocoela: Umagillidae): a contribution based on evidence for syndesmis aethopharynx F. Cavaleiro, L. Rangel, D. Frade, J.P. Silva, M.J. Santos 11:30 Bacillus spp. as an effective strategy to inhibit bacterial fish pathogens R.A. Santos, A. Oliva-Teles, M.J. Saavedra, P. Enes, C.R. Serra 11:45 Photo-degradation of oxytetracycline in marine aquaculture: use of sunlight to prevent the emergence of bacterial resistance J.F. Leal, I.S. Henriques, A. Correia, E.B.H. Santos, V.I. Esteves 12:00 Plenary Talk Oceanus Seminar: The concept of quality in fish Giuliana Parisi, University of Florence, Italy 12:30 Lunch 14:00 Plenary Talk Aquaculture Marine dependency ratios threats or opportunities? Tiago Aires, Aquasoja, Sorgal Chair Helena Peres () 14:30 Effect of microalgae in gilthead seabream (Sparus aurata) diets S. Jorge, P. Enes, C.R. Serra, A. Oliva-Teles, A. Couto 14:45 Optimal dietary inclusion level of taurine for European sea bass (Dicentrarchus labrax) juveniles N. Martins, T. Estevão-Rodrigues, A. Diógenes, P. Diaz-Rosales, A. Oliva-Teles, H. Peres 15:00 Dietary requirement of essential fatty acids docosahexaenoic acid and eicosapentanoic acid for meagre (Argyrosomus regius) fingerlings M. Carvalho, H. Peres, R. Saleh, A. Oliva-Teles, M. Izquierdo 15:15 New lipid sources for marine fish: studies in European seabass (Dicentrarchus labrax) I. Campos, E. Matos, Luisa M.P. Valente 15:30 A decrease in water ph modulates the innate immune system and oxygen consumption of the Senegalese sole M. Machado, F. Arenas, J.C. Svendsen, R. Azeredo, R. Serradeiro, B. Costas 15:45 Coffee break - Poster session 16:30 Plenary Talk Research on nutrition at Estação Piloto de Piscicultura de Olhão towards the sustainability of meagre aquaculture P. Pousão-Ferreira, IPMA, Olhão Chair António Afonso (ICBAS/) 16:45 A practical approach for fish operational modelling based on experimental work A. Nobre, L. Valente, I. Lupatsch 17:00 Robotic vehicles for fish farming applications - an overview M. Barros, P. Neves, H. Magalhães, C. Ferreira, P. Granchinho, H. Diogo 17:15 Implementation of an experimental integrated multitrophic aquaculture system (IMTA) S. Martins, R. Magalhães, A. Oliva-Teles, H. Peres 17:30 Gonad yield and market-related traits of sea urchin from the north of Portugal F. Rocha, B. Reis, L. Baião, S. Moutinho, F. Arenas, L.M.P. Valente 17:45 Acute hypercapnia effects on the metabolism of the pacific oyster, Crassostrea gigas (Thunberg, 1793) L. Valido, M.J. Peixoto, L.F. Pereira, J.F. Gonçalves, R. Ozório, J. Machado 4

5 Plenary presentations 5

6 1 SAFETY AND SUSTAINABILITY OF AQUACULTURE Aires Oliva-Teles Departamento de Biologia, Faculdade de Ciências da Universidade do Porto, Rua do Campo Alegre, Edifício FC4, Porto, Portugal, Centro Interdisciplinar de Investigação Marinha e Ambiental, Universidade do Porto, Terminal de Cruzeiros do Porto de Leixões, Av. General Norton de Matos s/n, Matosinhos, Portugal aoteles@fc.up.pt Aquaculture production has continuously increased in the last decades and, according to FAO, nowadays, aquaculture and fisheries contribute equally with food fisheries products. As fisheries production has stabilized in the last decades, while aquaculture production is expected to steady increase, major contribution of food fish in the future will be through aquaculture. It is, therefore, a major concern to guarantee environmental sustainability, safety, and security of aquaculture. For that purpose, it is required that aquaculture products are sufficient to meet population needs; are safe to eat and safe to the environment; are produced in a sustainable way, to assure they are available now and in the future; are resilient to shocks in production chains, including the availability of ingredients for aquafeeds; are produced in a sound manner, meaning that besides meeting legislation demands are also ethically produced and meet consumers expectations. In this presentation the above aspects will be presented and major issues raised regarding each aspect will be briefly discussed. Keywords: Aquaculture; Environment sustainability; Safety; Security; Ethics. 6

7 2 THE CONCEPT OF QUALITY IN FISH Giuliana Parisi Dipartimento di Scienze Zootecniche, Università degli Studi di Firenze, Via delle Cascine 5, Florence, Italy Food quality is perceived as a global concept. Food should be primarily safe, tasty and healthy. In the case of the fish, we are faced to a matrix that is very valuable for the peculiarities that presents but it is very delicate, being subject to rapid involutional changes during the storage. In the lecture, a brief excursus of the parameters characterizing the quality of fish will be presented. The most important aspects for the market and for consumers will be briefly described. The presentation will focus on the nutritional properties and the chemical composition of fish. Particular attention will be paid on the lipid fraction, both from the quantitative and the qualitative point of view, and on its changes related to endogenous and exogenous factors (like the diet composition). The components responsible for the functional properties of fish will be also considered. The importance of freshness for maintaining and safeguarding the safety and quality characteristics of fish from farm to table will be also approached. Keywords: Aquaculture; Fish and shellfish products; Quality of seafood. 7

8 3 AQUACULTURE MARINE DEPENDENCY RATIOS THREATS OR OPPORTUNITIES? Tiago Aires SORGAL, Sociedade de Oleos e Rações, S.A., Lugar da Pardala, S. João de Ovar, Portugal tiago.aires@sojadeportugal.pt With increasing world population (prediction of 9 Billion in 2050), larger food amounts will be needed, namely fish coming from Aquaculture. Traditionally farmed fish and crustaceans were fed with feeds composed mainly of fishmeal and fish oil. The stagnation of wild fisheries imposed a limit on the availability of marine ingredients, driving Research effort towards reduction of inclusion and/or sourcing of sustainable certified marine ingredients. The aim of this presentation is to present the industry effort to reduce pressure on the marine resources and present evidence to the Consumer. A brief explanation of the major sustainability indexes in Fish farming used today is presented. Keywords: Sustainability, Fishmeal, Fish oil, Marine Dependency ratio, Fish in Fish out 8

9 4 RESEARCH ON NUTRITION AT ESTAÇÃO PILOTO DE PISCICULTURA DE OLHÃO TOWARDS THE SUSTAINABILITY OF MEAGRE AQUACULTURE Pedro Pousão-Ferreira*, Laura Ribeiro, Marisa Barata, Florbela Soares, Ana C. Mendes, Sara Castanho, Hugo Quental-Ferreira, Saavedra, M., Narcisa M. Bandarra, Maria Emília Cunha Portuguese Institute for the Ocean and Atmosphere (IPMA) Aquaculture Research Station (EPPO) Av. do Parque Natural da Ria Formosa s/n Olhão, Portugal * Europe needs to increase fish aquaculture production in order to satisfy consumer demand for fish. Moreover, it is important that fish species have potential for processing (eg: filleting, frozen, etc.) due to changes on the type of fish product selected by consumers. Therefore, research has been carried out with meagre, a fast growing fish species with high potential for production and to diversify cultured species in South Europe. Fish rearing can be enhanced when adequate feed is used. In fact, a balanced diet is essential to optimize fish growth and mitigate feed losses to environment (either as faeces or uneaten food). The aim of this presentation is to present the advances obtain for meagre at EPPO during last 6 years. Several experiments were carried out to achieve important basic information on meagre nutritional requirement, namely level of dietary protein, the adequate ratio of protein:lipid, growth performance with alternative ingredients to fish meal, growth enhancement with different levels of phospholipids and different ratios of docosahexaenoic and eicosapentaenoic acids. In addition, the adequate size for commercialization and how temperature affects food efficiency and growth performance were also evaluated. Besides the evaluation of zootechnical parameters like namely growth rate, food conversion ratio, protein accretion among others, several other analysis were performed to characterize meagre the welfare, physiology, the metabolism and sanitary condition, in order to obtain an integrated response of fish to diets. The large experience on marine aquaculture at EPPO contributed to the success of meagre as species for aquaculture production and higher growth rates and higher feed efficiency has obtained along these years. Keywords: Protein content; Protein:lipid ratio; Alternative ingredients; Phospholipids; Temperature. Research funded with AQUACOR project ( FEP-03; PROMAR) and DIVERSIAQUA project (MAR2020). To SPAROS for the experimental diets produced under NUTRICOR ( FEP-19; PROMAR). 9

10 Oral presentations 10

11 1 Cryptosporidium scophthalmi - LIKE ISOLATE FROM FARMED TURBOT (Scophthalmus maximus) Joana Costa 1, Cristina Cruz 1,2, Jorge C. Eiras 1,2 and Aurélia Saraiva 1,2* 1 Interdisciplinary Centre of Marine and Environmental Research, Av. General Norton de Matos s/n Matosinhos, Portugal, 2 Faculty of Sciences of University of Porto, Rua do Campo Alegre, Edifício FC4, , Porto, Portugal * amsaraiv@fc.up.pt Cryptosporidium is an apicomplexan parasite of high medical and veterinary importance, once it can infect humans and a wide range of mammals, birds, reptiles and fish (Ryan U, Cryptosporidium in birds, fish and amphibians. Exp Parasitol 124: ). Depending on the host, cryptosporidiosis can cause diarrheal disease in mammals, diarrhea and/or catarrhal respiratory signs in birds and gastritis in reptiles and possibly fish (O'Donoghue PJ, Cryptosporidium and cryptosporidiosis in man and animals. Int J Parasitol 25: ; Ryan U, Cryptosporidium in birds, fish and amphibians. Exp Parasitol 124: ). In this study, we provide a histological characterization of a Cryptosporidium found in the intestine of farmed turbot. Different parasitic stages were identified deep within the intestinal epithelium, and lying on apical surface of enterocytes and intestinal lumen. The deeper stages were observed inside vacuoles in hypertrophic enterocytes. Enterocyte necrosis and/or loss of tissue integrity with detachment of the epithelium from lamina propria were sometimes observed. Histological findings agree to what has been previously described for Cryptosporidium scophthalmi. Comparative genetic characterization, performed using 18S rrna and actin loci of the turbot Cryptosporidium, revealed that this isolate was different from all other species/genotypes already described. Keywords: Cryptosporidium; Turbot; Histology; Aquaculture. This work was partially funded by the Project AQUAIMPROV (reference NORTE FEDER ), cofinanced by the North Portugal Regional Operational Programme (ON.2 O Novo Norte), under the National Strategic Reference Framework (NSRF), through the European Regional Development Fund (ERDF). 11

12 2 ON THE SYNDESMID TURBELLARIANS (RHABDOCOELA: UMAGILLIDAE): A CONTRIBUTION BASED ON EVIDENCE FOR SYNDESMIS AETHOPHARYNX Francisca Cavaleiro*, Luís Rangel, Duarte Frade, José Pedro Silva, Maria João Santos Universidade do Porto, Faculdade de Ciências, Departamento de Biologia, Rua do Campo Alegre, s/n, Edifício FC4, Porto, Portugal Centro Interdisciplinar de Investigação Marinha e Ambiental (/CIMAR), Terminal de Cruzeiros do Porto de Leixões, Avenida General Norton de Matos, s/n, Matosinhos, Portugal * fcavaleiro@fc.up.pt The syndesmid turbellarians are platyhelminths (umagillid rhabdocoels) which live as endosymbionts in echinoid hosts, namely, sea urchins and sand dollars. Presently, the body of knowledge on them is still very fragmentary, the available information mostly respecting species morphology as revealed by light microscopy analysis. In this work, a species of syndesmid found in the sea urchin Paracentrotus lividus (Lamarck, 1816) i.e., Syndesmis aethopharynx Westervelt & Kozloff, 1990 is revisited based on light microscopy, scanning electron microscopy (SEM) and molecular phylogenetic analysis. Also, species ecological data are considered for analysis. The SEM analysis allowed to prove that the anterior-ventral region of the body tegument is equipped with cilia, thus supporting the idea that the non-ciliated epidermal region observed in some species of umagillids represents an apomorphy. Cilia were observed to be long and to culminate in an enlarged, round tip, which indicates that they play a major role in locomotion. The results of the molecular phylogenetic analysis suggest that syndesmids are closely-related to symbionts of other echinoderms (i.e. holothurians) and provorticids. Moreover, they suggest that syndesmids occupy a position between intestinal and coelomic symbionts. This is an interesting finding as, in this study, both the digestive tube and perivisceral fluid were recorded as sites of infection. The recorded infection levels were low and the symbiont was observed to be aggregated in its host population. This probably reflects the aggregated distribution of the sea urchins in the macroenvironment and the fact that S. aethopharynx is a very delicate symbiont founding harsh macroenvironmental conditions while dispersing between intertidal pools. Keywords: Syndesmid turbellarians; Paracentrotus lividus; Syndesmis aethopharynx; Revisitation. This work was partially supported by the Structured Program of R&D&I INNOVMAR - Innovation and Sustainability in the Management and Exploitation of Marine Resources, reference NORTE FEDER , namely within the Research Line INSEAFOOD Innovation and valorization of seafood products: meeting local challenges and opportunities, within the R&D Institution (Interdisciplinary Centre of Marine and Environmental Research), supported by the Northern Regional Operational Programme (NORTE2020), through the European Regional Development Fund (ERDF). A word of gratitude is due to the working teams of Professor Adriano Bordalo e Sá and Doctor Francisco Arenas, which collected the samples of oyster and sea urchin for analysis. 12

13 3 Bacillus spp. AS AN EFFECTIVE STRATEGY TO INHIBIT BACTERIAL FISH PATHOGENS Rafaela A. Santos 1*, Aires Oliva-Teles 1,2, Maria José Saavedra 2,3,4, Paula Enes 2 and Cláudia R. Serra 2,4 1 Departamento de Biologia, Faculdade de Ciências, Universidade do Porto, Portugal. 2 CIMAR/ - Centro Interdisciplinar de Investigação Marinha e Ambiental, Matosinhos, Portugal 3 Departamento de Ciências Veterinárias, ECAV, Universidade de Trás-os-Montes e Alto Douro, Vila Real, Portugal. 4 CECAV-Centro de Ciência Animal e Veterinária, Universidade de Trás-os-Montes e Alto Douro, Vila Real, Portugal. * up @fc.up.pt Bacterial diseases in aquaculture have been increasing around the world, limiting the expansion of the sector. The conventional approach to avoid bacterial outbreaks was, and still is in many regions of the world, the massive use of in-feed antibiotics, which exacerbates the potential for antimicrobial resistance development among pathogens. Nowadays, new disease-preventive measures are emerging, such as the use of probiotics, in particular Bacillus spp., known to produce Natural Antimicrobial Compounds (NACs). These NACs help their Bacillus producers to constitute a barrier to pathogens proliferation, contributing to host health. Taking advantage of Bacillus ubiquitous and sporeforming nature, we were able to isolate, identify, and characterize from the gut of aquaculture fish species (European sea bass, gilthead sea bream and white sea bream) several strains with antimicrobial activity against important bacterial fish pathogens. A total of 176 isolates representing different colony morphologies and samples were obtained from fish intestinal contents, and an identification based on the 16S rrna gene revealed a clear abundance of B. subtilis. Screening for NACs production revealed that 52% of all isolates displayed antagonistic activity. Of these, six were selected as most promising isolates, capable of inhibiting bacterial growth of Aeromonas spp., Vibrio spp., Photobacterium sp., and Staphylococcus sp. By characterizing the localization (intra- or extra-cellular) of the inhibitory molecules, it was observed that cell-free supernatants of three out of the six selected isolates significantly inhibited growth of each pathogen tested, except for A. salmonicida. Interestingly, by characterizing for anti-biofilm activity, it was shown that the same three isolates significantly reduced the adherence of A. salmonicida. This suggests the possibility for an alternative fighting measure against this important fish pathogen. The corresponding bioactive molecules are currently under further characterization to evaluate their potential for the treatment of fish infections in aquaculture. Keywords: Aquaculture; Fish pathogens; Sporeformers; Natural Antimicrobial Compounds. Acknowledgments P.Enes and C.R.Serra were supported by grants SFRH/BPD/101012/2014 and SFRH/BPD/101038/2014 respectively, from FCT (Portuguese Science Foundation). The authors thank F.Tavares (CIBIO-InBIO), B. Costas (), MA Morinigo (Univ. Málaga) for the gift of pathogenic bacterial strains, and A.Henriques (ITQB-NOVA) for the gift of B. subtilis

14 4 PHOTO-DEGRADATION OF OXYTETRACYCLINE IN MARINE AQUACULTURE: USE OF SUNLIGHT TO PREVENT THE EMERGENCE OF BACTERIAL RESISTANCE Joana F. Leal 1*, Isabel S. Henriques 2, A. Correia 2, Eduarda B.H. Santos 1, Valdemar I. Esteves 1 1 Departamento de Química & CESAM, Universidade de Aveiro, Campus Universitário de Santiago, Aveiro, Portugal 2 Departamento de Biologia & CESAM, Universidade de Aveiro, Campus Universitário de Santiago, Aveiro, Portugal * joanaleal@ua.pt Oxytetracycline (OTC) is a broad spectrum antibiotic used for treatment and control of a wide variety of bacterial infections, including in aquaculture. The main concern related to the use of antibiotics is the emergence of bacterial resistance. Some authors refer that about % of antibiotics administered with fish food in aquaculture are excreted into water [1], contributing to contamination of aquatic systems, which can act as a reservoir of resistance genes [2]. Aquaculture water samples were spiked with OTC and subjected to several irradiation experiments, using simulated sunlight with an irradiance of 550 W/m 2. OTC was quantified by HPLC-DAD. The overall quantum efficiency of photoreactions, as well as, the outdoor half-life times of OTC in water, at 40 degrees north of latitude (at sea level) were predicted. The results obtained are very promising to use the photo-degradation for remediation of marine aquaculture s waters. The outdoor half-life times estimated for the brackish aquaculture s waters, spiked with OTC 4 mg/l, range between 21 and 26 minutes, for a midsummer day [3]. The well-diffusion method was chosen to perform the antibacterial assays, before and after OTC photo-degradation. The bacterial strains considered for these experiments were Escherichia coli, Vibrio sp. and Aeromonas sp. It was observed a decrease of the biological activity with the increase of the irradiation time, suggesting that, in the conditions tested, the OTC by-products do not retain antibacterial activity after irradiation under sunlight [4]. Keywords: Antibiotics; Marine aquaculture; Photo-degradation; Sunlight; Antibacterial activity. References [1] S. Gastalho, G.J. dasilva, F. Ramos, Acta Farmacêutica Portuguesa, 3 (2014): [2] M. Tacão, A. Correia, I. Henriques, Applied and Environmental Microbiology, 78 (2012): [3] J.F. Leal, V.I. Esteves, E.B.H. Santos, Environmental Pollution 213 (2016): [4] J.F. Leal, I.S. Henriques, A. Correia, E.B.H. Santos, V.I. Esteves, Environmental Pollution 220 (2017): Authors would like to acknowledge funding from National Foundation for Science and Technology FCT (POPH/FSE) to Centre for Environmental and Marine Studies (CESAM) (UID/AMB/50017/2013) and ibimed (UID/BIM/04501/2013). Isabel Henriques thanks FCT for her contract (FCT Investigator Programme IF/00492/2013). Joana F. Leal thanks FCT for her PhD grant (SFRH/BD/88572/2012). 14

15 5 EFFECT OF MICROALGAE IN GILTHEAD SEABREAM (Sparus aurata) DIETS S. Jorge 1 *, P. Enes 2, C. R. Serra 2, A. Oliva Teles 1,2 and A. Couto 2 1 Departamento de Biologia, Faculdade de Ciências da Universidade do Porto, Rua do Campo Alegre s/n, Edifício FC4, Porto, Portugal 2 Centro Interdisciplinar de Investigação Marinha e Ambiental (/CIMAR), Universidade do Porto, Terminal de Cruzeiros do Porto de Leixões, Av. General Norton de Matos s/n, Matosinhos, Portugal * sarasijorge@gmail.com Aquafeeds for carnivorous fish, traditionally demand high amounts of fishmeal and fish oil. However, the increasing demand and prices of these ingredients, combined with the current state of the worldwide fish stocks FAO (2014), prompted the search for alternatives. Ingredients derived from common terrestrial crops are already largely used in aquafeeds as marine-derived substitutes, but often present antinutritional factors that can negatively affect fish performance. Microalgae are a promising feed ingredient for aquafeeds, as source of protein, lipids (particularly, ω6 and ω3 fatty acids), minerals and vitamins (Reyes-Becerril et al., 2013), but also of bioactive compounds with immunostimulant and health-promoting effects (Cerezuela et al., 2013). In the present work, a short-term trial was performed toevaluate Nannochloropsis sp. as functional ingredient (0, 0.5, 0.75 and 1.5% inclusion) in lowfishmeal diets for seabream (Sparus aurata). Microalgae effect was evaluated on growth, feed utilization, intestinal morphology, function and microbiota. Zootechnical performance tended to improve with increasing levels of dietary Nannochloropsis. Digestive function and morphology were unaffected by microalgae inclusion, although increased observations of eosinophilic granular cells in the intestine hinted an immune reaction. Inclusion of Nannochloropsis resulted in increased operation taxonomical units, suggesting microbiota modulation. Keywords: Aquafeeds; Microalgae; Functional ingredient; Gilthead seabream; Nannochloropsis sp. Project ALGAFISH (PTDC/MAR-BIO/6233/2014-POCI FEDER016796), supported by Fundação para Ciência e a Tecnologia and co-financed by Fundo Europeu de Desenvolvimento Regional (FEDER), fromcompete Programa Operacional Competitividade e Internacionalização (POCI). Partner Buggypower, Lda kindly supplied microalgae biomass 15

16 6 OPTIMAL DIETARY INCLUSION LEVEL OF TAURINE FOR EUROPEAN SEA BASS (Dicentrarchus labrax) JUVENILES N. Martins 1*, T. Estevão-Rodrigues 1, A. Diógenes 1,2, P. Diaz-Rosales 2, A. Oliva- Teles 1,2, H. Peres 1,2 1 Department of Biology, Faculty of Sciences University of Porto, Portugal 2 Interdisciplinary Centre of Marine and Environmental Research (-UP), Porto, Portugal * nicolemartins18@hotmail.com The aim of this study was to evaluate the optimal dietary inclusion level of taurine in diets for European sea bass juveniles, by assessing fish growth performance and some physiological effects related to taurine metabolism. Eight isoproteic (45% crude protein) and isolipidic (18% crude lipid) diets were formulated to contain a mixture of plant feedstuffs and fish meal (corresponding to 80% and 20% of total dietary protein, respectively) and taurine (Tau) included at levels ranging from 0 to 1.5% (diets 0Tau; 0.1Tau; 0.2Tau; 0.3Tau; 0.5Tau; 0.7Tau; 1Tau; 1.5Tau). Triplicate groups of fish were fed these diets by hand, twice a day, 6 days a week, until visual apparent satiation for 10 weeks. Growth performance increases with dietary taurine level up to 0.5% and then slightly decreased. Feed intake and feed efficiency were not affected by dietary taurine level. Dietary treatments did not affect whole body composition, except for lipid content that decreased as dietary taurine level increased. Nitrogen retention, expressed as g/kg body weight/day or as % nitrogen intake, was not affected by dietary treatments. Post-prandial plasma total cholesterol, HDL, LDL, total and indirect bilirubin were affected by treatments, suggesting a taurine role in lipid metabolism. A five-parameter saturation kinetics model was used to fit daily growth index and nitrogen retention (g kg ABW-1 day-1) against dietary taurine levels. Based on that model, the optimal dietary taurine inclusion was 0.37% both for maximum growth and nitrogen retention Keywords: Taurine requirements; Plant feedstuffs; European sea bass; Growth performance. This work was supported by the Structured R&D&I Project INNOVMAR - Innovation and Sustainability in the Management and Exploitation of Marine. Resources (ref. NORTE FEDER ) within the research line "INSEAFOOD - Innovation and valorization of seafood products: meeting local challenges and opportunities", founded by the Northern Regional Operational Programme (NORTE2020) through the European Regional Development Fund (ERDF). 16

17 7 DIETARY REQUERIMENT OF ESSENTIAL FATTY ACIDS (DOCOSAHEXAENOIC ACID AND EICOSAPENTAENOIC ACID) FOR MEAGRE (Argyrosomus regius) FINGERLINGS M. Carvalho 1*, H. Peres 1,2, R. Saleh 3, A. Oliva-Teles 1,2, M. Izquierdo 3 1 Department of Biology, Faculty of Sciences, University of Porto, Portugal 2 Interdisciplinary Centre of Marine and Environmental Research (-UP, Porto, Portugal) 3 Grupo de Investigación en Acuicultura, University of Las Palmas de Gran Canaria, Gran Canaria, Spain * martaribeirocarvalho27@gmail.com Fatty acids play key biological roles in marine fish, being required to sustain growth, development and survival. For new aquaculture species, the knowledge regarding their specific dietary requirements is scarce and the establishment of well balance diets that meet the nutrient requirement of the species is important to optimize a large-scale production. This is the case of meagre (Argyrosomus regius), a promising new aquaculture species, with great potential due to their high growth rate, feed efficiency, flesh quality, market prices and easy adaptation to captivity. Therefore, the present study aimed to evaluate the essential fatty acid (EFA) requirements of meagre fingerlings, namely DHA and EPA. A feeding trial was performed at FCPCT (Fundación Canaria Parque Científico Tecnológico, Las Palmas de Gran Canaria) testing 6 increasing EFA levels: 0.8, 1.2, 1.7, 2.6, 3.3 and 3.6%. Triplicate groups of meagre fingerlings with an initial body weight of ±2.8g were fed each experimental diet by hand, three times a day, until apparent visual satiety, 6 days a week, for 4-weeks. During the trial the water temperature was kept constant at 23-24ºC. Fish fed 0.8% HUFA obtained the lowest growth. Increasing dietary % EFA improved the specific growth rate and feed conversion ratio, and % HUFA in diet above 2.6% reduced significantly the % of lipids in liver. Dietary levels of 3.3% HUFA improved the fatty acid profile in fish, increasing DHA, EPA and ARA contents. Finally, fish fed diets with more than 2.6% HUFA obtained lower steatosis in liver. The results suggested that the dietary requirement of EFA for optimal growth recommended for meagre fingerlings is 2.2 and 2.5% based on length and weigh, respectively. Keywords: Meagre fingerlings; Essential fatty acids; Docosahexaenoic acid; Eicosapentaenoic acid; Requirement. This work was part of the European project DIVERSIFY- Exploring the biological and socio-economic potential of new/emerging candidate fish species for expansion of the European aquaculture industry, funded under the 7th Framework Programme of the European Commission (7FP-KBBE-2013). M. Carvalho was financed by the University of Porto, under the mobility programme ERASMUS+. 17

18 8 NEW LIPID SOURCES FOR MARINE FISH: STUDIES IN EUROPEAN SEABASS (Dicentrarchus labrax) I. Campos 1,2*, E. Matos 3, Luisa M.P. Valente 1,2 1 -CIMAR L.A., Centro Interdisciplinar de Investigação Marinha e Ambiental, Matosinhos, Portugal; 2 ICBAS, Instituto de Ciências Biomédicas de Abel Salazar, Universidade do Porto 3 SORGAL, Sociedade de Óleos e Rações, S.A., Estrada 109 Lugar da Pardala, S. João OVAR, Portugal * bioines@gmail.com The unsustainability of fish oil use in aquafeeds raises ecological and ethical concerns, while rendered animal fats, obtained from animal by-products, have low price and wide availability, being interesting alternatives to fish oil. Their higher content in saturated fatty acids (SFA) and lower content in polyunsaturated fatty acids (PUFA) can impair digestibility, growth performance and final product quality. However, the high SFA and low PUFA contents make these ingredients more resistant to auto-oxidation than fish oil (Watanabe 1989). The present study was conducted to characterize and evaluate the effects of three locally produced rendered animal fats (fish oil, FO; poultry fat, PF; and a mammal fat mixture, MF 70% lard and 30% beef tallow) on the in vivo apparent digestibility of fatty acids in European seabass. A commercial-based diet was used to create isoenergetic (23 kj/g) and isonitrogenous (48% crude protein) diets, by incorporating FO, PF or MF as lipid source at 14% of each diet. 5-α cholestane was added as inert digestibility marker. Each diet was fed to quadruplicate groups of 100 g European seabass. The fatty acid digestibility results obtained were high regardless of the fatty acid source, but the omega-3 fatty acids digestibility values were highest in the diet with fish oil. Land animal fats, with higher levels of saturated and monounsaturated fatty acids, had reduced digestibility of polyunsaturated fatty acids, even though this was only significant in the PF diet. These results suggest that all the tested rendered animal fats could be good energy sources but the lower ADC values of omega-3 fatty acids in land animal fats may affect fish flesh quality. Keywords: European seabass; Fatty acids digestibility; Animal fats; Local by-products; Circular economy. References Watanabe, T. (1989). "Nutritive value of animal and plant lipid sources for fish." In: Progress in Fish Nutrition. Proc. Fish Nutrition Symp. (Ed.) Shiau, S.Y. Marine Food Science Series, 9: I. Campos was financially supported by Fundação para a Ciência e Tecnologia, Portugal, and Soja de Portugal, through the grant PDE/BDE/113668/2015. This work was subsidized by Project ANIMAL4AQUA, funded by Portugal 2020, financed by the European Regional Development Fund (FEDER) through the Operational Competitiveness Program (COMPETE) reference POCI FEDER

19 9 A DECREASE IN WATER PH MODULATES THE INNATE IMMUNE SYSTEM AND OXYGEN CONSUMPTION OF THE SENEGALESE SOLE Marina Machado 1,2*, Francisco Arenas 1, Jon C Svendsen 1,5, Rita Azeredo 1,3, Renata Serradeiro 4, Benjamín Costas 1,2 1 Centro Interdisciplinar de Investigação Marinha e Ambiental (), Universidade do Porto, Portugal. 2 Instituto de Ciências Biomédicas Abel Salazar (ICBAS-UP), Universidade do Porto, Portugal. 3 Departamento de Biologia, Faculdade de Ciências da Universidade do Porto (), Porto, Portugal. 4 Sea8- Aquacria Piscícolas, SA, Rua do Lavadouro, Quintas do Norte, Torreira, Portugal 5 National Institute of Aquatic Resources (DTU Aqua), Technical University of Denmark, Jægersborg Allé 1, DK Charlottenlund, Denmark * mcasimiro@ciimar.up.pt High production densities is a feature of intensive recirculation aquaculture production systems and may lead to a decrease of ph by the accumulation of water CO 2 [1]. The effects of hypercapnia rely on CO 2 concentration and duration of fish exposure [2] and, while being quoted as a stress factor, hypercapnia can also be responsible for the deviation of metabolic energy [3] and alteration of basal energy requirements [2]. Thus, this study intended to assess the effects of acute and prolonged exposure to low ph levels on the innate immune mechanisms and oxygen consumption for Senegalese sole (Solea senegalensis). In four independent recirculating seawater systems, a ph-stat system (Aqua Medic, AT Control) was used to handle the flux of CO 2 from a reservoir into the water establishing four ph levels: 8.2 (control), 7.9, 7.6 and 7.3. Twelve fish were sampled at 4, 24 hours and 4 weeks after the beginning of the trial in order to assess acute and prolonged effects of a decrease in ph. The haematological profile and plasma humoral immune parameters were evaluated. Finally, a respirometry system in combination with metabolic chambers was used to record oxygen consumption in resting and stressed fish at the pretended ph. The hypothesis that hypercapnia exposure can be stressful is demonstrated with the activation of some humoral mechanisms and increase of peripheral blood cells upon acute and chronic exposure to low ph. Despite that, maximum and standard metabolic rates were not altered while a tendency for the reduction of the aerobic scope shows a gradual reduction of the oxygen range available for the normal metabolism. Keywords: Hypercapnia; Flatfish; Immune cells; Humoral immune parameters; Respirometry. References 1) Summerfelt, S.T., et al.(2000) Aquaculture Engineering 22, ) Santos, G.A., et al. (2013) Aquaculture Research 44, ) Cech, J.J. et al. ( 2002) J. Appl.Ichth. 18, This work was partially funded by projects AQLARA ( FEP-0034) supported by PROMAR Program (Portugal) and PEst-C/MAR/LA0015/2013 by the European Regional Development Fund (ERDF) through the COMPETE and POPH Programmes and national funds through Foundation for Science and Technology (FCT). M. Machado, A. Couto R. Azeredo, J.C. Svendsen and B. Costas were supported by FCT(SFRH/BD/108243/2015, SFRH/BPD/101354/2014,SFRH/BD/89457/2012, SFRH/BPD/89473/2012 and IF/00197/2015, respectively). 19

20 10 A PRACTICAL APPROACH FOR FISH OPERATIONAL MODELLING BASED ON EXPERIMENTAL WORK Ana Nobre 1*, Luísa Valente 1,2, Ingrid Lupatsch 3 1 CIMAR/ Centro Interdisciplinar de Investigação Marinha e Ambiental, Terminal de Cruzeiros do Porto de Leixões, Avenida General Norton de Matos, S/N, Matosinhos, Portugal 2 ICBAS Instituto de Ciências Biomédicas de Abel Salazar, Universidade do Porto, Rua Jorge Viterbo Ferreira 228, Porto, Portugal. 3 AB Agri Ltd Associated British Agriculture, 64 Innovation Way, PE2 6FL, Peterborough, United Kingdom. * anobre@ciimar.up.pt Optimization practices are crucial to aquaculture sustainability. Farmers need to control raw resources, efficiency, variables affecting it, disease outbreaks, waste production and real-time management of the process. A wide range of published models exist which have the potential to help understanding, predicting and managing the biological processes of an aquaculture production system. For the operational simulation of farm production, the advantages of a very detailed model might dissipate with the level of uncertainty of data required for its setup. Bioenergetic models are potential candidates given these provide a simple overview of the main processes for fish growth simulation which corresponds to an energy mass balance. Herein we present a generalization of the bioenergetics modelling approach linked with standard experiments for the setup/calibration for individual growth. The modelling approach presented herein is a trade-off between detailed process simulation and feasibility of implementation and data gathering on an operational setting to provide real-time distilled information for farm managers. We exemplify the model implementation for gilthead seabream based on the set of growth and metabolic experiments developed by Lupatsch (2003), Lupatsch et al. (2003) and Oliva-Teles et al. (2011) for the factorial approach. Keywords: Fish modelling; Bioenergetics; metabolism and growth experiments; model calibration. References Lupatsch I, PhD thesis. Universitäts-und Landesbibliothek Bonn. Lupatsch I, Kissil G, Sklan D, The Israeli Journal of Aquaculture Bamidgeh 55(4): Oliva-Teles, A, Lupatsch, I, Nengas, I, In Sparidae: Biology and Aquaculture of Gilthead Sea Bream and Other Species. Wiley-Blackwell, pp ISBN: Financial support was provided by the Portuguese Foundation for Science and Technology (FCT) as Postdoc scholarship to Ana Nobre (SFRH/BPD/109442/2015). 20

21 11 ROBOTIC VEHICLES FOR FISH FARMING APPLICATIONS - AN OVERVIEW Manuel Barros 1, Pedro Neves 1, Hugo Magalhães 2, Carlos Ferreira 1, Pedro Granchinho 1 and Hugo Diogo 3 1 Instituto Politécnico de Tomar, fmbarros@ipt.pt, granchinho@ipt.pt, 2 TagusValley, hugo.magalhães@taggusvalley.pt, 3 Compta Emerging Business, hugo.diogo@compta.pt Industrialization in the aquaculture sector is associated with the introduction of technology, since a large number of parameters have to be controlled in modern aquaculture systems. Some of these operations require sophisticated tools and specially designed facilities that have evolved through intensive research and great innovation (Mustafa 2016). Some of the custom-made technological inventions for aquaculture operations include, for example, the creation of semi-submersible cages, the implementation of automatic feeders and water recirculation systems. The use of robotics has increasingly found space among applications in the aquatic environment. The main objective is to collect information about the environment and, consequently, to manage resources better (Borović 2011). There have been reports of the use of robotic vehicles in aquaculture in applications such as: monitoring of water quality parameters, reduction of biological pests or unwanted predators and other agricultural and aquaculture applications. Boats, underwater vehicles and autonomous airplanes were designed and built to monitor and potentially manage aquaculture facilities, natural water bodies and drinking water (Dunbabin, 2009). The visible benefit of this technology is that it takes aquaculture systems to the next level, from the application of computer control and Artificial Intelligence to a greater degree of automation, effective management and decision making. The present work reviews some of the most recent robotic vehicles applied in fish farming applications and discusses its advantages and limitations. Keywords: Aquaculture, Control and Automation, Autonomous Vehicles, Robotics, Information and Communications Technology (ICT), Water Quality Monitoring. References Mustafa, F.H., A Review of Smart Fish Farming Systems. J. Aqua. Eng. Fish. Res. 2(4): Borović, B., Vasilijević, A, Kuljača, O.,2011. Potentials of Using Underwater Robotics for Fishing and Fish Farming. in Proceedings of the 10 th international workshop. Dunbabin, M., Grinham, A, Udy, J., An Autonomous Surface Vehicle for Water Quality Monitoring. pp. 2 4 in Australasian Conference on Robotics and Automation. This research is supported by Portugal 2020 AQUATROPOLIS project. The AQUATROPOLIS - Intelligent Management System for Sustainable Aquaculture is an Incentive System for Research and Technological. Development (SI I&DT) project in cooperation with the following entities: COMPTA - Emerging Business, S.A.; ALGAPLUS-Production and Commercialization of Algae and its Derivatives Lda; DOMATICA - Global Solutions, S.A.; Polytechnic Institute of Leiria (IPL); Polytechnic Institute of Tomar (IPT) and TAGUSVALLEY Association for the Promotion and Development of the Tecnopolo Valley of the Tagus Valley. The main objective of the Aquatropolis project-intelligent Management System for Sustainable Aquaculture is to develop a disruptive solution for an intelligent, optimized and automated management of aquaculture operations, in order to promote thesustainable development of the aquaculture industry in the countries of the Atlantic region. 21

22 12 IMPLEMENTATION OF AN EXPERIMENTAL INTEGRATED MULTITROPHIC AQUACULTURE SYSTEM (IMTA) Sara Martins 2, Rui Magalhães 1,2, Aires Oliva-Teles 1,2, Helena Peres 1,2* 1 CIMAR/ Centro Interdisciplinar de Investigação Marinha e Ambiental, Matosinhos, Portugal 2 Departamento de Biologia, Faculdade de Ciências, Universidade do Porto Portugal * pereshelena@ciimar.up.pt The objective of this study was to evaluate the effect of low, intermediate and high fish stoking density (LD, ID, HD, respectively) on sea urchin and seaweed productivity in an integrated multitrophic aquaculture system (IMTA). Six independent experimental scale IMTA systems were implemented at the Marine Zoological Station, Porto University, to include a 60L-unit of marine carnivorous fish (European seabass, Dicentrarchus labrax), a 60L-unit of grazing invertebrate (sea urchin, Paracentrotus lividus) and a 60Lunit of seaweed (Ulva lactuca). Each system was composed by 4 tanks in a closed recirculation system, each one supplied by a continuous flow of water from the previous tank. The fish tank was designed to include a particulate organic matter trap to ensure that all feces and unheated feed continuously moved to the sea urchin tank, and then to the seaweed tank. The experimental trial was conducted for 70 days at 18 C with a 12/12 hours light cycle. At the beginning of the trial fish stoking density was established at 3.48, 6.93 and kg/m 3 ; sea urchin density was kept constant at 19.5 kg/m 3 ; Ulva was also kept constant at 1.92 kg/m 3. Fish were daily fed with a commercial diet (42% crude protein; 18% crude fat) and sea urchin were fed two times a week with seaweed produced in the IMTA system. At the beginning and at the end of the trial, fish were group weighted and sea urchin diameter was measured. Algae were weighted at the beginning of the trial and every 7 days during the trial. At the end of the trial, there were no differences among fish groups in growth performance and feed utilization. Sea urchin produced at the ID had a higher total diameter gain. Relative growth rate of seaweed was not affected by fish density. In conclusion, fish effluents proved suitable for sea urchin and Ulva lactuca cultivation, suggesting that production and economic diversification of industrial fish facilities may be feasible. Keywords: Integrated multitrophic aquaculture system; European seabass; Sea urchin; Seaweed. This work was supported by the Structured R&D&I Project INNOVMAR - Innovation and Sustainability in the Management and Exploitation of Marine Resources (ref. NORTE FEDER ) within the research line "INSEAFOOD - Innovation and valorization of seafood products: meeting local challenges and opportunities", founded by the Northern Regional Operational Programme (NORTE2020) through the European Regional Development Fund (ERDF).We would like to express our thanks to P. Correia for the assistance during the trial. Magalhães, R. was supported by a INSEAFOOD project grant (ref. INSEAFOOD/BI/L2/ ). 22

23 13 GONAD YIELD AND MARKET-RELATED TRAITS OF SEA URCHIN FROM THE NORTH OF PORTUGAL Filipa Rocha 1*, Bruno Reis 1, Luís Baião 1, Sara Moutinho 1, Francisco Arenas 1, Luísa M.P. Valente 1 1, Interdisciplinary Centre of Marine and Environmental Research of the University of Porto * frocha@ciimar.up.pt Sea urchin gonads (roe) are considered a prized and gourmet seafood product worldwide. In Portugal, Paracentrotus lividus is most harvested in the north region, where landings reached the 28 tonnes in 2015 (INE, 2015). However, this prized resource is not used to boost our national economy, instead is being diverted to supply international markets, such as Spain. The aim of this study was to characterize sea urchins from the north coast (Praia Norte, Viana do Castelo) over 12 months in terms of (i) maximum gonad yield and (ii) seasonal variations of important traits for roe marketing, such as color (using a colorimeter), total pigments and instrumental texture (hardness and resilience, using a texture analyzer). Results showed that P. lividus has an annual reproductive cycle, with the highest gonadosomatic index (GSI) occurring in February (12%) and the lowest in June (5%). The total pigments content showed a seasonal variation, with the highest level being observed after April (start of spawning season). Gonad firmness increased by the end of the spawning season, when a concomitant increase of the GSI was observed. Resilience and color (lightness, redness and yellowness) of gonads did not significantly differ between the Summer and Autumn seasons. These outcomes suggest that the best period for harvesting high quality gonads in the Portuguese NE coast is from January up to April when gonads are mature and prior to spawning. Keywords: Sea urchin; Gonads; Seasonal; Characterization. This work was supported by the Structured R&D&I Project INNOVMAR Innovation and Sustainability in the Management and Exploitation of Marine Resources (ref. NORTE FEDER ) within the research line "INSEAFOOD - Innovation and valorization of seafood products: meeting local challenges and opportunities", founded by the Northern Regional Operational Programme (NORTE2020) through the European Regional Development Fund (ERDF). References: INE, I.P. (2015). Capturas nominais de pescado (T) e ( ) por Porto de descarga e Espécie; disponível em: [ consultado em: 14/06/

24 14 ACUTE HYPERCAPNIA EFFECTS ON THE METABOLISM OF THE PACIFIC OYSTER, Crassostrea gigas (THUNBERG, 1793) Lia Valido 2, Maria João Peixoto 1, Luís Ferreira Pereira 1, José F. Gonçalves 2, Rodrigo Ozório 1, Jorge Machado 2* 1 ICBAS Instituto de Ciências Biomédicas de Abel Salazar, Universidade do Porto, Rua Jorge Viterbo Ferreira 228, Porto, Portugal 2 Interdisciplinary Centre of Marine and Environmental Research of the University of Porto, Terminal de Cruzeiros do Porto de Leixões, Avenida General Norton de Matos, Matosinhos Portugal * jmachado@icbas.up.pt According to the Intergovernmental Panel on Climate Change (IPCC), it is estimated that the partial pressure of CO 2 (pco 2) in seawater will further increase from the current 356 μatm to 800 μatm by As the dissolved inorganic carbon dioxide increases, the equilibria will lead to higher H + and lower CO 3 2- concentrations. Thus, ocean acidification may affect biomineralization, respiratory activity and energy metabolism. The latter can be shaped directly, via intracellular ph signaling, and/or indirectly, via the elevated energy demands of acid-base and ion homeostasis. Consequently, this may lead to a trade-off between different biological processes, including homeostasis, growth and development. There are several well established methods to determine oxygen consumption rates (MO 2), among which the Automated Intermittent Flow Respirometry is considered as the most detailed and reliable. Furthermore, MO 2 is known as a proxy to metabolic rate. This research was conducted to determine whether exposure of Pacific oyster, Crassostrea gigas, to hypercapnia (0,081 kpa CO 2, seawater ph 7.5) may alter the oxygen consumption (MO 2, mgo2/kg/hr) and standard metabolic rate (SMR, mgo2/kg/hr). The analyses of the experimental data from hypercapnic and normocapnic oysters (0,036 kpa CO 2, seawater ph 8.1) are under progress, and will be presented during the AQUAIMPROVE workshop. Keywords: Acute hypercapnia; Pacific oyster; Oxygen consumption; Standard Metabolic Rate. This study is partially supported by the INSEAFOOD project, funded by NORTE 2020 through FEDER and aims to enhance scientific competences in the area of Aquaculture and Seafood Quality, which are relevant for the implementation of the North Portugal Smart Specialization Strategy. 24