1 Index

You’re back? Good! Welcome!!! 3 Pedro R. Lavia 3 Renato Lenzi 4

Tuesday, March 5th 6 Scientific Session #1 6 Scientific Session #2 6

Wednesday, March 6th 7 Scientific Session #3 7 Scientific Session #4 7 Scientific Session #5 8

Thursday, March 7th 9 Scientific Session #6 9

Abstracts 11 Scientific Session #1 11 Scientific Session #2 14 Scientific Session #3 17 Scientific Session #4 18 Scientific Session #5 21 Scientific Session #6 24

Posters 26 You are back? Good! Welcome!!!

1996 was a long time ago... As some of you may remember, the 24th was an amazing symposium for the EAAM and, in some aspects, a turning point in our community.

In the meantime, and after 23 years, many things changed - in our community, in our facilities, in our collections, in the perceptions of our patrons/visitors, in our personal and professional priorities... We changed and so did you (although some of you might not have been born, yet, in 1996 – and that is great). But some things do remain the same: our values, our passion, our determination, our urgencies toward Science, Education and Conservation, and the love for the animals under our care – as well as for the species and ecosystems for which they are such inspiring ambassadors. This year, we are determined to make another great EAAM Annual Symposium - because we need to continue the dream. And because we need to continue to believe in and reinforce our great family... As such, thank you for being with us and thank you for your support to our community. It is a special honour to welcome such a venerable group of professionals, who fortify, in such a distinguished way, our communal work and our common values.

On my behalf and on behalf of the Board of Mundo Aquático SA, allow me to wish you a superb exchange of experienc- es and motivations, so that our community can continue to grow, bolster its impact, while always contributing to the multi-cultural and technical complementarity that are so important to our work of promoting and increasing the wel- fare of the animals under our care!

Once again, welcome to Portugal and welcome to Zoom- arine. May you all have a magnificent stay in our beautiful Algarve and enjoy the unique country called Portugal (Lu- sitania), the land of friendship, the land of Zoomarine, and the land of the no-to-be-missed “pastel-de-nata”.

Pedro R. Lavia

Founder and President of the Board

3 Dear colleagues and friends

We are just few weeks away from our EAAM annual Symposium. The host facility staff has been working really hard to make sure this will be a memorable Symposium. The Algarve is for sure a fantastic place to meet and spend few days in a warm and welcoming environment. Our gathering is a unique and fantastic opportunity for like minded people to connect and share experience and knowledge.

The marine ecosystem is under tremendous pressure and most marine mammals are facing significant challenges in their own environment. Marine mammals in situ and ex situ research, combined with educational and conserva- tion activities are instrumental to change the current aggressive pace at which human activities and habitat chang- es are interfering with many marine species delicate status.

One of the best way to enhance the work of in situ and ex situ marine mammal experts is to insure that their knowl- edge expertise and drive is shared periodically, frequently and especially with the new generation of experts. That is why I believe the Symposium is one of the most powerful tool available to all those that have dedicated their profes- sional career to the study of marine mammals and their environment in order to insure that what we have learned, how we have learned it and why we have learned in the past is constantly passed on to the new generations of sci- entists and experts. This year will be the 47th Symposium organized by the EAAM and back in 1972, very few could have imagined how much would be shared and how many people would have been influenced because of EAAM.

I hope many of you this year will be able to make this symposium not just a memorable one but especially a great sharing and learning event. I also encourage you all to start working now to push your team mates, your organiza- tion leaders, and your colleagues that did not get involve in the symposium this year, to understand why it is ex- tremely important that they do get involve in the near future. Once again thank you to our host facility Zoomarine Portugal their staff and Mr Pedro Lavia, leader of this organization, Elio Vicente that has lead with enthusiasm and dedication the Symposium organization work.

Renato Lenzi

President-elect EAAM

4 47th Annual Symposium of the EAAM - European Association for Aquatic Mammals Zoomarine Algarve, March 4th-7th 2019

10h00 – 13h00: Institutional Member’s Meeting (closed meeting) 13h00 – 14h00: Lunch (Sponsored by Zoomarine Algarve) 14h00 – 16h00: Institutional Member’s Meeting (closed meeting) 16h30 – 19h00: EAAM Registration 19h00 – 20h30: 47th EAAM Annual Symposium Ice-breaker (Sponsored by Zoomarine Algarve)

5 Tuesday, March 5th 2019

08h30 – 09h45: Registration 09h00 – 09h10: Official Symposium Opening 09h10 – 09h30: Welcoming Speeches – Renato Lenzi, President-Elect of the EAAM & Pedro Lavia, Zoomarine’s Founder and President of the Board 09h30 – 10h30: Nachtigall, P.* (Invited Speaker) “Studying the hearing of whales and dolphins” 10h30 – 11h00: Coffee Break (Sponsored by Zoomarine Algarve)

Scientific Session #1: Chairperson – Robert Gojceta

11h00 – 11h20: Neves, J.* “Emotion, the language of conservation” 11h20 – 11h40: Delfour, F.*, Mercera, B. “Over 10 years of increasing research and conservation work in a dolphinarium housing bottlenose dolphins (Tursiops truncatus) and California sea lions (Zalophus californianus)” 11h40 – 12h00: Krasheninnikova, A.*, Tallo-Parra, O., Manteca, X., von Bayern, AMP. “Welfare and cognitive bias in captive killer whales (Orcinus orca)” 12h00 – 12h20: Clegg, ILK*, Stockin, KA “Integrating animal welfare science into conservation strategies for wild cetaceans: workshop summary from European Cetacean Society 2018 conference” 12h20 – 12h40: Gimeno-Ponce, S.*, Segura-Göthlin, S. “Killer whales (Orcinus orca) and bottlenose dolphins (Tursiops truncatus) enrichment program in “Loro Parque”” 12h40 – 13h00: Tallo-Parra, O.*, Garcia-Parraga, D., Agusti-Pujol, C., Manteca, X. “Animal welfare evaluation, improvement and research in aquaria and zoos: the Oceanogràfic-ZAWEC collaborative approach” 13h00 – 14h20: Lunch (Sponsored by Zoomarine Algarve)

Scientific Session #2: Chairperson – Katrin Baumgartner

14h20 – 15h20: Van Ferssen, L.* “Ex Situ Options for Cetacean Conservation” 15h20 – 15h40: Castro, J.*, Cid, A. “Discovering the cetaceans of southern Portugal” 15h40 – 16h00: DeRango, E.*, Schwarz, J., Trillmich, F., Krüger, O. “Does fortune favour the bold pup? Insights into multiple aspects of personality in Galápagos sea lions” 16h00 – 16h40: Coffee Break (Sponsored by Parques Reunidos) 16h40 – 17h00: Schwarz, J.*, DeRango, G., Trillmich, F., Krüger, O. “Good mothers dive deep? Foraging strategies of an endangered pinniped” 17h00 – 17h20: García-Párraga, D.*, Moore, M, Fahlman, A. “Physiologists and veterinarians working together: proposing a new paradigm on how marine mammals may minimize nitrogen uptake during diving” 17h30 – 18h30: Marine Mammal TAG meeting (closed meeting) 6 Wednesday, March 6th 2019

08h30 – 09h00: Registration

Scientific Session #3: Chairperson – Agustín López Goya

09h00 – 10h00: Matos, J.* (Invited Speaker) “Animal Molecular Genetics: It’s all about SEQs” 10h00 – 10h20: Flanagan, C.*, Lavia, P., Santos, N., Pisa, P., Camacho, G., Curado, A., Charraz, B., Neto, M. “A problem, a crazy idea, an amazing engineering solution… problem solved!” 10h20 – 10h40: Andreini, R.*, Piscopo, G., Cavedo, B., Gridelli S., Zaccaroni, A. “Dietic and toxicological parameters evaluation in Aonyx cinereus” 10h40 – 11h20: Coffee Break (Sponsored by Zoomarine Algarve)

Scientific Session #4: Chairperson – Guillermo J. Sanchez Contreras

11h20 – 11h40: Bunskoek, P.*, De Haan, K. “Pulmonary Aspergillosis in 6 bottlenose dolphins (Tursiops truncatus): culture results, sensitivity patterns and treatment results” 11h40– 12h00: Leal, M., Flanagan, C., Urbani, N.*, Silva, J., Barny, R. “Medical management of a respiratory mucormycosis infection in a bottlenose dolphin (Tursiops truncatus) calf” 12h00 – 12h20: Rubio-Garcia, A.*, John W. Rossen, J.W., Friedrich, A.W., Wagenaar, J.A., van Zeijl, J.H. “Methicillin resistant Staphylococcus aureus in a harbour seal (Phoca vitulina) with endocarditis” 12h20 – 12h40: Scala, C.*, Brasseur, I. “Diversification of the diets of marine animals at Marineland Côte d’Azur” 12h40 – 12h45: Mercera, B.* “Case report from the adult female bottlenose dolphin with the Cushing syndrome, presented at last EAAM symposium.” 12h45 – 12h50: Rubio-Guerri, C.*, Sastre, G., Piqueras, J., Sancho, R., González, C. “Implementation of a method for the detection of infectious viruses in sea water” 12h50 – 12h55: Fogaça, C.*, Flanagan, C., Urbani, N., Fernandes, R., Silva, J., Monteiro, R., Tavares, L., Duarte, A. “Astrovirus detection in faecal samples of pinnipeds in a zoological context” 12h55 – 13h00: Zaccaroni, A., Andreini, R., Hartmann, M.G. “Validation of a method for quantification of aldosterone in marine mammals” 13h00 pm – 14h20 pm: Lunch (Sponsored by Zoo de Lisboa)

7 14h20 pm – 15h40 pm: Poster Session Ecalle B.*, Mercera B., Delfour F. “What do bottlenose dolphins say and do during a cooperative task?” Kreivinienė, B.; Kleiva, Ž.; Vaišvilaitė, V.* “Welfare Aspects of Black Sea Bottlenose Dolphins and Humans during Animal Interaction Programmsh a Field for Discussion” Leal, M.*, Umarji, S., Silva, J., Flanagan, C., Urbani, N., Barny, R. “Periarticular Gold Bead Implantation and Ozone Therapy as an Integrative Approach to Chronic Elbow Joint Pain in a California Sea Lion (Zalophus californianus)” Saviano, P.*, Scala, C., Oesterwind, M., Fiorucci, L. “Preliminary Results of Normal Abdominal Ultrasonographic Imaging of Killer Whales (Orcinus orca)” Tallo-Parra, O.*, Almunia, J., Manteca, X. “Identification and study of potential welfare indicators in killer whales (Orcinus orca)” 14h20 – 15h40: Roundtable Discussion on Anaesthesia 15h00 – 15h40: Coffee Break (Sponsored by Zoomarine Italia)

Scientific Session #5: Chairperson – Birgitta Mercera

15h40 – 16h00: Serres, A.*, Hao, Y., Wang, D. “Agonistic interactions and dominance relationships in three groups of captive cetaceansh method of assessment and inter-species comparison” 16h00 – 16h20: Bigiani, S.*, Pilenga, C. “Use of the anticipatory behavior in Tursiops truncatus to distinguish different level of interest inside the same event” 16h20 – 16h40: Martinez, A.*, Martínez, J. “Voluntary stretcher: Yes or Yes!” 16h40 – 17h00: Neto, M., Rodrigues, A.*, Estanqueiro, V.*, Filhó, C., Branco, V., Gregório, R., Flanagan, C. “Secondary reinforcement, a fundamental tool for variation in water work sequences in dolphin presentationsh a myth or a reality?” 17h00 – 17h20: Ruiz, R.*, Henderson, C. “How much can you remember? Operative memory capacity training as a mental stimulation in bottlenose dolphins (Tursiops truncatus)” 17h20 – 19h00: Annual General Meeting (closed meeting)

8 Thursday, March 7th 2019

08h30 – 09h00: Registration

Scientific Session #6: Chairperson – Pietro Saviano

09h00 – 10h00: Santos, M.* (Invited Speaker) “Don’t shoot the Trainer! - From Skinner to Jobs and everything in between” 10h00 – 10h20: Miedler, S., Will, H., Baumgartner, K.* “Transthoracic cardiac ultrasound examination in Bottle nose Dolphins” 10h20 – 10h40: Zaccaroni, A.*, Andreini, R., Silvi, M., Hartmann, M.G., Flanagan, C., Vicente, E. “Can paper be better than glass? Alternative method for blood sampling in marine mammals” 10h40 – 11h20: Coffee Break (Sponsored by Zoomarine Algarve) 11h20 – 11h40: Mercera, K.*, Delfour, F., Mercera, B., Pilot-Storck, F. “Non-invasive monitoring of adrenal activity in the bottlenose dolphin (Tursiops truncatus) under human care by fecal cortisol enzyme immunoassay” 11h40 – 12h00: Sánchez-Contreras, G.J.*, Gili, C., Biancani, B., Florio, D., Pussini, N. “Hepatocellular carcinoma in a South American sea lion (Otaria flavescens)” 12h00 – 12h10: Eulogy to Jeanette Thomas (1952-2018) 12h10 – 12h15: Professional farewell to Birgitta Mercera 12h15 – 13h00: Final Address from the Board and Closure 13h00 – 14h00: Lunch (Sponsored by Zoomarine Algarve) 14h00 – 14h20: Transfer to Zoomarine 14h20 – 17h30: Visit to Zoomarine 17h30 – 17h50: Transfer to Hotel 18h40 – 19h00: Transfer to Zoomarine 19h00 – 00h00: Sponsored Gala Dinner at Zoomarine (Sponsored by Zoomarine Algarve) 00h00 – 00h20: Transfer to Hotel

9 10 Abstracts

Scientific Session #1

Emotion – the language of conservation Neves, J.1

(1) Zoomarine Algarve ([email protected]) The intrinsic value of species and their ecosystems have proven to be a language that does not appeal to many sectors of society. Thus, many conservation efforts have been ineffective in changing societal behaviors in a timely manner. The IUCN’s Commission on Education and Communication campaign “Love. Not loss.” launched in 2010 started a game-changing movement. Counting on contributions from unlikely areas such as psychology or even marketing and advertising, conservation campaigns have, since then, been gaining a new momentum and social impact. Optimistic and emotional narratives and wordings (e.g. love, care, hope, etc.) are now increasingly available and accepted as more effective communication tools for behavior change. This presentation, based on some of author’s findings along its PhD project, aims to explore some of the basic human mechanisms that can and should be used in future conservation communication strategies whose goals involve greater community involvement in a cause, campaign or behavior.

Over 10 years of increasing research and conservation work in a dolphinarium housing bottlenose dol- phins (Tursiops truncatus) and California sea lions (Zalophus californianus)

Delfour, F.1, 2, Mercera, B.1 (1) Parc Astérix ([email protected]) (2) Laboratoire Ethologie Expérimentale et Comparée

Zoos, and therefore institutions rearing marine mammals under human care, are legally obliged to conduct and to participate to scientific works and conservation programs as well as education activities. However, it is not always easy to involve staff and to make directors to accept the economic aspects of these activities. This is an example from one dolphinarium that had very little activity in research and conservation ten years ago and it is now reaching its 30th published scientific papers/chapters. Moreover, its team leads a conservation program on bottlenose dol- phins in the Caribbean sea. To find motivated students, to manage their project and of course, to encourage them to publish their work; to get trainers involved and to keep them motivated, and to convince them that scientific work can be fun and can benefit the animals under their care; and finally to collaborate and/or develop conservation pro- grams are very important, not to say essential, when you need materials to feed medias and social and/or scientific events but also to present to governmental institutions and various animal welfare committees.

11 Welfare and cognitive bias in captive killer whales (Orcinus orca)

Krasheninnikova, A.1, 2, Tallo-Parra, O.4, Manteca, X.5, von Bayern, AMP. 1,2,3 (1) Max-Planck Comparative Cognition Research Station, Loro Parque Fundación (2) Max-Planck-Institute for Ornithology ([email protected], [email protected]) (3) Department of Biology, Ludwig-Maximilians-University of Munich (4) ZAWEC (Zoo Animal Welfare Education Centre), Veterinary Faculty, Universitat Autònoma de Barcelona (Xavier. [email protected], [email protected])

Robust methods to assess nonhuman animal emotion are essential for ensuring the welfare of animals under hu- man care. A promising tool for assessing animals’ emotional states is the cognitive bias test. It measures an individ- ual’s tendency to judge ambiguous information based on its emotional state or mood. In humans, cognitive biases are known as “optimism” and “pessimism” and it has been shown that being more optimistic is correlated to better subjective well-being. The cognitive bias as a tool to evaluate animals’ emotional states has been implemented in a number of mammals and birds, and very recently for bottlenose dolphins (Tursiops truncatus) whereas generally very little research on welfare indicators has been conducted in cetaceans. In the present study, we describe for the first time the characteristics of cognitive biases in killer whales (Orcinus orca), evaluate intra-individual and inter-in- dividual variability and investigate the interrelation between their cognitive bias and other potential welfare indi- cators (e.g., behaviour, hormones). Our study aids in establishing further research directions and can be practically implemented to improve the welfare of captive killer whales, e.g. by testing the efficiency of (cognitive) enrichment programs and training protocols.

Integrating animal welfare science into conservation strategies for wild cetaceans: workshop summary from European Cetacean Society 2018 conference Clegg, ILK1, Stockin, KA2,3 (1) Animal Welfare Expertise ([email protected]) (2) Animal Welfare Science and Bioethics Centre, School of Veterinary Science (3) Coastal-Marine Research Group, School of Natural and Computational Sciences, Massey University

Animal welfare science evaluates how animals are feeling as they cope with their environment. Welfare science and conservation biology have several crossovers that have recently been emphasised in the literature, since both consider the animal’s holistic response to environment. It has been suggested that sometimes, an ‘individual animal’ approach is more valid than population-level conservation, since environmental changes impact individuals differ- ently. Research into welfare assessments for individual captive dolphins have notably increased in the past decade and could be adapted to wildlife situations. In response, a workshop called ‘Welfare in the Wild’ was held at the 32nd European Cetacean Society conference in La Spezia, Italy in April 2018. The participants were 30 international researchers, conservationists and managers. The attendees completed a pre- and post-workshop survey on animal welfare, wild cetacean conservation strategies, and the integration of the two fields. Herein, we describe social sur- veys, participative exercises, and discussions around the Vaquita (Phocoena sinus) and Maui’s dolphin (Cephalorhyn- chus hectori maui) case studies. We reveal how welfare science is currently understood by those working with and for wild cetaceans, and in what contexts this nexus of the two fields might benefit conservation strategies.

12 Killer whales (Orcinus orca) and bottlenose dolphins (Tursiops truncatus) enrichment program in “Loro Parque”

Gimeno-Ponce, S.1, Segura-Göthlin, S.2 (1) University of Las Palmas de Gran Canaria, Faculty of Veterinary ([email protected]) (2) Animal Health and Food Safery Insitute (IUSA) ([email protected]) (3) Loro Parque Fundación ([email protected])

Enrichment, “an animal husbandry principal that seeks to enhance the quality of captive animal care by identifying and providing the environmental stimuli necessary for optimal psychological and physiological wellbeing”. A variety of enrichment types was introduced into the routines of the 7 killer whales and 10 dolphins housed in “Loro Parque” (Physical, Social, Cognitive, Alimentation, Environmental). This was done to measure the effect that they have, so that they can be used appropriately to increase the welfare of the animals. A structured calendar was followed also aiming to reduce stereotypies and develop natural behaviors. The success of the different enrichment devices, activity within the group and individually was also assessed. A register system was created to collect and compare the results in behavior obtained, giving us percentages for each enrichment session, the reaction of each animal to the different devices and also the success of the devices in the whole group. The program that was used was “Excel”. Following the program, results have been obtained grading quantitatively the different enrichments (from 1, neg- ative reaction, to 5, positive reaction) allowing us to acquire information about the preferences of each animal and the general group for them.

Animal welfare evaluation, improvement and research in aquaria and zoos: the Oceanogràfic - ZAWEC col- laborative approach

Tallo-Parra, O.1, Garcia-Parraga, D.2,3, Agusti-Pujol, C.1,3, Manteca, X.1 (1) Zoo Animal Welfare Education Centre (ZAWEC), Department of Animal and Food Science, School of Veterinary Science, Universitat Autònoma de Barcelona ([email protected]) (2) Oceanográfic-Avanqua, Ciudad de las Artes y las Ciencias (3) Fundación Oceanogràfic

Animal welfare has become a top priority in zoos and aquaria. However, daily management and protection of an- imal welfare are highly complex. Among others, it involves reliable assessments of individuals’ welfare status and identification of their needs, management of strategies to improve welfare, as well as the design and application of systematic evaluations of the proposed strategies. Moreover, zoos and aquaria are excellent institutions for de- veloping wildlife welfare research. Since the beginning of 2018, the Oceanogràfic established a strong and stable collaboration with the Zoo Animal Welfare Education Centre (ZAWEC), a specialized research group from the Univer- sitat Autònoma de Barcelona with extended experience in the field of animal welfare. The collaboration included 1) staff education and training on animal welfare, 2) development of several research projects about the identification and study of welfare indicators on different species, and 3) the designation of an external and independent ZAWEC member as the Oceanogràfic’s Animal Welfare Officer. His integration into all animal welfare-related issues includes the design and implementation of 10 different types of welfare-specific records/reports; development and super- vision of the environmental enrichment program (>25 plans for welfare improvements); routine animal welfare 13 monitoring and consultancies; and participation as an external member in all welfare-related committees, among others. Benefiting from the complementary profile between aquarium and university staff, the Oceanogràfic has achieved significant improvements in theoretical and practical approaches for animal welfare assessment, man- agement and research.

Scientific Session #2

Ex Situ Options for Cetacean Conservation

Van Ferssen, L.1 (1) Zoo Nurnberg, Germany

For many animal species (mainly mammals and birds) ex situ Management proved to be the last resource in order to save species from extinction. Paired with in situ conservation the IUCN defined this integrated, science-based species conservation planning process as the ‘One Plan Approach’ (OPA). In the last decades the population of many wild animal species could be secured thanks to the application of this integrated conservation approach. Regarding cetaceans, the few attempts to safe the Baiji and the Vaquita by using ex situ management had to be suspended and unveiled various information gaps that have to be resolved before applying ex situ strategies to help other small cetacean species in the near future. However, the integrated approach of in situ and ex situ actions to save the Yangtze finless porpoise that started 25 years ago is an encouraging example of the ‘One Plan Approach’ for a population of small cetaceans. As many small cetaceans are facing a drastic population decline and the threats that affect these species will not decrease in a foreseeable future, the urgency to prepare a more complete set of conservation tools is evident. With that in mind in December 2018 a Workshop entitled EX SITU OPTIONS FOR CETACEAN CONSERVATION (ESOCC) was organized in Nuremberg. The workshop was designed to help to improve our understanding of all the potential options available and to ensure that all necessary tools could be made ready for use when the next small cetacean extinction crisis arise. Workshop activities included attempts to identify and elaborate lessons learned from the ef- forts to save the baiji, the Yangtze finless porpoise, and the vaquita; reviews of the conservation status of the other representative small cetacean species or populations of concern; and discussion of the ex situ options available that could be beneficial in helping to prevent extinction.

Discovering the cetaceans of southern Portugal

Castro, J.1,2, Cid, A.1 (1) AIMM – Associação para a Investigação do Meio Marinho ([email protected]) (2) MARE – Marine and Environmental Sciences Centre, Laboratório Marítimo da Guia ([email protected])

The Algarve, located in the south of Portugal is well known for its long coastline facing the rich Atlantic Ocean. Due to the large amount of tourism in the area, over the past two decades the whale watching industry has expanded very rapidly. However, although it is the area of mainland Portugal with the highest concentration of whale watch- ing activities, scientifically, very little is known about the megafauna inhabiting its waters. To address this, for the past 10 years a long-term study has been conducted to assess the biodiversity and abundance of cetaceans in 14 the area. As a part of this research, dedicated scientific surveys were conducted and data from commercial whale watching trips was gathered, resulting in the identification of eleven species of cetaceans, both odontocetes and mysticetes, the former accounting for 97% of the total number of sightings. This region is characterized by an in- tense wind-driven upwelling season from March to October, creating the ideal conditions for the occurrence of dolphins. Data indicates that this could be an important breeding ground for some species, particularly for short- beaked common dolphin, and photo-id data shows that common bottlenose dolphin, Risso’s dolphin and killer whales also present a certain site fidelity.

Does fortune favour the bold pup? Insights into multiple aspects of personality in Galápagos sea lions

DeRango, E.1, Schwarz, J.1 , Trillmich, F.1 , Krüger, O.1 (1) Bielefeld University, Department of Animal Behaviour ([email protected])

Animal personalities and their relative degree of behavioral plasticity have been demonstrated to be adaptive and associated with ecologically important traits. While the concept is well established in a broad range of species, studies with free-ranging pinnipeds remain relatively rare. This presentation will provide an overview of animal personality utilizing long-term data with a presently declining colony of Galápagos sea lions. This species resides in a habitat characterized by environmental fluctuation and perturbations, which may select for differences in person- ality and plasticity within individuals. We developed and integrate several behavioral assays and ethograms, such as responses to novel objects, capture and human approach, to determine whether consistent individual differences exist within a cohort of sea lion pups. Our results suggest correlations between boldness and exploration, and these traits remained repeatable and stable within the first year of life. Maternal age and body condition also affected variation in pup’s responses to these tests which suggests that fitness-relevant characteristics of mothers may influ- ence pup personality. We propose that animal personality measured during early ontogeny in this species may be relevant for coping strategies during later life history events within a changing environment.

Good mothers dive deep? Foraging strategies of an endangered pinniped

Schwarz, J.1, DeRango, G.1 , Trillmich, F.1 , Krüger, O.1 (1) Bielefeld University, Department of Animal Behaviour ([email protected])

Hunting in the tropical waters off the coast of Ecuador, Galápagos sea lions are confronted with strong, irregular fluctuations in prey abundance due to El Niño and La Niña events. Despite living in dynamic environments, marine predators frequently exhibit different, stable individual foraging strategies. Different foraging strategies are espe- cially close connected to the survival of an individual or its offspring, with large influences on population dynamics. Hence we are studying individual differences in foraging behavior of Galapagos sea lion mothers during current El Niño conditions. Using biologgers, we collected location, dive depth and acceleration data and related it to the age and physiology of 19 mother offspring pairs. We expect to find 1) individual variation in dive behavior, such as bout

15 duration, volume of 3D utilization distribution and depth of prey chasing events, and 2) an influence of those strat- egies on maternal body condition and on the weight gain of their pups, two important components of individual fitness. Studying the different diving strategies of this decreasing population of pinnipeds can not only give us an insight into how these animals handle current and future challenges, but also help us gain a better understanding of how these strategies develop. Physiologists and veterinarians working together: proposing a new paradigm on how marine mammals may minimize nitrogen uptake during diving

García-Párraga, D.1, Moore, M.2 , Fahlman, A.1, 2 (1) Fundación Oceanografic ([email protected]) (2) Woods Hole Oceanographic Institution

Studies of beached and bycaught cetaceans and sea turtles imply that breath-hold divers may, under unusual circumstances, develop gas emboli (GE) that result in decompression sickness symptoms/lesions (DCS). Passive lung compression at depth has been considered the main theoretical basis for limiting N2 uptake and avoiding GE. Different models suggest that the depth for complete alveolar collapse may be much greater than original- ly estimated. In fact, theoretical modeling of tissue and blood gas dynamics of breath-hold divers indicates that our current understanding of diving physiology is incomplete as the models predict blood and tissue N2 tensions suggestive of a high probability of DCS following most natural dive profiles. We combined results from studies on anatomy, imaging and physiology in different animal species and humans to develop a novel hypothesis on how breath hold divers may have evolved volitional control of gas exchange at depth. We hypothesize that cetaceans may have developed an active mechanism to create a functional shunt through management of a ventilation per- fusion mismatch in the lung. Man-made disturbances, causing stress at depth, could alter ventilation/perfusion mismatch increasing the risk for GE.

16 Scientific Session #3

A problem, a crazy idea, an amazing engineering solution… problem solved!

Flanagan, C.1, Lavia, P.1 , Santos, N.1 , Pisa, P.1 , Camacho, G.1 , Curado, A.1 , Charraz, B.1 , Neto, M.1 (1) Zoomarine Algarve ([email protected])

Zoomarine has two main Lagoon enclosures (the Blue Lagoon and the Tropical Enseada). The Blue Lagoon was built first and has 2 lagoons and a total of 2300 m3 of water. Several years later and in order to accommodate a growing collection the “Tropical Enseada” was built which has four lagoons and a large maternity pool that holds a total of 4640 m3 of water. These two Lagoon areas are situated next to each other but 16 meters apart and were built on different levels, as they are located on a hill and there is a 3,8 meter elevation difference between them. Having such physically separated enclosures, meant every time we decided to move one of our dolphins (for so- cial, medical or reproduction reasons) to a different enclosure we had to do so using a stretcher, trainers physical strength and a truck with all the risks that even a very short distance transport can entail. Then one of our team members came up with an idea to build our very own “Panama channel” in between the two Lagoon enclosures. Using a connecting pool with a medical platform and water tight gates it will allow us to transfer our dolphins from one enclosure to another as easily as from one pool to the next. The construction is currently almost finished. We would like to share the project, the planning and the complications we faced developing this new concept. It just goes to show that sometimes the solution to a problem may just be that crazy idea.

Dietic and toxicological parameters evaluation in Aonyx cinereus

Andreini, R.1, Piscopo, G.2, Cavedo, B.2, Gridelli S.3, Zaccaroni, A.1 (1) Department Veterinary Medical Sciences, University of Bologna ([email protected]) (2) Zoo delle Maitine (3) Costa Edutainment S.p.A., Acquario di Cattolica-Piazzale delle Nazioni

Aonyx cinereus is the smallest otter existing, listed as Vulnerable under IUCN red list. Data on diet and nutritional intake of this species in the wild are scarce. Despite this, one of the most important parameters in the management of Aonyx cinereus in captivity, is the development of an appropriate diet program. Present study aims at comparing two different groups of A. cinereus, one hosted at Acquario Le Navi of Cattolica and one at Maitine Zoo in Beneven- to. We collected samples of food from each feed batch once, and faces of otters on weekly basis for several weeks, evaluating protein, lipidic and caloric content of the daily food rations of the animals. Moreover, we analysed con- centration of trace elements in food and faeces, to define if the diet is balanced and safe and to acquire information about the metabolic processes of clawed otters. This study compares the dietary composition and trace elements exposure in two different otter groups; it also provides scientific data on Aonyx cinereus nutritional needs, metab- olism and possible exposure to detrimental contaminants, giving useful information to implement the welfare and the health status of clawed otters under human care.

17 Scientific Session #4

Pulmonary Aspergillosis in 6 bottlenose dolphins (Tursiops truncatus): culture results, sensitivity patterns and treatment results

Bunskoek, P.1, De Haan, K.1 (1) Dolfinarium ([email protected])

During 5 years 6 bottlenose dolphins living in an outdoor lagoon with filtrated natural seawater have developed pulmonary Aspergillosis. In all 6 animals several bronchoscopies were performed to diagnose and monitor the problem and to take samples for fungal cultures. The samples were sent to either the microbiology laboratory of the local hospital or to a more specialized mycology laboratory. In 43 samples Aspergillus fumigatus was cultured. In all but one sample sensitivity was tested for at least itraconazole, voriconazole and posaconazole. Not in all sub- mitted samples A. fumigatus was cultured. This could be due to sampling method or treatment with antifungals. Treatments were chosen based on sensitivity patterns. Resistance against all azoles was high looking at human minimum inhibitory concentrations (MICs). Resistance against itraconazole was 38.1 %, against voriconazole 60.5 % and posaconazole 59.5 %. As dolphins can reach higher plasmalevels of posaconazole than humans MICs of 0.5 and 1 mg/L could be classified as intermediate sensitivity in these animals. Taking this into account the resistance against posaconazole would be 0 %. In most cases animals respond to treatment with posaconazole alone. Recently a combination therapy with terbinafine was used with promising results and without severe side effects.

Medical management of a respiratory mucormycosis infection in a bottlenose dolphin (Tursiops truncatus) calf

Leal, M.1, Flanagan, C.1 , Urbani, N.1 , Silva, J.1 , Barny, R.1 (1) Zoomarine Algarve ([email protected])

An 11-month-old male bottlenose dolphin calf presented with a persistent leukocytosis with no clinical signs. During a routine fecal examination as part of a monthly check-up, fungal structures were found. After molecular identification, PCR and sequencing, Cunninghamella bertholletiae was identified – an etiologic agent of mucormy- cosis. Rare episodes of cough were later reported, along with absence of weight gain, being these two the only clinical signs expressed. Radiographic examination of the thorax (VD and LL) showed the presence of a bronchoalveolar pattern on both the right and left lung apex. A bronchoscopy was performed (subtle sedation with midazolam 0,025mg/kg IV), which revealed multiple whitish lesions, diffusely distributed on the tracheal and bronchial submucosa, images were com- patible with a severe fungal infection. Medical treatment prescribed was posoconazole (5mg/kg PO BID), aerosolized liposomal amphotericin B (BID), bromhexine (8 mg PO BID), vitamin C (1000mg PO BID), probiotics and ozone therapy (rectal/oral). Treatment fol- low-up was conducted with hematology and blood biochemistry, fecal fungal culture and bronchoscopies.

18 Methicillin resistant Staphylococcus aureus in a harbour seal (Phoca vitulina) with endocarditis

Rubio-Garcia, A.1, John W. Rossen, J.W.2, Friedrich, A.W.2 , Wagenaar, J.A.23, van Zeijl, J.H.4 (1) Sealcentre Pieterburen ([email protected]) (2) Department of Medical Microbiology and Infection Prevention, University of Groningen (3) Department of Infectious Diseases and Immunology, Faculty of Veterinary Medicine, Utrecht University (4) Department of Medical Microbiology, Izore Center for Infectious Diseases

A 5-month-old male harbour seal (Phoca vitulina) was admitted for rehabilitation to the Sealcentre Pieterburen on November 16th, 2015. During the initial veterinary examination parasitic pneumonia and secondary bacterial pneumonia were suspected. Therefore, the seal received antiparasitic and antibiotic treatment and appeared to be recovering but died unexpectedly after several weeks. The main necropsy findings were a large amount of blood in the pericardial space and a perforating orifice in the aortic wall. Histopathological examination of the aorta revealed mural necrosis with haemorrhage, suppurative to mixed inflammation and presence of granular cystic structures. Bacterial culture resulted in the isolation of a methicillin resistant Staphylococcus aureus (MRSA) from the pericardial effusion. Subsequent culture of rectal swabs collected at arrival and during the seal rehabilitation process revealed that the animal was already colonized when admitted to the Sealcentre. MRSA has been isolated from marine mam- mals before, including from two harbour seals, two pilot whales (Globicephala macrorhynchus) and five bottlenose dolphins (Tursiops truncatus). In addition, Staphylococus aureus has been related to endocarditis in a harp seal. How- ever, to our knowledge this is the first report of MRSA associated endocarditis in seals.

Diversification of the diets of marine animals at Marineland Côte d’Azur

Scala, C.1, Brasseur, I.1 (1) Marineland Côte d’Azur ([email protected])

Provide adequate food, in terms of quantity and quality, is an important part of good husbandry and well-being of zoo animals. Inadequate diets can lead to nutritional diseases well described in piscivorous species. Most species of marine animals eat a large diversity of species in the wild; but prey species diversity is lower under managed care. In an effort to continuously improve the health and the conditions of maintenance of animals at Marineland Côte d’Azur, the diets have been revised. Many factors were taken into account when establishing the new diets. The most important is that the choice of the food items is based on the main prey consumed in the wild, the energetic value and the nutritional composition of the prey species. The other points of consideration were the price of the prey species, their availability on the markets, the season of fishing, and the maximum shelf life acceptable to main- tain a good nutritional composition. Food quantities were determined using metabolic rate estimation equations and other factors like the season and the physiologic status. But in fine, food quantities were adjusted on a daily basis depending on the animal behavior, as all animals are fed nearly ad libitum in the park.

Case report from the adult female bottlenose dolphin with the Cushing syndrome, presented at last EAAM symposium Mercera, B.1 (1) Parc Asterix ([email protected]) 19 Femke, 37 years old female bottlenose dolphin (Tursiops truncatus) has been diagnosed having the Cushing syn- drome since two years now. Her therapy development and large improvements in behaviors and appetite; her “weird-looking” body condition with fat disposes and muscle loss and her frequent surface resting behavior during the day; and finally, animal right groups using her case as their battlefield and how trainers face the criticism will be presented.

Implementation of a method for the detection of infectious viruses in sea water

Rubio-Guerri, C.1, Sastre, G.2, Piqueras, J.2 , Sancho, R.2 , González, C.2 (1) Fundación Oceanografic ([email protected]) (2) GAMASER

Several studies confirm the abundance of viruses in the oceans, establishing an average around 107 viruses/ml. Some of these viruses have produced thousands of deaths in dolphins as cetacean morbillivirus. In addition, other non-lethal viruses such as Herpesvirus has been shown to produce skin lesions that cause great problems in con- trolled environment marine mammals. That is why it would be interesting to know the relationship that exists be- tween the viruses in the water and the viruses found in the animals. Therefore, Oceanografic has been the model to adapt the molecular diagnostic protocols used in marine mammals to sea water. In the selected tanks, viral particles in 30 l was concentrated using a fiber ultrafiltration system equipped with a disposable dialysis filter (30 kDa mo- lecular weight cut) immediately after sample collection. The viral particles contained in the resulting concentrates will be concentrated and purified to subsequently perform the DNA and RNA extraction, and conventional PCRs were used to identify different viruses (Morbillivirus, Herpesvirus, Adenovirus, Poxvirus, Papillomavirus). This study will allow us to know more about the epidemiology of these viruses and the possibility of finding human viruses in these controlled environments and how affect this relationship between humans and animals.

Astrovirus detection in faecal samples of pinnipeds in a zoological context

Fogaça, C.1,3, Flanagan, C.2, Urbani, N.2 , Fernandes, R.2 , Silva, J.2 , Monteiro, R.2 , Tavares, L.3, Duarte, A.3 (1) Centro de Investigação Interdisciplinar em Sanidade Animal (CIISA), Faculdade de Medicina Veterinária, Univer- sidade de Lisboa (2) Zoomarine Algarve (3) CIISA, Faculdade de Medicina Veterinária ([email protected])

Astrovirus are viruses with high genetic diversity, infect a wide range of hosts and are considered one of the most important causes of pediatric acute gastroenteritis. In order to collect information regarding Astrovirus in pinnipeds, faecal samples from two zoological collections, Zoomarine and Vasco da Gama Aquarium, were collected and a molecular survey was conducted by conventional PCR. Pan-astrovirus primers targeting the conserved regions of RdRp (viral polymerase), encoded by the ORF1b domain were used with the amplification conditions previously described by Chu et al. 2008. From a total of 19 pinnipeds of 4 different species, 3 positive samples were identified (15.7%), namely 1 South Afri- can fur seal (Arctocephalus pusillus pusillus), 1 Californian sea lion (Zalophus californianus) and 1 harbor seal (Phoca vitulina). The phylogenetic analysis of 41 amino-acidic sequences revealed an independent cluster, included the three Astro- 20 virus Portuguese sequences. This work presents the first report of astrovirus detection on faecal samples from harbour seal (P.vitulina) and South African fur seal (A.p.pusillus) species worldwide and in a zoological context, improving the existing knowledge con- cerning astrovirus host range. It also prompted to future epidemiological studies regarding viral pathogens in ma- rine mammal populations under human care.

Validation of a method for quantification of aldosterone in marine mammals

Zaccaroni, A.1,2, Andreini, R.1, Hartmann, M.G.2 (1) Department Veterinary Medical Sciences, University of Bologna ([email protected]) (2) MarLab

The standard procedure for monitoring stress in marine mammals is the determination of cortisol. Anyway, since more than 40 years, aldosterone has been proposed as more reliable marker of external, chronic stress with respect to cortisol, whose concentrations can be modified also by internal stressors (i.e., illness, physiological alterations). Despite this, aldosterone testing in marine mammals is little considered and used and all recent publications focus on cortisol as the exclusive stress marker, so at present the test has not been validated in these species. Present work reports about the validation of an ELISA test for aldosterone in different marine mammal species, and some application to real case situations. Plasma collected from bottlenose dolphins, orcas, Risso’s dolphin, sea lions, seals and polar bear have been analyzed for aldosterone, as well as serial diluted (1:2, 1:4 and 1:8) samples from the same species. The analysis of diluted samples proved for a good parallelism of analytical data in all species, so the test could be considered as reliable and repeatable. Data from a real case confirm that aldosterone is a good marker of chronic stress and is less sensitive to those internal or acute factors which dramatically impact on cortisol.

Scientific Session #5

Agonistic interactions and dominance relationships in three groups of captive cetaceans: method of as- sessment and inter-species comparison

Serres, A.1, Hao, Y., Wang, D. (1) Institute of Hydrobiology, Chinese Academy of Sciences, China ([email protected])

Social group dynamics are known to impact cetaceans’ health and welfare in captivity. Social dominance and ago- nistic interactions that occur to establish and maintain these relationships might express themselves differently be- tween species and groups, and could be used to monitor social changes. This is the first comparative and quantita- tive study of agonistic interactions and dominance relationships among three groups of captive cetaceans (Yangtze finless porpoises, East Asian finless porpoises, and bottlenose dolphins) and the first use of the Elo-rating method to assess dominance in cetaceans groups. This work also describes agonistic behaviours in finless porpoises for the first time. We report that agonistic behaviours differed among groups and dolphins’ agonistic encounters lasted significantly longer than porpoises’. Animals engaged more often in agonistic interactions when an unusual social event occurred (e.g. reunion of social groups or introduction of new animals). Finally, there was no influence of sex in the dominance hierarchy of porpoises whereas male dolphins were dominant over females. We recommend daily monitoring of agonistic behaviour and social dominance in captive cetaceans groups using similar methods 21 for detecting social changes early on that might develop to threaten the animals’ physical and psychological health and welfare.

Use of the anticipatory behavior in Tursiops truncatus to distinguish different level of interest inside the same event

Bigiani, S.1, Pilenga, C.2 (1) University of Pisa, Italy ([email protected]) (2) Zoomarine, Italy

Anticipatory behavior describes the actions taken to prepare for an upcoming event. In this study we tested the anticipatory behavior to distinguish different levels of interest inside the same event. We started by measuring the interaction time of seven bottlenose dolphins (Tursiops truncatus) with twenty game objects, then we took the objects with the highest level of interest (HLI) and those with the lowest level (LLI). In this way the event: interaction with enrichment objects, has been divided in two levels of interest according to which objects we gave to the dolphins. We trained dolphins to associate the arrive of different objects with different visual signal, then we measured how much time each dolphin showed anticipatory behaviors during the five minutes from the beginning of the signal to the arrive of the objects. Analyzing this data, using a general linear model, we found that anticipatory behavior was five time higher before the arrive of objects with HLI’s than those with LLI. This means that anticipatory behavior has been able to discriminate the two levels of interest inside the event. To conclude the study, we also tried to correlate this result with the dolphins’ spatial behavior.

Voluntary stretcher: Yes or Yes!

Martinez, A.1, Martínez, J.1 (1) Selwo Marina, Spain ([email protected])

A four month dolphin facility improvement meant that our dolphins at Selwo Marina were relocated to a different facility. The animals were housed in a huge pool with no holding pools, and therefore there was no possibility of gating him to a separate lagoon, or the use of a medical pool to help with separating him from the others. During the time here, it was announced that one of the dolphins, Rocco, would be transferred to Zoomarine Portu- gal earlier than the date determined for the other dolphins to move back to Selwo. It was decided that a voluntary stretcher must be trained, yes or yes. This would eliminate the stress of draining the pool, using nets, or undergoing invasive captures. Due to unforeseen circumstances, Rocco’s transport to Portugal was delayed. He was moved back to Selwo Marina in March 2018 along with the other dolphins. The behaviour was completed at the new facility for the next transport. The team wish to share their experience with you of training a complex behaviour in challenging conditions and working with difficult circumstances. This is to show facilities that through training it is possible to achieve goals that promote animal welfare.

22 Secondary reinforcement, a fundamental tool for variation in water work sequences in dolphin presenta- tion: a myth or a reality?

Neto, M.1, Rodrigues, A., Estanqueiro, V., Filhó, C., Branco, V., Gregório, R., Flanagan, C. (1) Zoomarine Albufeira, Portugal ([email protected])

Secondary reinforcement is an extremely important tool in any good training program. However, during water sequences, it seems not to always be used consistently. Trainers tend to only use primary reinforcement, believing that it is the only way they will be able to maintain the overall quality of the behavior, avoid losing certain behaviors and creating others like aggression in certain situations (e.g. having trainers in the water with a great number of dolphins). Zoomarine’s dolphinarium houses a total of 8 bottlenose dolphins (Tursiops truncatus), 2 males and 6 females, with ages that range from 18 months to 40 years old that participate in our public presentation. Our presentation has two main objectives, to promote our educational and conservation message and perform a set of aquatic sequenc- es where we show the audience the trusting, caring and respectful relationship that exists between these magnifi- cent ocean ambassadors and the team of trainers. Our goal will always be to show the love, tenderness and respect we have for these animals through the relationships our public sees in our presentation. Experienced trainers recognize the importance of secondary reinforcement, not only for the variety of the reinforce- ment itself but also for increasing motivation. In this presentation we want to share our experience in training water sequences with our dolphins with the consistent use of secondary reinforcement.

How much can you remember? Operative memory capacity training as a mental stimulation in bottlenose dolphins (Tursiops truncatus)

Ruiz, R.1, Henderson, C.2 (1) WHIT Welfare and Husbandry Innovative Training ([email protected]) (2) Adaland dolphin park, Kusadsy, Turkey

As a continuation of the presentation we did last year in Rome, Italy, in this paper we describe the details of the adaptation of a dolphin’s memory during discriminative memory training. To examine the operational memory and the retention capacity, we trained our dolphin Maya to memorize boards varying in shapes and colors. In this training, the dolphin had to retain certain information while performing anoth- er task. The training program consisted of two stages: 1) After presenting different boards, the dolphin could retain in memory 17 shapes. In the same session, the dolphin had to a) remember and choose a single shape according to the corresponding SD, for up to ten different shapes; (b) remember two arrows indicating which direction to go (left or right); and (c) remember five shapes as an SD to perform five different behaviors. 2) The dolphin had to retain on memory all this information until a future training session after 12 and up to 48 hours later. The stimulation of the operational memory using visual and abstract stimuli is very useful for the mental stimula- tion of our animals. The results of this training program confirm great discrimination and retention capacities of the dolphin Maya. 23 Scientific Session #6

Transthoracic cardiac ultrasound examination in Bottlenose Dolphins

Miedler, S.1, Will, H.2, Baumgartner, K.2 (1) Cardiology Valencia, Spain (2) Zoo Nuremberg, Germany

Transthoracic cardiac ultrasound examination in Bottlenose Dolphins was first established at the Nuremberg Zoo in 2001 and has become a valuable diagnostic tool in recent years. The following two cases will be described: Moby (57 years): In this old male an anamnesis of chronic kidney disease, which can lead to systemic hypertension and increased preload, initiated an additional examination in October 2017. The results could be compared to those from a previous examination in 2011. Several alterations, possibly caused by the kidney disease, were detected (e.g. focal hyperechogenic spots of the myocardium, mild concentric hypertrophy of the left ventricle, signs of diastolic dysfunction, regurgitation at the mitral valve, mild to moderate left atrial volume overload and a tricuspid valve regurgitation). Rocco (13 years): Due to a heart murmur in his medical history, a cardiac examination was performed prior to a planned transport. The absence of cardiac abnormalities led to the assumption that the formerly diagnosed cardiac murmur could have been a consequence of temporary changes in blood viscosity due to a disease or an anemic status, which can cause temporary valve insufficiencies. Both cases proved the benefit of the transthoracic cardiac ultrasound examination as a supporting diagnostic tool.

Can paper be better than glass? Alternative method for blood sampling in marine mammals

Zaccaroni, A.1,2, Andreini, R.1, Silvi, M.1, Hartmann, M.G.2, Flanagan, C.3, Vicente, E.3 (1) Department Veterinary Medical Sciences, University of Bologna ([email protected]) (2) MarLab, France (3) Zoomarine Algarve, Portugal

One main issue in the analysis of trace elements and of hormones in marine mammals is shipping samples to exter- nal laboratories. Samples should be shipped and maintained cooled, and the fastest shipping should be warranted. When working in the field or in parks located in areas with reduced transport connection, this can be hard to do, if not impossible. An alternative way to store and ship samples is the use of dried blood drops on paper. We tested the feasibility of using of paper protein saver cards for the collection and the analysis of trace elements and hormones in marine mammals. Blood and plasma were voluntary collected from dolphins and pinnipeds, and contemporarily blood drops (100 µl) were collected on cards. Sample were analyzed for trace elements and hormones. Metals were analyzed in whole blood and in one drop of blood on card, and hormones were quantified in plasma and in blood drop (in double) after extraction from paper with ethanol. In both cases, paper was analyzed as blank. Obtained data were compared to verify if a good correlation between results exists and to prove if paper cards can be a good alternative to “standard” collection methods.

24 Non-invasive monitoring of adrenal activity in the bottlenose dolphin (Tursiops truncatus) under human care by fecal cortisol enzyme immunoassay

Mercera, K.1, Delfour, F.2,3, Mercera, B.3, Pilot-Storck, F.4 (1) École nationale vétérinaire d’Alfort, France ([email protected]) (2) Laboratoire d’Éthologie Expérimentale et Comparée, Université Paris (3) Parc Astérix, France (4) Unité pédagogique de physiologie, éthologie et génétique, France

This study is part of the integrative approach assessing captive bottlenose dolphins (Tursiops truncatus) welfare by looking at a potential indicator of stress, which is the fecal cortisol concentration that increases when the hypotha- lamic-pituitary-adrenal axis (HPA) is activated. Our objectives were to set up a reliable protocol to measure out fecal cortisol by enzyme-immuno assay (EIA) and to assess fecal cortisol variation over time. Fecal samples were voluntarily and non-invasively taken on 15 bottlenose dolphins housed in four european dolph- inaria during three collection periods of five weeks (september 2017; november-december 2017; april 2018). Sam- ples were collected between four to ten times for each dolphin during each period. We validated this EIA and dis- criminated conditions of stimulation (pregnancy) and inhibition (megestrol acetate administration) of the HPA. The results show that males had significantly higher fecal cortisol concentrations than non-pregnant females, and fecal cortisol was significantly higher in pregnant females than in all other dolphins. Moreover, males, but not females, appeared to have higher concentrations during spring than during autumn and winter. To conclude, fecal cortisol EIA may help monitoring bottlenose dolphins’ adrenal activity. However, we discuss the strict relevance of a fecal cortisol increase when assessing dolphins’ level of stress.

Hepatocellular carcinoma in a South American sea lion (Otaria flavescens)

Sánchez-Contreras, G.J.1, Gili, C.2, Biancani, B.3, Florio, D.4, Pussini, N.2 (1) Mediterraneo Marine Park, Malta ([email protected]) (2) Acquario di Genova, Italy (3) Oltremare, Italy (4) Dept. Veterinary Medical Sciences, University of Bologna

A male South American sea lion (Otaria flavescens) presented on December 2017 with intermittent anorexia. Phys- ical check-up was performed and blood collected for CBC showed mild neutrophilic leucocytosis. Long lasting an- tibiotic was administered. Follow up blood checks showed severe neutrophilic leucocytosis and severe increase of liver parameters (up to ALT 498 UI/L, AST 501.7 UI/L, GGT 1152 UI/L, total Bilirrubin 9.45 mg/dL). In the meantime, the animal continued having intermittent anorexia and lost weight. The animal was then force fed to provide fluids, nutritional support and therapies. Tests were performed to exclude TB. Ultrasound examination revealed dyschro- mia, enlargement and reduced irrigation of the liver. Strong hypoechogenicity of the stomach and ascites was ob- served. Hepatic protectors were added to the therapy together with oral antibiotherapy and anti-inflammatories. Despite all the efforts the animal lost over 40 kilos of body weight, unresponsive to any therapy and in the last days lethargic and almost comatose. Thus, it was decided to proceed with human euthanasia. Necropsy was immediately performed and major lesions were observed in the liver, confirming the US examination. Histopathological exam showed the presence of a hepatocellular carcinoma with evidence of metastasis in the lungs. 25 Poster session

What do bottlenose dolphins say and do during a cooperative task?

Ecalle B.1, Mercera B.1, Delfour F.1,2 (1) Parc Astérix, France ([email protected]) (2) Laboratoire Ethologie Expérimentale et Comparée, University of Paris Cooperation occurs when two or more individuals act together to achieve a common goal and in order to ad- just to each other’s behaviour, partners exchange information. Bottlenose dolphins (Tursiops truncatus) cooperate during foraging, mate acquisition, aggression or play with an increase in their sound production. However their vocalisations’ exact role remains unknown. In this study, an etho-acoustic analysis was conducted on eight bot- tlenose dolphins at Parc Astérix dolphinarium during a cooperative task using a 360° recording device. Following a desensitization phase, where three dyads were trained, dolphins were expected to cooperate by acting on the task device and to communicate to coordinate their behaviours. The results showed that dolphins solved the task in an unexpected way. First, females tended to associate together revealing the impact of the bottlenose dolphins’ social structure on this task. The displayed “investigation” behaviour might have served to gain information and to plan future actions. Finally, burst-pulsed sounds, more produced during social behaviours than during solitary behaviours, seemed to have a communicative function. We discuss the absence of competition and the possible display of play behaviours; dolphins could have play with the device, acting on their own in a sequential way that led to its opening.

Welfare Aspects of Black Sea Bottlenose Dolphins and Humans during Animal Interaction Programs: a Field for Discussion

Kreivinienė, B.; Kleiva, Ž.; Vaišvilaitė, V.1 (1) Lithuanian Sea Museum ([email protected])

Lithuanian Sea Museum together with Kurtuvenai Regional Park Directorate has launched a scientific health-bio- technology project co-financed by Structural Funds of the European Union (Project Nr. 01.2.1-LVPA-V-835-01-0008) with title: “Developing the Methodology of Animal Assisted Therapy Based on Scientific Research and Integrating It into the Concept of Holistic Medicine”. The main goal of the project is to systemize the existing scientific knowl- edge and practices in order to develop a concept of both-sided welfare aspects of humans and animals during intervention programs. Project will consist of three stages: in the first stage, systemic meta-analysis and conceptual framework of welfare evaluation will be created. In the second stage, a prototype of therapeutic effects on humans and animals will be designed basing on the research results. The third stage will cover collaborative work with na- tional institutions and ministries. Project is covering three-kinds of animal assisted intervention programs: dolphin assisted, hippo, and canine therapy. Three multidisciplinary research groups collaborating with foreign scientists are working on this project: Vilnius University, Klaipeda University, and Public Enterprise “Vaiko raida”. Preliminary results will be presented in the poster.

26 Periarticular Gold Bead Implantation and Ozone Therapy as an Integrative Approach to Chronic Elbow Joint Pain in a California Sea Lion (Zalophus californianus)

Leal, M.1, Umarji, S.2, Silva, J., Flanagan, C., Urbani, N., Barny, R. (1) Zoomarine Albufeira, Portugal ([email protected]) (2) ZenVet, Portugal

A neutered 130kg male, 20-year-old California sea lion (Zalophus californianus) had shown signs of pain in his left upper limb for the last 17 years. Acute pain management with NSAIDs (meloxicam or carprofen) and joint support prescription brought no significant improvement. Habitat modifications, weight control and Gabapentin (5mg BID PO for 6 days and then increasing to 10mg BID PO) were introduced as part of the rehabilitation plan. The animal improved significantly, allowing us to perform radio- graphs voluntarily, which revealed severe joint disease in his left elbow (degenerative joint disease). In order to decrease the inflammatory factors and restrict the degenerative process, we performed an innovative technique: a periarticular gold bead implantation in the affected joint, associated with ozone therapy during gen- eral anesthesia. Post-surgical examination revealed an increment of the joint function (less lameness and better posture) and ab- sence of inflammatory signs, which could be explained by the possible pain relief. The combined use of ozone therapy and periarticular gold implants as an alternative treatment in this case resulted in a successful treatment of pain, avoiding more invasive techniques (surgical correction by arthroscopy/arthroto- my) and a harder post-surgical recovery. As far as the authors are aware, this is the first report of ozone therapy and gold bead implantation in chronic joint disease management in California sea lions.

Preliminary Results of Normal Abdominal Ultrasonographic Imaging of Killer Whales (Orcinus orca)

Saviano, P.1, Scala, C.2, Oesterwind, M.2, Fiorucci, L.3 (1) Ambulatorio Veterinario Saviano Larocca, Italy ([email protected]) (2) Marineland Côte d’Azur, France (3) Division of Animal Histology and Pathology, University of Las Palmas de Gran Canaria Ultrasonographic examination is extensively described in bottlenose dolphins (Tursiops spp.), but information in killer whales (Orcinus orca) is scarce. The aim of this work is to shareour knowledge in the ultrasonographic exam- ination of the main abdominal organs of killer whales. All the sonograms were collected during routine voluntary training on four apparently healthy killer whales (4 to 19 years old) under professional care, housed at Marineland Park Côte d’Azur, France. Ultrasonography was recorded using a Logiq V2, General Electric, with a 2-5MHz curvilinear transducer. Right and left lateral recumbencies of the subjects have been used in the examination. Only abdominal organs were described. The Forestomach’s wall has been visualized thin with more evident plications than in the dolphin. The fundic chamber of killer whales, unlike dolphins, had a wall similar to that of the forestomach but with shorter plications and thinner. The pyloric chamber had an elongated shape with a linear and homogeneous wall. The hepatic margins were sharp, and the hepatic pa- renchyma had a low echogenicity, compared with dolphins. The kidneys had a classical pluri-lobulated architecture. Nowadays, organs like adrenal glands and pancreas are difficult to be accurately evaluated. 27 Identification and study of potential welfare indicators in killer whales (Orcinus orca)

Tallo-Parra, O.1, Almunia, J.2, Manteca, X.1 (1) Zoo Animal Welfare Education Centre (ZAWEC), Universitat Autònoma de Barcelona ([email protected]) (2) Loro Parque Fundación, Spain

There is a growing interest and need for zoos to develop and implement welfare indicators that are practical to use and provide meaningful results that help management decisions. Furthermore, zoo animals can be used as a model for developing welfare indicators that would also be useful for studying the welfare of their free-ranging conspe- cifics. The present study aimed to identify and study welfare indicators in killer whales (Orcinus orca). For this pur- pose, six animals (3 male and 3 female) from Loro Parque zoo (Spain) were used. Faecal samples, direct behavioural observations, and clinical and training records were collected to identify meaningful relationships either between different type of potential indicators or, opportunistically, between changes in an individual’s welfare and the po- tential indicators. A methanol-based protocol was designed for the extraction of faecal cortisol metabolites, and the EIA used was successfully validated following the criteria of an immunological validation. Behavioural data sug- gested individual differences in activity budget (especially in swimming/floating ratio and affiliative behaviours). No animal suffered any remarkable health problem during this study and training records showed similar profiles between individuals. Although the current study is still collecting data, it seems that combining multiple approach- es such as behavioural, endocrinological and routine records it is a good strategy to identify and study potential welfare indicators in marine mammals under human care.

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